COMPOSITION AND A COPPER COMPOSITION FOR TREATMENT OF DERMA TOLOGIC CONDITIONS

RELATED APPLICATIONS

[0001] This application claims the benefit under 35 U.S.C. § 119(e) as a nonprovisional application of U.S. Prov. App. No. 61/909,281 filed on November 26, 2013, which is hereby incorporated by reference in its entirety,

BACKGROUND

Field of the invention

[0002] Embodiments of the invention relate to delivery of hyaluronic acid (HA) compositions into the skin, and methods of increasing the half-life thereof.

Description of the Related Art

[0003] Hyaluronic acid has been used for a wide variety of medical applications, including dermatologic applications. Hyaluronic acid is degraded in vivo by hyaluronidases, thus requiring frequent periodic re-dosing. What is needed are systems and methods to increase the half-life of the HA composition in order to advantageously prolong the therapeutic effects and d ecrease the frequency of re-dosing.

SUMMARY

[0004] Disclosed herein is a method of treating a dermatologic condition, comprising the steps of: delivering a hyaluronic acid composition into the skin; and administering a copper-containing skin formulation prior to, during, and/or following delivery of the HA composition. In some embodiments, the HA composition is iontophoretically delivered. In some embodiments, the copper-containing skin formulation is topically delivered. The HA composition can comprises palmitoyl-KTTK. The copper- containing skin formulation comprises copper tripeptide-l . The copper-containing skin formulation can be topically administered within, for example, within about 6 hours, within about 3 hours, or within about 1 hour of the delivering of the HA composition. In some embodiments, topically administering a copper-containing skin formulation increases the half- life of the HA composition within the skin by at least about 10%, 25%, or more. The dermatologic condition could include, for example, wrinkles. Not to be limited by theory, but copper-containing compositions can unexpectedly and synergistically inhibit hyaluronidases and significantly increase the efficacy and/or half-life of HA compositions when coadministered or both administered in a relatively short temporal period with respect to each other.

[0005] The methods summarized above and set forth in further detail below describe certain actions taken by a practitioner; however, it should be understood that they can also include the instruction of those actions by another party. Thus, actions such as "administering a copper composition" include "instructing the administration of a copper composition,"

BRIEF DESCRIPTION OF THE DRAWINGS

[0006] FIG. 1 illustrates an embodiment of a method of treating a dermatological condition including the steps of delivering a H A formulation info the skin, and delivering a copper formulation into the skin.

DETAILED DESCRIPTION

Hyaluronic Acid

[0007] Hyaluronan (also called hyaluronic acid or hyaluronate or HA) is an anionic, nonsulfated glycosaminoglycan distributed widely throughout connective, epithelial, and neural tissues. It is unique among glycosaminoglycans in that it is nonsulfated, forms in the plasma membrane instead of the Golgi, and can be very large, with its molecular weight often reaching the millions. One of the chief components of the extracellular matrix, hyaluronan contributes significantly to cell proliferation and migration. Hyaluronan is also a major component of skin, where it is involved in tissue repair. When skin is exposed to excessive UYB rays, it becomes inflamed and the cells in the dermis stop producing as much hyaluronan, and increase the rate of its degradation. Hyaluronic acid can be degraded in the skin enzymatically, such as via hyaluronidases. Hyaluronan degradation products then accumulate in the skin after UV exposure.

[0008] High molecular weight HA as used herein describes a HA material having a molecular weight of at least about 1.0 million Daltons (mw≤_106 Da or 1 MDa) to about 4.0 MDa. For example, the high molecular weight HA in the present compositions may have a molecular weight of about 2.0 MDa. In another example, the high molecular weight HA may have a molecular weight of about 2.8 MDa.

