US 20060293227A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2006/0293227 A1 Bhatnagar (43) Pub. Date: Dec. 28, 2006

(54) COSMETIC COMPOSITIONS AND Publication Classification METHODS USING TRANSFORMING GROWTH FACTOR-BETAMMICS (51) Int. Cl. A6II 38/08 (2006.01) (76) Inventor: Rajendra S. Bhatnagar, Burlingame, A6II 38/18 (2006.01) CA (US) (52) U.S. Cl...... 514/12: 514/16; 514/17 Correspondence Address: WILSON SONSN GOODRCH & ROSAT (57) ABSTRACT 650 PAGE MILL ROAD PALO ALTO, CA 94304-1050 (US) The invention provides methods, compositions, and kits for a variety of cosmetic uses, where the methods, compositions (21) Appl. No.: 11/166,259 and kits utilize Substances that display one or more activities of transforming growth factor-3 (TGF-B mimics). Compo (22) Filed: Jun. 24, 2005 sitions, methods, and kits contain TGF-B mimics. Patent Application Publication Dec. 28, 2006 Sheet 1 of 2 US 2006/0293227 A1 FIG. I (A)

1 H ACE 1 1.952 1.249 .316 2 CH3 ACE 1 1927 2.333 .203 3 H2 ACE 1 1.576 2.782 1.133 4 H3 ACE 1 1.243 2.593 - 605 5 C ACE 1 3.323 2.853 -.119 6 O ACE 4.272 2.077 s. 20 7 N ALA 2 3.439 4.166 a.310 8 HN ALA 2 2.632 4.767 - .211 . 9 CA ALA 2 4 695 4.857 -.543 10 HA ALA 2 5.425 4.521 .195 1. CB ALA 2 5 21 4.572 - .957 12 HB1 ALA 2 4.475 4.894 -2.694 13 HB2 ALA 2 6.142 5.115 -2.122 14 HB3 ALA 2 5.399 3.506 -2.083 15 C ALA 2 4.443 6.350 - .361 16 O ALA 2 3.292 6.788 - .383 17 N. ASN 3 5504 7.30 - 187 18 HN ASN 3 6.426 5.699 - 147 19 CA ASN 3 5.484 8.570 .015 20 HA ASN 3 A836 9,029 - 733 2 CB ASN 3 4.954, 887, 1,432 22 HB2 ASN 3 5.570 8.464. 2.158 23 HB3 ASN 3 4.000 8.383 1578 24 CG ASN 3 4.760 10.357 1.704 25 OD, ASN 3 4.723 11177 792 26 ND2: ASN 3 4.654 10.739 2.969 27 HND ASN 3 4.655 0.052 3709 28 HND2 ASN 3 4.487 11.714 3.168 29 C ASN 3 - 6925 9.054 - 180 30 O ASN 3 7.816 8.242 - .407 3 N WAL. 4 7.195 0.354 - 082 32 h WAL 4 6.433 1.007 .038 33 CA WA 4 8.563 10.863 - 10 34 HA VAL 4 9.001 10.579 -1.068 35 CB WAL. 4 8.546 12.404 - 038 36 HB WAL 4 7.922 2.763 - 857. 37 CG, WA 4 7,958 12.95 1272 38 HG WA 4 8:590 12684. 2.19 39 HG 2 VA 4 7900 14038 1213 40 HG13 VAL 4 6.955 12.563 1.438 A1 CG2 WAL 4, 9,949 2.988 - 245 42 HG21. WA 4 10.376 12.608 -1.174. 43 HG22 VAL 4 9.889 14.075 - .306 44 HG2.3 WAL. 4 10605 12.718 .583 45 C VAL 4 94-14 10.217 .994 46 0 WAL. 4 10604. 9.986 807 47 N AA 5 8,808 9.966 2.157 48 HN ALA 5 7.829 10.76 2.244 49 CA ALA 5 9.520 9482 3,331 50 HA ALA 5 10.41110.096 3.479 Patent Application Publication Dec. 28, 2006 Sheet 2 of 2 US 2006/0293227 A1 FIG. / (B)

5. CB ALA 8.626 9,645 4.563 52 HB ALA 7723 9,042 4.457 53 HB2 ALA 969 9.34 5.45 54 HB3 ALA 8,350 10.693 4,688 55 C ALA 9958 8025 3.185 55 O ALA 1,076 7.684 3.587 - 57 N G . . . 9,079 71.68 2.682 58 HN G 8.245 7,512 2,233 59 CA GL 9.204 5,720 225 50 A GLU. 0.254 5.429 2.779 61 CB GLU 8.456 5.20 3.964 62 HB2 GLU 7.393 5.439 3.867 63 HB3 GLU 8.837 5,712 4854 64. CG GL 8,602 42O3 65 HG2 GLU 9,655 3.443 4.33 55 HG3 GLU - 8.176 3.38 3.375 67 786. 3290 54.67 58 6,706 3.745 5.612 59 8.468 2,548 6.266 . 70 8.583 588 1438 71 . 7533 5,781 930 72 923 4,107 882 73 9.827. 3.588 1392 74 8,673 3,542 a .382 75 7,595 3.576 .478 76 9,39) 4.236 -1.545 77 10.466 4.108 -1.419 78 9.155 5.30 -530 79 8,952 3.664 2.892 80 7.842 397 -3055 81. 9,840 3699 -3.887 82 HND ASN 0.757 4.091 -3 745 .83 HNO2 ASN 9.556 3.315 -4 774 84 C ASN 8974 2,049 - .371 85 O ASN 9950 1634 253 , 86 N ALA 8.148 1.245 1.035 87 N ALA 7.449 1658 -6A4 88 CA ALA 8.215 -205 -1.003 89 HA ALA 9.244 -529 -837 90 CB ALA 7.341 -77 150 9. HB ALA 6,300 -440 - 024 92 HB2 ALA 7,415 - 800 241. 93 HB3 ALA 7675 -269 1087 94. C ALA 7.762 -755 -2.355 95 O AA 7.562 -002 -3.304 95 N. NME 7.62 -2O77 -2,453 97 HN NME 7.783 -2.641 -1.638 98 CT NME 755 -2736 -3573 99 HT NME 6,43 -2.126 -4 2O1 OO HT2 NME 8,026 -2,904 4.322 10 HT3 NME 6,712 -3.695 -3 424 US 2006/0293227 A1 Dec. 28, 2006

COSMETC COMPOSITIONS AND METHODS about once per day to about five times per day. In some USING TRANSFORMING GROWTH embodiments, the TGF-3 mimic is administered at an aver FACTOR-BETAMMICS age frequency of about once per day to about once per week. In some embodiments, the TGF-B mimic is administered at BACKGROUND OF THE INVENTION an average frequency of less than about once per day. 0001 Skin is subject to a number of conditions that can 0005. In embodiments of methods of the invention, the lead to a desire for cosmetic enhancement. Such conditions TGF-B mimic is applied in a vehicle Such as, e.g., a spray, include aging due to chronological aging or photoaging from ointment, gel, lotion, milk, liposomal preparation, or patch. exposure to the Sun, or both. Skin aging results in wrinkling, In some embodiments, the vehicle is a lotion. In some the appearance of pigmented areas (“age spots”), thinning of embodiments of the invention, the lotion includes a mixture the skin, loss of elasticity, and other undesirable character of emulsifying lanolin alcohols, waxes, and oils. In some istics. Other skin conditions that can be improved by cos embodiments, the lotion contains water, mineral oil, isopro metic approaches include Scarring, e.g., from acne or other pyl myristate, PEG40 sorbitan peroleate, glyceryl lanolate, causes, uneven pigmentation, and the like. A variety of Sorbitol, propylene glycol, cetyl palmitate, magnesium Sul procedures have been developed for improving skin appear fate, aluminum stearate, lanolin alcohol, BHT methylchlor ance. Examples of Such procedures include treatment with oisothiazolinone, and methylisothiazolinone. In some Botulinum Toxin Type A (“botox'), retinoids and derivatives embodiments, the lotion contains petrolatum or mineral oil, thereof and especially retinoic acid (all-trans and 13-cis) and a quaternary ammonium, a fatty alcohol, and a fattey ester retinol, and the use of hydroxy acids. Although progress has emollient. In some embodiments, the lotion contains water, been made in the use of cosmetic compositions for the glycerin, distearyldimonium chloride, petrolatum, isopropyl treatment of skin, response to treatment is variable and often palmitate, cetyl alcohol, dimethicone, Sodium chloride, a condition is only marginally to moderately responsive to methylparaben, and propylparaben. In some embodiments treatment. Unfortunately, once applied to the skin, Some of where the vehicle is a lotion, the lotion contains the TGF-B these agents can cause itching, stinging and tightness which mimic at a concentration of about 0.0001% to about 0.01% may lead to considerable discomfort. For many Subsects, by weight. In some embodiments where the vehicle is a sensitivity to Sun is enhanced. The use of these products by lotion, the lotion contains the TGF-B mimic at a concentra consumers with sensitive skin is often prevented. Many of tion of about 0.001% by weight. the same considerations apply to other methods for cosmetic treatment of skin, such as laser resurfacing or dermabrasion. 0006. In some embodiments of the methods of the inven Thus, there remains a need for cosmetic and/or dermato tion, the TGF-B mimic is administered in combination with logical compositions and methods for the treatment of skin. one or more other cosmetic or dermatological agents. In Some embodiments, the other cosmetic or dermatological SUMMARY OF THE INVENTION agent or agents is hydroxy acids, retinoic acid derivatives, 0002 The invention provides methods, compositions, free radical scavengers, botulinum toxin, Sunscreens, anti and kits for the cosmetic use of TGF-B mimics. acne agents, and anticellulite agents. 0003. In one aspect, the invention provides methods 0007. In another aspect, the invention provides compo utilizing TGF-B mimics. In some embodiments of this sitions. In some embodiments of this aspect, the composi aspect, the invention provides a method for cosmetic treat tions include a TGF-B mimic in a cosmetically acceptable ment of skin in an individual that includes topically admin vehicle at a concentration of greater than about 0.00005% by istering to an individual desiring or in need of cosmetic weight. In some embodiments of this aspect, the composi treatment an effective amount of a TGF-B mimic. In some tions include a TGF-B mimic at a concentration of about embodiments, the cosmetic treatment is used to modulate a 0.0001% to about 0.01% by weight. In some embodiments cosmetic condition Such as skin aging, cosmetic defect, of the invention, the vehicle includes a mixture of emulsi undesired pigmentation, or post-cosmetic procedure dam fying lanolin alcohols, waxes, and oils. In some of the age. In some embodiments, the cosmetic treatment is used to compositions of the invention, the vehicle includes water, augment an area of skin or epithelium (e.g., lips). In some mineral oil, isopropyl myristate, PEG-40 sorbitan peroleate, embodiments where the cosmetic condition is a cosmetic glyceryllanolate, Sorbitol, propylene glycol, cetyl palmitate, defect, the cosmetic defect can be, e.g., striae gravidorum, magnesium sulfate, aluminum Stearate, lanolin alcohol, striae distensiae, atrophic scarring, wound or Surgical scar BHT methylchloroisothiazolinone, methylisothiazolinone. ring, or hair loss. In some embodiments where the cosmetic In some embodiments, these compositions may include the condition is post-cosmetic procedure damage, the post TGF-B mimic at a concentration of about 0.001% by weight. cosmetic procedure damage can result from, e.g., chemical In some embodiments, the vehicle contains petrolatum or peel, dermabrasion, laser resurfacing, ablative resurfacing, mineral oil; a quaternary ammonium, a fatty alcohol, and a nonablative resurfacing, photodynamic therapy, noncoher fattey ester emollient. In some of the compositions of the ent light phototherapy, breast lift, face lift, eyelid lift, invention, the vehicle includes water, glycerin, distearyldi forehead lift, neck lift, thigh lift, buttock lift, tummy tuck, monium chloride, petrolatum, isopropyl palmitate, cetyl and scar revision. In some embodiments, the cosmetic alcohol, dimethicone, sodium chloride, methylparaben, and condition is skin aging. In some embodiments, the skin propylparaben. In some embodiments, these compositions aging can result in, e.g., Wrinkling, loss of elasticity, Sag may include the TGF-B mimic at a concentration of about ging, uneven pigmentation, and loss of underlying tissue 0.001% by weight. a SS. 0008. In yet another aspect, the invention provides kits 0004. In embodiments of methods of the invention, the for use in the cosmetic treatment of skin. In some embodi TGF-B mimic is administered at an average frequency of ments of this aspect of the invention, the kit includes a US 2006/0293227 A1 Dec. 28, 2006

