CARTILAGE-HOMING PEPTIDE CONJUGATES AND METHODS OF USE

THEREOF

CROSS REFERENCE

[0001] This application claims the benefit of U.S. Provisional Patent Application No.

62/472,485, filed March 16, 2017, the entire disclosure of which is incorporated by reference.

SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted

electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on March 15, 2018, is named 45639-711_601_SL.txt and is 302,571 bytes in size.

BACKGROUND

[0003] Cartilage comprises chondrocytes, a specialized cell-type which produces components of the extracellular matrix, mainly including collagen, proteoglycans (e.g., aggrecan), and elastic fibers. The extracellular matrix proteins provide support, cushion, and durability to cartilage-rich portions of the body such as joints, ears, nose, and windpipe. Cartilage is one of few tissues in the body which does not contain blood vessels and is considered an avascular tissue. Unlike many cells in the body which rely on a combination of blood flow and diffusion, chondrocytes rely on diffusion. Because it does not have a direct blood supply, compared to other connective tissues, cartilage grows and repairs much more slowly. As a result, cartilage disorders are particularly difficult to treat.

SUMMARY

[0004] The present disclosure relates to compositions and methods for treatment of cartilage disorders. Described herein are peptides that home to, migrate to, accumulate in, bind to, are retained by, or are directed to, and/or bind in cartilage following administration in a subject. In some embodiments, the homing peptides of the present disclosure are used to deliver a detection agent to image and/or diagnose cartilage, injury, or disease. In some embodiments, compositions and methods for treatment of kidney disorders are described. In other embodiments, the homing peptides of the present disclosure are used to treat or deliver an active agent to a region, tissue, structure, or cell thereof.

[0005] In some aspects, a peptide active agent conjugate comprises: a) a peptide, wherein the peptide comprises a sequence that has at least 70% sequence identity with any one of SEQ ID

NO: 24 - SEQ ID NO: 274 and upon administration to a subject the peptide homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to a cartilage of the subject, and an active agent selected from an active agent class selected from TABLE 3 or TABLE 5; b) a peptide, wherein the peptide comprises a sequence that has at least 70% sequence identity with any one of SEQ ID NO: 24 - SEQ ID NO: 274 and upon administration to a subject the peptide homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to a kidney of the subject, and an active agent selected from an active agent class selected from TABLE 4 or

TABLE 5; c) a peptide, wherein the peptide comprises a sequence that has at least 70% sequence identity with any one of SEQ ID NO: 24 - SEQ ID NO: 274 and upon administration to a subject the peptide homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to a cartilage or kidney of the subject, and an active agent selected from TABLE 3, TABLE 4, or

TABLE 5; d) a peptide, wherein the peptide comprises a sequence that has at least 70% sequence identity with any one of SEQ ID NO: 314 - SEQ ID NO: 564 and upon administration to a subject the peptide homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to a cartilage of the subject, and an active agent selected from TABLE 3 or TABLE 5; e) a peptide, wherein the peptide comprises a sequence that has at least 70% sequence identity with any one of SEQ ID NO: 314 - SEQ ID NO: 564 and upon administration to a subject the peptide homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to a kidney of the subject, and an active agent selected from an active agent class selected from TABLE 4 or

TABLE 5; f) a peptide, wherein the peptide comprises a sequence that has at least 70% sequence identity with any one of SEQ ID NO: 314 - SEQ ID NO: 564 and upon administration to a subject the peptide homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to a cartilage or kidney of the subject, and an active agent selected from an active agent class selected from TABLE 3, TABLE 4, or TABLE 5; g) a peptide, wherein the peptide comprises a sequence that has at least 70% sequence identity with any one of SEQ ID NO: 260 -

SEQ ID NO: 274 and upon administration to a subject the peptide homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to a cartilage of the subject, and an active agent selected from TABLE 3, TABLE 5, or TABLE 6; h) a peptide, wherein the peptide comprises a sequence that has at least 70% sequence identity with any one of SEQ ID NO: 260 -

SEQ ID NO: 274 and upon administration to a subject the peptide homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to a kidney of the subject, and an active agent selected from TABLE 4, TABLE 5, or TABLE 6; i) a peptide, wherein the peptide comprises a sequence that has at least 70% sequence identity with any one of SEQ ID NO: 260 -

SEQ ID NO: 274 and upon administration to a subject the peptide homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to a cartilage or a kidney of the subject, and an active agent selected from TABLE 3, TABLE 4, TABLE 5, or TABLE 6; j) a peptide, wherein the peptide comprises a sequence that has at least 70% sequence identity with any one of SEQ ID NO: 550 - SEQ ID NO: 564 and upon administration to a subject the peptide homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to a cartilage of the subject, and an active agent selected from TABLE 3, TABLE 5, or TABLE 6; k) a peptide, wherein the peptide comprises a sequence that has at least 70% sequence identity with any one of SEQ ID NO: 550- SEQ ID NO: 564 and upon administration to a subject the peptide homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to a kidney of the subject, and an active agent selected from TABLE 4, TABLE 5, or TABLE 6; or 1) a peptide, wherein the peptide comprises a sequence that has at least 70% sequence identity with any one of SEQ ID NO: 550 - SEQ ID NO: 564 and upon administration to a subject the peptide homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to a cartilage or a kidney of the subject, and an active agent selected from TABLE 3, TABLE 4, TABLE 5, or TABLE 6. In some embodiments, the peptide comprises: a) a sequence that has at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with any one of SEQ ID NO: 24 - SEQ ID NO: 274 or a fragment thereof; b) a sequence that has at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with any one of SEQ ID NO: 260 - SEQ ID NO: 274 or a fragment thereof; c) a sequence that has at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with any one of SEQ ID NO: 314 - SEQ ID NO: 564 or a fragment thereof; or d) a sequence that has at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with any one of SEQ ID NO: 550 - SEQ ID NO: 564 or a fragment thereof. In some embodiments, the peptide comprises: a) a sequence of any one of SEQ ID NO: 24 - SEQ ID NO: 274 or a fragment thereof; b) a sequence of any one of SEQ ID NO: 260 - SEQ ID NO: 274 or a fragment thereof; c) a sequence of any one of SEQ ID NO: 314 - SEQ ID NO: 564 or a fragment thereof; or d) a sequence of any one of SEQ ID NO: 550 - SEQ ID NO: 564 or a fragment thereof.

[0006] In some aspects, a peptide comprises a sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with any one of SEQ ID NO: 260- SEQ ID NO: 574 or at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with any one of SEQ ID NO: 550- SEQ ID NO: 564.

[0007] In some embodiments, the peptide comprises: a) a sequence of any one of SEQ ID NO: 1

- SEQ ID NO: 23 or a fragment thereof; b) a sequence of any one of SEQ ID NO: 275 - SEQ ID

NO: 297 or a fragment thereof; c) a sequence of any one of SEQ ID NO: 21 - SEQ ID NO: 23 or a fragment thereof; or d) a sequence of any one of SEQ ID NO: 295 - SEQ ID NO: 297 or a fragment thereof. In some embodiments, the peptide is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least, 97%, at least 98%, or at least 99% identical to any one of SEQ ID NO: 494 - SEQ ID NO: 540 or at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NO: 204 - SEQ ID NO: 250. In some embodiments, the peptide is at least 70%, at least 80%, at least 90%, at least 95%, at least 97%, or 100% identical to: a) SEQ ID NO: 111; b) SEQ ID NO: 401; c) SEQ ID NO: 24; d) SEQ ID NO: 314; e) SEQ ID NO: 27; f) SEQ ID NO: 317; g) SEQ ID NO: 185; h) SEQ ID NO: 475; i) SEQ ID NO: 30; j) SEQ ID NO: 320; k) SEQ ID NO: 108; 1) SEQ ID NO: 398; m) SEQ ID NO: 36; n) SEQ ID NO: 326; o) SEQ ID NO: 199; p) SEQ ID NO: 478; q) SEQ ID NO: 25; r) SEQ ID NO: 315; s) SEQ ID NO: 106; t) SEQ ID NO: 396; u) SEQ ID NO: 26; v) SEQ ID NO: 316; w) SEQ ID NO: 187; x) SEQ ID NO: 477; y) SEQ ID NO: 107; or z) SEQ ID NO: 397. In some embodiments, the peptide is at least 70%, at least 80%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identical to: a) SEQ ID NO: 550; b) SEQ ID NO: 551; c) SEQ ID NO: 552; d) SEQ ID NO: 553; e) SEQ ID NO: 554; f) SEQ ID NO: 555; g) SEQ ID NO: 556; h) SEQ ID NO: 557; i) SEQ ID NO: 558; j) SEQ ID NO: 559; k) SEQ ID NO: 560; 1) SEQ ID NO: 561; m) SEQ ID NO: 562; n) SEQ ID NO: 563; o) SEQ ID NO: 564; p) SEQ ID NO: 260; q) SEQ ID NO: 261; r) SEQ ID NO: 262; s) SEQ ID NO: 263; t) SEQ ID NO: 264; u) SEQ ID NO: 265; v) SEQ ID NO: 266; w) SEQ ID NO: 267; x) SEQ ID NO: 268; y) SEQ ID NO: 269; z) SEQ ID NO: 270; aa) SEQ ID NO: 271; bb) SEQ ID NO: 272; cc) SEQ ID NO: 273; or dd) SEQ ID NO: 274. In some embodiments, the peptide homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to cartilage, to kidney, or to cartilage and kidney. In some embodiments, the peptide homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to proximal tubules of the kidney. In some embodiments, the peptide is covalently conjugated to the active agent. In some embodiments, the peptide active agent conjugate homes, targets, migrates to, accumulates in, binds to, is retained by, or is directed to a cartilage or a kidney of the subject.

[0008] In some embodiments, the peptide comprises 4 or more cysteine residues. In some embodiments, the peptide comprises three or more disulfide bridges formed between cysteine residues, wherein one of the disulfide bridges passes through a loop formed by two other disulfide bridges. In some embodiments, the peptide comprises a plurality of disulfide bridges formed between cysteine residues. In some embodiments, the peptide comprises a disulfide through a disulfide knot. In some embodiments, at least one amino acid residue of the peptide is in an L configuration or, wherein at least one amino acid residue of the peptide is in a D configuration.

[0009] In some embodiments, the sequence comprises at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, at least 41, at least 42, at least 43, at least 44, at least 45, at least 46, at least 47, at least 48, at least 49, at least 50, at least 51, at least 52, at least 53, at least 54, at least 55, at least 56, at least 57, at least 58 residues, at least 59, at least 60, at least 61, at least 62, at least 63, at least 64, at least 65, at least 66, at least 67, at least 68, at least 69, at least 70, at least 71, at least 72, at least 73, at least 74, at least 75, at least 76, at least 77, at least 78, at least 79, at least 80, or at least 81 residues.

[0010] In some embodiments, any one or more K residues are replaced by an R residue or wherein any one or more R residues are replaced by for a K residue In some embodiments, any one or more M residues are replaced by any one of the I, L, or V residues. In some embodiments, any one or more L residues are replaced by any one of the V, I, or M residues. In some embodiments, any one or more I residues are replaced by any of the M, L, or V residues. In some embodiments, any one or more V residues are replaced by any of the M, I, or L residues. In some embodiments, any one or more G residues are replaced by an A residue or wherein any one or more A residues are replaced by a G residue. In some embodiments, any one or more S residues are replaced by a T residue or wherein any one or more T residues are replaced by for an S residue. In some embodiments, any one or more Q residues are replaced by an N residue or wherein any one or more N residues are replaced by a Q residue. In some embodiments, any one or more D residues are replaced by an E residue or wherein any one or more E residues are replaced by a D residue.

[0011] In some embodiments, the peptide has a charge distribution comprising an acidic region and a basic region. In some embodiments, the acidic region is a nub. In some embodiments, the basic region is a patch. In some embodiments, the peptide comprises 5-12 basic residues. In some embodiments, the peptide comprises 0-5 acidic residues. In some embodiments, the peptide comprises 6 or more basic residues and 2 or fewer acidic residues. In some embodiments, the peptide comprises a 4-19 amino acid residue fragment containing at least 2 cysteine residues, and at least 2 positively charged amino acid residues. In some embodiments, the peptide comprises a 20-70 amino acid residue fragment containing at least 2 cysteine residues, no more than 2 basic residues and at least 2 positively charged amino acid residues. In some embodiments, the peptide comprises at least 3 positively charged amino acid residues. In some embodiments, the positively charged amino acid residues are selected from K, R, or a combination thereof.

[0012] In some embodiments, the peptide has a charge greater than 2 at physiological pH. In some embodiments, the peptide has a charge greater than 3.5 at physiological pH. In some embodiments, the peptide has a charge greater than 4.5 at physiological pH. In some

embodiments, the peptide has a charge greater than 5.5 at physiological pH. In some

embodiments, the peptide has a charge greater than 6.5 at physiological pH. In some

embodiments, the peptide has a charge greater than 7.5 at physiological pH. In some

embodiments, the peptide has a charge greater than 8.5 at physiological pH. In some

embodiments, the peptide has a charge greater than 9.5 at physiological pH.

[0013] In some embodiments, the peptide is selected from a potassium channel agonist, a potassium channel antagonist, a portion of a potassium channel, a sodium channel agonist, a sodium channel antagonist, a calcium channel agonist, a calcium channel antagonist, a hadrucalcin, a theraphotoxin, a huwentoxin, a kaliotoxin, a cobatoxin, or a lectin. In some embodiments, the lectin is SHL-Ib2.

[0014] In some embodiments, the peptide is arranged in a multimeric structure with at least one other peptide.

[0015] In some embodiments, at least one residue of the peptide comprises a chemical modification. In some embodiments, the chemical modification is blocking the N-terminus of the peptide. In some embodiments, wherein the chemical modification is methylation, acetylation, or acylation. In some embodiments, the chemical modification is: methylation of one or more lysine residues or analogue thereof; methylation of the N-terminus; or methylation of one or more lysine residue or analogue thereof and methylation of the N-terminus. In some embodiments, the peptide is linked to an acyl adduct.

[0016] In some embodiments, the peptide is linked to an active agent. In some embodiments, the active agent is fused with the peptide at an N-terminus or a C-terminus of the peptide. In some embodiments, the active agent is another peptide. In some embodiments, the active agent is an antibody. In some embodiments, the active agent is an Fc domain, Fab domain, scFv, or Fv fragment. In some embodiments, the peptide fused with an Fc domain comprises a contiguous sequence. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 active agents are linked to the peptide. In some embodiments, the peptide is linked to the active agent at an N-terminus, at the epsilon amine of an internal lysine residue, at the carboxylic acid of an aspartic acid or glutamic acid residue, or a C-terminus of the peptide by a linker. In some embodiments, the peptide is linked to the active agent via a cleavable linker. In some embodiments, the peptide or peptide active agent conjugate further comprises a non-natural amino acid, wherein the non-natural amino acid is an insertion, appendage, or substitution for another amino acid.

[0017] In some embodiments, the peptide is linked to the active agent at the non-natural amino acid by a linker. In some embodiments, the linker comprises an amide bond, an ester bond, a carbamate bond, a carbonate bond, a hydrazone bond, an oxime bond, a disulfide bond, a thioester bond, a thioether bond, a triazole, a carbon-carbon bond, or a carbon-nitrogen bond. In some embodiments, the cleavable linker comprises a cleavage site for matrix metalloproteinases, thrombin, cathepsins, or beta-glucuronidase. In some embodiments, the linker is a hydro lytically labile linker. In some embodiments, the linker is pH sensitive, reducible, glutathione- sensitive, or protease cleavable. In some embodiments, the peptide is linked to the active agent via a stable linker. In some embodiments, the peptide has an isoelectric point of about 9.

[0018] In some embodiments, the peptide is linked to a detectable agent. In some embodiments, the detectable agent is fused with the peptide at an N-terminus or a C-terminus of the peptide. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 detectable agents are linked to the peptide. In some embodiments, the peptide is linked to the detectable agent via a cleavable linker. In some embodiments, the peptide is linked to the detectable agent at an N-terminus, at the epsilon amine of an internal lysine residue, or a C-terminus of the peptide by a linker. In some embodiments, the peptide active agent conjugate or peptide further comprises a non-natural amino acid, wherein the non-natural amino acid is an insertion, appendage, or substitution for another amino acid.

[0019] In some embodiments, the peptide is linked to the detectable agent at the non-natural amino acid by a linker In some embodiments, the linker comprises an amide bond, an ester bond, a carbamate bond, a hydrazone bond, an oxime bond, or a carbon-nitrogen bond. In some embodiments, the cleavable linker comprises a cleavage site for matrix metalloproteinases, thrombin, cathepsins, or beta-glucuronidase. In some embodiments, the peptide is linked to the detectable agent via a stable linker. In some embodiments, the detectable agent is a fluorophore, a near-infrared dye, a contrast agent, a nanoparticle, a metal-containing nanoparticle, a metal chelate, an X-ray contrast agent, a PET agent, a radioisotope, or a radionuclide chelator. In some embodiments, the detectable agent is a fluorescent dye.

[0020] In some aspects, a pharmaceutical composition comprises the peptide active agent conjugate of any embodiment as described herein or a salt thereof, or the peptide of any embodiment as described herein or a salt thereof, and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition is formulated for administration to a subject.

In some embodiments, the pharmaceutical composition is formulated for inhalation, intranasal administration, oral administration, topical administration, parenteral administration, intravenous administration, subcutaneous administration, intra-articular administration, intramuscular administration, intraperitoneal administration, dermal administration, transdermal administration, or a combination thereof.

[0021] In some aspects, a method of treating a condition in a subject in need thereof comprises administering to the subject the peptide active agent conjugate of any of embodiment as described herein, the peptide of any of any embodiment as described herein, or a pharmaceutical composition of any embodiment as described herein. In some embodiments, the peptide active agent conjugate, peptide, or pharmaceutical composition is administered by inhalation, intranasally, orally, topically, parenterally, intravenously, subcutaneously, intra-articularly, intramuscularly administration, intraperitoneally, dermally, transdermally, or a combination thereof. In some embodiments, the peptide active agent conjugate or the peptide homes, targets, or migrates to cartilage of the subject following administration. In some embodiments, the condition is associated with cartilage. In some embodiments, the condition is associated with a joint. In some embodiments, the condition is an inflammation, a cancer, a degradation, a growth disturbance, genetic, a tear, an infection, a disease, or an injury. In some embodiments, the condition is a chondrodystrophy. In some embodiments, the condition is a traumatic rupture or detachment. In some embodiments, the condition is a costochondritis. In some embodiments, the condition is a herniation. In some embodiments, the condition is a polychondritis. In some embodiments, the condition is a chordoma. In some embodiments, the condition is a type of arthritis. In some embodiments, the type of arthritis is rheumatoid arthritis. In some

embodiments, the type of arthritis is osteoarthritis. In some embodiments, the condition is achondroplasia. In some embodiments, the condition is benign chondroma or malignant chondrosarcoma. In some embodiments, the condition is bursitis, tendinitis, gout, pseudogout, an arthropathy, psoriatic arthritis, ankylosing spondylitis, or an infection. In some embodiments, the peptide active agent conjugate, peptide, or pharmaceutical composition is administered to treat the injury, to repair a tissue damaged by the injury, or to treat a pain caused by the injury. In some embodiments, the peptide active agent conjugate, peptide, or pharmaceutical composition is administered to treat the tear or to repair a tissue damaged by the tear. In some embodiments, the peptide active agent conjugate, peptide, or pharmaceutical composition homes, targets, or migrates to a kidney of the subject following administration. In some embodiments, the condition is associated with a kidney. In some embodiments, the condition is lupus nephritis, acute kidney injury (AKI), chronic kidney disease (CKD), hypertensive kidney damage, diabetic nephropathy, or renal fibrosis.

[0022] In some aspects, a method of imaging an organ or body region of a subject comprises: administering to the subject the peptide active agent conjugate of any embodiment as described herein, the peptide of any embodiment as described herein, or the pharmaceutical composition of any embodiment as described herein; and imaging the subject. In some embodiments, the method further comprises detecting a cancer or diseased region, tissue, structure, or cell. In some embodiments, the method further comprises performing surgery on the subject. In some embodiments, the method further comprises treating the cancer. In some embodiments, the surgery comprises removing the cancer or the diseased region, tissue, structure or cell of the subject. In some embodiments, the method further comprises imaging the cancer or diseased region, tissue, structure, or cell of the subject after surgical removal. In some embodiments, the peptide active agent conjugate is expressed as a fusion protein.

INCORPORATION BY REFERENCE

[0023] All publications, patents, and patent applications mentioned, disclosed or referenced in this specification are herein incorporated by reference in their entirety and to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE FIGURES

[0024] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings of which:

[0025] FIG. 1 illustrates the identification of the ¹⁴ C signal in the joint and other cartilage of an animal treated with the peptide of SEQ ID NO: 27.

[0026] FIG. 2 illustrates a three-dimensional structure and a line structure of a peptide of SEQ ID NO: 31.

[0027] FIG. 3 illustrates an exemplary architecture of constructs expressing sequences of SEQ ID NO: X, where X can be any one of peptides of SEQ ID NO: 24 - SEQ ID NO: 36.

[0028] FIG. 4 illustrates a schematic of a method of manufacturing of a peptide of the disclosure.

[0029] FIG. 5 illustrates alignment of SEQ ID NO: 541 (SEQ ID NO: 541 is SEQ ID NO: 27, but without the first three amino acids "GSG" and is also SEQ ID NO: 317, but without the first amino acid "G") with SEQ ID NO: 316, SEQ ID NO: 541 with SEQ ID NO: 542 (SEQ ID NO: 542 is SEQ ID NO: 30, but without the first three amino acids "GSQ" and is SEQ ID NO: 320, but without the first amino acid "Q"), and SEQ ID NO: 541 with SEQ ID NO: 483. FIG. 5A illustrates the alignment of the peptide of SEQ ID NO: 541 with the peptide of SEQ ID NO: 316. Boxes delineate conserved positively charged residues. FIG. 5B illustrates the alignment of the peptide of SEQ ID NO: 541 with the peptide of SEQ ID NO: 542. Boxes delineate conserved positively charged residues. FIG. 5C illustrates the alignment of the peptide of SEQ ID NO: 541 with the peptide of SEQ ID NO: 483. Boxes delineate conserved positively charged residues.

[0030] FIG. 6 illustrates the alignment of the peptide of SEQ ID NO: 320 with the peptide of SEQ ID NO: 484. Boxes delineate conserved positively charged residues.

[0031] FIG. 7 illustrates alignment of peptides within the pfam00451:toxin_2 structural class family of SEQ ID NO: 494 - SEQ ID NO: 540. Boxed and bolded residues indicate relative conservation of sequence while non-boxed and non-bo lded residues indicate areas of higher sequence variability.

[0032] FIG. 8 illustrates alignment of a peptide of SEQ ID NO: 494 from the pfam00451:toxin 2 structural class family with a cartilage homing peptide of this disclosure of SEQ ID NO: 27. Asterisks indicate positions with a single, fully conserved residue, a colon indicates conservation between groups of strongly similar properties (scoring > 0. 5 in the Gonnet point accepted mutation (PAM) 250 matrix), and a period indicates conservation between groups of weakly similar properties (scoring < 0. 5 in the Gonnet PAM 250 matrix).

[0033] FIG. 9 illustrates the ¹⁴ C signal in the cartilage of an animal with intact kidneys 24 hours after treatment with a peptide of SEQ ID NO: 27.

[0034] FIG. 10 shows white light images and corresponding whole body fluorescence images of a mouse administered 10 nmol of a peptide of SEQ ID NO: 108 conjugated to a Cy5. 5 fluorophore (SEQ ID NO: 108A) at 24 hours post-administration. FIG. 10A illustrates an image of a frozen section of a mouse, 24 hours after administration of 10 nmol of a peptide of SEQ ID NO: 108 conjugated to a Cy5. 5 fluorophore (SEQ ID NO: 108A). FIG. 10B illustrates the fluorescence signal in the mouse, corresponding to the section shown in FIG. 10A, 24 hours after administration of 10 nmol of a peptide of SEQ ID NO: 108 conjugated to a Cy5. 5 fluorophore (SEQ ID NO: 108A). FIG. IOC illustrates an image of a different frozen section of the mouse, 24 hours after administration of 10 nmol of a peptide of SEQ ID NO: 108 conjugated to a Cy5. 5 fluorophore (SEQ ID NO: 108A). FIG. 10D illustrates the fluorescence signal in the mouse, corresponding to the section shown in FIG. IOC, 24 hours after administration of 10 nmol of a peptide of SEQ ID NO: 108 conjugated to a Cy5. 5 fluorophore (SEQ ID NO: 108A). FIG. 10E illustrates an image of a different frozen section of the mouse, 24 hours after administration of 10 nmol of a peptide of SEQ ID NO: 108 conjugated to a Cy5. 5 fluorophore (SEQ ID NO: 108A). FIG. 10F illustrates a fluorescence signal in the mouse, corresponding to the section shown in FIG. 10E, 24 hours after administration of 10 nmol of a peptide of SEQ ID NO: 108 conjugated to a Cy5. 5 fluorophore (SEQ ID NO: 108A). [0035] FIG. 11 illustrates a multiple sequence alignment of SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO: 321, SEQ ID NO: 333, SEQ ID NO: 337, SEQ ID NO: 338, SEQ ID NO: 340, SEQ ID NO: 398, SEQ ID NO: 474, SEQ ID NO: 483, SEQ ID NO: 486, and SEQ ID NO: 543 - SEQ ID NO: 549 were used to predict enhanced peptide stability and immunogenicity. SEQ ID NO: 295 is a consensus sequence.

[0036] FIG. 12 illustrates the identification of locations the ¹⁴ C signal in the nasal, spinal, tracheal, and other cartilage of an animal treated with the peptide of SEQ ID NO: 27.

[0037] FIG. 13 shows IVIS fluorescence imaging of an isolated hind limb from a first mouse and an isolated hind limb from a second mouse after administration of 10 nmol SEQ ID NO: 108 peptide conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A). Areas of low signal intensity are shown in a thin solid line, areas of medium signal intensity are shown in a thick sold line, and areas of high signal intensity are shown in a thin dotted line. FIG. 13A shows the right hind limb with skin removed from a first mouse and from a second mouse 3 hours after peptide

administration. FIG. 13B shows the right hind limb with muscle removed from a first mouse and from a second mouse 3 hours after peptide administration of 10 nmol SEQ ID NO: 108 peptide conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A). FIG. 13C shows the right hind limb with skin removed from a first mouse and from a second mouse 24 hours after peptide administration of 10 nmol SEQ ID NO: 108 peptide conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A). FIG. 13D shows the right hind limb with muscle removed from a first mouse and from a second mouse 24 hours after peptide administration of 10 nmol SEQ ID NO: 108 peptide conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A). FIG. 13E shows the right hind limb with skin removed from a first mouse and from a second mouse 48 hours after peptide administration of 10 nmol SEQ ID NO: 108 peptide conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A). FIG. 13F shows the right hind limb with muscle removed from a first mouse and from a second mouse 48 hours after peptide administration of 10 nmol SEQ ID NO: 108 peptide conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A). FIG. 13G shows the right hind limb with skin removed from a first mouse and from a second mouse 72 hours after peptide administration of 10 nmol SEQ ID NO: 108 peptide conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A). FIG. 13H shows the right hind limb with muscle removed from a first mouse and from a second mouse 72 hours after administration of 10 nmol SEQ ID NO: 108 peptide conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A).

[0038] FIG. 14 illustrates autoradiography images of frozen sections from a mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 108. FIG. 14A illustrates the ¹⁴ C signal in a different frozen section of the mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 108. FIG. 14B illustrates the ¹⁴ C signal in a different frozen section of a mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 108

[0039] FIG. 15 illustrates autoradiography images of frozen sections from a mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 106. FIG. 15A illustrates the ¹⁴ C signal in a frozen section of a mouse. FIG. 15B illustrates the ¹⁴ C signal in a frozen section of the mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 106.

[0040] FIG. 16 illustrates autoradiography images of frozen sections from a mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 187. FIG. 16A illustrates the ¹⁴ C signal in a frozen section of the mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 187. FIG. 16B illustrates the ¹⁴ C signal in a frozen section of the mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 187.

DETAILED DESCRIPTION

[0041] The present disclosure relates generally to compositions and methods for cartilage therapy. In some embodiments, the compositions and methods herein utilize peptides that home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage following administration to a subject. In some embodiments, the cartilage homing peptides of the present disclosure exert therapeutic effect in cartilage or tissue or cell thereof. In some embodiments, the cartilage homing peptides of the present disclosure are used to deliver an active agent to cartilage or tissue or cell thereof. The active agent can exert a therapeutic effect on cartilage or tissue or cell thereof. For example, in certain embodiments, the peptide itself or the active agent allows for localized delivery of an ant i- inflammatory or other agent to cartilage or tissue or cell thereof. As another example, the active agent is a fluorophore that can be used for imaging of cartilage. In certain embodiments, the peptide itself induces therapeutic responses.

[0042] Cartilage disorders are particularly difficult to treat. A direct route for active agent administration can be parenterally (e.g., intravenously, subcutaneously, intramuscularly), intra- articular ly, by inhalation, dermally, topically, or orally. However, cartilage can be avascular thus intravenous administration of drugs can fail to reach the cartilage in significant amounts. Drugs for cartilage diseases, such as osteoarthritis, can be injected directly locally into the affected area, for example, directly injected into the joint. Few drugs aimed at treating cartilage disorders have proved therapeutically viable with lack of access to target tissue being a primary reason for failure. The lack of access to the target tissue can also lead to

administration of doses that are higher than would be necessary if a drug could home, target, or be directed to, is retained by, and/or binds to a target region, tissue, structure or cell. Thus, treatment of cartilage conditions often requires the use of high concentrations of non-specific drugs. In addition, a number of therapeutics are of interest in treating joint disorders, but are problematic because of the level of side effects caused by systemic administration of the drug (Dancevic and McCulloch, Arthritis Res Ther. 16:429 (2014)).

[0043] Specific and potent drugs that are capable of contacting the cartilage can counteract the non- specificity of many treatments by selectively targeting and delivering compounds to specific regions, tissues, cells and structures. Such drugs can also be useful to modulate ion channels, protein-protein interactions, extracellular matrix remodeling (i.e., protease inhibition), and the like. Such targeted therapy can allow for lower dosing, reduced side effects, improved patient compliance, and improvement in therapeutic outcomes, which would be advantageous not only in acute disease of the cartilage, but in chronic conditions as well.

[0044] The present disclosure provides peptides that can comprise or can be derived from cystine-dense peptides. As used herein, the term "cystine-dense peptide" can be interchangeable with the terms "knotted peptide," "knottin," and "optide," and cystine-dense peptides can also be abbreviated as "CDPs." Hitchins, amongst other disulfide-containing peptides, can also be considered "knotted peptides" or "cystine-dense peptides" for the purposes of this disclosure.

Knottins, for example, are a class of cystine-dense peptides comprising from about 11 to about

80 amino acids in length that are often folded into a compact structure. Knottins and other cystine-dense peptides are typically assembled into a complex tertiary structure that is characterized by a number of intramolecular disulfide crosslinks and can contain beta strands, an alpha helix, and other secondary structures. The presence of the disulfide bonds can give cystine-dense peptides remarkable environmental stability, allowing them to withstand extremes of temperature and pH, to resist proteolytic enzymes in the blood stream or digestive tract, and can provide specific biodistribution, pharmacokinetic, binding interactions, cellular processing, or other properties of physiologic and therapeutic value. The peptides disclosed herein can be derived from certain cystine-dense peptides. The present disclosure describes a class of cystine-dense peptides that can effectively contact cartilage and be used either directly or as carriers of active drugs, peptides, or molecules to treat a cartilage condition.

For instance, osteoarthritis is a cartilage condition that is associated with the thinning of cartilage covering the ends of bones resulting in bone directly contacting bone within the joint. Over time, the ends of the bones are subjected to increased levels of friction which ultimately causes erosion of the end of the bone. Individuals suffering from osteoarthritis experience reduced motion and increased pain. A therapeutic peptide that could contact the cartilage at the joint and ends of the bone to interact with the chondrocytes and induce increased expression of extracellular matrix proteins could be used in the treatment and prevention of osteoarthritis by increasing expression of collagen through, for example, the rate of production, amount of production, inhibition of proteins which degrade collagen, promote expression of other proteins which maintain the integrity of existing collagen proteins, or other mechanism. A peptide could also affect nearby tissues or cells such as the bone, ligaments, muscle, tendons, bursa, connective tissue, blood vessels, peripheral nerves, osteoclasts, osteoblasts, fibroblasts, synoviocytes, monocytes/macrophages, lymphocytes, plasma cells, adipocytes, endothelial cells, neurons, ligaments, muscle, tendons, and bursa.

The peptides of the disclosure can be used to treat the symptoms of various conditions. The peptides of the disclosure can bind to, home to, migrate to, accumulate in, be retained by, or be directed to cartilage and its components, including chondrocytes, extracellular matrix, collagen, hyaluranon, aggrecan (also known as cartilage- specific proteoglycan core protein

(CSPCP)), or other components of the extracellular matrix and the joint, or to other nearby components such as those described herein in joints and cartilaginous tissues as listed above.

[0045] Also described herein are peptides that selectively home, target, are directed to, migrate to, are retained by, or accumulate in and/or bind to specific regions, tissues, structures or cells of the cartilage that aid in managing, decreasing, ablating or reducing pain (e.g., joint pain) due to chronic disease or cartilage injury or other therapeutic indications as described herein. A peptide that homes, targets, migrates to, is directed to, is retained by, or accumulates in and/or binds to one or more specific regions, tissues, structures or cells of the cartilage can have fewer off-target and potentially negative effects, for example, side effects that often limit use and efficacy of pain drugs. In addition, such peptides can reduce dosage and increase the efficacy of existing drugs by directly targeting them to a specific region, tissue, structure or cell of the cartilage and helping the contact the cartilage or increasing the local concentration of agent. The peptide itself can modulate pain or it can be conjugated to an agent that modulates pain. Such pain modulation may operate by various mechanisms such as

modulating inflammation, autoimmune responses, direct or indirect action on pain receptors, cell killing, or programmed cell death (whether via an apoptotic and/or non-apoptotic pathway of diseased cells or tissues, and the like (Tait et al., Cell Sci 127(Pt 10):2135-44 (2014)).

[0046] Peptides of this disclosure that home, target, are directed to, migrate to, are retained by, accumulate in, or bind to specific regions, tissues, structures or cells of the cartilage can do so with different degrees of efficiency. Peptides can have a higher concentration in cartilage than in other locations, such as blood or muscle. Peptides can be recorded as having a signal in cartilage as a percentage of signal in blood. For example, a cartilage signal of 200% indicates that the signal in cartilage is twice as high as the signal in blood. In some embodiments, peptides that have cartilage homing properties can have a cartilage signal of >170% by radiographic densitometry measurements. In other embodiments, peptides that are cartilage homers can have a cartilage signal of >200% by radiographic densitometry measurements. In other embodiments, peptides that are more efficient cartilage homers can have a cartilage signal of >300% by radiographic densitometry measurements. In other embodiments, peptides that are more efficient cartilage homers can have a cartilage signal of >400% by radiographic densitometry measurements. In other embodiments, peptides that are strongest cartilage homers of highest interest can have a cartilage signal of >500% by radiographic densitometry measurements. In some embodiments, measurement of the ratio of peptide concentration in blood, muscle, or other tissues relative to the peptide concentration in cartilage can be performed using various methods including measuring the densitometry signal of peptides labeled with radioisotopes (as described above), or by using other assays.

[0047] Peptides that selectively home, target, are directed to, migrate to, are retained by, or accumulate in and/or bind to specific regions, tissues, structures or cells of the cartilage can occur after administration of the peptide to a subject. A subject can be a human or a non- human animal.

[0048] The peptides disclosed herein can be used as active agents, or conjugated to detection agents such a fluorophores, iodide-containing X-ray contrast agents, lanthanide chelates (e.g., gadolinium for MRI imaging), perfluorocarbons (for ultrasound), or PET tracers (e.g., 18F or

11C) for imaging and tracing the peptide, or conjugated to agents such as ant i- inflammatory active agents or other active agents to the joint to treat inflammation or other disease.

[0049] The peptides disclosed herein can be used to bind cartilage explants ex vivo. Cartilage explants can be from any subject, such as a human or an animal. Assessment of peptide binding to cartilage explants can be used to screen peptides that may efficiently home to cartilage in vivo

[0050] In some embodiments, peptides of this disclosure home, target, are directed to, migrate to, are retained by, accumulate in, or bind to specific regions, tissues, structures or cells of the kidneys. For example, in some embodiments, peptides of this disclosure home, target, are directed to, migrate to, are retained by, accumulate in, or bind to the proximal tubules of the kidneys, kidney nephrons, or podocytes. Peptides that selectively home, target, are directed to, migrate to, are retained by, or accumulate in and/or bind to specific regions, tissues, structures or cells of the kidney can occur after administration of the peptide to a subject. A subject can be a human or a non-human animal. The peptides disclosed herein can be used as active agents, or conjugated to detection agents such a fluorophores, iodide-containing X-ray contrast agents, lanthanide chelates (e.g., gadolinium for MRI imaging), perfluorocarbons (for ultrasound), or PET tracers (e.g., 18F or 11C) for imaging and tracing the peptide, or conjugated to agents such as anti-inflammatory agents or other agents to the kidney to treat renal cancer, chronic kidney failure or other kidney disease.

[0051] One roadblock in the advancement and wide spread use of peptides as a therapeutic is that peptides can be chemically and physically unstable. During the process of manufacturing of therapeutic peptides essential considerations can include storage conditions, sustained

biochemical function, and in vivo delivery. Peptide degradation products can result in the formation of species that alter the safety profile, potency, and immunogenicity of the peptide. These peptide degradation products can form during manufacture and storage, as well as in vivo after delivery to a patient. Furthermore, peptide degradation may limit the shelf- life and increase production cost due to unstable peptides requiring refrigeration or shipment on dry ice. The latter can necessitate continual monitoring and validation of peptides as degradation products could have formed during the manufacturing process. Hence, there is an urgent need for the rationale design and production of therapeutic peptides that have enhanced stability, for example, in the ambient environment, during the process of manufacturing, in storage, and that prevent the likelihood of peptide degradation under a variety of conditions.

[0052] In some embodiments, the peptides and peptide-drug conjugates of the present disclosure have stability properties that minimize peptide or peptide-drug conjugate degradation to enable adequate storage. Long term, accelerated, and intermediate storage conditions for the peptides and peptide-drug conjugates of the present disclosure can include long term storage conditions of

25°C + 2°C/60% relative humidity (RH) + 5% RH, or 30°C + 2°C/65% RH + 5% RH for at least

6 months, at least 12 months, and up to 1 year, up to 2 years, up to 3 years, up to 4 years, or longer than 4 years. In addition, intermediate and short term storage conditions (e.g., during transport, distribution, manufacturing, or handling), or long term storage conditions for certain climates and infrastructures, can include storage conditions of 30°C + 2°C/65% RH + 5% RH or

40°C + 2°C/75% RH + 5% RH for up to 1 hour, for up to 8 hours, for up to 1 day, for up to 3 days, for up to 1 week, for up to 1 month, for up to 3 months, for up to 6 months or at least 6 months, up to 1 year, up to 2 years, up to 3 years, up to 4 years, or longer than 4 years).

Moreover, the peptides and peptide-drug conjugates of the present disclosure can be refrigerated, for example between 5°C + 3°C for at least 6 months, at least 12 months, and up to 1 year, up to

2 years, up to 3 years, up to 4 years, or longer than 4 years. In addition, intermediate and short term refrigeration conditions (e.g., during transport, distribution, manufacturing, or handling) can include 25°C + 2°C/60% RH + 5% RH for up to 1 hour, for up to 8 hours, for up to 1 day, for up to 3 days, for up to 1 week, for up to 1 month, for up to 3 months, for up to 6 months or at least 6 months, and potentially longer (at least 12 months and up to 1 year, up to 2 years, up to 3 years, up to 4 years, or longer than 4 years). Such conditions for storage, whether based on ambient or refrigerated conditions can be adjusted based upon the four zones in the world (e.g., the

International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) stability Zone I, II, III, or IV) that are distinguished by their characteristic prevalent annual climatic conditions. In addition, formulation components can be principally chosen for their ability to preserve the native conformation and chemical structure of the peptides and peptide-drug conjugates of the present disclosure in storage by preventing denaturation due to hydrophobic interactions and aggregation, as well as by preventing chemical degradation, including truncation, oxidation, deamidation, cleavage, hydrolysis, isomerization, disulfide exchange, racemization, and beta elimination (Cleland, et al., Crit Rev Ther Drug Carrier Syst 10(4): 307-377 (1993); Shire et al., J Pharm Sci 93(6): 1390-1402 (2004); Wakankar and

Borchardt, J Pharm Sci 95(11): 2321-2336 (2006)).

[0053] In some embodiments, the peptides and peptide-drug conjugates of the present disclosure have incorporated properties that minimize immunogenicity of the peptides and peptide-drug conjugates. Immunogenicity can be a major concern with the development of therapeutic peptides and proteins, and there is an urgent need for the rationale design and production of therapeutic peptides that have reduced immunogenicity and that increase their safety and efficacy. Immunogenicity can occur against a desired peptide sequence or a peptide degradation product. Immunogenicity can occur when a patient develops an immune response to the therapeutic peptide, protein, conjugate, or other drug, such as by producing antibodies that bind to and/or neutralize the therapeutic peptide, protein, conjugate, or other drug. The likelihood of immunogenicity can increase when drugs are administered more than once or chronically.

Immunogenicity can reduce patient exposure to the drug, can reduce effectiveness of the drug, and can also result in safety risks for the patient, such as generating an immune response to self- proteins or other adverse responses related to increased immunogenicity to the therapeutic peptide, protein, conjugate, or other drug. Immunogenic responses can vary from patient to patient and also amongst different groups of HLA alleles, as well as over time. As such, minimizing risk of immunogenicity with a therapeutic peptide or protein can be important for developing a drug that can be effectively and safely used for treatment. Various methods exist for assessment of immunogenic potential, which can include in silico methods, in vitro testing, preclinical in vivo testing, and assessment during clinical dosing. Evaluation early in product design and development of the therapeutic peptides and peptide-drug conjugates of the present disclosure in the in vivo milieu in which they function (e.g., in inflammatory environments or at physiologic pH) can reveal susceptibilities to modifications (e.g., aggregation and deamidation) that can result in loss of efficacy or induction of immune responses. Such information can be used to facilitate product engineering to enhance the stability of the product under such in vivo conditions or reduce immunogenicity. Moreover, the therapeutic peptides and peptide-drug conjugates of the present disclosure can be designed to minimize protein aggregation. Strategies to minimize aggregate formation can be used early in drug development, for example, by using an appropriate cell substrate, selecting manufacturing conditions that minimize aggregate formation, employing a robust purification scheme that removes aggregates to the greatest extent possible, and choosing a formulation and container closure system that minimize aggregation during storage.

[0054] Additional aspects and advantages of the present disclosure will become apparent to those skilled in this art from the following detailed description, wherein illustrative embodiments of the present disclosure are shown and described. As will be realized, the present disclosure is capable of other and different embodiments, and its several details are capable of modifications in various respects, all without departing from the disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature, and not as restrictive.

[0055] As used herein, the abbreviations for the natural L- enantiomeric amino acids are conventional and are as follows: alanine (A, Ala); arginine (R, Arg); asparagine (N, Asn);

aspartic acid (D, Asp); cysteine (C, Cys); glutamic acid (E, Glu); glutamine (Q, Gin); glycine (G, Gly); histidine (H, His); isoleucine (I, He); leucine (L, Leu); lysine (K, Lys); methionine (M, Met); phenylalanine (F, Phe); proline (P, Pro); serine (S, Ser); threonine (T, Thr); tryptophan (W, Trp); tyrosine (Y, Tyr); valine (V, Val). Typically, Xaa can indicate any amino acid. In some embodiments, X can be asparagine (N), glutamine (Q), histidine (H), lysine (K), or arginine (R).

[0056] Some embodiments of the disclosure contemplate D-amino acid residues of any standard or non-standard amino acid or analogue thereof. When an amino acid sequence is represented as a series of three-letter or one-letter amino acid abbreviations, the left-hand direction is the amino terminal direction and the right-hand direction is the carboxyl terminal direction, in accordance with standard usage and convention.

Peptides

The cystine-dense peptides herein can bind targets with antibody- like affinity. The cystine-dense peptides can modulate the activity of a plurality of cartilage regions, tissues, structures or cells. For example, in some embodiments, the cystine-dense peptide conjugated to a bone-modifying drug homes to the cartilage of a diseased joint and releases the drug, creating a higher local concentration of drug in an area of eroded or damaged bone than would be achieved without the cartilage targeting function of the peptide. The cystine-dense peptide can be conjugated to a drug that can affect nearby tissues or cells such as the ligaments, muscle, tendons, bursa, connective tissue, blood vessels, peripheral nerves, osteoclasts, osteoblasts, fibroblasts, synoviocytes, monocytes/macrophages, lymphocytes, plasma cells, adipocytes, endothelial cells, neurons, ligaments, muscle, tendons, and bursa. The cystine-dense peptide conjugated to a drug can bind to, home to, migrate to, accumulate in, be retained by, or be directed to cartilage and its components, including chondrocytes, extracellular matrix, collagen of any type, hyaluranon, aggrecan (also known as cartilage- specific proteoglycan core protein (CSPCP)), proteoglycans, glycoasminoglycans, glycoproteins, decorin, biclycan, fibromodulin, or other components of the extracellular matrix and the joint, or to other nearby components such as those described herein in joints and cartilaginous tissues as listed above. Some of the cartilage regions, tissues, and structures that peptides and peptide-drug conjugates can target to treat a cartilage-associated disorder include: (a) elastic cartilage; (b) hyaline cartilage, such as articular cartilage and physeal cartilage; (c) fibrocartilage; and (d) any cells or cell types in (a) -(c) above. Some of the areas where the peptide and peptide-drug conjugates can target to treat a cartilage- associated disorder include: cartilage includes joints such as knees, hips, or digits, nasal cartilage, spinal cartilage, tracheal cartilage, and rib cartilage. In various aspects, cartilage components include aggrecan and type II collagen. Additionally, in some embodiments, cystine-dense peptides can penetrate into cells. In other embodiments, cystine-dense peptides do not enter cells. In other embodiments, cystine-dense peptides exhibit more rapid clearance and cellular uptake compared to other types of molecules.

[0057] The peptides of the present disclosure can comprise cysteine amino acid residues. In some cases, the peptide has at least 4 cysteine amino acid residues. In some cases, the peptide has at least 6 cysteine amino acid residues. In other cases, the peptide has at least 8 cysteine amino acid residues, at least 10 cysteine amino acid residues, at least 12 cysteine amino acid residues, at least 14 cysteine amino acid residues or at least 16 cysteine amino acid residues.

[0058] A cystine-dense peptide can comprise disulfide bridges. A cystine-dense can be a peptide wherein 5% or more of the residues are cysteines forming intramolecular disulfide bonds as cystines. A disulfide-linked peptide can be a drug scaffold. In some embodiments, the disulfide bridges form an inhibitor knot. A disulfide bridge can be formed between cysteine residues, for example, between cysteines 1 and 4, 2 and 5, or, 3 and 6. In some cases, one disulfide bridge passes through a loop formed by the other two disulfide bridges, for example, to form the inhibitor knot. In other cases, the disulfide bridges can be formed between any two cysteine residues.

[0059] The present disclosure further includes peptide scaffolds that, e.g., can be used as a starting point for generating additional peptides that can target and home to cartilage. In some embodiments, these scaffolds can be derived from a variety of cystine-dense peptides. In certain embodiments, cystine-dense peptides are assembled into a complex tertiary structure that is characterized by a number of intramolecular disulfide crosslinks, and optionally contain beta strands and other secondary structures such as an alpha helix. For example, cystine-dense peptides include, in some embodiments, small disulfide-rich proteins characterized by a disulfide through disulfide knot. This knot can be, e.g., obtained when one disulfide bridge crosses the macrocycle formed by two other disulfides and the interconnecting backbone. In some embodiments, the cystine-dense peptides can include growth factor cysteine knots or inhibitor cysteine knots. Other possible peptide structures can include peptide having two parallel helices linked by two disulfide bridges without β- sheets (e.g., hefutoxin).

[0060] A cystine-dense peptide can comprise at least one amino acid residue in an L

configuration. A cystine-dense peptide can comprise at least one amino acid residue in a D configuration. In some embodiments, a cystine-dense peptide is 15-40 amino acid residues long. In other embodiments, a cystine-dense peptide is 11-57 amino acid residues long. In further embodiments, a cystine-dense peptide is at least 20 amino acid residues long.