[0009] Low molecular weight HA as used herein describes a HA material having a molecular weight of less than about 5.0 MDa. Low molecular weight HA can have a molecular weight of between about 200,000 Da (0.2 MDa.) to less than about 1.0 MDa, for example, between about 300,000 Da (0.3 MDa) to about 750,000 Da. (0.75 MDa). In some embodiments, the HA has a molecular weight of between about 750kDa and about 950kDa, such as about 85()kDa; between about 20kDa and about 5()kDa, such as about 30kDa; or between about 5kDa and about 20 kDa, such as about 7kDa; or between about 5kDa and about lOOkDa. In some embodiments, the hyaluronic acid component encompasses a range of hyaluronic acids having a distribution of molecular weights, such as a Gaussian distribution in some cases. As such, the molecular weight can be expressed as an average molecular weight reflecting a varying distribution of hyaluronic acid species having different molecular weights. In some embodiments, the HA can be a sodium hyaiuronate and can have a molecular weight or an average molecular weight of between about 250,000 Da and about 450,000 Da, such as between about 300,000 Da and about 400,000 Da, or about 300,000 Da, 310,000 Da, 320,000 Da, 330,000 Da, 340,000 Da, 350,000 Da, 360,000 Da, 370,000 Da, 380,000 Da, 390,000 Da, or 400,000 Da. In some embodiments, the HA can have a molecular weight or an average molecular weight of between about 0.85 M Da and about 3 MDa, between about 0.85 MDa and about 1.6 mDa, between about 1.6mDa and about 2.9 MDa, or about 0.85, 0.90, 0.95, 1.00, 1.05, 1.1, 1.15, 1.2, 1.25, 1.30, 1.35, 1.40, 1.45, 1.50, 1.55, 1.60, 1.65, 1.70, 1.75, 1.80, 1.85, 1.90, 1.95, 2.00, 2.05, 2.10, 2.15, 2.20, 2.25, 2,30, 2.35, 2.40, 2,45, 2.50, 2.55, 2.60, 2.65, 2.70, 2,75, 2.80, 2.85, 2.90, 2.95, 3.00 mDa, or any range including two of the previous values.

10010] in some embodiments, the low molecular weight HA can make up between about 0.5% and about 50% w/w percent of the composition, such as between about 1% and about 30% w/w, between about 5% and about 30% w/w, between about 10% and about 25% w/w, between about 10% and about 20% w/w, or about 8%, 9%, 10%, 1 1%, 12%, 13%», 14%, 15%, 16%», 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30% w/w of the composition, or a range including any two of the preceding values. [ΘΘ11] In some embodiments, the high molecular weight HA can make up between about 0.5% and about 10% w/w percent of the composition, such as between about 0.5% and about 3% w/w, between about 1 %> and about 3% w/w, between about 1% and about 2% w/w, or about 0.5%, 0.75%, 1%, 1.25%, 1 .5%, 1.75%, 2%, 2.25%, 2.5%, 2.75%, 3%, 3.5%, 4%, 4.5%, 5% w/w of the composition, or a range including any two of the preceding values,