composition containing a TGF-B mimic in a cosmetically conformation that results in an effect similar to a natural acceptable vehicle and instructions for use of the composi TGF-B, even if the conformation is not similar to that of a tion in the cosmetic treatment of skin. natural TGF-?3. INCORPORATION BY REFERENCE 0016 1. Structure and Activity 0017 Examples of TGF-B mimics are described in, e.g., 0009 All publications and patent applications mentioned U.S. Pat. Nos. 5,661,127: 5,780,436; and 6,638,912. These in this specification are herein incorporated by reference to include compounds that are structurally or biologically the same extent as if each individual publication or patent analogous to a region of TGF-B and mimic the conformation application was specifically and individually indicated to be recognized by TGF-B binding species (e.g., TGF-B recep incorporated by reference. tors). BRIEF DESCRIPTION OF THE DRAWINGS 0018 FIGS. 1A and 1B set out the coordinates for the atoms in a structure defining the biologically active TGF-B 0010. The novel features of the invention are set forth mimics useful in the methods of the invention. The coordi with particularity in the appended claims. A better under nate computations were carried out using as a model com standing of the features and advantages of the present pound, cytomodulin, described in U.S. Pat. No. 5,661,127. invention will be obtained by reference to the following having the amino acid sequence Ala-Asn-Val-Ala-Glu-Asn detailed description that sets forth illustrative embodiments, Ala (SEQID NO:1); these coordinates were obtained by use in which the principles of the invention are utilized, and the of the AMBER and MIDAS programs. Structures with accompanying drawings of which: substantially these same coordinates (by “substantially” is 0011 FIGS. 1A and 1B depict the spatial coordinates for meant to include up to about a 15% variance) will generate the atoms in a structure defining the biologically active the desired surfaces and generally display the biological activities useful in the methods of the invention; presence of TGF-B mimics useful in the methods of the invention. biological activity can be confirmed by the assays discussed below. The allowance for an amount of some variance is due DETAILED DESCRIPTION OF THE to mobility and adaptability of fit between ligand and TGF-B INVENTION receptor. Thus, such structures are considered to be TGF-B 0012. The present invention provides methods, compo mimics for purposes of this invention. sitions, and kits for the cosmetic use of transforming growth factor-beta (TGF-B) mimics. In one aspect, the invention 0019 Assays that may be used to assess TGF-3 activity provides methods of cosmetic application of TGF-B mimics include those that measure the ability to promote anchorage for cosmetic treatment of skin in an individual by topically independent growth of normal fibroblasts, for example, the administering an effective amount of a TGF-B mimic to the growth and colony formation by NRK-49 F fibroblasts in individual. In embodiments of this aspect, the cosmetic soft agar. Other assays that may be used to determine TGF-B treatment is used to modulate a cosmetic condition Such as activity include the inhibition of DNA synthesis in MV-1-Lu skin aging (which can include wrinkling, uneven pigmen mink lung epithelial cells, the induction of increased expres tation, sagging, and the like), cosmetic defect, undesired sion of type I collagen in primary cultures of neonatal human pigmentation, post-cosmetic procedure damage, and scar dermal fibroblasts, and/or induction of TGF-B synthesis. ring due to a skin condition. In some embodiments, the 0020 Several amino acid sequences have been found to TGF-B mimic is used in combination with other cosmetic or be TGF-B mimics, and are exemplary of TGF-B mimics dermatological agents or methods. useful in the methods provided herein. ATGF-B mimic thus 0013 In another aspect, the invention provides compo may include an amino acid sequence as defined below. sitions suitable for topical cosmetic use that include a TGF-B 0021. The term "amino acid as used herein means an mimic. In some embodiments the compositions include one organic compound containing both a basic amino group and or more additional components besides a TGF-B mimic. In an acidic carboxyl group. Included within this term are still another aspect, the invention provides kits for the natural amino acids (e.g., generally the L-amino acids), and cosmetic application of TGF-B) mimics to skin. amino acids and imino acids which are known to occur biologically in free or combined form but usually do not 0014 I. TGF-B Mimics occur in proteins. Included within this term also are modified 0.015 The methods and compositions of the invention and unusual amino acids, such as those disclosed in, for relate to TGF-B mimics. A “TGF-B mimic, as that term is example, Roberts and Vellaccio (1983) The Peptides, 5: used herein, includes synthetic or naturally-occurring com 342-429 (e.g., D-amino acids). In addition, the term "amino pounds. It can be any Suitable molecule. Such as a peptide, acid also includes other non-naturally occurring amino peptidomimetic, peptide nucleic acid, antibody, or Small or acids besides the D-amino acids, which are functional large inorganic or organic molecule, or a mixture of two or equivalents of the naturally-occurring amino acids. Such more of these types of compounds (e.g., compound that non-naturally-occurring (also referred to herein as "unnatu contains peptide and peptidomimetic portions), that is ral amino acids) amino acids include, for example, norleu capable of assuming a conformation that causes it to have at cine (“Nle'), norvaline (“Nva'), B-Alanine, L- or D-naph least one activity of TGF-B, as measured by one or more of thalanine, ornithine ("Orn'), homoarginine (homoArg) and the assays described herein. This conformation may be a others well known in the peptide art, such as those described conformation that is Substantially similar to or the same as in M. Bodanzsky, Principles of Peptide Synthesis, 1st and the structure of a region of TGF-B, as discussed more fully 2nd revised ed., Springer-Verlag, New York, N.Y., 1984 and below. Alternatively, this conformation may be any suitable 1993, and Stewart and Young, Solid Phase Peptide Synthe US 2006/0293227 A1 Dec. 28, 2006

sis, 2nd ed., Pierce Chemical Co., Rockford, Ill., 1984. 2-amino-2-(cyclohexenyl)acetic acid, 2-amino-3-(2-cyclo Amino acids and amino acid analogs can be purchased pentenyl)propanoic acid, 2-amino-3-(3-cyclopentenyl)pro commercially (Sigma Chemical Co.; Advanced Chemtech; panoic acid, 2-amino-3-(1-cyclohexyl)propanoic acid, RSP; Bachem; or ChemImpex) or synthesized using meth 2-amino-2-(1-cyclopentenyl)acetic acid, 2-amino-2-(1-cyl ods known in the art. cohexyl)acetic acid, 2-amino-2-(1-cylcoheptenyl)acetic 0022 "Natural amino acids include, but are not limited acid, 2-amino-2-(1-cyclooctenyl)acetic acid, 2-amino-3-(1- to, alanine, arginine, asparagine, aspartic acid, cysteine, cycloheptenyl)propanoic acid, 2-amino-3-(1,4-cyclohexadi glutamic acid, glutamine, glycine, histidine, isoleucine, leu enyl)propanoic acid, 2-amino-3-(2,5-cyclohexadienyl)pro cine, lysine, methionine, phenylalanine, serine, threonine, panoic acid, 2-amino-2-(7-cycloheptatrienyl)acetic acid, tyrosine, tyrosine, tryptophan, proline, and valine. Natural 2-amino-4,5-hexadienoic acid, 2-amino-3-butynoic acid, non-protein amino acids include, but are not limited to 2-amino-4-pentyoic acid, 2-amino-4-hexynoic acid, arginosuccinic acid, citruline, cysteine Sulfinic acid, 3,4- 2-amino-4-hepten-6-ynoic acid, 2-amino-3-fluoropropanoic dihydroxyphenylalanine, homocysteine, homoserine, orni acid, 2-amino-3,3,3-trifluoropropanoic acid, 2-amino-3- thine, 3-monoiodotyrosine, 3,5-diiodotryosine, 3.5.5'-tri fluorobutanoic acid, 2-amino-3-fluoropentanoic acid, iodothyronine, and 3.3',5,5'-tetraiodothyronine. Modified or 2-amino-3-fluorohexanoic acid, 2-amino-3,3-difluorobu unusual amino acids which can be used to practice the tanoic acid, 2-amino-3,3-difluoro-3-phenylpropanoic acid, invention include, but are not limited to, D-amino acids, 2-amino-3-perfluoroethylpropanoic acid, 2-amino-3-per hydroxylysine, 4-hydroxyproline, an N-CBZ-protected fluoropropylpropanoic acid, 2-amino-3-fluoro-3-methylbu amino acid, 2,4-diaminobutyric acid, homoarginine, norleu tanoic acid, 2-amino-5.5.5-trifluoropentanoic acid, 2-amino cine, N-methylaminobutyric acid, naphthylalanine, phenylg 3-methyl-4,4,4-trifluorobutanoic acid, 2-amino-3- lycine, B-phenylproline, tert-leucine, 4-aminocyclohexyla trifluoromethyl-4,4,4-trifluorobutanoic acid, 2-amino-3,3,4, lanine, N-methyl-norleucine, 3,4-dehydroproline, N.N- 4.5.5-heptafluoropentanoic acid, 2-amino-3-methyl-5- dimethylaminoglycine, N-methylaminoglycine, fluoropentanoic acid, 2-amino-3-methyl-4-fluoropentanoic 4-aminopiperidine-4-carboxylic acid, 6-aminocaproic acid, acid, 2-amino-5,5-difluorohexanoic acid, 2-amino-4-(fluo trans-4-(aminomethyl)-cyclohexanecarboxylic acid, 2-, 3-, romethyl)-5-fluoropentanoic acid, 2-amino-4-trifluorom and 4-(aminomethyl)-benzoic acid, 1-aminocyclopentan ethyl-5.5.5-trifluoropentanoic acid, 2-amino-3-fluoro-3-me ecarboxylic acid, 1-aminocyclopropanecarboxylic acid, and thylbutanoic acid, 2-amino-3-fluoro-3-phenylpentanoic 2-benzyl-5-aminopentanoic acid. acid, 2-amino-2-(1-fluorocyclopentyl)acetic acid, 2-amino 2-(1-fluorocyclohexyl)acetic acid, 2-amino-3-chloropro 0023 Standard three- and one-letter abbreviations for panoic acid acid, 2-amino-3-chlorobutanoic acid acid, natural amino acid residues or amino acids apply throughout 2-amino-4,4-dichlorobutanoic acid acid, 2-amino4.4.4- the specification unless otherwise indicated. trichlorobutanoic acid, 2-amino-3,4,4-trichlorobutanoic 0024. Unnatural amino acids that fall within the scope of acid, 2-amino-6-chlorohexanoic acid, 2-amino-4-bromobu this invention are by way of example and without limitation: tanoic acid, 2-amino-3-bromobutanoic acid, 2-amino-3-mer 2-aminobutanoicacid, 2-aminopentanoic acid, 2-aminohex captobutanoic acid, 2-amino-4-mercaptobutanoic acid, anoic acid, 2-aminoheptanoicacid, 2-aminooctanoic acid, 2-amino-3-mercapto-3,3-dimethylpropanoic acid, 2-amino 2-aminononanoic acid, 2-aminodecanoic acid, 2-aminoun 3-mercapto-3-methylpentanoic acid, 2-amino-3-mercapto decanoic acid, 2-amino-3,3-dimethylbutanoic acid, pentanoic acid, 2-amino-3-mercapto-4-methylpentanoic 2-amino-4,4-dimethylpentanoic acid, 2-amino-3-methyl acid, 2-amino-3-methyl-4-mercaptopentanoic acid, hexanoic acid, 2-amino-3-methylheptanoic acid, 2-amino 2-amino-5-mercapto-5-methylhexanoic acid, 2-amino-2-(1- 3-methyloctanoic acid, 2-amino-3-methylnonanoic acid, mercaptocyclobutyl)acetic acid, 2-amino-2-(1-mercaptocy 2-amino-4-methylhexanoic acid, 2-amino-3-ethylpentanoic clopentyl)acetic acid, 2-amino-2-(1-mercaptocyclohexy acid, 2-amino-3,4-dimethylpentanoic acid, 2-amino-3,5- l)acetic acid, 2-amino-5-(methylthio)pentanoic acid, dimethylhexanoic acid, 2-amino-3,3-dimethylpentanoic 2-amino-6-(methylthio)hexanoic acid, 2-amino-4-meth acid, 2-amino-3-ethyl-3-methylpentanoic acid, 2-amino-3, ylthio-3-phenylbutanoic acid, 2-amino-5-ethylthio-5-meth 3-diethylpentanoic acid, 2-amino-5-methylhexanoic acid, ylpentanoic acid, 2-amino-5-ethylthio-3.5.5-trimethylpen 2-amino-6-methylheptanoic, 2-amino-7-methyloctanoic, tanoic acid, 2-amino-5-ethylthio-5-phenylpentanoic acid, 2-amino-2-cyclopentylacetic, 2-amino-2-cylcohexylacetic 2-amino-5-ethylthio-5-pentanoic acid, 2-amino-5-butylthio acid, 2-amino-2-(1-methylcylcohexyl)acetic acid, 2-amino 5-methylpentanoic acid, 2-amino-5-butylthio-3.5.5-trimeth 2-(2-methyl-1-methylcylcohexyl)acetic acid, 2-amino-2-(3- ylpentanoic acid, 2-amino-5-butylthio-5-phenylpentanoic methyl-1-methylcylcohexyl)acetic acid, 2-amino-2-(4-me acid, 2-amino-5-(butylthio)pentanoic acid, 2-amino-3-me thyl-methylcylcohexyl)acetic acid, 2-amino-2-(1- thy4-hydroselenopentanoic acid, 2-amino-4-methylsele ethylcycolhexyl)acetic acid, 2-amino-3- nobutanoic acid, 2-amino-4-ethylselenobutanoic acid, (cyclohexyl)propanoic acid, 2-amino-4- 2-amino-4-benzylselenobutanoic acid, 2-amino-3-methyl-4- (cyclohexyl)butanoic acid, 2-amino-3-(1- (methylseleno)butanoic acid, 2-amino-3-(aminomethylsele adamantyl)propanoic acid, 2-amino-3-butenoic acid, no)propanoic acid, 2-amino-3-(3-aminopropylseleno)pro 2-amino-3-methyl-3-butenoic acid, 2-amino-4-pentenoic panoic acid, 2-amino-4-methyltellurobutanoic acid, acid, 2-amino-4-hexenoic acid, 2-amino-5-heptenoic acid, 2-amino-4-hydroxybutanoic acid, 2-amino-4-hydroxyhex 2-amino-4-methyl-4-hexenoic acid, 2-amino-5-methyl-4- anoic acid, 2-amino-3-hydroxypentanoic acid, 2-amino-3- hexenoic acid, 2-amino-4-methy-5-hexenoic acid, 2-amino hydroxyhexanoic acid, 2-amino-3 methyl-4-hydroxybu 6-heptenoic acid, 2-amino-3,3,4-trimethyl-4-pentenoic acid, tanoic acid, 2-amino-3-hydroxy-3-methylbutanoic acid, 2-amino-4-chloro-4-pentenoic, 2-amino-4,4-dichloro-3- 2-amino-6-hydroxyhexanoic acid, 2-amino-4-hydroxyhex butenoic acid, 2-amino-3-(2-methylenecyclopropyl)-pro anoic acid, 2-amino-3-hydroxy-4-methylpentanoic acid, panoic acid, 2-amino-2-(2-cyclopentenyl)acetic acid, 2-amino-3-hydroxy-3-methylpentanoic acid, 2-amino4-hy