[0061] In some embodiments, the peptides are members of the pfam00451:toxin_2 family. The pfam00451:toxin_2 structural class family can include a peptide of any one of SEQ ID NO: 494 - SEQ ID NO: 540. A cartilage homing peptide of this disclosure can be a variant of any peptide members of the pfam00451:toxin_2 family. In some embodiments, an exemplary cartilage homing peptide of this disclosure that is a variant of the pfam00451:toxin_2 structural class family is a peptide of SEQ ID NO: 27. In other embodiments, an exemplary cartilage homing peptide of this disclosure that is a variant of the pfam00451:toxin_2 structural class family is a peptide of SEQ ID NO: 108. In other embodiments, the variant peptides are at least 30% identical to a peptide of the structural class pfam00451:toxin_2 family. In some embodiments, the variant peptides are 30%, 40%, 50%, 60%, 80%, 90% or 95% identical to a peptide of the structural class pfam00451:toxin_2 family. In some embodiments, the variant peptides are at least 30%, at least 40%, at least 50%, at least 60%, at least 80%, at least 90% or at least 95% identical to a peptide of the structural class pfam00451:toxin_2 family.

[0062] In some embodiments, cartilage homing peptides are family members the sequences

GSXVXXXVKCXGSKQCXXPCKRXXGXRXGKCINKKXCKCYXXX (SEQ ID NO: 9) or

XVXXXVKCXGSKQCXXPCKRXXGXRXGKCINKKXCKCYXXX (SEQ ID NO: 283) wherein X can be any amino acid or amino acid analogue or null, in which these sequences are based on the most common elements found in the following sequences:

GSGVPINVKCRGSRDCLDPCKKA-GMRFGKCINSK-CHCTP— (SEQ ID NO: 27),

GS-VRIPVSCKHSGQCLKPCKDA-GMRFGKCMNGK-CDCTPK- (SEQ ID NO: 26),

GSQVQTNVKCQGGS-CASVCRREIGVAAGKCINGK-CVCYRN- (SEQ ID NO: 30),

GS— -ISCTGSKQCYDPCKRKTGCPNAKCMNKS-CKCYGCG (SEQ ID NO: 29), GSEV— IRCSGSKQCYGPCKQQTGCTNSKCMNKV-CKCYGCG (SEQ ID NO: 31),

GSAVCVYRT CDKDCKRR-GYRSGKCINNA-CKCYPYG (SEQ ID NO: 28),

GS— GI VC- - - KVCKIICGMQ- GKKVNIC KAPIKC KC KKG- (SEQ ID NO: 24), and

GS QIYTS KECNGS SEC YSHCEGITGKRS GKCINKK-C YC YR— (SEQ ID NO: 33), where the following residues may be independently interchanged in the sequences: K and R; M, I, L, and V; G and A; S and T; Q and N; and X can independently be any number of any amino acid or no amino acid. The N-terminal GS sequence can be included or excluded between the peptides of the present disclosure.

[0063] In some embodiments, cartilage homing peptides are family members of the sequences

GSXVXIXVKCXGSXQCLXPCKXAXGXRXGKCMNGKCXCXPXX (SEQ ID NO. 21) or

XVXIXVKCXGSXQCLXPCKXAXGXRXGKCMNGKCXCXPXX (SEQ ID NO. 295), wherein X can be any amino acid or amino acid analogue or null, in which these sequences are based on the most common elements found in the following sequences:

-VRIPVSCKHSGQCLKPCKDA-GMRFGKCMNGKCDCTPK- (SEQ ID NO: 316),

GVPINVKCRGSRDCLDPCKKA-GMRFGKCINSKCHCTP— (SEQ ID NO: 317),

- - -E VIRCS GS KQC YGPC KQQTGCTNS KCMNKVC KC YGC G (SEQ ID NO: 321),

G VIIN VKC KIS RQCLEPC KKA- GMRFGKCMNGKCHCTPK- (SEQ ID NO: 333),

GVPTDVKCRGSPQCIQPCKDA-GMRFGKCMNGKCHCTPK- (SEQ ID NO: 337),

GVPINVSCTGSPQCIKPCKDA-GMRFGKCMNRKCHCTPK- (SEQ ID NO: 338),

-VGINVKCKHSGQCLKPCKDA-GMRFGKCINGKCDCTPK- (SEQ ID NO: 340),

G VPIN VRCRGS RDCLDPCRR A- GMRFGRCINS RCHCTP- - (SEQ ID NO: 398),

QKILS NRCNNS S ECIPHCIRIFGTR A AKCINRKC YC YP- - (SEQ ID NO: 474),

-VFINVKCRGSPECLPKCKEAIGKSAGKCMNGKCKCYP— (SEQ ID NO: 483),

-VPTDVKCRGSPQCIQPCKDA-GMRFGKCMNGKCHCTP- (SEQ ID NO: 486),

— AEIIRCSGTRECYAPCQKLTGCLNAKCMNKACKCYGCV (SEQ ID NO: 543),

-RPTDIKCSASYQCFPVCKSRFGKTNGRCVNGLCDCF— (SEQ ID NO: 544),

-QFTDVKCTGSKQCWPVCKQMFGKPNGKCMNGKCRCYS-- (SEQ ID NO: 545),

-VGINVKCKHSRQCLKPCKDA-GMRFGKCTNGKCHCTPK- (SEQ ID NO: 546),

-VVIGQRCYRSPDCYSACKKLVGKATGKCTNGRCDC— - (SEQ ID NO: 547),

— NFKVEGACSKPCRKYCIDK-GARNGKCINGRCHCYY— (SEQ ID NO: 548), and

QIDTNVKCS GS S KC VKICIDRYNTRG AKCINGRCTC YP-- (SEQ ID NO: 549).

[0064] In some embodiments, the cartilage homing peptides are family members of the sequences GSXVXIXVRCXGSXQCLXPCRXAXGXRXGRCMNGRCXCXPXX (SEQ ID NO:

22) or XVXIXVRCXGSXQCLXPCRXAXGXRXGRCMNGRCXCXPXX (SEQ ID NO: 296) wherein X can be any amino acid or amino acid analogue or null, in which these sequences are based on the most common elements found in the following sequences and with K interchanged with R: SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO: 321, SEQ ID NO: 333, SEQ ID NO: 337,SEQ ID NO: 338, SEQ ID NO: 340, SEQ ID NO: 398, SEQ ID NO: 483, SEQ ID NO: 486, or SEQ ID NO: 543 - SEQ ID NO: 549.

[0065] In some embodiments, a peptide comprises the sequence

GSGVPIX RCRGSRDCX PCRRAGX^FGRCIX^RCX^X ⁸ ? (SEQ ID NO: 23) or GVPIXVRCRGSRDCX PCRRAGX^FGRCIX^RCX^X ⁸ ? (SEQ ID NO: 297), where the following residues where X 1 is selected from N, S, or G, wherein X 2 is selected from L or Y, wherein X 3 is selected from D or E, wherein X 4 is selected from M or T, wherein X 5 is selected from N, Q, A, S, T, or L, wherein X ⁶ is selected from S, G, or R, wherein X ⁷ is selected from H or Y, and wherein X is selected from T or Y. In some embodiments, zero or one or more of the R residues in SEQ ID NO: 23 or SEQ ID NO: 297 can be replaced with K residues. In some embodiments, zero or one or more of the R residues in SEQ ID NO: 23 or SEQ ID NO: 297 can be replaced with A residues. In other embodiments, zero or one or more R residues in SEQ ID NO: 23 or SEQ ID NO: 297 can each be replaced with either a K or an A residue in any combination. In other embodiments, peptides are family members of the sequence

GSXXXGCVXXXXKCRPGXKXCCXPXKRCSRRFGXXXXKKCKXXXXXX (SEQ ID NO: 10) or XXXGCVXXXXKCRPGXKXCCXPXKRCSRRFGXXXXKKCKXXXXXX (SEQ ID NO: 284), in which the sequence is based on the most common elements found in the following sequences:

GS— ACKGVFDACTPGKNECC-PNRVCSDK-H— KWCKWKL— (SEQ ID NO: 32),

GS— GCLEFWWKCNPNDD KCCRPKLKCS KLF KLCNFS FG- - (SEQ ID NO: 34),

GSSEKDCIKHLQRCR-ENKDCC--SKKCSRR-GTNPEKRCR (SEQ ID NO: 25), and

GS— GCFGY— KCDYY-KGCCSGYV-CSPTW KWCVRPGPGR (SEQ ID NO: 36), where the following residues may be independently interchanged in the sequences: K and R; M, I, L, and V; G and A; S and T; Q and N; and X can independently be any number of any amino acid or no amino acid. The N-terminal GS sequence can be included or excluded between the peptides of the present disclosure.

[0066] In some embodiments, a peptide comprises the sequence

GSGVX ¹ IX ² X ³ KCX ⁴ GSKQCX ⁵ DPCKX ⁶ X ⁷ X ⁸ GX ⁹ RX ¹⁰ GKCX ¹¹ NKKCKCX ¹² X ¹³ X ¹⁴ X ¹⁵ (SEQ ID NO: 1) or

GVX ¹ IX ² X ³ KCX ⁴ GSKQCX ⁵ DPCKX ⁶ X ⁷ X ⁸ GX ⁹ RX ¹⁰ GKCX ¹¹ NKKCKCX ¹² X ¹³ X ¹⁴ X ¹⁵ (SEQ ID NO: 275), wherein X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ , X ⁹ , X ¹⁰ , X ¹¹ , X ¹² , X ¹³ , X ¹⁴ and X ¹⁵ are each individually any amino acid or amino acid analogue or null. In some cases, the peptide comprises the sequence GSGVX ¹ IX ² X ³ KCX ⁴ GSKQCX ⁵ DPCKX ⁶ X ⁷ X ⁸ GX ⁹ RX ¹⁰ GKCX ¹¹ NKKCKCX ¹² X ¹³ X ¹⁴ X ¹⁵ (SEQ ID NO: 2) or

GVX ¹ IX ² X ³ KCX ⁴ GSKQCX ⁵ DPCKX ⁶ X ⁷ X ⁸ GX ⁹ RX ¹⁰ GKCX ¹¹ NKKCKCX ¹² X ¹³ X ¹⁴ X ¹⁵ (SEQ ID

NO: 276), where X 1 is selected from P or R, wherein X2 is selected from P or N, wherein X 3 is selected from V or I, wherein X ⁴ is selected from S, T, R or K, wherein X ⁵ is selected from Y or

L, wherein X 6 is selected from Q, R or K, wherein X 7 is selected from A, K or R, wherein X 8 is selected from T or A, wherein X ⁹ is selected from C or M, wherein X ¹⁰ is selected from F or N, wherein X 11 is selected from M or I, wherein X 12 is selected from Y or T, wherein X 13 is selected from G or P, wherein X ¹⁴ is selected from C or null, and wherein X ¹⁵ is selected from G or null.

[0067] In some embodiments, a peptide comprises the sequence

GSX ¹ X ² X ³ X ⁴ IX ⁵ CX ⁶ GSKQCYX ⁷ PCKX ⁸ X ⁹ TGCX ¹⁰ X ¹¹ X ¹² KCX ¹³ X ¹⁴ KX ¹⁵ CKCYGCG (SEQ ID NO: 3) or X ¹ X ² X ³ X ⁴ IX ⁵ CX ⁶ GSKQCYX ⁷ PCKX ⁸ X ⁹ TGCX ¹⁰ X ¹¹ X ¹² KCX ¹³ X ¹⁴ KX ¹⁵ CKCYGCG (SEQ ID NO: 277), wherein X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ , X ⁹ , X ¹⁰ , X ¹¹ , X ¹² , X ¹³ , X ¹⁴ , and X ¹⁵ are each individually any amino acid or amino acid analogue or null. In some cases, the peptide comprises the sequence

GSX ¹ X ² X ³ X ⁴ IX ⁵ CX ⁶ GSKQCYX ⁷ PCKX ⁸ X ⁹ TGCX ¹⁰ X ¹¹ X ¹² KCX ¹³ X ¹⁴ KX ¹⁵ CKCYGCG (SEQ ID NO: 4) or X ¹ X ² X ³ X ⁴ IX ⁵ CX ⁶ GSKQCYX ⁷ PCKX ⁸ X ⁹ TGCX ¹⁰ X ¹¹ X ¹² KCX ¹³ X ¹⁴ KX ¹⁵ CKCYGCG

(SEQ ID NO: 278), where X 1 is selected from G or null, wherein X 2 is selected from S or null, wherein X 3 is selected from E, G or null, wherein X 4 is selected from V, S, or null, wherein X 5 is selected from R or S, wherein X ⁶ is selected from S or T, wherein X ⁷ is selected from G or D, wherein X ⁸ is selected from Q or R, wherein X ⁹ is selected from Q or K, wherein X ¹⁰ is selected from T or P, wherein X 11 is selected from N or Q, wherein X 12 is selected from S or A, wherein X ¹³ is selected from M or L, wherein X ¹⁴ is selected from N or Q, and wherein X ¹⁵ is selected from V or S.

[0068] In some embodiments, a peptide comprises the sequence

GSX ¹ X ² X ³ VX ⁴ IX ⁵ VX ⁶ CX ⁷ X ⁸ SX ⁹ X ¹⁰ CLX ¹¹ PCKX ¹² AGMRFGKCX ¹³ NX ¹⁴ KCX ¹⁵ CTPX ¹⁶ (SEQ ID NO: 5) or

χ ^! χ ² χ ³ VX ⁴ IX ⁵ VX ⁶ CX ⁷ X ⁸ SX ⁹ X ¹⁰ CLX ¹ ^CKX^AGMRFGKCX^NX^KCX^CTPX ¹⁶ (SEQ ID NO: 279), wherein X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ , X ⁹ , X ¹⁰ , X ¹¹ , X ¹² , X ¹³ , X ¹⁴ , X ¹⁵ , X ¹⁶ are each individually any amino acid or amino acid analogue or null. In some cases, the peptide comprises the sequence

GSX ¹ X ² X ³ VX ⁴ IX ⁵ VX ⁶ CX ⁷ X ⁸ SX ⁹ X ¹⁰ CLX ¹¹ PCKX ¹² AGMRFGKCX ¹³ NX ¹⁴ KCX ¹⁵ CTPX ¹⁶ (SEQ ID NO: 6) or

χ ^! χ ² χ ³ VX ⁴ IX ⁵ VX ⁶ CX ⁷ X ⁸ SX ⁹ X ¹⁰ CLX ¹ ^CKX^AGMRFGKCX^NX^KCX^CTPX ¹⁶ (SEQ ID

NO: 280), where X 1 is selected from G or null, wherein X 2 is selected from G, S or null, wherein X 3 is selected from G, S or null, wherein X 4 is selected from P or R, wherein X 5 is selected from

N or P, wherein X 6 is selected from K or S, wherein X 7 is selected from R or K, wherein X 8 is selected from G or H, wherein X ⁹ is selected from R or G, wherein X ¹⁰ is selected from D or Q, wherein X 11 is selected from D or K, wherein X 12 is selected from K or D, wherein X 13 is selected from I or M, wherein X ¹⁴ is selected from S or G, wherein X ¹⁵ is selected from H or D, and wherein X ¹⁶ is selected from K or null.

[0069] In some embodiments, a peptide comprises the sequence

GSXVXVKCXGSKQCXPCKRXGXRXGKCINKKXCKCYX (SEQ ID NO: 7) or

GSXGCVXKCRPGXKXCCXPXKRCSRRFGXKKCKX (SEQ ID NO: 8), wherein each letter is each individually any amino acid or amino acid analogue and where X is no amino acid or a 1-10 amino acid long peptide fragment wherein each amino acid within such peptide fragment can in each case be any amino acid or amino acid analogue. In some embodiments, a peptide comprises the sequence XVXVKCXGSKQCXPCKRXGXRXGKCINKKXCKCYX (SEQ ID NO: 281) or XGCVXKCRPGXKXCCXPXKRCSRRFGXKKCKX (SEQ ID NO: 282), wherein each letter is each individually any amino acid or amino acid analogue and where X is no amino acid or a 1-10 amino acid long peptide fragment wherein each amino acid within such peptide fragment can in each case be any amino acid or amino acid analogue.

[0070] In some embodiments, a peptide comprises the sequence

GSGVX ¹ IX ² X ³ RCX ⁴ GSRQCX ⁵ DPCRX ⁶ X ⁷ X ⁸ GX ⁹ RX ¹⁰ GRCX ¹¹ NRRCRCX ¹² X ¹³ X ¹⁴ X ¹⁵ (SEQ ID NO: 11) or

GVX ¹ IX ² X ³ RCX ⁴ GSRQCX ⁵ DPCRX ⁶ X ⁷ X ⁸ GX ⁹ RX ¹⁰ GRCX ¹¹ NRRCRCX ¹² X ¹³ X ¹⁴ X ¹⁵ (SEQ ID NO: 285), wherein X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ , X ⁹ , X ¹⁰ , X ¹¹ , X ¹² , X ¹³ , X ¹⁴ , and X ¹⁵ are each individually any amino acid or amino acid analogue or null. In some cases, the peptide comprises the sequence

GSGVX ¹ IX ² X ³ RCX ⁴ GSRQCX ⁵ DPCRX ⁶ X ⁷ X ⁸ GX ⁹ RX ¹⁰ GRCX ¹¹ NRRCRCX ¹² X ¹³ X ¹⁴ X ¹⁵ (SEQ ID NO: 12) or

GVX ¹ IX ² X ³ RCX ⁴ GSRQCX ⁵ DPCRX ⁶ X ⁷ X ⁸ GX ⁹ RX ¹⁰ GRCX ¹ ^RRCRCX^W ⁵ (SEQ ID

NO: 286), where X 1 is selected from P or R, wherein X2 is selected from P or N, wherein X 3 is selected from V or I, wherein X ⁴ is selected from S, T, R or K, wherein X ⁵ is selected from Y or

L, wherein X 6 is selected from Q, R or K, wherein X 7 is selected from A, K or R, wherein X 8 is selected from T or A, wherein X ⁹ is selected from C or M, wherein X ¹⁰ is selected from F or N, wherein X 11 is selected from M or I, wherein X 12 is selected from Y or T, wherein X 13 is selected from G or P, wherein X ¹⁴ is selected from C or null, and wherein X ¹⁵ is selected from G or null.

[0071] In some embodiments, a peptide comprises the sequence

GSX ¹ X ² X ³ X ⁴ IX ⁵ CX ⁶ GSRQCYX ⁷ PCRX ⁸ X ⁹ TGCX ¹⁰ X ¹ ^^RCX^X^RX^CRCYGCG (SEQ ID NO: 13) or X ¹ X ² X ³ X ⁴ IX ⁵ CX ⁶ GSRQCYX ⁷ PCRX ⁸ X ⁹ TGCX ¹⁰ X ¹¹ X ¹² RCX ¹³ X ¹⁴ RX ¹⁵ CRCYGCG (SEQ ID NO: 287), wherein X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ , X ⁹ , X ¹⁰ , X ¹¹ , X ¹² , X ¹³ , X ¹⁴ , and X ¹⁵ are each individually any amino acid or amino acid analogue or null. In some cases, the peptide comprises the sequence

GSX ¹ X ² X ³ X ⁴ IX ⁵ CX ⁶ GSRQCYX ⁷ PCRX ⁸ X ⁹ TGCX ¹⁰ X ¹ ^^RCX^X^RX^CRCYGCG, (SEQ ID NO: 14) or X ¹ X ² X ³ X ⁴ IX ⁵ CX ⁶ GSRQCYX ⁷ PCRX ⁸ X ⁹ TGCX ¹⁰ X ¹¹ X ¹² RCX ¹³ X ¹⁴ RX ¹⁵ CRCYGCG

(SEQ ID NO: 288), where X 1 is selected from G or null, wherein X 2 is selected from S or null, wherein X 3 is selected from E, G or null, wherein X 4 is selected from V, S, or null, wherein X 5 is selected from R or S, wherein X ⁶ is selected from S or T, wherein X ⁷ is selected from G or D, wherein X ⁸ is selected from Q or R, wherein X ⁹ is selected from Q, R, or K, wherein X ¹⁰ is selected from T or P, wherein X 11 is selected from N or Q, wherein X 12 is selected from S or A, wherein X ¹³ is selected from M or L, wherein X ¹⁴ is selected from N or Q, and wherein X ¹⁵ is selected from V or S.

[0072] In some embodiments, a peptide comprises the sequence

GSX ¹ X ² X ³ VX ⁴ IX ⁵ VX ⁶ CX ⁷ X ⁸ SX ⁹ X ¹⁰ CLX ¹¹ PCRX ¹² AGMRFGRCX ¹³ NX ¹⁴ RCX ¹⁵ CTPX ¹⁶ (SEQ ID NO: 15) or

χ ^! χ ² χ ³ VX ⁴ IX ⁵ VX ⁶ CX ⁷ X ⁸ SX ⁹ X ¹⁰ CLX ¹ ^CRX^AGMRFGRCX^NX^RCX^CTPX ¹⁶ (SEQ ID NO: 289), wherein X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ , X ⁹ , X ¹⁰ , X ¹¹ , X ¹² , X ¹³ , X ¹⁴ , X ¹⁵ , X ¹⁶ are each individually any amino acid or amino acid analogue or null. In some cases, the peptide comprises the sequence

GSX ¹ X ² X ³ VX ⁴ IX ⁵ VX ⁶ CX ⁷ X ⁸ SX ⁹ X ¹⁰ CLX ¹¹ PCRX ¹² AGMRFGRCX ¹³ NX ¹⁴ RCX ¹⁵ CTPX ¹⁶ (SEQ ID NO: 16) or

χ ^! χ ² χ ³ VX ⁴ IX ⁵ VX ⁶ CX ⁷ X ⁸ SX ⁹ X ¹⁰ CLX ¹ ^CRX^AGMRFGRCX^NX^RCX^CTPX ¹⁶ (SEQ ID

NO: 290), where X 1 is selected from G or null, wherein X 2 is selected from G, S or null, wherein

X 3 is selected from G, S or null, wherein X 4 is selected from P or R, wherein X 5 is selected from

N or P, wherein X 6 is selected from R, K or S, wherein X 7 is selected from R or K, wherein X 8 is selected from G or H, wherein X ⁹ is selected from R or G, wherein X ¹⁰ is selected from D or Q, wherein X 11 is selected from D, R, or K, wherein X 12 is selected from K, R, or D, wherein X 13 is selected from I or M, wherein X ¹⁴ is selected from S or G, wherein X ¹⁵ is selected from H or D, and wherein X ¹⁶ is selected from K, R, or null.

[0073] In some embodiments, a peptide comprises the sequence

GSXVXVRCXGSRQCXPCRRXGXRXGRCINRRXCRCYX (SEQ ID NO: 17) or

GSXGCVXRCRPGXRXCCXPXRRCSRRFGXRRCRX (SEQ ID NO: 18), wherein each letter is each individually any amino acid or amino acid analogue and where X is no amino acid or a 1-

10 amino acid long peptide fragment wherein each amino acid within such peptide fragment can in each case be any amino acid or amino acid analogue. In some embodiments, a peptide comprises the sequence XVXVRCXGSRQCXPCRRXGXRXGRCINRRXCRCYX (SEQ ID NO: 291) or XGCVXRCRPGXRXCCXPXRRCSRRFGXRRCRX (SEQ ID NO: 292), wherein each letter is each individually any amino acid or amino acid analogue and where X is no amino acid or a 1-10 amino acid long peptide fragment wherein each amino acid within such peptide fragment can in each case be any amino acid or amino acid analogue.

[0074] In some embodiments, a peptide comprises the sequence

GSXVXXXVRCXGSRQCXXPCRRXXGXRXGRCINRRXCRCYXXX (SEQ ID NO: 19), XVXXXVRCXGSRQCXXPCRRXXGXRXGRCINRRXCRCYXXX (SEQ ID NO: 293), GSXXXGCVXXXXRCRPGXRXCCXPXRRCSRRFGXXXXRRCRXXXXXX (SEQ ID NO: 20), or XXXGCVXXXXRCRPGXRXCCXPXRRCSRRFGXXXXRRCRXXXXXX (SEQ ID NO: 294) wherein X is no amino acid or any amino acid analogue.

[0075] In some embodiments, a peptide comprises the one or more of the following peptide fragments: GKCMNGKC (SEQ ID NO: 312); GRCMNGRC (SEQ ID NO: 313);

GKCINKKCKC (SEQ ID NO: 298); KCIN (SEQ ID NO: 299); KKCK (SEQ ID NO: 300); PCKR (SEQ ID NO: 301); KRCSRR (SEQ ID NO: 302); KQC (SEQ ID NO: 303);

GRCINRRCRC (SEQ ID NO: 304); RCIN (SEQ ID NO: 305); RRCR (SEQ ID NO: 306); PCRR (SEQ ID NO: 307); RRCSRR (SEQ ID NO: 308); RQC (SEQ ID NO: 309); PCKK (SEQ ID NO: 310), and KKCSKK (SEQ ID NO: 311).

[0076] TABLE 1 lists some exemplary peptides according to the present disclosure.

TABLE 1 - Exemplary Amino Acid Sequences

SEQ ID NO: Amino Acid Sequence

GSMNTKFIFLLLVVTNTMMLFDTKPVEGISCTGSKQCYDPCKRKTGCPNAK

SEQ ID NO: 40 CMNKSCKCYGCG

SEQ ID NO: 41 GSGVPINVKCSGSRDCLEPCKKAGMRFGKCINRKCHCTPK

SEQ ID NO: 42 GSGVPINVKCTGSPQCLKPCKDAGMRFGKCINGKCHCTPK

SEQ ID NO: 43 GSGVIINVKCKISRQCLEPCKKAGMRFGKCMNGKCHCTPK

SEQ ID NO: 44 GSGVPINVKCRGSPQCIQPCRDAGMRFGKCMNGKCHCTPQ

SEQ ID NO: 45 GSGVEINVKCTGSHQCIKPCKDAGMRFGKCINRKCHCTPK

SEQ ID NO: 46 GSGVEINVKCSGSPQCLKPCKDAGMRFGKCMNRKCHCTPK

SEQ ID NO: 47 GSGVPTDVKCRGSPQCIQPCKDAGMRFGKCMNGKCHCTPK

SEQ ID NO: 48 GSGVPINVSCTGSPQCIKPCKDAGMRFGKCMNRKCHCTPK

SEQ ID NO: 49 GSGVPINVPCTGSPQCIKPCKDAGMRFGKCMNRKCHCTPK

SEQ ID NO: 50 GSVGINVKCKHSGQCLKPCKDAGMRFGKCINGKCDCTPK

SEQ ID NO: 51 GSVGINVKCKHSGQCLKPCKDAGMRFGKCMNGKCDCTPK

SEQ ID NO: 52 GSVGIPVSCKHSGQCIKPCKDAGMRFGKCMNRKCDCTPK

SEQ ID NO: 53 GSRKGCFKEGHSCPKTAPCCRPLVCKGPSPNTKKCTRP

SEQ ID NO: 54 GSSFCIPFKPCKSDENCCKKFKCKTTGIVKLCRW

SEQ ID NO: 55 GSLKGCLPRNRFCNALSGPRCCSGLRCKELSIWASKCL

SEQ ID NO: 56 GSGNYCLRGRCLPGGRKCCNGRPCECFAKICSCKPK

SEQ ID NO: 57 GSTVKCGGCNRKCCPGGCRSGKCINGKCQCY

SEQ ID NO: 58 GSGCMKEYCAGQCRGKVSQDYCLKHCKCIPR

SEQ ID NO: 59 GSACLGFGEKCNPSNDKCCKSSSLVCSQKHKWCKYG

SEQ ID NO: 60 GSRGGCLPHNRFCNALSGPRCCSGLRCKELSIRDSRCLG

SEQ ID NO: 61 GSRGGCLPRNKFCNPSSGPRCCSGLTCKELNIWASKCL

SEQ ID NO: 62 GSQRSCAKPGDMCMGIKCCDGQCGCNRGTGRCFCK

SEQ ID NO: 63 GSARGCADAYKSCNHPRTCCDGYNGYKRACICSGSNCKCKKS

SEQ ID NO: 64 GSRGGCLPHNRFCNALSGPRCCSGLRCKELSIWDSRCLG

SEQ ID NO: 65 GSRGGCLPHNRFCNALSGPRCCSGLKCKELSIYDSRCLG

SEQ ID NO: 66 GSRGGCLPHNRFCNALSGPRCCSRLKCKELSIWDSRCLG

SEQ ID NO: 67 GSRGGCLPHNRFCNALTGPRCCSRLRCKELSIWDSICLG

SEQ ID NO: 68 GSSCADAYKSCDSLKCCNNRTCMCSMIGTNCTCRKK

SEQ ID NO: 69 GSERRCLPAGKTCVRGPMRVPCCGSCSQNKCT

SEQ ID NO: 70 GSLCSREGEFCYKLRKCCAGFYCKAFVLHCYRN

SEQ ID NO: 71 GSACGSCRKKCKGSGKCINGRCKCY

SEQ ID NO: 72 GSACGSCRKKCKGPGKCINGRCKCY

SEQ ID NO: 73 GSACQGYMRKCGRDKPPCCKKLECSKTWRWCVWN

SEQ ID NO: 74 GSGRYCQKWMWTCDSKRACCEGLRCKLWCRKI

SEQ ID NO: 75 GSNAKCRGSPECLPKCKEAIGKAAGKCMNGKCKCYP

SEQ ID NO: 76 GSNVKCRGSKECLPACKAAVGKAAGKCMNGKCKCYP

SEQ ID NO: 77 GSNVKCRGSPECLPKCKEAIGKSAGKCMNGKCKCYP

SEQ ID NO: 78 GSNAKCRGSPECLPKCKQAIGKAAGKCMNGKCKCYP

SEQ ID NO: 79 GSRGYCAEKGIKCHNIHCCSGLTCKCKGSSCVCRK

SEQ ID NO: 80 GSERGCKLTFWKCKNKKECCGWNACALGICMPR

SEQ ID NO: 81 GSKKKCIAKDYGRCKWGGTPCCRGRGCICSIMGTNCECKPR

SEQ ID NO: 82 GSGCKLTFWKCKNKKECCGWNACALGICMPR

SEQ ID NO: 83 GSACKGLFVTCTPGKDECCPNHVCSSKHKWCKYK

SEQ ID NO: 84 GSIACAPRGLLCFRDKECCKGLTCKGRFVNTWPTFCLV SEQ ID NO: Amino Acid Sequence

SEQ ID NO 85 GSACAGLYKKCGKGVNTCCENRPCKCDLAMGNCICKKK

SEQ ID NO 86 GSFTCAISCDIKVNGKPCKGSGEKKCSGGWSCKFNVCVKV

SEQ ID NO 87 GSGFCAQKGIKCHDIHCCTNLKCVREGSNRVCRKA

SEQ ID NO 88 GSCAKKRNWCGKNEDCCCPMKCIYAWYNQQGSCQSTITGLFKKC

SEQ ID NO 89 GSYCQKWMWTCDSARKCCEGLVCRLWCKKI

SEQ ID NO 90 GSRGGCLPHNKFCNALSGPRCCSGLKCKELTIWNTKCLE

SEQ ID NO 91 GSNVKCTGSKQCLPACKAAVGKAAGKCMNGKCKCYT

SEQ ID NO 92 GSQRSCAKPGEMCMRIKCCDGQCGCNRGTGRCFCK

SEQ ID NO 93 GSGCIPKHKRCTWSGPKCCNNISCHCNISGTLCKCRPG

SEQ ID NO 94 GSNYCVAKRCRPGGRQCCSGKPCACVGKVCKCPRD

SEQ ID NO 95 GSERGCSGAYKRCSSSQRCCEGRPCVCSAINSNCKCRKT

SEQ ID NO 96 GSRYCPRNPEACYNYCLRTGRPGGYCGGRSRITCFCFR

SEQ ID NO 97 GSQRSCAKPGEMCMGIKCCDGQCGCNRGTGRCFCK

SEQ ID NO 98 GSRRGCFKEGKWCPKSAPCCAPLKCKGPSIKQQKCVRE

SEQ ID NO 99 GSTVKCGGCNRKCCAGGCRSGKCINGKCQCYGR

SEQ ID NO 100 GSERRCEPSGKPCRPLMRIPCCGSCVRGKCA

SEQ ID NO 101 GSRGGCLPRNKFCNPSSGPRCCSGLTCKELNIWANKCL

SEQ ID NO 102 GSCAKKRNWCGKNEDCCCPMKCIYAWYNQQGSCQTTITGLFKKC

SEQ ID NO 103 GSVRIPVSCKHSGQCLKPCKDAGMRTGKCMNGKCDCTPK

SEQ ID NO 104 GSVKCTTSKDCWPPCKKVTGRA

SEQ ID NO 105 GSGIVCRVCRIICGMQGRRVNICRAPIRCRCRRG

SEQ ID NO 106 GSSERDCIRHLQRCRENRDCCSRRCSRRGTNPERRCR

SEQ ID NO 107 GSVRIPVSCRHSGQCLRPCRDAGMRFGRCMNGRCDCTPR

SEQ ID NO 108 GSGVPINVRCRGSRDCLDPCRRAGMRFGRCINSRCHCTP

SEQ ID NO 109 GSAVCVYRTCDRDCRRRGYRSGRCINNACRCYPYG

SEQ ID NO 110 GSISCTGSRQCYDPCRRRTGCPNARCMNRSCRCYGCG

SEQ ID NO 111 GSQVQTNVRCQGGSCASVCRREIGVAAGRCINGRCVCYRN

SEQ ID NO 112 GSEVIRCSGSRQCYGPCRQQTGCTNSRCMNRVCRCYGCG

SEQ ID NO 113 GSACRGVFDACTPGRNECCPNRVCSDRHRWCRWRL

SEQ ID NO 114 GSQIYTSRECNGSSECYSHCEGITGRRSGRCINRRCYCYR

SEQ ID NO 115 GSGCLEFWWRCNPNDDRCCRPRLRCSRLFRLCNFSFG

SEQ ID NO 116 GSDCVRFWGRCSQTSDCCPHLACRSRWPRNICVWDGSVG

SEQ ID NO 117 GSGCFGYRCDYYRGCCSGYVCSPTWRWCVRPGPGR

GSMNARFILLLVLTTMMLLPDTRGAEVIRCSGSRQCYGPCRQQTGCTNSRC

SEQ ID NO: 118 MNRVCRCYGCG

GSMNARLIYLLLVVTTMTLMFDTAQAVDIMCSGPRQCYGPCRRETGCPNA

SEQ ID NO: 119 RCMNRRCRCYGCV

GSMNARLIYLLLVVTTMMLTFDTTQAGDIRCSGTRQCWGPCRRQTTCTNSR

SEQ ID NO: 120 CMNGRCRCYGCVG

GSMNTRFIFLLLVVTNTMMLFDTRPVEGISCTGSRQCYDPCRRRTGCPNARC

SEQ ID NO 121 MNRSCRCYGCG

SEQ ID NO 122 GSGVPINVRCSGSRDCLEPCRRAGMRFGRCINRRCHCTPR

SEQ ID NO 123 GSGVPINVRCTGSPQCLRPCRDAGMRFGRCINGRCHCTPR

SEQ ID NO 124 GSGVIINVRCRISRQCLEPCRRAGMRFGRCMNGRCHCTPR

SEQ ID NO 125 GSGVPINVRCRGSPQCIQPCRDAGMRFGRCMNGRCHCTPQ

SEQ ID NO 126 GSGVEINVRCTGSHQCIRPCRDAGMRFGRCINRRCHCTPR

SEQ ID NO 127 GSGVEINVRCSGSPQCLRPCRDAGMRFGRCMNRRCHCTPR SEQ ID NO: Amino Acid Sequence

SEQ ID NO 128 GSGVPTDVRCRGSPQCIQPCRDAGMRFGRCMNGRCHCTPR

SEQ ID NO 129 GSGVPINVSCTGSPQCIRPCRDAGMRFGRCMNRRCHCTPR

SEQ ID NO 130 GSGVPINVPCTGSPQCIRPCRDAGMRFGRCMNRRCHCTPR

SEQ ID NO 131 GSVGINVRCRHSGQCLRPCRDAGMRFGRCINGRCDCTPR

SEQ ID NO 132 GSVGINVRCRHSGQCLRPCRDAGMRFGRCMNGRCDCTPR

SEQ ID NO 133 GSVGIPVSCRHSGQCIRPCRDAGMRFGRCMNRRCDCTPR

SEQ ID NO 134 GSRRGCFREGHSCPRTAPCCRPLVCRGPSPNTRRCTRP

SEQ ID NO 135 GSSFCIPFRPCRSDENCCRRFRCRTTGIVRLCRW

SEQ ID NO 136 GSLRGCLPRNRFCNALSGPRCCSGLRCRELSIWASRCL

SEQ ID NO 137 GSGNYCLRGRCLPGGRRCCNGRPCECFARICSCRPR

SEQ ID NO 138 GSTVRCGGCNRRCCPGGCRSGRCINGRCQCY

SEQ ID NO 139 GSGCMREYCAGQCRGRVSQDYCLRHCRCIPR

SEQ ID NO 140 GSACLGFGERCNPSNDRCCRSSSLVCSQRHRWCRYG

SEQ ID NO 141 GSRGGCLPHNRFCNALSGPRCCSGLRCRELSIRDSRCLG

SEQ ID NO 142 GSRGGCLPRNRFCNPSSGPRCCSGLTCRELNIWASRCL

SEQ ID NO 143 GSQRSCARPGDMCMGIRCCDGQCGCNRGTGRCFCR

SEQ ID NO 144 GSARGCADAYRSCNHPRTCCDGYNGYRRACICSGSNCRCRRS

SEQ ID NO 145 GSRGGCLPHNRFCNALSGPRCCSGLRCRELSIWDSRCLG

SEQ ID NO 146 GSRGGCLPHNRFCNALSGPRCCSGLRCRELSIYDSRCLG

SEQ ID NO 147 GSRGGCLPHNRFCNALSGPRCCSRLRCRELSIWDSRCLG

SEQ ID NO 148 GSRGGCLPHNRFCNALTGPRCCSRLRCRELSIWDSICLG

SEQ ID NO 149 GSSCADAYKSCDSLRCCNNRTCMCSMIGTNCTCRRR

SEQ ID NO 150 GSERRCLPAGRTCVRGPMRVPCCGSCSQNRCT

SEQ ID NO 151 GSLCSREGEFCYRLRRCCAGFYCRAFVLHCYRN

SEQ ID NO 152 GSACGSCRRRCRGSGRCINGRCRCY

SEQ ID NO 153 GSACGSCRRRCRGPGRCINGRCRCY

SEQ ID NO 154 GSACQGYMRRCGRDRPPCCRRLECSRTWRWCVWN

SEQ ID NO 155 GSGRYCQRWMWTCDSRRACCEGLRCRLWCRRI

SEQ ID NO 156 GSNARCRGSPECLPRCREAIGRAAGRCMNGRCRCYP

SEQ ID NO 157 GSNVRCRGSRECLPACRAAVGRAAGRCMNGRCRCYP

SEQ ID NO 158 GSNVRCRGSPECLPRCREAIGRSAGRCMNGRCRCYP

SEQ ID NO 159 GSNARCRGSPECLPRCRQAIGRAAGRCMNGRCRCYP

SEQ ID NO 160 GSRGYCAERGIRCHNIHCCSGLTCRCRGSSCVCRR

SEQ ID NO 161 GSERGCRLTFWRCRNRRECCGWNACALGICMPR

SEQ ID NO 162 GSRRRCIARDYGRCRWGGTPCCRGRGCICSIMGTNCECRPR

SEQ ID NO 163 GSGCRLTFWRCRNRRECCGWNACALGICMPR

SEQ ID NO 164 GSACRGLFVTCTPGRDECCPNHVCSSRHRWCRYR

SEQ ID NO 165 GSIACAPRGLLCFRDRECCRGLTCRGRFVNTWPTFCLV

SEQ ID NO 166 GSACAGLYRRCGRGVNTCCENRPCRCDLAMGNCICRRR

SEQ ID NO 167 GSFTCAISCDIRVNGRPCRGSGERRCSGGWSCRFNVCVRV

SEQ ID NO 168 GSGFCAQRGIRCHDIHCCTNLRCVREGSNRVCRRA

SEQ ID NO 169 GSCARRRNWCGRNEDCCCPMRCIYAWYNQQGSCQSTITGLFRRC

SEQ ID NO 170 GSYCQRWMWTCDSARRCCEGLVCRLWCRRI

SEQ ID NO 171 GSRGGCLPHNRFCNALSGPRCCSGLRCRELTIWNTRCLE

SEQ ID NO 172 GSNVRCTGSRQCLPACRAAVGRAAGRCMNGRCRCYT SEQ ID NO: Amino Acid Sequence

SEQ ID NO 173 GSQRSCARPGEMCMRIRCCDGQCGCNRGTGRCFCR

SEQ ID NO 174 GSGCIPRHRRCTWSGPRCCNNISCHCNISGTLCRCRPG

SEQ ID NO 175 GSNYCVARRCRPGGRQCCSGRPCACVGRVCRCPRD

SEQ ID NO 176 GSERGCSGAYRRCSSSQRCCEGRPCVCSAINSNCRCRRT

SEQ ID NO 177 GSQRSCARPGEMCMGIRCCDGQCGCNRGTGRCFCR

SEQ ID NO 178 GSRRGCFREGRWCPRSAPCCAPLRCRGPSIRQQRCVRE

SEQ ID NO 179 GSTVRCGGCNRRCCAGGCRSGRCINGRCQCYGR

SEQ ID NO 180 GSERRCEPSGRPCRPLMRIPCCGSCVRGRCA

SEQ ID NO 181 GSRGGCLPRNRFCNPSSGPRCCSGLTCRELNIWANRCL

SEQ ID NO 182 GSCARRRNWCGRNEDCCCPMRCIYAWYNQQGSCQTTITGLFRRC

SEQ ID NO 183 GSVRIPVSCRHSGQCLRPCRDAGMRTGRCMNGRCDCTPR

SEQ ID NO 184 GSQKILSNRCNNSSECIPHCIRIFGTRAAKCINRKCYCYP

SEQ ID NO 185 GSAVCNLKRCQLSCRSLGLLGKCIGDKCECVKHG

SEQ ID NO 186 GSISIGIRCSPSIDLCEGQCRIRRYFTGYCSGDTCHCSG

SEQ ID NO 187 GSGDCLPHLRRCRENNDCCSRRCRRRGANPERRCR

SEQ ID NO 188 GSSCEPGRTFRDRCNTCKCGADGRSAACTLRACPNQ

SEQ ID NO 189 GSGDCLPHLKRCKADNDCCGKKCKRRGTNAEKRCR

SEQ ID NO 190 GSGDCLPHLKRCKENNDCCSKKCKRRGTNPEKRCR

SEQ ID NO 191 GSKDCLKKLKLCKENKDCCSKSCKRRGTNIEKRCR

SEQ ID NO 192 GSGDCLPHLKRCKENNDCCSKKCKRRGANPEKRCR

SEQ ID NO 193 GSVFINVKCRGSPECLPKCKEAIGKSAGKCMNGKCKCYP

SEQ ID NO 194 GSVFINAKCRGSPECLPKCKEAIGKAAGKCMNGKCKCYP

SEQ ID NO 195 GSVIINVKCKISRQCLEPCKKAGMRFGKCMNGKCHCTP

SEQ ID NO 196 GSVPTDVKCRGSPQCIQPCKDAGMRFGKCMNGKCHCTP

SEQ ID NO 197 GSVRIPVSCKHSGQCLKPCKDAGMRFGKCMNGKCDCTP

SEQ ID NO 198 GSVRIPVSCRHSGQCLRPCRDAGMRFGRCMNGRCDCTP

SEQ ID NO 199 GSTNVSCTTSKECWSVCQRLHNTSRGKCMNKKCRC

SEQ ID NO 200 GSNVKCTGSKQCLPACKAAVGKAAGKCMNGKCKC

SEQ ID NO 201 GSGVPINVRCRGSRDCLDPCRGAGERHGRCGNSRCHCTP

SEQ ID NO 202 GSVRIPVSCRHSGQCLRPCRDAGERHGRCGGGRCDCTPR

SEQ ID NO 203 GSQVQTNVRCQGGSCGSVCRREGGGAGGGCGNGRCGCYRN

SEQ ID NO 204 GSIKCSESYQCFPVCKSRFGKTNGRCVNGFCDCF

SEQ ID NO 205 GSVKCSSPQQCLKPCKAAFGISAGGKCINGKCKCY

SEQ ID NO 206 GSVSCSASSQCWPVCKKLFGTYRGKCMNSKCRCY

SEQ ID NO 207 GSESCTASNQCWSICKRLHNTNRGKCMNKKCRCY

SEQ ID NO 208 GSVSCTTSKECWSVCEKLYNTSRGKCMNKKCRCY

SEQ ID NO 209 GSMRCKSSKECLVKCKQATGRPNGKCMNRKCKCY

SEQ ID NO 210 GSIKCTLSKDCYSPCKKETGCPRAKCINRNCKCY

SEQ ID NO 211 GSIRCSGSRDCYSPCMKQTGCPNAKCINKSCKCY

SEQ ID NO 212 GSIRCSGTRECYAPCQKLTGCLNAKCMNKACKCY

SEQ ID NO 213 GSISCTNPKQCYPHCKKETGYPNAKCMNRKCKCF

SEQ ID NO 214 GSASCRTPKDCADPCRKETGCPYGKCMNRKCKCN

SEQ ID NO 215 GSTSCISPKQCTEPCRAKGCKHGKCMNRKCHCM

SEQ ID NO 216 GSKECTGPQHCTNFCRKN-KCTHGKCMNRKCKCF

SEQ ID NO 217 GSIKCRTPKDCADPCRKQTGCPHAKCMNKTCRCH SEQ ID NO: Amino Acid Sequence

SEQ ID NO: 218 GSVKCTTSKECWPPCKAATGKAAGKCMNKKCKCQ

SEQ ID NO: 219 GSLECGASRECYDPCFKAFGRAHGKCMNNKCRCY

SEQ ID NO: 220 GSEKCFATSQCWTPCKKAIGSLQSKCMNGKCKCY

SEQ ID NO: 221 GSVRCYASRECWEPCRRVTGSAQAKCQNNQCRCY

SEQ ID NO: 222 GSVKCSASRECWVACKKVTGSGQGKCQNNQCRCY

SEQ ID NO: 223 GSVKCISSQECWIACKKVTGRFEGKCQNRQCRCY

SEQ ID NO: 224 GSVRCYDSRQCWIACKKVTGSTQGKCQNKQCRCY

SEQ ID NO: 225 GSVDCTVSKECWAPCKAAFGVDRGKCMGKKCKCY

SEQ ID NO: 226 GSAKCRGSPECLPKCKEAIGKAAGKCMNGKCKCY

SEQ ID NO: 227 GSKKCQGGSCASVCRRVIGVAAGKCINGRCVCY

SEQ ID NO: 228 GSKKCSNTSQCYKTCEKVVGVAAGKCMNGKCICY

SEQ ID NO: 229 GSVKCSGSSKCVKICIDRYNTRGAKCINGRCTCY

SEQ ID NO: 230 GSNRCNNSSECIPHCIRIFGTRAAKCINRKCYCY

SEQ ID NO: 231 GSKECNGSSECYSHCEGITGKRSGKCINKKCYCY

SEQ ID NO: 232 GSAFCNLRRCELSCRSLGLLGKCIGEECKCV

SEQ ID NO: 233 GSAVCNLKRCQLSCRSLGLLGKCIGDKCECV

SEQ ID NO: 234 GSAACYSS-DCRVKCVAMGFSSGKCINSKCKCY

SEQ ID NO: 235 GSAICATDADCSRKCPGNPPCRNGFCACT

SEQ ID NO: 236 GSTECQIKNDCQRYCQSVKECKYGKCYCN

SEQ ID NO: 237 GSTQCQSVRDCQQYCLTPDRCSYGTCYCK

SEQ ID NO: 238 GSVSCRYGSDCAEPCKRLKCLLPSKCINGKCTCY

SEQ ID NO: 239 GSIKCRYPADCHIMCRKVTGRAEGKCMNGKCTCY

SEQ ID NO: 240 GSIKCSSSSSCYEPCRGVTGRAHGKCMNGRCTCY

SEQ ID NO: 241 GSVKCTGSKQCLPACKAAVGKAAGKCMNGKCKCY

SEQ ID NO: 242 GSVSCKHSGQCIKPCKDA-GMRFGKCMNRKCDCT

SEQ ID NO: 243 GSVKCRGSPQCIQPCRDA-GMRFGKCMNGKCHCT

SEQ ID NO: 244 GSVKCTSPKQCLPPCKAQFGIRAGAKCMNGKCKCY

SEQ ID NO: 245 GSVKCTSPKQCSKPCKELYGSSAGAKCMNGKCKCY

SEQ ID NO: 246 GSVKCTSPKQCLPPCKEIYGRHAGAKCMNGKCHCS

SEQ ID NO: 247 GSVKCTGSKQCWPVCKQMFGKPNGKCMNGKCRCY

SEQ ID NO: 248 GSVKCRGSRDCLDPCKKAGMRFGKCINSKCHCT

SEQ ID NO: 249 GSVRCVTDDDCFRKCPGNPSCKRGFCACK

SEQ ID NO: 250 GSVPCNNSRPCVPVCIREVNNKNGKCSNGKCLCY

SEQ ID NO: 251 GSVPINVKCRGSRDCLDPCKKAGMRFGKCINSKCHCTP

SEQ ID NO: 252 GSVQTNVKCQGGSCASVCRREIGVAAGKCINGKCVCYRN

SEQ ID NO: 253 GSAEIIRCSGTRECYAPCQKLTGCLNAKCMNKACKCYGCV

SEQ ID NO: 254 GSRPTDIKCSASYQCFPVCKSRFGKTNGRCVNGLCDCF

SEQ ID NO: 255 GSQFTDVKCTGSKQCWPVCKQMFGKPNGKCMNGKCRCYS

SEQ ID NO: 256 GSVGINVKCKHSRQCLKPCKDAGMRFGKCTNGKCHCTPK

SEQ ID NO: 257 GSVVIGQRCYRSPDCYSACKKLVGKATGKCTNGRCDC

SEQ ID NO: 258 GSNFKVEGACSKPCRKYCIDKGARNGKCINGRCHCYY

SEQ ID NO: 259 GSQIDTNVKCSGSSKCVKICIDRYNTRGAKCINGRCTCYP

SEQ ID NO: 260 GSGVPISVRCRGSRDCLEPCRRAGTRFGRCINGRCHCTP

SEQ ID NO: 261 GSGVPISVRCRGSRDCLEPCRRAGTRFGRCIQSRCHCTP

SEQ ID NO: 262 GSGVPISVRCRGSRDCLEPCRRAGTRFGRCINRRCHCTP SEQ ID NO: Amino Acid Sequence