[ΘΘ12] In some embodiments, the composition could also include one or more chitosans that could include, for example, ultrapure chitosan salts and bases. Some suitable chitosans are Protasans from NovaMatrix; Sandvika., Norway. PROTASAN UP CL 1 13 is based on a chitosan where between 75-90 percent of the acetyl groups are removed. The cationic polymer is a highly purified and well-characterized water-soluble chloride salt. The functional properties are described by the molecular weight and the degree of deacetylation. Typically, the molecular weight for PROTASAN UP CL 1 13 is in the 50000-150000 g/mol range (measured as a chitosan acetate). PROTASAN UP CL 1 14 is based on a chitosan where more than 90 percent of the acetyl groups are removed. The cationic polymer is a highly purified and well-characterized water-soluble chloride salt. The functional properties are described by the molecular weight and the degree of deacetylation. Typically, the molecular weight for PROTASAN UP CL 1 14 is in the 50000— 150000 g/mol range (measured as a chitosan acetate). PROTASAN UP CL 21 3 is based on a chitosan where between 75-90 percent of the acetyl groups are removed. The cationic polymer is a highly purified and well-characterized water-soluble chloride salt. The functional properties are described by the molecular weight and the degree of deacetylation. Typically, the molecular weight for PROTASAN UP CL 213 is in the 150000-400000 g mol range (measured as a chitosan acetate), PROTASAN UP CL 214 is based on a chitosan where more than 90 percent of the acetyl groups are removed. The cationic polymer is a highly purified and well- characterized water-soluble chloride salt. The functional properties are described by the molecular weight and the degree of deacetylation. Typically, the molecular weight for PROTASAN UP CL 214 is in the 150000-400000 g/mol range (measured as a chitosan acetate). PROTASAN UP G 1 13 is based on a chitosan where between 75-90 percent of the acetyl groups are removed. The cationic polymer is a highly purified and well-characterized water-soluble chloride salt. The functional properties are described by the molecular weight. and the degree of deacetylation. Typically, the molecular weight for PROTASAN UP G 1 13 is in the 50000-150000 g/mol range (measured as a chitosan acetate). PROTASAN UP G 1 14 is based on a chitosan where more than 90 percent of the acetyl groups are removed. The cationic polymer is a highly purified and well-characterized water-soluble chloride salt. The functional properties are described by the molecular weight and the degree of deacetylation. Typically, the molecular weight for PROTASAN UP G 1 14 is in the 50000—150000 g/mol range (measured as a chitosan acetate). PROTASAN UP G 213 is based on a chitosan where between 75-90 percent of the acetyl groups are removed. The cationic polymer is a highly purified and well-characterized water-soluble chitosan glutamate. The functional properties are described by the molecular weight, and the degree of deacetylation. Typically, the molecular weight for PROTASAN UP G 213 is in the 150000-600000 g/mol range (measured as a chitosan acetate). PROTASAN UP G 214 is based on a chitosan where more than 90 percent, of the acetyl groups are removed. The cationic polymer is a highly purified and well-characterized water-soluble chloride salt. The functional properties are described by the molecular weight and the degree of deacetylation. Typically, the molecular weight for PROTASAN UP G 214 is in the 150000-400000 g/mo! range (measured as a chitosan acetate). In some embodiments, the chitosan can make up between about 0.5% and about, 50% w/w percent of the composition, such as between about 1% and about 25% w/w, between about 1% and about 10% w/w, between about 1% and about 5% w/w, , between about 2% and about 3% w w, or about 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% w/w of the composition.

[00Ϊ3] Degree of crosslinking as used herein refers to the intermoiecular junctions joining the individual HA polymer molecules, or monomer chains, into a permanent structure, or as disclosed herein the soft tissue filler composition. Moreover, degree of crosshnking for purposes of the present disclosure is further defined as the percent weight ratio of the crosshnking agent to HA-monomeric units within the crosslinked portion of the HA based composition. t is measured by the weight ratio of HA monomers to crosslinker (HA monomers rosslinker).

[0014] Free HA as used herein refers to individual HA polymer molecules that are not crosslinked to, or very lightly crosslinked to (very low degree of crosslinking) the highly crosslinked (higher degree of crosslinking) maeromoleeular structure making up the soft tissue filler composition. Free HA generally remains water soluble. Free FIA can alternatively be defined as the "uncrosslinked," or lightly erosslinked component of the macromolecular structure making up the soft tissue filler composition disclosed herein.

[0015] Cohesive as used herein is the ability of a FLA -based composition to retain its shape and resist deformation. Cohesiveness is affected by, among other factors, the molecular weight ratio of the initial free HA, the degree of crosslinking, the amount of residual free HA following crosslinking, and HA-based composition pH.

[0016] As used herein, hyaluronic acid (HA) can refer to any of its hyaluronate salts, and includes, but is not limited to, sodium hyaluronate (NaHA), potassium hyaluronate, magnesium hyaluronate, calcium hyaluronate, and combinations thereof.

[0017] In some embodiments, the concentration of HA in the compositions described herein is preferably at least 10 mg/mL and up to about 40 mg/mL, For example, the concentration of HA in some of the compositions is in a range between about 20 mg/mL and about 30 mg/mL. In some embodiments, the HA comprises between about 0.1 % and about 15% by weight of the entire composition, between about 0.5% and about 5% by weight of the entire composition, between about 1% and about 5% by weight of the entire composition; or about 1%, 2%, 3%, 4%, or 5% by weight of the entire composition.

