BACKGROUND

Field

[0001] The present disclosure relates generally to the field of cancer treatment and more specifically to the field of treatments for ocular melanoma.

Background

[0002] Ocular melanoma (OM) is a primary cancer of melanocytes within the eye. Distinct from cutaneous melanoma, OM represents approximately 5% of all melanomas and affects roughly 2,500 adults every year in the U.S, with an incidence of roughly 21 cases per million people per year over age 50. While relatively rare, OM has been associated with light eye color, increased age, certain genetic skin disorders (such as dysplastic naevus syndrome), and cumulative UV exposure. OM is an aggressive form of cancer which by itself has symptoms and complications including glaucoma or other disorders of intraocular pressure, eye pain, blurred vision, vision changes and vision loss. OM may affect any of the three main areas of the eye, including the iris, the ciliary body, or the choroid or posterior uvea, or any combination of the three. OM may also develop in the conjunctiva. OM frequently metastasizes to the liver, as well as to the lung, bones, and other sites. Survival of patients with metastatic OM is low, with liver metastases being almost universally fatal, even when the primary eye tumor is treated successfully. Thus, there is a need for treatments for both tumors in situ, and for metastases of, ocular melanoma.

SUMMARY

[0003] In some embodiments according to the methods and compositions described herein, the present disclosure provides a method of treating ocular melanoma, including symptoms, sequelae, and metastases thereof, including micrometastases, by selecting a subject in need of treatment for ocular melanoma, and then by administering to said subject a therapeutically effective amount of a composition comprising ranpirnase. In some embodiments, said subject has, shows signs and/or symptoms of, or suffers from ocular melanoma. In some embodiments, selecting a subject in need of treatment for ocular melanoma, including symptoms, sequelae, and metastases thereof, including micrometastases, may further comprise determining the presence of mutations in the BRAF, NRAS and/or KIT genes or any combination thereof. In some embodiments, said ocular melanoma is a tumor in situ, a circulating tumor, a metastatic tumor, a micrometastatic tumor, or a mass or neoplasia derived from an ocular melanoma.

[0004] In some embodiments, the composition to be administered may further comprise one or more additional active compounds, such as, but not limited to, an immunotherapy, a checkpoint inhibitor, a kinase inhibitor, a cytoskeletal inhibitor, or a chemotherapeutic agent, or any combination thereof. In some embodiments, the composition to be administered may further comprise one or more additional nucleases. In some embodiments, the methods as disclosed herein may further comprise separately administering one or more additional nucleases, or an immunotherapy, a checkpoint inhibitor, a kinase inhibitor, a cytoskeletal inhibitor, or a chemotherapeutic agent, or any combination thereof. In some embodiments, said further administration may be prior to, coincident with, or following the administration of any composition comprising ranpirnase. In some embodiments, the composition is administered intraocularly, opthalmically, or parenterally, and may be administered by suprachoroidal injection or suprachoroidal implantation. In some embodiments, the composition is administered by subchoroidal injection or subchoroidal implantation. In some embodiments, the composition is administered by injection or implantation to the orbital or periorbital space. In some embodiments, the composition is administered by intravenous or intraperitoneal infusion. In some embodiments, further administration of any additional nucleases, or an immunotherapy, a checkpoint inhibitor, a kinase inhibitor, a cytoskeletal inhibitor, or a chemotherapeutic agent, or any combination thereof is by by intravenous or intraperitoneal infusion, venous intrahepatic infusion, arterial intrahepatic infusion, or infusion via the biliary ducts. In some embodiments, administration of any additional or subsequent dosages of any composition as described herein is by intravenous or intraperitoneal infusion, venous intrahepatic infusion, arterial intrahepatic infusion, or infusion via the biliary ducts. [0005] In some embodiments, the methods and compositions of the present disclosure may further comprise a brachytherapy or radiation beam therapy.

[0006] In some embodiments, the composition to be administered may further comprise an antibody, an epitope binding fragment of an antibody, a T-cell receptor, a chimeric T-cell receptor or a CAR-T cell.

[0007] In some embodiments, the ranpimase may be encapsulated within, conjugated to, fused to, associated with, or encoded by, an oncolytic virus.

[0008] In some embodiments, the ranpimase may administered genetically, such as by administration of a nucleic acid such as a cDNA, genomic DNA, viral DNA, mRNA, viral RNA, or other such nucleic acid as may be administrable to a subject and which can encode a polypeptide with at least 85%, 90%, 95%, or 100% sequence identity to the polypeptide disclosed in SEQ ID NOs: 1-3. In some embodiments, said nucleic acid may comprise a nucleic acid with at least 85%, 90%, 95%, or 100% sequence identity to the sequence disclosed in SEQ ID NO: 4.

DETAILED DESCRIPTION

[0009] Ranpimase is a Type A ribonuclease derived from Rana pipiens oocytes.

Particular activities of ranpimase are disclosed in U.S. Patents No. 9,919,034, 9,642,794, ,

8,808,690, , 8,663,964, 8,518,399, 7,763,449, 7,585,655, 7,585,654, 7,556,953, 7,556,952, 7,556,951, 7,473,542, 7,442,536, 7,442,535, 7,229,824, 6,423,515, 6,290,951, 6,239,257,

6,175,003, 5,595,734, 5,540,925, and 5,529,775, which are hereby incorporated by reference with respect to their disclosure of compositions and methods incorporating, utilizing, or rendering comparisons to ranpimase.

[0010] In some embodiments, the methods and compositions of the present disclosure provide for the administration of ranpimase to a subject. In some embodiments, said subject is one who has contracted, suffers from, or exhibits one or more signs and/or symptoms of, a disease or disorder. In some embodiments, said disease or disorder is a disease or disorder of the eye.

[0011] In some embodiments, the disease or disorder to be treated comprises a cancer or neoplasia. In some embodiments, said cancer or neoplasia is a melanoma. In some embodiments, said melanoma is an ocular melanoma. In some embodiments, said ocular melanoma is a corneal melanoma, a uveal melanoma, or a conjunctival melanoma. In some embodiments, a uveal melanoma may comprise one or more cancers of the choroid, iris, and/or ciliary body. As used herein, a cancer or neoplasia, including a melanoma or ocular melanoma, may further comprise tumors in situ, circulating cells thereof, and/or metastases thereof. In some embodiments, said metastases include metastases to the liver.