SEQ ID NO: 263 GSGVPINVRCRGSRDCLEPCRRAGTRFGRCINSRCHCTP

SEQ ID NO: 264 GSGVPINVRCRGSRDCLEPCRRAGTRFGRCIQSRCHCTP

SEQ ID NO: 265 GSGVPINVRCRGSRDCLEPCRRAGTRFGRCIQSRCHCYP

SEQ ID NO: 266 GSGVPINVRCRGSRDCYEPCRRAGTRFGRCIQSRCHCTP

SEQ ID NO: 267 GSGVPINVRCRGSRDCLEPCRRAGTRFGRCIQSRCYCTP

SEQ ID NO: 268 GSGVPISVRCRGSRDCLEPCRRAGTRFGRCIQSRCHCYP

SEQ ID NO: 269 GSGVPISVRCRGSRDCYEPCRRAGTRFGRCIQSRCHCTP

SEQ ID NO: 270 GSGVPISVRCRGSRDCLEPCRRAGTRFGRCIQSRCYCTP

SEQ ID NO: 271 GSGVPINVRCRGSRDCLEPCRRAGTRFGRCIASRCHCYP

SEQ ID NO: 272 GSGVPINVRCRGSRDCLEPCRRAGTRFGRCISSRCHCYP

SEQ ID NO: 273 GSGVPINVRCRGSRDCLEPCRRAGTRFGRCITSRCHCYP

SEQ ID NO: 274 GSGVPINVRCRGSRDCLEPCRRAGTRFGRCINSRCHCYP

SEQ ID NO: 314 GIVCKVCKIICGMQGKKVNICKAPIKCKCKKG

SEQ ID NO: 315 SEKDCIKHLQRCRENKDCCSKKCSRRGTNPEKRCR

SEQ ID NO: 316 VRIPVSCKHSGQCLKPCKDAGMRFGKCMNGKCDCTPK

SEQ ID NO: 317 GVPINVKCRGSRDCLDPCKKAGMRFGKCINSKCHCTP

SEQ ID NO: 318 AVCVYRTCDKDCKRRGYRSGKCINNACKCYPYG

SEQ ID NO: 319 ISCTGSKQCYDPCKRKTGCPNAKCMNKSCKCYGCG

SEQ ID NO: 320 QVQTNVKCQGGSCASVCRREIGVAAGKCINGKCVCYRN

SEQ ID NO: 321 EVIRCSGSKQCYGPCKQQTGCTNSKCMNKVCKCYGCG

SEQ ID NO: 322 ACKGVFDACTPGKNECCPNRVCSDKHKWCKWKL

SEQ ID NO: 323 QIYTSKECNGSSECYSHCEGITGKRSGKCINKKCYCYR

SEQ ID NO: 324 GCLEFWWKCNPNDDKCCRPKLKCSKLFKLCNFSFG

SEQ ID NO: 325 DCVRFWGKCSQTSDCCPHLACKSKWPRNICVWDGSVG

SEQ ID NO: 326 GCFGYKCDYYKGCCSGYVCSPTWKWCVRPGPGR

MNAKFILLLVLTTMMLLPDTKGAEVIRCSGSKQCYGPCKQQTGCTNSKCM

SEQ ID NO: 327 NKVCKCYGCG

MNAKLIYLLLVVTTMTLMFDTAQAVDIMCSGPKQCYGPCKKETGCPNAKC

SEQ ID NO: 328 MNRRCKCYGCV

MNAKLIYLLLVVTTMMLTFDTTQAGDIKCSGTRQCWGPCKKQTTCTNSKC

SEQ ID NO: 329 MNGKCKCYGCVG

MNTKFIFLLLVVTNTMMLFDTKPVEGISCTGSKQCYDPCKRKTGCPNAKCM

SEQ ID NO: 330 NKSCKCYGCG

SEQ ID NO: 331 GVPINVKCSGSRDCLEPCKKAGMRFGKCINRKCHCTPK

SEQ ID NO: 332 GVPINVKCTGSPQCLKPCKDAGMRFGKCINGKCHCTPK

SEQ ID NO: 333 GVIINVKCKISRQCLEPCKKAGMRFGKCMNGKCHCTPK

SEQ ID NO: 334 GVPINVKCRGSPQCIQPCRDAGMRFGKCMNGKCHCTPQ

SEQ ID NO: 335 GVEINVKCTGSHQCIKPCKDAGMRFGKCINRKCHCTPK

SEQ ID NO: 336 GVEINVKCSGSPQCLKPCKDAGMRFGKCMNRKCHCTPK

SEQ ID NO: 337 GVPTDVKCRGSPQCIQPCKDAGMRFGKCMNGKCHCTPK

GVPINVSCTGSPQCIKPCKDAGMRFGKCMNRKCHCTPK

SEQ ID NO: 338

SEQ ID NO: 339 GVPINVPCTGSPQCIKPCKDAGMRFGKCMNRKCHCTPK

SEQ ID NO: 340 VGINVKCKHSGQCLKPCKDAGMRFGKCINGKCDCTPK

SEQ ID NO: 341 VGINVKCKHSGQCLKPCKDAGMRFGKCMNGKCDCTPK SEQ ID NO: Amino Acid Sequence

SEQ ID NO: 342 VGIPVSCKHSGQCIKPCKDAGMRFGKCMNRKCDCTPK

SEQ ID NO: 343 RKGCFKEGHSCPKTAPCCRPLVCKGPSPNTKKCTRP

SEQ ID NO: 344 SFCIPFKPCKSDENCCKKFKCKTTGIVKLCRW

SEQ ID NO: 345 LKGCLPRNRFCNALSGPRCCSGLRCKELSIWASKCL

SEQ ID NO: 346 GNYCLRGRCLPGGRKCCNGRPCECFAKICSCKPK

SEQ ID NO: 347 TVKCGGCNRKCCPGGCRSGKCINGKCQCY

SEQ ID NO: 348 GCMKEYCAGQCRGKVSQDYCLKHCKCIPR

SEQ ID NO: 349 ACLGFGEKCNPSNDKCCKSSSLVCSQKHKWCKYG

SEQ ID NO: 350 RGGCLPHNRFCNALSGPRCCSGLRCKELSIRDSRCLG

SEQ ID NO: 351 RGGCLPRNKFCNPSSGPRCCSGLTCKELNIWASKCL

SEQ ID NO: 352 QRSCAKPGDMCMGIKCCDGQCGCNRGTGRCFCK

SEQ ID NO: 353 ARGCADAYKSCNHPRTCCDGYNGYKRACICSGSNCKCKKS

SEQ ID NO: 354 RGGCLPHNRFCNALSGPRCCSGLRCKELSIWDSRCLG

SEQ ID NO: 355 RGGCLPHNRFCNALSGPRCCSGLKCKELSIYDSRCLG

SEQ ID NO: 356 RGGCLPHNRFCNALSGPRCCSRLKCKELSIWDSRCLG

SEQ ID NO: 357 RGGCLPHNRFCNALTGPRCCSRLRCKELSIWDSICLG

SEQ ID NO: 358 SCADAYKSCDSLKCCNNRTCMCSMIGTNCTCRKK

SEQ ID NO: 359 ERRCLPAGKTCVRGPMRVPCCGSCSQNKCT

SEQ ID NO: 360 LCSREGEFCYKLRKCCAGFYCKAFVLHCYRN

SEQ ID NO: 361 ACGSCRKKCKGSGKCINGRCKCY

SEQ ID NO: 362 ACGSCRKKCKGPGKCINGRCKCY

SEQ ID NO: 363 ACQGYMRKCGRDKPPCCKKLECSKTWRWCVWN

SEQ ID NO: 364 GRYCQKWMWTCDSKRACCEGLRCKLWCRKI

SEQ ID NO: 365 NAKCRGSPECLPKCKEAIGKAAGKCMNGKCKCYP

SEQ ID NO: 366 NVKCRGSKECLPACKAAVGKAAGKCMNGKCKCYP

SEQ ID NO: 367 NVKCRGSPECLPKCKEAIGKSAGKCMNGKCKCYP

SEQ ID NO: 368 NAKCRGSPECLPKCKQAIGKAAGKCMNGKCKCYP

SEQ ID NO: 369 RGYCAEKGIKCHNIHCCSGLTCKCKGSSCVCRK

SEQ ID NO: 370 ERGCKLTFWKCKNKKECCGWNACALGICMPR

SEQ ID NO: 371 KKKCIAKDYGRCKWGGTPCCRGRGCICSIMGTNCECKPR

SEQ ID NO: 372 GCKLTFWKCKNKKECCGWNACALGICMPR

SEQ ID NO: 373 ACKGLFVTCTPGKDECCPNHVCSSKHKWCKYK

SEQ ID NO: 374 IACAPRGLLCFRDKECCKGLTCKGRFVNTWPTFCLV

SEQ ID NO: 375 ACAGLYKKCGKGVNTCCENRPCKCDLAMGNCICKKK

SEQ ID NO: 376 FTCAISCDIKVNGKPCKGSGEKKCSGGWSCKFNVCVKV

SEQ ID NO: 377 GFCAQKGIKCHDIHCCTNLKCVREGSNRVCRKA

SEQ ID NO: 378 CAKKRNWCGKNEDCCCPMKCIYAWYNQQGSCQSTITGLFKKC

SEQ ID NO: 379 YCQKWMWTCDSARKCCEGLVCRLWCKKI

SEQ ID NO: 380 RGGCLPHNKFCNALSGPRCCSGLKCKELTIWNTKCLE

SEQ ID NO: 381 NVKCTGSKQCLPACKAAVGKAAGKCMNGKCKCYT

SEQ ID NO: 382 QRSCAKPGEMCMRIKCCDGQCGCNRGTGRCFCK

SEQ ID NO: 383 GCIPKHKRCTWSGPKCCNNISCHCNISGTLCKCRPG

SEQ ID NO: 384 NYCVAKRCRPGGRQCCSGKPCACVGKVCKCPRD

SEQ ID NO: 385 ERGCSGAYKRCSSSQRCCEGRPCVCSAINSNCKCRKT SEQ ID NO: Amino Acid Sequence

SEQ ID NO: 386 RYCPRNPEACYNYCLRTGRPGGYCGGRSRITCFCFR

SEQ ID NO: 387 QRSCAKPGEMCMGIKCCDGQCGCNRGTGRCFCK

SEQ ID NO: 388 RRGCFKEGKWCPKSAPCCAPLKCKGPSIKQQKCVRE

SEQ ID NO: 389 TVKCGGCNRKCCAGGCRSGKCINGKCQCYGR

SEQ ID NO: 390 ERRCEPSGKPCRPLMRIPCCGSCVRGKCA

SEQ ID NO: 391 RGGCLPRNKFCNPSSGPRCCSGLTCKELNIWANKCL

SEQ ID NO: 392 CAKKRNWCGKNEDCCCPMKCIYAWYNQQGSCQTTITGLFKKC

SEQ ID NO: 393 VRIPVSCKHSGQCLKPCKDAGMRTGKCMNGKCDCTPK

SEQ ID NO: 394 VKCTTSKDCWPPCKKVTGRA

SEQ ID NO: 395 GIVCRVCRIICGMQGRRVNICRAPIRCRCRRG

SEQ ID NO: 396 SERDCIRHLQRCRENRDCCSRRCSRRGTNPERRCR

SEQ ID NO: 397 VRIPVSCRHSGQCLRPCRDAGMRFGRCMNGRCDCTPR

SEQ ID NO: 398 GVPINVRCRGSRDCLDPCRRAGMRFGRCINSRCHCTP

SEQ ID NO: 399 AVCVYRTCDRDCRRRGYRSGRCINNACRCYPYG

SEQ ID NO: 400 ISCTGSRQCYDPCRRRTGCPNARCMNRSCRCYGCG

SEQ ID NO: 401 QVQTNVRCQGGSCASVCRREIGVAAGRCINGRCVCYRN

SEQ ID NO: 402 EVIRCSGSRQCYGPCRQQTGCTNSRCMNRVCRCYGCG

SEQ ID NO: 403 ACRGVFDACTPGRNECCPNRVCSDRHRWCRWRL

SEQ ID NO: 404 QIYTSRECNGSSECYSHCEGITGRRSGRCINRRCYCYR

SEQ ID NO: 405 GCLEFWWRCNPNDDRCCRPRLRCSRLFRLCNFSFG

SEQ ID NO: 406 DCVRFWGRCSQTSDCCPHLACRSRWPRNICVWDGSVG

SEQ ID NO: 407 GCFGYRCDYYRGCCSGYVCSPTWRWCVRPGPGR

MNARFILLLVLTTMMLLPDTRGAEVIRCSGSRQCYGPCRQQTGCTNSRCMN

SEQ ID NO: 408 RVCRCYGCG

MNARLIYLLLVVTTMTLMFDTAQAVDIMCSGPRQCYGPCRRETGCPNARC

SEQ ID NO: 409 MNRRCRCYGCV

MNARLIYLLLVVTTMMLTFDTTQAGDIRCSGTRQCWGPCRRQTTCTNSRC

SEQ ID NO: 410 MNGRCRCYGCVG

MNTRFIFLLLVVTNTMMLFDTRPVEGISCTGSRQCYDPCRRRTGCPNARCM

SEQ ID NO: 411 NRSCRCYGCG

SEQ ID NO: 412 GVPINVRCSGSRDCLEPCRRAGMRFGRCINRRCHCTPR

SEQ ID NO: 413 GVPINVRCTGSPQCLRPCRDAGMRFGRCINGRCHCTPR

SEQ ID NO: 414 GVIINVRCRISRQCLEPCRRAGMRFGRCMNGRCHCTPR

SEQ ID NO: 415 GVPINVRCRGSPQCIQPCRDAGMRFGRCMNGRCHCTPQ

SEQ ID NO: 416 GVEINVRCTGSHQCIRPCRDAGMRFGRCINRRCHCTPR

SEQ ID NO: 417 GVEINVRCSGSPQCLRPCRDAGMRFGRCMNRRCHCTPR

SEQ ID NO: 418 GVPTDVRCRGSPQCIQPCRDAGMRFGRCMNGRCHCTPR

SEQ ID NO: 419 GVPINVSCTGSPQCIRPCRDAGMRFGRCMNRRCHCTPR

SEQ ID NO: 420 GVPINVPCTGSPQCIRPCRDAGMRFGRCMNRRCHCTPR

SEQ ID NO: 421 VGINVRCRHSGQCLRPCRDAGMRFGRCINGRCDCTPR

SEQ ID NO: 422 VGINVRCRHSGQCLRPCRDAGMRFGRCMNGRCDCTPR

SEQ ID NO: 423 VGIPVSCRHSGQCIRPCRDAGMRFGRCMNRRCDCTPR

SEQ ID NO: 424 RRGCFREGHSCPRTAPCCRPLVCRGPSPNTRRCTRP

SEQ ID NO: 425 SFCIPFRPCRSDENCCRRFRCRTTGIVRLCRW

SEQ ID NO: 426 LRGCLPRNRFCNALSGPRCCSGLRCRELSIWASRCL SEQ ID NO: Amino Acid Sequence

SEQ ID NO: 427 GNYCLRGRCLPGGRRCCNGRPCECFARICSCRPR

SEQ ID NO: 428 TVRCGGCNRRCCPGGCRSGRCINGRCQCY

SEQ ID NO: 429 GCMREYCAGQCRGRVSQDYCLRHCRCIPR

SEQ ID NO: 430 ACLGFGERCNPSNDRCCRSSSLVCSQRHRWCRYG

SEQ ID NO: 431 RGGCLPHNRFCNALSGPRCCSGLRCRELSIRDSRCLG

SEQ ID NO: 432 RGGCLPRNRFCNPSSGPRCCSGLTCRELNIWASRCL

SEQ ID NO: 433 QRSCARPGDMCMGIRCCDGQCGCNRGTGRCFCR

SEQ ID NO: 434 ARGCADAYRSCNHPRTCCDGYNGYRRACICSGSNCRCRRS

SEQ ID NO: 435 RGGCLPHNRFCNALSGPRCCSGLRCRELSIWDSRCLG

SEQ ID NO: 436 RGGCLPHNRFCNALSGPRCCSGLRCRELSIYDSRCLG

SEQ ID NO: 437 RGGCLPHNRFCNALSGPRCCSRLRCRELSIWDSRCLG

SEQ ID NO: 438 RGGCLPHNRFCNALTGPRCCSRLRCRELSIWDSICLG

SEQ ID NO: 439 SCADAYKSCDSLRCCNNRTCMCSMIGTNCTCRRR

SEQ ID NO: 440 ERRCLPAGRTCVRGPMRVPCCGSCSQNRCT

SEQ ID NO: 441 LCSREGEFCYRLRRCCAGFYCRAFVLHCYRN

SEQ ID NO: 442 ACGSCRRRCRGSGRCINGRCRCY

SEQ ID NO: 443 ACGSCRRRCRGPGRCINGRCRCY

SEQ ID NO: 444 ACQGYMRRCGRDRPPCCRRLECSRTWRWCVWN

SEQ ID NO: 445 GRYCQRWMWTCDSRRACCEGLRCRLWCRRI

SEQ ID NO: 446 NARCRGSPECLPRCREAIGRAAGRCMNGRCRCYP

SEQ ID NO: 447 NVRCRGSRECLPACRAAVGRAAGRCMNGRCRCYP

SEQ ID NO: 448 NVRCRGSPECLPRCREAIGRSAGRCMNGRCRCYP

SEQ ID NO: 449 NARCRGSPECLPRCRQAIGRAAGRCMNGRCRCYP

SEQ ID NO: 450 RGYCAERGIRCHNIHCCSGLTCRCRGSSCVCRR

SEQ ID NO: 451 ERGCRLTFWRCRNRRECCGWNACALGICMPR

SEQ ID NO: 452 RRRCIARDYGRCRWGGTPCCRGRGCICSIMGTNCECRPR

SEQ ID NO: 453 GCRLTFWRCRNRRECCGWNACALGICMPR

SEQ ID NO: 454 ACRGLFVTCTPGRDECCPNHVCSSRHRWCRYR

SEQ ID NO: 455 IACAPRGLLCFRDRECCRGLTCRGRFVNTWPTFCLV

SEQ ID NO: 456 ACAGLYRRCGRGVNTCCENRPCRCDLAMGNCICRRR

SEQ ID NO: 457 FTCAISCDIRVNGRPCRGSGERRCSGGWSCRFNVCVRV

SEQ ID NO: 458 GFCAQRGIRCHDIHCCTNLRCVREGSNRVCRRA

SEQ ID NO: 459 CARRRNWCGRNEDCCCPMRCIYAWYNQQGSCQSTITGLFRRC

SEQ ID NO: 460 YCQRWMWTCDSARRCCEGLVCRLWCRRI

SEQ ID NO: 461 RGGCLPHNRFCNALSGPRCCSGLRCRELTIWNTRCLE

SEQ ID NO: 462 NVRCTGSRQCLPACRAAVGRAAGRCMNGRCRCYT

SEQ ID NO: 463 QRSCARPGEMCMRIRCCDGQCGCNRGTGRCFCR

SEQ ID NO: 464 GCIPRHRRCTWSGPRCCNNISCHCNISGTLCRCRPG

SEQ ID NO: 465 NYCVARRCRPGGRQCCSGRPCACVGRVCRCPRD

SEQ ID NO: 466 ERGCSGAYRRCSSSQRCCEGRPCVCSAINSNCRCRRT

SEQ ID NO: 467 QRSCARPGEMCMGIRCCDGQCGCNRGTGRCFCR

SEQ ID NO: 468 RRGCFREGRWCPRSAPCCAPLRCRGPSIRQQRCVRE

SEQ ID NO: 469 TVRCGGCNRRCCAGGCRSGRCINGRCQCYGR

SEQ ID NO: 470 ERRCEPSGRPCRPLMRIPCCGSCVRGRCA

SEQ ID NO: 471 RGGCLPRNRFCNPSSGPRCCSGLTCRELNIWANRCL SEQ ID NO: Amino Acid Sequence

SEQ ID NO: 472 CARRRNWCGRNEDCCCPMRCIYAWYNQQGSCQTTITGLFRRC

SEQ ID NO: 473 VRIPVSCRHSGQCLRPCRDAGMRTGRCMNGRCDCTPR

SEQ ID NO: 474 QKILSNRCNNSSECIPHCIRIFGTRAAKCINRKCYCYP

SEQ ID NO: 475 AVCNLKRCQLSCRSLGLLGKCIGDKCECVKHG

SEQ ID NO: 476 ISIGIRCSPSIDLCEGQCRIRRYFTGYCSGDTCHCSG

SEQ ID NO: 477 GDCLPHLRRCRENNDCCSRRCRRRGANPERRCR

SEQ ID NO: 478 SCEPGRTFRDRCNTCKCGADGRSAACTLRACPNQ

SEQ ID NO: 479 GDCLPHLKRCKADNDCCGKKCKRRGTNAEKRCR

SEQ ID NO: 480 GDCLPHLKRCKENNDCCSKKCKRRGTNPEKRCR

SEQ ID NO: 481 KDCLKKLKLCKENKDCCSKSCKRRGTNIEKRCR

SEQ ID NO: 482 GDCLPHLKRCKENNDCCSKKCKRRGANPEKRCR

SEQ ID NO: 483 VFINVKCRGSPECLPKCKEAIGKSAGKCMNGKCKCYP

SEQ ID NO: 484 VFINAKCRGSPECLPKCKEAIGKAAGKCMNGKCKCYP

SEQ ID NO: 485 VIINVKCKISRQCLEPCKKAGMRFGKCMNGKCHCTP

SEQ ID NO: 486 VPTDVKCRGSPQCIQPCKDAGMRFGKCMNGKCHCTP

SEQ ID NO: 487 VRIPVSCKHSGQCLKPCKDAGMRFGKCMNGKCDCTP

SEQ ID NO: 488 VRIPVSCRHSGQCLRPCRDAGMRFGRCMNGRCDCTP

SEQ ID NO: 489 TNVSCTTSKECWSVCQRLHNTSRGKCMNKKCRC

SEQ ID NO: 490 NVKCTGSKQCLPACKAAVGKAAGKCMNGKCKC

SEQ ID NO: 491 GVPINVRCRGSRDCLDPCRGAGERHGRCGNSRCHCTP

SEQ ID NO: 492 VRIPVSCRHSGQCLRPCRDAGERHGRCGGGRCDCTPR

SEQ ID NO: 493 QVQTNVRCQGGSCGSVCRREGGGAGGGCGNGRCGCYRN

SEQ ID NO: 494 IKCSESYQCFPVCKSRFGKTNGRCVNGFCDCF

SEQ ID NO: 495 VKCSSPQQCLKPCKAAFGISAGgKCINGKCKCY

SEQ ID NO: 496 VSCSASSQCWPVCKKLFGTYRGKCMNSKCRCY

SEQ ID NO: 497 ESCTASNQCWSICKRLHNTNRGKCMNKKCRCY

SEQ ID NO: 498 VSCTTSKECWSVCEKLYNTSRGKCMNKKCRCY

SEQ ID NO: 499 MRCKSSKECLVKCKQATGRPNGKCMNRKCKCY

SEQ ID NO: 500 IKCTLSKDCYSPCKKETGCPRAKCINRNCKCY

SEQ ID NO: 501 IRCSGSRDCYSPCMKQTGCPNAKCINKSCKCY

SEQ ID NO: 502 IRCSGTRECYAPCQKLTGCLNAKCMNKACKCY

SEQ ID NO: 503 ISCTNPKQCYPHCKKETGYPNAKCMNRKCKCF

SEQ ID NO: 504 ASCRTPKDCADPCRKETGCPYGKCMNRKCKCN

SEQ ID NO: 505 TSCISPKQCTEPCRAKGCKHGKCMNRKCHCM

SEQ ID NO: 506 KECTGPQHCTNFCRKN-KCTHGKCMNRKCKCF

SEQ ID NO: 507 IKCRTPKDCADPCRKQTGCPHAKCMNKTCRCH

SEQ ID NO: 508 VKCTTSKECWPPCKAATGKAAGKCMNKKCKCQ

SEQ ID NO: 509 LECGASRECYDPCFKAFGRAHGKCMNNKCRCY

SEQ ID NO: 510 EKCFATSQCWTPCKKAIGSLQSKCMNGKCKCY

SEQ ID NO: 511 VRCYASRECWEPCRRVTGSAQAKCQNNQCRCY

SEQ ID NO: 512 VKCSASRECWVACKKVTGSGQGKCQNNQCRCY

SEQ ID NO: 513 VKCISSQECWIACKKVTGRFEGKCQNRQCRCY

SEQ ID NO: 514 VRCYDSRQCWIACKKVTGSTQGKCQNKQCRCY

SEQ ID NO: 515 VDCTVSKECWAPCKAAFGVDRGKCMGKKCKCY SEQ ID NO: Amino Acid Sequence

SEQ ID NO: 516 AKCRGSPECLPKCKEAIGKAAGKCMNGKCKCY

SEQ ID NO: 517 KKCQGGSCASVCRRVIGVAAGKCINGRCVCY

SEQ ID NO: 518 KKCSNTSQCYKTCEKVVGVAAGKCMNGKCICY

SEQ ID NO: 519 VKCSGSSKCVKICIDRYNTRGAKCINGRCTCY

SEQ ID NO: 520 NRCNNSSECIPHCIRIFGTRAAKCINRKCYCY

SEQ ID NO: 521 KECNGSSECYSHCEGITGKRSGKCINKKCYCY

SEQ ID NO: 522 AFCNLRRCELSCRSLGLLGKCIGEECKCV

SEQ ID NO: 523 AVCNLKRCQLSCRSLGLLGKCIGDKCECV

SEQ ID NO: 524 A AC YS S -DCR VKC V AMGFS SGKCINS KCKC Y

SEQ ID NO: 525 AICATDADCSRKCPGNPPCRNGFCACT

SEQ ID NO: 526 TECQIKNDCQRYCQSVKECKYGKCYCN

SEQ ID NO: 527 TQCQSVRDCQQYCLTPDRCSYGTCYCK

SEQ ID NO: 528 VSCRYGSDCAEPCKRLKCLLPSKCINGKCTCY

SEQ ID NO: 529 IKCRYPADCHIMCRKVTGRAEGKCMNGKCTCY

SEQ ID NO: 530 IKCSSSSSCYEPCRGVTGRAHGKCMNGRCTCY

SEQ ID NO: 531 VKCTGSKQCLPACKAAVGKAAGKCMNGKCKCY

SEQ ID NO: 532 VSCKHSGQCIKPCKDA-GMRFGKCMNRKCDCT

SEQ ID NO: 533 VKCRGSPQCIQPCRDA-GMRFGKCMNGKCHCT

SEQ ID NO: 534 VKCTSPKQCLPPCKAQFGIRAGAKCMNGKCKCY

SEQ ID NO: 535 VKCTSPKQCSKPCKELYGSSAGAKCMNGKCKCY

SEQ ID NO: 536 VKCTSPKQCLPPCKEIYGRHAGAKCMNGKCHCS

SEQ ID NO: 537 VKCTGSKQCWPVCKQMFGKPNGKCMNGKCRCY

SEQ ID NO: 538 VKCRGSRDCLDPCKKAGMRFGKCINSKCHCT

SEQ ID NO: 539 VRCVTDDDCFRKCPGNPSCKRGFCACK

SEQ ID NO: 540 VPCNNSRPCVPVCIREVNNKNGKCSNGKCLCY

SEQ ID NO: 541 VPINVKCRGSRDCLDPCKKAGMRFGKCINSKCHCTP

SEQ ID NO: 542 VQTNVKCQGGSCASVCRREIGVAAGKCINGKCVCYRN

SEQ ID NO: 543 AEIIRCSGTRECYAPCQKLTGCLNAKCMNKACKCYGCV

SEQ ID NO: 544 RPTDIKCSASYQCFPVCKSRFGKTNGRCVNGLCDCF

SEQ ID NO: 545 QFTDVKCTGSKQCWPVCKQMFGKPNGKCMNGKCRCYS

SEQ ID NO: 546 VGINVKCKHSRQCLKPCKDAGMRFGKCTNGKCHCTPK

SEQ ID NO: 547 VVIGQRCYRSPDCYSACKKLVGKATGKCTNGRCDC

SEQ ID NO: 548 NFKVEGACSKPCRKYCIDKGARNGKCINGRCHCYY

SEQ ID NO: 549 QIDTNVKCSGSSKCVKICIDRYNTRGAKCINGRCTCYP

SEQ ID NO: 550 GVPISVRCRGSRDCLEPCRRAGTRFGRCINGRCHCTP

SEQ ID NO: 551 GVPISVRCRGSRDCLEPCRRAGTRFGRCIQSRCHCTP

SEQ ID NO: 552 GVPISVRCRGSRDCLEPCRRAGTRFGRCINRRCHCTP

SEQ ID NO: 553 GVPINVRCRGSRDCLEPCRRAGTRFGRCINSRCHCTP

SEQ ID NO: 554 GVPINVRCRGSRDCLEPCRRAGTRFGRCIQSRCHCTP

SEQ ID NO: 555 GVPINVRCRGSRDCLEPCRRAGTRFGRCIQSRCHCYP

SEQ ID NO: 556 GVPINVRCRGSRDCYEPCRRAGTRFGRCIQSRCHCTP

SEQ ID NO: 557 GVPINVRCRGSRDCLEPCRRAGTRFGRCIQSRCYCTP

SEQ ID NO: 558 GVPISVRCRGSRDCLEPCRRAGTRFGRCIQSRCHCYP

SEQ ID NO: 559 GVPISVRCRGSRDCYEPCRRAGTRFGRCIQSRCHCTP

SEQ ID NO: 560 GVPISVRCRGSRDCLEPCRRAGTRFGRCIQSRCYCTP SEQ ID NO: Amino Acid Sequence

SEQ ID NO: 561 GVPINVRCRGSRDCLEPCRRAGTRFGRCIASRCHCYP

SEQ ID NO: 562 GVPINVRCRGSRDCLEPCRRAGTRFGRCISSRCHCYP

SEQ ID NO: 563 GVPINVRCRGSRDCLEPCRRAGTRFGRCITSRCHCYP

SEQ ID NO: 564 GVPINVRCRGSRDCLEPCRRAGTRFGRCINSRCHCYP

[0077] In any of SEQ ID NO: 1 - SEQ ID NO: 564 or fragment thereof, any one or more K residues can be replaced by an R residue or an A residue, any one or more R residues can be replaced by a K residue or an A residue, any one or more A residues can be replaced by a K residue or an R residue, all K residues can be replaced by R residues or A residues, all but one K residue can be replaced by R or A residues, all but two K residues can be replaced by R residues or A residues, or in any combination thereof. In any of SEQ ID NO: 1 - SEQ ID NO: 564 or any fragment thereof, any one or more M residues can be replaced by any one of I, L, or V residues, any one or more L residues can be replaced by any one of V, I, or M residues, any one or more I residues can be replaced by any one of M, L, or V residues, or any one or more V residues can be replaced by any one of I, L, or M residues. In any embodiment, at least one of the amino acids alone or in combination can be interchanged in the peptides or peptide fragments as follows: K/R, M/ II L/V, G/A, S/T, Q/N, and D/E wherein each letter is each individually any amino acid or amino acid analogue. In some instances, the peptide can contain only one lysine residue, or no lysine residue. In any of SEQ ID NO: 1 - SEQ ID NO: 564 or fragment thereof, any amino acid can be replaced with citrulline. In any of SEQ ID NO: 1 - SEQ ID NO: 564 or any fragment thereof, X can independently be any number of any amino acid or no amino acid. In some cases, a peptide can include the first two N-terminal amino acids GS, as with peptides of SEQ ID NO: 1 -SEQ ID NO: 274, or such N-terminal amino acids (GS) can be substituted by any other one or two amino acids. In other cases, a peptide does not include the first two N-terminal amino acids GS, as with peptides of SEQ ID NO: 275 - SEQ ID NO: 564. In some cases, the N-terminus of the peptide is blocked, such as by an acetyl group; in other instances the C-terminus of the peptide is block, such as by an amide group.

[0078] In some instances, the peptide is any one of SEQ ID NO: 1 -564 or a functional fragment thereof. In other embodiments, the peptide of the disclosure further comprises a peptide with 100%, 99%, 97%, 95%, 90%, 85%, or 80% homology to any one of SEQ ID NO: 1 - SEQ ID NO: 564. In further embodiments, the peptide fragment comprises a contiguous fragment of any one of SEQ ID NO: 1 - SEQ ID NO: 564 that is at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, at least 41, at least 42, at least 43, at least 44, at least 45, at least 46 residues long, wherein the peptide fragment is selected from any portion of the peptide. In some embodiments, such peptide fragments contact the cartilage and exhibit properties of those described herein for peptide and peptide- active agent conjugates.

[0079] The peptides of the present disclosure can further comprise negative amino acid residues. In some cases, the peptide has 2 or fewer negative amino acid residues. In other cases, the peptide has 4 or fewer negative amino acid residues, 3 or fewer negative amino acid residues, or 1 or fewer negative amino acid residues. The negative amino acid residues can be selected from any negative charged amino acid residues. The negative amino acid residues can selected from either E or D, or a combination of both E and D.

[0080] The peptides of the present disclosure can further comprise basic amino acid residues. In some embodiments, basic residues are added to the peptide sequence to increase the charge at physiological pH. The added basic residues can be any basic amino acid. The added basic residues can be selected from K or R, or a combination of K or R.

[0081] In some embodiments, the peptide has a charge distribution comprising an acidic region and a basic region. An acidic region can be a nub. A nub is a portion of a peptide extending out of the peptide's three-dimensional structure. A basic region can be a patch. A patch is a portion of a peptide that does not designate any specific topology characteristic of the peptide's three- dimensional structure. In further embodiments, a cystine-dense peptide can be 6 or more basic residues and 2 or fewer acidic residues.

[0082] The peptides of the present disclosure can further comprise positively charged amino acid residues. In some cases, the peptide has at least 2 positively charged residues. In other cases, the peptide has at least 3 positively charged residues, at least 4 positively charged residues, at least 5 positively charged residues, at least 6 positively charged residues, at least 7 positively charged residues, at least 8 positively charged residues or at least 9 positively charged residues. The positively charged residues can be selected from any positively charged amino acid residues. The positively charged residues can be selected from either K or R, or a combination of K and R.

[0083] In addition, the peptides herein can comprise a 4-19 amino acid residue fragment of any of the above sequences containing at least 2 cysteine residues, and at least 2 or 3 positively charged amino acid residues (for example, arginine, lysine or histidine, or any combination of arginine, lysine or histidine). In other embodiments, the peptides herein is a 20-70 amino acid residue fragment of any of the above sequences containing at least 2 cysteine residues, no more than 2 basic residues, and at least 2 or 3 positively charged amino acid residues (for example, arginine, lysine or histidine, or any combination of arginine, lysine or histidine). In some embodiments, such peptide fragments contact the cartilage and exhibit properties of those described herein for peptide and peptide- active agent conjugates. [0084] In some embodiments, the peptide contains one or more disulfide bonds and has a positive net charge at neutral pH. At physiological pH, peptides can have a net charge, for example, of -5, -4, -3, -2, -1, 0, +1, +2, +3, +4, or +5. When the net charge is zero, the peptide can be uncharged or zwitterionic. In some instances, the peptide can have a positive charge at physiological pH. In some instances, the peptide can have a charge > +2 at physiological pH, > +3. 5 at physiological pH, > +4. 5 at physiological pH. In some embodiments, the peptide contains one or more disulfide bonds and has a positive net charge at neutral pH where the net charge can be +0. 5 or less than +0. 5, +1 or less than +1, +1. 5 or less than +1. 5, +2 or less than +2, +2. 5 or less than +2. 5, +3 or less than +3, +3. 5 or less than +3. 5, +4 or less than +4, +4. 5 or less than +4. 5, +5 or less than +5, +5. 5 or less than +5. 5, +6 or less than +6, +6. 5 or less than +6. 5, +7 or less than +7, +7. 5 or less than +7. 5, +8 or less than +8, +8. 5 or less than +8. 5, +9 or less than +9. 5, +10 or less than +10. In some embodiments, the peptide has a negative net charge at physiological pH where the net charge can be -0. 5 or less than -0. 5, -1 or less than -1, -1. 5 or less than -1. 5, -2 or less than -2, -2. 5 or less than -2. 5, -3 or less than -3, -3. 5 or less than -3. 5, -4 or less than -4, -4. 5 or less than -4. 5, -5 or less than -5, -5. 5 or less than -5. 5, -6 or less than -6, -6. 5 or less than -6. 5, -7 or less than -7, -7. 5 or less than -7. 5, -8 or less than -8, -8. 5 or less than -8. 5, -9 or less than -9. 5, -10 or less than -10. In some cases, the engineering of one or more mutations within a peptide yields a peptide with an altered isoelectric point, charge, surface charge, or rheology at physiological pH. Such engineering of a mutation to a peptide derived from a scorpion or spider can change the net charge of the complex, for example, by decreasing the net charge by 1, 2, 3, 4, or 5, or by increasing the net charge by 1, 2, 3, 4, or 5. In such cases, the engineered mutation may facilitate the ability of the peptide to contact the cartilage. Suitable amino acid modifications for improving the rheology and potency of a peptide can include conservative or non-conservative mutations. A peptide can comprises at most 1 amino acid mutation, at most 2 amino acid mutations, at most 3 amino acid mutations, at most 4 amino acid mutations, at most 5 amino acid mutations, at most 6 amino acid mutations, at most 7 amino acid mutations, at most 8 amino acid mutations, at most 9 amino acid mutations, at most 10 amino acid mutations, or another suitable number as compared to the sequence of the venom or toxin that the peptide is derived from. In other cases, a peptide, or a functional fragment thereof, comprises at least 1 amino acid mutation, at least 2 amino acid mutations, at least 3 amino acid mutations, at least 4 amino acid mutations, at least 5 amino acid mutations, at least 6 amino acid mutations, at least 7 amino acid mutations, at least 8 amino acid mutations, at least 9 amino acid mutations, at least 10 amino acid mutations, or another suitable number as compared to the sequence of the venom or toxin that the peptide is derived from. In some embodiments, mutations can be engineered within a peptide to provide a peptide that has a desired charge or stability at physiological pH.

[0085] Peptides can be mutated to add function or remove function. For example, peptides and peptide-conjugates of the present disclosure can be mutated to retain, remove, or add the ability to bind to ion channels, or to promote agonizing or antagonizing ion channels, such as potassium channel binding that may occur with the peptide or peptide-conjugates (e.g., the potassium channel hERG). In some instances, it can be advantageous to remove potassium channel binding from a peptide used for delivery of an active agent. Mutations can include one or more N to S, D to E, M to T, N to Q, N to A, N to S, N to T, N to L, S to G, and S to R amino acid substitutions, or one or more L to Y, H to Y, and T to Y amino acid substitutions, or any combination of thereof, depending on whether the variant is designed to retain function or to remove function of binding to the ion channel. In some embodiments the peptides and peptide-drug conjugates of the present disclosure are mutated to minimize ion channel binding in order to minimize side effects or enhance the safety either in the target tissue or systemically.

[0086] In some embodiments, charge can play a role in cartilage homing. The interaction of a peptide of this disclosure in solution and in vivo can be influenced by the isoelectric point (pi) of the peptide and/or the pH of the solution or the local environment it is in. The charge of a peptide in solution can impact the solubility of the protein as well as parameters such as biodistribution, bioavailability, and overall pharmacokinetics. Additionally, positively charged molecules can interact with negatively charged molecules. Positively charged molecules such as the peptides disclosed herein can interact and bind with negatively charged molecules such as the negatively charged extracellular matrix molecules in the cartilage including hyaluranon and aggrecan.

Positively charged residues can also interact with specific regions of other proteins and molecules, such as negatively charged residues of receptors or electronegative regions of an ion channel pore on cell surfaces. As such, the pi of a peptide can influence whether a peptide of this disclosure can efficiently home to cartilage. Identifying a correlation between pi and cartilage homing can be an important strategy in identifying lead peptide candidates of the present disclosure. The pi of a peptide can be calculated using a number of different methods including the Expasy pi calculator and the Sillero method. The Expasy pi can be determined by calculating pKa values of amino acids as described in Bjellqvist et al., which were defined by examining polypeptide migration between pH 4.5 to pH 7.3 in an immobilized pH gradient gel environment with 9.2M and 9.8M urea at 15°C or 25°C (Bjellqvist et al. Electrophoresis. 14(10): 1023-31

(1993)). The Sillero method of calculating pi can involve the solution of a polynomial equation and the individual pKas of each amino acid. This method does not use denaturing conditions

(urea) (Sillero et al. 179(2): 319-35 (1989)) Using these pi calculation methods and quantifying the cartilage to blood ratio of peptide signal after administration to a subject can be a strategy for identifying a trend or correlation in charge and cartilage homing. In some embodiments, a peptide with a pi above biological pH (~pH 7. 4) can exhibit efficient homing to cartilage. In some embodiments, a peptide with a pi of at least 8, at least 9, at least 10, or at least 11 can efficiently home to cartilage. In other embodiments, a peptide with a pi of 11 - 12 can home most efficiently to cartilage. In certain embodiments, a peptide can have a pi of about 9. In other embodiments, a peptide can have a pi of 8 - 10. In some embodiments, more basic peptides can home more efficiently to cartilage. In other embodiments, a high pi alone may not be sufficient to cause cartilage homing of a peptide.

[0087] In some embodiments, the tertiary structure and electrostatics of a peptide of the disclosure can impact cartilage homing. Structural analysis or analysis of charge distribution can be a strategy to predict residues important in biological function, such as cartilage homing. For example, several peptides of this disclosure that home to cartilage can be grouped into a structural class defined herein as "hitchins," and can share the properties of disulfide linkages between C1-C4, C2-C5, and C3-C6. The folding topologies of peptides linked through three disulfide linkages (C1-C4, C2-C5, and C3-C6), can be broken down into structural families based on the three-dimensional arrangement of the disulfides. Some cystine-dense peptides have the

C3-C6 disulfide linkage passing through the macrocycle formed by the C1-C4 and C2-C5 disulfide linkages, hitchins have the C2-C5 disulfide linkage passing through the macrocycle formed by the C1-C4 and C3-C6 disulfide linkages, and yet other structural families have the Cl-

C4 disulfide linkage passing through the macrocycle formed by the C2-C5 and C3-C6 disulfide linkages. Variants of "hitchin" class peptides with preserved disulfide linkages at these cysteine residues, primary sequence identity, and/or structural homology can be a method of identifying or predicting other potential peptide candidates that can home to cartilage. Additionally, members and related members of the calcin family of peptides can also home to cartilage, despite having a distinct tertiary structure from the "hitchin" class of peptides. Calcin peptides are structurally a subset of the cystine-dense peptides, with cystine-dense disulfide connectivity and topology, but are further classified on the basis of functioning to bind and activate ryanodine receptors (RyRs). These receptors are calcium channels that act to regulate the influx and efflux of calcium in muscle (Schwartz et al. Br J Pharmacol 157(3):392-403. (2009)). Variants of the calcin family of peptides with preserved key residues can be one way to predict promising candidates that can home to cartilage. In some embodiments, structural analysis of a peptide of this disclosure can be determined by evaluating peptides for resistance to degradation in buffers with various proteases or reducing agents. Structural analysis of the distribution of charge density on the surface of a peptide can also be a strategy for predicting promising candidates that can home to cartilage. Peptides with large patches of positive surface charge (when at pH 7. 5) can home to cartilage.

[0088] The NMR solution structures, x-ray crystallography, or crystal structures of related structural homo logs can be used to inform mutational strategies that can improve the folding, stability, and manufacturability, while maintaining the ability of a peptide to home to cartilage. They can be used to predict the 3D pharmacophore of a group of structurally homologous scaffolds, as well as to predict possible graft regions of related proteins to create chimeras with improved properties. For example, this strategy can be used to identify critical amino acid positions and loops that can be used to design drugs with improved properties or to correct deleterious mutations that complicate folding and manufacturability for the peptides. These key amino acid positions and loops can be retained while other residues in the peptide sequences can be mutated to improve, change, remove, or otherwise modify function, homing, and activity of the peptide.

[0089] Additionally, the comparison of the primary sequences and the tertiary sequences of two or more peptides can be used to reveal sequence and 3D folding patterns that can be leveraged to improve the peptides and parse out the biological activity of these peptides. For example, comparing two different peptide scaffolds that home to cartilage can lead to the identification of conserved pharmacophores that can guide engineering strategies, such as designing variants with improved folding properties. Important pharmacophore, for example, can comprise aromatic residues or basic residues, which can be important for binding.

[0090] Improved peptides can also be engineered based upon immunogenicity information, such as immunogenicity information predicted by TEPITOPE and TEPITOPEpan. TEPITOPE is a computational approach which uses position specific scoring matrix to provide prediction rules for whether a peptide will bind to 51 different HLA-DR alleles, and TEPITOPEpan is method that uses TEPITOPE to extrapolate from HLA-DR molecules with known binding specificities to

HLA-DR molecules with unknown binding specificities based on pocket similarity. For example,

TEPITOPE and TEPITOPEpan can be used to determine immunogenicity of peptides that home to cartilage. Immunogenicity information can also be predicted using the program NetMHCII version 2. 3, which can determine the likelihood that a sequence might be presented as an immunogenic peptide via the major histocompatibility complex (MHC) presentation system of antigen presenting cells (APCs). (Nielson, M et al. BMC Bioinformatics, 8: 238 (2007); Nielsen,

M. et al. BMC Bioinformatics, 10: 296 (2009)). This program can create an immunogenicity score by predicting the binding of a peptide to MHC alleles. Strong binding alleles and weak binding alleles in each major MHC allele group (DR, DQ, and DP) can be tallied separately. The number of peptides of a specific length within the sequence (e.g., a 'core' peptide that can be nine residues long) that are immunogenic can also be tallied. Comparison of peptides or 'core' peptides with high immunogenicity to peptides or 'core' peptides with low immunogenicity can guide engineering strategies for designing variants with decreased immunogenicity. Stronger binding peptides can be more likely to generate an immune response in patient carrying that given MHC alleles. Mutating stronger binding amino acids or peptides out of a peptide sequence can reduce the immunogenicity of the entire peptide. Another aspect of immunogenicity, in addition to whether a peptide binds to a patient's MHC allele, can be whether the patient's immune cells, such as a professional antigen presenting cells such as a macrophage, a B cell, or a dendritic cell, can process the peptide. A dendritic cell can take up a protein or peptide, and then can process a peptide, such as by cleaving to form a nine residue long peptide, which then can bind to the MHC and can be presented on the surface of the dendritic cell to the immune system' s various T cells, including helper T cells and cytotoxic T cells, and thus can stimulate an immune response. The processing can involve peptide bond cleavage by enzymes and disulfide bond reduction, and thus a peptide or protein that is resistant to enzymatic cleavage and/or reduction can be resistant to processing and subsequent MHC presentation to the immune system.

Therefore, having a peptide or protein that is resistant to enzymatic cleavage and/or reduction can reduce its immunogenic potential.

[0091] Furthermore, multiple sequence alignment can also be used to inform mutational strategies using previously identified sequences, and thus providing a guide to making changes that would eliminate labile residues and immunogenic regions of a peptide sequence. Peptides can be evaluated for residues of potential biochemical instability and regions of potential immunogenicity. Then, a residue that can allow for greater peptide stability at a certain location in a peptide can be identified from a multiple sequence alignment. For example, a specific residue can be identified from a multiple sequence alignment as providing greater stability for a peptide at position previously identified as a possible risk for a significant rate of deamidation, cleavage, degradation, oxidation, hydrolysis, isomerization, disulfide exchange, racemization, beta elimination, or aggregation. This information can then be used to create peptides with greater stability or reduced immunogenicity.