US 2006/0293227 A1 Dec. 28, 2006

indolyl)propanoic acid, 2-amino-3-(7-nitro-3- l)acetic acid, 2-amino-2-(2-thienyl)acetic acid, 2-amino-2- indolyl)propanoic acid, 2-amino-3-(4-carboxy-3- (2-thiazolyl)acetic acid, 2-amino-3-(2-thiazolyl)propanoic indolyl)propanoic acid, 2-amino-3-(3-indolyl)butanoic acid, acid, 2-amino-4-(4-carboxy-2-thiazolyl)butanoic acid, 2-amino-3-(2,3-dihydro-3-indolyl)propanoic acid, 2-amino 2-amino-3-(4-thiazolyl)propanoic acid, 2-amino-3-(2-sele 3-(2,3-dihydro-2-oxo-3-indolyl)propanoic acid, 2-amino-3- nolyl)propanoic acid, 2-amino-3-(2-amino-4-selenolyl)pro alkylmercapto-3-(3-indolyl)propanoic acid, 2-amino-3-(4- panoic acid, and 2-amino-3-(beta-ribofuranosyl)propanoic aza-3-indolyl)propanoic acid, 2-amino-3-(7-aza-3- acid. indolyl)propanoic acid, 2-amino-3-(7-aza-6-chloro-4- 0025 “Amino acids residue' has its customary meaning methyl-3-indolyl)propanoic acid, 2-amino-3-(2,3- in the art and refers to an amino acid that is part of a peptide dihydrobenzofuran-3-yl)propanoic acid, 2-amino-3-(3- or polypeptide chain; “amino acid residue' as used herein methyl-5-7-dialkylbenzofuran-2-yl)propanoic acid, also refers to various amino acids where sidechain func 2-amino-3-(benzothiophen-3-yl)propanoic acid, 2-amino-3- tional groups are coupled with appropriate protecting groups (5-hydroxybenzothiophen-3-yl)propanoic acid, 2-amino-3- known to those skilled in the art. “The Peptides”, Vol 3, 3-88 eoenzoselenol-3-yl)propanoic acid, 2-amino-3-quinolylpro (1981) discloses numerous Suitable protecting groups. panoic acid, 2-amino-3-(8-hydroxy-5-quinolyl)propanoic Examples of amino acids where sidechain functional groups acid, 2-amino-2-(5,6,7,8-tetrahydroquinol-5-yl)acetic acid, are coupled with appropriate protecting groups include, but 2-amino-3-(3-coumarinyl)propanoic acid, 2-amino-2-(ben are not limited to, Asp(OMe), GlucOMe), Hyp(OMe), Zisoxazol-3-yl)acetic acid, 2-amino-2-(5-methylbenzisoX Asp(O'Bu), GlucC'Bu), Hyp(O'Bu), Thr(O'Bu), Asp(OBzl), aZol-3-yl)acetic acid, 2-amino-2-(6-methylbenzisoxazol-3- Glu(OBzl), Hyp(OBzl), and Thr(OBzl). yl)acetic acid; 2-amino-2-(7-methylbenzisoxazol-3- 0026. Many TGF-B mimics are composed of, or include, yl)acetic acid, 2-amino-2-(5-bromobenzisoxazol-3-yl)acetic a peptide. The term "peptide' as used herein refers to acid, 2-amino-3-(benzimidazol-2-yl)propanoic acid, polymers of amino acids under 100 amino acids in length; in 2-amino-3-(5,6-dichlorobenzimidazol-2-yl)propanoic acid, preferred embodiments, less than 30 amino acids in length. 2-amino-3-(5,6-dimethylbenzimidazol-2-yl)propanoic acid, A peptide may be more than about 1, 2, 3, 4, 5, 6, 7, 8, 9. 2-amino-3-(4,5,6,7-hydrobenzirnidazol-2-yl)propanoic or 10 amino acids in length. A peptide may be less than 100, acid, 2-amino-2-(benzimidazol-5-yl)acetic acid, 2-amino-2- 90, 80, 70, 60, 50, 40, 35, 30, 25, 20, 15, 14, 13, 12, 11, 10, (1,3-dihydro-2,2-dioxoisobenzothiophen-5-yl)acetic acid, 9, 8, 7, 6, 5, 4, or 3 amino acids in length. Preferred peptides 2-amino-2-(1,3-dihydro-2,2-dioxo-2,1,3-benzothiadiazol-5- are less than 30 amino acids in length. The polymer may be yl)acetic acid, 2-amino-2-(2-oxobenzimidazol-5-yl)acetic linear or branched, it may comprise amino acids of any type acid, 2-amino-3-(4-hydroxybenzothiazol-6-yl)propanoic as defined above, and it may be interrupted by non-amino acid, 2-amino-3-(benzoxazol-2-yl)propanoic acid, 2-amino acids (e.g., peptidomimetics). The term also encompasses an 3-(benzothiazol-2-yl)propanoic acid, 2-amino-3-(9-adeni amino acid polymer that has been modified naturally or by nyl)propanoic acid, 2-amino-2-(6-chloro-9-purinyl)acetic intervention; for example, disulfide bond formation, glyco acid, 2-amino-2-(6-amino-9-purinyl)acetic acid, 2-amino-3- Sylation, lipidation, acetylation, carboxylation, phosphory (6-purinyl)propanoic acid, 2-amino-3-(8-theobrominyl)pro lation, ubiquitination, pegylation or any suitable other panoic acid, 2-amino-2-(1-uracilyl)acetic acid, 2-amino-2- manipulation or modification, Such as conjugation with a (1-cytosinyl)acetic acid, 2-amino-3-(1-uracilyl)propanoic labeling component. acid, 2-amino-3-(1-cytosinyl)propanoic acid, 2-amino-4-(1- pyrimidinyl)butanoic acid, 2-amino-4-(4-amino-1-pyrimidi 0027. The hydrophobicity, charge, ability to form hydro nyl)butanoic acid, 2-amino-4-(4-hydroxy-1-pyrimidinyl)bu gen bonds, and other properties of amino acids can be tanoic acid, 2-amino-5-(1-pyrimidinyl)pentanoic acid, involved in producing peptides that act as TGF-B mimics. Of 2-amino-5-(4-amino-1-pyrimidinyl)pentanoic acid, the twenty common amino acids, those with hydrophobic R 2-amino-5-(4-hydroxy-1-pyrimidinyl)pentanoic acid, groups include: Alanine, Valine, Leucine, Isoleucine, Pro 2-amino-3-(5-pyrimidinyl)propanoic acid, 2-amino-3-(6- line, Phenylalanine, Tryptophan and Methionine. Amino uracilyl)propanoic acid, 2-amino-3-(2-pyrimidinyl)pro acids with uncharged polar R groups include: Glycine, panoic acid, 2-amino-3-(6-amino-4-chloro-2-pyrimidinyl Serine, Threonine, Cysteine, Tyrosine, Asparagine and )propanoic acid, 2-amino-3-(4-hydroxy-2- Glutamine. Amino acids with charged polar R groups pyrimidinyl)propanoic acid, 2-amino-3-(2-amino-4- include: Aspartic acid and Glutamic acid. Basic amino acids pyrimidinyl)propanoic acid, 2-amino-3-(4.5- include: Lysine, Arginine and Histidine (at pH 6.0). Amino dihydroxypyrimidin-2-yl)propanoic acid, 2-amino-3-(2- acids with phenyl groups include: Phenylalanine, Tryp thiouracil-6-yl)propanoic acid, 2-amino-2-(5-alkyl-2- tophan and Tyrosine. As will be apparent to those of skill in tetrahydrofuryl)acetic acid, 2-amino-2-(5-methyl-2,5- the art, many unnatural or artificial amino acids also fall dihydro-2-furyl)acetic acid, 2-amino-2-(5-alkyl-2- within the various types of amino acids, and may be used in furyl)acetic acid, 2-amino-2-(2-furyl)acetic acid, 2-amino embodiments of the invention. 2-(3-hydroxy-5-methyl-4-isoxazolyl)acetic acid, 2-amino 3-(4-bromo-3-hydroxy-5-isoxazolyl)propanoic acid, 0028. In exemplary peptides useful in the invention, the 2-amino-3-(4-methyl-3-hydroxy-5-isoxazolyl)propanoic appropriate functional groups are represented, in many acid, 2-amino-3-(3-hydroxy-5-isoxazolyl)propanoic acid, cases, within the following amino acid sequence, which may 2-amino-2-(3-chloro-D2-isoxazolin-5-yl)acetic acid, be the entire TGF-B mimic or may be included in a larger 2-amino-2-(3-oxo-5-isoxazolidinyl)acetic acid, 2-amino-3- structure: AA-AA-AA. . . . -AA. In some embodi (3,5-dioxo-1,2,4-oxadiazolin-2-yl)propanoic acid, 2-amino ments, TGF-B mimics useful in the methods of the invention 3-(3-phenyl-5-isoxazolyl)propanoic acid, 2-amino-3-3-(4- contain: hydroxyphenyl)-1,2,4-oxadiazol-5-ylpropanoic acid, 0029 (1) a hydrophobic or neutral amino acid at 2-amino-3-(2-thienyl)propanoic acid, 2-amino-2-(2-fury position i; US 2006/0293227 A1 Dec. 28, 2006

0030 (2) a branched hydrophobic at position i-1 (e.g., 0037 As discussed above, the original peptide discov Val, Ile): ered to have TGF-B activity has been named “cytomodulin' 0031 (3) a small aliphatic at position i+2 (e.g., Ala), and has the sequence A-N-V-A-E-N-A (SEQ ID NO: 1). where positions i+1 and i+2 together form a structure Cytomodulin when added to cells in culture in the concen found in TGF-B mimics, the U-bend (B-bend) structure: tration range 10 to 10 M (1.4 pg/mL to 1400 pg/mL). and elicits certain highly specific TGF-B effects in several dif ferent cell types. For example, among the effects observed is 0032 (4) a side-chain containing a hydrogen-bond the inhibition of DNA synthesis in MV-1-Lu mink lung acceptor shortly thereafter (at position AA). epithelial cells, the growth and colony formation by NRK 0033 For example, in some embodiments, AA, is alanine, 49 F fibroblasts in soft agar, and the induction of increased asparagine, or leucine, AA is valine or isoleucine, and expression of type I collagen in primary cultures of neo AA is alanine. One aspect of TGF-B mimics useful in the natal human dermal fibroblasts. Moreover, results with invention is the relative positioning of the side-chains of human osteogenic sarcoma (HOS) cell line indicate that AA, and AA. The correct positioning of these amino cytomodulin also may be a mimic for other members of the acids can be achieved if AA and AA are in either of two TGF-B Superfamily. Such as bone morphogenic proteins backbone conformations: a B-bend or an “extended-bend (BMPs) and osteogenic protein (OPs), as evidenced by its conformation with the backbone (d.1) angles of AA ability to specifically stimulate markers (alkaline phos equal to approximately (-60, +135) and those of AA equal phatase and osteonectin) characteristic of the osteoblast to approximately (-60, -45). This sequence is often either phenotype. immediately followed by or has proximal thereto an amino acid with a hydrogen bond acceptor-containing side-chain. 0038) Other exemplary TGF-B mimics useful in the Because the amino acid residue with a hydrogen bond methods of the invention are described in U.S. Pat. No. acceptor-containing side-chain does not necessarily have to 6,638.912 and are also described in the Examples. be immediately followed by, that is adjacent to AA, it is 0039) Peptide TGF-B mimics useful in the invention referred to as AA where n is an integer equal to or greater include peptides having at least 70%, at least 80%, at least than three. If n is greater than 3, then n-3 (“n minus 3') 90%, at least 95% or greater than 95% sequence identity to amino acid residues would be between AA and AA in the amino acid sequences disclosed herein. In some embodi the peptide sequence. ments, the invention provides methods and compositions 0034) For example, the TGF-B mimics may include the that utilize a peptide having at least 70%, at least 80%, at sequence AA-AA-AA-AA, where AA, through AA, least 90%, at least 95% or greater than 95% sequence are as before described and AA is an amino acid residue identity to an amino acid sequence comprising one of the with a hydrogen bond acceptor-containing side-chain (and is amino acid sequences of SEQ ID NOS: 1-42. In some glutamic acid, aspartic acid, glutamine, or asparagine). The embodiments, the invention provides methods and compo original peptide with TGF-B activity and an especially sitions that utilize a peptide having at least 70%, at least preferred TGF-B mimic for use in the methods of the present 80%, at least 90%, at least 95% or greater than 95% invention, cytomodulin, A-N-V-A-E-N-A (SEQID NO: 1) is sequence identity to an amino acid sequence comprising one of this class. Here, AA-AA-AA-AA corresponds to of the amino acid sequences of SEQID NOS: 1, 2, 3, 11, 13, the second through fifth residues from the N-terminus of or 14. In some embodiments, the invention provides meth cytomodulin, -N-V-A-E-. Other preferred embodiments are ods and compositions that utilize a peptide having at least the peptides, L-I-A-E-A-K (SEQ ID NO:2) and L-I-A-E- 70%, at least 80%, at least 90%, at least 95% or greater than A-A (SEQID NO: 11). In these examples, AA-AA-AA 95% sequence identity to the amino acid sequence of SEQ 2-AA corresponds to the first four residues of the peptide, ID NO: 1. L-I-A-E-. 0040 Percent sequence identity is determined by con 0035) Another example is the sequence AA-AA-AA ventional methods. See, for example, Altschul et al., Bull. 2-AA-AA where the residue with the hydrogen bond Math. Bio. 48:603 (1986), and Henikoff and Henikoff, Proc. acceptor-containing side-chain is not immediately adjacent, Natl. Acad. Sci. USA 89:10915 (1992). Briefly, two amino but instead is proximal to, the initial sequence. In this case, acid sequences are aligned to optimize the alignment scores AA, through AA are as before, AA is any suitable using a gap opening penalty of 10, a gap extension penalty amino acid, and AA may be glutamic acid, aspartic acid, of 1, and the “BLOSUM62 scoring matrix of Henikoff and glutamine, or asparagine. A preferred embodiment of this Henikoff (ibid.). The percent identity is then calculated as: class is L-I-A-G-E-G (SEQ ID NO: 14). An especially (Total number of identical matches/length of the longer preferred embodiment is the peptide, L-I-A-P-E-A (SEQ ID sequence plus the number of gaps introduced into the longer NO:3). In both examples, the first five N-terminal amino sequence in order to align the two sequences)(100). acids correspond to AA-AA-AA-AAis-AA. 0041 Those skilled in the art appreciate that there are 0.036 Yet another example is the sequence AA-AA many established algorithms available to align two amino AA-AAs-AA-AAs. Here, AA, through AA-2 are as acid sequences. The “FASTA similarity search algorithm of before, AA and AA are suitable amino acids, and AAs Pearson and Lipman is a Suitable protein alignment method may be glutamic acid, aspartic acid, glutamine, or aspar for examining the level of identity shared by an amino acid agine. An especially preferred member of this series is the sequence disclosed herein and the amino acid sequence of a peptide, L-I-A-G-G-E (SEQ ID NO: 13). In this particular putative MMP-1 variant. The FASTA algorithm is described example there is a one to one correspondence between SEQ by Pearson and Lipman, Proc. Nat'l Acad. Sci. USA 85:2444 ID NO:3 and the sequence AA-AA-AA-AA-AA (1988), and by Pearson, Meth. Enzymol. 183:63 (1990). 4-AAs. Briefly, FASTA first characterizes sequence similarity by US 2006/0293227 A1 Dec. 28, 2006