[0012] In some embodiments, the methods as disclosed herein further comprise genetic characterization of one or more ocular melanomas, ocular tumors, metastases, tumors in situ, and/or genetic characterization and/or genotyping of the somatic cells of a subject. In some embodiments, said subject has, shows symptoms of, or is at risk of, an ocular condition which may comprise ocular melanoma. In some embodiments, said one or more ocular melanomas, ocular tumors, metastases, tumors in situ, and/or somatic cells may contain one or more mutations may comprise one or more of a substitution, deletion, insertion, translocation, inversion, and/or chromosomal break or chromosomal rearrangement, or other such alterations as are known to or may affect the expression, localization, or function, including physiological function, of a protein, RNA, or other gene product. In some embodiments, said one or more mutations may affect coding regions, noncoding regions, splice sites, regulatory regions, or other portions of a gene or genetic site as is known to or may affect the expression, localization, or function, including physiological function, of a protein, RNA, or other gene product. In some embodiments, said one or more ocular melanomas, ocular tumors, metastases, tumors in situ, and/or somatic cells may contain one or more mutations of the Bapl gene. In some embodiments, said one or more ocular melanomas, ocular tumors, metastases, tumors in situ, and/or somatic cells may contain mutations as disclosed herein of one or more of the BRAF, NRAS and/or KGG genes or any combination thereof. In some embodiments, said one or more ocular melanomas, ocular tumors, metastases, tumors in situ, and/or somatic cells may lack mutations as disclosed herein of one or more of the BRAF, NRAS and/or KGG genes or any combination thereof.

[0013] In some embodiments, the methods of the present disclosure provide for the administration of one or more enzymes, proteins, peptides, peptidomimetics, peptide fusions, peptide conjugates, compositions comprising one or more compounds associated with a peptide (“associates”), or other agents as described herein which may be efficacious in the treatment of diseases or disorders of or originating in the eye, such as cancers or neoplasias, which may comprise an ocular melanoma as described herein, or one or more symptoms, complications, sequelae, and.or metastases thereof. In some embodiments, said one or more enzymes, proteins, peptides, peptidomimetics, peptide fusions, peptide conjugates, associates, or other agents as described herein may comprise a nuclease. In some embodiments, said nuclease may comprise a ribonuclease or RNase. In some embodiments, said RNase may comprise an RNase A, RNase H, RNase T, RNase IP, RNase L, RNase P, RNase Tl, RNase T2, RNase U2, RNase V, RNase PH, RNase D, RNase T, RNase PhyM, RNase R, an oligoribonuclease, and exoribonuclease I or II, and/or a polynucleotide phosphorylase, and/or any combination thereof, or any other enzyme, protein, peptide, catalyst, or other agent which may induce the degradation, destruction, or covalent modification of one or more ribonucleic acid molecules (RNAs). In some embodiments, said RNAs may comprise one or more of a messenger RNA, transfer RNA, ribosomal RNA, noncoding RNA, small inhibitory RNA, long noncoding RNA, regulatory RNA, viral RNA, noncoding RNA, and/or any other such RNA as may be present in, localized to, or co- occurrent with an ocular melanoma and/or other cancers, neoplasias, metastases, or conditions as described herein. In some embodiments, said RNase comprises an RNase A. In some embodiments said RNase comprises, consists essentially of, or consists of, ranpirnase. In some embodiments, said RNase comprises, consists of, or consists essentially of, a nucleic acid, such as a genomic DNA, plasmid, cDNA, mRNA, viral RNA, or the like, which encodes an RNase A or one or more enzymes or ribozymes having RNase A activity. In some embodiments, said RNase comprises, consists of, or consists essentially of, a nucleic acid, such as a genomic DNA, plasmid, cDNA, mRNA, viral RNA, or the like, which encodes ranpirnase.

[0014] In some embodiments, the compositions to be administered according to the methods of the present disclosure may further comprise one or more excipients, such as a carrier, diluent, adhesive, bulking agent, solvent, tonicity adjuster, buffer, preservative, or other excipient as is known in the art of pharmaceutical formulation. In some embodiments, said compositions to be administered may be formulated for ocular or ophthalmic administration. In some embodiments, said ocular or ophthalmic formulations may comprise one or more of an eyedrop, salve, cream, ointment, a composition for intravitreal injection, composition for suprachoroidal injection, and/or a depot form, including but not limited to forms for injection into the choroidal, uveal, or orbital spaces, a such as liquid forms, gel forms, hydrogels, and or patches, such as resorbable patches. Said depot forms may comprise any such forms as are known in the art. Exemplary forms may comprise poly lactic acid, polylactic-glycolic acid, collagen, hyaluronic acid, hyaluronan, chitin, chitosan, starch, polysaccharide, and/or other compositions as are known in the art for the formation of gels, hydrogels, patches, fabrics, or other forms as may be suitable for administration of agents to the eye. In some embodiments, said forms may include fibers, electro spun fibers, nanofibers, microfibers, particles, microparticles, nanoparticles, spray-dried forms, or other forms as are known in the preparation of compositions for administration to the eye. In some embodiments according to the compositions and methods as described herein, said compositions may be administered through the use of a port, implantable or external pump, or implantable or external reservoir. In some embodiments, said compositions are formulated for systemic administration.