[0092] In addition to utilizing co-crystal x-ray structures, NMR solution structures, and mutagenesis studies, a multiple alignment of peptide sequences can be used to identify specific amino acids or regions of high conservation that indicate an important interaction with a target or receptor (e.g., binding to a potassium channel protein) or are important for folding and structure or other properties. Once the conserved amino acid or region is identified, then amino acids replacements can be determined that maintain the important properties of the peptide, such as maintenance of the structure, reduction in immunogenicity, reduction in binding to an ion channel protein, increased stability, or any combination of thereof.

The multiple sequence alignment can also identify possible locations to add a tyrosine or tryptophan residue for spectrophoto metric reporting. Incorporation of aromatic amino acids such as Tyrosine or Tryptophan into a peptide such as SEQ ID NO: 108, which otherwise contains only amino acids of low UV absorbance at 280 nm, can be analytically advantageous. Tyrosine and Tryptophan amino acids contain aromatic ring structures. These residues have distinct absorption and emission wavelengths and good quantum yields, as shown in TABLE 2, not present in other amino acids. Both Tyrosine and Tryptophan can provide a good 'handle' for analytical detection of a peptide in solution since UV absorbance in the 250-300 nm range and peptide fluorescence is specific for these aromatic molecules. While detection of a peptide such as SEQ ID NO: 108 relies on the absorbance of the peptide bond at 220 nm, where many other materials including minor impurities in solvents also often contribute to signal, the absorbance and fluorescence properties of Tryptophan and Tyrosine containing peptides can provide for a significantly more selective and sensitive detection. Thus incorporating an aromatic amino acid can create peptides better suited for concentration and purity measurements, which can be useful during analytics, process development, manufacturing, and other drug development and drug manufacturing activities. Incorporation can be achieved either through substitutions of one or more amino acids in the peptide to Tyr and/or Trp, insertion of Tyr and/or Trp into the peptide, or via addition of Tyr and/or Trp to the N-terminus or C-terminus of the peptide.

TABLE 2. Absorbance and Fluorescence Characteristics of Tryptophan and Tyrosine.

[0093] A peptide of this disclosure can bind to chloride, potassium, or sodium channels. The peptide can also bind to calcium channels. The peptide can block potassium channels and/or sodium channels. The peptide can block calcium channels. In some embodiments, the peptide can activate any one or more of such channels. In some embodiments, the peptide can block any one or more of such channels. In some embodiments, the peptide cannot interact with any of such channels or can be mutated to reduce or remove binding to any such channels. In still other embodiments, the peptide can be a potassium channel agonist, a potassium channel antagonist, a portion of a potassium channel, a sodium channel agonist, a sodium channel antagonist, a chloride channel agonist, a chloride channel antagonist, a calcium channel agonist, a calcium channel antagonist, a hadrucalcin, a theraphotoxin, a huwentoxin, a kaliotoxin, a cobatoxin or a lectin. In some embodiments, the lectin can be SHL-Ib2. In some embodiments, the peptide can interact with, binds, inhibits, inactivates, or alters expression of ion channels or chloride channels. In some embodiments, the peptide can interact with an Navl.7 ion channel. In some embodiments, the peptide can interact with a Kv 1.3 ion channel. In still other embodiments, the peptide interacts with proteases, matrix metalloproteinase, inhibits cancer cell migration or metastases, has antimicrobial activity, or has antitumor activity. In addition to acting on matrix metalloproteinase s, the peptide can interact with other possible proteases (e.g., elastases). In some embodiments, a peptide of this disclosure can bind to multidrug resistance transporters. Peptide and peptide drug conjugate binding to and blocking multidrug resistance transporters can be used to treat bacterial infections or cancers of the joint and/or bone.

[0094] In some embodiments, the peptide has other therapeutic effects on the cartilage or structures thereof or nearby. Beta defensin expression in articular cartilage can be correlated with immunomodulatory functions as we well as osteoarthritis, autoimmune rheumatic disorders such as systemic lupus erythematosus and rheumatoid arthritis (Vordenbaumen and Schneider 2011, Varoga 2004 and Varoga 2005). In some embodiments, the peptides or their mutants inhibit beta defensins, supplement beta defensins, are competitive inhibitors of beta defensins, active or block activation of beta defensin targets, and are used as immune modulators, or to treat autoimmune, arthritis, infections, and other articular disorders.

[0095] The present disclosure can also encompass multimers of the various peptides described herein. Examples of multimers include dimers, trimers, tetramers, pentamers, hexamers, heptamers, and so on. A multimer can be a homomer formed from a plurality of identical subunits or a heteromer formed from a plurality of different subunits. In some embodiments, a peptide of the present disclosure is arranged in a multimeric structure with at least one other peptide, or two, three, four, five, six, seven, eight, nine, ten, or more other peptides. In certain embodiments, the peptides of a multimeric structure each have the same sequence. In alternative embodiments, some or all of the peptides of a multimeric structure have different sequences.

[0096] The present disclosure further includes peptide scaffolds that, e.g., can be used as a starting point for generating additional peptides. In some embodiments, these scaffolds can be derived from a variety of cystine-dense peptides. Some suitable peptides for scaffolds can include, but are not limited to, chlorotoxin, brazzein, circulin, stecrisp, hanatoxin, midkine, hefutoxin, potato carboxypeptidase inhibitor, bubble protein, attractin, oc-GI, oc-GID, μ-ΡΙΙΙΑ, ω- MVIIA, ω-CVID, χ-MrIA, p-TIA, conantokin G, contulakin G, GsMTx4, margatoxin, shK, toxin K, chymotrypsin inhibitor (CTI), and EGF epiregulin core.

[0097] In some embodiments, the peptide sequences of the disclosure are flanked by additional amino acids. One or more additional amino acids can, for example, confer a desired in vivo charge, isoelectric point, chemical conjugation site, stability, or physiologic property to a peptide.

[0098] Identifying sequence homology can be important for determining key residues that preserve cartilage homing function. For example, in some embodiments identification of conserved positively charged residues can be important in preserving cartilage homing in any homologous variants that are made. In other embodiments, identification of basic or aromatic dyads, can be important in preserving interaction and activity with Kv ion channels in

homologous variants.

[0099] Two or more peptides can share a degree of homology and share similar properties in vivo. For instance, a peptide can share a degree of homology with a peptide of the present disclosure. In some cases, a peptide of the disclosure can have up to about 20% pairwise homology, up to about 25% pairwise homology, up to about 30% pairwise homology, up to about 35% pairwise homology, up to about 40% pairwise homology, up to about 45% pairwise homology, up to about 50% pairwise homology, up to about 55% pairwise homology, up to about 60% pairwise homology, up to about 65% pairwise homology, up to about 70% pairwise homology, up to about 75% pairwise homology, up to about 80% pairwise homology, up to about 85% pairwise homology, up to about 90% pairwise homology, up to about 95% pairwise homology, up to about 96% pairwise homology, up to about 97% pairwise homology, up to about 98% pairwise homology, up to about 99% pairwise homology, up to about 99.5% pairwise homology, or up to about 99.9% pairwise homology with a second peptide. In some cases, a peptide of the disclosure can have at least about 20% pairwise homology, at least about 25% pairwise homology, at least about 30% pairwise homology, at least about 35% pairwise homology, at least about 40% pairwise homology, at least about 45% pairwise homology, at least about 50% pairwise homology, at least about 55% pairwise homology, at least about 60% pairwise homology, at least about 65% pairwise homology, at least about 70% pairwise homology, at least about 75% pairwise homology, at least about 80% pairwise homology, at least about 85% pairwise homology, at least about 90% pairwise homology, at least about 95% pairwise homology, at least about 96% pairwise homology, at least about 97% pairwise homology, at least about 98% pairwise homology, at least about 99% pairwise homology, at least about 99.5% pairwise homology, at least about 99.9% pairwise homology with a second peptide. Various methods and software programs can be used to determine the homology between two or more peptides, such as NCBI BLAST, Clustal W, MAFFT, Clustal Omega, AlignMe, Praline, or another suitable method or algorithm.

[0100] In still other instances, the variant nucleic acid molecules of a peptide of any one of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564 can be identified by either a determination of the sequence identity or homology of the encoded peptide amino acid sequence with the amino acid sequence of any one of SEQ ID NO: 24 - SEQ ID NO: 274, SEQ ID NO: 314 - SEQ ID NO: 564, or by a nucleic acid hybridization assay. Such peptide variants can include nucleic acid molecules (1) that remain hybridized with a nucleic acid molecule having the nucleotide sequence of any one of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564 (or any complement of the previous sequences) under stringent washing conditions, in which the wash stringency is equivalent to 0.5x-2xSSC with 0.1% SDS at 55-65° C, and (2) that encode a peptide having at least 70%, at least 80%, at least 90%, at least 95% or greater than 95% sequence identity or homology to the amino acid sequence of any one SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Alternatively, peptide variants of any one SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564 can be characterized as nucleic acid molecules (1) that remain hybridized with a nucleic acid molecule having the nucleotide sequence of any one SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564 (or any complement of the previous sequences) under highly stringent washing conditions, in which the wash stringency is equivalent to 0.1 x-0.

2xSSC with 0.1% SDS at 50-65°C, and (2) that encode a peptide having at least 70%, at least 80%, at least 90%, at least 95% or greater than 95% sequence identity or homology to the amino acid sequence of any one of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564.

[0101] Percent sequence identity or homology can be determined by conventional methods. See, for example, Altschul et al., Bull. Math. Bio. 48:603 (1986), and Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA 89: 10915 (1992). Briefly, two amino acid sequences are aligned to optimize the alignment scores using a gap opening penalty of 10, a gap extension penalty of 1, and the "BLOSUM62" scoring matrix of Henikoff and Henikoff (Id.). The sequence identity or homology is then calculated as: ([Total number of identical matches]/[length of the longer sequence plus the number of gaps introduced into the longer sequence in order to align the two sequences])(100).

[0102] Additionally, there are many established algorithms available to align two amino acid sequences. For example, the "FASTA" similarity search algorithm of Pearson and Lipman is a suitable protein alignment method for examining the level of sequence identity or homology shared by an amino acid sequence of a peptide disclosed herein and the amino acid sequence of a peptide variant. The FASTA algorithm is described by Pearson and Lipman, Proc. Nat'l Acad. Sci. USA 85:2444 (1988), and by Pearson, Meth. Enzymol. 183:63 (1990). Briefly, FASTA first characterizes sequence similarity by identifying regions shared by the query sequence (e.g., SEQ ID NO: 1) and a test sequence that has either the highest density of identities (if the ktup variable is 1) or pairs of identities (if ktup=2), without considering conservative amino acid substitutions, insertions, or deletions. The ten regions with the highest density of identities are then rescored by comparing the similarity of all paired amino acids using an amino acid substitution matrix, and the ends of the regions are "trimmed" to include only those residues that contribute to the highest score. If there are several regions with scores greater than the "cutoff value (calculated by a predetermined formula based upon the length of the sequence and the ktup value), then the trimmed initial regions are examined to determine whether the regions can be joined to form an approximate alignment with gaps. Finally, the highest scoring regions of the two amino acid sequences are aligned using a modification of the Needleman-Wunsch-Sellers algorithm

(Needleman and Wunsch, J. Mol. Biol. 48:444 (1970); Sellers, Siam J. Appl. Math. 26:787 (1974)), which allows for amino acid insertions and deletions. Illustrative parameters for FASTA analysis are: ktup=l, gap opening penalty=10, gap extension penalty=l, and substitution matrix=BLOSUM62. These parameters can be introduced into a FASTA program by modifying the scoring matrix file ("SMATRIX"), as explained in Appendix 2 of Pearson, Meth. Enzymol. 183:63 (1990).

[0103] FASTA can also be used to determine the sequence identity or homology of nucleic acid molecules using a ratio as disclosed above. For nucleotide sequence comparisons, the ktup value can range between one to six, preferably from three to six, most preferably three, with other parameters set as described above.

[0104] Some examples of common amino acids that are a "conservative amino acid substitution" are illustrated by a substitution among amino acids within each of the following groups: (1) glycine, alanine, valine, leucine, and isoleucine, (2) phenylalanine, tyrosine, and tryptophan, (3) serine and threonine, (4) aspartate and glutamate, (5) glutamine and asparagine, and (6) lysine, arginine and histidine. The BLOSUM62 table is an amino acid substitution matrix derived from about 2,000 local multiple alignments of protein sequence segments, representing highly conserved regions of more than 500 groups of related proteins (Henikoff and Henikoff, Proc.

Nat'l Acad. Sci. USA 89: 10915 (1992)). Accordingly, the BLOSUM62 substitution frequencies can be used to define conservative amino acid substitutions that may be introduced into the amino acid sequences of the present invention. Although it is possible to design amino acid substitutions based solely upon chemical properties (as discussed above), the language

"conservative amino acid substitution" preferably refers to a substitution represented by a BLOSUM62 value of greater than -1. For example, an amino acid substitution is conservative if the substitution is characterized by a BLOSUM62 value of 0, 1, 2, or 3. According to this system, preferred conservative amino acid substitutions are characterized by a BLOSUM62 value of at least 1 (e.g., 1, 2 or 3), while more preferred conservative amino acid substitutions are

characterized by a BLOSUM62 value of at least 2 (e.g., 2 or 3).

[0105] Determination of amino acid residues that are within regions or domains that are critical to maintaining structural integrity can be determined. Within these regions one can determine specific residues that can be more or less tolerant of change and maintain the overall tertiary structure of the molecule. Methods for analyzing sequence structure include, but are not limited to, alignment of multiple sequences with high amino acid or nucleotide identity or homology and computer analysis using available software (e.g., the Insight II. RTM. viewer and homology modeling tools; MSI, San Diego, Calif. ), secondary structure propensities, binary patterns, complementary packing and buried polar interactions (Barton, G. J. , Current Opin. Struct. Biol. 5:372-6 (1995) and Cordes, M. H. et al., Current Opin. Struct. Biol. 6:3-10 (1996)). In general, when designing modifications to molecules or identifying specific fragments determination of structure can typically be accompanied by evaluating activity of modified molecules.

[0106] Pairwise sequence alignment is used to identify regions of similarity that may indicate functional, structural and/or evolutionary relationships between two biological sequences (protein or nucleic acid). By contrast, multiple sequence alignment (MSA) is the alignment of three or more biological sequences. From the output of MSA applications, homology can be inferred and the evolutionary relationship between the sequences assessed. One of skill in the art would recognize as used herein, "sequence homology" and "sequence identity" and "percent (%) sequence identity" and "percent (%) sequence homology" have been used interchangeably to mean the sequence relatedness or variation, as appropriate, to a reference polynucleotide or amino acid sequence.

Chemical Modifications

[0107] A peptide can be chemically modified one or more of a variety of ways. In some embodiments, the peptide can be mutated to add function, delete function, or modify the in vivo behavior. One or more loops between the disulfide linkages can be modified or replaced to include active elements from other peptides (such as described in Moore and Cochran, Methods in Enzymology, 503, p. 223-251, 2012). Amino acids can also be mutated, such as to increase half-life or bioavailability, modify, add or delete binding behavior in vivo, add new targeting function, modify surface charge and hydrophobicity, or allow conjugation sites. N-methylation is one example of methylation that can occur in a peptide of the disclosure. In some embodiments, the peptide can be modified by methylation on free amines. For example, full methylation can be accomplished through the use of reductive methylation with formaldehyde and sodium

cyanoborohydride.

[0108] A chemical modification can, for instance, extend the terminal half-life, the absorption half- life, the distribution half-life of a peptide, change the biodistribution or pharmacokinetic profile, or the modification itself can be useful to provide visco supplementation to a joint. A chemical modification can comprise a polymer, a polyether, polyethylene glycol, a biopolymer, a polyamino acid, a fatty acid, a dendrimer, an Fc region, a simple saturated carbon chain such as palmitate or myristolate, sugars, hyaluronic acid, or albumin. The chemical modification of a peptide with an Fc region can be a fusion Fc-peptide. A polyamino acid can include, for example, a polyamino acid sequence with repeated single amino acids (e.g., polyglycine), and a polyamino acid sequence with mixed polyamino acid sequences (e.g., gly-ala-gly-ala (SEQ ID NO: 568)) that can or cannot follow a pattern, or any combination of the foregoing.

[0109] In some embodiments, the peptides of the present disclosure may be modified such that the modification increases the stability and/or the half-life of the peptides. In some embodiments, the attachment of a hydrophobic moiety, such as to the N-terminus, the C-terminus, or an internal amino acid, can be used to extend half-life of a peptide of the present disclosure. In other embodiments, the peptide of the present disclosure can include post-translational modifications

(e.g., methylation and/or amidation), which can affect, e.g., serum half- life. In some

embodiments, simple carbon chains (e.g., by myristoylation and/or palmitylation) can be conjugated to the peptides. In some embodiments, for example, the simple carbon chains may render conjugated peptides easily separable from unconjugated material. For example, methods that may be used to separate the desired peptides of the invention from unconjugated material include, but are not limited to, solvent extraction and reverse phase chromatography. In some embodiments, lipophilic moieties can be conjugated to the peptide and can extend half-life through reversible binding to serum albumin. Moreover, the conjugated moieties can be lipophilic moieties that extend half-life of the peptides through reversible binding to serum albumin. In some embodiments, the lipophilic moiety can be cholesterol or a cholesterol derivative including cholestenes, cholestanes, cholestadienes and oxysterols. In some

embodiments, the peptides can be conjugated to myristic acid (tetradecanoic acid) or a derivative thereof. In other embodiments, the peptides of the present disclosure are coupled (e.g., conjugated) to a half- life modifying agent. Examples of half-life modifying agents include but are not limited to: a polymer, a polyethylene glycol (PEG), a hydroxy ethyl starch, polyvinyl alcohol, a water soluble polymer, a zwitterionic water soluble polymer, a water soluble poly(amino acid), a water soluble polymer of proline, alanine and serine, a water soluble polymer containing glycine, glutamic acid, and serine, an Fc region, a fatty acid, palmitic acid, antibodies, or a molecule that binds to albumin.

[0110] In some embodiments, the first two N-terminal amino acids (GS) of SEQ ID NO: 1 - SEQ ID NO: 274 can serve as a spacer or linker in order to facilitate conjugation or fusion to another molecule, as well as to facilitate cleavage of the peptide from such conjugated or fused molecules. In some embodiments, the peptides of the present disclosure can be conjugated to other moieties that can modify or effect changes to the properties of the peptides.

Active Agent Conjugates

[0111] Peptides according to the present disclosure can be conjugated or fused to a peptide biological agent or other agent comprising amino acids (e.g., an antibody or antibody fragment, receptor or receptor fragment, ligand or ligand fragment, hormone or hormone fragment, growth factors and growth factor fragments, biological toxins and fragments thereof, or other active portion of a peptide), a protein, a peptide, or to a small molecule, RNA, DNA, or other active agent molecular structure for use in the treatment of cartilage diseases, disorders, or injuries. A peptide active agent conjugate can be a peptide conjugated to an active agent by any mechanism described herein. For example, a peptide can be covalently conjugated to an active agent to form a peptide active agent conjugate. A peptide can be chemically conjugated to an active agent to form a peptide active agent conjugate. A peptide and active agent can be expressed as a fusion protein to form a peptide active agent conjugate. For example, an antibody or fragment thereof and a peptide can be expressed as a fusion protein to form a peptide active agent conjugate. For example, in certain embodiments, a peptide as described herein can be fused to another molecule, such as an active agent that provides a functional capability. A peptide can be conjugated with an active agent through expression of a vector containing the sequence of the peptide with the sequence of the active agent. In various embodiments, the sequence of the peptide and the sequence of the active agent are expressed from the same Open Reading Frame (ORF). In various embodiments, the sequence of the peptide and the sequence of the active agent can comprise a contiguous sequence. Various vectors and recombinant systems known in the art can be employed to make such fusion peptides. The peptide and the active agent can each retain similar functional capabilities in the fusion peptide compared with their functional capabilities when expressed separately.

[0112] Furthermore, for example, in certain embodiments, the peptides described herein are attached to another molecule, such as an active agent that provides a functional capability. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 active agents can be linked to a peptide.

Multiple active agents can be attached by methods such as conjugating to multiple lysine residues and/or the N-terminus, or by linking the multiple active agents to a scaffold, such as a polymer or dendrimer and then attaching that agent-scaffold to the peptide (such as described in

Yurkovetskiy, A. V. , Cancer Res 75(16): 3365-72 (2015).

[0113] Described herein are active agents that can be conjugated to the peptides of the present invention for use in either cartilage disorders or kidney disorders, or both. In some embodiments, certain compounds or drugs are appropriate for use in either cartilage or kidney disorders, certain drug classes may be preferred for specific treatment depending on the indication or disorder. As described herein, it is understood that certain active agents are described in a non-limiting exemplary manner for use in treatments of cartilage and/or kidney indications. One or more of such active agents can be conjugated to a peptide of the present invention alone or in

combination with one or more detectable agents described herein. In some embodiments, active agents that can be conjugated to any peptide of this disclosure can be classified by mechanism.

For example, active agents can belong to the class of ant i- inflammatory drugs,

immunosuppressive (immune suppression) drugs, analgesics/pain relief drugs, disease modifying osteo arthritic drugs (DMOADs), cell depleting agents/apoptosis modifiers, bone resorptive agents and visco supplementing agents, and tissue normalization (disease modifying) drugs.

[0114] Ant i- inflammatory active agents can include, but are not limited to, corticosteroids, glucocorticoids, nonsteroidal ant i- inflammatory drugs (NSAIDs), biologies, and other small molecules. Examples of corticosteroid active agents that can be conjugated to any peptide of this disclosure for delivery to the joints and kidneys include triamcinolone dexamethasone, budesonide, and triamcinolone acetonide. Examples of NS AID active agents that can be conjugated to any peptide of this disclosure for delivery to the joints and kidneys include naproxen and ibuprofen. Other active agents include acetyls alicy lie acid and acetaminophen.

NSAID active agents can be further classified into COX2 inhibitors. An example of a COX2 inhibitor active agent directed to a prostaglandin pathway that can be conjugated to any peptide of this disclosure for delivery to the joint includes celecoxib. An example of a COX2 inhibitor active agent with anti-leukotriene receptor antagonist that can be conjugated to any peptide of this disclosure for delivery to the joint includes montelukast. An example of a COX2 inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the kidneys includes iguratimod. Biologic active agents can be further classified into active agents that are

IL-1 family inhibitors, IL-17 or IL-23 pathway inhibitors, IL-6 family inhibitors, interferon receptor inhibitors, tumor necrosis factor (TNF) inhibitors, RANK pathway inhibitors, B cell inhibitors, anti-IgE active agents, and co-stimulation inhibitors. An example of an IL-1 family inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes anakinra. An example of an IL-17/IL-23 pathway inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes secukinumab. An example of an IL-6 family inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the kidneys includes sirukumab. An example of an interferon receptor inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the kidneys includes anifrolumab. An example of a TNF inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes infliximab or etanercept. An example of a RANK pathway inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes denosumab. An example of a B cell inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the joints and kidneys includes rituximab. An example of an anti-IgE active agent that can be conjugated to any peptide of this disclosure for delivery to the kidneys includes omalizumab. An example of a co- stimulation inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes abatacept.

[0115] Pain relief active agents can include, but are not limited to analgesics, counter-irritants, and pain receptor blocking drugs. Analgesics can be further classified into non-narcotic agents and narcotic agents. An example of a non-narcotic active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes acetaminophen. An example of a narcotic active agent that can be conjugated to any peptide of this disclosure for delivery to joints includes oxycodone. Counter-irritant active agents can be further classified as natural products.

An example of a natural product that can be conjugated to any peptide of this disclosure for delivery to the joints includes capsaicin. Pain receptor blocking active agents can be further classified as TRPV4 inhibitors. An example of a TRPV4 inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes GSK2193874.

[0116] Apoptosis modifier active agents can include, but are not limited to, biologies and small molecules. Biologic apoptosis modifier active agents can be further classified as Fas/FasL inhibitors, TNF/TNFR inhibitors, TRAIL/TRAILR inhibitors, TWEAK/Fnl4 inhibitors, IL-1 inhibitors, IL-1 receptor antagonists, growth factors, and sclerostin inhibitors. An example of a

TNF/TNFR inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes infliximab. An example of a TRAIL/TRAILR inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes osteoprotegrin. An example of a TWEAK/Fnl4 inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the kidneys includes BIIB023. An example of an IL-

1 receptor antagonist that can be conjugated to any peptide of this disclosure for delivery to the joints includes anakinra. An example of a growth factor active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes IGF-1. An example of a growth factor active agent that can be conjugated to any peptide of this disclosure for delivery to the kidneys includes EGF. An example of a sclerostin inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes romosozumab. Small molecule apoptosis modifier active agents can be further classified as caspase inhibitors, iNOS inhibitors, surfactants, and bisphospho nates. An example of a caspase inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes ZVAD-fmk. An example of an iNOS inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the joints include S-methylisothiourea. An example of a surfactant active agent that can be conjugated to any peptide of this disclosure for delivery to the joints include P188. An example of a bisphosphonate active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes alendronate. Moreover, the known class of drugs called senotherapeutics, also referred to as senolytics or senolytic drugs or senolytic compounds, refers to small molecules that can selectively induce death of senescent cells and for example by directly or indirectly inducing apoptosis in senescent cells. In addition, senolytics may also act via non-apoptotic mechanisms of cell death including by necroptis, autophagic cell death, pyroptis and caspase-independent cell death (Journal of Cell Science 127; 2135-2144 (2014)). Such drugs can attenuate age-related deterioration of tissues or organs. Examples of drugs that can be conjugated to any peptide of this disclosure to induce apoptosis or induce cell death via non-apoptotic mechanisms include quercetin, dasatinib, bortezomib, carfilzomib, and navitoclax amongst other compounds disclosed herein. Additional active agents are described in the following references: Zhu, Y et al., Aging Cell 14(4):644-58 (2015); Kirkland, JL, Exp Gerontol. 48(1): 1-5 (2013); Kirkland JL and Tchkonia T, Exp Gereontol. 68: 19-25 (2015) Tchkonia, T et al., J Clin Invest., 123(3): 966-72 (2013); WO2016118859; Sugumar, D et al., Pharmagenomics Pers Med. 8: 23-33 (2015); Jiafa, R et al., Sci Rep. 6: 23968 (2016); Swanson, CD et al., Nat Rev Rheumatol., 5(6): 317-324 (2009); Oh, CJ et al., PLoS One, 7(10) :e45870 (2012); and Adebajo, A and Boehncke, W, Psoriatic Arthritis and Psoriasis: Pathology and Clinical Aspects, Springer (2016).

[0117] Tissue normalization (disease modifying) active agents can include, but are not limited to, biologies and small molecules. Biologic active agents can be further classified as chemokines (e.g., for stem cell recruitment) and growth factors. An example of a tissue normalization chemokine active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes MIP-3a. An example of a tissue normalization growth factor active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes BMP-2. Small molecule active agents can be further classified as flavonoids, ACE inhibitors, and antiproliferative active agents. An example of a tissue normalization flavonoid active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes icariin. An example of a tissue normalization ACE inhibitor active agent that can be conjugated to any peptide of this disclosure for delivery to the kidneys includes captopril. An example of a tissue normalization anti-proliferative active agent that can be conjugated to any peptide of this disclosure for delivery to the joints includes methotrexate.

[0118] TABLE 3 describes active agents for treatment of a cartilage disorder that can be conjugated to any peptide of the present disclosure to form peptide-drug conjugates.

TABLE 3 - Exemplary Active Agents for Cartilage Disorders

Active Agent Class Active Agent

Anti-Pain Drugs Duloxetine

Polymers Low Molecular Weight Chitosan

Matrix Drugs Chondroitin sulfate glucosamine

Cytokines/Growth Factors TGF-beta

Matrix Laminin

Matrix Fibronectin

Matrix Lubricin

Matrix Hyaluronic acid injections

Matrix Glucosamine

Immunosuppressants Rapamycin

HIF-Ια Modulators

HIF-2a Modulators

Corticosteroid Tixocortol pivalate

Glucocorticoid Corticosteroid Hydrocortisone Acetate

Glucocorticoid Corticosteroid Hydrocortisone t-Butyl Acetate

Glucocorticoid Corticosteroid Prednisolone Acetate

Glucocorticoid Corticosteroid Prednisolone t-Butyl Acetate

Corticosteroid Dexamethasone Acetate

Corticosteroid Dexamethasone t-Butyl Acetate

Glucocorticoid Corticosteroid Triamcinolone Diacetate

[0119] TABLE 4 describes active agents for treatment of a kidney disorder that can be conjugated to any peptide of the present disclosure to form peptide-drug conjugates.

TABLE 4 - Exemplary Active Agents for Kidney Disorders

Active Agent Class Active Agent

ACE Inhibitors Captopril

Angiotensin receptor blockers Angiotensin receptor blocker losartan (Cozaar)

Hormones Adrenocorticotropic hormone

Hormones corticotropin-releasing hormone

amphotericin B

digitalis glycosides

potassium-depleting diuretics

Coumarine anticoagulants

NLRP3 Inflammosome Targeted MCC950

Drugs

NLRP3 Inflammosome Targeted BHB

Drugs

NLRP3 Inflammosome Targeted Type I interferon

Drugs

NLRP3 Inflammosome Targeted IFN-beta

Drugs

NLRP3 Inflammosome Targeted Resveratrol

Drugs

NLRP3 Inflammosome Targeted Arglabin

Drugs

NLRP3 Inflammosome Targeted CB2R agonist

Drugs

Drugs

[0120] TABLE 5 describes active agents for treatment of a cartilage disorder and a kidney disorder that can be conjugated to any peptide of the present disclosure to form peptide-drug conjugates.

TABLE 5 - Exemplary Active Agents for Cartilage Disorders and Kidney Disorders

Active Agent Class Active Agent

p38 Inhibitors BIRB 796

p38 Inhibitors SCIO-469

p38 Inhibitors VX-702

p38 Inhibitors Pamapimod

p38 Inhibitors ARRY-797

Corticosteroids 17-monopropionate

Corticosteroids Desciclesonide

Corticosteroids Fiunisolide

Corticosteroids Mometasone furoate

Corticosteroids 22-hydroxy intermediate budesonide derivative

Corticosteroids όβ-hydroxy budesonide derivative

Corticosteroids A6-budesonide derivative

Corticosteroids 23 -hydroxy budesonide derivative

Corticosteroids 16a-butryloxyprednisolone budesonide derivative

Corticosteroids 16a-hydroxyprednisolone budesonide derivative

Corticosteroid (Beclomethasone) QVAR inhalation

Corticosteroid (Budesonide) pulmicort respules

Corticosteroid Flovent HFA 44

Corticosteroid (Mometasone) Asmanex HFA

Corticosteroid (Mometasone) Budesonide symbicort

Corticosteroid Dexamethasone sodium phosphate

Corticosteroid Tixocortol pivalate

Corticosteroid Ciclesonide

Glucocorticoids 21-nortriamcincolone acetonide

Glucocorticoids A6-triamcinolone

Glucocorticoids 6b-hydroxy triamcinolone acetonide

Glucocorticoids 21-carboxy triamcinolone acetonide

Glucocorticoids 6b-OH, 21-COOH triamcinolone acetonide

Glucocorticoids 6a fluorocortisol

Glucocorticoids 9a fluorocortisol

Glucocorticoids Al-dehydro configuration in prednisolone

Glucocorticoids 16-methylene dexamethasone derivative

Glucocorticoids 16a- methyl dexamethasone derivative

Glucocorticoids 16P-methyl betamethasone derivative

Glucocorticoids Cyclophosphamide

Glucocorticoids Mycophenolate

Glucocorticoids/Mineralocorticoids Cortisol

Glucocorticoids/Mineralocorticoids Hydrocortisone

Glucocorticoids/Mineralocorticoids Prednisolone

Glucocorticoids/Mineralocorticoids Betamethasone

Glucocorticoid Fluticasone

Glucocorticoid Fluticasone propionate

Steroid (fiunisolide) Aerobid

Steroid (fiunisolide) Aerobid-M

Steroid (fiunisolide) Aero span

Steroid (Fiunisolide) Fluticasone Furoate

Steroid (Fluticasone) Flovent HFA 110

Steroid (Fluticasone) Flovent HFA 220

Steroid (Fluticasone) Flovent Diskus 50 Active Agent Class Active Agent

Steroid (Fluticasone) Asmanex

Steroid Betamethasone acetate

Steroid Betamethasone sodium phosphate

Steroid Betamethasone valerate

Steroid Beclomethasone dipropionate

Local Anesthetic procaine hydrochloride

Local Anesthetic Novacain

Anesthetic bupivacaine hydrochloride

Anesthetic lidocaine hydrochloride

Local Anesthetic ropivacaine hydrochloride

Analgesics Morphine

Analgesics Fentanyl

Quinazo lines Feitinib/Iressa

Quinazo lines S orafenib/Nexavar

Quinazo lines Lapatinib dito sylate/Tykerb/Ty verb

Quinazo lines Sunitinib/Sutent

Quinazo lines Bortezomib/Velcade/Cytomib

Quinazo lines E verolimu s/Temsiro limu s

Quinazo lines Inhibitors of IAPS

Quinazo lines Activators of caspase pathway

Quinazo lines Activators of AKT pathway

Quinazo lines Propylpeptidase inhibitors

Quinazo lines Activators of p53

Quinazo lines Inhibitors of anti-apoptotic protein inhibitors

Prolyl Hydroxylase (PHD) Desferoxamine

Inhibitors

Prolyl Hydroxylase (PHD) Dimethyloxalylglycine (DMOG)

Inhibitors

Prolyl Hydroxylase (PHD) L-mimosine (L-mim)

Inhibitors

Aptamers Peptide aptamers

Aptamers RNA aptamer A-p50

Aptamers Peptide A aptamer TrxLeflD

Aptamers Aptamer E07

Aptamers Aptamer gemcitabine polymers

Aptamers RAGE

Aptamers Pegaptanib

Proteosome Inhibitors Bortezomib

Proteosome Inhibitors Carfilzomib

Second Generation Proteosome Ixazomib

Inhibitors

Second Generation Proteosome Delanzomib

Inhibitors

Second Generation Proteosome Oprozomib

Inhibitors

Second Generation Proteosome Marizomib

Inhibitors

Apoptosis Inhibitors FLIP agonist

Apoptosis Inhibitors nitric oxide synthase inhibitors Active Agent Class Active Agent

Apoptosis Inhibitors caspase-3 inhibitors (Z-DEVD-fmk (SEQ ID NO: 569))

Apoptosis Inhibitors caspase-9 inhibitors (Z-LEHD-fmk (SEQ ID NO: 570))

Apoptosis Inhibitors Sclerostin antagonists

Apoptosis Inhibitors/Growth Factor IGF-1

BCL-2 Agonist Apoptosis Inhibitors Oblimersen

BCL-2 Agonist Apoptosis Inhibitors Obatoclax

BCL-2 Agonist Apoptosis Inhibitors Navitoclax

BCL-2 Agonist Apoptosis Inhibitors Venetoclax (ABT-199)

BCL-2 Agonist Apoptosis Inhibitors Navotoclax (ABT-263)

BCL-2 Agonist Apoptosis Inhibitors GX01 series of compounds

BCL-2 Agonist Apoptosis Inhibitors BCL-2 small molecule antagonists

BCL-2 Agonist Apoptosis Inhibitors Tetraocarcin-A derivatives

BCL-2 Agonist Apoptosis Inhibitors Chelerythrine

BCL-2 Agonist Apoptosis Inhibitors Antimycin A derivatives

BCL-2 Agonist Apoptosis Inhibitors HA14-1

BCL-2 Agonist Apoptosis Inhibitors Synthetic compound antagonist of BH3

BCL-2 Agonist Apoptosis Inhibitors Genasense

BCL-2 Agonist Apoptosis Inhibitors ISIS 22783

BCL-2/BCL-XL Agonist Apoptosis Bispecific Antisense

Inhibitors

Proapoptotic BCL-2 Targeting Bax, Bak, Bid, Bad-derived BH3 Peptides

Drugs

Proapoptotic BCL-2 Targeting SAHBs

Drugs

Proapoptotic BCL-2 Targeting BH3Is

Drugs

BCL-2/BCL-XL Agonist Apoptosis ABT-737

Inhibitors

BCL-X Inhibitors

Apoptosis Modifiers Caspase- 1 Inhibitors

Apoptosis Modifiers Caspase- 8 Inhibitors

Pan-caspase Caspase Inhibitor IDN-6556

Pan-caspase Caspase Inhibitor IDN-6734

Pan-caspase Caspase Inhibitor VX-799

Pan-caspase Inhibitor MX1013

Pan-caspase Caspase Inhibitor M-920

Pan-caspase Caspase Activator MX-2060 derivatives

Pan-caspase Caspase Activators Small-molecule compounds

Pan-caspase Caspase Activators RGD peptides

Pan-caspase inhibitors ZVAD-fmk

Caspase- 1 ICE Inhibitors IDN-11104

Caspase- 1 ICE Inhibitors VX-756

Caspase-3 Inhibitors M-826

Caspase-3 Inhibitors M-791

Caspase-3 Inhibitors Immunocasp-3

Caspase-3 Inhibitors Ad-G/iCasp3

Caspase-3 Inhibitors PEF-F8-CP3

Caspase-6 Inhibitors Immunocasp-6

Caspase-9 Inhibitors FKBP12/caspase-9 fusion protein Active Agent Class Active Agent

IAP Antagonists BIR3 antagonists

XIAP Antagonists Capped tripeptide XIAP Antagonists

XIAP Antagonists Smac-mimetic compounds

XIAP Antagonists AEG35156/GEM®640

XIAP Inhibitors Embelin

XIAP Inhibitors XIAP antisense and RNA constructs

XIAP/cIAP-l/cIAP-2 Inhibitors Small molecule SMAC mimetics

IAP/Caspase Inhibitors HIV-Tat/polyarginine-conjugated SMAC peptides

BIR2/Caspase-3 Inhibitors TWX024

BIR2 Inhibitors Polyphenylurea derivatives

Survivin Targeting Drugs LY2181308

Survivin Targeting Drugs Ad-Survivin T34A

Anti- TWEAK Apoptosis Modifiers BIIB023

Xanthine Oxidase Inhibitors Allopurinol

Xanthine Oxidase Inhibitors Febuxostat

Xanthine Oxidase Inhibitors Zyloprin

Growth Factor bFGF

Growth Factor IGF

Growth Factor TFG-beta

Growth Factor BMP-2

Growth Factor BMP-9

Growth Factor BMP- 13

Growth Factor BMP-7

Growth Factor BMP-3 inhibitors

Growth Factor TFG-βΙ

Growth Factor OP-1

Growth Factor PDGF

Growth Factor PTH

Growth Factor PTHrP

Growth Factor MIP-3a

Growth Factor EPO

Growth Factor FGF

Growth Factor FGF-2

Growth Factor FGF- 18

Growth Factor TGF- 3

Growth Factor VEGF

Growth Factor Wnt proteins

Growth Factor EGF

Growth Factor GM-CSF

Flavonoid Icariin

Flavonoid Quercetin

Tyrosine Kinase Inhibitor (Lck/Btk Dasatinib

Inhibitor)

TRPV4 Activators GSK1016790A

TRPV4 Activators 4alpha-PDD

TRPV4 Inhibitors HC-067047

TRPV4 Inhibitors GSK2193874

NSAID Ampion

NSAID Phenylbutazone Active Agent Class Active Agent

NSAID Naproxen lysozyme conjugate

NSAID Acetal salicylic acid

DMARDs Sulfasalazine

DMARDs Leflunomide

DMARDs Hydroxychloroquine (Plaquenil)

Disease-Modifying Osteoarthritis FGF-18

Drugs (DMO AD s)

Uricosurics Sulfinpyrazone

MSC Matrix Collagen

MSC Matrix Fibrin

MSC Matrix Polylactatous

Surfactant PI 88 and other surfactants

Molecules for Bone Marrow Niches Angiopoetin

Molecules for Bone Marrow Niches Bone morphogenitic proteins

Molecules for Bone Marrow Niches Epinephrine

Molecules for Bone Marrow Niches Norepinephrine

Molecules for Bone Marrow Niches GDF5

Molecules for Bone Marrow Niches ICAN1

Molecules for Bone Marrow Niches Jagged 1

Molecules for Bone Marrow Niches Osteopontin

Molecules for Bone Marrow Niches parathyoid hormone

Molecules for Bone Marrow Niches Calcitonin

Molecules for Bone Marrow Niches steel factor

Molecules for Bone Marrow Niches Thrombopoetin

Molecules for Bone Marrow Niches vascular cell adhesion molecule 1

Chemokine Molecules for Bone CXCL12

Marrow Niches

B Cell Targeting Agents Rituximab

B Cell Targeting Agents BLys

B Cell Targeting Agents TACI

T Cell Co-stimulation Antagonists Abatacept

JAK Targeting Agents Tofacitinib

Calcineurin Inhibitors Tacrolimus

Calcineurin Inhibitors Cyclosporin

Calcineurin Inhibitors Voclosporin

COX-2 Inhibitors Iguratimod

COX-2 Inhibitors Montelukast

COX-2 Inhibitors Rofecoxib

COX-2 Inhibitors Valdecoxib

Interferon Receptor Inhibitors Anifrolumab

IFN-a Inhibitors Sifalimumab

Anti-IgE Agents Omalizumab

iNOS Inhibitors S-methylisothiourea

CD20 Antagonists/B Cell Inhibitors Ocrelizumab

BAFF Antagonists/B Cell Inhibitors Belimumab

TNF Superfamily BAFF and APRIL Atacicept

Antagonists/B cell Inhibitors

TNF-a Antagonists Thalidomide

TNF-a Antagonists Lenalidomide Active Agent Class Active Agent

TNF-a Antagonists Pomalidomide

TNF-a Antagonists Pentocifylline

TNF-a Antagonists Bupropion

TNF Antagonists Lent iviral- mediated RNAi

TNF Agonists Recombinant TNF-a

TRAIL Receptor Agonists HGS-ETR1

TRAIL Receptor Agonists HGS-ETR2

TRAIL Receptor Agonists HGS-TR2J

TRAIL Receptor Agonists PRO 1762

TRAIL Receptor Agonists TRA-8

CD95/Fas Agonists CD95-Fc

Marine Bioactive Compounds TRAIL-Resistance Overcoming Marine Bioactive

Compounds

Marine Bioactive Compounds mazamine A

Marine Bioactive Compounds marine-derived chroomycins

Marine Bioactive Compounds Carotenoids

Marine Bioactive Compounds Aplysin

Marine Bioactive Compounds Aplidin

Marine Bioactive Compounds Siphonaxanthin

Marine Bioactive Compounds pectinotoxin-2

Anti-Complement Drugs Eculizumab

PAR-2 Modulators Pepducin P2pal-18

miR-2013 Blockers Anti-sense oligonucleotides

Nrf2 Activator Dimethyl fumarate

p53 Targeting Drugs INGN201

p53 Targeting Drugs SCH58500

p53 Targeting Drugs ONYX-015

p53 Targeting Drugs C-terminal p53 peptides

p53 Targeting Drugs CDB3

p53 Targeting Drugs CP31398

p53 Targeting Drugs Prima- 1

p53 Targeting Drugs HPV E6-binding peptide aptamers

p53 Targeting Drugs Nutlins

p53 Targeting Drugs Chalcones

p53 Targeting Drugs Small peptides

p53 Targeting Drugs Pifithrin-a

p53 Targeting Drugs/ Apoptosis QP1-1002

Modifiers (T cells)

Apaf-1 Targeting Drugs/ Apoptosis QM56

Modifiers (T cells)

Apaf-1 Targeting Drugs/ Apoptosis SVTO 16426

Modifiers (T cells)

Ferro statin 16/86

BASPl Targeting Drugs/ Apoptosis BASP siRNA

Modifiers (T cells)

Anti-Inflammatory Drugs CCX140

Anti-Inflammatory Drugs CXA-10

Anti-Inflammatory Drugs/ Anti- Alkaline phosphatase

Fibrotic Drugs Active Agent Class Active Agent

Anti-Fibrotic Drugs Dnmtl inhibitors

Anti-Inflammatory Drug s/Apopto sis THR-184

Modifiers (T cells)

Immunosuppressants Lithium

β2- Adrenergic Agonists Formoterol

Anti-Inflammatory Drugs CRMD-001

Endothelin-1 Targeting Drugs Astrasentan

Vasopressin Receptor Antagonists Tolvaptan

Vasopressin Receptor Antagonists RWJ-676070

Immunosuppressants Azathioprine

Immunosuppressants Mycophenolic acid

Immunosuppressants Cyclosporine

Immune Modulators Laquinimod

Slow-acting antirheumatic drugs

(SAARDs)

Colcrys

Hormones parathyroid hormone

Hormones growth hormone

11-beta hydro xysteroid dehydrogenases

Mineralocorticoid

Proopiomelanocortin

fludrocortisonesoxycorticosterone acetate vaccines from live attenuated viruses

Aspirin

Insulin

Isonizaid

Oral hypoglycemic agents

Antacids

Carbamazepine

Cholestyramine

Colestipol

Ephedrine

Erythromycin

Mitotane

oral contraceptives

Phenobarbital

Phenytoin

Rifampin

Troleandomycin

No n- selective caspase inhibitor okadaic acid

Camptothetic

Staurosporine

HFA

Alvesco inhalation

Breo Ellipta

Advair

Mometasone

Dulera Active Agent Class Active Agent

Umeclidinium

Anoro

Reactive Oxygen Species Targeting

Drugs

Cytokines/Growth Factors TGF-beta

NOD-like receptor protein 3- dependent caspase 1 Targeting

Drugs

NSAID Etoricoxib

Apoptosis Modifiers MCL1 inhibitors

Teriparatide

BH3 mimetic s

AZD 4320

Carrier Proteins Low molecular weight human serum albumin

Ceramide Targeting Drugs

DMARDs Penicillamine

Chondrogenic factors

Anti-oxidative factors

A(1)AR agonist

S 1P(2)R antagonist

Antimalarials

BAX/BAK activating drugs

Selective GR Activators (SEGRAs)

Rapl Targeted Drugs

Seno lytic Ephrin Ligand (EFN) B 1 blockers

Seno lytic Cyclin-dependent kinase inhibitor 1A (p21)

phosphatidylinositol-4,5-bishophate 3-kinase delta catlyatic subunit (PI3KCD) blockers

Seno lytic Plasminogen-activated inhibitor-2 (PAI-2) blockers

Senesce-associated secretory

phenotype (SASP) inhibitors

Hormone Tetracosactide

[0121] TABLE 6 describes additional active agents for treatment of a cartilage disorder and a kidney disorder that can be conjugated to any peptide of the present disclosure to form peptide- drug conjugates.