identifying regions shared by the query sequence (e.g., SEQ “conservative amino acid substitution' preferably refers to a ID NO: 1) and a test sequence that have either the highest substitution represented by a BLOSUM62 value of greater density of identities (if the ktup variable is 1) or pairs of than -1. For example, an amino acid Substitution is conser identities (if ktup=2), without considering conservative vative if the substitution is characterized by a BLOSUM62 amino acid substitutions, insertions, or deletions. The ten value of 0, 1, 2, or 3. According to this system, preferred regions with the highest density of identities are then res conservative amino acid Substitutions are characterized by a cored by comparing the similarity of all paired amino acids BLOSUM62 value of at least 1 (e.g., 1, 2 or 3), while more using an amino acid Substitution matrix, and the ends of the preferred conservative amino acid Substitutions are charac regions are “trimmed to include only those residues that terized by a BLOSUM62 value of at least 2 (e.g., 2 or 3). contribute to the highest score. If there are several regions with scores greater than the “cutoff value (calculated by a 0043. It also will be understood that amino acid predetermined formula based upon the length of the sequences may include additional residues, such as addi sequence and the ktup value), then the trimmed initial tional N- or C-terminal amino acids, and yet still be essen regions are examined to determine whether the regions can tially as set forth in one of the sequences disclosed herein, be joined to form an approximate alignment with gaps. So long as the sequence retains Sufficient biological protein Finally, the highest scoring regions of the two amino acid activity to be functional in the compositions and methods of sequences are aligned using a modification of the Needle the invention. man-Wunsch-Sellers algorithm (Needleman and Wunsch, J. 0044) A specialized kind of insertional variant is the Mol. Biol. 48:444 (1970); Sellers, SIAM J. Appl. Math. fusion protein. It is contemplated that the entire TGF-B 26:787 (1974)), which allows for amino acid insertions and mimic peptide or a fragment of the TGF-B mimic peptide deletions. Illustrative parameters for FASTA analysis are: may be used to construct a fusion protein to enhance tissue ktup=1, gap opening penalty=10, gap extension penalty=1. specific or cell specific functions of the TGF-3 mimic and substitution matrix=BLOSUM62. These parameters can peptide useful in the invention. be introduced into a FASTA program by modifying the 0045. A fusion protein generally has all or a substantial scoring matrix file (“SMATRIX'), as explained in Appendix portion of the native molecule, linked at the N- or C-termi 2 of Pearson, Meth. Enzymol. 183:63 (1990). nus, to all or a portion of a second polypeptide. For example, 0042. The present invention also includes peptides hav fusions typically employ leader sequences from other spe ing a conservative amino acid change, compared with an cies to permit the recombinant expression of a protein in a amino acid sequence disclosed herein. Many such changes heterologous host. Another useful fusion includes the addi have been described specifically. More generally, for tion of an immunologically active domain, such as an example, variants can be obtained that contain one or more antibody epitope, to facilitate purification of the fusion amino acid substitutions of SEQ ID NO:1-42, in which an protein. Inclusion of a cleavage site at or near the fusion alkyl amino acid is substituted for an alkyl amino acid in a junction will facilitate removal of the extraneous polypep TGF-B mimic peptide amino acid sequence, an aromatic tide after purification. Other useful fusions include linking amino acid is substituted for an aromatic amino acid in a of functional domains, such as active sites from enzymes TGF-B mimic peptide amino acid sequence, a Sulfur-con Such as a hydrolase, glycosylation domains, cellular target taining amino acid is substituted for a Sulfur-containing ing signals or transmembrane regions. amino acid in a TGF-B mimic peptide amino acid sequence, a hydroxy-containing amino acid is Substituted for a 0046 2. Synthesis hydroxy-containing amino acid in a TGF-B mimic peptide 0047 For TGF-3 mimics which consist of or include amino acid sequence, an acidic amino acid is substituted for peptides, the peptide may be synthesized by any Suitable an acidic amino acid in a TGF-B mimic peptide amino acid method for producing peptides of a given sequence. Pref sequence, a basic amino acid is Substituted for a basic amino erably, peptides of the present invention can be synthesized acid in TGF-B mimic peptide amino acid sequence, or a by various suitable methods that are well known in the art, dibasic monocarboxylic amino acid is substituted for a preferably by Solid phase synthesis, manual or automated, as dibasic monocarboxylic amino acid in a TGF-3 mimic first developed by Merrifield and described by Stewart et al. peptide amino acid sequence. Among the common amino in Solid Phase Peptide Synthesis (1984). Chemical synthesis acids, for example, a "conservative amino acid Substitution” joins the amino acids in the predetermined sequence starting is illustrated by a substitution among amino acids within at the C-terminus. Basic Solid phase methods require cou each of the following groups: (1) glycine, alanine, Valine, pling the C-terminal protected C-amino acid to a Suitable leucine, and isoleucine, (2) phenylalanine, tyrosine, and insoluble resin Support. Amino acids for synthesis require tryptophan, (3) serine and threonine, (4) aspartate and protection on the C-amino group to ensure proper peptide glutamate, (5) glutamine and asparagine, and (6) lysine, bond formation with the preceding residue (or resin Sup arginine and histidine. The BLOSUM62 table is an amino port). Following completion of the condensation reaction at acid substitution matrix derived from about 2,000 local the carboxyl end, the C-amino protecting group is removed multiple alignments of protein sequence segments, repre to allow the addition of the next residue. Several classes of senting highly conserved regions of more than 500 groups of C-protecting groups have been described, see Stewart et al. related proteins (Henikoff and Henikoff, Proc. Nat'l Acad. in Solid Phase Peptide Synthesis (1984), with the acid labile, Sci. USA 89:10915 (1992)). Accordingly, the BLOSUM62 urethane-based tertiary-butyloxycarbonyl (Boc) being the Substitution frequencies can be used to define conservative historically preferred. Other protecting groups, and the amino acid substitutions that may be introduced into the related chemical strategies, may be used, including the base amino acid sequences of the present invention. Although it labile 9-fluorenylmethyloxycarbonyl (FMOC). Also, the is possible to design amino acid substitutions based solely reactive amino acid sidechain functional groups require upon chemical properties (as discussed above), the language blocking until the synthesis is completed. The complex array US 2006/0293227 A1 Dec. 28, 2006