[0015] In some embodiments, the compositions to be administered according to the methods of the present disclosure comprise ranpirnase and/or amphinase. In some further embodiments, said compositions further comprise one or more additional chemotherapeutic or anti-cancer agents, such as an immunotherapy, a checkpoint inhibitor, a kinase inhibitor, a cyto skeletal inhibitor, or a chemotherapeutic agent, or any combination thereof. Exemplary chemotherapeutic or anti-cancer agents, including immunotherapies, checkpoint inhibitors, cytoskeletal inhibitors, or kinase inhibitors which may be combined with or coadministered with ranpirnase according to the present methods include but are not limited to paclitaxel, vincristine, vinblastine, mertansine, epothilone, docetaxel, discodermolide, combrestatin, podophyllotoxin, CI-980, phenylahistins, steganacins, curacins, 2-methoxy estradiol, E7010, methoxy benzenesuflonamides, vinorelbine, vinflunine, vindesine, dolastatins, spongistatin, rhizoxin, tasidotin, halichondrins, hemiasterlins, cryptophycin 52, MMAE, eribulin mesylate, icin, abrin, ribonuclease (RNase), DNase I, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin, diphtheria toxin, Pseudomonas exotoxin, Pseudomonas endotoxin, a cytokine, a stem cell growth factor, a lymphotoxin, a hematopoietic factor, a colony stimulating factor (CSF), an interferon (IFN), erythropoietin, thrombopoietin and a combination thereof. Specifically useful are lymphotoxins such as tumor necrosis factor (TNF), hematopoietic factors, such as interleukin (IL), colony stimulating factor, such as granulocyte-colony stimulating factor (G-CSF) or granulocyte macrophage-colony stimulating factor (GM-CSF), interferon, such as interferons-. alpha., -.beta., -.gamma or - .lamda., and stem cell growth factor, such as that designated“Sl factor”. Included among the cytokines are growth hormones such as human growth hormone, N-methionyl human growth hormone, and bovine growth hormone; parathyroid hormone; thyroxine; insulin; proinsulin; relaxin; prorelaxin; glycoprotein hormones such as follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH), and luteinizing hormone (LH); hepatic growth factor; prostaglandin, fibroblast growth factor; prolactin; placental lactogen, OB protein; tumor necrosis factor-. alpha and -.beta.; mullerian-inhibiting substance; mouse gonadotropin- associated peptide; inhibin; activin; vascular endothelial growth factor; integrin; thrombopoietin (TPO); nerve growth factors such as NGF-.beta.; platelet-growth factor; transforming growth factors (TGFs) such as TGF-. alpha and TGF-.beta.; insulin-like growth factor-I and -II; erythropoietin (EPO); osteoinductive factors; interferons such as interferon- . alpha., -.beta., -.gamma and -.lamda.; colony stimulating factors (CSFs) such as macrophage-CSF (M-CSF); interleukins (ILs) such as IL-l, IL-l. alpha., IL-2, IL-3, IL-4, IL- 5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-l l, IL-12; IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-21, IL-25, LIF, kit-ligand or FLT-3, angiostatin, thrombospondin, endostatin, tumor necrosis factor and lymphotoxin (LT), idelalisib, Wortmannin, demethoxyviridin, perifosine, PX-866, IPI-145 (duvelisib), BAY 80-6946, BEZ235, RP6530, TGR1202, SF1126, INK1117, GDC- 0941, B KM 120, XL147, XL765, Palomid 529, GSK1059615, ZSTK474, PWT33597, IC87114, TG100-115, CAL263, PI-103, GNE477, CUDC-907, AEZS-136, LY294002, ibrutinib (PCI-32765), PCI-45292, CC-292 (AVL-292), ONO-4059, GDC-0834, LFM-A13 RN486, olaparib, talazoparib (BMN-673), rucaparib, veliparib, CEP 9722, MK 4827, BGB- 290, ABT-888, AG014699, BSI-201, CEP-8983, 3-aminobenzamide, 5-fluorouracil, afatinib, aplidin, azaribine, anastrozole, anthracyclines, axitinib, AVL-101, AVL-291, bendamustine, bleomycin, bortezomib, bosutinib, bryostatin-l, busulfan, calicheamycin, camptothecin, carboplatin, lO-hydroxycamptothecin, carmustine, celebrex, chlorambucil, cisplatin (CDDP), Cox-2 inhibitors, irinotecan (CPT-l l), SN-38, carboplatin, cladribine, camptothecans, crizotinib, cyclophosphamide, cytarabine, dacarbazine, dasatinib, dinaciclib, docetaxel, dactinomycin, daunorubicin, doxorubicin, 2-pyrrolinodoxorubicine (2P-DOX), cyano- morpholino doxorubicin, doxorubicin glucuronide, epirubicin glucuronide, erlotinib, estramustine, epidophyllotoxin, erlotinib, entinostat, estrogen receptor binding agents, etoposide (VP 16), etoposide glucuronide, etoposide phosphate, exemestane, fingolimod, floxuridine (FUdR), 3',5'-P-dioleoyl-FudR (FUdR-dO), fludarabine, flutamide, famesyl- protein transferase inhibitors, flavopiridol, fostamatinib, ganetespib, GDC-0834, GS-1101, gefitinib, gemcitabine, hydroxyurea, ibrutinib, idarubicin, idelalisib, ifosfamide, imatinib, L- asparaginase, lapatinib, lenolidamide, leucovorin, LFM-A13, lomustine, mechlorethamine, melphalan, mercaptopurine, 6-mercaptopurine, methotrexate, mitoxantrone, mithramycin, mitomycin, mitotane, navelbine, neratinib, nilotinib, nitrosurea, olaparib, plicomycin, procarbazine, paclitaxel, PCI-32765, pentostatin, PSI-341, raloxifene, semustine, sorafenib, streptozocin, SU11248, sunitinib, tamoxifen, temazolomide (an aqueous form of DTIC), transplatinum, thalidomide, thioguanine, thiotepa, teniposide, topotecan, uracil mustard, vatalanib, vinorelbine, vinblastine, vincristine, vinca alkaloids, nivolumab, pembrolizumab, trametanib, or ZD 1839, or any combination thereof.

[0016] In some embodiments, the methods as described herein further comprise a brachytherapy or radiation beam therapy. Exemplary radiation beam therapies may include administration of gamma radiation, alpha particles, protons, or other radiation as may be known by one of skill in the art. Exemplary brachytherapies include but are not limited to implantation of seeds containing ¹²⁵ I, ¹³¹ Cs, ¹³⁷ Cs, ⁶⁰ Co, ¹⁹² Ir, ¹⁰³ Pd, ¹⁰⁶ Ru, and/or ²²⁶ Ra, or other such radiation sources as are or may be known to one of skill in the art. Implantation of seeds for brachytherapy may be intravitreal, macular, retinal, subretinal, conjunctival, comeal, within the orbital space, or elsewhere as dictated by the source, the location of the tumor, and/or any other considerations as may be considered relevant by the individual rendering treatment and/or by one of skill in the art.