TABLE 6 - Exemplary Active Agents for Cartilage Disorders and Kidney Disorders

Active Agent Class Active Agent

Peptide Oligopeptide

Peptide Polypeptide

Peptide Pep tido mimetic

Nucleic Acid Polynucleotide

Nucleic Acid Polyribonucleotide

Nucleic Acid Oligonucleotide

Nucleic Acid DNA Active Agent Class Active Agent

Nucleic Acid cDNA

Nucleic Acid ssDNA

Nucleic Acid RNA

Nucleic Acid dsRNA

Nucleic Acid micro RNA

Nucleic Acid Interfering RNA

Nucleic Acid Aptamer

Antibody single chain variable Fragment (scFv)

Antibody Antibody Fragment

Antibody Aptamer

Antibody Fc domains

Antibody Fc regions

Antibody Fc active fragments or modifications thereof

Cytokine

Cytokine antagonists Mavrilimumab

Cytokine antagonists Ixekizumab

Cytokine antagonists Tocilizumab

Cytokine antagonists Anakinra

Cytokine antagonists Ustekinumab

Cytokine antagonists Secukinumab

Interferon

Hormone

Enzymes

Growth Factor

Checkpoint Inhibitor

CD Antigen

Chemokines

Neurotransmitters

Ion Channel Inhibitors

G-protein coupled receptor

inhibitors

G-protein coupled receptor

activators

Tumor necrosis factor inhibitors

Chemical Agents

Radio sensitizers

Radioprotectants

Radionuclide

Therapeutic Small Molecules

Steroids

Corticosteroids

Ant i- inflammatory Agents Active Agent Class Active Agent

Immune Modulators Abatacept

Immune Modulators Rituximab

Complement Fixing Peptides or

Proteins

Tumor Necrosis Factor Family

Tumor Necrosis Factor (TNF) soluble receptor or antibody Inhibitors

Tumor Necrosis Factor Family

Activators

Tumor Necrosis Factor (TNF)

soluble receptor or antibody

Caspase protease inhibitors or

activators

NF-kB, RIPK1 and/or RIPK3

Inhibitors

NF-kB, RIPK1 and/or RIPK3

Activators

Death-receptor ligand activator or

inhibitor

Tumor Necrosis Factor Family

TNFR1

Agonists

Tumor Necrosis Factor Family

TNFR2

Agonists

Tumor Necrosis Factor Family

CD27/TNFRSF7

Agonists

Tumor Necrosis Factor Family

CD30/TNFRSF8

Agonists

Tumor Necrosis Factor Family

OX40/TNFRSF4

Agonists

Tumor Necrosis Factor Family

CD40/TNFRSF5

Agonists

Tumor Necrosis Factor Family

4-1BB/TNFRSF9

Agonists

Tumor Necrosis Factor Family

RANK (receptor activator of NF-kappa B/TNFRSFl 1 A) Agonists

Tumor Necrosis Factor Family

TWEAK receptor/TNFRS F 12 A

Agonists

Tumor Necrosis Factor Family

T AC 1 /TNFRS F 13 B

Agonists

Tumor Necrosis Factor Family

BAFF-R (BAFF receptor/TNFRSF13C)

Agonists

Tumor Necrosis Factor Family

HVEM (herpes virus entry mediator/TNFRSF14)

Agonists

Tumor Necrosis Factor Family

RELT/TNFRSF19L

Agonists

Tumor Necrosis Factor Family

ectodysplasin A2 isoform receptor/TNFRS 27

Agonists

Tumor Necrosis Factor Family

ectodysplasin Al

Agonists

TNF Family Member Anhidrotic Receptor Active Agent Class Active Agent

Tumor Necrosis Factor Family

Decoy Receptor 3/TNFRSF6B

Antagonists

Tumor Necrosis Factor Family

Decoy Receptor 1/TNFRSFlOC

Antagonists

Tumor Necrosis Factor Family

Decoy Receptor 2/TNFRSF10D

Antagonists

Tumor Necrosis Factor Family

DR3 (death receptor 3/TNFRSF25)

Antagonists

Tumor Necrosis Factor Family

DR4 (death receptor 4/TNFRSF10A)

Antagonists

Tumor Necrosis Factor Family

DR5 (death receptor 5/TNFRSF10B)

Antagonists

Tumor Necrosis Factor Family

DR6 (death receptor 6/TNFRSF21)

Antagonists

Tumor Necrosis Factor Family

Fas/TNFRSF6

Antagonists

Tumor Necrosis Factor Family

Lympho toxin b receptor/TNFRS3

Antagonists

Tumor Necrosis Factor Family

OPG (osteoprotegerin/TNFRSFl lB)

Antagonists

Tumor Necrosis Factor Family

Nerve Growth Factor Receptor/TNFRSF16

Antagonists

Tumor Necrosis Factor Family

BCMA (B Cell Maturation Antigen/TNFRSF17)

Antagonists

Tumor Necrosis Factor Family

GITR (Glucocorticoid-Induced TNF Receptor/TNFRSFl 8) Antagonists

Tumor Necrosis Factor Family

TAJ (Toxicity and JNK Inducer/TNFRS F 19)

Antagonists

Tumor Necrosis Factor Family

TNFRSF22

Antagonists

Tumor Necrosis Factor Family

TNFRSF23

Antagonists

TNF Receptor Superfamily Ligands TNF alpha

TNF Receptor Superfamily Ligands Lymphotoxin-a

TNF Receptor Superfamily Ligands Tumor Necrosis Factor Membrane Form

TNF Receptor Superfamily Ligands Tumor Necrosis Factor Shed Form

TNF Receptor Superfamily Ligands LIGHT

TNF Receptor Superfamily Ligands Lympho toxin b2al heterotrimer

TNF Receptor Superfamily Ligands OX-40 Ligand

TNF Receptor Superfamily Ligands Compound 1 [PMID: 24930776]

TNF Receptor Superfamily Ligands CD40 Ligand

TNF Receptor Superfamily Ligands Fas Ligand

TNF Receptor Superfamily Ligands TL1A

TNF Receptor Superfamily Ligands CD70

TNF Receptor Superfamily Ligands CD30 Ligand

TNF Receptor Superfamily Ligands TRAF1

TNF Receptor Superfamily Ligands TRAF2 Active Agent Class Active Agent

TNF Receptor Superfamily Ligands TRAF3

TNF Receptor Superfamily Ligands TRAIL

TNF Receptor Superfamily Ligands RANK Ligand

TNF Receptor Superfamily Ligands APRIL

TNF Receptor Superfamily Ligands BAFF

TNF Receptor Superfamily Ligands B and T lymphocyte Attenuators

TNF Receptor Superfamily Ligands NGF

TNF Receptor Superfamily Ligands BDNF

TNF Receptor Superfamily Ligands Neurotrophin-3

TNF Receptor Superfamily Ligands Neurotrophin-4

TNF Receptor Superfamily Ligands TL6

TNF Receptor Superfamily Ligands Ectodysplasin A2

TNF Receptor Superfamily Ligands Ectodysplasin Al

TNF blockers Remicade (infliximab)

TNF blockers Enbrel (etanercept)

TNF blockers Humira (adalimumab)

TNF blockers Cimzia (certolizumab pegol)

TNF blockers Simponi (golimumab)

Tumor Necrosis Factor Receptor

Family Agonists

Toll Like Receptors Agonist

TIMP-3 Inhibitors

BCL-2 Family Inhibitors

IAP Disruptors

Protease Inhibitors

Amino Sugars

Chemotherapeutic

Cytotoxic chemical

Toxins

Tyrosine Kinase inhibitors Imatinib Mesylate

Protons

Antivascular Agents Bevacizumab

EGFR Inhibitors Erlotinib

Anti- Infective Agents

Antibiotics

Anti- Viral Agents

Anti-Fungal Agents

Aminoglycoside

Statins

Nanoparticles

Liposomes

Polymers Biopolymers Active Agent Class Active Agent

Polysaccharide

Proteoglycan

Glycosaminoglycans

Polyethylene glycol

Lipids

Dendrimers

Fatty Acids

Glucocorticoid

Corticosteroid

Collagenase Inhibitor

Matrix Metalloprotease Inhibitors MMP-13 inhibitor

Vitamins Vitamin D

Antibiotics

Antiviral

Antifungal

Statins

Immune Modulators

Radioisotopes

Toxins

Enzymes

Sensitizing drugs

Anti- Angiogenic Agents Cisplatin

Anti- Angiogenic Agents Anti-Metabolites

Anti- Angiogenic Agents Mitotic Inhibitors

Anti- Angiogenic Agents Growth Factor Inhibitors

Chemotherapeutic Agent Paclitaxel

Chemotherapeutic Agent Temozolomide

Chemotherapeutic Agent Topotecan

Chemotherapeutic Agent Fluorouracil

Chemotherapeutic Agent Vincristine

Chemotherapeutic Agent Vinblastine

Chemotherapeutic Agent Procarbazine

Chemotherapeutic Agent Decarbazine

Chemotherapeutic Agent Altretamine

Chemotherapeutic Agent Methotrexate

Chemotherapeutic Agent Mercaptopurine

Chemotherapeutic Agent Thioguanine

Chemotherapeutic Agent Fludarabine Phosphate

Chemotherapeutic Agent Cladribine

Chemotherapeutic Agent Pento statin

Chemotherapeutic Agent Cytarabine Active Agent Class Active Agent

Chemotherapeutic Agent Azacitidine

Chemotherapeutic Agent Etoposide

Chemotherapeutic Agent Teniposide

Chemotherapeutic Agent Irinotecan

Chemotherapeutic Agent Docetaxel

Chemotherapeutic Agent Doxorubicin

Chemotherapeutic Agent Daunorubicin

Chemotherapeutic Agent Dactinomycin

Chemotherapeutic Agent Idarubicin

Chemotherapeutic Agent Plicamycin

Chemotherapeutic Agent Mitomycin

Chemotherapeutic Agent Bleomycin

Chemotherapeutic Agent Tamoxifen

Chemotherapeutic Agent Flutamide

Chemotherapeutic Agent Leuprolide

Chemotherapeutic Agent Goserelin

Chemotherapeutic Agent Aminogluthimide

Chemotherapeutic Agent Anastrozole

Chemotherapeutic Agent Amsacrine

Chemotherapeutic Agent Asparaginase

Chemotherapeutic Agent Mitoxantrone

Chemotherapeutic Agent Mitotane

Chemotherapeutic Agent Amifostine

Apoptotic Agents

Cell Death or Cell Killing Agents Caspases

Apoptosis Activators

Apoptosis Inhibitors XBP-1

Apoptosis Inhibitors Bcl-2

Apoptosis Inhibitors Bcl-Xl

Apoptosis Inhibitors Bcl-w

Nonsteroidal Anti- Inflammatory

COX-2 Inhibitors Drugs (NSAID)

Nonsteroidal Anti- Inflammatory

Ketorolac

Drugs (NSAID)

Nonsteroidal Anti- Inflammatory

Indomethacin

Drugs (NSAID)

Nonsteroidal Anti- Inflammatory

Etodolac

Drugs (NSAID)

Nonsteroidal Anti- Inflammatory

Tolemetin

Drugs (NSAID)

Nonsteroidal Anti- Inflammatory

Naproxen

Drugs (NSAID)

Nonsteroidal Anti- Inflammatory

Enolic Acid Derivatives Drugs (NSAID) Active Agent Class Active Agent

Nonsteroidal Anti- Inflammatory

Anthranilic Acid Derivatives

Drugs (NSAID)

Nonsteroidal Anti- Inflammatory

Celecoxib

Drugs (NSAID)

Nonsteroidal Anti- Inflammatory

Sulfonanilides

Drugs (NSAID)

Nonsteroidal Anti- Inflammatory

Salicylates

Drugs (NSAID)

Nonsteroidal Anti- Inflammatory

Aceclofenac

Drugs (NSAID)

Nonsteroidal Anti- Inflammatory

Nabumetone

Drugs (NSAID)

Nonsteroidal Anti- Inflammatory

Sulindac

Drugs (NSAID)

Nonsteroidal Anti- Inflammatory

Diclofenac

Drugs (NSAID)

Nonsteroidal Anti- Inflammatory

Ibuprofen

Drugs (NSAID)

Steroids Dexamethasone

Steroids Budesonide

Steroids Triamcinolone

Steroids Triamcinolone acetonide

Steroids Cortisone

Steroids Prednisone

Steroids Prednisolone

Steroids Triamcinolone Hexacetonide

Steroids Methylprednisolone

Pain Reliever Acetaminophen

Opioids

Local Anesthetics

Anti-Depressants

Glutamate Receptor Antagonists

Adenosine

Neuropeptides

Uricase

Elastase

[0122] Further examples of active agents include but are not limited to: a peptide, an oligopeptide, a polypeptide, a peptido mimetic, a polynucleotide, a polyribonucleotide, a DNA, a cDNA, a ssDNA, a RNA, a dsRNA, a micro RNA, an RNAi, an oligonucleotide, an antibody, a single chain variable fragment (scFv), an antibody fragment, an aptamer, a cytokine, an interferon, a hormone, an enzyme, a growth factor, a checkpoint inhibitor, a PD-1 inhibitor, a PD-Ll inhibitor, a CTLA4 inhibitor, a CD antigen, aa chemokine, a neurotransmitter, an ion channel inhibitor, a G-protein coupled receptor inhibitor, a G-protein coupled receptor activator, a chemical agent, a radio sensitizer, a radioprotectant, a radionuclide, a therapeutic small molecule, a steroid, a corticosteroid, an anti- inflammatory agent, an immune modulator, a complement fixing peptide or protein, a tumor necrosis factor inhibitor, a tumor necrosis factor activator, a tumor necrosis factor receptor family agonist, a tumor necrosis receptor antagonist, a tumor necrosis factor (TNF) soluble receptor or antibody, caspase protease activator or inhibitor, an NF-KB a RIPK1 and/or RIPK3 inhibitor or activator (e.g., through To 11- like receptors (TLRs)

TLR-3 and/or TLR-4, or T-cell receptor (TCR) and the like), a death-receptor ligand (e.g., Fas ligand) activator or inhibitor, TNF receptor family (e.g., TNFR1, TNFR2, lymphotoxin β receptor/TNFRS3, OX40/TNFRSF4, CD40/TNFRSF5, Fas/TNFRSF6, decoy receptor

3/TNFRSF6B, CD27/TNFRSF7, CD30/TNFRSF8, 4- 1BB/TNFRSF9, DR4 (death receptor

4/TNFRS 10A), DR5 (death receptor 5/TNFRSF10B), decoy receptor 1/TNFRSFlOC, decoy receptor 2/TNFRSF10D, RANK (receptor activator of NF-kappa B/TNFRSF11A), OPG

(osteoprotegerin/TNFRSFl lB), DR3 (death receptor 3/TNFRSF25), TWEAK

receptor/TNFRS F 12 A, TAC1/TNFRSF13B, BAFF-R (BAFF receptor/TNFRSF13C), HVEM

(herpes virus entry mediator/TNFRSF14), nerve growth factor receptor/TNFRSF16, BCMA (B cell maturation antigen/TNFRSF17), GITR (glucocorticoid-induced TNF receptor/TNFRSF18),

TAJ (toxicity and JNK inducer/TNFRSF19), RELT/TNFRS F 19L, DR6 (death receptor

6/TNFRSF21), TNFRSF22, TNFRSF23, ectodysplasin A2 iso form receptor/TNFRS 27, ectodysplasin 1, and anhidrotic receptor, a TNF receptor superfamily ligand including - TNF alpha, lymphotoxin-a, tumor necrosis factor membrane form, tumor necrosis factor shed form,

LIGHT, lymphotoxin β ₂ αι heterotrimer, OX-40 ligand, compound 1 [PMID: 24930776], CD40 ligand, Fas ligand, TLIA, CD70, CD30 ligand, TRAFl, TRAF2, TRAF3, TRAIL, RANK ligand,

APRIL, BAFF, B and T lymphocyte attenuator, NGF, BDNF, neurotrophin-3, neurotrophin-4,

TL6, ectodysplasin A2, ectodysplasin Al - a TIMP-3 inhibitor, a BCL-2 family inhibitor, navitoclax (Aging Cell. 15(3): 428-435. (2016)) an IAP disruptor, a protease inhibitor, an amino sugar, a chemotherapeutic (whether acting through an apoptotic or non-apoptotic pathway) (Ricci et al. Oncologist l l(4):342-57 (2006)), a cytotoxic chemical, a toxin, a tyrosine kinase inhibitor

(e.g., imatinib mesylate), protons, bevacuzimab (antivascular agent), erlotinib (EGFR inhibitor), an anti- infective agent, an antibiotic, an anti- viral agent, an anti-fungal agent, an aminoglycoside, a nonsteroidal anti- inflammatory drug (NSAID), a statin, a nanoparticle, a liposome, a polymer, a biopolymer, a polysaccharide, a proteoglycan, a glycosaminoglycan, polyethylene glycol, a lipid, a dendrimer, a fatty acid, or an Fc domain or an Fc region, or an active fragment or a

modification thereof. Any combination of the above active agents can be co-delivered with peptides or peptide conjugates of this disclosure. Additionally, in some embodiments, other co- therapies such as proton therapy or ablative radiotherapy can be administered to a subject in need thereof along with peptides or peptide conjugates of this disclosure. In some embodiments, the peptide is covalently or non-covalently linked to an active agent, e.g., directly or via a linker. TNF blockers suppress the immune system by blocking the activity of TNF, a substance in the body that can cause inflammation and lead to immune-system diseases, such as Crohn's disease, ulcerative colitis, rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis and plaque psoriasis. The drugs in this class include Remicade (infliximab), Enbrel (etanercept), Humira (adalimumab), Cimzia (certolizumab pegol) and Simponi (golimumab). The peptide disclosed herein can be used to home, distribute to, target, directed to, is retained by, accumulate in, migrate to, and/or bind to cartilage, and thus also be used for localizing the attached or fused active agent. Furthermore, cystine-dense chlorotoxin peptide can be internalized in cells

(Wiranowska, M. , Cancer Cell Int. , 11: 27 (2011)). Therefore, cellular internalization, subcellular localization, and intracellular trafficking after internalization of the peptide itself, or an active agent peptide conjugate or fusion peptide can be important factors in the efficacy of an active agent conjugate or fusion. (Ducry, L., Antibody Drug Conjugates (2013); and Singh, S. K., Pharm Res., 32(11): 3541-3571 (2015)). Exemplary linkers suitable for use with the

embodiments herein are discussed in further detail below.

[0123] The peptides or peptide- active agent fusions of the present disclosure can also be conjugated to other moieties that can serve other roles, such as providing an affinity handle (e.g., biotin) for retrieval of the peptides from tissues or fluids. For example, peptides or peptide- active agent fusions of the present disclosure can also be conjugated to biotin. In addition to extension of half-life, biotin could also act as an affinity handle for retrieval of peptides or peptide- active agent fusions from tissues or other locations. In some embodiments, fluorescent biotin conjugates that can act both as a detectable label and an affinity handle can be used. Non limiting examples of commercially available fluorescent biotin conjugates include Atto 425-Biotin, Atto 488-

Biotin, Atto 520-Biotin, Atto-550 Biotin, Atto 565-Biotin, Atto 590-Biotin, Atto 610-Biotin, Atto

620-Biotin, Atto 655-Biotin, Atto 680-Biotin, Atto 700-Biotin, Atto 725-Biotin, Atto 740-Biotin, fluorescein biotin, biotin-4-fluorescein, biotin-(5-fluorescein) conjugate, and biotin-B- phycoerythrin, Alexa fluor 488 biocytin, Alexa flour 546, Alexa Fluor 549, lucifer yellow cadaverine biotin-X, Lucifer yellow biocytin, Oregon green 488 biocytin, biotin-rhodamine and tetramethylrhodamine biocytin. In some other examples, the conjugates could include

chemiluminescent compounds, colloidal metals, luminescent compounds, enzymes,

radioisotopes, and paramagnetic labels. In some embodiments, the peptide- active agent fusions described herein can be attached to another molecule. For example, the peptide sequence also can be attached to another active agent (e.g., small molecule, peptide, polypeptide, polynucleotide, antibody, aptamer, cytokine, growth factor, neurotransmitter, an active fragment or modification of any of the preceding, fluorophore, radioisotope, radionuclide chelator, acyl adduct, chemical linker, or sugar, etc.). In some embodiments, the peptide can be fused with, or covalently or non- covalently linked to an active agent.

[0124] Additionally, more than one peptide sequence can be present on or fused with a particular peptide. A peptide can be incorporated into a biomolecule by various techniques, for example by a chemical transformation, such as the formation of a covalent bond, such as an amide bond, or by solid phase or solution phase peptide synthesis, or by preparing a nucleic acid sequence encoding the biomolecule, wherein the nucleic acid sequence includes a subsequence that encodes the peptide. The subsequence can be in addition to the sequence that encodes the biomolecule, or can substitute for a subsequence of the sequence that encodes the biomolecule.

Detectable Agent Conjugates

[0125] Described herein are agents that can be conjugated to the peptides of the present invention for use in detection and tracing either cartilage disorders or kidney disorders, or both. As described herein, it is understood that certain active agents are described in a non-limiting exemplary manner for use in diagnostics, aiding surgery and treatment, prognosis and tracking of progress or remission of cartilage and/or kidney disorders, diseases or injury. One or more of such detectable agents can be conjugated to a peptide of the present invention alone or in combination with one or more active agents described herein. Moreover some detectable agents (e.g., radionuclides, radioisotopes, radio sensitizers and photo sensitizers amongst others) may also exert therapeutic activity as well. A peptide can be conjugated to an agent used in imaging, research, therapeutics, theranostics, pharmaceuticals, chemotherapy, chelation therapy, targeted drug delivery, and radiotherapy. The agent can be a detectable agent. In some embodiments, a peptide of the present invention is conjugated to detectable agents, such as a metal, a

radioisotope, a dye, fluorophore, or another suitable material that can be used in imaging. Non- limiting examples of radioisotopes include alpha emitters, beta emitters, positron emitters, and gamma emitters. In some embodiments, the metal or radioisotope is selected from the group consisting of actinium, americium, bismuth, cadmium, cesium, cobalt, europium, gadolinium, iridium, lead, lutetium, manganese, palladium, polonium, radium, ruthenium, samarium, strontium, technetium, thallium, and yttrium. In some embodiments, the metal is actinium, bismuth, lead, radium, strontium, samarium, or yttrium. In some embodiments, the radioisotope is actinium- 225 or lead-212. In some embodiments, the fluorophore is a fluorescent agent emitting electromagnetic radiation at a wavelength between 650 nm and 4000 nm, such emissions being used to detect such agent. In some embodiments the fluorophore is a fluorescent agent is selected from the group consisting of non-limiting examples of fluorescent dyes that could be used as a conjugating molecule (or as applied to each class of molecules) in the present disclosure include DyLight-680, DyLight-750, VivoTag-750, DyLight-800, IRDye-800,

VivoTag-680, Cy5. 5, or indocyanine green (ICG class of dyes). In some embodiments, near infrared dyes include cyanine dyes. Additional non-limiting examples of fluorescent dyes for use as a conjugating molecule in the present disclosure include acradine orange or yellow, Alexa Fluors and any derivative thereof, 7-actinomycin D, 8-anilinonaphthalene-l-sulfonic acid, ATTO dye and any derivative thereof, auramine-rhodamine stain and any derivative thereof,

bensantrhone, bimane, 9-10-bis(phenylethynyl)anthracene, 5,12 - bis(phenylethynyl)naththacene, bisbenzimide, brainbow, calcein, carbodyfluorescein and any derivative thereof, l-chloro-9,10-bis(phenylethynyl)anthracene and any derivative thereof, DAPI,

DiOC6, DyLight Fluors and any derivative thereof, epicocconone, ethidium bromide, FlAsH-

EDT2, Fluo dye and any derivative thereof, FluoProbe and any derivative thereof, Fluorescein and any derivative thereof, Fura and any derivative thereof, GelGreen and any derivative thereof,

GelRed and any derivative thereof, fluorescent proteins and any derivative thereof, m isoform proteins and any derivative thereof such as for example mCherry, hetamethine dye and any derivative thereof, hoeschst stain, iminocoumarin, indian yellow, indo-1 and any derivative thereof, laurdan, lucifer yellow and any derivative thereof, luciferin and any derivative thereof, luciferase and any derivative thereof, mercocyanine and any derivative thereof, nile dyes and any derivative thereof, perylene, phloxine, phyco dye and any derivative thereof, propium iodide, pyranine, rhodamine and any derivative thereof, ribogreen, RoGFP, rubrene, stilbene and any derivative thereof, sulforhodamine and any derivative thereof, SYBR and any derivative thereof, synapto-pHluorin, tetraphenyl butadiene, tetrasodium tris, Texas Red, Titan Yellow, TSQ, umbelliferone, violanthrone, yellow fluorescent protein and YOYO-1. Other Suitable fluorescent dyes include, but are not limited to, fluorescein and fluorescein dyes (e.g., fluorescein

isothiocyanine or FITC, naphthofluorescein, 4', 5'-dichloro-2',7' -dimethoxyfluorescein, 6- carboxyfluorescein or FAM, etc.), carbocyanine, merocyanine, styryl dyes, oxonol dyes, phycoerythrin, erythrosin, eosin, rhodamine dyes (e.g., carboxytetramethyl-rhodamine or

TAMRA, carboxyrhodamine 6G, carboxy-X-rhodamine (ROX), lissamine rhodamine B, rhodamine 6G, rhodamine Green, rhodamine Red, tetramethylrhodamine (TMR), etc.), coumarin and coumarin dyes (e.g., methoxycoumarin, dialkylaminocoumarin, hydroxycoumarin, aminomethylcoumarin (AMCA), etc.), Oregon Green Dyes (e.g., Oregon Green 488, Oregon

Green 500, Oregon Green 514, etc.), Texas Red, Texas Red-X, SPECTRUM RED, SPECTRUM

GREEN, cyanine dyes (e.g., CY-3, Cy-5, CY-3. 5, CY-5. 5, etc.), ALEXA FLUOR dyes (e.g.,

ALEXA FLUOR 350, ALEXA FLUOR 488, ALEXA FLUOR 532, ALEXA FLUOR 546, ALEXA FLUOR 568, ALEXA FLUOR 594, ALEXA FLUOR 633, ALEXA FLUOR 660, ALEXA FLUOR 680, etc.), BODIPY dyes (e.g., BODIPY FL, BODIPY R6G, BODIPY TMR, BODIPY TR, BODIPY 530/550, BODIPY 558/568, BODIPY 564/570, BODIPY 576/589, BODIPY 581/591, BODIPY 630/650, BODIPY 650/665, etc.), IRDyes (e.g., IRD40, IRD 700, IRD 800, etc.), indocyanine green dyes and the like. For each of the above listed fluorescent dyes various activated forms can be used for conjugation. Additional suitable detectable agents are described in PCT/US 14/56177. Non-limiting examples of radioisotopes include alpha emitters, beta emitters, positron emitters, and gamma emitters. In some embodiments, the metal or radioisotope is selected from the group consisting of actinium, americium, bismuth, cadmium, cesium, cobalt, europium, gadolinium, iridium, lead, lutetium, manganese, palladium, polonium, radium, ruthenium, samarium, strontium, technetium, thallium, and yttrium. In some

embodiments, the metal is actinium, bismuth, lead, radium, strontium, samarium, or yttrium. In some embodiments, the radioisotope is actinium-225 or lead-212.

[0126] Other embodiments of the present disclosure provide peptides conjugated to a

radio sensitizer or photo sensitizer. Examples of radio sensitizers include but are not limited to: ABT-263, ABT-199, WEHI-539, paclitaxel, carboplatin, cisplatin, oxaliplatin, gemcitabine, etanidazole, misonidazole, tirapazamine, and nucleic acid base derivatives (e.g., halogenated purines or pyrimidines, such as 5-fluorodeoxyuridine). Examples of photo sensitizers include but are not limited to: fluorescent molecules or beads that generate heat when illuminated, porphyrins and porphyrin derivatives (e.g., chlorins, bacteriochlorins, isobacteriochlorins, phthalocyanines, and naphthalocyanines), metalloporphyrins, metallophthalocyanines, angelicins,

chalcogenapyrrillium dyes, chlorophylls, coumarins, flavins and related compounds such as alloxazine and riboflavin, fullerenes, pheophorbides, pyropheophorbides, cyanines (e.g., merocyanine 540), pheophytins, sapphyrins, texaphyrins, purpurins, porphycenes,

phenothiaziniums, methylene blue derivatives, naphthalimides, nile blue derivatives, quinones, perylenequinones (e.g., hypericins, hypocrellins, and cercosporins), psoralens, quinones, retinoids, rhodamines, thiophenes, verdins, xanthene dyes (e.g., eosins, erythrosins, rose bengals), dimeric and oligomeric forms of porphyrins, and prodrugs such as 5-aminolevulinic acid. Advantageously, this approach allows for highly specific targeting of diseased cells (e.g., cancer cells) using both a therapeutic agent (e.g., drug) and electromagnetic energy (e.g., radiation or light) concurrently. In some embodiments, the peptide is covalently or non- covalently linked to the agent, e.g., directly or via a linker. Exemplary linkers suitable for use with the embodiments herein are discussed in further detail below. Linkers

[0127] Peptides according to the present disclosure that home, target, migrate to, are retained by, accumulate in, and/or bind to, or are directed to the cartilage can be attached to another moiety (e.g., an active agent), such as a small molecule, a second peptide, a protein, an antibody, an antibody fragment, an aptamer, polypeptide, polynucleotide, a fluorophore, a radioisotope, a radionuclide chelator, a polymer, a biopolymer, a fatty acid, an acyl adduct, a chemical linker, or sugar or other active agent described herein through a linker, or directly in the absence of a linker.

[0128] A peptide can be directly attached to another molecule by a covalent attachment. For example, the peptide is attached to a terminus of the amino acid sequence of a larger polypeptide or peptide molecule, or is attached to a side chain, such as the side chain of a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, a non-natural amino acid residue, or glutamic acid residue. The attachment can be via an amide bond, an ester bond, an ether bond, a carbamate bond, a carbon-nitrogen bond, a triazole, a macrocycle, an oxime bond, a hydrazone bond, a carbon-carbon single double or triple bond, a disulfide bond, or a thioether bond. In some embodiments, similar regions of the disclosed peptide(s) itself (such as a terminus of the amino acid sequence, an amino acid side chain, such as the side chain of a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, a non-natural amino acid residue, or glutamic acid residue, via an amide bond, an ester bond, an ether bond, a carbamate bond, a carbon-nitrogen bond, a triazole, a macrocycle, an oxime bond, a hydrazone bond, a carbon-carbon single double or triple bond, a disulfide bond, or a thioether bond, or linker as described herein) can be used to link other molecules.

[0129] Attachment via a linker can involve incorporation of a linker moiety between the other molecule and the peptide. The peptide and the other molecule can both be covalently attached to the linker. The linker can be cleavable, labile, non-cleavable, stable self- immolating, hydrophilic, or hydrophobic. As used herein, the term "non-cleavable" (such as used in association with an amide, cyclic, or carbamate linker or as otherwise as described herein) is often used by a skilled artisan to distinguish a relatively stable structure from one that is more labile or "cleavable" (e.g., as used in association with cleavable linkers that may be dissociated or cleaved structurally by enzymes, proteases, self-immolation, pH, reduction, hydrolysis, certain physiologic conditions, or as otherwise described herein). It is understood that "non-cleavable" linkers offer stability against cleavage or other dissociation as compared to "cleavable" linkers, and the term is not intended to be considered an absolute non-cleavable or non-dissociative structure under any conditions. Consequently, as used herein, a "non-cleavable" linker is also referred to as a "stable" linker. The linker can have at least two functional groups with one bonded to the peptide, the other bonded to the other molecule, and a linking portion between the two functional groups.

[0130] Non-limiting examples of the functional groups for attachment can include functional groups capable of forming an amide bond, an ester bond, an ether bond, a carbonate bond, a carbamate bond, or a thioether bond. Non-limiting examples of functional groups capable of forming such bonds can include amino groups; carboxyl groups; hydroxyl groups; aldehyde groups; azide groups; alkyne and alkene groups; ketones; hydrazides; acid halides such as acid fluorides, chlorides, bromides, and iodides; acid anhydrides, including symmetrical, mixed, and cyclic anhydrides; carbonates; carbonyl functionalities bonded to leaving groups such as cyano, succinimidyl, and N-hydroxysuccinimidyl; hydroxyl groups; sulfhydryl groups; and molecules possessing, for example, alkyl, alkenyl, alkynyl, allylic, or benzylic leaving groups, such as halides, mesylates, tosylates, triflates, epoxides, phosphate esters, sulfate esters, and besylates.

[0131] Non-limiting examples of the linking portion can include alkylene, alkenylene, alkynylene, polyether, such as polyethylene glycol (PEG), hydroxy carboxylic acids, polyester, polyamide, polyamino acids, polypeptides, cleavable peptides, valine-citrulline,

aminobenzylcarbamates, D-amino acids, and polyamine, any of which being unsubstituted or substituted with any number of substituents, such as halogens, hydroxyl groups, sulfhydryl groups, amino groups, nitro groups, nitroso groups, cyano groups, azido groups, sulfoxide groups, sulfone groups, sulfonamide groups, carboxyl groups, carboxaldehyde groups, imine groups, alkyl groups, halo-alkyl groups, alkenyl groups, halo-alkenyl groups, alkynyl groups, halo-alkynyl groups, alkoxy groups, aryl groups, aryloxy groups, aralkyl groups, arylalkoxy groups, heterocyclyl groups, acyl groups, acyloxy groups, carbamate groups, amide groups, urethane groups, epoxides, and ester groups.

[0132] A peptide and drug conjugated via a linker is described with the formula Peptide- A-B-C-

Drug, wherein the linker is A-B-C. A can be a stable amide link, is an amine on the peptide and the linker and can be achieved via a tetrafluorophenyl (TFP) ester or an NHS ester. B can be (-

CH2-) _X - or a short PEG (-CH ₂ CH ₂ 0-) _x (x is 1-10), and C can be the ester bond to the hydroxyl or carboxylic acid on the drug. In some embodiments, C can refer to the "cleavable" or "stable" part of the linker. In other embodiments, A can also be the "cleavable" part. In some

embodiments, A can be amide, carbamate, thioether via maleimide or bromoacetamide, triazole, oxime, or oxacarboline. The cleaved active agent or drug can retain the chemical structure of the active agent before cleavage, or can be modified as a result of cleavage. Moreover, depending on the desired therapeutic properties of the peptide-drug conjugate, such active agent can be active while linked to the peptide, remain active after cleavage or become inactivated, be inactive while linked to the peptide, or it can be activated upon cleavage. [0133] In some embodiments, peptide conjugates have stable linkers. A peptide of the disclosure can be expressed recombinantly or chemically synthesized. The peptide can be conjugated to a detectable agent or an active agent via a stable linker, such as an amide linkage or a carbamate linkage. The peptide can be conjugated to a detectable agent or an active agent via a stable linker, such as an amide bond using standard l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) or dicylcohexylcarbodiimide (DCC) based chemistry or thionyl chloride or phosphorous chloride- based bioconjugation chemistries. A stable linker may or may not be cleaved in buffer over extended periods of time (e.g., hours, days, or weeks). A stable linker may or may not be cleaved in body fluids such as plasma or synovial fluid over extended periods of time (e.g., hours, days, or weeks). A stable linker, may or may not be cleaved after exposure to enzymes, reactive oxygen species, other chemicals or enzymes that can be present in cells (e.g., macrophages), cellular compartments (e.g., endosomes and lysosomes), inflamed areas of the body (e.g., inflamed joints), tissues or body compartments. A stable linker may be cleaved by unknown mechanisms. A stable linker may or may not be cleaved in vivo but remains an active agent after peptide conjugation.

[0134] A peptide and drug conjugated via a linker can be described with the formula Peptide- A-

B-C-Drug, wherein the linker is A-B-C. A can be a stable amide link such as that formed by reacting an amine on the peptide with a linker containing a tetrafluorophenyl (TFP) ester or an

NHS ester. A can also be a stable carbamate linker such as that formed by reacting an amine on the peptide with an imidazole carbamate active intermediate formed by reaction of CDI with a hydroxyl on the linker. A can also be a stable secondary amine linkage such as that formed by reductive alkylation of the amine on the peptide with an aldehyde or ketone group on the linker.

A can also be a stable thioether linker formed using a maleimide or bromoacetamide in the linker with a thiol in the peptide, a triazole linker, a stable oxime linker, or a oxacarboline linker. B can be (-CH2-) _X - or a short PEG (-CH ₂ CH ₂ 0-) _X (x is 0-20) or other spacers or no spacer. C can be an amide bond formed with an amine or a carboxylic acid on the drug, a thioether formed between a maleimide on the linker and a sulfhydroyl on the drug, a secondary or tertiary amine, a carbamate, or other stable bonds. Any linker chemistry described in "Current ADC Linker

Chemistry," Jain et al. , Pharm Res, 2015 DOI 10. 1007/sl 1095-015-1657-7 can be used.

[0135] The resulting peptide conjugates can be administered to a human or animal

subcutaneously, intravenously, orally, or injected directly into a joint to treat disease. The peptide is not specifically cleaved from the detectable agent or active agent via a targeted mechanism.

The peptide can be degraded by mechanisms such as catabolism, releasing a drug that is modified or not modified form its native form (Antibody- Drug Conjugates: Design, Formulation, and

Physicochemical Stability, Singh, Luisi, and Pak. Pharm Res (2015) 32:3541-3571). The peptide drug conjugate exerts its pharmacological activity while still intact, or while partially or fully degraded, metabolized, or catabolized.

[0136] In some embodiments, peptide conjugates can have cleavable linkers. In some embodiments, a peptide and drug can be conjugated via a linker and can be described with the formula Peptide- A-B-C-Drug, wherein the linker is A-B-C. In some embodiments, A can be a stable amide link such as that formed by reacting an amine on the peptide with a linker containing a tetrafluorophenyl (TFP) ester or an NHS ester. In certain embodiments, A can also be a stable carbamate linker that is formed by an amine reaction on the peptide with an imidazole carbamate active intermediate formed by reaction of CDI with a hydroxyl on the linker. In other embodiments, A can also be a stable secondary amine linkage such as that formed by reductive alkylation of the amine on the peptide with an aldehyde or ketone group on the linker. In some embodiments, A can also be a stable thioether linker formed using a maleimide or

bromoacetamide in the linker with a thiol in the peptide, a triazole linker, a stable oxime linker, or an oxacarboline linker. B can be (-CH2-) _X - or a short PEG (-CH ₂ CH ₂ 0-) _X (x is 0-20) or other spacers or no spacer. C can be an ester bond to the hydroxyl or carboxylic acid on the drug, or a carbonate, hydrazone, or acylhydrazone, designed for hydrolytic cleavage. The hydrolytic rate of cleavage can be varied by varying the local environment around the bond, including carbon length (-CH2-)x, steric hindrance (including adjacent side groups such as methyl, ethyl, cyclic), hydrophilicity or hydrophobicity. In some embodiments, peptide conjugates can have a linear or cyclic ester linkage, which can include or do not include side chains such as methyl or ethyl groups. A linear ester linkage can be more susceptible to cleavage (such as by hydrolysis, an enzyme such as esterase, or other chemical reaction) than a cyclic ester due to steric hindrance or hydrophobicity/hydrophilicity effects. Likewise, side chains such as methyl or ethyl groups on the linear ester linkage can optionally make the linkage less susceptible to cleavage than without the side chains. In some embodiments, hydrolysis rate can be affected by local pH, such as lower pH in certain compartments of the body or of the cell such as endosomes and lysosomes or diseased tissues. In some embodiments, C can also be a pH sensitive group such as a hydrazone or oxime linkage. In other embodiments, C can be a disulfide bond designed to be released by reduction, such as by glutathione. In other embodiments, (or A-B-C) can be a peptidic linkage design for cleavabe by enzymes. Optionally, a self-immolating group such as pABC can be included to cause release of a free unmodified drug upon cleavage (Antibody-Drug Conjugates:

Design, Formulation, and Physicochemical Stability, Singh, Luisi, and Pak. Pharm Res (2015)

32:3541-3571). The linker can be cleaved by enzymes such as esterases, matrix

metalloproteinases, cathepsins such as cathepsin B, glucuronidases, a protease, or thrombin.

Alternatively, the bond designed for cleavage can be at A, rather than C, and C can be a stable bond or a cleavable bond. An alternative design can be to have stable linkers (such as amide or carbamate) at A and C and have a cleavable linker in B, such as a disulfide bond. The rate of reduction can be modulated by local effects such as steric hindrance from methyl or ethyl groups or modulating hydrophobicity/hydrophilicity. In some embodiments, peptide conjugates can have an ester carbonyl linkage, a long hydrocarbon linker, or carbamate linker, each of which can include hydrophilic groups, such as alcohols, acids, or ethers, or include a hydrocarbon side chain or other moiety that tunes the rate of cleavage. For example, the rate of hydrolysis can be faster with hydrophilic groups, such as alcohols, acids, or ethers, near an ester carbonyl. In another example, hydrophobic groups present as side chains or as a longer hydrocarbon linker can slow the cleavage rate of the ester. Likewise, cleavage of a carbamate group can also be tuned by hindrance, hydrophobicity, and the like. In another example, using a less labile linking group, such as a carbamate rather than an ester, can slow the cleavage rate of the linker.

[0137] Non- limiting examples of linkers include:

wherein each n is independently 0 to about 1,000; 1 to about 1,000; 0 to about 500; 1 to about 500; 0 to about 250; 1 to about 250; 0 to about 200; 1 to about 200; 0 to about 150; 1 to about 150; 0 to about 100; 1 to about 100; 0 to about 50; 1 to about 50; 0 to about 40; 1 to about 40; 0 to about 30; 1 to about 30; 0 to about 25; 1 to about 25; 0 to about 20; 1 to about 20; 0 to about 15; 1 to about 15; 0 to about 10; 1 to about 10; 0 to about 5; or 1 to about 5. In some embodiments, each n is independently 0, about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, or about 50. In some embodiments, m is 1 to about 1,000; 1 to about 500; 1 to about 250; 1 to about 200; 1 to about 150; 1 to about 100; 1 to about 50; 1 to about 40; 1 to about 30; 1 to about 25; 1 to about 20; 1 to about 15; 1 to about 10; or 1 to about 5. In some embodiments, m is 0, about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, or about 50.

[0138] In some cases a linker can be a succinic linker, and a drug can be attached to a peptide via an ester bond or an amide bond with two methylene carbons in between. In other cases, a linker can be any linker with both a hydroxyl group and a carboxylic acid, such as hydroxy hexanoic acid or lactic acid.

[0139] The linker can be a cleavable or a stable linker. The use of a cleavable linker permits release of the conjugated moiety (e.g., a therapeutic agent) from the peptide, e.g., after targeting to the cartilage. In some cases the linker is enzyme cleavable, e.g., a valine-citrulline linker. In some embodiments, the linker contains a self-immolating portion. In other embodiments, the linker includes one or more cleavage sites for a specific protease, such as a cleavage site for matrix metalloproteases (MMPs), thrombin, or cathepsin. Alternatively or in combination, the linker is cleavable by other mechanisms, such as via pH, reduction, or hydrolysis. A

hydro lytic ally labile linker, (amongst other cleavable linkers described herein) can be

advantageous in terms of releasing active agents from the peptide. For example, an active agent in a conjugate form with the peptide may not be active, but upon release from the conjugate after targeting to the cartilage, the active agent is active.

[0140] The rate of hydrolysis of the linker can be tuned. For example, the rate of hydrolysis of linkers with unhindered esters is faster compared to the hydrolysis of linkers with bulky groups next an ester carbonyl. A bulky group can be a methyl group, an ethyl group, a phenyl group, a ring, or an isopropyl group, or any group that provides steric bulk. In some cases, the steric bulk can be provided by the drug itself, such as by ketorolac when conjugated via its carboxylic acid. The rate of hydrolysis of the linker can be tuned according to the residency time of the conjugate in the cartilage. For example, when a peptide is cleared from the cartilage relatively quickly, the linker can be tuned to rapidly hydrolyze. In contrast, for example, when a peptide has a longer residence time in the cartilage, a slower hydrolysis rate can allow for extended delivery of an active agent. This can be important when the peptide is used to deliver a drug to the cartilage. "Programmed hydrolysis in designing paclitaxel prodrug for nanocarrier assembly" Sci Rep 2015, 5, 12023 Fu et al. , provides an example of modified hydrolysis rates.

Peptide Stability

[0141] A peptide of the present disclosure can be stable in various biological conditions, as well as during manufacturing, handling, storage, and other conditions in either a liquid or a dried state. Additionally, a peptide of the present disclosure can be resistant to enzymatic cleavage needed for peptide processing by the immune system. For example, any peptide of SEQ ID NO: 1 - SEQ ID NO: 564 can exhibit resistance to reducing agents, proteases, oxidative conditions, or acidic conditions.

[0142] In some cases, biologic molecules (such as peptides and proteins) can provide therapeutic functions, but such therapeutic functions are decreased or impeded by instability caused by the in vivo environment. (Moroz et al., Adv Drug Deliv Rev 101: 108-21 (2016), Mitragotri et al., Nat Rev Drug Discov 13(9):655-72 (2014), Bruno et al., Ther Deliv (11): 1443-67 (2013), Sinha et al., Crit Rev Ther Drug Carrier Syst. 24(l):63-92 (2007), Hamman et al., BioDrugs 19(3): 165-77 (2005)). Peptide degradation can be a result of a number of processes involving hydrolytic pathways, peptide oxidation such as oxidation of methionine (Met) residues, deamidation of asparagine (Asn) and glutamine (Gin) residues, and isomerization and hydrolysis of an adjacent asparagine (Asp) residue. (Manning et al., Pharmaceutical Research, Vol. 27 No. 4 (2010)). The amino acid immediately following the Asn or Gin residue can also affect the rate of deamidation, whereas: Asn-Gly, Asn-Ser, Asn-His, and Gln-Gly can be more likely to undergo deamidation. Additionally, the peptide bond adjacent to amino acids such as Asp can undergo hydrolysis with amino acid pairings such as Asp-Gly, Asp-Ser, Asp-Tyr, and Asp-Pro, which can be more likely to undergo hydrolysis. Oxidation of amino acid residues such as Met can form a sulfoxide species. The specific degradation reactions rates can vary for any given peptide or protein sequence.

[0143] Furthermore, the microenvironment within the molecular structure of the peptide, solvent accessibility, and conformational stability of each residue can impact the likelihood of peptide degradation. Therefore, by modifying a peptide sequence to reduce occurrence of such

degradation events, a the modified peptide or peptide-conjugate can have increased beneficial properties over unmodified peptides or peptide-drug conjugates, such as improved therapeutic efficacy, an increased safety profile, and can be less expensive to manufacture and develop. Key formulaic considerations that can prevent peptide decay can include the use of excipients, formulation at a desired pH, and storage under specific conditions (e.g., temperature, oxygen, light exposure, solid or liquid state, and container excipient materials). To circumvent degradation, peptide residues can be substituted with amino acids that increase stability, which can result in more efficacious and durable therapeutic peptides.

[0144] With respect to in vivo stability, the GI tract can contain a region of low pH (e.g., pH ~1), a reducing environment, or a protease-rich environment that can degrade peptides and proteins. Proteolytic activity in other areas of the body, such as the mouth, eye, lung, intranasal cavity, joint, skin, vaginal tract, mucous membranes, and serum, can also be an obstacle to the delivery of functionally active peptides and polypeptides. Additionally, the half-life of peptides in serum can be very short, in part due to proteases, such that the peptide can be degraded too quickly to have a lasting therapeutic effect when administering a therapeutic and safe dosing regimen.

Likewise, proteolytic activity in cellular compartments, such as lysosomes, and reduction activity in lysosomes and the cytosol can degrade peptides and proteins such that they may be unable to provide a therapeutic function on intracellular targets. Therefore, peptides that are resistant to reducing agents, proteases, and low pH may be able to provide enhanced therapeutic effects or enhance the therapeutic efficacy of co-formulated or conjugated active agents in vivo.

[0145] Additionally, oral delivery of drugs can be desirable in order to target certain areas of the body (e.g., disease in the GI tract such as colon cancer, irritable bowel disorder, infections, metabolic disorders, and constipation) despite the obstacles to the delivery of functionally active peptides and polypeptides presented by this method of administration. For example, oral delivery of drugs can increase compliance by providing a dosage form that is more convenient for patients to take as compared to parenteral delivery. Oral delivery can be useful in treatment regimens that have a large therapeutic window. Therefore, peptides that are resistant to reducing agents, proteases, and low pH can allow for oral delivery of peptides without nullifying their therapeutic function.

[0146] Peptide Resistance to Reducing Agents. Peptides of this disclosure can contain one or more cysteines, which can participate in disulfide bridges that can be integral to preserving the folded state of the peptide. Exposure of peptides to biological environments with reducing agents can result in unfolding of the peptide and loss of functionality and bioactivity. For example, glutathione (GSH) is a reducing agent that can be present in many areas of the body and in cells, and can reduce disulfide bonds. As another example, a peptide can become reduced upon cellular internalization during trafficking of a peptide across the gastrointestinal epithelium after oral administration a peptide can become reduced upon exposure to various parts of the GI tract. The GI tract can be a reducing environment, which can inhibit the ability of therapeutic molecules with disulfide bonds to have optimal therapeutic efficacy, due to reduction of the disulfide bonds. A peptide can also be reduced upon entry into a cell, such as after internalization by endo somes or lysosomes or into the cytosol, or other cellular compartments. Reduction of the disulfide bonds and unfolding of the peptide can lead to loss of functionality or affect key pharmacokinetic parameters such as bioavailability, peak plasma concentration, bioactivity, and half-life.

Reduction of the disulfide bonds can also lead to increased susceptibility of the peptide to subsequent degradation by proteases, resulting in rapid loss of intact peptide after administration. In some embodiments, a peptide that is resistant to reduction can remain intact and can impart a functional activity for a longer period of time in various compartments of the body and in cells, as compared to a peptide that is more readily reduced.

[0147] In certain embodiments, the peptides of this disclosure can be analyzed for the

characteristic of resistance to reducing agents to identify stable peptides. In some embodiments, the peptides of this disclosure can remain intact after being exposed to different molarities of reducing agents such as 0.00001M - 0.0001M, 0.0001M - 0.001M, 0.001M - 0.01M, 0.01 M - 0.05 M, 0.05 M - 0.1 M, for greater 15 minutes or more. In some embodiments, the reducing agent used to determine peptide stability can be dithiothreitol (DTT), Tris (2-carboxyethyl) phosphine HC1 (TCEP), 2-Mercaptoethanol, (reduced) glutathione (GSH), or any combination thereof. In some embodiments, at least 5%-10%, at least 10%-20%, at least 20%-30%, at least 30%-40%, at least 40%-50%, at least 50%-60%, at least 60%-70%, at least 70%-80%, at least 80%-90%, or at least 90%- 100% of the peptide remains intact after exposure to a reducing agent.