of functional blocking groups, along with strategies and not desire treatment but may nonetheless be in need of limitations to their use, have been reviewed by Bodansky in treatment). The term “treating or “treatment as used herein Peptide Synthesis (1976) and, Stewart et al. in Solid Phase includes achieving a cosmetic benefit. By cosmetic benefit is Peptide Synthesis (1984). meant any desired modulation of the cosmetic condition being treated. For example, in an individual with wrinkling, 0.048 Solid phase synthesis is initiated by the coupling of cosmetic benefit includes eradication or lessening of the the described C-terminal C-protected amino acid residue. appearance of wrinkling. Also, a cosmetic benefit is Coupling requires activating agents, such as dicyclohexy achieved with the eradication oramelioration of one or more carbodiimide (DCC) with or without 1-hydroxybenzo-tria of the psychological symptoms associated with the under Zole (HOBT), diisopropylcarbodiimide (DIIPC), or eth lying condition Such that an improvement is observed in the yldimethylaminopropylcarbodiimide (EDC). After coupling patient, notwithstanding the fact that the patient may still be the C-terminal residue, the C-amino protected group is affected by the cosmetic condition. For example, a TGF-B removed by trifluoroacetic acid (25% or greater) in dichlo mimic provides cosmetic benefit not only when a cosmetic romethane in the case of acid labile tertiary-butyloxycarbo defect is eradicated, but also when an improvement is nyl (Boc) groups. A neutralizing step with triethylamine observed in the individual with respect to the cosmetic (10%) in dichloro-methane recovers the free amine (versus defect and its attendant consequences, such as psychological the salt). After the C-terminal residue is added to the resin, consequences. In some cases, methods and compositions of the cycle of deprotection, neutralization and coupling, with the invention may be directed at achieving a prophylactic intermediate wash steps, is repeated in order to extend the benefit. A “prophylactic.' or “preventive' effect includes protected peptide chain. Each protected amino acid is intro prevention of a condition, retarding the progress of a con duced in excess (three to five fold) with equimolar amounts of coupling reagent in Suitable solvent. Finally, after the dition (e.g., skin aging), or decreasing the likelihood of completely blocked peptide is assembled on the resin Sup occurrence of a condition. As used herein, “treating” or port, reagents are applied to cleave the peptide form the resin “treatment includes prophylaxis. and to remove the side chain blocking groups. Anhydrous 0054 As used herein, the term “effective amount” hydrogen fluoride (HF) cleaves the acid labile tertiary encompasses an amount Sufficient to effect beneficial or butyloxycarbonyl (Boc) chemistry groups. Several nucleo desired cosmetic results. An effective amount can be admin philic scavengers, such as dimethylsulfide and anisole, are istered in one or more administrations. In terms of cosmetic included to avoid side reactions especially on side chain treatment, an “effective amount of a TGF-B mimic of the functional groups. invention is an amount that is sufficient to palliate, amelio rate, stabilize, reverse or slow the progression of a cosmetic 0049 Slight amino acid modifications to a peptide TGF-B condition, or to provide a desired effect Such as cosmetic mimic sequence will not affect the peptide's ability to form augmentation of a soft tissue. An "effective amount may be suitable TGF-B mimics. These modifications include tech of a TGF-3 mimic used alone or in conjunction with one or niques to confer resistance to enzymatic degradation Such as more agents used to modulate a cosmetic condition. adding blocking groups to both the N- and C-terminal residues. Another method for preventing degradation and 0055) 1. Methods of Cosmetic Treatment premature clearance by the renal system is the use of 0056. The skin is subject to a number of cosmetic con unnatural amino acid substitutes in the peptide sequence. ditions that result in alterations of function and/or appear For example, N-methyl-alanine is often substituted for ala ance that are considered undesirable, and whose manifesta nine and C.-amino isobutryic acid and C-aminobutyric acid tion can lead to psychological discomfort as well as, in some are substitutes for bulky hydrophobic amino acids. cases, physiological discomfort or harm. In some cases, 0050 Recombinant techniques, as known in the art, may although no defect is present, it is nonetheless desirable to also be used to produce peptides suitable for the methods of the individual to augment or alter the skin in Such a way as the invention. Naturally-occurring proteins may be cleaved to produce a cosmetically pleasing effect. to produce a desired TGF-B mimic. Methods of designing 0057 Exemplary cosmetic conditions that may be modu and screening Small molecules may also be used. Methods lated by the methods of the invention include, but are not to generate and screen peptidomimetics may also be useful limited to, skin aging, cosmetic defect, undesired pigmen in producing TGF-B mimics. Substances that are a mixture tation, and post-cosmetic procedure damage. of peptide and peptidormimetics may also be used. 0058 Skin aging includes chronological aging as well as 0051) II. Methods of Cosmetic Use photoaging, and may appear as wrinkling, lack of elasticity 0.052 The methods of the invention have wide applica (e.g. sagging), uneven pigmentation, thinning of the skin bility to cosmetic conditions. The mode of administration for and/or collagen so that veins and other underlying structures cosmetic applications is typically topical, but administration become more prominent, and the like. Skin aging is a major and dosage regimens will vary depending on the cosmetic example of a skin condition that involves a decrease in cell proliferation and in cell function. As used herein, “skin condition whose modulation is sought. aging refers to alterations in the appearance and function of 0053. The present invention provides methods, compo skin that occur with aging, such as wrinkling, loss of sitions, and kits for cosmetic use with individuals. The term elasticity, sagging, uneven pigmentation (e.g., “age spots or “individual' as used herein includes humans as well as other “liver spots”), and loss of underlying tissue mass. Such mammals. In some embodiments, the compositions, meth conditions may be accelerated and/or exacerbated by expo ods, and/or kits are used to provide a cosmetic treatment to Sure to ultraviolet radiation (photoaging) and other envi an invividual desiring and/or in need of cosmetic treatment ronmental conditions. With age and/or exposure to UV (e.g., young children Subject to burn or other scarring may radiation, the epidermis thins and the skin appendages US 2006/0293227 A1 Dec. 28, 2006 atrophy. Hair becomes sparse and sebaceous secretions 0064. In addition, the methods of the invention may be decrease, with consequent Susceptibility to dryness, chap employed to enhance and/or accelerate recovery from stan ping, and fissuring. The dermis diminishes with loss of dard cosmetic procedures, which are in themselves damag elastic and collagen fibers. Moreover, keratinocyte prolif ing to skin and/or underlying tissues, and which may take eration (which is indicative of skin thickness and skin undesirably lengthy periods of time for recovery and/or may proliferative capacity) decreases with age. produce Suboptimal results. Such procedures include chemi 0059 An increase in keratinocyte proliferation and col cal peel, dermabrasion, laser resurfacing, ablative resurfac lagen production is believed to counteract skin aging, i.e., ing, nonablative resurfacing, photodynamic therapy, nonco wrinkles, thickness, elasticity and repair. According to the herent light phototherapy, breast lift, face lift, eyelid lift, present invention, a TGF-B mimic can be used cosmetically forehead lift, neck lift, thigh lift, buttock lift, tummy tuck, and scar revision. As will be apparent to those of skill in the to counteract, at least for a time, the effects of aging on skin. art, Some of these procedures can require further skin A formulation containing a TGF-B mimic may be applied firming (e.g., "lifting procedures) while others are more topically in areas where it is desired to counteract skin aging. extensively damaging to the Surface of the skin and require 0060 Also included in the skin conditions that may be assistance for healing in a timely and optimal fashion (e.g., treated by the methods of the invention are cosmetic defects chemical peel, dermabrasion, ablative and non-ablative skin that, while not pathological or physiologically harmful, may resurfacing). In some embodiments, the methods of the nonetheless cause psychological distress, in some cases to invention provide a method for achieving firming and lifting the extreme. In these cases it is desirable to correct a of the eyelids; this may be done either in place of or in particular feature or features causing distress or, alterna conjunction with a conventional eyelid lift procedure. The tively, enhance a feature considered desirable. In addition to methods of the invention may be used in conjunction with skin aging, Such conditions include, e.g., striae gravidorum both types of procedures to enhance and/or accelerate heal and striae distensiae ('stretch marks), atrophic scarring (e.g., ing and recovery. acne scarring), wound (e.g., traumatic wounds, chronic wounds, or burn wounds) or Surgical scarring, thickened and 0065. In any of the methods of the invention, a single cracked skin (especially on the feet), and hair loss. TGF-B mimic may be used, or more than one TGF-B mimic 0061. In the latter embodiments, the invention relates to may be used. Any suitable TGF-B may be used in accor the use of TGF-B preparations to enhance hair growth. Cells dance with the description herein. In some embodiments, the from which the hair is produced grow in the bulb of the TGF-B mimic used is that of SEQ ID NO: 1. In some follicle. They are extruded in the form of fibers as the cells embodiments, the TGF-3 mimic used is that of SEQID NO: proliferate in the follicle. Hair “growth refers to the for 2. In some embodiments, the TGF-B mimic used is that of mation and elongation of the hair fiber by the dividing cells. SEQ ID NO: 3. In some embodiments, the TGF-B mimic In some embodiments, the methods of the invention provide used is that of SEQ ID NO: 11. In some embodiments, the a means for altering the dynamics of the hair growth cycle TGF-B mimic used is that of SEQ ID NO: 13. In some to induce proliferation of hair follicle cells, particularly stem embodiments, the TGF-B mimic used is that of SEQID NO: cells of the hair follicle. The subject compositions and 14. In some embodiments, a TGF-B of any of SEQID NOS: method can be used to increase hair follicle size and the rate 4-10, 12, or 15-42 may be used. In some embodiments, a of hair growth in individuals, such as humans, e.g., by combination of TGF-B mimics is used. promoting proliferation of hair follicle stem cells. In one 0066. In some embodiments, the TGF-B mimic contains embodiment, the method comprises administering to the a peptide sequence that is at least about 70%, 80%, 90%, or skin in the area in which hair growth is desired an amount 95% identical to one of SEQ ID NOS: 1-42, or SEQ ID of TGF-B mimic sufficient to increase hair follicle size NOS: 1, 2, 3, 1, 13, or 14, or SEQ ID NO: 1. and/or the rate of hair growth in the animal, e.g., human. Typically, the composition will be administered topically as 0067. The TGF-B mimics may be administered in any a cream or lotion, and will be applied on a daily basis until cosmetically acceptable carrier, as described in more detail hair growth is observed and for a time thereafter sufficient to below. In embodiments of methods of the invention, the maintain the desired amount of hair growth. concentration of TGF-B mimic used may be more than about 0062 Undesired pigmentation includes pigmentation 0.00001, 0.00005, 0.0001, 0.001, 0.01, 0.1, 1, or 5%. In over an area of the body that is different than the pigmen some embodiments, the concentration of the TGF-B mimic tation desired by the individual. Undesired pigmentation can is more than about 0.00005%. The concentration of TGF-f be the result of, e.g., photoaging, reaction to inflammation, mimic may be less than about 0.0001, 0.001, 0.001, 0.01, or reation to trauma Such as Surgical or accidental skin 0.1. 1, 5, or 10% (all concentration percentages given herein breakage, and the like. Undesired pigmentation includes are w/w unless otherwise indicated). In some embodiments, altered or undesired pigmentation over Small areas such as the TGF-B mimic is used at a concentration of about freckles, as well as altered or undesired pigmentation over 0.00001% to about 1%; or about 0.00001% to about 0.1%: larger areas, such as, for example, uneven pigmentation or or about 0.0001% to about 0.01%; or about 0.0005% to larger areas of undesired pigmentation. about 0.005%; or about 0.0005% to about 0.002%; or about 0.001%. In some embodiments, lower and higher concen 0063. Further cosmetic uses of the methods of the inven tion include tissue augmentation through, generally, topical trations, e.g. 0.00001-0.001%, and 0.001-0.01% are contem application, such as for lip enhancement. By “augment' is plated. In some embodiments, even higher concentrations meant to include giving the appearance of greater fullness, may be warranted, e.g. 0.01%-0.1% or 0.1%-1%, or 0.1%- generally through an increase in the tissue of the skin or 5%, or 0.1%-10%. underlying tissue. Any Suitable skin area may be selected for 0068 Skin coverage may also be described in terms of augmentation by the methods of the invention. total ng of TGF-?3 mimic/cm of skin; in these terms, a US 2006/0293227 A1 Dec. 28, 2006 typical coverage per administration would be more than 0072 Some embodiments of cosmetic treatment of skin about 3, 6, 60, 600, 6000, 60,000, or 600,000 ng/cm of skin; employ topical administration of a lotion, in some embodi less than about 900,000, 600,000, 60,000, 6000, 600, 60, or ments a mixture of emulsifying lanolin alcohols, waxes, and 6 ng/cm of skin; or about 6 ng/cm to about 600 ng/cm of oils (e.g., EUCERINTM) or a mixture of petrolatum or skin; or about 60 ng/cm of skin. mineral oil, a quaternary ammonium compound, a fatty alcohol, and a fatty ester emollient (e.g., CURELTM, see U.S. 0069. The methods of the invention typically utilize Pat. No. 4.389.418), as described below, comprising a topical administration, which may be by any Suitable means TGF-B mimic. In some embodiments, the TGF-B mimic that brings the TGF-B mimic and, optionally, other cosmetic contains a peptide sequence that is at least about 70%, 80%, or dermatological agents, in contact with the Surface of the 90%, or 95% identical to one of SEQID NOS: 1-42, or SEQ skin, including application as a gel, lotion, cream, liposomal ID NOS: 1, 2, 3, 1, 13, or 14, or SEQ ID NO: 1. In some preparation or the like, with or without occlusion, or appli embodiments, the methods utilize cytomodulin (SEQ ID cation as a plaster, patch, mask, glove, or similar device for NO: 1). The concentration of the TGF-B mimic (e.g., cyto extended contact with an affected area of skin. For modu modulin) can be more than about 0.00005%, 0.0001%; lation of cosmetic conditions or to produce a desired cos 0.001%; 0.01%, 0.1%; 1%, or 10%; the TGF-1 mimic can be metic effect, the frequency and duration of administration of at a concentration of less than about 15%, 10%, 1%, 0.1%, a formulation comprising a TGF-3 mimic is dependent on 0.01%; 0.001%; or 0.0001%; in some embodiments, about factors including the nature of the formulation (e.g., con 0.0001% to about 0.01%, in some embodiments about centration, presence or lack of other cosmetic or dermato 0.001%. The lotion containing the TGF-B mimic (e.g., logical agents, vehicle type), the severity and extent of the cytomodulin) is applied at a frequency of once to three times condition, and in some cases the judgment of a skin care per day, in some embodiments once per day, until the desired professional, e.g., a health care professional Such as a result, e.g., reduction or elimination of wrinkling, sagging, dermatologist, or a cosmetologist. and the like, is observed, followed by topical application once to three times per week, in Some embodiments once per 0070 Topical application may be more than about once, week, thereafter. twice, three times, four times, five times, or six times per week, or more than once, twice, three times, four times, five 0073) If the TGF-B mimic is used in combination with times, or six times per day. Frequency of application may be another skin care method or composition, any Suitable less than about twice, three times, four times, five times, or combination of the TGF-B mimic and the additional method six times per week, or less than about once, twice, three or composition may be used. Thus, for example, if use of a times, four times, five times, or six times per day. Some TGF-B mimic is in combination with another cosmetic or embodiments of the invention provide a method for cos dermatological agent, the two may be administered simul metic treatment of the skin of an individual by topical taneously, consecutively, in overlapping durations, in simi administration of a an effective amount of a TGF-B mimic. lar, the same, or different frequencies, etc. In some cases a In some embodiments, the formulation is administered an composition will be used that contains a TGF-B mimic in average of about once per day; in some embodiments, the combination with one or more other cosmetic or dermato formulation is administered an average of about once or logical agents. twice per day; in Some embodiments, the formulation is 0074. Other dermatological or cosmetic agents that may administered an average of about once to three times per be used in methods of the invention are described in more day; in Some embodiments, the formulation is administered detail below. Dosages, routes of administration, administra an average of more than about three times per day. In one tion regimes, and the like for these agents are well-known in embodiment, the formulation is administered an average of the art. about twice per day, typically in the morning upon rising and in the evening before retiring. Topical administration may be 0075 III. Compositions without a covering. Alternatively, topical administration 0076 Another aspect of the present invention relates to may include the use of a covering over the formulation, cosmetic compositions containing a TGF-B mimic. In some which may be occlusive or non-occlusive. For example, embodiments, a combination of TGF-3 mimics is used. In administration in the evening before retiring may include some embodiments the TGF-B mimic(s) is in combination covering the administered area with an occlusive or non with other cosmetic or dermatological ingredients, as occlusive covering, which may remain in place during sleep. described herein. Such compositions are used to modulate 0071. The duration of treatment generally will depend on cosmetic conditions or to produce desirable cosmetic results. the response of the skin to cosmetic treatment. Treatment 0077. As the cosmetic compositions are for topical use, may continue at the discretion of the individual being they need not be sterile; however, if sterility is desired it is treated. In some cases, administration or application of a readily accomplished by filtration through sterile filtration formulation containing a TGF-B mimic may be more fre quent at the beginning of treatment and less frequent as (0.22 micron) membranes, or by other art-accepted means. treatment continues and the condition is ameliorated or the 0078. In cosmetic compositions of the invention, the desired effect is achieved. In some cases, treatment may TGF-B mimic may also be formulated as a cosmetically continue indefinitely in order to maintain a condition in acceptable salt or cosmetically acceptable ester or amide. As abeyance or in an improved State, to delay onset of a used herein, the term “TGF-B mimic’ includes all cosmeti cosmetic condition (e.g., skin aging), or to slow the pro cally acceptable salts or cosmetically acceptable esters or gression of a cosmetic condition. These modifications of amides thereof. The term "cosmetically acceptable salt” or frequency and duration are easily accomplished by the "cosmetically acceptable ester or amide' means those salts, individual being treated. esters, or amides that retain the cosmetic effectiveness and US 2006/0293227 A1 Dec. 28, 2006 properties of the compounds used in the present invention. 0084 Compositions known in the art, preferably For example, a cosmetically acceptable salt does not inter hypoallergic and pH controlled are especially preferred for fere with the cosmetically acceptable effect of a TGF-B topical administration, and include toilet waters, packs, mimic in modulating an age-related condition Such as wrin lotions, skin milks or milky lotions. The preparations con kling. tain, besides the TGF-B mimic and, optionally, other active 0079 TGF-B mimics form cosmetically acceptable salts ingredients, components usually employed in Such prepara with organic and inorganic acids and can be administered in tions. Examples of Such components are oils, fats, waxes, salt form or the TGF-B mimic can be amidated. Examples of Surfactants, humectants, thickening agents, antioxidants, suitable acids for the formation of cosmetically acceptable Viscosity stabilizers, chelating agents, buffers, preservatives, salts are hydrochloric, Sulfuric, phosphoric, acetic, benzoic, perfumes, dyestuffs, lower alkanols, and the like. citric, malonic, Salicylic, malic, fumaric, succinic, tartaric, 0085 Examples of oils include fats and oils such as olive lactic, gluconic, ascorbic, maleic, benzene-Sulfonic, meth oil and hydrogenated oils; waxes such as beeswax and ane- and ethanesulfonic, hydroxymethane- and hydroxy lanolin; hydrocarbons such as liquid paraffin, ceresin, and ethane-Sulfonic. squalene; fatty acids such as Stearic acid and oleic acid; alcohols such as cetyl alcohol, Stearyl alcohol, lanolin alco 0080 Salts may also be formed with suitable organic hol, and hexadecanol; and esters such as isopropyl myristate, cosmetically acceptable base addition salts. These organic isopropyl palmitate and butyl Stearate. As examples of bases form a class whose limits are readily understood by Surfactants there may be cited anionic Surfactants such as those skilled in the art. Merely for purposes of illustration, Sodium Stearite, sodium cetylsulfate, polyoxyethylene lau the class may be said to include mono-, di-, and trialky rylether phosphate, sodium N-acyl glutamate; cationic Sur lamines, such as methylamine, dimethylamine, and triethy factants such as Stearyldimethylbenzylammonium chloride lamine; mono-, di-, or trihydroxyalkylamines such as and Stearyltrimethylammonium chloride; ampholytic Surfac mono-, di-, and triethanolamine; amino acids such as argi tants such as alkylaminoethylglycine hydrochloride solu nine, and lysine; guanidine; N-methyl-glucosamine; N-me tions and lecithin; and nonionic Surfactants such as glycerin thyl-glucamine; L-glutamine; N-methylpiperazine; morpho monostearate, Sorbitan monostearate. Sucrose fatty acid line; ethylenediamine; N-benzylphenethylamine: esters, propylene glycol monostearate, polyoxyethylene tris(hydroxymethyl)aminomethane; and the like. (See, for oleylether, polyethylene glycol monostearate, polyoxyeth example, “Pharmaceutical Salts.” J. Pharm. Sci., 66(1), 1-19 ylene Sorbitan monopalmitate, polyoxyethylene coconut (1977), which are acceptable as cosmetically acceptable fatty acid monoethanolamide, polyoxypropylene glycol (e.g. salts as well.) the materials sold under the trademark “Pluronic'), poly 0081. If desired, further cosmetic or dermatological oxyethylene castor oil, and polyoxyethylene lanolin. ingredients may be incorporated in the formulations. The Examples of humectants include glycerin, 1.3-butylene gly nature of the other ingredient(s) will depend on the cosmetic col, and propylene glycol; examples of lower alcohols condition to be modulated and/or cosmetic result desired. include ethanol and isopropanol; examples of thickening These are described more fully below. agents include Xanthan gum, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyethylene glycol and 0082 The TGF-3 mimic may be used neat (e.g., with an Sodium carboxymethyl cellulose; examples of antioxidants occlusive dressing so that the TGF-3 mimic is dissolved or include butylated hydroxytoluene, butylated hydroxyani dispersed in perspiration at the site), but generally is pre sole, propyl gallate, citric acid and ethoxyquin; examples of pared in a vehicle Suitable for topical administration. Com chelating agents include disodium edetate and ethanehy positions of the invention include TGF-B mimic in a vehicle droxy diphosphate; examples of buffers include citric acid, Suitable for topical administration. Sodium citrate, boric acid, borax, and disodium hydrogen 0083) Numerous vehicles for topical application of cos phosphate; and examples of preservatives are methyl metic compositions are known in the art. See, e.g., Reming parahydroxybenzoate, ethyl parahydroxybenzoate, dehy ton's Pharmaceutical Sciences, Gennaro, A R, ed., 20" droacetic acid, Salicylic acid and benzoic acid. These Sub edition, 2000: Williams and Wilkins PA, USA. All compo stances are merely exemplary, and those of skill in the art sitions usually employed for topically administering cos will recognize that other substances may be substituted with metic compositions may be used, e.g., creams, lotions, gels, no loss of functionality. dressings, shampoos, tinctures, pastes, ointments, salves, 0086 For preparing compositions for topical administra powders, liquid or semiliquid formulation, patches, liposo tion, the concentration of TGF-B mimic may be more than mal preparations, and the like. Application of said compo about 0.00001, 0.00005, 0.0001, 0.001, 0.01, 0.1, 1, or 5%. sitions may, if appropriate, be by aerosol e.g. with a pro In some embodiments, the concentration of the TGF-B pellant Such as nitrogen carbon dioxide, a freon, or without mimic is more than about 0.00005%. The concentration of a propellant such as a pump spray, drops, lotions, or a TGF-B mimic may be less than about 0.0001, 0.001, 0.001, semisolid Such as a thickened composition which can be 0.01, 0.1. 1, 5, or 10% (all concentration percentages given applied by a Swab. In particular compositions, semisolid herein are w/w unless otherwise indicated). In some embodi compositions such as Salves, creams, lotions, pastes, gels, ments, the TGF-B mimic is incorporated at a concentration ointments and the like will conveniently be used. The TGF-B of about 0.00001% to about 1%; or about 0.00001% to about mimic may optionally be dissolved in a small amount (e.g., 0.1%; or about 0.0001% to about 0.01%; or about 0.0005% less than 100, less than 10, or less than 1 ug/ml TGF-B to about 0.005%; or about 0.0005% to about 0.002%; or mimic; in some embodiments, less than 1 ug/ml TGF-B about 0.001%. In some embodiments, lower and higher mimic) of an appropriate solvent, such as ethanol or DMSO, concentrations, e.g. 0.00001-0.001%, and 0.001-0.01% are before dispersion in the vehicle; however, this is not contemplated. In some embodiments, even higher concen required. trations may be warranted, e.g. 0.01%-0.1% or 0.1%-1%, or US 2006/0293227 A1 Dec. 28, 2006