[0017] As noted elsewhere herein, the present administration contemplate coadministration of ranpimase and/or amphinase with one or more additional active compounds within or during the course of treatment of ocular melanoma. Said coadministration may be sequential, such as prior to, or following, the administration of ranpirnase, or may be simultaneous. As used herein,“coadministration” further encompasses formulation of a compound with ranpirnase in a single dosage form, as well as administration of a compound as a separate dosage form. Exemplary compounds for coadministration with ranpirnase may further comprise one or more small molecules, peptides, peptoids, peptide nucleic acids (PNAs), nucleic acids including DNA, RNA, and modified RNA such as, but not limited to, antisense RNA, siRNA, or lncRNA. Said compounds may also comprise an antibody or fragment thereof, especially an epitope binding fragment thereof or other binding protein capable of binding to any target known to be expressed by, secreted by, or utilized within or on the surface of, ocular melanoma cells or migrating, circulating, or metastatic cells of an ocular melanoma. Exemplary antibodies or binding fragments thereof may comprise polyclonal antibodies, monoclonal antibodies, single chain variable fragments (ScFvs), single chain antibodies, or the like. Other representative compositions as contemplated herein may further comprise T-cell receptors, synthetic T-cell receptors, binding peptides, and the like, whether alone or incorporated into a cell, such as a CAR-T cell. Said antibodies, antibody fragments, and other binding proteins may be produced by any method as is known in the art, including in vivo methods, hybridoma methods, recombinant methods, phage display, total synthesis, or other methods as are known in the art. The present disclosure also contemplates the administration of one or more aptamers, which may comprise nucleic acids and nucleic acid derivatives such as DNA, RNA, peptide nucleic acids (PNAs), and modified RNA such as, but not limited to, antisense RNA, siRNA, or lncRNA and the like, or derivatives thereof.

[0018] In some embodiments according to the present disclosure, ranpirnase may be conjugates with, fused to, or noncovalently associated with one or more additional active compounds. Exemplary compounds for conjugation, fusion, or association with ranpirnase may further comprise one or more immunotherapies, checkpoint inhibitors, a kinase inhibitors, a cytoskeletal inhibitors, or a chemotherapeutic agents as disclosed elsewhere herein, one or more small molecules, peptides, peptoids, peptide nucleic acids (PNAs), nucleic acids including DNA, RNA, and modified RNA such as, but not limited to, antisense RNA, siRNA, or lncRNA, or any combination thereof. Said compounds may also comprise an antibody or fragment thereof, especially an epitope binding fragment thereof or other binding protein capable of binding to any target known to be expressed by, secreted by, or utilized within or on the surface of, ocular melanoma cells or migrating, circulating, or metastatic cells of an ocular melanoma. Exemplary antibodies or binding fragments thereof may comprise polyclonal antibodies, monoclonal antibodies, single chain variable fragments (ScFvs), single chain antibodies, or the like. As described elsewhere herein, other representative compositions as contemplated herein may further comprise T-cell receptors, synthetic T-cell receptors, binding peptides, and the like, whether alone or incorporated into a cell, such as a CAR-T cell. Said antibodies, antibody fragments, and other binding proteins may be produced by any method as is known in the art, including in vivo methods, hybridoma methods, recombinant methods, phage display, total synthesis, or other methods as are known in the art. The present disclosure also contemplates the conjugation, fusion, or association of ranpimase with one or more aptamers, which may comprise nucleic acids and nucleic acid derivatives such as DNA, RNA, peptide nucleic acids (PNAs), and modified RNA such as, but not limited to, antisense RNA, siRNA, or lncRNA and the like, or derivatives thereof.

[0019] In some embodiments, ranpirnase may be delivered genetically, such as by administration of a nucleic acid, such as a DNA or RNA molecule encoding ranpirnase, which may be provided as part of a larger genetic element such as a plasmid, artificial chromosome, or viral genome including a synthetic viral genome. Said nucleic acid may comprise a nucleic acid that encodes a polypeptide that is 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% identical to SEQ ID NOs: 1-3. Said administration may further comprise the administration of one or more genetically modified cells such as stem cells or modified immune cells, which may be of autologous or heterologous origin with respect to the subject to which they are administered. Said cells may, in some embodiments, express ranpirnase internally. Said cells may, in some embodiments, secrete ranpirnase.

[0020] In some embodiments, ranpirnase may be encapsulated within, conjugated to, fused to, associated with, or encoded by, an oncolytic virus. Representative oncolytic viruses may include one or more of a vaccinia virus, a vesicular stomatitis virus, a poliovirus, a reovirus, a senecavirus, a Semliki Forest virus, an adenovirus, or the like, including naturally occurring, engineered, and artificial variants thereof, or any hybrid, non-naturally occurring, artificial, or engineered virus delivery vehicle as are or may be known in the art. Said viruses may encode such proteins as are necessary for viral function or for attenuated or limited viral function consistent with oncolytic activity, in addition to containing ranpimase either as a protein conjugated to, fused to, or associated with the virion or envelope, or encoding one or more nucleic acids as described herein, comprising a nucleic acid that encodes a polypeptide that is 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% identical to SEQ ID NOs: 1-3. Said nucleic acid may also comprise a nucleic acid having 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% identityto SEQ ID NO: 4.

[0021] Administration of the agents disclosed herein can be via any of the accepted modes of administration for agents that serve similar utilities including, but not limited to, orally, subcutaneously, intravenously, intranasally, topically, transdermally, intraperitoneally, intrathecally, intrahepaticcally, intramuscularly, intrapulmonarilly, vaginally, rectally, ophthalmically, or intraocularly.