[0148] Peptide Resistance to Proteases. The stability of peptides of this disclosure can be determined by resistance to degradation by proteases. Proteases, also referred to as peptidases or proteinases, can be enzymes that can degrade peptides and proteins by breaking bonds between adjacent amino acids. Families of proteases with specificity for targeting specific amino acids can include serine proteases, cysteine proteases, threonine proteases, aspartic proteases, glutamic proteases, esterases, serum proteases, and asparagine proteases. Additionally, metalloproteases, matrix metalloproteases, elastase, carboxypeptidases, Cytochrome P450 enzymes, and cathepsins can also digest peptides and proteins. Proteases can be present at high concentration in blood, in mucous membranes, lungs, skin, the GI tract, the mouth, nose, eye, and in compartments of the cell. Misregulation of proteases can also be present in various diseases such as rheumatoid arthritis and other immune disorders. Degradation by proteases can reduce bioavailability, biodistribution, half-life, and bioactivity of therapeutic molecules such that they are unable to perform their therapeutic function. In some embodiments, peptides that are resistant to proteases can better provide therapeutic activity at reasonably tolerated concentrations in vivo.

[0149] In some embodiments, peptides of this disclosure can resist degradation by any class of protease. In certain embodiments, peptides of this disclosure resist degradation by pepsin (which can be found in the stomach), trypsin (which can be found in the duodenum), serum proteases, or any combination thereof. In certain embodiments, peptides of this disclosure can resist degradation by lung proteases (e.g., serine, cysteinyl, and aspartyl proteases, metalloproteases, neutrophil elastase, alpha- 1 antitrypsin, secretory leucoprotease inhibitor, elafin), or any combination thereof. In some embodiments, the proteases used to determine peptide stability can be pepsin, trypsin, chymotrypsin, or any combination thereof. In some embodiments, at least 5%- 10%, at least 10%-20%, at least 20%-30%, at least 30%-40%, at least 40%-50%, at least 50%- 60%, at least 60%-70%, at least 70%-80%, at least 80%-90%, or at least 90%- 100% of the peptide remains intact after exposure to a protease. Peptides of SEQ ID NO: 199, SEQ ID NO: 27, and SEQ ID NO: 108 can have particular structural qualities, which make them more resistant to protease degradation. For example, peptide of SEQ ID NO: 27 and SEQ ID NO: 109 exhibit the "hitchin" topology as described previously, which can be associated with resistance to protease and chemical degradation.

[0150] Peptide Stability in Acidic Conditions. Peptides of this disclosure can be administered in biological environments that are acidic. For example, after oral administration, peptides can experience acidic environmental conditions in the gastric fluids of the stomach and

gastrointestinal (GI) tract. The pH of the stomach can range from -1-4 and the pH of the GI tract ranges from acidic to normal physiological pH descending from the upper GI tract to the colon.

In addition, the vagina, late endosomes, and lysosomes can also have acidic pH values, such as less than pH 7. These acidic conditions can lead to denaturation of peptides and proteins into unfolded states. Unfolding of peptides and proteins can lead to increased susceptibility to subsequent digestion by other enzymes as well as loss of biological activity of the peptide.

[0151] In certain embodiments, the peptides of this disclosure can resist denaturation and degradation in acidic conditions and in buffers, which simulate acidic conditions. In certain embodiments, peptides of this disclosure can resist denaturation or degradation in buffer with a pH less than 1, a pH less than 2, a pH less than 3, a pH less than 4, a pH less than 5, a pH less than 6, a pH less than 7, or a pH less than 8. In some embodiments, peptides of this disclosure remain intact at a pH of 1-3. In certain embodiments, at least 5%-10%, at least 10%-20%, at least

20%-30%, at least 30%-40%, at least 40%-50%, at least 50%-60%, at least 60%-70%, at least

70%-80%, at least 80%-90%, or at least 90%- 100% of the peptide remains intact after exposure to a buffer with a pH less than 1, a pH less than 2, a pH less than 3, a pH less than 4, a pH less than 5, a pH less than 6, a pH less than 7, or a pH less than 8. In other embodiments, at least 5%- 10%, at least 10%-20%, at least 20%-30%, at least 30%-40%, at least 40%-50%, at least 50%- 60%, at least 60%-70%, at least 70%-80%, at least 80%-90%, or at least 90%- 100% of the peptide remains intact after exposure to a buffer with a pH of 1-3. In other embodiments, the peptides of this disclosure can be resistant to denaturation or degradation in simulated gastric fluid (pH 1-2). In some embodiments, at least 5-10%, at least 10%-20%, at least 20%-30%, at least 30%-40%, at least 40%-50%, at least 50%-60%, at least 60%-70%, at least 70%-80%, at least 80%-90%, or at least 90-100% of the peptide remains intact after exposure to simulated gastric fluid. In some embodiments, low pH solutions such as simulated gastric fluid or citrate buffers can be used to determine peptide stability.

[0152] Peptide Stability at High Temperatures. Peptides of this disclosure can be administered in biological environments with high temperatures. For example, after oral administration, peptides can experience high temperatures in the body. Body temperature can range from 36°C to 40°C. High temperatures can lead to denaturation of peptides and proteins into unfolded states. Unfolding of peptides and proteins can lead to increased susceptibility to subsequent digestion by other enzymes as well as loss of biological activity of the peptide. In some embodiments, a peptide of this disclosure can remain intact at temperatures from 25°C to 100°C. High temperatures can lead to faster degradation of peptides. Stability at a higher temperature can allow for storage of the peptide in tropical environments or areas where access to refrigeration is limited. In certain embodiments, 5%-100% of the peptide can remain intact after exposure to 25 °C for 6 months to 5 years. 5%-100% of a peptide can remain intact after exposure to 70°C for 15 minutes to 1 hour. 5%-100% of a peptide can remain intact after exposure to 100°C for 15 minutes to 1 hour. In other embodiments, at least 5%-10%, at least 10%-20%, at least 20%-30%, at least 30%-40%, at least 40%-50%, at least 50%-60%, at least 60%-70%, at least 70%-80%, at least 80%-90%, or at least 90%- 100% of the peptide remains intact after exposure to 25 °C for 6 months to 5 years. In other embodiments, at least 5%-10%, at least 10%-20%, at least 20%-30%, at least 30%-40%, at least 40%-50%, at least 50%-60%, at least 60%-70%, at least 70%-80%, at least 80%-90%, or at least 90%- 100% of the peptide remains intact after exposure to 70°C for 15 minutes to 1 hour. In other embodiments, at least 5%-10%, at least 10%-20%, at least 20%-30%, at least 30%-40%, at least 40%-50%, at least 50%-60%, at least 60%-70%, at least 70%-80%, at least 80%-90%, or at least 90%- 100% of the peptide remains intact after exposure to 100°C for 15 minutes to 1 hour.

[0153] In some embodiments, the peptide of the peptide active agent conjugate comprises a sequence that has at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least

97%, at least 99% or 100% sequence identity with any one of SEQ ID NO: 24 - SEQ ID NO: 274 or a fragment thereof. In some embodiments, the peptide of the peptide active agent conjugate comprises a sequence that has at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with any one of SEQ ID NO:

260 - SEQ ID NO: 274 or a fragment thereof. In some embodiments, the peptide of the peptide active agent conjugate comprises a sequence that has at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with any one of

SEQ ID NO: 314 - SEQ ID NO: 564 or a fragment thereof. In some embodiments, the peptide of the peptide active agent conjugate comprises a sequence that has at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with any one of SEQ ID NO: 550 - SEQ ID NO: 564 or a fragment thereof. In some embodiments, the peptide of the peptide active agent conjugate comprises a sequence of any one of SEQ ID NO: 24

- SEQ ID NO: 274 or a fragment thereof. In some embodiments, the peptide of the peptide active agent conjugate comprises a sequence of any one of SEQ ID NO: 260 - SEQ ID NO: 274 or a fragment thereof. In some embodiments, the peptide of the peptide active agent conjugate comprises a sequence of any one of SEQ ID NO: 314 - SEQ ID NO: 564 or a fragment thereof.

In some embodiments, the peptide of the peptide active agent conjugate comprises a sequence of any one of SEQ ID NO: 550 - SEQ ID NO: 564 or a fragment thereof. In some embodiments, the peptide comprises a sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with any one of

SEQ ID NO: 260- SEQ ID NO: 574. In some embodiments, the peptide comprises at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or

100% sequence identity with any one of SEQ ID NO: 550- SEQ ID NO: 564. In some embodiments, the peptide active agent conjugate or the peptide comprises a sequence of any one of SEQ ID NO: 1 - SEQ ID NO: 23 or a fragment thereof. In some embodiments, the peptide active agent conjugate or the peptide comprises a sequence of any one of SEQ ID NO: 275 -

SEQ ID NO: 297 or a fragment thereof. In some embodiments, the peptide active agent conjugate or the peptide comprises a sequence of any one of SEQ ID NO: 21 - SEQ ID NO: 23 or a fragment thereof. In some embodiments, the peptide active agent conjugate or the peptide comprises a sequence of any one of SEQ ID NO: 295 - SEQ ID NO: 297 or a fragment thereof.

In some embodiments, the peptide active agent conjugate or the peptide comprises a peptide with at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least

90%, at least 95%, at least, 97%, at least 98%, or at least 99% identical to any one of SEQ ID

NO: 494 - SEQ ID NO: 540 or at least 30%, at least 40%, at least 50%, at least 60%, at least

70%, at least 80%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NO: 204 - SEQ ID NO: 250. Pharmacokinetics of Peptides

[0154] The pharmacokinetics of any of the peptides of this disclosure can be determined after administration of the peptide via different routes of administration. For example, the

pharmacokinetic parameters of a peptide of this disclosure can be quantified after intravenous, subcutaneous, intramuscular, rectal, aerosol, parenteral, ophthalmic, pulmonary, transdermal, vaginal, optic, nasal, oral, sublingual, inhalation, dermal, intrathecal, intranasal, intra-articular, peritoneal, buccal, synovial, or topical administration. Peptides of the present disclosure can be analyzed by using tracking agents such as radiolabels or fluorophores. For example, a

radiolabeled peptide of this disclosure can be administered via various routes of administration. Peptide concentration or dose recovery in various biological samples such as plasma, urine, feces, any organ, skin, muscle, and other tissues can be determined using a range of methods including HPLC, fluorescence detection techniques (TECAN quantification, flow cytometry, iVIS), or liquid scintillation counting.

[0155] The methods and compositions described herein can relate to pharmacokinetics of peptide administration via any route to a subject. Pharmacokinetics can be described using methods and models, for example, compartmental models or noncompartmental methods. Compartmental models include but are not limited to monocompartmental model, the two compartmental model, the multicompartmental model or the like. Models can be divided into different compartments and can be described by the corresponding scheme. For example, one scheme is the absorption, distribution, metabolism and excretion (ADME) scheme. For another example, another scheme is the liberation, absorption, distribution, metabolism and excretion (LADME) scheme. In some aspects, metabolism and excretion can be grouped into one compartment referred to as the elimination compartment. For example, liberation can include liberation of the active portion of the composition from the delivery system, absorption includes absorption of the active portion of the composition by the subject, distribution includes distribution of the composition through the blood plasma and to different tissues, metabolism, which includes metabolism or inactivation of the composition and finally excretion, which includes excretion or elimination of the composition or the products of metabolism of the composition. Compositions administered intravenously to a subject can be subject to multiphasic pharmacokinetic profiles, which can include but are not limited to aspects of tissue distribution and metabolism/excretion. As such, the decrease in plasma or serum concentration of the composition is often biphasic, including, for example an alpha phase and a beta phase, occasionally a gamma, delta or other phase is observed

[0156] Pharmacokinetics includes determining at least one parameter associated with

administration of a peptide to a subject. In some aspects, parameters include at least the dose (D), dosing interval (τ), area under curve (AUC), maximum concentration (C _max ), minimum concentration reached before a subsequent dose is administered (Cmi _n ), minimum time (Tmin), maximum time to reach Cmax (T _max ), volume of distribution (V _d ), steady-state volume of distribution (V _ss ), back-extrapolated concentration at time 0 (Co), steady state concentration (C _ss ), elimination rate constant (k _e ), infusion rate (kj _n ), clearance (CL), bioavailability (f), fluctuation (%PTF) and elimination half-life (ti _/2 ).

[0157] In certain embodiments, the peptides of any of SEQ ID NO: 1 - SEQ ID NO 564 exhibit optimal pharmacokinetic parameters after oral administration. In other embodiments, the peptides of any of SEQ ID NO: 1 - SEQ ID NO: 564 exhibit optimal pharmacokinetic parameters after any route of administration, such as oral administration, inhalation, intranasal administration, topical administration, parenteral administration, intravenous administration, subcutaneous administration, intra-articular administration, intramuscular administration, intraperitoneal administration, transdermal administration, dermal administration, or any combination thereof.

[0158] In some embodiments any peptide of SEQ ID NO: 1 - SEQ ID NO: 564 exhibits an average T _max of 0.5 - 12 hours, or 1-48 hours at which the C _max is reached, an average

bioavailability in serum of 0.1%- 10% in the subject after administering the peptide to the subject by an oral route, an average bioavailability in serum of less than 0.1% after oral administration to a subject for delivery to the GI tract, an average bioavailability in serum of 10-100% after parenteral administration, an average t _½ of 0.1 hours - 168 hours, or 0.25 hours - 48 hours in a subject after administering the peptide to the subject, an average clearance (CL) of 0.5-100 L/hour or 0.5 - 50 L/hour of the peptide after administering the peptide to a subject, an average volume of distribution (V _d ) of 200 - 20,000 mL in the subject after systemically administering the peptide to the subject, or optionally no systemic uptake, any combination thereof.

Methods of Manufacture

[0159] Various expression vector/host systems can be utilized for the production of the recombinant expression of peptides described herein. Non- limiting examples of such systems include microorganisms such as bacteria transformed with recombinant bacteriophage DNA, plasmid DNA or cosmid DNA expression vectors containing a nucleic acid sequence encoding peptides or peptide fusion proteins/chimeric proteins described herein, yeast transformed with recombinant yeast expression vectors containing the aforementioned nucleic acid sequence, insect cell systems infected with recombinant virus expression vectors (e.g., baculo virus) containing the aforementioned nucleic acid sequence, plant cell systems infected with

recombinant virus expression vectors (e.g., cauliflower mosaic virus (CaMV), tobacco mosaic virus (TMV) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing the aforementioned nucleic acid sequence, or animal cell systems infected with recombinant virus expression vectors (e.g., adenovirus, vaccinia virus) including cell lines engineered to contain multiple copies of the aforementioned nucleic acid sequence, either stably amplified (e.g., CHO/dhfr, CHO/glutamine synthetase) or unstably amplified in double-minute chromosomes (e.g., murine cell lines). Disulfide bond formation and folding of the peptide could occur during expression or after expression or both.

[0160] A host cell can be adapted to express one or more peptides described herein. The host cells can be prokaryotic, eukaryotic, or insect cells. In some cases, host cells are capable of modulating the expression of the inserted sequences, or modifying and processing the gene or protein product in the specific fashion desired. For example, expression from certain promoters can be elevated in the presence of certain inducers (e.g., zinc and cadmium ions for

metallothionine promoters). In some cases, modifications (e.g., phosphorylation) and processing (e.g., cleavage) of peptide products can be important for the function of the peptide. Host cells can have characteristic and specific mechanisms for the post-translational processing and modification of a peptide. In some cases, the host cells used to express the peptides secretes minimal amounts of proteolytic enzymes.

[0161] In the case of cell- or viral-based samples, organisms can be treated prior to purification to preserve and/or release a target polypeptide. In some embodiments, the cells are fixed using a fixing agent. In some embodiments, the cells are lysed. The cellular material can be treated in a manner that does not disrupt a significant proportion of cells, but which removes proteins from the surface of the cellular material, and/or from the interstices between cells. For example, cellular material can be soaked in a liquid buffer or, in the case of plant material, can be subjected to a vacuum, in order to remove proteins located in the intercellular spaces and/or in the plant cell wall. If the cellular material is a microorganism, proteins can be extracted from the microorganism culture medium. Alternatively, the peptides can be packed in inclusion bodies. The inclusion bodies can further be separated from the cellular components in the medium. In some embodiments, the cells are not disrupted. A cellular or viral peptide that is presented by a cell or virus can be used for the attachment and/or purification of intact cells or viral particles. In addition to recombinant systems, Peptides can also be synthesized in a cell- free system using a variety of known techniques employed in protein and peptide synthesis.

[0162] In some cases, a host cell produces a peptide that has an attachment point for a drug. An attachment point could comprise a lysine residue, an N-terminus, a cysteine residue, a cysteine disulfide bond, or a non-natural amino acid. The peptide could also be produced synthetically, such as by solid-phase peptide synthesis, or solution-phase peptide synthesis. The peptide could be folded (formation of disulfide bonds) during synthesis or after synthesis or both. Peptide fragments could be produced synthetically or recombinantly and then joined together

synthetically, recombinantly, or via an enzyme.

[0163] FIG. 4 illustrates a schematic of a method of manufacturing a construct that expresses a peptide of the disclosure, such as the constructs illustrated in FIG. 3 and as described throughout the disclosure and in SEQ ID NO: 1 - SEQ ID NO: 564 provided herein.

[0164] In other aspects, the peptides of the present disclosure can be prepared by conventional solid phase chemical synthesis techniques, for example according to the Fmoc solid phase peptide synthesis method ("Fmoc solid phase peptide synthesis, a practical approach," edited by W. C. Chan and P. D. White, Oxford University Press, 2000), Boc solid phase peptide synthesis, or solution phase peptide synthesis. The disulfide bonds can be formed after cleavage from the resin, such as by air oxidation or a buffer system with a set pH range such as from 7-10 and can contain a redox system such as glutathione/oxidized glutathione or cysteine/cystine. The disulfide bonds can also be formed by selective protection and deprotection of specific cysteine residues followed by oxidation, or on the resin. The peptide can be purified, such as by reversed-phase chromatography at any one or more steps during the production process. The peptide can be isolated by lyophilization and can be in various salt forms, such as TFA salt or ammonium and acetate salt.

Pharmaceutical Compositions of Peptides

[0165] A pharmaceutical composition of the disclosure can be a combination of any peptide described herein with other chemical components, such as carriers, stabilizers, diluents, dispersing agents, suspending agents, thickening agents, antioxidants, solubilizers, buffers, osmolytes, salts, surfactants, amino acids, encapsulating agents, bulking agents, cryoprotectants, and/or excipients. The pharmaceutical composition facilitates administration of a peptide described herein to an organism. Pharmaceutical compositions can be administered in

therapeutically-effective amounts as pharmaceutical compositions by various forms and routes including, for example, intravenous, subcutaneous, intramuscular, rectal, aerosol, parenteral, ophthalmic, pulmonary, transdermal, vaginal, optic, nasal, oral, sublingual, inhalation, dermal, intrathecal, intranasal, intra-articular, and topical administration. A pharmaceutical composition can be administered in a local or systemic manner, for example, via injection of the peptide described herein directly into an organ, optionally in a depot.

[0166] Parenteral injections can be formulated for bolus injection or continuous infusion. The pharmaceutical compositions can be in a form suitable for parenteral injection as a sterile suspension, solution or emulsion in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Pharmaceutical formulations for parenteral administration include aqueous solutions of a peptide described herein in water soluble form. Suspensions of peptides described herein can be prepared as oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions can contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. The suspension can also contain suitable stabilizers or agents which increase the solubility and/or reduce the aggregation of such peptides described herein to allow for the preparation of highly concentrated solutions. Alternatively, the peptides described herein can be lyophilized or in powder form for re-constitution with a suitable vehicle, e.g., sterile pyrogen- free water, before use. In some embodiments, a purified peptide is administered intravenously.

[0167] A peptide of the disclosure can be applied directly to an organ, or an organ tissue or cells, such as brain or brain tissue or cancer cells, during a surgical procedure. The recombinant peptides described herein can be administered topically and can be formulated into a variety of topically administrable compositions, such as solutions, suspensions, lotions, gels, pastes, medicated sticks, balms, creams, and ointments. Such pharmaceutical compositions can contain solubilizers, stabilizers, tonicity enhancing agents, buffers and preservatives.

[0168] In practicing the methods of treatment or use provided herein, therapeutically-effective amounts of the peptide described herein described herein can be administered in pharmaceutical compositions to a subject suffering from a condition that affects the immune system. In some embodiments, the subject is a mammal such as a human. A therapeutically-effective amount can vary widely depending on the severity of the disease, the age and relative health of the subject, the potency of the compounds used, and other factors.

[0169] Pharmaceutical compositions can be formulated using one or more physiologically- acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the active compounds into preparations that can be used pharmaceutically. Formulation can be modified depending upon the route of administration chosen. Pharmaceutical compositions comprising a peptide described herein can be manufactured, for example, by expressing the peptide in a recombinant system, purifying the peptide, lyophilizing the peptide, mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or compression processes. The pharmaceutical compositions can include at least one pharmaceutically acceptable carrier, diluent, or excipient and compounds described herein as free-base or pharmaceutically- acceptable salt form.

[0170] Methods for the preparation of peptides described herein comprising the compounds described herein include formulating the peptide described herein with one or more inert, pharmaceutically-acceptable excipients or carriers to form a solid, semi-solid, or liquid composition. Solid compositions include, for example, powders, tablets, dispersible granules, capsules, cachets, and suppositories. These compositions can also contain minor amounts of nontoxic, auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, and other pharmaceutically-acceptable additives.

[0171] Non- limiting examples of pharmaceutically-acceptable excipients can be found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999), each of which is incorporated by reference in its entirety.

Administration of Pharmaceutical Compositions

[0172] A pharmaceutical composition of the disclosure can be a combination of any peptide described herein with other chemical components, such as carriers, stabilizers, diluents, dispersing agents, suspending agents, thickening agents, and/or excipients. The pharmaceutical composition facilitates administration of a peptide described herein to an organism.

Pharmaceutical compositions can be administered in therapeutically-effective amounts as pharmaceutical compositions by various forms and routes including, for example, intravenous, subcutaneous, intramuscular, rectal, aerosol, parenteral, ophthalmic, pulmonary, transdermal, vaginal, optic, nasal, oral, inhalation, dermal, intra-articular, intrathecal, intranasal, and topical administration. A pharmaceutical composition can be administered in a local or systemic manner, for example, via injection of the peptide described herein directly into an organ, optionally in a depot.

[0173] Parenteral injections can be formulated for bolus injection or continuous infusion. The pharmaceutical compositions can be in a form suitable for parenteral injection as a sterile suspension, solution or emulsion in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Pharmaceutical formulations for parenteral administration include aqueous solutions of a peptide described herein in

water-soluble form. Suspensions of peptides described herein can be prepared as oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions can contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. The suspension can also contain suitable stabilizers or agents which increase the solubility and/or reduce the aggregation of such peptides described herein to allow for the preparation of highly concentrated solutions. Alternatively, the peptides described herein can be lyophilized or in powder form for re-constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use. In some embodiments, a purified peptide is administered intravenously. A peptide described herein can be administered to a subject, home, target, migrates to, is retained by, and/or binds to, or be directed to an organ, e.g., the cartilage.

[0174] A peptide of the disclosure can be applied directly to an organ, or an organ tissue or cells, such as cartilage or cartilage tissue or cells, during a surgical procedure. The recombinant peptides described herein can be administered topically and can be formulated into a variety of topically administrable compositions, such as solutions, suspensions, lotions, gels, pastes, medicated sticks, balms, creams, and ointments. Such pharmaceutical compositions can contain solubilizers, stabilizers, tonicity enhancing agents, buffers and preservatives.

[0175] In practicing the methods of treatment or use provided herein, therapeutically-effective amounts of the peptide described herein described herein are administered in pharmaceutical compositions to a subject suffering from a condition. In some instances the pharmaceutical composition will affect the physiology of the animal, such as the immune system, inflammatory response, or other physiologic affect. In some embodiments, the subject is a mammal such as a human. A therapeutically-effective amount can vary widely depending on the severity of the disease, the age and relative health of the subject, the potency of the compounds used, and other factors.

[0176] Pharmaceutical compositions can be formulated using one or more physiologically- acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the active compounds into preparations that can be used pharmaceutically. Formulation can be modified depending upon the route of administration chosen. Pharmaceutical compositions comprising a peptide described herein can be manufactured, for example, by expressing the peptide in a recombinant system, purifying the peptide, lyophilizing the peptide, mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or compression processes. The pharmaceutical compositions can include at least one pharmaceutically acceptable carrier, diluent, or excipient and compounds described herein as free-base or pharmaceutically- acceptable salt form.

[0177] Methods for the preparation of peptides described herein comprising the compounds described herein include formulating the peptide described herein with one or more inert, pharmaceutically-acceptable excipients or carriers to form a solid, semi-solid, or liquid composition. Solid compositions include, for example, powders, tablets, dispersible granules, capsules, cachets, and suppositories. These compositions can also contain minor amounts of nontoxic, auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, and other pharmaceutically-acceptable additives.

[0178] Non- limiting examples of pharmaceutically-acceptable excipients can be found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E. , Remington 's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999), each of which is incorporated by reference in its entirety.

Use of Peptide in Imaging and Surgical Methods

[0179] The present disclosure generally relates to peptides that home, target, migrate to, are retained by, accumulate in, and/or bind to, or are directed to specific regions, tissues, structures, or cells within the body and methods of using such peptides. These peptides have the ability to contact the cartilage, which makes them useful for a variety of applications. In particular, the peptides can have applications in site-specific modulation of bio molecules to which the peptides are directed to. End uses of such peptides can include, for example, imaging, research, therapeutics, theranostics, pharmaceuticals, chemotherapy, chelation therapy, targeted drug delivery, and radiotherapy. Some uses can include targeted drug delivery and imaging.

[0180] In some embodiments, the present disclosure provides a method for detecting a cancer, cancerous tissue, or tumor tissue, the method comprising the steps of contacting a tissue of interest with a peptide of the present disclosure, wherein the peptide is conjugated to a detectable agent and measuring the level of binding of the peptide, wherein an elevated level of binding, relative to normal tissue, is indicative that the tissue is a cancer, cancerous tissue or tumor tissue.

[0181] In some embodiments, the disclosure provides a method of imaging an organ or body region or region, tissue or structure of a subject, the method comprising administrating to the subject the peptide or a pharmaceutical composition disclosed herein and imaging the subject. In some embodiments such imaging is used to detect a condition associated with cartilage, or a function of the cartilage. In some cases the condition is an inflammation, a cancer, a degradation, a growth disturbance, genetic, a tear or an injury, or another suitable condition. In some cases the condition is a chondrodystrophy, a traumatic rupture or detachment, pain following surgery in regions of the body containing cartilage, costochondritis, herniation, polychondritis, arthritis, osteoarthritis, rheumatoid arthritis, ankylosing spondylitis (AS), Systemic Lupus Erythematosus (SLE or "Lupus"), Psoriatic Arthritis (PsA), gout, achondroplasia, or another suitable condition. In some case the condition is associated with a cancer or tumor of the cartilage. In some cases the condition is a type of chondroma or chondrosarcoma, whether metastatic or not, or another suitable condition. In some embodiments, such as those associated with cancers, the imaging may be associated with surgical removal of the diseased region, tissue, structure or cell of a subject.

[0182] Furthermore, the present disclosure provides methods for intraoperative imaging and resection of a diseased or inflamed tissue, cancer, cancerous tissue, or tumor tissue using a peptide of the present disclosure conjugated with a detectable agent. In some embodiments, the diseased or inflamed tissue, cancer, cancerous tissue, or tumor tissue is detectable by

fluorescence imaging that allows for intraoperative visualization of the cancer, cancerous tissue, or tumor tissue using a peptide of the present disclosure. In some embodiments, the peptide of the present disclosure is conjugated to one or more detectable agents. In a further embodiment, the detectable agent comprises a fluorescent moiety coupled to the peptide. In another embodiment, the detectable agent comprises a radionuclide. In some embodiments, imaging is achieved during open surgery. In further embodiments, imaging is accomplished using endoscopy or other noninvasive surgical techniques.

Treatment of Cartilage Disorders

[0183] The term "effective amount," as used herein, can refer to a sufficient amount of an agent or a compound being administered which will relieve to some extent one or more of the symptoms of the disease or condition being treated. The result can be reduction and/or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. Compositions containing such agents or compounds can be administered for

prophylactic, enhancing, and/or therapeutic treatments. An appropriate "effective" amount in any individual case can be determined using techniques, such as a dose escalation study.

[0184] The methods, compositions, and kits of this disclosure can comprise a method to prevent, treat, arrest, reverse, or ameliorate the symptoms of a condition. The treatment can comprise treating a subject (e.g., an individual, a domestic animal, a wild animal or a lab animal afflicted with a disease or condition) with a peptide of the disclosure. In treating a disease, the peptide can contact the cartilage of a subject. The subject can be a human. A subject can be a human; a non- human primate such as a chimpanzee, or other ape or monkey species; a farm animal such as a cattle, horse, sheep, goat, swine; a domestic animal such as a rabbit, dog, and cat; a laboratory animal including a rodent, such as a rat, mouse and guinea pig, or the like. A subject can be of any age. A subject can be, for example, an elderly adult, adult, adolescent, pre-adolescent, child, toddler, infant, or fetus in utero.

[0185] Treatment can be provided to the subject before clinical onset of disease. Treatment can be provided to the subject after clinical onset of disease. Treatment can be provided to the subject after 1 day, 1 week, 6 months, 12 months, or 2 years or more after clinical onset of the disease. Treatment may be provided to the subject for more than 1 day, 1 week, 1 month, 6 months, 12 months, 2 years or more after clinical onset of disease. Treatment may be provided to the subject for less than 1 day, 1 week, 1 month, 6 months, 12 months, or 2 years after clinical onset of the disease. Treatment can also include treating a human in a clinical trial. A treatment can comprise administering to a subject a pharmaceutical composition, such as one or more of the

pharmaceutical compositions described throughout the disclosure. A treatment can comprise a once daily dosing. A treatment can comprise delivering a peptide of the disclosure to a subject, either parenterally, intravenously, subcutaneously, intramuscularly, by inhalation, dermally, intra-articular injection, orally, intrathecally, transdermally, intranasally, via a peritoneal route, or directly onto or into a joint, e.g., via topical, intra-articular injection route or injection route of application. A treatment can comprise administering a peptide- active agent complex to a subject, either parenterally, intravenously, subcutaneously, intramuscularly, by inhalation, dermally, intra-articular injection, orally, intrathecally, transdermally, intranasally, via a peritoneal route, or directly onto or into a joint or directly onto, near or into the cartilage, e.g., via topical, intraarticular injection route or injection route of application.

[0186] Types of cartilage diseases or conditions that can be treated with a peptide of the disclosure can include inflammation, pain management, anti-infective, pain relief, anti-cytokine, cancer, injury, degradation, genetic basis, remodeling, hyperplasia, surgical injury/trauma, or the like. Diseases or conditions of bone adjacent to cartilage can also be treated with a peptide of the disclosure. Examples of cartilage diseases or conditions that can be treated with a peptide of the disclosure include Costochondritis, Spinal disc herniation, Relapsing polychondritis, Injury to the articular cartilage, any manner of rheumatic disease (e.g., Rheumatoid Arthritis (RA), ankylosing spondylitis (AS), Systemic Lupus Erythematosus (SLE or "Lupus"), Psoriatic Arthritis (PsA),

Osteoarthritis, Gout, and the like), Herniation, Achondroplasia, Benign or non-cancerous chondroma, Malignant or cancerous chondrosarcoma, Chondriodystrophies, Chondromalacia patella, Costochondritis, Halus rigidus, Hip labral tear, Osteochondritis dssecans,

Osteochondrodysplasias, Torn meniscus, Pectus carinatum, Pectus excavatum, Chondropathy,

Chondromalacia, Polychondritis, Relapsing Polychondritis, Slipped epiphysis, Osteochondritis

Dissecans, Chondrodysplasia, Costochondritis, Perichondritis, Osteochondroma, Knee osteoarthritis, Finger osteoarthritis, Wrist osteoarthritis, Hip osteoarthritis, Spine osteoarthritis,

Chondromalacia, Osteoarthritis Susceptibility, Ankle Osteoarthritis, Spondylosis, Secondary chondrosarcoma, Small and unstable nodules as seen in osteoarthritis, Osteochondroses, Primary chondrosarcoma, Cartilage disorders, scleroderma, collagen disorders, Chondrodysplasia, Tietze syndrome, Dermochondrocorneal dystrophy of Francois, Epiphyseal dysplasia multiple 1, Epiphyseal dysplasia multiple 2, Epiphyseal dysplasia multiple 3, Epiphyseal dysplasia multiple 4, Epiphyseal dysplasia multiple 5, Ossified Ear cartilages with Mental deficiency, Muscle Wasting and Bony Changes, Periosteal chondrosarcoma, Carpotarsal osteochondromatosis, Achondroplasia, Genochondromatosis II, Genochondromatosis, Chondrodysplasia— disorder of sex development, Chondroma, Chordoma, Atelosteogenesis, type 1, Atelosteogenesis Type III, Atelosteogenesis, type 2, Pyknoachondrogenesis, Osteoarthropathy of fingers familial,

Dyschondrosteosis - nephritis, Coloboma of Alar-nasal cartilages with telecanthus, Alar cartilages hypoplasia - coloboma - telecanthus, Pierre Robin syndrome - fetal chondrodysplasia, Dysspondyloenchondromatosis, Achondroplasia regional - dysplasia abdominal muscle, Osteochondritis Dissecans, Familial Articular Chondrocalcinosis, Tracheobronchomalacia, Chondritis, Dyschondrosteosis, Jequier-Kozlowski-skeletal dysplasia, Chondrodystrophy, Cranio osteoarthropathy, Tietze's syndrome, Hip dysplasia - ecchondromata, Bessel-Hagen disease, Chondromatosis (benign), Enchondromatosis (benign), Chondrocalcinosis due to apatite crystal deposition, Meyenburg-Altherr-Uehlinger syndrome, Enchondromatosis-dwarfism-deafness, premature growth plate closure (e.g., due to dwarfism, injury, therapy such as retinoid therapy for adolescent acne, or ACL repair), Astley- Kendall syndrome, Synovial osteochondromatosis, Severe achondroplasia with developmental delay and acanthosis nigricans, Chondrocalcinosis, Stanescu syndrome, Familial osteochondritis dissecans, Achondrogenesis type 1A,

Achondrogenesis type 2, Achondrogenesis, Langer- Saldino Type, Achondrogenesis type IB, Achondrogenesis type 1A and IB, Type II Achondrogenesis-Hypochondrogenesis,

Achondrogenesis, Achondrogenesis type 3, Achondrogenesis type 4, Chondrocalcinosis 1, Chondrocalcinosis 2, Chondrocalcinosis familial articular, Diastrophic dysplasia,

Fibrochondrogenesis, Hypochondroplasia, Keutel syndrome, Maffucci Syndrome, Osteoarthritis Susceptibility 6, Osteoarthritis Susceptibility 5, Osteoarthritis Susceptibility 4, Osteoarthritis Susceptibility 3, Osteoarthritis Susceptibility 2, Osteoarthritis Susceptibility 1,

Pseudoachondroplasia, Cauliflower ear, Costochondritis, Growth plate fractures, Pectus excavatum, septic arthritis, gout, pseudogout (calcium pyrophosphate deposition disease or CPPD), gouty arthritis, bacterial, viral, or fungal infections in or near the joint, bursitis, tendinitis, arthropathies, or a joint disease condition. Examples of bone diseases or conditions that can be treated with a peptide of the disclosure include osteopenia, post-menopausal bone loss, bone maintenance, bone fracture, arthroplasty recovery, osteoporosis, bone loss due to metastatic cancer, fractures due to bone loss (e.g., hip fractures in patients with osteoporosis), pathological fracture, or atypical fracture.

[0187] In some embodiments, a peptide or peptide conjugate of this disclosure can be

administered to a subject in order to target, an arthritic joint. In other embodiments, a peptide or peptide conjugate of this disclosure can be administered to a subject in order to treat an arthritic joint.

[0188] In some embodiments, the present disclosure provides a method for treating a cancer, the method comprising administering to a subject in need thereof an effective amount of a peptide of the present disclosure.

[0189] In some embodiments, the present disclosure provides a method for treating a cancer, the method comprising administering to a patient in need thereof an effective amount of a

pharmaceutical composition comprising a peptide of the present disclosure and a

pharmaceutically acceptable carrier.

[0190] In some embodiments, the peptides of the present disclosure can be used to treat chondrosarcoma. Chondrosarcoma is a cancer of cartilage producing cells and is often found in bones and joints. It falls within the family of bone and soft-tissue sarcomas. In certain

embodiments, administration of a peptide or peptide conjugate of the present disclosure can be used to image and diagnose or target and treat a subject with chondrosarcoma. The

administration of a peptide or peptide conjugate of the present disclosure can be used in combination with ablative radiotherapy or proton therapy to treat chondrosarcoma. The subject can be a human or an animal.

[0191] In some embodiments, a peptide or peptide conjugate of this disclosure can be used to treat Chordoma. In certain embodiments, administration of a peptide or peptide conjugate of the present disclosure can be used to image and diagnose or target and treat a subject with chordoma. The administration of a peptide or peptide conjugate of the present disclosure can be used in combination with a tyrosine kinase inhibitor, such as imatinib mesylate, and ablative

radiotherapy or proton therapy to treat chordoma. The administration of a peptide or peptide conjugate of the present disclosure can be used in combination with an antivascular agent such as bevacizumab and an epidermal growth factor receptor inhibitor such as erlotinib to treat chordoma. The subject can be a human or an animal.

[0192] In some embodiments, the present disclosure provides a method for inhibiting invasive activity of cells, the method comprising administering an effective amount of a peptide of the present disclosure to a subject.

[0193] In some embodiments, the peptides of the present disclosure are conjugated to one or more therapeutic agents. In further embodiments, the therapeutic agent is a chemotherapeutic, anti-cancer drug, or anti-cancer agent selected from, but are not limited to: anti- inflammatories, such as for example a glucocorticoid, a corticosteroid, a protease inhibitor, such as for example collagenase inhibitor or a matrix metalloprotease inhibitor (i.e., MMP-13 inhibitor), an amino sugar, vitamin (e.g., Vitamin D), and antibiotics, antiviral, or antifungal, a statin, an immune modulator, radioisotopes, toxins, enzymes, sensitizing drugs, nucleic acids, including interfering RNAs, antibodies, anti-angiogenic agents, cisplatin, anti-metabolites, mitotic inhibitors, growth factor inhibitors, paclitaxel, temozolomide, topotecan, fluorouracil, vincristine, vinblastine, procarbazine, decarbazine, altretamine, methotrexate, mercaptopurine, thioguanine, fludarabine phosphate, cladribine, pento statin, cytarabine, azacitidine, etoposide, teniposide, irinotecan, docetaxel, doxorubicin, daunorubicin, dactinomycin, idarubicin, plicamycin, mitomycin, bleomycin, tamoxifen, flutamide, leuprolide, goserelin, aminogluthimide, anastrozole, amsacrine, asparaginase, mitoxantrone, mitotane and amifostine, and their equivalents, as well as photo- ablation. Some of these active agents induce programmed cell death such as apoptosis in target cells and thereby improve symptoms or ameliorate disease. Apoptosis can be induced by many active agents, including, for example, chemotherapeutics, anti- inflammatories, corticosteroids, NSAIDS, tumor necrosis factor alpha (TNF-a) modulators, tumor necrosis factor receptor (TNFR) family modulators. In some embodiments, peptides of this disclosure can be used to target active agents to pathways of cell death or cell killing, such as caspases, apoptosis activators and inhibitors, XBP-1, Bcl-2, Bcl-Xl, Bcl-w, and other disclosed herein. In other embodiments, the therapeutic agent is any nonsteroidal anti- inflammatory drug (NSAID). The NSAID can be any heterocyclic acetic acid derivatives such as ketorolac, indomethacin, etodolac, or tolemetin, any propionic acid derivatives such as naproxen, any enolic acid derivatives, any anthranilic acid derivatives, any selective COX-2 inhibitors such as celecoxib, any sulfonanilides, any salicylates, aceclofenac, nabumetone, sulindac, diclofenac, or ibuprofen. In other

embodiments, the therapeutic agent is any steroid, such as dexamethasone, budesonide, beclomethasone monopropionate, desciclesonide, triamcinolone, cortisone, prednisone, prednisolone, triamcinolone hexacetonide, or methylprednisolone. In other embodiments, the therapeutic agent is a pain reliever, such as acetaminophen, opioids, local anesthetics, antidepressants, glutamate receptor antagonists, adenosine, or neuropeptides. In some embodiments, a treatment consists of administering a combination of any of the above therapeutic agents and a peptide conjugate, such as a treatment in which both a dexamethasone-peptide conjugate and an NSAID are administered to a patient. Peptides of the current disclosure that target the cartilage can be used to treat the diseases conditions as described herein, for example, any diseases or conditions including tears, injuries (i.e., sports injuries), genetic factors, degradation, thinning, inflammation, cancer or any other disease or condition of the cartilage or to target

therapeutically-active substances to treat these diseases amongst others. In other cases, a peptide of the disclosure can be used to treat traumatic rupture, detachment, chostochondritis, spinal disc herniation, relapsing and non-relapsing polychondritis, injury to the articular cartilage, osteoarthritis, arthritis or achondroplasia. In some cases, the peptide or peptide- active agent can be used to target cancer in the cartilage, for example benign chondroma or malignant chondrosarcoma, by contacting the cartilage by diffusion into chondrocytes and then having antitumor function, targeted toxicity, inhibiting metastases, etc. As well, such peptide or peptide- active agent can be used to label, detect, or image such cartilage lesions, including tumors and metastases amongst other lesions, which may be removed through various surgical techniques or by targeting with peptide- active agents that induce programmed cell death or kill cells.

[0194] Venom or toxin derived peptide(s), peptides, modified peptides, labeled peptides, peptide- active agent conjugates and pharmaceutical compositions described herein can be administered for prophylactic and/or therapeutic treatments. In therapeutic applications, the composition can be administered to a subject already suffering from a disease or condition, in an amount sufficient to cure or at least partially arrest the symptoms of the disease or condition, or to cure, heal, improve, or ameliorate the condition. Such peptides described herein can also be administered to prevent (either in whole or in part), lessen a likelihood of developing, contracting, or worsening a condition. Amounts effective for this use can vary based on the severity and course of the disease or condition, previous therapy, the subject's health status, weight, response to the drugs, and the judgment of the treating physician. Venom or toxin derived peptide(s), peptides, modified peptides, labeled peptides, peptide- active agent conjugates and pharmaceutical compositions described herein can allow for targeted homing of the peptide and local delivery of any conjugate. For example, a peptide conjugated to a steroid allows for local delivery of the steroid, which is significantly more effective and less toxic than traditional systemic steroids. A peptide conjugated to an NSAID is another example. In this case, the peptide conjugated to an NSAID allows for local delivery of the NSAID, which allows for administration of a lower NSAID dose and is subsequently less toxic. By delivering an active agent to the joint, pain relief can be more rapid, may be more long lasting, and can be obtained with a lower systemic dose and off-site undesired effects than with systemic dosing without targeting.

[0195] Peptides of the current disclosure that target the cartilage can be used to treat or manage pain associated with a cartilage injury or disorder, or any other cartilage or joint condition as described herein. The peptides can be used either directly or as carriers of active drugs, peptides, or molecules. For example, since ion channels can be associated with pain and can be activated in disease states such as arthritis, peptides that interact with ion channels can be used directly to reduce pain. In another embodiment, the peptide is conjugated to an active agent with anti- inflammatory activity, in which the peptide acts as a carrier for the local delivery of the active agent to reduce pain.

[0196] In some embodiments, the peptides described herein provide a method of treating a cartilage condition of a subject, the method comprising administering to the subject a therapeutically-effective amount of a peptide comprising the sequence SEQ ID NO: 1 or fragment thereof. In some embodiments, the peptides described herein provide a method of treating a cartilage condition of a subject, the method comprising administering to the subject a peptide of any one of SEQ ID NO: 2 -SEQ ID NO: 564 or fragment thereof.

Treatment of Kidney Disorders

[0197] In some embodiments, peptides of this disclosure that home, target, are directed to, migrate to, are retained by, accumulate in, or bind to specific regions, tissues, structures or cells of the kidneys can be used to treat a kidney disorder. In other embodiments, peptides are used in peptide conjugates of the present disclosure to deliver an active agent for treatment of a kidney disorder.

[0198] In some embodiments, the peptides and peptide-conjugates of the present disclosure are used to treat a condition of the kidney, or a region, tissue, structure, or cell thereof. In certain embodiments, the condition is associated with kidney, or a function of a subject's kidneys. The present disclosure encompasses various acute and chronic renal diseases, including glomerular, tubule-interstitial, and microvascular diseases. Examples of conditions applicable to the present disclosure include but are not limited to: hypertensive kidney damage, acute kidney diseases and disorders (AKD), acute kidney injury (AKI) due to ischemia-reperfusion injury, drug treatment such as chemotherapy, cardiovascular surgery, surgery, medical interventions or treatment, radiocontrast nephropathy, or induced by cisplatin or carboplatin, which can be treated prophylactically, established AKI including ischemic renal injury, endotoxemia-induced AKI, endotoxemia sepsis syndrome, or established nephrotoxic AKI (e.g., rhabdomyolysis, radiocontrast nephropathy, cisplatin/carboplatin AKI, aminoglycoside nephrotoxicity), end stage renal disease, acute and rapidly progressive glomerulonephritis, acute presentations of nephrotic syndrome, acute pyelonephritis, acute renal failure, chronic glomerulonephritis, chronic heart failure, chronic interstitial nephritis, graft versus host disease after renal transplant, chronic kidney disease (CKD) such as diabetic nephropathy, hypertensive nephrosclerosis, idiopathic chronic glomerulonephritis (e.g., focal glomerular sclerosis, membranous nephropathy, membranoprohferative glomerulonephritis, minimal change disease transition to chronic disease, anti-GBM disease, rapidly progressive cresentic glomerulonephritis, IgA nephropathy), secondary chronic glomerulonephritis (e.g., systemic lupus, polyarteritis nodosa, scleroderma, amyloidosis, endocarditis), hereditary nephropathy (e.g., polycystic kidney disease, Alport's syndrome), interstitial nephritis induced by drugs (e.g., Chinese herbs, NSAIDs), multiple myeloma or sarcoid, or renal transplantation such as donor kidney prophylaxis (treatment of donor kidney prior to transplantation), treatment post transplantation to treat delayed graft function, acute rejection, or chronic rejection, chronic liver disease, chronic pyelonephritis, diabetes, diabetic kidney disease, fibrosis, focal segmental glomerulosclerosis, Goodpasture's disease, hypertensive nephrosclerosis, IgG4-related renal disease, interstitial inflammation, lupus nephritis, nephritic syndrome, partial obstruction of the urinary tract, polycystic kidney disease, progressive renal disease, renal cell carcinoma, renal fibrosis, and vasculitis. For example, in certain embodiments, the peptides and peptide-conjugates of the present disclosure are used to reduce acute kidney injury in order to prevent it from progressing to chronic kidney disease.

[0199] Alternatively or in combination, in some embodiments, the peptide and peptide- conjugates of the present disclosure are used to elicit a protective response such as ischemic preconditioning and/or acquired cytoresistance in a kidney of the subject. In some embodiments, ischemic preconditioning and/or acquired cytoresistance is induced by administering an agent (e.g., a peptide or peptide-conjugate of the present disclosure) that upregulates the expression of protective stress proteins, such as antioxidants, anti- inflammatory proteins, or protease inhibitors. In certain embodiments, the induced response protects the kidney by preserving kidney function in whole or in part and/or by reducing injury to renal tissues and cells, e.g., relative to the situation where no protective response is induced. The peptides and peptide-conjugates of the present disclosure can provide certain benefits compared to other agents for inducing ischemic preconditioning and/or acquired cytoresistance, such as a well-defined chemical structure and avoidance of low pH precipitation.

[0200] In some embodiments, the protective response is induced in order to protect the kidney or tissues or cells thereof from an injury or insult that is predicted to occur (e.g., associated with a planned event such as a medical procedure, is likely to occur due to a condition in the subject) or has already occurred. In certain embodiments, the induced response prevents or reduces the extent of damage to the kidney or tissues or cells thereof caused by the injury or insult. For instance, in certain embodiments, the peptides and peptide-conjugates induce acquired cytoresistance by activating protective pathways and/or upregulating expression of protective stress proteins. Optionally, the peptides and peptide-conjugates are capable of inducing such protective responses while causing minimal or no injury to the kidney.

[0201] In various embodiments, the injury or insult is associated with one or more of: surgery, radiocontrast imaging, cardiopulmonary bypass, balloon angioplasty, induced cardiac or cerebral ischemic-reperfusion injury, organ transplantation, sepsis, shock, low blood pressure, high blood pressure, kidney hypoperfusion, chemotherapy, drug administration, nephrotoxic drug

administration, blunt force trauma, puncture, poison, or smoking. For instance, in certain embodiments, the injury or insult is associated with a medical procedure that has been or will be performed on the subject, such as one or more of: surgery, radiocontrast imaging, cardiopulmonary bypass, balloon angioplasty, induced cardiac or cerebral ischemic-reperfusion injury, organ transplantation, chemotherapy, drug administration, or nephrotoxic drug

administration.