0.1%-5%, or 0.1%-10%. In some embodiments employing TGF-B mimic in a lotion comprising a mixture of petrolatum ointments, lotions, or creams, the carrier for example, can or mineral oil, a quaternary ammonium compound, a fatty contain 1 to 20%, in particular 5 to 15% of a humectant, 0.1 alcohol, and a fatty ester emollient (e.g., CURELTM Fra to 10% in particular from 0.5 to 5% of a thickener and water; grance Free Daily Moisturizing Lotion) at a concentration of or said carrier may consist of 70 to 99%, in particular 20 to about 0.0001% to about 0.01%. Some embodiments com 95% of a surfactant, and 0 to 20%, in particular 2.5 to 15% prise cytomodulin (SEQ ID NO: 1) in a lotion comprising a of a fat; or 80 to 99.9% in particular 90 to 99% of a mixture of petrolatum or mineral oil, a quaternary ammo thickener; or 5 to 15% of a surfactant, 2-15% of a humectant, nium compound, a fatty alcohol, and a fatty ester emollient 0 to 80% of an oil, very small (<2%) amounts of preserva (e.g., CURELTM Fragrance Free Daily Moisturizing Lotion) tive, coloring agent and/or perfume, and water. In a toilet at a concentration of about 0.001%. water, the carrier for example consists of 2 to 10% of a lower 0092. In addition, a TGF-B mimic and, optionally, other alcohol, 0.1 to 10% or in particular 0.5 to 1% of a surfactant, active ingredients, may be formulated in liposome-contain 1 to 20%, in particular 3 to 7% of a humectant, 0 to 5% of ing compositions. Liposomes are artificial vesicles formed a buffer, water and small amounts (<2%) of preservative, by amphipathic molecules such as polar lipids, for example, dyestuff and/or perfume. In a skin milk, the carrier typically phosphatidylcholines, ethanolamines and serines, sphingo consists of 10-50% of oil, 1 to 10% of surfactant, 50-80% of myelins, cardiolipins, plasmalogens, phosphatidic acids and water and 0 to 3% of preservative and/or perfume. cerebrosides. Liposomes are formed when Suitable amphi 0087. In some embodiments, compositions of the inven pathic molecules are allowed to Swell in water or aqueous tion contain a TGF-B mimic that contains a peptide sequence Solutions to form liquid crystals usually of multilayer struc that is at least about 70%, 80%, 90%, or 95% identical to one ture comprised of many bilayers separated from each other of SEQID NOS: 1-42, or SEQID NOS: 1, 2, 3, 1, 13, or 14, by aqueous material (also referred to as coarse liposomes). or SEQ ID NO: 1. In some embodiments, the compositions Another type of liposome known to be consisting of a single contain cytomodulin (SEQ ID NO: 1). bilayer encapsulating aqueous material is referred to as a unilamellar vesicle. If water-soluble materials are included 0088. In some embodiments, the lotion in which one or in the aqueous phase during the Swelling of the lipids they more TGF-B mimics, optionally with other active ingredi become entrapped in the aqueous layer between the lipid ents, is (dissolved, mixed, or Suspended) in a mixture of bilayers. emulsifying lanolin alcohols, waxes, and oils (e.g., EUC ERINTM Lotion) or a mixture of petrolatum or mineral oil, 0093. The incorporation of TGF-B mimics into liposomal a quaternary ammonium compound, a fatty alcohol, and a preparations suitable for topical application can be achieved fatty ester emollient (e.g., CURELTM Lotion), or lotions that by a number of methods. With respect to liposomal prepa are substantially similar in composition. rations, any known methods for preparing liposomes for treatment of a condition may be used. See, for example, 0089 EUCERINTM Dry Skin Therapy Original Moistur Bangham et al., J. Mol. Biol, 23: 238-252 (1965) and Szoka izing Lotion comprises water, mineral oil, isopropyl et al., Proc. Natl Acad. Sci. 75: 4194-4198 (1978). Ligands myristate, PEG-40 sorbitan peroleate, glyceryl lanolate, may also be attached to the liposomes to direct these Sorbitol, propylene glycol, cetyl palmitate, magnesium Sul compositions to particular sites of action. fate, aluminum stearate, lanolin alcohol, BHT methylchlor oisothiazolinone, and methylisothiazolinone. CURELTM 0094 Liposomes containing a TGF-B mimic and, option Fragrance Free Daily Moisturizing Lotion comprises water, ally, other ingredients can be employed directly or they can glycerin, distearyldimonium chloride, petrolatum, isopropyl be employed in a suitable pharmaceutically acceptable car palmitate, cetyl alcohol, dimethicone, sodium chloride, rier for topical administration. The viscosity of the lipo methylparaben, and propylparaben. Somes can be increased by the addition of one or more Suitable thickening agents such as, for example Xanthan 0090 Some embodiments of compositions of the inven gum, hydroxypropyl cellulose, hydroxypropyl methyl cel tion comprise a TGF-B mimic in a lotion comprising a lulose and mixtures thereof. The aqueous component may mixture of emulsifying lanolin alcohols, waxes, and oils consist of water alone or it may contain electrolytes, buff (e.g., EUCERINTM Dry Skin Therapy Original Moisturizing ered systems and other ingredients, such as, for example, Lotion) at a concentration greater than about 0.00005%. preservatives. Suitable electrolytes which can be employed Some embodiments of compositions of the invention com include metal salts such as alkali metal and alkaline earth prise a TGF-3 mimic in a lotion comprising a mixture of metal salts. Exemplary metal salts are calcium chloride, emulsifying lanolin alcohols, waxes, and oils (e.g., EUC Sodium chloride and potassium chloride. The concentration ERINTM Dry Skin Therapy Original Moisturizing Lotion) at of the electrolyte may vary from Zero to 260 mM, preferably a concentration of about 0.0001% to about 0.01%. Some from 5 mM to 160 mM. The aqueous component is placed embodiments comprise cytomodulin (SEQ ID NO: 1) in a in a suitable vessel which can be adapted to effect homog lotion comprising a mixture of emulsifying lanolin alcohols, enization by effecting great turbulence during the injection waxes, and oils (e.g., EUCERINTM Dry Skin Therapy Origi of the organic component. Homogenization of the two nal Moisturizing Lotion) at a concentration of about 0.001%. components can be accomplished within the vessel, or, 0.091 Some embodiments of compositions of the inven alternatively, the aqueous and organic components may be tion comprise a TGF-B mimic in a lotion comprising a injected separately into a mixing means which is located mixture of petrolatum or mineral oil, a quaternary ammo outside the vessel. In the latter case, the liposomes are nium compound, a fatty alcohol, and a fatty ester emollient formed in the mixing means and then transferred to another (e.g., CURELTM Fragrance Free Daily Moisturizing Lotion) vessel for collection purpose. at a concentration greater than about 0.00005%. Some 0095 The organic component consists of a suitable non embodiments of compositions of the invention comprise a toxic, cosmetically acceptable solvent Such as, for example US 2006/0293227 A1 Dec. 28, 2006 ethanol, glycerol, propylene glycol and polyethylene glycol, variety of agents, often in combination, that prevent or treat and a suitable phospholipid which is soluble in the solvent. wrinkling through a variety of actions. Many approaches are Suitable phospholipids which can be employed include taken to reduce the appearance of facial and other wrinkles lecithin, phosphatidylcholine, phosphatidylserine, phos based on the understanding of the molecular basis of wrinkle phatidylethanolamine, phosphatidylinositol, lysophosphati formation. Such treatments include cosmetic products, drug dylcholine and phosphatidylglycerol, for example. Other therapy and Surgical procedures. For example, many cos lipophilic additives may be employed in order to selectively metic products contain hydroxy acids, which may stimulate modify the characteristics of the liposomes. Examples of collagen synthesis. Another common treatment utilizes ret Such other additives include Stearylamine, phosphatidic inol, retinoic acid, retinol palmitate, a derivative of vitamin acid, tocopherol, cholesterol and lanolin extracts. In addi A, (or prescribed versions, Retin-A and Renova) which may tion, other ingredients which can prevent oxidation of the directly or indirectly stimulate collagen production or retard phospholipids may be added to the organic component. collagen degradation. Bicyclic aromatic compounds with Examples of Such other ingredients include tocopherol, retinoid-type activity, which are useful in particular in butylated hydroxyanisole, butylated hydroxytoluene, ascor preventing or treating various keratinization disorders, are byl palmitate and ascorbyl oleate. Preservatives such a described in EP 679 630. These compounds are particularly benzoic acid, methyl paraben and propyl paraben may also active for repairing or combating chronological or actinic be added. ageing of the skin, for example such as in anti-wrinkle products. Antioxidants such as vitamin C and E and coen 0.096 Although TGF-B mimic are generally capable of Zyme Q-10 are believed to counteract free radicals, which penetrating cell membranes and reaching the deep layers of damage cells and cause aging and have been used in skin, it may be useful in some embodiments to also include treatments of wrinkles. Recently, the FDA approved cos a penetration enhancer in the formulations of the invention. metic use of Botox (an extremely purified form of botulinum A penetration enhancer is a substance that improves cuta toxin) to treat glabella frown lines. neous penetration of a bioactive Substance. Suitable pen etration enhancers include, for example, dimethyl Sulfoxide 0.100 Thus cosmetic or dermatological agents that (DMSO), DMSO-like compounds, ethanolic compounds, complement cosmetic treatment of skin with the methods or pyroglutamic acid esters and other solvents or compounds compositions of the invention include, alone or in combi known to those skilled in the pharmaceutical art which nation, the bicyclic aromatic compounds defined above, facilitate dermal penetration of the drugs or chemicals other compounds which have retinoid-type activity, free chosen for the pharmaceutical composition. Other penetra radical scavengers, hydroxy or keto acids or derivatives tion enhancers include amphiphiles such as L-amino acids, thereof. anionic Surfactants, cationic Surfactants, amphoteric Surfac tants, nonionic Surfactants, fatty acids and alcohols. Addi 0101 The term “free-radical scavenger” refers to, for tional penetration enhancers are disclosed in Remington: example, C-tocopherol, Superoxide dismutase, ubiquinol The Science and Practice of Pharmacy, 19th Edition (1995) (e.g., coenzyme Q10) or certain metal-chelating agents. on page 1583. The penetration enhancer chosen and the Hydroxy acids include, e.g., alpha-hydroxy acids such as relative proportion of the penetration enhancer with respect lactic acid and glycolic acid or beta-hydroxy acids such as to the active drugs or chemicals depends on the desired rate salicylic acid and salicylic acid derivatives such as the of delivery of the drugs or chemicals into the skin, which in octanoyl derivative; other hydroxy acids and keto acids turn depends on the condition being treated and the outcome include malic, citric, mandelic, tartaric or glyceric acids or sought. More specifically, the type and amount of enhancer the salts, amides or esters thereof. is chosen so that a sufficiently high concentration of active 0102). Other anti-wrinkling agents and anti-skin aging drugs or chemicals is attained in the skin to treat the agents useful in the invention include Sulfur-containing D condition within the time period considered desirable. and L amino acids and their derivatives and salts, particu 0097 Apart from the above-described compositions, use larly the N-acetyl derivatives, a preferred example of which may be made of covers, e.g. plasters, bandages, dressings, is N-acetyl-L-cysteine; thiols, e.g. ethane thiol: fat-soluble gauze pads, patches and the like, containing an appropriate Vitamins, ascorbyl palmitate, ceramides, pseudoceramides amount of a TGF-B mimic and, optionally, other ingredients. (e.g., pseudoceramides described in U.S. Pat. Nos. 5,198. In some cases use may be made of plasters, bandages, 210; 4,778,823: 4,985,547; 5,175,321, all of which are dressings, gauze pads, patches and the like which have been incorporated by reference herein), phospholipids (e.g., dis impregnated with a topical formulation containing the thera tearoyl lecithin phospholipid), fatty acids, fatty alcohols, cholesterol, plant sterols, phytic acid, lipoic acid; lysophos peutic formulation. phatidic acid, and skin peel agents (e.g., phenol and the like), 0098. Additional cosmetic and dermatological agents. and mixtures thereof. Preferred fatty acids or alcohols are Other cosmetic or dermatological agents may be included in those that have straight or branched alkyl chains containing methods and formulations of the invention. A "cosmetic or 12-20 carbon atoms. A particularly preferred fatty acid is dermatological agent as used herein, includes any Sub linoleic acid since linoleic acid assists in the absorption of stance whose administration for treatment of a cosmetic ultraviolet light and furthermore is a vital component of the condition or to achieve a desired cosmetic effect, results in natural skin lipids. Other non-limiting examples of Suitable an status of the condition that is better than the status that anti-wrinkle actives for use herein are described in U.S. Pat. would exist without the use of the cosmetic or dermatologi No. 6,217,888, which description is incorporated herein by cal agent. reference. 0099 Anti-wrinkling agents are one form of cosmetic or 0.103 Compositions for cosmetic treatment of skin may dermatological agents. Anti-skin wrinkling agents include a further include Sunscreens to lower skin's exposure to US 2006/0293227 A1 Dec. 28, 2006