[0022] The compositions described herein may be provided in unit dosage form. As used herein, a“unit dosage form” is a composition containing an amount of agent that is suitable for administration to a subject, in a single dose, according to good medical practice. The preparation of a single or unit dosage form however, does not imply that the dosage form is administered once per day or once per course of therapy. A unit dosage form may comprise a single daily dose or a fractional sub-dose wherein several unit dosage forms are to be administered over the course of a day in order to complete a daily dose. According to the present disclosure, a unit dosage form may be given more or less often that once daily, and may be administered more than once during a course of therapy. Such dosage forms may be administered in any manner consistent with their formulation, including orally, ophthalmically, intraocularly, parenterally, and may be administered as an infusion over a period of time (e.g., from about 30 minutes to about 2-6 hours). While single administrations are specifically contemplated, the compositions administered according to the methods described herein may also be administered as a continuous infusion or via an implantable infusion pump.

[0023] A liquid composition for topical ophthalmic use is formulated such that it can be administered topically to the eye. Said composition may comprise an immediate release form, a controlled release form, or a depot form. Patient comfort may be maximized as much as possible, although sometimes formulation considerations (e.g. drug stability) may necessitate less than optimal comfort. In the case that comfort cannot be maximized, the liquid may be formulated such that the liquid is tolerable to the patient for topical ophthalmic use. Additionally, an ophthalmically acceptable liquid may either be packaged for single use, or contain a preservative to prevent contamination over multiple uses.

[0024] A liquid composition or depot form may further be formulated for intraocular injection, such as intravitreal injection using such carriers and excipients as are known in the art. Further, liquid compositions may be formulated for injection into the orbital space, into optic arteries, veins, or capillaries, into the optic nerve channel, or within the optic nerve sheath.

[0025] Further liquid compositions or depo forms may be formulated for suprachoroidal injection. Such compositions may be injected, such as with a microinjector, through the sclera into the suprachoroidal space.

[0026] For ophthalmic application, solutions or medicaments are often prepared using a physiological saline solution as a major vehicle. Ophthalmic solutions may preferably be maintained at a comfortable pH with an appropriate buffer system. The formulations may also contain conventional, pharmaceutically acceptable preservatives, stabilizers, compendial excipients, viscosity additives and/or viscosity modifiers, and surfactants.

[0027] Preservatives that may be used in the pharmaceutical compositions disclosed herein include, but are not limited to, benzalkonium chloride, PHMB, chlorobutanol, thimerosal, phenylmercuric, acetate and phenylmercuric nitrate. A useful surfactant is, for example, Tween 80. Likewise, various useful vehicles may be used in the ophthalmic preparations disclosed herein. These vehicles include, but are not limited to, polyvinyl alcohol, povidone, hydroxypropyl methyl cellulose, poloxamers, carboxymethyl cellulose, hydroxyethyl cellulose and purified water.

[0028] Tonicity adjustors may be added as needed or convenient. They include, but are not limited to, salts, particularly sodium chloride, potassium chloride, mannitol and glycerin, or any other suitable ophthalmically acceptable tonicity adjustor.

[0029] Various buffers and means for adjusting pH may be used so long as the resulting preparation is ophthalmically acceptable. For many compositions, the pH will be between 4 and 9. Accordingly, buffers include acetate buffers, citrate buffers, phosphate buffers and borate buffers. Acids or bases may be used to adjust the pH of these formulations as needed.

[0030] Ophthalmically acceptable antioxidants include, but are not limited to, sodium metabisulfite, sodium thiosulfate, acetylcysteine, butylated hydroxyanisole and butylated hydroxy toluene.

[0031] Other excipient components, which may be included in the ophthalmic preparations, are chelating agents. A useful chelating agent is edetate disodium, although other chelating agents may also be used in place or in conjunction with it.

[0032] For intravenous administration, the compositions described herein may be dissolved or dispersed in a pharmaceutically acceptable diluent, such as a saline or dextrose solution. Suitable excipients may be included to achieve the desired pH, including but not limited to NaOH, sodium carbonate, sodium acetate, HC1, and citric acid. In various embodiments, the pH of the final composition ranges from 2 to 9, or preferably from 4 to 7.5. Antioxidant excipients may include sodium bisulfite, acetone sodium bisulfite, sodium formaldehyde, sulfoxylate, thiourea, and EDTA. Other non-limiting examples of suitable excipients found in the final intravenous composition may include sodium or potassium phosphates, citric acid, tartaric acid, gelatin, and carbohydrates such as dextrose, mannitol, and dextran. Further acceptable excipients are described in Powell, et al., Compendium of Excipients for Parenteral Formulations, PDA J Pharm Sci and Tech 1998, 52 238-311 and Nema et al., Excipients and Their Role in Approved Injectable Products: Current Usage and Future Directions, PDA J Pharm Sci and Tech 2011, 65 287-332, both of which are incorporated herein by reference in their entirety. Antimicrobial agents may also be included to achieve a bacteriostatic or fungistatic solution, including but not limited to phenylmercuric nitrate, thimerosal, benzethonium chloride, benzalkonium chloride, phenol, cresol, and chlorobutanol.

[0033] Compositions for intravenous administration may be provided to caregivers in the form of one more solids that are reconstituted with a suitable diluent such as sterile water, saline or dextrose in water shortly prior to administration. In other embodiments, the compositions are provided in solution ready to administer parenterally. In still other embodiments, the compositions are provided in a solution that is further diluted prior to administration. In embodiments that include administering a combination of a compound described herein and another agent, the combination may be provided to caregivers as a mixture, or the caregivers may mix the two agents prior to administration, or the two agents may be administered separately.