[0018] In some embodiments of the present invention, the HA component of the present compositions, hereinafter sometimes, "precursor composition" is a hydrated, cohesive gel. A cohesive gel, relative to a non-cohesive gel, is better able retain its shape and resist deformation, for example, after being subjected to shear or other stresses. In some cases, such cohesive gels can be less likely to substantially degrade or become unstable over time or when subjected to external stimuli such as sterilization, relative to non-cohesive gels. However, both cohesive and non-cohesive gels can be used in certain embodiments.

[0019] In some embodiments, HA can be eornp!exed with a suitable crosslinking agent. The crosslinking agent may be any agent known to be suitable for crosslinking polysaccharides and their derivatives via their hydroxyl groups. Suitable crosslinking agents include, but are not limited to, 1 ,4-butanediol diglycidyl ether (or l ,4-bis(2,3- epoxypropoxyjbutane or 1 ,4-bisglycidyloxybutane, all of which are commonly known as BDDE), 1 ,2-bis(2,3-epoxypropoxy)ethylene and l-{2,3-epoxypropyI)-2,3-epoxyeyelohexane. The use of more than one crosslinking agent or a different crosslinking agent is not excluded from the scope of the present disclosure. The step of crosslinking may be carried out using any means known to those of ordinary skill in the art. Those skilled in the art appreciate how to optimize conditions of crosslinking according to the nature of the HA, and how to carry out crosslinking to an optimized degree. Degree of crosslinking for purposes of the present disclosure is defined as the percent weight ratio of the crosslinking agent to HA-monomeric units within the crosslinked portion of the HA based composition. It is measured by the weight ratio of HA monomers to crosslinker (HA monomers:crosslinker). In some embodiments, the degree of crosslinking in the HA component of the present compositions is at least about 2% and is up to about 20%, In other embodiments, the degree of crosslinking is greater than 5%, for example, is about 6% to about 8%. In some embodiments, the degree of crosslinking is between about 4% to about 12%. In some embodiments, the degree of crosslinking is less than about 6%, for example, is less than about 5%. In some embodiments, the HA component is capable of absorbing at least, about one time its weight in water. When neutralized and swollen, the crosslinked HA component and water absorbed by the crosslinked HA component is in a weight ratio of about 1 : 1. Some embodiments of resulting hydrated HA-based gels have a characteristic of being highly cohesive.

Amphophilic Peptide Component

[ΘΘ20] In some embodiments, the HA composition also includes an amphophilic peptide, e.g., a palmitoyl compound. Some non-limiting examples of palmitoyl compounds include palmitoyl TTK, palmitoyl TTKS, palmitoyl GH , and palmitoyl hexapeptide, among others. Not to be limited by theory, it is believed that palmitoyl groups in particular enhance the hydrophobic! ty and contribute to membrane association, similar to S- Palmitoylation observed with proteins. The association of the fatty acid chain is reversible (because the bond between palmitic acid and glutathione is a thio-ester bond) allowing the compound to be absorbed by the cell membranes. In combination with HA, palmitoyl compounds may synergistically increase penetration of the HA into a desired region, such as the dermis layer of the skin, by reducing the negative charge of the HA. In some embodiments, the composition can include between about 0-2%, such as less than about 1%, 0.5%, 0.3%, 0.2%, 0.1%, 0.09%, 0.07%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, or 0.01% of an amphophilic peptide. Delivery into an Anatomical Targe Region

[0021] The HA composition can be delivered to a target region in the body by any desired route. In some embodiments, the HA composition can be delivered to the target region topically, subcutaneously, intramuscularly, or intraarticularly, for example. In some embodiments, the HA composition can be delivered iontophoreticaliy/electrostatically. lontophoretic/electrostatic systems and methods that can involve a HA composition are described, for example, in U.S. Pat. Pub. No. 2011/0190724 Al to Francis et al., hereby incorporated by reference in its entirety as well as attached hereto as Appendix A.

[0022] The HA composition can be delivered to the target region of the skin at any desired frequency depending on the desired clinical result. In some embodiments, the HA. composition can be delivered daily, every other day, every 3 days, 3 times a week, 2 times a week, once weekly, every 10 days, every 14 days, every month, every quarter, or at other specified intervals.