harmful UV rays. Sunscreens include those materials com above references, as well as Textbook of Dermatology, monly employed to absorb or block ultraviolet light. Illus Champion, Burton, Burns, and Bretnach, eds., Blackwell trative compounds are the derivatives of PABA, cinnamate Publishing, 1998. and derivatives of salicylate (other than ferulyl salicylate). For example, octyl methoxycinnamate and 2-hydroxy-4- 01.09) IV. Kits of the Invention methoxybenzophenone (also known as oxybenzone) can be 0110. In still another aspect, the present invention pro used. Octyl methoxycinnamate and 2-hydroxy-4-methoxy vides kits for the cosmetic treatment of skin or to produce a benzophenone are commercially available under the trade desired cosmetic result. These kits comprise a TGF-B mimic marks, PARSOL MCX and BENZOPHENONE-3, respec or TGF-B mimic composition or compositions described tively. DermaScreen may also be used. herein, in a container or containers which are held in Suitable packaging. In some embodiments the kits further contain 0104 Many other sunscreens are known to those of skill instructions teaching the use of the kit according to the in the art. In some embodiments, Sunscreens are FDA various methods and approaches described herein. Such kits approved or approved for use in the European Union. For may also include information, such as Scientific literature example, FDA-approved Sunscreens may be used, singly or, references, package insert materials, cosmetic trial results, preferably, in combination. See, e.g., U.S. Pat. Nos. 5,169, and/or summaries of these and the like, which indicate or 624; 5,543,136; 5,849,273; 5,904,917; 6,224,852: 6,217, establish the activities and/or advantages of the composition. 852; and Segarinet al., chapter Vil, pages 189 of Cosmetics Such information may be based on the results of various Science and Technology, and Final Over-the-Counter Drug studies, for example, studies using experimental animals Products Monograph on Sunscreens (Federal Register, involving in vivo models and studies based on human 1999:64:27666-27963), all of which are incorporated herein cosmetic or clinical trials. Kits described herein can be by reference. The exact amount of Sunscreen employed provided, marketed and/or promoted to health care providers in-the compositions can vary depending upon the degree of (e.g., dermatologists and other physicians), skin care appear protection desired from the sun's UV radiation. ance care providers, including cosmetologists, hair stylists, 0105 Cosmetic compositions of the invention may fur and the like. Kits for cosmetic use may also be provided, ther include anti-acne agents. Anti-acne agents include ben marketed and/or promoted directly to consumers. Kits may Zoyl peroxide, antibiotics, e.g., erythromycin, clindamycin be marketed in spas and retail outlets. phosphate, 5,7-dichloro-8-hydroxyquinoline, resorcinol, resorcinol acetate, salicylic acid, azaleic acid, long chain EXAMPLES dicarboxylic acids, Sulfur, Zinc, retinoids, antiandrogens, Example 1 and various natural agents such as those derived from green tea, tea tree oil, and mixtures thereof. Other non-limiting Inhibition of DNA Synthesis of MV-1-Lu Mink examples of Suitable anti-acne agents for use herein are Lung Epithelial Cells described in U.S. Pat. No. 5,607,980, which description is 0111. The effect of TGF-B and cytomodulin were evalu incorporated herein by reference. ated by determining the rate of "Hithymidine incorporation into total acid-insoluble DNA and cell number. See gener 0106 Other cosmetic and dermatological agents include ally, Sampath et al., J. Biol. Chem., 267, pp. 20352-20362 anticellulite agents. Anticellulite agents include isobutylm (1992). DNA synthesis rates were determined in triplicate ethylxanthine, caffeine, theophylline, theobromine, amino cultures after 24 hour treatment with various concentrations phylline, yohimbine, and mixtures thereof. (10 M to 10M) of either TGF-3 or cytomodulin (which 0107 Yet other cosmetic or dermatological agents that was synthesized by the Merrifield method) by adding me complement cosmetic treatment of skin include C-inter thyl-Hlthymidine (2uCi/ml, 80 Ci/mmol) for 6 hours before feron, estradiol; progesterone; pregnanalone; methylSola the termination of the culture. Incorporation was terminated nomethane (MSM); copper peptide (copper extract); plank by aspiration of the medium, and after washing three times ton extract (phytosome); broparoestrol; estrone; with phosphate-buffered saline, the trichloroacetic acid adrostenedione; androstanediols; etc. (10%)-precipitated radioactive DNA was extracted with 1.0% (w/v) sodium dodecyl sulfate, 0.1 M NaOH and 0108. The compositions of the present invention may quantitated by liquid Scintillation counting. For cell number contain a wide range of additional components. The CTFA determination, 1x10 cells were plated in flasks in MEM Cosmetic Ingredient Handbook, Seventh Edition, 1997 and containing 10% FBS, and after 24 hours, the growth medium the Eighth Edition, 2000, which are incorporated by refer was replaced with serum-free medium containing various ence herein in their entirety, describes a wide variety of conceptions of TGF-B and cytomodulin. Triplicate cultures ingredients commonly used in skin care compositions, were harvested every 24 hours for the duration of 7 days, and which are suitable for use in the compositions of the present the cell number was determined by counting cells released invention. Other topically-applied compounds are listed in by trypsin digestion in a fixed volume hemacytometer. Remington’s Pharmaceutical Sciences, 20th Ed., Lippincott Williams & Witkins, Baltimore, Md. (2000) (hereinafter 0.112. The growth inhibition curve for cytomodulin were Remington's), U.S. Pharmacopeia and National Formulary, similar to that observed for TGF-B at the same concentration The United States Pharmacopeial Convention, Inc., Rock range. ville, Md. and Physicians Desk Reference, Medical Eco Example 2 nomics Co., Inc., Oradell, N.J. incorporated herein by ref erence. The concentration of the other active ingredient in Growth and Colony Formation by NRK-49 F formulations provided by the invention is that which pro Fibroblasts in Soft Agar vides an effective amount of the other active ingredient; 0113. The original assay for TGF-B, the ability to pro these concentrations are well-known in the art. See, e.g., the mote anchorage independent growth of normal fibroblasts is US 2006/0293227 A1 Dec. 28, 2006