[0034] The actual unit dose of the active compounds described herein depends on the specific composition, the preferred route of administration, and on the status of the cancer or neoplasia to be treated. For example, in some embodiments, especially those incorporating intravenous infusion and/or those intended to treat distant metastases, the dose may be from about O.OOlmg/kg to about 100 mg/kg or more of body weight, from 0.01 mg/kg to about 120 mg/kg or more of body weight, from about 0.05 mg/kg or less to about 70 mg/kg, from about 0.1 mg/kg to about 50 mg/kg of body weight, from about 1.0 mg/kg to about 10 mg/kg of body weight, from about 5.0 mg/kg to about 10 mg/kg of body weight, or from about 10.0 mg/kg to about 20.0 mg/kg of body weight. In some embodiments, the dose may be less than 100 mg/kg, 90 mg/kg, 80 mg/kg, 70 mg/kg, 60 mg/kg, 50 mg/kg, 40 mg/kg, 30 mg/kg, 25 mg/kg, 20 mg/kg, 10 mg/kg, 8 mg/kg, 7.5 mg/kg, 6 mg/kg, 5 mg/kg, 4 mg/kg, 3 mg/kg, 2.5 mg/kg, 1 mg/kg, 0.5mg/kg, 0.1 mg/kg, 0.05 mg/kg or 0.005 mg/kg of body weight. In some embodiments, the actual unit dose is 0.005, 0.05, 0.07, 0.1, 0.3, 1.0, 3.0, 5.0, 7.0, 10.0 or 25.0 mg/kg of body weight. Thus, for administration to a 70 kg person, the dosage range would be from about 0.1 mg to 70 mg, from about 1 mg to about 50 mg, from about 0.5 mg to about 10 mg, from about 1 mg to about 10 mg, from about 2.5 mg to about 30 mg, from about 35 mg or less to about 700 mg or more, from about 7 mg to about 600 mg, from about 10 mg to about 500 mg, from about 20 mg to about 300 mg, from about 600 mg to about 1200 mg, or from about 200 mg to about 2000 mg. In some embodiments, the actual unit dose is 5 mg. In some embodiments the actual unit dose is 10 mg. In some embodiments, the actual unit dose is 25 mg. In some embodiments, the actual unit dose is 1500 mg or less. In some embodiments, the actual unit dose is 1250 mg or less. In some embodiments, the actual unit dose is 1000 mg or less. In some embodiments, the actual unit dose is 750 mg or less. In some embodiments, the actual unit dose is 500 mg or less. In some embodiments, the actual unit dose is 250 mg or less.

[0035] In some embodiments, a unit dose may be administered daily, or less often than daily, or more often than daily, as may be determined by one of skill in the art of treating ophthalmic cancers or their symptoms, sequelae, and/or metastases. In some embodiments, a unit dose may be administered daily. In some embodiments, a unit dose may be administered less often than daily. In some embodiments, a unit dose may be administered more often than daily. In some embodiments, a unit dose may be administered 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times per day. In some embodiments, a unit dose may be administered more than 10 times per day. In some embodiments a unit does may be administered ever other day. In some embodiments, a unit dose may be administered every third day. In some embodiments, a unit dose may be administered every 3, 4, 5, 6, 7, 8, 9, or 10 days. In some embodiments, a unit dose may be administered less often than every 10 days. In some embodiments, the dosing interval may be varied according to a schedule to be determined by one of skill in the art of treating ophthalmic cancers or their symptoms, sequelae, and/or metastases. In some embodiments, the dosing interval may be varied according to observations related to the health of the patient, the response of the patient to the administration of the compositions described herein, or the presence, absence, and/or progression of one or more ophthalmic cancers or their symptoms, sequelae, and/or metastases.

Definitions

[0036] The term “mammal” is used in its usual biological sense. Thus, it specifically includes humans and non-human mammals such as dogs, cats, horses, donkeys, mules, cows, domestic buffaloes, camels, llamas, alpacas, bison, yaks, goats, sheep, pigs, elk, deer, domestic antelopes, and non-human primates as well as many other species. [0037] “Subject” as used herein, means a human or a non-human mammal including but not limited to a dog, cat, horse, donkey, mule, cow, domestic buffalo, camel, llama, alpaca, bison, yak, goat, sheep, pig, elk, deer, domestic antelope, or a non-human primate selected for treatment or therapy.

[0038] “Subject suspected of having” means a subject exhibiting one or more clinical indicators of a disease or condition. In certain embodiments, the disease or condition is a cancer or neoplasia. In certain embodiments, the disease or condition is ocular melanoma, or any sequela or metastasis thereof.

[0039] “Subject in need thereof’ means a subject identified as in need of a therapy or treatment.

[0040] A therapeutic effect relieves, to some extent, one or more of the symptoms of a disease or disorder, and includes curing the disease or disorder.“Curing” means that the symptoms of active disease are eliminated. However, certain long-term or permanent effects of the disease may exist even after a cure is obtained (such as extensive tissue damage).

[0041] “Treat,”“treatment,” or“treating,” as used herein refers to administering a pharmaceutical composition for prophylactic and/or therapeutic purposes. The term “prophylactic treatment” refers to treating a patient who does not yet have the relevant disease or disorder, but who is susceptible to, or otherwise at risk of, a particular disease or disorder, whereby the treatment reduces the likelihood that the patient will develop the disease or disorder. The term“therapeutic treatment” refers to administering treatment to a patient already having a disease or disorder.

[0042] “Preventing” or“prevention” refers to delaying or forestalling the onset, development or progression of a condition or disease for a period of time, including weeks, months, or years.

[0043] As used herein,“migrating cancer” refers to a cancer in which the cancer cells forming the tumor, regardless of a large or small (micrometastatic) cluster, migrate and subsequently grow as malignant implants at a site other than the site of the original tumor. The cancer cells migrate via seeding the surface of the peritoneal, pleural, pericardial, or subarachnoid spaces to spread into the body cavities; via invasion of the lymphatic system through invasion of lymphatic cells and transport to regional and distant lymph nodes and then to other parts of the body; via the cardiovascular system; or via invasion of the surrounding tissue. Migrating cancers include metastatic tumors and cell migration cancers, such as ovarian cancer, mesothelioma, and primary lung cancer, each of which is characterized by cellular migration.

[0044] As used herein, “slowing the spread of migrating cancer” refers to reducing or stopping the formation of new loci; or reducing, stopping, or reversing the tumor load.

[0045] As used herein,“metastatic tumor” refers to a tumor or cancer in which the cancer cells forming the tumor have a high potential to or have begun to, metastasize, or spread from one location to another location or locations within a subject, via the lymphatic system or the cardiovascular system, for example, creating secondary tumors within the subject. Such metastatic behavior may be indicative of malignant tumors. In some cases, metastatic behavior may be associated with an increase in cell migration and/or invasion behavior of the tumor cells.

[0046] As used herein,“slowing the spread of metastasis” refers to reducing or stopping the formation of new loci, including stopping micrometastatic disease; or reducing, stopping, or reversing the tumor load.