Copper Formulations

[0023] Not to be limited by theory, copper formulations can synergistically inhibit hyaluronidase, as well as have an anti-inflammatory effect on the skin. Certain copper formulations can be positively charged, and as such comple with HA compounds and retain them within a desired anatomical region, such as the dermis of the skin. Such formulations, applied topically for example, in conjunction with delivery of an HA compound in the skin, can advantageously to inhibit HA degradation and increase half-life of the HA. In some embodiments, the copper formulation can be applied subcutaneously, intramuscularly, or intraarticularly, for example. In some embodiments, the copper composition can be delivered iontophoreticaUy/electrostatically, together with or separate from the HA formulation, in some embodiments, a copper formulation is delivered into a joint along with a HA composition, to reduce the frequency of injection required to treat a condition such as osteoarthritis, rheumatoid arthritis, or other arthridities and arthropathies, for example.

[0024] In some cases, copper salts alone can be ineffective, or even inhibitory, for such applications. The copper can be delivered in a biologically acceptable form. As an example, when copper is complexed with a biologically acceptable carrier molecule, such as a peptide, it may then be effectively delivered to cells. More specifically, peptide copper complexes that have utility for wound healing and skin health when topically applied are described in U.S. Patent Nos. 4,760,051 ; 4,665,054; 4,877,770; 5, 135,913 and 5,348,943, each of which are incorporated by reference in their entireties. As used herein, the term "peptide copper complex" refers to a coordination compound comprising a peptide molecule and a copper ion non~ eovalently complexed therewith. The peptide molecule serves as the complexing agent by donating electrons to the copper ion to yield the non- covaient complex. The peptide molecule is a chain of two or more amino acid units eovalently bonded together via amide linkages (for example, -CONH-), the formation of such linkages being accompanied by the elimination of water. The amino acid units are from amino acids that are naturally occurring or ofhenvise. Also, at least one amide linkage nitrogen atom may have eovalently bonded thereto either a hydrogen atom or another moiety.

[0025] One example of a copper peptide complex is alanyl-hi.stid.yl- lysine :copper(ll). Copper(II), as is well understood by the skilled artisan, designates a copper ion having a valence of 2 (e.g., Cix' ⁺ ). Additional examples of the peptide copper complexes, encompassed in embodiments of the present, invention, include, but are not limited to, those described in U.S. Patent Nos. 4,665,054; 4,760,051 ; 4,767,753; 4,877,770; 5,023,237; 5,059,588; 5,120,831 ; 5,135,913; 5,145,838; 5, 177,061 ; 5,214,032; 5,348,943; 5,538,945 5,550,183, and U.S. Pat. Pub. No. 2003/0134780 Al , each of which are incorporated herein by reference in their entireties. In certain specific embodiments, the composition of the present invention comprises at least one peptide copper complex that is alanyi-histidyl- lysine:copper(Il) ("AH -Cu"), valyl-histidyl-lysine:copper(n) ("VH -Cu"), or glycyl- histidyl-lysine:copper(II) ("GHK-Cu"), Such peptides may be in either the L or D form, in a related, more specific embodiment, they are all in the L form . Further, the expression "peptide copper complex," as used herein, encompasses peptide copper complex derivatives. The expression "peptide copper complex derivative," as used herein, refers to a peptide copper compl ex where the peptide molecule thereof has: 1 ) at least one amino acid side chain moiety that is a modification and/or variation of a naturally occurring, amino acid side-chain moiety; and/or 2) at least one of the hydrogens, bonded to an amide linkage nitrogen atom, substituted with a different moiety; and/or 3) the carboxyl group of the carboxyl terminal residue esterified or otherwise modified; and/or 4) at least one hydrogen, bonded to the nitrogen atom of the amino-terminal residue, substituted with a different moiety.