[0202] In some embodiments, the peptide itself exhibits a renal therapeutic effect. For example, in certain embodiments, the cystine-dense peptide interacts with a renal ion channel, inhibits a protease, has antimicrobial activity, has anticancer activity, has anti- inflammatory activity, induces ischemic preconditioning or acquired cytoresistance, or produces a protective or therapeutic effect on a kidney of the subject, or a combination thereof. Optionally, the renal therapeutic effect exhibited by the peptide is a renal protective effect or renal prophylactic effect (e.g., ischemic preconditioning or acquired cytoresistance) that protects the kidney or a tissue or cell thereof from an upcoming injury or insult.

[0203] For example, in certain embodiments, a peptide of the present disclosure activates protective pathways and/or upregulates expression of protective stress proteins in the kidney or tissues or cells thereof. As another example, in certain embodiments, a peptide of the present disclosure accesses and suppresses intracellular injury pathways. In yet another example, in certain embodiments, a peptide of the present disclosure inhibits interstitial inflammation and prevents renal fibrosis. As a further example, in certain embodiments, a peptide of the present disclosure is administered prior to or currently with the administration of a nephrotoxic agent (e.g., aminoglycoside antibiotics such as gentamicin and minocycline, chemotherapeutics such as cisplatin, immunoglobulins or fragments thereof, mannitol, NSAIDs such as ketorolac or ibuprofen, cyclosporin, cyclophosphamide, radiocontrast dyes) in order to minimize its damaging effects, e.g., by blocking megalin-cubulin binding sites so that the nephrotoxic agent passes through the kidneys.

[0204] In some embodiments, the present disclosure provides that any peptide of the disclosure including SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564 can as a peptide conjugate with an active agent for treatment of a kidney disorder. For example, a peptide of SEQ ID NO: 27, SEQ ID NO: 108, or SEQ ID NO: 199 can be conjugated to an active agent and administered to a subject in need thereof to treat a kidney disorder.

[0205] In some embodiments, homing of a peptide of this disclosure to cartilage or the kidneys can be assessed in an animal model such as those described in Alves et al. (Clin Rev Allergy Immunol. 2016 Aug;51(l):27-47. doi: 10. 1007/sl2016-015-8522-7), Kuyinu et al. (J Orthop Surg Res. 2016 Feb 2;11: 19. doi: 10. 1186/sl3018-016-0346-5), Li et al. (Exp Biol Med

(Maywood). 2015 Aug;240(8): 1029-38. doi: 10. 1177/1535370215594583), and Mullins et al. (Dis Model Mech. 2016 Dec 1;9(12): 1419-1433), all of which are incorporated herein by reference. [0206] Multiple peptides described herein can be administered in any order or simultaneously. In some cases, multiple functional fragments of peptides derived from toxins or venom can be administered in any order or simultaneously. If simultaneously, the multiple peptides described herein can be provided in a single, unified form, such as an intravenous injection, or in multiple forms, such as subsequent intravenous dosages.

[0207] Peptides can be packaged as a kit. In some embodiments, a kit includes written instructions on the use or administration of the peptides.

EXAMPLES

[0208] The following examples are included to further describe some embodiments of the present disclosure, and should not be used to limit the scope of the disclosure.

EXAMPLE 1

Manufacture of Peptides

[0209] The peptide sequence was reverse-translated into DNA, synthesized, and cloned in-frame with siderocalin using standard molecular biology techniques. (M. R. Green, Joseph Sambrook. Molecular Cloning. 2012 Cold Spring Harbor Press.). The resulting construct was packaged into a lentivirus, transfected into HEK293 cells, expanded, isolated by immobilized metal affinity chromatography (IMAC), cleaved with tobacco etch virus protease, and purified to homogeneity by reverse-phase chromatography. Following purification, each peptide was lyophilized and stored frozen.

EXAMPLE 2

Radiolabeling of Peptide

[0210] This example describes radiolabeling of peptides with standard techniques. See J Biol Chem. 254(11):4359-65 (1979). The sequences were engineered to have the amino acids, "G" and "S" at the N terminus. See Methods in Enzymology V91: 1983 p. 570 and Journal of

Biological Chemistry 254(11): 1979 p. 4359. An excess of formaldehyde was used to ensure complete methylation (dimethylation of every free amine). The labeled peptides were isolated via solid-phase extraction on Strata-X columns (Phenomenex 8B-S 100-AAK), rinsed with water with 5% methanol, and recovered in methanol with 2% formic acid. Solvent was subsequently removed in a blowdown evaporator with gentle heat and a stream of nitrogen gas. EXAMPLE 3

Peptide Detectable Agent Conjugates

[0211] This example describes the dye labeling of peptides. A peptide of the disclosure is expressed recombinantly or chemically synthesized, and then the N-terminus of the peptide is conjugated to a detectable agent via an NHS ester using DCC or EDC to produce a peptide- detectable agent conjugate. The detectable agent is the fluorophore dye is a cyanine dye, such as Cy5.5 or an Alexa fluorophore, such as Alexa647.

[0212] The peptide detectable agent conjugates are administered to a subject. The subject can be a human or a non-human animal. After administration, the peptide detectable agent conjugates home to cartilage. The subject, or a biopsy from the subject, can be imaged to visualize localization of the peptide detectable agent conjugates to cartilage. In some aspects, visualization of the peptide detectable agent conjugates in cartilage after administration results in diagnosis of arthritis, cartilage damage, or any cartilage disorder.

EXAMPLE 4

Method to Determine Improved Peptide Variants

[0213] This example shows a method for determining ways to improve peptide variants by comparing and analyzing the primary sequences and tertiary structures of scaffold peptides. FIG. 5A-FIG. 5C show sequences of SEQ ID NO: 541 aligned with SEQ ID NO: 316, SEQ ID NO: 541 aligned with SEQ ID NO: 542, and SEQ ID NO: 541 aligned with SEQ ID NO: 483. The sequence alignment of the two scaffolds was used to identify conserved positively charged residues (shown in boxes) that may be important for cartilage homing. A peptide of SEQ ID NO: 483 homes to cartilage and other peptides with positively charged residues in similar positions, or cysteines in similar positions, or other residues that are in similar positions are also predicted to home to cartilage.

[0214] FIG. 6 shows sequences of SEQ ID NO: 320 aligned with SEQ ID NO: 484. The sequence alignment of the two scaffolds was used to identify the basic/aromatic dyad that may be involved in the interaction with the Kv ion channel (K27 and Y36 of SEQ ID NO: 484). The mutation of K27 to alanine, arginine, or glutamic acid destroyed activity against the squid KvlA ion channel. K27 and Y36 may be desirable to maintain or add to a cartilage homing peptide of this disclosure to maintain or improve homing, to maintain or improve residence time in cartilage, or to maintain or improve modulation of an ion channel such as Kv. In contrast, K27 and Y36 may be desirable to mutate out of a cartilage homing peptide to reduce interaction with an ion channel such as Kv. Disruption of either the basic or aromatic residue eliminates ion channel activity. In another example, D amino acids are expected to reduce or eliminate binding. EXAMPLE 5

Sequence Alignment to pFam00451:toxin_2 Family to Identify Cartilage Homing Peptides

[0215] This example describes a method for identifying new cartilage homing peptides by sequence alignment to the pFam00451:toxin_2 structural class family. The pFam00451:toxin_2 structural class is a family of peptides related by similarities in sequence identity. FIG. 7

illustrates alignment of peptides within the pfam00451:toxin_2 structural class family of SEQ ID NO: 494 - SEQ ID NO: 540. Boxed and bolded residues indicate relative conservation of sequence while non-boxed and non-bo lded residues indicate areas of higher sequence variability. SEQ ID NO: 494 was identified as a cartilage homing candidate peptide based on its structural similarities with the pFam00451:toxin_structural class family. FIG. 8 illustrates the sequence alignment of a peptide of SEQ ID NO: 494 from the pfam00451:toxin 2 structural class family with the sequence of SEQ ID NO: 27. Asterisks indicate positions with a single, fully conserved residue, a colon indicates conservation between groups of strongly similar properties (scoring > 0.5 in the Gonnet point accepted mutation (PAM) 250 matrix), and a period indicates

conservation between groups of weakly similar properties (scoring < 0.5 in the Gonnet PAM 250 matrix). SEQ ID NO: 108 was also identified as a cartilage homing candidate based on its structural similarities with the pfam00451:toxin_2 structural class family of peptides.

[0216] The pFam00451:toxin_2 structural class family is used as a scaffold to identify variant peptides that have cartilage homing properties. Any member of the pFam00451:toxin_2 structural class family is used to predict new cartilage homing peptides based on homology, preserved residues, or a preserved cysteine residue.

EXAMPLE 6

Dosing of Peptide with Kidney Ligation

[0217] This example describes a dosing scheme for administering peptides to mice in

conjunction with kidney ligation. Different dosages of the peptides were administered to Female Harlan athymic nude mice, weighing 20g - 25g, via tail vein injection (n = 2 mice per peptide). The sequence of thirteen cartilage homing peptides of SEQ ID NO: 24 - SEQ ID NO: 36 are shown in TABLE 1. The experiment was done in duplicates. The kidneys were ligated to prevent renal filtration of the peptides. Each peptide was radiolabeled by methylating lysines and the N- terminus, so the actual binding agent may contain methyl or dimethyl lysine(s) and a methylated or dimethylated amino terminus.

[0218] A target dosage of 50-100 nmol of each peptide carrying 10-25 uCi of ¹⁴ C was administered to Female Harlan athymic nude mice while anesthetized. Each peptide was allowed to freely circulate within the animal before the animals were euthanized and sectioned. EXAMPLE 7

Peptide Homing with Kidney Ligation

[0219] This example illustrates peptide homing to cartilage of mice with kidneys that were ligated prior to peptide administration. At the end of the dosing period in EXAMPLE 6, mice were frozen in a hexane/dry ice bath and then frozen in a block of carboxymethylcellulose.

Whole animal sagittal slices were prepared that resulted in thin frozen sections being available for imaging. Thin, frozen sections of animal including imaging of tissues such as brain, tumor, liver, kidney, lung, heart, spleen, pancreas, muscle, adipose, gall bladder, upper gastrointestinal tract, lower gastrointestinal tract, bone, bone marrow, reproductive track, eye, cartilage, stomach, skin, spinal cord, bladder, salivary gland, and other types of tissues were obtained with a microtome, allowed to desiccate in a freezer, and exposed to phosphoimager plates for about ten days.

[0220] These plates were developed, and the signal (densitometry) from each organ was normalized to the signal found in the heart blood of each animal. A signal in tissue darker than the signal expected from blood in that tissue indicates peptide accumulation in a region, tissue, structure or cell. For instance, the cartilage is avascular and contains minute amounts of blood. A ratio of at least 170% signal in the cartilage versus heart ventricle was chosen as a reference level for significant targeting to cartilage, which also correlated with clear accumulation in

cartilaginous tissues in the images of the slices. FIG. 1 identifies the locations of the SEQ ID NO: 27 peptide distribution in joint and other cartilage. FIG. 12 identifies the locations of the SEQ ID NO: 27 peptide distribution in nasal, spinal, tracheal, and other cartilage, including to hyaline cartilage such as articular cartilage and physeal cartilage, as well as fibrocartilage.

[0221] Additionally, the peptide can be retained in cartilage for hours after treatment. The SEQ ID NO: 27 peptide was radiolabeled as in EXAMPLE 6 and 100 nmol of peptide was injected into a mouse with intact kidneys. FIG. 9 illustrates the retention of and the tissue distribution in the cartilage of a peptide of SEQ ID NO: 27, 24 hours after administration.

EXAMPLE 8

Dosing of Peptide without Kidney Ligation

[0222] This example describes a dosing scheme for administering peptides to mice without kidney ligation. The peptide administered had the sequence of SEQ ID NO: 27 as shown in

TABLE 1. The peptide was radiolabeled by methylating lysines and the N-terminus, so the actual binding agent may contain methyl or dimethyl lysine(s) and a methylated or dimethylated amino terminus.

-I l l- [0223] A target dosage of 100 nmol of each peptide carrying 10-25 μθ of ¹⁴ C was administered to Female Harlan athymic nude mice by a tail vein injection. Each peptide was allowed to freely circulate within the animal for either 4 hours or 24 hours before the animals were euthanized and sectioned.

EXAMPLE 9

Peptide Homing with Intact Kidneys

[0224] This example illustrates peptide homing to cartilage in animals with intact kidneys. At the end of the 4 hour or 24 hour dosing periods in EXAMPLE 8, mice were frozen in a hexane/dry ice bath and then frozen in a block of carboxymethylcellulose. Whole animal sagittal slices were prepared that resulted in thin frozen sections being available for imaging. Thin, frozen sections of animal including imaging of tissues such as brain, tumor, liver, kidney, lung, heart, spleen, pancreas, muscle, adipose, gall bladder, upper gastrointestinal track, lower gastrointestinal track, bone, bone marrow, reproductive track, eye, cartilage, stomach, skin, spinal cord, bladder, salivary gland, and other types of tissues were obtained with a microtome, allowed to desiccate in a freezer, and exposed to phospho imager plates for about ten days.

[0225] These plates were developed. A signal in tissue darker than the signal expected from blood in that tissue indicates peptide accumulation in a region, tissue, structure or cell. For instance, the cartilage is avascular and contains minute amounts of blood. High signal in the kidneys indicates presence and accumulation of the peptide in the kidneys. FIG. 1 identifies the locations of the SEQ ID NO: 27 peptide distribution in joint and other cartilage as well as kidneys.

EXAMPLE 10

Peptide Homing with Therapeutic Agents

[0226] This example describes certain exemplary therapeutic agents that are conjugated to a peptide. A peptide of the disclosure is expressed recombinantly or chemically synthesized and then is conjugated to an exemplary drug, such as paclitaxel or triamcinolone acetonide or budesonide using techniques known in the art, such as those described in Bioconjugate

Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd Edition, 2013). One or more drugs is conjugated per peptide, or an average of less than one drug is conjugated per peptide.

[0227] Coupling of these drugs to a peptide of any of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564 targets the drug to the cartilage of the subject. One or more drug-peptide conjugates are administered to a human or animal. EXAMPLE 11

Peptide Homing to an Arthritic Joint

[0228] This example illustrates peptide homing to cartilage in humans or animals with arthritis. A peptide of the present disclosure is expressed recombinantly or chemically synthesized and is used directly, after radiolabeling, or after conjugation to a fluorophore or therapeutic compound. A peptide is selected from any one of the peptides of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. The peptide or peptide conjugate is administered to a human or animal subcutaneously, intravenously, or orally, or is injected directly into a joint intraarticularly. The peptide or peptide conjugate homes to cartilage.

EXAMPLE 12

Peptide Homing to Cartilage in Non-Human Animals

[0229] This example illustrates a peptide or peptide conjugate of this disclosure homing to cartilage in non-human animals. Non-human animals include but are not limited to guinea pigs, rabbits, dog, cats, horses, rats, mice, cows, pigs, non-human primates, and other non-human animals. A peptide of the present disclosure is recombinantly expressed or chemically

synthesized and are used directly, after radiolabeling, or after conjugation to a fluorophore or therapeutic compound. The peptide is selected from any one of the peptides of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. The resulting peptide or peptide conjugate is administered to a non-human animal subcutaneously, intravenously, or orally, or is injected directly into a joint intra- articularly. Biodistribution is assessed by LC/MS,

autoradiography, positron emission tomography (PET), or fluorescence imaging. A peptide or peptide conjugate is homed to cartilage in non-human animals.

EXAMPLE 13

Whole Body Fluorescence and Isolated Limb Fluorescence of Homing Peptides

[0230] This example illustrates whole body fluorescence and isolated limb fluorescence of peptide homers of this disclosure. Any peptide of the present disclosure is chemically conjugated to one molecule of a near infrared fluorophore, at the N-terminus of the peptide via an active NHS ester on the dye. A dose of 10 nmol of each peptide conjugated to a fluorophore is administered to Female Harlan athymic nude mice, weighing 20-25 g, and is administered via tail vein injection. Each experiment is done at least in duplicate (n=2 mice per group). The peptide fluorophore conjugate is allowed to freely circulate for the described time period before the mice were euthanized at various time points. Mice are evaluated for peptide distribution of the peptide fluorescence in whole body imaging and in isolated hind limb imaging. [0231] For Whole body fluorescence (WBF), at the end of the dosing period, mice are frozen in a hexane/dry ice bath and then embedded in a frozen block of carboxymethylcellulose. Whole animal sagittal slices are prepared that resulted in thin frozen sections for imaging. Thin frozen sections are obtained using a microtome and allowed visualization of tissues. Sections are allowed to dessicate in a freezer prior to imaging. WBF is performed on fluorescent sections, which are scanned on a Li-Cor Odyssey scanner at a setting of 169 μιη resolution, medium quality, 700 channel, L-2.0 intensity.

[0232] For isolated hind limb fluorescence studies, mice are euthanized by C0 ₂ asphyxiation at the end of the dosing period. The right hind limb is removed at the hip joint and imaged on a Sepctrum IVIS imager (ex/em: 675 nm. 720 nm) with a 1 second exposure length and a focal height of 0.5 cm. Limbs are imaged with skin removed and with muscle removed.

EXAMPLE 14

Whole Body Autoradiography of Homing Peptides

[0233] This example illustrates whole body autoradiography of peptide homers of this disclosure. Peptides are radiolabeled by methylating lysines at the N-terminus as described in EXAMPLE 2. As such, the peptide may contain methyl or dimethyl lysines and a methylated or dimethlyated amino terminus. A dose of 100 nmol radiolabeled peptide is administered via tail vein injection in Female Harlan athymic nude mice, weighing 20-25 g. The experiment is done in at least duplicate (n=2 animals per group). In some animals, kidneys are ligated to prevent renal filtration of the radio labled peptides and extend plasma half-life. Each radiolabeled peptide is allowed to freely circulate within the animal for the described time period before the animals were euthanized and sectioned.

[0234] Whole body autoradiography (WBA) sagittal sectioning is performed as follows. At the end of the dosing period, mice are frozen in a hexane/dry ice bath and then embedded in a frozen block of carboxymethylcellulose. Whole animal sagittal slices are prepared that resulted in thin frozen sections for imaging. Thin frozen sections are obtained using a microtome and allowed visualization of tissues such as brain, tumor, liver, kidney, lung, heart, spleen, pancreas, muscle, adipose, gall bladder, upper gastrointestinal tract, lower gastrointestinal tract, bone, bone marrow, reproductive tract, eye, cartilage, stomach, skin, spinal cord, bladder, salivary gland, and more. Sections are allowed to dessicate in a freezer prior to imaging.

[0235] For the autoradiography imaging, tape mounted thin sections are freeze dried and radioactive samples were exposed to phosphoimager plates for 7 days. These plates are developed and the signal (densitometry) from each organ was normalized to the signal found in the cardiac blood of each animal. A signal in tissue darker than the signal expected from blood in that tissue indicates accumulation in a region, tissue, structure, or cell.

EXAMPLE 15

Peptide Localization in Chondrocytes

[0236] This example illustrates binding of peptides of this disclosure to chondrocytes within cartilage in animals with intact kidneys. In one embodiment, animals are dosed and are processed as described in EXAMPLE 13 and EXAMPLE 14. At the end of the dosing period, animals are euthanized and cartilage is optionally removed for use in staining and imaging procedures. Whole animal sagittal slices are prepared that result in thin frozen sections being available for staining and imaging. One or more of the following cartilage components are identified in thin frozen sections or live cartilage explants using standard staining techniques: collagen fibrils, glycosaminoglycans, or chondrocytes. A peptide of this disclosure is found to localize to chondrocytes in cartilage, localized intracellularly or extracellularly bound or both. Localization is visualized and confirmed by microscopy.

[0237] In another embodiment, peptides or peptide-drug conjugates of this disclosure are administered in humans and are localized on or in chondrocytes in cartilage.

EXAMPLE 16

Peptide Localization in Cartilage Extracellular Matrix

[0238] This example illustrates localization of peptides of this disclosure in cartilage

extracellular matrix. In one embodiment, animals are dosed and are processed as described in EXAMPLE 13 and EXAMPLE 14 in animals with intact kidneys. At the end of the dosing period, animals are euthanized and cartilage is optionally removed for use in staining and imaging procedures. Whole animal sagittal slices are prepared that result in thin frozen sections being available for staining and imaging. Thin frozen sections or live cartilage explants are acquired, stained, and visualized as described in EXAMPLE 15. A peptide of the present disclosure is found to localize to the extracellular matrix in cartilage. The peptide may be bound to one or more components of the extracellular matrix, such as proteoglycans,

glycosaminoglycans, aggrecan, decorin, or collagen. Localization is visualized and confirmed by microscopy.

[0239] In another embodiment, peptides or peptide-drug conjugates of this disclosure are administered in humans and are localized in cartilage extracellular matrix. EXAMPLE 17

Peptide Binding to Cartilage Explants

[0240] This example illustrates a peptide or peptide conjugation of this disclosure homing, targeting, being directed to, migrating to, being retained by, accumulating in, or binding to human and animal cartilage explants in culture. A peptide is selected from any one of the peptides of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Peptides are recombinantly expressed or chemically synthesized and are used directly, after radiolabeling, or after conjugation to a fluorophore or therapeutic compound. A peptide of peptide conjugate of this disclosure is incubated with cartilage explants derived from humans or animals. Peptides of peptide conjugate are found to bind to cartilage explants. The interaction with cartilage is confirmed using various methods that include but are not limited to liquid scintillation counting, confocal microscopy, immunohistochemistry, HPLC, or LC/MS. The peptide shows a higher level of signal than a control peptide that is administered that is not a cartilage binding peptide.

EXAMPLE 18

Effects of Peptide on Ion Channels

[0241] This example describes the interaction between peptides of the present disclosure and ion channels. Ion channels can be associated with pain and can be activated in disease states such as arthritis. A peptide of the disclosure is expressed and administered in a pharmaceutical composition to a patient to treat a joint condition or disease associated with an ion channel and treatable by binding, blocking, or interacting with the ion channel. Ion channels, such as Nav 1. 7, are inhibited by peptides of the present disclosure. A given peptide is expressed recombinantly or chemically synthesized, wherein the peptide selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Following expression or synthesis, the peptide is used directly or conjugated to a therapeutic compound, such as those described herein. A peptide of the present disclosure selectively interacts with ion channels, or is mutated in order to interact with ion channels. For example, a peptide of this disclosure is bound to Nav 1. 7 or Nav 1. 7 is blocked by a peptide of this disclosure. When the peptide is administered to a human subject, Nav 1.7 signaling is reduced in the tissues in proximity to the joints, and pain relief is thereby provided.

EXAMPLE 19

Peptide-Fc Protein Fusions

[0242] This example illustrates making and using peptide-Fc protein fusions. A peptide of SEQ ID NO: 108 was recombinantly expressed with the sequence for the human IgGl Fc protein in HEK293 cells to yield a sequence of SEQ ID NO: 565

(METDTLLLWVLLLW VPGS TGGS G VPIN VRCRGS RDCLDPCRR AGMRFGRCINS RCHCT PGGS GGS DKTHTCPPCP APELLGGPS VFLFPPK KDTLMIS RTPE VTC V V VD VS HEDPE V KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP APIEKTIS KAKGQPREPQ V YTLPPS RDELTKNQ VS LTCLVKGFYPS DI A VEWES NGQPEN NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK).

[0243] The sequence of any peptide of this disclosure is expressed as a fusion protein with either murine or human Fc by adding a secretion signal sequence to the N-terminus and an Fc sequence to the C-terminus. This creates a bivalent molecule with improved secretion properties. The larger peptide-Fc fusion is expressed in different mammalian or insect cell lines and is useful as a research reagent and a therapeutic.

[0244] Fc fusion to a peptide of SEQ ID NO: 108 to yield a sequence of SEQ ID NO: 565 extends half-life and improves biodistribution of the peptide to cartilage. Any peptide of this disclosure is co-expressed with Fc protein to yield Fc-fusion peptides with longer half-life and improved homing to cartilage. In SEQ ID NO: 565, the secretion signal sequence

METDTLLLWVLLLWVPGSTG (SEQ ID NO: 566) is followed by the peptide of SEQ ID NO: 108, and is followed by the sequence for Fc protein. Cleaving can be imprecise, resulting in cleavage at position 20 or position 21 of SEQ ID NO: 565.

EXAMPLE 20

Peptide Conjugate Hydrolysis

[0245] This example describes preparation of peptide conjugates having tunable hydrolysis rates. The peptide-drug conjugates described below are synthesized with the modification that instead of using succinic anhydride, other molecules are used to provide steric hindrance to hydrolysis or an altered local environment at the carbon adjacent to the final hydrolyzable ester. In one exemplary conjugate, the peptide-drug conjugate is synthesized with tetramethyl succinic anhydride to generate hindered esters, which causes a decreased rate of hydrolysis. In another exemplary conjugate, one methyl group is present at the adjacent carbon. In another exemplary conjugate, two methyl groups are present at the adjacent carbon. In another exemplary conjugate, one ethyl group is present at the adjacent carbon. In another exemplary conjugate, two ethyl groups are present at the adjacent carbon. In another exemplary conjugate, the carbon linker length is increased such as by using glutaric anhydride instead of succinic anhydride, increasing the local hydrophobicity and lowering the hydrolysis rate. In another exemplary conjugate, a hydro xyl group is located on the adjacent carbon, increasing the local hydrophilicity and increasing the hydrolysis rate. The rate of hydrolysis in these exemplary conjugates is therefore adjusted, preventing premature cleavage and ensuring that the majority of peptide- dexamethasone conjugates accumulate in cartilage prior to release of the drug by hydrolysis but that the dexamethasone is also released in the cartilage in a timely manner.

[0246] The resulting peptide conjugates are administered to a human or animal subcutaneously, intravenously, orally, or injected directly into a joint to treat disease.

EXAMPLE 21

Peptide Conjugates with Stable Linkers

[0247] This example describes preparation of peptide conjugates with stable linkers. A peptide of the disclosure is expressed recombinantly or is chemically synthesized. The peptide is conjugated to a detectable agent or an active agent via a stable linker, such as an amide linkage or a carbamate linkage. The peptide is conjugated to a detectable agent or an active agent via a stable linker, such as an amide bond using standard l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) or dicylcohexylcarbodiimide (DCC) based chemistry or thionyl chloride or phosphorous chloride-based bioconjugation chemistries.

[0248] A peptide and drug conjugated via a linker are described with the formula Peptide- A-B- C-Drug, wherein the linker is A-B-C. A can be a stable amide link that is formed by reacting with an amine on the peptide with a linker containing a tetrafluorophenyl (TFP) ester or an NHS ester. A can also be a stable carbamate linker that is formed by reacting with an amine on the peptide imidazole carbamate active intermediate formed by the reaction of CDI with a hydroxyl on the linker. A can also be a stable secondary amine linkage that is formed by reductive alkylation of the amine on the peptide with an aldehyde or ketone group on the linker. A can also be a stable thioether linker formed using a maleimide or bromoacetamide in the linker with a thiol in the peptide, a triazole linker, a stable oxime linker, or an oxacarboline linker. B is (-CH2-) _X -, a short PEG (-CH ₂ CH ₂ 0-) _x (x is 0-20). Alternatively, spacers within the linker is optional and can be included or not at all. C is an amide bond formed with an amine or a carboxylic acid on the drug, a thioether formed between a maleimide on the linker and a sulfhydroyl on the drug, a secondary or tertiary amine, a carbamate, or other stable bonds. Any linker chemistry described in "Current ADC Linker Chemistry," Jain et al., Pharm Res, 2015 DOI 10. 1007/sl 1095-015-1657-7 can be used.

[0249] The resulting peptide conjugates are administered to a human or animal subcutaneously, intravenously, orally, or injected directly into a joint to treat disease. The peptide is not specifically cleaved from the detectable agent or active agent via a targeted mechanism. The peptide can be degraded by mechanisms such as catabolism, releasing a drug that is modified or not modified form its native form (Singh, Luisi, and Pak, Pharm Res 32:3541-3571 (2015)). The peptide drug conjugate exerts its pharmacological activity while still intact, or while partially or fully degraded, metabolized, or catabolized.

EXAMPLE 22

Peptide Conjugates with Cleavable Linkers

[0250] This example describes preparation of peptide conjugates having cleavable linkers. A peptide of the disclosure is expressed recombinantly or chemically synthesized. A peptide and drug are conjugated via a linker and is described with the formula Peptide- A-B-C-Drug, wherein the linker is A-B-C. A is a stable amide link such as that formed by reacting an amine on the peptide with a linker containing a tetrafluorophenyl (TFP) ester or an NHS ester. A can also be a stable carbamate linker such as that formed by reacting an amine on the peptide with an imidazole carbamate active intermediate formed by reaction of CDI with a hydroxyl on the linker. A can also be a stable secondary amine linkage such as that formed by reductive alkylation of the amine on the peptide with an aldehyde or ketone group on the linker. A can also be a stable thioether linker formed using a maleimide or bromoacetamide in the linker with a thiol in the peptide, a triazole linker, a stable oxime linker, or a oxacarboline linker. B is (-CH2- ) _x - or a short PEG (-Ο¾Ο-ΐ20-) _χ (x is 0-20) or other spacers or no spacer. C is an ester bond to the hydroxyl or carboxylic acid on the drug, or a carbonate, hydrazone, or acylhydrazone, designed for hydro lytic cleavage. The hydro lytic rate of cleavage is varied by varying the local environment around the ester, including carbon length (-CH2-)x, steric hindrance (including adjacent side groups such as methyl, ethyl, cyclic), hydrophilicity or hydrophobicity. Hydrolysis rate is affected by local pH, such as lower pH in certain compartments of the body or of the cell such as endosomes and lysosomes or diseased tissues. C is a pH sensitive group such as a hydrazone or oxime linkage. Alternatively C is a disulfide bond designed to be released by reduction, such as by glutathione. Alternatively C (or A-B-C) is a peptidic linkage design for cleavabe by enzymes. Optionally, a self- immolating group such as pABC is included to cause release of a free unmodified drug upon cleavage (Antibody- Drug Conjugates: Design,

Formulation, and Physicochemical Stability, Singh, Luisi, and Pak. Pharm Res (2015) 32:3541- 3571). The linker is cleaved by enzymes such as esterases, matrix metalloproteinases, cathepsins such as cathepsin B, glucuronidases, a protease, or thrombin. Alternatively, the bond designed for cleavage is at A, rather than C, and C could be a stable bond or a cleavable bond. An alternative design is to have stable linkers (such as amide or carbamate) at A and C and have a cleavable linker in B, such as a disulfide bond. The rate of reduction is modulated by local effects such as steric hindrance from methyl or ethyl groups or modulating hydrophobicity/hydrophilicity. [0251] The resulting peptide conjugates are administered to a human or animal subcutaneously, intravenously, orally, or injected directly into a joint to treat disease.

EXAMPLE 23

Acetylsalicylic Acid Peptide Conjugate

[0252] This example describes the conjugation of acetylsalicylic acid to a peptide using a lactic acid linker. A conjugate is produced from a mixture of (R,S)- acetylsalicylic acid, lactic acid, and a peptide:

[0253] The acetylsalicylic acid -lactic acid linker conjugate depicted above is then reacted with a lysine or the N-terminus of a cystine-dense peptide to create an acetylsalicylic acid -lactic acid- peptide conjugate. The cystine-dense peptide is selected from the peptides of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564.

[0254] Acetylsalicylic acid is currently dosed as an enantiomeric mixture, in which enantiomers with a single racemic stereocenter are very difficult to separate. As in the reaction scheme (I), a diastereomer with two chiral centers is created by the addition of a chiral linker such as L- lactic acid. Since diastereomers are easily separated, the active enantiomer of acetylsalicylic acid conjugated to the lactic acid linker can be purified prior to conjugation to a cystine-dense peptide. The chemical synthesis can use any conjugation techniques known in the art, such as described in Bioconjugate Techniques by Greg Hermanson and in "Ketorolac-dextran conjugates: synthesis, in vitro, and in vivo evaluation:" Acta Pharm. 57 (2007) 441-450, Vyas, Trivedi, and Chaturvedi. The conjugate can display anti- inflammatory activity, or free acetylsalicylic acid is released from the conjugate to provide anti- inflammatory activity. The free acetylsalicylic acid can result from hydrolysis that occurs after administration, such as hydrolysis at the ester bond. By dosing the conjugate containing the cartilage homing peptide, a higher AUC of acetylsalicylic acid delivery to the joint may be achieved than would be achieved by systemic dosing of acetylsalicylic acid alone.

[0255] Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e. g., EXAMPLES 21 and 22).

EXAMPLE 24

Acetylsalicylic Acid Peptide Conjugate

[0256] This example describes the conjugation of acetylsalicylic acid to a peptide using a PEG linker. A conjugate is produced using acetylsalicylic acid and a PEG linker, which forms an ester bond that can hydrolyze as described in "In vitro and in vivo study of poly(ethylene glycol) conjugated ibuprofen to extend the duration of action," Scientia Pharmaceutica, 2011, 79:359- 373, Nayak and Jain. Fischer esterification is used to conjugate ibuprofen with a short PEG, e.g., with triethylene glycol, to yield ibuprofen-ester-PEG-OH.

[0257] Following preparation of the PEG-ibuprofen conjugate as shown above, the hydroxyl moiety of PEG is activated with Ν,Ν'-disuccinimidyl carbonate (DSC) to form ibuprofen-ester- PEG-succinimidyl carbonate, which is then reacted with a lysine or the N-terminus of a cystine- dense peptide to form an ibuprofen-ester-PEG-peptide conjugate. The cystine-dense peptide is selected from any one of the peptides of sequence SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. The conjugate can display anti- inflammatory activity, or free ibuprofen is released from the conjugate to provide anti- inflammatory activity. The free ibuprofen can result from hydrolysis that occurs after administration, such as hydrolysis at the ester bond.

[0258] Ibuprofen-peptide conjugates are administered to a subject in need thereof. The subject can be a human or a non-human animal.

[0259] Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 25

Dexamethasone Peptide Conjugate

[0260] This example describes different methods of conjugating dexamethasone with a peptide of this disclosure. A peptide of SEQ ID NO: 108 was recombinantly expressed. Dexamethasone was readily conjugated to a peptide of this disclosure using a dicarboxylic acid linker. The peptide-dexamethasone conjugate was made by first converting dexamethasone to a

hemisuccinate by reacting it with succinic anhydride. The hemisuccinate was then converted to a succinate carboxylic acid containing an active ester, using dicyclohexyl carbodiimide (DCC) or l-ethyl-3-(3-dimethylamninopropyl)carbodiimide (EDC) in the presence of N-hydroxy succinimide (NHS). This active ester was then reacted with a lysine or the N-terminus of a cystine-dense peptide to create a dexamethasone-carboxylic acid-peptide conjugate. Methods such as those described in "Functionalized derivatives of hyaluronic acid oligosaccharides: drug carriers and novel biomaterials" Bioconjugate Chemistry 1994, 5, 339-347, Pouyani and

Prestwich, and Bioconjugate Techniques by Greg Hermanson can be used (Elsevier Inc., 3 ^rd Edition, 2013).

[0261] Peptide-dexamethasone conjugates were prepared by coupling dexamethasone to the peptides of this disclosure using standard coupling -reagent chemistry. For example,

dexamethasone conjugates were made by reacting dexamethasone hemigluterate with 1.05 molar equivalents of Ι, -carbonyldiimidazole in anhydrous DMSO in an inert atmosphere. After 30 minutes, excess dexamethasone in anhydrous DMSO was added along with two molar equivalents of anhydrous trimethylamine. The N-hydroxysuccinimide ester of the peptide- dexamethasone conjugate was generated to form a shelf- stable intermediate for later reaction with an amine-containing carrier. The N-terminal dexamethasone-peptide conjugate (SEQ ID NO: 108B) was verified by electrospray mass spectrometry (ES-MS) within a 10 ppm error.

[0262] A peptide of any of the sequences of this disclosure including SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564, are conjugated to dexamethasone using the methods described above.

[0263] Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 26

Beclomethasone monopropionate Peptide Conjugates

[0264] This example describes conjugation of a peptide of SEQ ID NO: 27 or SEQ ID NO: 108 of this disclosure to beclomethasone monopropionate. Beclomethasone monopropionate is readily conjugated to any peptide disclosed herein via a dicarboxylic acid linker. The

dicarboxylic acid linker is a linear dicarboxylic acid, such as succinic acid, or a related cyclic anhydride, such as succinic anhydride. Reactions with anhydrides can proceed under simple conditions. For example, the reaction of beclomethasone monopropionate with five molar equivalents of glutaric anhydride is carried out in anhydrous pyridine at room temperature.

Reactions with dicarboxylic acids can occur using standard carbodiimide coupling methods. For example, beclomethasone monopropionate is reacted with one molar equivalent dimethylsuccinic acid, one molar equivalent l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (or another carbodiimide), and 0.2 molar equivalents of 40-dimethylamino pyridine.

[0265] The same methods as described in EXAMPLE 20 are used to adjust the rate of hydrolysis of peptide- beclomethasone monopropionate conjugates, preventing premature cleavage and ensuring that the beclomethasone monopropionate of peptide- beclomethasone monopropionate conjugates accumulate in cartilage.

[0266] Peptide- beclomethasone monopropionate conjugates are prepared by coupling beclomethasone monopropionate to the peptides of this disclosure using standard coupling- reagent chemistry. The peptide- beclomethasone monopropionate conjugate was made by first converting beclomethasone monopropionate to a hemisuccinate by reacting it with succinic anhydride. The hemisuccinate was then converted to a succinate carboxylic acid containing an active ester, using dicyclohexyl carbodiimide (DCC) or l-ethyl-3-(3- dimethylamninopropyl)carbodiimide (EDC) in the presence of N-hydroxy succinimide (NHS). This active ester was then reacted with a lysine or the N-terminus of a peptide to create a beclomethasone monopropionate -carboxylic acid-peptide conjugate. Methods such as those described in "Functionalized derivatives of hyaluronic acid oligosaccharides: drug carriers and novel biomaterials" Bioconjugate Chemistry 1994, 5, 339-347, Pouyani and Prestwich, and Bioconjugate Techniques by Greg Hermanson can be used (Elsevier Inc., 3 ^rd Edition, 2013).

[0267] Peptide- beclomethasone monopropionate conjugates were prepared by coupling beclomethasone monopropionate to the peptides of this disclosure using standard coupling- reagent chemistry. For example, beclomethasone monopropionate conjugates were made by reacting beclomethasone monopropionate hemigluterate with 1.05 molar equivalents of 1,1'- carbonyldiimidazole in anhydrous DMSO in an inert atmosphere. After 30 minutes, excess beclomethasone monopropionate in anhydrous DMSO was added along with two molar equivalents of anhydrous trimethylamine. The N-hydroxysuccinimide ester of the peptide- beclomethasone monopropionate conjugate was generated to form a shelf- stable intermediate for later reaction with an amine-containing carrier.

[0268] Beclomethasone monopropionate is also readily conjugated to any peptide disclosed herein via a dicarboxylic acid linker. The dicarboxylic acid linker is a linear dicarboxylic acid, such as succinic acid, or a related cyclic anhydride, such as succinic anhydride. Reactions with anhydrides can proceed under simple conditions. For example, the reaction of beclomethasone monopropionate with five molar equivalents of glutaric anhydride is carried out in anhydrous pyridine at room temperature. Reactions with dicarboxylic acids can occur using standard carbodiimide coupling methods. For example, beclomethasone monopropionate is reacted with one molar equivalent dimethylsuccinic acid, one molar equivalent l-ethyl-3-(3- dimethylaminopropyl)carbodiimide (or another carbodiimide), and 0.2 molar equivalents of 40- dimethylamino pyridine. The peptide- beclomethasone monopropionate conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage and/or kidneys. The subject is a human or animal and has inflammation in the cartilage or kidney tissues. Upon administration of the peptide- beclomethasone monopropionate conjugates, the cartilage and/or kidney inflammation is alleviated.

[0269] The peptide can also be a peptide of SEQ ID: NO: 33. The peptide can be any peptide with the sequence selected SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564.

[0270] Such peptide-drug conjugates are made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22). EXAMPLE 27

Desciclesonide Peptide Conjugates

[0271] This example describes conjugation of a peptide of SEQ ID NO: 199 or SEQ ID NO: 187 of this disclosure to desciclesonide. Ciclesonide is a prodrug that is metabolized in vivo to the active metabolite desciclesonide. By conjugating desciclesonide to a peptide via an ester linker, upon hydrolysis the released drug would be desciclesonide, just as after systemic administration of ciclesonide the active metabolite desciclesonide is present and active. Desciclesonide is readily conjugated to any peptide disclosed herein via a dicarboxylic acid linker. The

dicarboxylic acid linker is a linear dicarboxylic acid, such as succinic acid, or a related cyclic anhydride, such as succinic anhydride. Reactions with anhydrides can proceed under simple conditions. For example, the reaction of desciclesonide with five molar equivalents of glutaric anhydride is carried out in anhydrous pyridine at room temperature. Reactions with dicarboxylic acids can occur using standard carbodiimide coupling methods. For example, desciclesonide is reacted with one molar equivalent dimethylsuccinic acid, one molar equivalent l-ethyl-3-(3- dimethylaminopropyl)carbodiimide (or another carbodiimide), and 0.2 molar equivalents of 40- dimethylamino pyridine.

[0272] The same methods as described in EXAMPLE 20 are used to adjust the rate of hydrolysis of peptide- desciclesonide conjugates, preventing premature cleavage and ensuring that the desciclesonide of peptide- desciclesonide conjugates accumulate in cartilage.

[0273] Desciclesonide is also readily conjugated to any peptide disclosed herein via a dicarboxylic acid linker. The dicarboxylic acid linker is a linear dicarboxylic acid, such as succinic acid, or a related cyclic anhydride, such as succinic anhydride. Reactions with anhydrides can proceed under simple conditions. For example, the reaction of desciclesonide with five molar equivalents of glutaric anhydride is carried out in anhydrous pyridine at room temperature. Reactions with dicarboxylic acids can occur using standard carbodiimide coupling methods. For example, desciclesonide is reacted with one molar equivalent dimethylsuccinic acid, one molar equivalent l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (or another carbodiimide), and 0.2 molar equivalents of 40-dimethylamino pyridine. The peptide- desciclesonide conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage and/or kidneys. The subject is a human or animal and has inflammation in the cartilage or kidney tissues. Upon administration of the peptide-desciclesonide conjugates, the cartilage and/or kidney inflammation is alleviated.

[0274] The peptide-desciclesonide conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage and/or kidneys. The subject is a human or animal and has inflammation in the cartilage or kidney tissues. Upon administration of the peptide-desciclesonide conjugates, the cartilage and/or kidney inflammation is alleviated.

[0275] The peptide can also be a peptide of SEQ ID: NO: 196. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564.

[0276] Such peptide-drug conjugates are made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 28

Desciclesonide Peptide Conjugates

[0277] This example describes conjugation of a peptide of SEQ ID NO: 199 or SEQ ID NO: 187 of this disclosure to desciclesonide. Ciclesonide is a prodrug that is metabolized in vivo to the active metabolite desciclesonide. By conjugating desciclesonide to a peptide via an ester linker, upon hydrolysis the released drug would be desciclesonide, just as after systemic administration of ciclesonide the active metabolite desciclesonide is present and active. Desciclesonide is readily conjugated to any peptide disclosed herein via a stable linker.

[0278] The peptide-desciclesonide conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage and/or kidneys. The subject is a human or animal and has inflammation in the cartilage or kidney tissues. Upon administration of the peptide-desciclesonide conjugates, the cartilage and/or kidney inflammation is alleviated.

[0279] The peptide can also be a peptide of SEQ ID: NO: 196. The peptide can be any peptide with the sequence selected SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564.

[0280] Such peptide-drug conjugates are made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 29

Peptide-Ustekinumab Conjugates

[0281] This example describes conjugation of a peptide of SEQ ID NO: 106 this disclosure to ustekinumab. Ustekinimab is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). Alternatively the peptide- active agent of this Example can be expressed as a fusion protein. From one to eight peptides are linked to ustekinumab.

[0282] The peptide-ustekinumab conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has psoriatic arthritis. Upon administration of the peptide- ustekinumab conjugates, the psoriatic arthritis condition is alleviated.

[0283] The peptide can also be a peptide of SEQ ID NO: 36. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564.

EXAMPLE 30

Peptide-Xeljanz Conjugates

[0284] This example describes conjugation of a peptide of SEQ ID NO: 187 this disclosure to xeljanz. Xeljanz is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to xeljanz.

[0285] The peptide- xeljanz conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has rheumatoid arthritis. Upon administration and homing of peptide- xeljanz conjugates, the rheumatoid arthritis condition is alleviated.

[0286] The peptide can also be a peptide of SEQ ID NO: 185. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 31

Peptide-IL-17 Inhibitor Conjugates

[0287] This example describes conjugation of a peptide of SEQ ID NO: 108 this disclosure to an IL-17 inhibitor. An IL-17 inhibitor is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013).

[0288] The peptide-IL-17 inhibitor conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has ankylosing spondylitis. Upon administration and homing of peptide-IL-17 inhibitor conjugates, the ankylosing spondylitis condition is alleviated. [0289] The peptide can also be a peptide of SEQ ID NO: 111. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 32

Peptide-Iguratimod Conjugates

[0290] This example describes conjugation of a peptide of SEQ ID NO: 199 this disclosure to iguratimod. Iguratimod is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013).

[0291] The peptide- iguratimod conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to kidneys. The subject is a human or animal and has rheumatoid arthritis. Upon administration and homing of peptide-iguratimod conjugates, the rheumatoid arthritis condition is alleviated.

[0292] The peptide can also be a peptide of SEQ ID NO: 26. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 33

Peptide Mycophenolic Acid Conjugates

[0293] This example describes conjugation of a peptide of SEQ ID NO: 27 this disclosure to mycophenolic acid. Mycophenolic acid is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013).

[0294] The peptide- mycophenolic acid conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to kidneys. The subject is a human or animal and has organ transplantation, infection, cancer, or other kidney disorders. Upon administration and homing of peptide- mycophenolic acid conjugates, the organ transplantation, infection, cancer, other kidney disorders condition is alleviated.

[0295] The peptide can also be a peptide of SEQ ID NO: 107. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 34

Peptide- Tacrolimus Conjugates

[0296] This example describes conjugation of a peptide of SEQ ID NO: 108 this disclosure to tacrolimus. Tacrolimus is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013)..

[0297] The peptide-tacrolimusconjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to kidneys. The subject is a human or animal and has organ transplantation, any other kidney disease. Upon administration and homing of peptide-tacrolimus conjugates, the organ transplantation, any other kidney disease condition is alleviated.

[0298] The peptide can also be a peptide of SEQ ID NO: 111. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 35

Peptide-Secukinumab Conjugates

[0299] This example describes conjugation of a peptide of SEQ ID NO: 106 this disclosure to secukinumab. Secukinumab is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to

secukinumab. Alternatively the peptide- active agent of this Example can be expressed as a fusion protein.

[0300] The peptide- secukinumab conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has ankylosing spondylitis. Upon administration and homing of peptide- secukinumab acid conjugates, the ankylosing spondylitis condition is alleviated.

[0301] The peptide can also be a peptide of SEQ ID NO: 24. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 36

Peptide-Sirukumab Conjugates

[0302] This example describes conjugation of a peptide of SEQ ID NO: 199 this disclosure to sirukumab. Sirukumab is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to sirukumab. Alternatively the peptide- active agent of this Example can be expressed as a fusion protein.

[0303] The peptide- sirukumab conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to kidneys. The subject is a human or animal and has rheumatoid arthritis, immune diseases of the kidneys. Upon administration and homing of peptide- sirukumab conjugates, the rheumatoid arthritis, immune diseases of the kidneys condition is alleviated.

[0304] The peptide can also be a peptide of SEQ ID NO: 36. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 37

Pep tide- Anifrolumab Conjugates

[0305] This example describes conjugation of a peptide of SEQ ID NO: 27 this disclosure to anifrolumab. Anifrolumab is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to

anifrolumab. Alternatively the peptide- active agent of this Example can be expressed as a fusion protein.

[0306] The peptide-anifrolumab conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to kidneys. The subject is a human or animal and has lupus nephritis. Upon administration and homing of peptide-anifrolumab conjugates, the lupus nephritis condition is alleviated.

[0307] The peptide can also be a peptide of SEQ ID NO: 185. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 38

Peptide-Denosumab Conjugates

[0308] This example describes conjugation of a peptide of SEQ ID NO: 108 this disclosure to denosumab. Denosumab is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to

denosumab. Alternatively the peptide- active agent of this Example can be expressed as a fusion protein.

[0309] The peptide-denosumab conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has osteoporosis. Upon administration and homing of peptide-denosumab conjugates, the osteoporosis condition is alleviated.

[0310] The peptide can also be a peptide of SEQ ID NO: 25. The peptide can be any peptide with the sequence selected SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 39

Peptide-Rituximab Conjugates

[0311] This example describes conjugation of a peptide of SEQ ID NO: 108 this disclosure to rituximab. Rituximab is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to rituximab. Alternatively the peptide- active agent of this Example can be expressed as a fusion protein.