[0047] The term“cancer” refers to any cancer caused by the proliferation of malignant neoplastic cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias, lymphomas, and the like.

[0048] Examples of cancers that can be defined as metastatic include but are not limited to non-small cell lung cancer, breast cancer, ovarian cancer, colorectal cancer, biliary tract cancer, bladder cancer, brain cancer including glioblastomas and medullablastomas, cervical cancer, choriocarcinoma, endometrial cancer, esophageal cancer, gastric cancer, hematological neoplasms, multiple myeloma, leukemia, intraepithelial neoplasms, livercancer, lymphomas, neuroblastomas, oral cancer, pancreatic cancer, prostate cancer, sarcoma, skin cancer including melanoma, eye cancers including ocular melanoma, basocellular cancer, squamous cell cancer, testicular cancer, stromal tumors, germ cell tumors, thyroid cancer, and renal cancer. [0049] “Proliferation” as used in this application involves reproduction or multiplication of similar forms (cells) due to constituting (cellular) elements.

[0050] “Cell migration” as used in this application involves the invasion by the cancer cells into the surrounding tissue and the crossing of the vessel wall to exit the vasculature in distal organs of the cancer cell.

[0051] By“cell migration cancers” is meant cancers that migrate by invasion by the cancer cells into the surrounding tissue or fluid, including those that migrate by way of blood vessels or cardiovascular tissues or fluids and may further exit the vasculature in distal organs.

[0052] “Non-metastatic cell migration cancer” as used herein refers to cancers that do not migrate via the lymphatic system, through the interstitium, or through the cardiovascular system.

[0053] “Administering” means providing a pharmaceutical agent or composition to a subject, and includes, but is not limited to, administering by a medical professional and self-administering.

[0054] “Parenteral administration,” means administration through injection or infusion. Parenteral administration includes, but is not limited to, subcutaneous administration, intravenous administration, intramuscular administration, intraarterial administration, intracranial administration, or administration to a nerve or nerve sheath.

[0055] “Subcutaneous administration” means administration just below the skin.

[0056] “Intravenous administration” means administration into a vein.

[0057] “Intraarterial administration” means administration into an artery.

[0058] “Ophthalmic administration” means administration to or across the surface of the eye or to the orbital space.

[0059] “Intraocular administration” means administration into the interior of the eye, such as into the vitreous body, or within the uveal, ciliary, corneal, macular, retinal, sub- retinal, or suprachoroidal spaces.

[0060] The term“agent” includes any substance, molecule, element, compound, entity, or a combination thereof. It includes, but is not limited to, e.g., protein, polypeptide, peptide or mimetic, small organic molecule, polysaccharide, polynucleotide, and the like. It can be a natural product, a synthetic compound, or a chemical compound, or a combination of two or more substances.

[0061] “Pharmaceutical agent” means a substance that provides a therapeutic effect when administered to a subject.

[0062] “Pharmaceutical composition” means a mixture of substances suitable for administering to an individual that includes a pharmaceutical agent. For example, a pharmaceutical composition may comprise a modified oligonucleotide and a sterile aqueous solution.

[0063] “Active pharmaceutical ingredient” means the substance in a pharmaceutical composition that provides a desired effect.

[0064] The term “pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” includes any and all solvents, diluents, emulsifiers, binders, buffers, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like, or any other such compound as is known by those of skill in the art to be useful in preparing pharmaceutical formulations. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions. In addition, various adjuvants such as are commonly used in the art may be included. These and other such compounds are described in the literature, e.g., in the Merck Index, Merck & Company, Rahway, NJ. Considerations for the inclusion of various components in pharmaceutical compositions are described, e.g., in Gilman et al. (Eds.) (1990); Goodman and Gilman’s: The Pharmacological Basis of Therapeutics, 8th Ed., Pergamon Press.

[0065] The embodiments as described herein may further be understood with reference to the following non-limiting examples.

Example 1

[0066] Primary cell lines derived from ocular melanomas are known in the art. See Stei, M. S. et ah, BioMed Res. Int. (2016), Article I.D. No. 4521807, doi:l0.1155/2016/4521807, which is hereby incorporated by reference in its entirety and with respect to its disclosure of ocular melanoma cell lines and in vivo and ex vivo models of OM. Primary OM cell lines and normal ocular melanocytes are cultured in the presence and absence of ranpirnase, with the resulting changes in cell viability measured using spectrophotometric measurements of yellow tetrazolium MTT (3-(4, 5-dimethylthiazolyl-2)- 2, 5-diphenyltetrazolium bromide) or an equivalent indicator of cellular metabolism and viability added and assayed according to manufacturer’s protocols. See e.g.,“MTT Cell Proliferation Assay Instruction Guide,” ATCC 30-1010K, American Type Culture Collection (2011) which is hereby incorporated by reference in its entirety. Results are evaluated for appropriate statistical measures of validity. Samples exposed to ranpirnase show reduced viability compared to samples not exposed to ranpirnase. OM cells show reduced viability compared to normal ocular melanocytes.

Example 2

[0067] Ranpirnase is administered to cancer-free mice by subchoroidal injection. Urine and feces are collected from each animal to be assayed for the presence of ranpirnase and/or metabolites thereof. If necessary, isotopically labeled ranpirnase can be used to facilitate measurement. Animals are sacrificed at appropriate intervals, and the amount of ranpirnase in various tissues, especially including the liver, eye, and blood are measured. Pharmacokinetic parameters including absorption, distribution, metabolism, and excretion are calculated.

Example 3

[0068] Syngeneic OM mice are generated by intraocular injection of cutaneous melanoma cells as described in Stei et al. and references therein. Ranpirnase is administered to the resulting mice by subchoroidal injection. Urine and feces are collected from each animal to be assayed for the presence of ranpirnase and/or metabolites thereof. If necessary, isotopically labeled ranpirnase can be used to facilitate measurement. Animals are sacrificed at appropriate intervals, and the amount of ranpirnase in various tissues, especially including the liver, eye, and blood are measured. Pharmacokinetic parameters including absorption, distribution, metabolism, and excretion are calculated. Eyes are dissected, stained, and imaged for the presence of OM cells. The extent and severity of experimental tumors is determined.