[0026] The amino acid side-chain moieties of the peptide copper complex derivatives may include alkyl, aryl, arylalkyl, alkoxy, or aryloxy moieties. As used herein, "aikyl" means a straight chain or branched, cyclic or noncyclic, substituted or unsubstituted, saturated or unsaturated aliphatic hydrocarbon containing from 1 to 18 carbon atoms. Representative saturated straight chain alkyls include methyl, ethyl, n-propyl and the like; while saturated branched alkyls include isopropyl, sec-butyl, isohutyl, fetf-butyl, isopentyl, and the like. Representative, saturated cyclic alkyls include cyclopropyl, cyclobutyl, cyclopentyl, -CHbcyclohexyl, and the like; while unsaturated cyclic alkyls include cyclopentenyl, cyclohexenyl, and the like. Unsaturated alkyls contain at least one double or triple bond between adjacent carbon atoms (referred to as an "alkenyl" or "aikynyl," respectively). Representative alkenyls include ethylenyl, 1-butenyl, isobutylenyl, 2-methyl-2- butenyl, and the like; while representative alkynyls include acetylenyl, 2-butynyl, 3-methyl- 1-butynyl, and the like.

[0027] Also, as used herein, "aryl" means an aromatic carbocyclic moiety such as phenyl or naphthyi, and may be substituted or unsubstituted. "Arylalkyl," as used herein, means an aikyl having at least one alkyl hydrogen atom replaced with a substituted or unsubstituted aryl moiety, such as benzyl (i.e., -CH ₂ pbenyl, -(CH ₂ ) ₂ phenyl, -(C!¾) ₃ phenyl, - CH(phenyi) ₂ , and the like). As some examples, the amino acid side-chain moieties of alanine, valine, leucine, isoleucine and phenylalanine may generally be classified as alkyl, aryl or arylalkyl moieties. "Alkoxy" and "aryloxy," as used herein, refer, respectively, to alky and aryl moieties, as defined above, but each further comprising an oxygen atom used to link the moiety to the amino acid.

[0028] Additionally, the peptide copper complex derivative may, for example, be N-alkylated at one or more peptide bonds; and/or its carboxyl terminus may be esterified, for example, with a methyl, ethyl, or benzyl group, or may be reduced to a. hydroxy or aldehyde. Additionally, the peptide copper complex derivative may, for example, be N -alkylated, N- acylated or N- suifonylated at the amino terminus with, for example, methyl, benzyl, acetyl, benzoyl, methanesulfonyl, or fluorenyloxycarbonyl moieties. [0029] Examples of the peptide copper complex derivatives, encompassed in embodiments of the present invention, include, but are not limited to, those disclosed and described in the above-cited U.S. Patents that are directed to peptide copper complexes. As one specific example, the disclosed composition may comprise a peptide copper complex derivative that is a derivative of GHK-Cu. having the general formula:

[ΘΘ30] [glycyl-histidyl-lysme-R] : copper(II)

[0031] where R is an alkyl moiety containing from 1 to 18 carbon atoms, an aryl moiety containing from 6 to 12 carbon atoms, an aryla!kyl moiety, an a!koxy moiety containing from 1 to 12 carbon atoms, or an aryloxy moiety containing from 6 to 12 carbon atoms. This derivative of GHK-Cu is further described in the above-cited U.S. Patents that are directed to peptide copper complexes.

[0032] The above compositions may be prepared from aqueous solutions of peptide copper complexes. For example, an amount of dried peptide copper complex suitable for a desired concentration is readily dissolved in water with mixing and gentle heating. An alternati ve method is to prepare a solution of the desired peptide, followed by the addition of a copper salt in the desired molar ratio to yield the desired solution of the peptide copper complex. Examples of copper salts that may be used are cupric chloride and cupric acetate. When aqueous solutions of peptide copper complexes are prepared, the solutions are neutralized, typically with NaOFI, In various embodiments, the concentration of the at least one peptide copper complex, by weight, of the composition, ranges from about 0.01% to about 5%, from about 0.025% to about 1%, and from about 0.05% to about 0.5%, respectively, or about 0.5% or about 1.0%. Also, the molar ratio of peptide to copper in the complex ranges from about 1 : 1 to about 3: 1 in some embodiments, and from about 1 : 1 to about 2: 1 in other embodiments. In some embodiments, compositions can be formed by combining at least one peptide copper complex with retinol, at least one retinol derivative, or a mixture thereof, where the combined compounds are encapsulated in liposomes or microsponges to aid in the delivery of the peptide copper comple or to increase the stability of the composition. In yet another embodiment of such compositions, the combined compounds may be formulated in an instrument allowing the delivery of the compounds via iontophoresis.