[0312] The peptide-rituximab conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage or kidneys. The subject is a human or animal and has rheumatoid arthritis, kidney transplant. Upon administration and homing of peptide-rituximab conjugates, the rheumatoid arthritis, kidney transplant condition is alleviated.

[0313] The peptide can also be a peptide of SEQ ID NO: 26. The peptide can be any peptide with the sequence selected SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 40

Peptide- Omalizumab Conjugates

[0314] This example describes conjugation of a peptide of SEQ ID NO: 108 this disclosure to omalizumab. Omalizumab is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to

omalizumab. Alternatively the peptide- active agent of this Example can be expressed as a fusion protein.

[0315] The peptide-omalizumab conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to kidneys. The subject is a human or animal and has kidney inflammation. Upon administration and homing of peptide-omalizumab conjugates, the kidney inflammation condition is alleviated.

[0316] The peptide can also be a peptide of SEQ ID NO: 107. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 41

Pep tide- Abatacept Conjugates

[0317] This example describes conjugation of a peptide of SEQ ID NO: 199 this disclosure to abatacept. Abatacept is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to abatacept. Alternatively the peptide- active agent of this Example can be expressed as a fusion protein.

[0318] The peptide- abatacept conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to kidneys. The subject is a human or animal and has rheumatoid arthritis, lupus nephritis, organ transplant, focal segmental glomerulosclerosis. Upon administration and homing of peptide- abatacept conjugates, the rheumatoid arthritis, lupus nephritis, organ transplant, focal segmental glomerulosclerosis condition is alleviated.

[0319] The peptide can also be a peptide of SEQ ID NO: 111. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 42

Peptide- Oxycodone Conjugates

[0320] This example describes conjugation of a peptide of SEQ ID NO: 187 this disclosure to oxycodone. Oxycodone is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013).

[0321] The peptide-oxycodone conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has cartilage or kidney-related pain. Upon administration and homing of peptide-oxycodone conjugates, the cartilage-related pain condition is alleviated.

[0322] The peptide can also be a peptide of SEQ ID NO: 24. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 43

Peptide Capsaicin Conjugates

[0323] This example describes conjugation of a peptide of SEQ ID NO: 108 this disclosure to capsaicin. Capsaicin is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013).

[0324] The peptide-capsaicin conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has cartilage-related pain. Upon administration and homing of peptide-capsaicin conjugates, the cartilage or kidney-related pain condition is alleviated.

[0325] The peptide can also be a peptide of SEQ ID NO: 36. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22). EXAMPLE 44

Peptide-GSK2193874 Conjugates

[0326] This example describes conjugation of a peptide of SEQ ID NO: 106 this disclosure to GSK2193874. GSK2193874 is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013).

[0327] The peptide-GSK2193874 conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to

GSK2193874. The subject is a human or animal and has cartilage-related pain. Upon

administration and homing of peptide-GSK2193874 conjugates, the cartilage-related pain condition is alleviated.

[0328] The peptide can also be a peptide of SEQ ID NO: 185. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 45

Peptide BIIB023 Conjugates

[0329] This example describes conjugation of a peptide of SEQ ID NO: 199 this disclosure to BIIB023. BIIB023 is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to BIIB023. Alternatively the peptide- active agent of this Example can be expressed as a fusion protein.

[0330] The peptide-BIIB023 conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has lupus nephritis or rheumatoid arthritis. Upon administration and homing of peptide-BIIB023 conjugates, the lupus nephritis or rheumatoid arthritis condition is alleviated.

[0331] The peptide can also be a peptide of SEQ ID NO: 25. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22). EXAMPLE 46

Pep tide- Anakinra Conjugates

[0332] This example describes conjugation or fusion of a peptide of SEQ ID NO: 187 or SEQ ID NO: 550-564 of this disclosure to anakinra. A linker is optionally used to conjugate the peptide to anakinra. Anakinra is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to anakinra. Alternatively the peptide- active agent of this Example can be expressed as a fusion protein.

[0333] The peptide- anakinra conjugates or fusions are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has lupus nephritis or rheumatoid arthritis. Upon administration and homing of peptide- anakinra conjugates or fusions, the lupus nephritis or rheumatoid arthritis condition is alleviated.

[0334] . The peptide can also be a peptide of SEQ ID NO: 26. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 47

Peptide-IGF-1 Conjugates

[0335] This example describes conjugation of a peptide of SEQ ID NO: 108 this disclosure to IGF-1. IGF-1 is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to IGF-1. Alternatively the peptide- active agent (where the active agent is the biologic of this Example) can be expressed as a fusion protein.

[0336] The peptide-IGF- 1 conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has renal cancer or arthritis. Upon administration and homing of peptide-IGF- 1 conjugates, the renal cancer or arthritis condition is alleviated.

[0337] The peptide can also be a peptide of SEQ ID NO: 107. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22). EXAMPLE 48

Peptide-Romosozumab Conjugates

[0338] This example describes conjugation of a peptide of SEQ ID NO: 106 this disclosure to Romosozumab. Romosozumab is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to

romosozumab. Alternatively the peptide- active agent of this Example can be expressed as a fusion protein.

[0339] The peptide-romosozumab conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has osteoporosis. Upon administration and homing of peptide-romosozumab conjugates, the osteoporosis condition is alleviated.

[0340] The peptide can also be a peptide of SEQ ID NO: 111. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 49

Peptide-ZVAD-fmk Conjugates

[0341] This example describes conjugation of a peptide of SEQ ID NO: 187 this disclosure to ZVAD-fmk. ZVAD-fmk is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to ZVAD- fmk. The peptide- ZVAD-fmk conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has cartilage grafting, arthritis, surgical intervention, surgery for cartilage repair. Upon administration and homing of peptide- ZVAD-fmk conjugates, the cartilage grafting, arthritis, surgical intervention, surgery for cartilage repair condition is alleviated.

[0342] The peptide can also be a peptide of SEQ ID NO: 24. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22). EXAMPLE 50

Peptide- S -methylisothiourea Conj ugates

[0343] This example describes conjugation of a peptide of SEQ ID NO: 108 this disclosure to S- methylisothiourea. S-methylisothiourea is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013).

[0344] The peptide-S-methylisothiourea conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has arthritis surgery, kidney iron overload, renal ischemia reperfusion injury, or acute kidney injury. Upon administration and homing of peptide- S-methylisothiourea conjugates, the arthritis surgery, kidney iron overload, renal ischemia reperfusion injury, or acute kidney injury condition is alleviated.

[0345] The peptide can also be a peptide of SEQ ID NO: 33. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 51

Peptide-P188 Conjugates

[0346] This example describes conjugation of a peptide of SEQ ID NO: 106 this disclosure to P188. P188 is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013).

[0347] The peptide-P188 conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has arthritis surgery. Upon administration and homing of peptide-P188 conjugates, the arthritis surgery condition is alleviated.

[0348] The peptide can also be a peptide of SEQ ID NO: 185. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22). EXAMPLE 52

Pep tide- Alendronate Conjugates

[0349] This example describes conjugation of a peptide of SEQ ID NO: 187 this disclosure to alendronate. Alendronate is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013).

[0350] The peptide-alendronate conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has bone erosion. Upon administration and homing of peptide-alendronate conjugates, the bone erosion condition is alleviated.

[0351] The peptide can also be a peptide of SEQ ID NO: 22. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 53

Peptide-MIP-3a Conjugates

[0352] This example describes conjugation of a peptide of SEQ ID NO: 108 this disclosure to MIP-3a. MIP-3a is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to MIP-3a. Alternatively the peptide- active agent of this Example can be expressed as a fusion protein

[0353] The peptide-MIP-3a conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has Joint injury, repair and regeneration of cartilage and bone. Upon administration and homing of peptide-MIP-3a conjugates, the Joint injury, repair and regeneration of cartilage and bone condition is alleviated.

[0354] The peptide can also be a peptide of SEQ ID NO: 26. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22). EXAMPLE 54

Peptide-BMP-2 Conjugates

[0355] This example describes conjugation of a peptide of SEQ ID NO: 106 this disclosure to BMP-2. BMP-2 is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to BMP-2. Alternatively the peptide- active agent of this Example can be expressed as a fusion protein.

[0356] The peptide-BMP-2conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has Joint repair. Upon administration and homing of peptide- BMP-2conjugates, the Joint repair condition is alleviated.

[0357] The peptide can also be a peptide of SEQ ID NO: 107. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 55

Peptide-Icariin Conjugates

[0358] This example describes conjugation of a peptide of SEQ ID NO: 187 this disclosure to icariin. Icariin is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013).

[0359] The peptide-icariin conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has Joint repair. Upon administration and homing of peptide- icariin conjugates, the Joint repair condition is alleviated.

[0360] The peptide can also be a peptide of SEQ ID NO: 108. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or a stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 56

Peptide- Cap topril Conjugates

[0361] This example describes conjugation of a peptide of SEQ ID NO: 27 this disclosure to captopril. Captopril is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013).

[0362] The peptide-captopril conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to kidneys. The subject is a human or animal and has diabetic nephropathy. Upon administration and homing of peptide-captopril conjugates, the diabetic nephropathy condition is alleviated.

[0363] The peptide can also be a peptide of SEQ ID NO: 24. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 57

Peptide- Tofacitinib Conjugates

[0364] This example describes conjugation of a peptide of SEQ ID NO: 108 this disclosure to tofacitinib. Tofacitinib is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). From one to eight peptides are linked to tofacitinib.

[0365] The peptide-tofacitinib conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage. The subject is a human or animal and has rheumatoid arthritis and kidney transplant, ankyloses spondylitis. Upon administration and homing of peptide-tofacitinib conjugates, the rheumatoid arthritis and kidney transplant, ankyloses spondylitis condition is alleviated.

[0366] The peptide can also be a peptide of SEQ ID NO: 36. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 58

Peptide-Dimethyl fumarate Conjugates

[0367] This example describes conjugation of a peptide of SEQ ID NO: 108 this disclosure to dimethyl fumarate. Dimethyl fumarate is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013). Alternatively, peptide-dimethyl fumarate conjugates can be synthesized by Michael addition of a thiol (on the peptide of linker) to dimethyl fumarate as described by Schmidt et al. (Bioorg Med Chem. 2007 Jan l;15(l):333-42. Epub 2006 Sep 29. ).

[0368] The peptide-dimethyl fumarate conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to kidneys. The subject is a human or animal and has Kidney fibrosis, psoriatic arthritis, rheumatoid arthritis. Upon administration and homing of peptide-dimethyl fumarate conjugates, the Kidney fibrosis, psoriatic arthritis, rheumatoid arthritis condition is alleviated.

[0369] The peptide can also be a peptide of SEQ ID NO: 187. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 59

Intra-articular Administration of Peptides and Peptide Conjugates

[0370] This example illustrates intra-articular administration of peptides or peptide conjugates of this disclosure. A peptide of this disclosure is expressed recombinantly or chemically

synthesized. In some cases, the peptide is subsequently conjugated to a detectable agent or an active agent. The peptide or peptide conjugate is administered to a subject in need thereof via intra-articular administration. The cartilage is penetrated by the peptide or peptide conjugate due to the small size of the peptide or peptide conjugate, and due to binding of cartilage components by the peptide or peptide conjugate. The peptide or peptide conjugate is bound to cartilage and the residence time in the cartilage is longer due to this binding. Optionally, the injected material is aggregated, is crystallized, or complexes are formed, further extending the depot effect and contributing to longer residence time.

[0371] The peptide can be a peptide of SEQ ID NO: 108. The peptide can also be a peptide of SEQ ID NO: 24. The peptide can be any peptide with the sequence selected SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 60

Treatment of Osteoarthritis

[0372] This example describes a method for treating osteoarthritis using peptides of the present disclosure. This method is used as a treatment for acute and/or chronic symptoms associated with osteoarthritis. A peptide of the present disclosure is expressed recombinantly or chemically synthesized and then is used directly or conjugated to an anti- inflammatory compound, such as aspirin, desciclesonide, or secukinumab. The resulting peptide or peptide-drug conjugate is administered in a pharmaceutical composition subcutaneously, intravenously, or orally, or is injected directly into a joint of a patient and targeted to cartilage. The formulation can be modified physically or chemically to increase the time of exposure in the cartilage. One or more anti- inflammatory peptide conjugates are administered to a human or animal.

[0373] The peptide can be a peptide of SEQ ID NO: 106. The peptide can also be a peptide of SEQ ID NO: 33. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 61

Treatment of Cartilage Degradation

[0374] This example describes a method for treating and/or preventing cartilage degradation using a peptide of the present disclosure. This method is used as a treatment for acute and/or chronic symptoms associated with cartilage degradation. Progressive degradation or thinning of the cartilage is difficult to treat in part because molecules such as small molecule drugs and antibodies typically do not reach the avascular cartilage. A peptide of the present disclosure is used for its homing and/or native activity, or is mutated to generate activity such as MMP protease inhibition. It is expressed recombinantly or chemically synthesized and then is used directly or conjugated to an extracellular matrix targeting active agent, such as an inhibitor of MMP activity or an anti-apoptosis agent (e.g., osteoprotegrin, romosozumab, P188, ZVAD-fmk, quercetin, dasatinib, dimethyl fumarate, bortezomib, carilzomib, or navitoclax). The resulting peptide or peptide-drug conjugate is administered in a pharmaceutical composition

subcutaneously, intravenously, or orally, or is injected directly into a joint of a patient and targeted to extracellular matrix. One or more extracellular matrix targeting conjugates are administered to a human or animal.

[0375] The peptide can be a peptide of SEQ ID NO: 187. The peptide can also be a peptide of SEQ ID NO: 27. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22). EXAMPLE 62

Treatment of a Cartilage Injury

[0376] This example describes a method for treating a cartilage injury using a peptide of the present disclosure. A peptide of the present disclosure is expressed recombinantly or chemically synthesized and then is used directly or conjugated to a therapeutic compound, such as those described herein, including, but not limited to BMP-2, BMP-7, BMP-9, BMP- 13, PDGF, PTH, PTHrP, IL-8, MIP-3oc. The resulting peptide or peptide-drug conjugate is administered in a pharmaceutical composition to a patient and targeted to cartilage. One or more therapeutic compound-peptide conjugates are administered to a human or animal.

[0377] The peptide can be a peptide of SEQ ID NO: 108. The peptide can also be a peptide of SEQ ID NO: 185. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 63

Treatment of Rheumatoid Arthritis

[0378] This example describes a method for treating rheumatoid arthritis. This method is used as a treatment for acute and/or chronic symptoms associated with rheumatoid arthritis. A peptide of the present disclosure is expressed recombinantly or chemically synthesized and then is used directly, or is conjugated to an anti- inflammatory compound, such as adalimumab, certohzumab, golimumab, thalidomide, lenalidomide, pomalidomide, pentocifylline, bupropion. When the peptide is used directly, the peptide can, for example, bind or inhibit ion channels such as Kv 1. 3. The resulting peptide or peptide-drug conjugate is administered in a pharmaceutical composition to a patient and is targeted to cartilage. One or more anti- inflammatory compound- peptide conjugates are administered to a human or animal subcutaneously, intravenously, or orally, or is injected directly into a joint

[0379] The peptide can be a peptide of SEQ ID NO: 106. The peptide can also be a peptide of SEQ ID NO: 25. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22). EXAMPLE 64

Treatment of Gout

[0380] This example describes a method for treating gout using peptides of the present disclosure. This method is used as a treatment for acute and/or chronic symptoms associated with gout. A peptide of the present disclosure is expressed and administered in a pharmaceutical composition to a patient as a therapeutic for gout. A peptide of the disclosure is recombinantly or chemically synthesized and then is used directly or conjugated to pegloticase to treat a cartilage disorder. A peptide of the disclosure is recombinantly or chemically synthesized and then is used directly or conjugated to probenecid to treat a kidney disorder. The peptide is administered in a pharmaceutical composition to a patient and the peptide is targeted to the cartilage or kidney affected by gout. One or more peptides are administered to a human or animal subcutaneously, intravenously, or orally, or is injected directly into a joint.

[0381] The peptide can be a peptide of SEQ ID NO: 187. The peptide can also be a peptide of SEQ ID NO: 24. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 65

Treatment or Management of Pain

[0382] This example describes a method for treating or managing pain associated with a cartilage injury or disorder. This method is used as a treatment for acute and/or chronic symptoms associated with a cartilage injury or disorder. A peptide of the disclosure is expressed and administered in a pharmaceutical composition to a patient as a therapeutic for pain as a result of injury or other cartilage or joint condition as described herein. The peptide of the present disclosure inhibits ion channels, such as Nav 1.7. The peptide is expressed recombinantly or chemically synthesized, wherein the peptide selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Alternatively, the peptides of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564 are mutated to maintain the cartilage homing function, but to add or increase ion channel inhibition, such as to Nav 1. 7. Following expression or synthesis, the peptide is used directly or conjugated to a narcotic (e.g., oxycodone), a nonnarcotic analgesic, a natural counter-irritant (capsaicin), or a pain receptor channel inhibitor (such as the TRPV4 inhibitor GSK2193874). Following administration of the peptide, the peptide targets to the cartilage affected by pain. One or more peptides are administered to a human or animal subcutaneously, intravenously, or orally, or is injected directly into a joint. [0383] The peptide can be a peptide of SEQ ID NO: 108. The peptide can also be a peptide of SEQ ID NO: 107. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 66

Treatment or Management of Pain with Peptides Only

[0384] This example describes a method for treating or managing pain associated with a cartilage injury or disorder. This method is used as a treatment for acute and/or chronic symptoms associated with a cartilage injury or disorder. A peptide of the disclosure is expressed and administered in a pharmaceutical composition to a patient as a therapeutic for pain as a result of injury or other cartilage or joint condition as described herein. The peptide of the present disclosure inhibits ion channels, such as Nav 1.7. The peptide is expressed recombinantly or chemically synthesized, wherein the peptide selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Alternatively, the peptides of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564 are mutated to maintain the cartilage homing function, but to add or increase ion channel inhibition, such as to Nav 1.7. Following expression or synthesis, the peptide is used directly. Following administration of the peptide, the peptide targets to the cartilage affected by pain. One or more peptides are administered to a human or animal subcutaneously, intravenously, or orally, or is injected directly into a joint.

[0385] The peptide can be a peptide of SEQ ID NO: 108. The peptide can also be a peptide of SEQ ID NO: 107. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564.

EXAMPLE 67

Treatment of Chondrosarcoma

[0386] This example illustrates treatment of chondrosarcoma using peptides of the present disclosure. A peptide of the present disclosure is recombinantly expressed or chemically synthesized and are used directly, after radiolabeling, or after conjugation to a fluorophore or therapeutic compound, such as dasatinib. The peptide or peptide conjugate is administered in a pharmaceutical composition to a subject as a therapeutic for chondrosarcoma. One or more peptides or peptide conjugates of the present disclosure are administered to a subject. A subject can be a human or an animal. The pharmaceutical composition is administered subcutaneously, intravenously, orally, or injected directly into a joint. The peptides or peptide conjugates target cartilage affected by chondrosarcoma.

[0387] The peptide can be a peptide of SEQ ID NO: 106. The peptide can also be a peptide of SEQ ID NO: 108. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 68

Treatment of Chordoma

[0388] This example illustrates treatment of chordoma using peptides of the present disclosure. A peptide of the present disclosure is recombinantly expressed or chemically synthesized and are used directly, after radiolabeling, or after conjugation to a fluorophore or therapeutic compound, such as dasatinib. The peptide or peptide conjugate is administered in a pharmaceutical composition to a subject as a therapeutic for chordoma. One or more peptides or peptide conjugates of the present disclosure are administered to a subject. A subject can be a human or an animal. The pharmaceutical composition is administered subcutaneously, intravenously, orally, or injected directly into a joint. The peptides or peptide conjugates target cartilage affected by chordoma.

[0389] The peptide can be a peptide of SEQ ID NO: 187. The peptide can also be a peptide of SEQ ID NO: 24. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 69

Treatment for Rapid Pain Relief

[0390] This example illustrates rapid pain relief in patients treated for rheumatoid arthritis or osteoarthritis with the peptides or peptide conjugates of this disclosure. A peptide of this disclosure is expressed recombinantly or chemically synthesized, and then the N-terminus of the peptide is conjugated to an active agent via an NHS ester to produce a peptide- active agent conjugate. In some aspects the active agent such as a kidney therapeutic from TABLE 4, TABLE 5, or TABLE 6. In some cases, the peptide alone is administered to the subject.

[0391] The peptide or peptide- active agent conjugate is administered to a subject in need thereof. The subject is a human or non-human animal. The subject in need thereof has rheumatoid arthritis or osteoarthritis. The peptide or peptide conjugate is delivered via intravenous administration. Upon administration, the peptide or peptide conjugate rapidly homes to cartilage. Rapid pain relief within five minutes to an hour is experienced by the subject, and pain relieve can last as long as over 3 hours.

[0392] The peptide can be a peptide of SEQ ID NO: 108. The peptide can also be a peptide of SEQ ID NO: 33. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 70

Treatment for Lupus Nephritis

[0393] This example illustrates treatment of lupus nephritis using peptides or peptide conjugates of this disclosure. A peptide of the present disclosure is recombinantly expressed or chemically synthesized and are used directly, after radiolabeling, or after conjugation to a fluorophore or therapeutic compound, such as abatacept or BIIB023.

[0394] The peptide or peptide conjugate is administered in a pharmaceutical composition to a subject as a therapeutic for lupus nephritis. The peptide is selected from any one of the peptides of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. One or more peptides or peptide conjugates of the present disclosure are administered to a subject. A subject can be a human or an animal. The pharmaceutical composition is administered subcutaneously, intravenously, orally, or injected directly. The peptides or peptide conjugates target kidney affected by lupus nephritis.

[0395] The peptide can be a peptide of SEQ ID NO: 27. The peptide can also be a peptide of SEQ ID NO: 24. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 71

Treatment for Acute Kidney Injury (AKI)

[0396] This example illustrates treatment of acute kidney injury (AKI) using peptides or peptide conjugates of this disclosure. A peptide of the present disclosure is recombinantly expressed or chemically synthesized and are used directly, after radiolabeling, or after conjugation to a fluorophore or therapeutic compound, such as such as a kidney therapeutic from TABLE 4, TABLE 5, or TABLE 6.

[0397] The peptide or peptide conjugate is administered in a pharmaceutical composition to a subject as a therapeutic for acute kidney injury (AKI). The peptide is selected from any one of the peptides of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. One or more peptides or peptide conjugates of the present disclosure are administered to a subject. A subject can be a human or an animal. The pharmaceutical composition is administered subcutaneously, intravenously, orally, or injected directly into a joint. The peptides or peptide conjugates target cartilage affected by acute kidney injury (AKI).

[0398] The peptide can be a peptide of SEQ ID NO: 108. The peptide can also be a peptide of SEQ ID NO: 36. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 72

Treatment for Chronic Kidney Disease (CKD)

[0399] This example illustrates treatment of chronic kidney disease (CKD) using peptides or peptide conjugates of this disclosure. A peptide of the present disclosure is recombinantly expressed or chemically synthesized and are used directly, after radio labeling, or after conjugation to a fluorophore or therapeutic compound, such as a kidney therapeutic from

TABLE 4, TABLE 5, or TABLE 6.

[0400] The peptide or peptide conjugate is administered in a pharmaceutical composition to a subject as a therapeutic for chronic kidney disease (CKD). The peptide is selected from any one of the peptides of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. One or more peptides or peptide conjugates of the present disclosure are administered to a subject. A subject can be a human or an animal. The pharmaceutical composition is administered subcutaneously, intravenously, orally, or injected directly into a joint. The peptides or peptide conjugates target cartilage affected by chronic kidney disease (CKD).

[0401] The peptide can be a peptide of SEQ ID NO: 199. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22). EXAMPLE 73

Treatment for Hypertensive Kidney Damage

[0402] This example illustrates treatment of hypertensive kidney damage using peptides or peptide conjugates of this disclosure. A peptide of the present disclosure is recombinantly expressed or chemically synthesized and are used directly, after radio labeling, or after conjugation to a fluorophore or therapeutic compound, such as such as a kidney therapeutic from TABLE 4, TABLE 5, or TABLE 6.

[0403] The peptide or peptide conjugate is administered in a pharmaceutical composition to a subject as a therapeutic for hypertensive kidney damage. The peptide is selected from any one of the peptides of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. One or more peptides or peptide conjugates of the present disclosure are administered to a subject. A subject can be a human or an animal. The pharmaceutical composition is administered subcutaneously, intravenously, orally, or injected directly into a joint. The peptides or peptide conjugates target cartilage affected by hypertensive kidney damage.

[0404] The peptide can be a peptide of SEQ ID NO: 27. The peptide can also be a peptide of SEQ ID NO: 185. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 74

Treatment for Diabetic Nephropathy

[0405] This example illustrates treatment of diabetic nephropathy using peptides or peptide conjugates of this disclosure. A peptide of the present disclosure is recombinantly expressed or chemically synthesized and are used directly, after radio labeling, or after conjugation to a fluorophore or therapeutic compound, such as such as a kidney therapeutic from TABLE 4, TABLE 5, or TABLE 6.

[0406] The peptide or peptide conjugate is administered in a pharmaceutical composition to a subject as a therapeutic for diabetic nephropathy. The peptide is selected from any one of the peptides of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. One or more peptides or peptide conjugates of the present disclosure are administered to a subject. A subject can be a human or an animal. The pharmaceutical composition is administered subcutaneously, intravenously, orally, or injected directly into a joint. The peptides or peptide conjugates target cartilage affected by diabetic nephropathy. [0407] The peptide can be a peptide of SEQ ID NO: 108. The peptide can also be a peptide of SEQ ID NO: 22. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 75

Treatment for Renal Fibrosis

[0408] This example illustrates treatment of renal fibrosis using peptides or peptide conjugates of this disclosure. A peptide of the present disclosure is recombinantly expressed or chemically synthesized and are used directly, after radiolabeling, or after conjugation to a fluorophore or therapeutic compound, such as such as a kidney therapeutic from TABLE 4, TABLE 5, or

TABLE 6.

[0409] The peptide or peptide conjugate is administered in a pharmaceutical composition to a subject as a therapeutic for renal fibrosis. The peptide is selected from any one of the peptides of SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. One or more peptides or peptide conjugates of the present disclosure are administered to a subject. A subject can be a human or an animal. The pharmaceutical composition is administered subcutaneously, intravenously, orally, or injected directly into a joint. The peptides or peptide conjugates target cartilage affected by renal fibrosis.

[0410] The peptide can be a peptide of SEQ ID NO: 199. The peptide can also be a peptide of SEQ ID NO: 26. The peptide can be any peptide with the sequence selected from SEQ ID NO: 24 - SEQ ID NO: 274 or SEQ ID NO: 314 - SEQ ID NO: 564. Such peptide-drug conjugates can be made using either a cleavable or stable linker as described herein (e.g., EXAMPLES 21 and 22).

EXAMPLE 76

Peptide Variants Based on Multiple Sequence Alignment

[0411] This example illustrates using multiple sequence alignment to design peptide variants with increased stability and decreased immunogenicity. An alignment was generated using R language and an "msa" software package, which codes for R language specific for multiple alignments (Bodenhofer, U et al. Bioinformatics, 31 (24): 3997-3999 (2015)). FIG. 11 illustrates a multiple sequence alignment of SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO: 321, SEQ ID NO: 333, SEQ ID NO: 337, SEQ ID NO: 338, SEQ ID NO: 340, SEQ ID NO: 398, SEQ ID NO: 474, SEQ ID NO: 483, SEQ ID NO: 486, and SEQ ID NO: 543 - SEQ ID NO: 549. The alignment identified permissive or preferred amino acids at a given location, and provided a guide for discovery of novel peptide variants that could be generated and that could retain essential properties such as structure, function, peptide folding, biodistribution, or stability. SEQ ID NO: 21 and SEQ ID NO: 295 are consensus sequences based on the above multiple sequence alignment. SEQ ID NO: 21 is the same sequence as SEQ ID NO: 295 but with an N-terminal "GS." Furthermore, based on the ability to substitute K residues to R residues, the multiple sequence alignment identified peptides of the family of sequences of SEQ ID NO: 22 and SEQ ID NO: 296 as potential peptide variants that could be generated and that could retain essential properties such as structure, function, peptide folding, biodistribution, or stability. Additionally, the multiple sequence alignment identified SEQ ID NO: 312 as a conserved region within the sequences of the alignment, which may, at least in part, be important for maintaining the essential properties such as structure, function, peptide folding, biodistribution, binding, accumulation, retention, or stability.

EXAMPLE 77

Peptide Immunogenicity

[0412] This example illustrates the testing of the immunogenicity of a peptide. NetMHC II version 2.3 prediction software was used to identify immunogenic peptides based on a neural network alignment algorithm that predicts peptide binding to MHC Class II molecules.

The NetMHC II prediction software was utilized to determine the putative peptide binding capability to DR, DQ, and DP MHC II alleles and the strength of the interaction between peptide and MHC II molecules. TABLE 7 shows the resulting immunogenicity score of select peptides. The numbers of strong versus weak peptides were tallied into each major MHC allele group (DR, DQ, and DP). Additionally, the numbers of 'unique strong' and 'unique weak core' peptides were also tallied. These data were used to predict which peptides are less likely to induce an immunogenic response in patients. For example, the stronger a peptide binds to an allele, the more likely it is to be presented in a MHC/peptide combination on an antigen presenting cell, thus triggering an immune response, and a peptide that is predicted to bind to fewer alleles is more likely to have weaker binding to given alleles and should be less immunogenic.

TABLE 7- Immunogenicity Scores of Peptides

SEQ ID NO: Strong Binding Unique Weak Binding Alleles Unique Weak

Alleles Strong Core (DR + DQ + DP) Core Peptides (DR + DQ + Peptides

DP)

108 1 + 0 + 0 1 + 0 + 0 7 + 1 + 0 7 + 2 + 0

260 0 + 0 + 0 0 + 0 + 0 4 + 1 + 3 6 + 1 + 1

261 0 + 0 + 0 0 + 0 + 0 4 + 1 + 3 6 + 2 + 1

262 1 + 0 + 0 2 + 0 + 0 5 + 1 + 3 7 + 1 + 1

EXAMPLE 78

Peptide Variants

[0413] This example illustrates the design of variant peptide sequences with increased stability, decreased regions of immunogenicity, and the substitution of a tyrosine for spectrophoto metric reporting as compared to a parent peptide sequence. Potential mutations to the parent peptide sequence, SEQ ID NO: 108, that may result in a peptide with increased stability, decreased immunogenicity, or increased absorbance at 270-280 nm (such as the substitution to a tyrosine or tryptophan residue for spectrophotometric reporting) were identified based on information from multiple sequence alignment from EXAMPLE 76 and immunogenicity testing from EXAMPLE 77.

[0414] In SEQ ID NO: 108, residue N7 is at risk for deamidation. Based on the multiple sequence alignment of SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO: 321, SEQ ID NO: 333, SEQ ID NO: 337, SEQ ID NO: 338, SEQ ID NO: 340, SEQ ID NO: 398, SEQ ID NO: 483, SEQ ID NO: 486, and SEQ ID NO: 543 - SEQ ID NO: 549, the candidate residue mutations to best reduce this risk were N7S and N7G. N7S was determined to be more likely to result in a peptide with desirable properties such as folding and stability as shown by matches in the alignment and conservationist presence in a peptide with high stability (SEQ ID NO: 474).

[0415] Residue D18 is at risk for cleavage. Based on the multiple sequence alignment, the candidate residue mutations to best reduce cleavage at D18 are D18E and D18Q. D18E is the preferred choice based on retaining charge.

[0416] Residue M25 is at risk for oxidation. Based on the multiple sequence alignment, the candidate residue mutations to best reduce oxidation were M25T and M25A. Based on the immunogenicity score of peptides with each mutation, it was determined that M25T is the better mutation, as it eliminates a significant source of immunogenicity as compared to SEQ ID NO: 108 as well as the variant with M25A, which did not eliminate the predicted immunogenicity of the parent peptide of SEQ ID NO: 108.

[0417] Residue N32 is at risk for deamidation, at least in part due to the neighboring residue S33. However, N32 is conserved across Kvl. 3 binding cystine-dense peptides in the alignment of EXAMPLE 76, and implicated in receptor binding (Peigneur, S., Biochemistry, 55(32): 2927-35 (2016)). For certain applications, peptides are designed to maintain this binding interaction, and for other applications, peptides are designed to remove this binding interaction. To maintain functionality, one candidate residue mutation based on the multiple sequence alignment is S33R, which would impact deamidation. However, it resulted in a predicted increased immunogenicity score. Another candidate residue mutation is S33G, but this may result in higher deamidation rates. If N32 is mutated, the best candidate residue mutation based the multiple sequence alignment in combination with the immunogenicity score was N32Q despite it having a slight increase in immunogenicity. Other options are N32A, N32S, or N32T. Alternatively, to remove functionality, candidate mutations based on the multiple sequence alignment are N32A and N32L, which are the preferred choices.

[0418] For the substitution to a tyrosine for spectrophotometric reporting, the best candidate locations were T38Y (which had the strongest precedence in the multiple sequence alignment and is found in several of the stable peptides (e.g., SEQ ID NO: 474, SEQ ID NO: 544, and SEQ ID NO: 545)), L17Y, and H36Y. However, T38Y may slightly increase immunogenicity with respect to the DR allele. Another option for spectrophometric absorbance is to substitute Trp for the Leu at position 17.

[0419] Based on the above analysis, the following short list of potential mutations for SEQ ID NO: 108 were compiled: N7S; D18E; M25T; N32Q, N32A, N32S, N32T, N32L, S33G, and S33R (variants both to retain function and to remove function of binding ion channel); and L17Y, H36Y, and T38Y.

TABLE 8 provides some exemplary sequences using various combinations of these mutations.

TABLE 8 - Exemplary Sequence Variants of SEQ ID NO: 108

SEQ ID NO: Mutations

556 L15Y, D16E, M23T, N30Q

557 D16E, M23T, N30Q, H34Y

558 N5S, D16E, M23T, N30Q, T36Y

559 N5S, L15Y, D16E, M23T, N30Q

560 N5S, D16E, M23T, N30Q, H34Y

561 D16E, M23T, N32A, T36Y

562 D16E, M23T, N32S, T36Y

563 D16E, M23T, N32T, T36Y

564 D16E, M23T, T36Y

EXAMPLE 79

Peptide-Budesonide Conjugate

[0420] This example describes conjugation of a peptide of any one of SEQ ID NO: 260 - SEQ ID NO: 274 or SEQ ID NO: 550 - SEQ ID NO: 564 to budesonide. Budesonide is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013) or by any of the methods described in EXAMPLES 25-28.

[0421] The peptide-budesonide conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage and/or kidneys. The subject is a human or animal and has inflammation in the cartilage or kidney tissues. Upon administration and homing of peptide-budesonide conjugates, the inflammation in the cartilage and/or kidney tissues is alleviated.

EXAMPLE 80

Peptide-Dexamethasone Conjugate

[0422] This example describes conjugation of a peptide of any one of SEQ ID NO: 260 - SEQ ID NO: 274 or SEQ ID NO: 550 - SEQ ID NO: 564 to dexamethasone. Dexamethasone is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013) or by any of the methods described in EXAMPLES 25-28.

[0423] The peptide-dexamethasone conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage and/or kidneys. The subject is a human or animal and has inflammation in the cartilage or kidney tissues. Upon administration and homing of peptide-dexamethasone conjugates, the inflammation in the cartilage and/or kidney tissues is alleviated. EXAMPLE 81

Peptide- Triamcinalone Acetonide Conjugate

[0424] This example describes conjugation of a peptide of any one of SEQ ID NO: 260 - SEQ ID NO: 274 or SEQ ID NO: 550 - SEQ ID NO: 564 to triamicinalone acetonide. Triamicinalone acetonide is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013 or by any of the methods described in EXAMPLES 25-28.

[0425] The peptide-triamicinalone acetonide conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage and/or kidneys. The subject is a human or animal and has inflammation in the cartilage or kidney tissues. Upon administration and homing of peptide-triamicinalone acetonide conjugates, the inflammation in the cartilage and/or kidney tissues is alleviated.

EXAMPLE 82

Peptide-Desciclesonide Acetonide Conjugate

[0426] This example describes conjugation of a peptide of any one of SEQ ID NO: 260 - SEQ ID NO: 274 or SEQ ID NO: 550 - SEQ ID NO: 564 to desciclesonide acetonide. Desciclesonide acetonide is readily conjugated to any peptide disclosed herein via standard chemistries such as those described in, but not limited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3 ^rd edition, 2013) or by any of the methods described in EXAMPLES 25-28.

[0427] The peptide-desciclesonide acetonide conjugates are administered to a subject in need thereof and home, target, are directed to, are retained by, accumulate in, migrate to, and/or bind to cartilage and/or kidneys. The subject is a human or animal and has inflammation in the cartilage or kidney tissues. Upon administration and homing of peptide-desciclesonide acetonide conjugates, the inflammation in the cartilage and/or kidney tissues is alleviated.

EXAMPLE 83

Method of Peptide Synthesis

[0428] This example describes the synthesis of SEQ ID NO: 106, SEQ ID NO: 108, and SEQ ID NO: 187.

[0429] A peptide of SEQ ID NO: 108 was made using Solid Phase Peptide Synthesis (SPPS). After release of the peptide from the solid phase, the peptide was purified prior to folding by oxidation in solution. The folded peptide was further purified by reversed-phase chromatography and lyophilized as a TFA salt. The final SEQ ID NO: 108 peptide product had a purity of 96.1% and a mass of 4,301.7 Da, which confirmed its identity as a peptide of SEQ ID NO: 108. [0430] A peptide of SEQ ID NO: 106 was made using Solid Phase Peptide Synthesis (SPPS). After release of the peptide from the solid phase, the peptide was folded by oxidation in solution. The folded peptide was purified by reversed-phase chromatography and lyophilized as a TFA salt. The final SEQ ID NO: 106 had a purity of 95.6% and a mass of 4,503.0 Da, which confirmed its identity as a peptide of SEQ ID NO: 106.

[0431] A peptide of SEQ ID NO: 187 was made using Solid Phase Peptide Synthesis (SPPS). After release of the peptide from the solid phase, the peptide was folded by oxidation in solution. The folded peptide was purified by reversed-phase chromatography and lyophilized as a TFA salt. The final SEQ ID NO: 187 peptide product had a purity of 95.5% and a mass of 4,154.0 Da, which confirmed its identity as a peptide of SEQ ID NO: 187.

EXAMPLE 84

Whole Body Autoradiography of Cartilage Homing Peptides

[0432] This example illustrates peptide homing to cartilage mice 5 minutes to 48 hours after administration of a radiolabeled peptide. Signal from the radiolabeled peptides was found in all types of cartilage at each time point examined. Each peptide was radiolabeled by methylating lysines at the N-terminus as described in EXAMPLE 2. As such, the peptide may contain methyl or dimethyl lysines and a methylated or dimethlyated amino terminus. A dose of 100 nmol radiolabeled peptide was administered via tail vein injection in Female Harlan athymic nude mice, weighing 20-25 g. The experiment was done in duplicate (n=2 animals per group). Each radiolabeled peptide was allowed to freely circulate within the animal for the described time period before the animals were euthanized and sectioned.

[0433] Whole body autoradiography (WBA) sagittal sectioning was performed as follows. At the end of the dosing period, mice were frozen in a hexane/dry ice bath and then embedded in a frozen block of carboxymethylcellulose. Whole animal sagittal slices were prepared that resulted in thin frozen sections for imaging. Sections were allowed to dessicate in a freezer prior to imaging. For the autoradiography imaging, tape mounted thin sections were freeze dried and radioactive samples were exposed to phosphoimager plates. These plates were developed and the signal (densitometry) from each organ was normalized to the signal found in the cardiac blood of each animal. A signal in tissue darker than the signal expected from blood in that tissue indicates accumulation in a region, tissue, structure, or cell.

[0434] FIG. 14 illustrates autoradiography image of frozen sections from a mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 108. FIG. 14A illustrates the ¹⁴ C signal in a frozen section of a mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 108. The ¹⁴ C signal identifies the radiolabeled peptide distribution in the cartilage of the mouse. FIG. 14B illustrates the ¹⁴ C signal in a different frozen section of a mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 108. The ¹⁴ C signal identifies the radiolabeled peptide distribution in the cartilage of the mouse.

[0435] TABLE 9 shows the signal of radiolabeled peptides of SEQ ID NO: 27 and SEQ ID NO: 108 in intervertebral discs (IVD) and knee joints as a percentage of the blood. Because the peptides may arrive at the joint within five minutes, a therapeutic effect from the peptide or a conjugated active agent may begin quickly. A therapeutic effect could be long lasting, due to continued presence of detected agents at 48 hours and/or due to long lasting pharmacodynamics effects.

TABLE 9 - Signal of Radiolabeled Peptides of SEQ ID NO: 27 and SEQ ID NO: 108 in

IVD and Knee Joints as a Percentage of Blood

[0436] FIG. 15 illustrates autoradiography images of frozen sections from a mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 106. FIG. 15A illustrates the ¹⁴ C signal in a frozen section of a mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 106. The ¹⁴ C signal identifies the radiolabeled peptide distribution in the cartilage of the mouse. FIG. 15B illustrates the ¹⁴ C signal in a frozen section of a different mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 106. The ¹⁴ C signal identifies the radiolabeled peptide distribution in the cartilage of the mouse.

[0437] FIG. 16 illustrates autoradiography images of frozen sections from a mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 187. FIG. 16A illustrates the ¹⁴ C signal in a frozen section of the mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 187. The ¹⁴ C signal identifies the radiolabeled peptide distribution in the cartilage of the mouse. FIG. 16B illustrates the ¹⁴ C signal in a frozen section of a different mouse, 3 hours after administration of 100 nmol of a radiolabeled peptide of SEQ ID NO: 187. The ¹⁴ C signal identifies the radiolabeled peptide distribution in the cartilage of the mouse. [0438] This data illustrates peptides of SEQ ID NO: 27, SEQ ID NO: 108, SEQ ID NO: 106 and SEQ ID NO: 187 homed to and accumulated in the cartilage of the animals. The peptide of SEQ ID NO: 108 is a K to R variant of a peptide of SEQ ID NO: 27. These data show that K to R variants of cartilage homing peptides retained their cartilage homing properties.

[0439] SEQ ID NO: 567 (GS G VPIN VRS RGS RDS LDPS RR AGMRFGRS INS RS HS TP) is a linearized version of SEQ ID NO: 108, where the knotted scaffold of the peptide was removed by mutating out the cysteine residues that form the disulfide bonds of the peptide to serine residues, but retaining the rest of the sequence. TABLE 10 shows quantification of signal as a percentage of signal in blood from a linearized radiolabeled SEQ ID NO: 567 peptide in intervertebral discs (IVD).

TABLE 10 - Signal of Radiolabled Peptides of SEQ ID NO: 567 in IVD as a Percentage of

Blood

[0440] The peptide of SEQ ID NO: 567, a linearized version of the peptide of SEQ ID NO: 108, homed to cartilage to a much lesser extent than the folded knotted peptide (SEQ ID NO: 108). The signal of the folded knotted peptide of SEQ ID NO: 108 was ~20-fold greater at 3 hours and ~50-fold greater at 24 hours (TABLE 9) as compared to the linearized peptide of SEQ ID NO: 567 (TABLE 10). These results indicate that in addition to changes in primary sequence or peptide charge, homing to cartilage can also be related to changes in conformation, or tertiary structure. Namely, in some cases, folded cystine-dense peptides can be exemplary cartilage homers in comparison to unfolded, linearized peptides of the same primary sequence (except for the mutated cysteine residues).

EXAMPLE 85

Fluorescence of Cartilage Homing Peptides

[0441] This example illustrates peptide homing to cartilage mice after administration of a peptide fluorophore conjugate. A peptide of SEQ ID NO: 108 was chemically conjugated to one molecule of Cyanine 5.5, and then imaged using the methods of EXAMPLE 13.

[0442] FIG. 10 shows white light images and corresponding whole body fluorescence images of a mouse administered 10 nmol of a peptide of SEQ ID NO: 108 conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A) at 24 hours post-administration. FIG. 10A illustrates an image of a frozen section of a mouse, 24 hours after administration of 10 nmol of a peptide of SEQ ID

NO: 108 conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A). FIG. 10B illustrates the fluorescence signal in the mouse, corresponding to the section shown in FIG. 10A, 24 hours after administration of 10 nmol of a peptide of SEQ ID NO: 108 conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A). FIG. IOC illustrates an image of a different frozen section of the mouse, 24 hours after administration of 10 nmol of a peptide of SEQ ID NO: 108 conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A). FIG. 10D illustrates the fluorescence signal in the mouse, corresponding to the section shown in FIG. IOC, 24 hours after administration of 10 nmol of a peptide of SEQ ID NO: 108 conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A). FIG. 10E illustrates an image of a different frozen section of the mouse, 24 hours after administration of 10 nmol of a peptide of SEQ ID NO: 108 conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A). FIG. 10F illustrates a fluorescence signal in the mouse, corresponding to the section shown in FIG. 10E, 24 hours after administration of 10 nmol of a peptide of SEQ ID NO: 108 conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A).

[0443] FIG. 13 shows IVIS fluorescence imaging of an isolated hind limb from a first mouse and an isolated hind limb from a second mouse after administration of 10 nmol SEQ ID NO: 108 peptide conjugated to a Cy5.5 fluorophore (SEQ ID NO: 108A). FIG. 13A shows the right hind limb with skin removed from a first mouse and from a second mouse 3 hours after peptide administration. FIG. 13B shows the right hind limb with muscle removed from a first mouse and from a second mouse 3 hours after peptide administration. FIG. 13C shows the right hind limb with skin removed from a first mouse and from a second mouse 24 hours after peptide administration. FIG. 13D shows the right hind limb with muscle removed from a first mouse and from a second mouse 24 hours after peptide administration. FIG. 13E shows the right hind limb with skin removed from a first mouse and from a second mouse 48 hours after peptide administration. FIG. 13F shows the right hind limb with muscle removed from a first mouse and from a second mouse 48 hours after peptide administration. FIG. 13G shows the right hind limb with skin removed from a first mouse and from a second mouse 72 hours after peptide administration. FIG. 13H shows the right hind limb with muscle removed from a first mouse and from a second mouse 72 hours after peptide administration. Peptide fluorescence was observed in the knee joints of isolated right hind limbs at all time points tested.

EXAMPLE 86

Peptide Resistance Under Various Conditions

[0444] This example illustrates peptide stability under various stress conditions such as high temperature, low pH, reducing agents, and proteases. To determine resistance to high

temperatures, cystine-dense peptides (CDPs) were incubated at 0.5 mM in PBS at 75°C or 100°C for 1 h and pelleted, and the supernatant was analyzed with reversed-phase chromatography (RPC). To determine resistance to proteolytic digestion, CDPs were mixed with 50 U of porcine pepsin, in simulated gastric fluid at pH 1.0, or 50 U of porcine trypsin in PBS, incubated for 30 minutes at 37 °C and analyzed with RPC. Oxidized and reduced forms (prepared through addition 10 mM DTT) were compared. Circular Dichroism spectroscopy was used in order to measure the secondary structure of peptides with a Jasco J-720W spectropolarimeter in a cell with a 1.0-mm path length, and CDPs were diluted into 20 mM phosphate buffer, pH 7.4, at a concentration of 15-25 μΜ. These conditions were expected to denature or degrade conventional globular proteins and many peptides. In TABLE 11, "high" resistance indicated a high amount of the peptide remained or was retained as unmodified under the given experimental conditions and "low" resistance indicated a low amount of the peptide remained or was retained unmodified under the given experimental conditions. Notably, the experimental conditions described in this example were more extreme stress conditions than to many standard in vivo or physiologic conditions, in vitro conditions, conditions during manufacturing, and handling conditions. As such, even "low" resistance can indicate meaningful resistance to these stress conditions that may have applicability for a number of uses described herein. The data from these studies are shown in TABLE 11. The peptides tested, SEQ ID NO: 315, SEQ ID NO: 317 and SEQ ID NO: 482, showed high resistance to one or more of the conditions tested.

TABLE 11 - Resistance of SEQ ID NO: 317, SEQ ID NO: 315, and SEQ ID NO: 482 to

Various Conditions

[0445] While certain embodiments of the present disclosure have been exemplified or shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. It is not intended that the disclosure be limited by the specific examples provided within the specification. While the disclosure has been described with reference to the aforementioned specification, the descriptions and illustrations of the

embodiments herein are not meant to be construed in a limiting sense. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the disclosure. Furthermore, it shall be understood that all embodiments of the disclosure are not limited to the specific depictions, configurations or relative proportions set forth herein which depend upon a variety of conditions and variables. It should be understood that various alternatives to the embodiments of the disclosure described herein may be employed in practicing the disclosure. It is therefore contemplated that the disclosure shall also cover any such alternatives, modifications, variations or equivalents. It is intended that the following claims define the scope of the disclosure and that methods and structures within the scope of these claims and their equivalents be covered thereby.