Example 4

[0069] Syngeneic OM mice are generated by intrahepatic injection of cutaneous melanoma cells as described in Stei et al. and references therein. Ranpirnase is administered to the resulting mice by intravenous and/or intrahepatic infusion. Urine and feces are collected from each animal to be assayed for the presence of ranpirnase and/or metabolites thereof. If necessary, isotopically labeled ranpirnase can be used to facilitate measurement. Animals are sacrificed at appropriate intervals, and the amount of ranpirnase in various tissues, especially including the liver, eye, and blood are measured. Pharmacokinetic parameters including absorption, distribution, metabolism, and excretion are calculated. Livers are dissected, stained, and imaged for the presence of OM cells. The extent and severity of experimental liver tumors is determined.

Example 5

[0070] Human xenograft OM mice are generated by intraocular injection of human OM cells as described in Stei et al. and references therein. Appropriate immounosuppressive agents are administered sufficient to allow tumor growth. Ranpirnase is administered to the resulting mice by subchoroidal injection. Urine and feces are collected from each animal to be assayed for the presence of ranpirnase and/or metabolites thereof. If necessary, isotopically labeled ranpirnase can be used to facilitate measurement. Animals are sacrificed at appropriate intervals, and the amount of ranpirnase in various tissues, especially including the liver, eye, and blood are measured. Pharmacokinetic parameters including absorption, distribution, metabolism, and excretion are calculated. Eyes are dissected, stained, and imaged for the presence of OM cells. The extent and severity of experimental eye tumors is determined. Example 6

[0071] Human xenograft OM mice are generated by intrahepatic injection of human OM cells as described in Stei et al. and references therein. Appropriate immounosuppressive agents are administered sufficient to allow tumor growth. Ranpimase is administered to the resulting mice by intravenous and/or intrahepatic infusion. Urine and feces are collected from each animal to be assayed for the presence of ranpirnase and/or metabolites thereof. If necessary, isotopically labeled ranpirnase can be used to facilitate measurement. Animals are sacrificed at appropriate intervals, and the amount of ranpirnase in various tissues, especially including the liver, eye, and blood are measured. Pharmacokinetic parameters including absorption, distribution, metabolism, and excretion are calculated. Livers are dissected, stained, and imaged for the presence of OM cells. The extent and severity of experimental liver tumors is determined.

Example 7

[0072] A subject suffering from ocular melanoma is identified, and a ranpimase composition as described herein is administered to the identified subject. The subject may be identified by eye examination, wherein the exterior of the eye is examined or by the use of slit-lamp microscopy to exam the interior of the eye. The subject may also or alternatively be identified by ultrasound, angiogram, optical coherence tomography, or by biopsy.

[0073] Upon identification, a composition comprising ranpimase as described herein is administered to the eye. The composition may be administered topically to the eye (such as in the form of a gel or eye drops), or by injection, such as by intravitreal injection.

Example 8

[0074] The experiments described in Examples 2-5 are carried out in the presence of additional immuno therapeutic agents, chemotherapeutic agents, checkpoint inhibitors, and/or other anticancer agents. Enhancements of the efficacy of ranpimase and/or synergistic effects are noted.

[0075] With respect to the use of substantially any plural and/or singular terms herein, those having skill in the art can translate from the plural to the singular and/or from the singular to plural as is appropriate to the context and/or application. The various singular/plural permutations can be expressly set forth herein for sake of clarity.

[0076] It will be understood by those within the art that, in general, terms used herein, and especially in the appended claims (for example, bodies of the appended claims) are generally intended as “open” terms (for example, the term “including” should be interpreted as“including but not limited to,” the term“having” should be interpreted as “having at least,” the term“includes” should be interpreted as“includes but is not limited to,” etc.). It will be further understood by those within the art that if a specific number of an introduced claim recitation is intended, such an intent will be explicitly recited in the claim, and in the absence of such recitation no such intent is present. For example, as an aid to understanding, the following appended claims can contain usage of the introductory phrases “at least one” and“one or more” to introduce claim recitations. However, the use of such phrases should not be construed to imply that the introduction of a claim recitation by the indefinite articles“a” or“an” limits any particular claim containing such introduced claim recitation to embodiments containing only one such recitation, even when the same claim includes the introductory phrases“one or more” or“at least one” and indefinite articles such as“a” or“an” (for example,“a” and/or“an” should be interpreted to mean“at least one” or “one or more”); the same holds true for the use of definite articles used to introduce claim recitations. In addition, even if a specific number of an introduced claim recitation is explicitly recited, those skilled in the art will recognize that such recitation should be interpreted to mean at least the recited number (for example, the bare recitation of “two recitations,” without other modifiers, means at least two recitations, or two or more recitations). Furthermore, in those instances where a convention analogous to“at least one of A, B, and C, etc.” is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (for example,“ a system having at least one of A, B, and C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and/or A, B, and C together, etc.). In those instances where a convention analogous to“at least one of A, B, or C, etc.” is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (for example,“ a system having at least one of A, B, or C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and/or A, B, and C together, etc.). It will be further understood by those within the art that virtually any disjunctive word and/or phrase presenting two or more alternative terms, whether in the description, claims, or drawings, should be understood to contemplate the possibilities of including one of the terms, either of the terms, or both terms. For example, the phrase“A or B” will be understood to include the possibilities of“A” or“B” or“A and B.”

[0077] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0078] As will be understood by one skilled in the art, for any and all purposes, such as in terms of providing a written description, all ranges disclosed herein also encompass any and all possible sub-ranges and combinations of sub-ranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as“up to,”“at least,”“greater than,”“less than,” and the like include the number recited and refer to ranges which can be subsequently broken down into sub-ranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 articles refers to groups having 1, 2, or 3 articles. Similarly, a group having 1-5 articles refers to groups having 1, 2, 3, 4, or 5 articles, and so forth.

[0079] While various aspects and embodiments have been disclosed herein, other aspects and embodiments will be apparent to those skilled in the art. The various aspects and embodiments disclosed herein are for purposes of illustration and are not intended to be limiting, with the true scope and spirit being indicated by the following claims.