[0033] Certain copper compounds have been shown to penetrate the stratum corneum, such as a copper peptide, such as a copper tri-peptide and hexa-peptide, for example. In some embodiments, the copper peptide is GHK-Cu (Glycl-L-hidtidyl-L-lysine copper cuprate, diacetate GHK-Cu(Ac) ₂ . In some embodiments, the copper compound can be part of a sunscreen preparation.

Method and Timing of Delivery

[0034] The copper formulation can be delivered before, during, and/or after delivery of the HA formulation, in some embodiments, the copper formulation is delivered within about 2 hours, 10 hours, 8 hours, 6 hours, 4 hours, 3 hours, 2 hours, 1 hour, 45 minutes, 30 minutes, 15 minutes, or less relative (either before, after, or both) to the delivery of the HA formulation.

[0035] In some embodiments, the topically administering a copper-containing skin formulation increases the half-life of the HA composition within the skin by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, 100%, or more relative to the half-life of the HA composition without co-administering a copper-containing skin formulation before, after, or during administration of the HA composition.

[0036] in some embodiments, administering the HA composition and the copper composition occur separately. In some embodiments, the HA composition and the copper composition can be administered simultaneously, and even within a single combined formulation, for example.

Indications

[0037] The compositions help treat or prevent any number of conditions, including dermatologic conditions such as severe skin dryness, dullness, loss of elasticity, lack of radiance, exaggerated lines and wrinkles, spider vessels or red blotchiness. In some embodiments, "marionette" lines, smile lines, deep nasolabial fold lines, crow's feet, fine lines/wrinkles, vertical lines between the eyebrows, horizontal forehead lines, sagging thin/frail skin, skin redness and dullness may be improved using compositions as described herein. The compositions can also be used in the prevention and treatment of: photodamaged skin, the appearance of fine lines and wrinkles, age spots, and aged skin. The disclosed composition can also increasing the flexibility of the stratum corneum, increasing the content of collagen and/or glycosaminoglycans in skin, increasing moisture in skin, decreasing transcutaneous water loss, and generally increasing the quality of skin. The disclosed composition also provides topical formulations effective in promoting a healthy scalp, and thereby useful in the prevention of hair loss, and as a. treatment before and after hair transplant surgical procedures.

[0038] It is contemplated that various combinations or subcombinations of the specific features and aspects of the embodiments disclosed above may be made and still fall within one or more of the inventions. Further, the disclosure herein of any particular feature, aspect, method, property, characteristic, quality, attribute, element, or the like in connection with an embodiment can be used in all other embodiments set, forth herein. Accordingly, it should be understood that various features and aspects of the disclosed embodiments can be combined with or substituted for one another in order to form varying modes of the disclosed inventions. Thus, it is intended that the scope of the present inventions herein disclosed should not be limited by the particular disclosed embodiments described above. Moreover, while the invention is susceptible to various modifications, and alternative forms, specific examples thereof have been shown in the drawings and are herein described in detail. It should be understood, however, that the invention is not to be limited to the particular forms or methods disclosed, but to the contrary, the invention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the various embodiments described and the appended claims. Any methods disclosed herein need not be performed in the order recited. The methods disclosed herein include certain actions taken by a practitioner; however, they can also include any third-party instruction of those actions, either expressly or by implication. For example, actions such as "administering a copper formulation" include "instructing the administration of a copper formulation." The ranges disclosed herein also encompass any and all overlap, sub-ranges, and combinations thereof. Language such as "up to," "at least," "greater than," "less than," "between," and the like includes the number recited. Numbers preceded by a term such as "about" or "approximately" include the recited numbers. For example, "about, 3 mm" includes "3 mm."