still one of the hallmarks of TGF-B activity. NRK-49 F Moreover, as with cytomodulin, L1 and L2 also increased fibroblasts were grown at 37°C. in DEM supplemented with the expression of type I collagen, TGF-3 and alkaline 10% fetal calf serum. The experiments were performed with phosphatase in HOS cells. culture medium, 10 ng/mg epidermal growth factor (EGF), 0123. Further analogs of SEQ ID NO:2 and SEQ ID and 10 ng/ml platelet-derived growth factor (PDGF); how NO:3 were then made: ever, unlike TGF-B, which does not induce colony formation in the absence of these factors (see, for example, Massagu, J. Biol. Chem., 259, pp.9756-9761 (1984)), cytomodulin did - (Aib)-A-E-A-K (SEQ ID NO: 4) induce colony formation without these two growth factors. To this, either 100 nM TGF-3 (positive control) or 100 nM L-I- (Nme-A)-E-A-K (SEQ ID NO:5) cytomodulin was added. NRK49 F fibroblasts (5x10 cells/ mi) were mixed with 0.3% agar were plated on the bottom - (Abu)-A-E-A-K (SEQ ID NO: 6) of 35 mm culture dishes. Colony formation was observed G-G-Q-I-A-N-I (SEQ ID NO: 7) starting on day 3 of culture. E-G-I-A-G-K (SEQ ID NO: 8) 0114. As expected no colonies were formed in those cultures containing only the basic medium. Also, as --A-D-A-K (SEQ ID NO: 9) expected, colonies with TGF-B grew colonies. Surprisingly, -I-A-N-A-K (SEQ ID NO: 10) the cytomodulin cultures also formed colonies to approxi mately the same extent as the TGF-B cultures. The growth --A-E-A-A. (SEQ ID NO: 11) characteristics of the colonies over time were similar between TGF-B and cytomodulin cultures. L-I-A-Q-A-K (SEQ ID NO: 12) --A-G-G-E (SEQ ID NO: 13) Example 3 --A-G-E-G (SEQ ID NO: 14) Further TGF-B Mimics A-N-W-A.-E.-K. (SEQ ID NO: 15) 0115 FIGS. 1A and 1B show the atomic coordinates of --A-K-G-K (SEQ ID NO: 16) the bioactive structure of cytomodulin (atoms 1-101). Thus, the structure represented by FIGS. 1A and 1B, describes 0.124 Of the non-standard amino acids, Aib is C.-amino one of the possible structures consistent with TGF-B activity isobutyric acid, Nme-Ala is N-methyl alanine and Abu is 0116. Using the three dimensional structure of cytomodu C.-aminobutyric acid. lin (FIGS. 1A and 1B) as a guide, cytomodulin analogs 0.125 SEQID NOs:4-6, which are minor variants of SEQ were designed and tested for TGF-B activity. An exemplary ID NO:2, mimicked the biological activities of TGF-B and general formula to produce a peptide with atomic coordi cytomodulin as shown by the inhibition of the proliferation nates substantially the same as that of FIGS. 1A and 1B, of MV-1-Lu epithelial cells and increased expression of which was used in the construction of these two cytomodu collagen I and TGF-B in HOS cells. Sample thymidine lin analogs, was AA-AA-AA. . . . -AA incorporation data for SEQID NOs:4-6 are shown in Table 0.117 (1) a hydrophobic or neutral amino acid at posi 1. tion i; TABLE 1. 0118 (2) a branched hydrophobic at position i+1; Inhibition of Incorporation of H-thymidine inMV-1 Lucells 0119 (3) a small aliphatic at position i+2, where posi in the Presence of Test Peptides tions i-1 and i--2 together form the U-bend (B-bend) Control H-Radioactivity, 10.sup.3dpm structure; and (No peptides added) (% Inhibition) 3.57 (-) 0120 (4) a side-chain possessing a hydrogen bond (IV) LAib AEAK (SEQ ID NO: 4) acceptor shortly thereafter (at position AA) 1 mM 2.60 (27%) 5 nM 2.31 (35%) 0121 Initially, two cytomodulin analogs, L-I-A-E-A-K SOM 1.94 (46%) (SEQ ID NO: 2 or L2) and L-I-A-P-E-A (SEQID NO:3 or 100 nM 1.44 (60%) L1) were synthesized and tested. In both peptides, -V-A- was SOOM 1.61 (55%) replaced by -I-A- and the first two N-terminal amino acid (V) LINmeAEAK (SEQ ID NO: 5) sequence of cytomodulin was replaced with leucine. In SEQ 1 nM 1.61 (47%) ID NO:2, glutamic acid is at position i+3 since it is the first 5 nM 1.76 (42%) side-chain after the U (B)-bend structure. In SEQ ID NO:3, SOM 1.60 (41%) 100 nM 1.81 (40%) proline is at position i+3. Since proline does not have a SOOM 1.42 (53%) 'side-chain,” the glutamic acid was placed at position i+4. (VI) LAbu AEAK (SEQ ID NO: 6) 0122 Both cytomodulin analogs, L1 and L2, displayed at 1 nM 2.02 (33%) least as much TGF-B like activity as cytomodulin. They 5 nM 1.97 (35%) promoted the growth of NRK-49F cells in soft agar and inhibited the proliferation of MV-1-Lu cells. They increased the expression of type I collagen and TGF-B and decreased 0126) However, SEQID NOS:7-8 did not display signifi the expression of collagenase in human dermal fibroblasts. cant TGF-B activity and thus, as that term is defined herein, US 2006/0293227 A1 Dec. 28, 2006 18 and by the assays used, would not be defined as TGF-B mimics since, although they possessed an atomic structure -continued substantially the same as that shown in FIGS. 1A and 1B, SEQ ID NO 30: Gly Ile Ala Gly Glin they did not show TGF-B activity. This was not unexpected given the working model; e.g., although a glutamic acid is SEQ ID NO 31 : Glin Gly Ala Ile Ala Glin present in SEQ ID NO:8, it is on the N-terminal side of the SEQ ID NO 32: Phe Gly Ile Ala Gly Phe B-bend and not on the C-terminal side as with cytomodulin, L1, and L2. SEQ ID NO 33: Cys Gly Ile Ala Gly Cys 0127. Inhibition of H-thymidine incorporation results SEQ ID NO 34: Glu Gly Ile Ala Gly Lys for SEQ ID NOs:9-16 are shown in Table 2. The numbers SEQ ID NO 35 : Xaa Ile Ala Ala shown at the various concentration are the ratio of the inhibition rate of the peptide being tested over the inhibition SEQ ID NO 36: Ile Ala Xaa rate of cytomodulin (SEQ ID NO: 1) at the same concen tration. Because cytomodulin inhibits the proliferation of SEQ ID NO 37: Xaa Ile Ala Xaa MV-1-Lu cells at least as much as TGF-B, cytomodulin and SEQ ID NO 38: Ile Ile Xaa Glu Ala Lys not TGF-3 was used as a control. SEQ ID NO 39: Leu Ile Xaa Glu Ala Lys TABLE 2 SEQ ID NO 40 : Lieu. Ile Ala Xaa Ala Lys (Inhib by new peptide? SEQ ID NO 41: Lieu. Ile Ala Pro Xaa Ala Inhib by cytomodulin) Peptide Concentration (nM SEQ ID NO 42: Lieu. Ile Ala Xaa Ala Lys Composition SEQ ID NO 1 10 1OO 1OOO 0.130 Tests for bioactivity were conducted as described LIADAK SEQ ID NO: 9 O45 O.87 1.14 1.25 LIANAK SEQ ID NO: 10 1.70 2.00 1.43 3.05 above for DNA inhibition. In summary, the cytomodulin LIAEAA SEQ ID NO: 11 1.16 1.00 1.17 1.85 analogs listed in Table 3 were all found active as agonists LIAQAK SEQ ID NO: 12 O.90 1.10 O.8O 1.56 and thus, as defined herein, are considered TGF-3 mimics. LIAGGE SEQ ID NO: 13 1.10 1.33 1.45 1.90 LIAGEG SEQ ID NO: 14 O.66 O.85 1.41 1.88 TABLE 3 ANVAEK SEQ ID NO: 15 O.80 1.OO LIAKGK SEQ ID NO: 16 O.65 O.67 SEQ ID SEQUENCE SEQUENCE SYMBOL NO 0128. As illustrated by Table 2, all peptides with Ala-Asn-Wal-Ala-Glu- A-N-W-A-E-N-A 1 sequences represented by SEQ ID NOs:9-16 inhibited at Asn-Ala least some amount of thymidine uptake. Thus, all peptides Leu-Ile-Ala-Pro-Glu- T-I-A-P-E-A 3 with sequences represented by SEQ ID NOs:9-16 are con Ala sidered TGF-B mimics. Leu-Ile-Ala-Glu-Ala- T-I-A-E-A-K 2 0129. Further analogs were constructed (SEQ ID NOS: Lys 17-42): Ile-Aib-Ala-Glu-Ala- I- (Aib)-A-E-A-K 18 Lys SEQ ID NO 17: Gly. Thr Pro Gly Pro Gln Gly Ile Ala Ile- (Ile) - (Nme-Ala) - I- (I) - (NMeA)-E-A-K 38 Gly Glin Arg Gly Val Val Glu-Ala-Lys SEQ ID NO 18: Ile Xaa Ala Glu Ala Lys Leu- (Abu)-Ala-Glu-Ala- L- (Abu)-A-E-A-K 19 Lys SEQ ID NO 19: Lieu Xaa Ala Glu Ala Lys Leu-Ile-Ala-Asn-Ala- -I-A-N-A-K 10 SEQ ID NO 20 : Leu Pro Ala Glu Ala Lys Lys SEQ ID NO 21 : Leu Ile Pro Glu Ala Lys Leu-Ile-Ala-Glu-Ala- --A-E-A-A. 11 Ala SEQ ID NO 22: Lieu. Ile Xaa Glu Ala Lys Leu-Ile-Ala-Lys-Gly- --A-K-G-K 16 SEQ ID NO 23: Leu. Ile Ala Xaa Glu Ala Lys SEQ ID NO 24 : Ile Trp Gly Lieu. Asp Gly Xala Lys Leu-Pro-Ala-Glu-Ala- -P-A-E-A-K 20 Lys SEQ ID NO 25: Trp Ile Ala Leu Glu Gly Xaa Lys Leu-Ile-Pro-Glu-Ala- -T-P-E-A-K 21 SEQ ID NO 26: Gly Pro Glin Gly Ile Ala Gly Glin Arg Lys SEQ ID NO 27: Glin Gly Ile Ala Gly Glin Leu-Ile- (Aib)-Glu-Ala- L-I- (Aib)-E-A-K 22 Lys SEQ ID NO 28: Glin Gly Ile Ala Gly Glin Arg Leu-Ile- (D-Ala)-Glu- L-I- (D-Ala) -E-A-K 39 SEQ ID NO 29: Phe Gly Ile Ala Gly Phe Ala-Lys US 2006/0293227 A1 Dec. 28, 2006

Example 5 TABLE 3-continued

SEQ ID Cosmetic Treatment of Eyelids SEQUENCE SEQUENCE SYMBOL NO 0.134. A female in her 40's, of Asian ethnicity, applied a Leu-Ile-Ala- (D-Glu)- L-I-A-(D-Glu)-A-K 40 lotion comprising cytomodulin (SEQ ID NO: 1) at a con Ala-Lys centration of about 0.001%, to her eyelids twice per week for Leu-Ile-Ala- (Aib)-Glu- L-I-A- (Aib)-E-A 23 2 months. She reported that the effect is like having an eye Ala lift. No adverse effects were noted. Leu-Ile-Ala-Pro- (D- L-I-A-P- (D-Glu)-A 41 Glu)-Ala Example 6 Leu-Ile-Ala- (X. sub. 1) - L-I-A- (X)-A-K 42 Ala-Lys Cosmetic Treatment of Environmental and Chronological Skin Aging Ile-Trp-Gly-Leu-Asp- I-W-G-L-D-G- (bAla) -K 24 Gly-bAla-Lys 0.135 A 51-year old Caucasian male, with very dry, Trp-Ile-Ala-Leu-Glu- W-I-A-L-E-G- (bAla) -K 25 wind-burnt skin, many fine lines, and the beginning of Gly-bAla-Lys wrinkles, applied a lotion comprising cytomodulin (SEQID C.-amino butyric acid NO: 1) at a concentration of about 0.001% on left side of his C.-amino isobutyric acid face only, three days per week. The Subject did not use any (NmeA) = N-methyl alanine X = trans-4-hydroxyproline other other moisturizer of any type on his face. Within 3 weeks there was a a noticeable difference between the two 0131 Similar testing to that described herein, and other sides of the face, with the right side having more fine lines wise available to those of skill in the art to determine appearing much drier and weather-worn than the left TGF-B-like activity, may be applied by those of skill in the (treated) side. The left side looked more supple, plumper, art to peptides that are not included in the above analyses to with fewer lines. The subject stopped application after 3 determine whether or not they are TGF-B mimics. months because of the difference in the appearance of the two side of the face Example 4 0.136. A 48 yr. old Caucasian female applied a lotion Cosmetic Treatment of Skin comprising cytomodulin (SEQID NO: 1) at a concentration 0132 A 61-year old female, of light complexion, applied of about 0.001% on right side of her neck only, daily. The a lotion comprising cytomodulin (SEQ ID NO: 1) at a right side in about three weeks began to look Smoother, more concentration of about 0.001%, in EUCERINTM, to one side Supple, more youthful, and was Smoother to the touch than of her face a at a frequency of 1-2 times per week, typically the left side of the neck. The right side of the neck is at night before sleep, leaving uncovered while leaving the noticeably different than the left side of the neck. No adverse other side of her face without application of the TGF-B effects were noted. The subject accidentally got the lotion in lotion. After the first treatment of topical application of the her eyes, with no apparent problem or irritation. lotion, indices of skin aging, Such as wrinkling (e.g., crows feet), were noticeably ameliorated on the side of the face 0.137. A 59 yr old Caucasian female with noticebly receiving the lotion, compared to the side that did not wrinkled and thickened skin on the neck applied a lotion receive lotion. For example, crows feet became more shal low and the skin and face looked fuller and smoother. Color comprising cytomodulin (SEQID NO: 1) at a concentration and skin tone became more youthful in appearance. In of about 0.001% to her neck once every four days. The addition, pigmentation become more even, for example, Subject also used Lancome makeup with Sunscreen and Oil freckles and other areas of increased pigmentation (e.g., of Olay moisturizer for sensitive skin. The subject reports melanin) decreased in color or disappeared. After 18 days of that since using the cytomodulin lotion, the wrinkles in her treatment, the difference in appearance between the treated neck were virtually eliminated. The subject reported that and untreated sides of the face was noticeable enough that there were no adverse effects. the Subject began to apply the topical composition to the entire face. No negative effects of application of the lotion 0.138 While preferred embodiments of the present inven were observed tion have been shown and described herein, it will be 0133. The same subject applied the CM-1 lotion to the obvious to those skilled in the art that such embodiments are back of one hand daily, while leaving the back of the other provided by way of example only. Numerous variations, hand untreated. The treatment continued for several weeks. changes, and Substitutions will now occur to those skilled in After this time, freckles and other pigmented areas on the the art without departing from the invention. It should be treated hand had essentially disappeared; there was no understood that various alternatives to the embodiments of change in the appearance of the untreated hand. In addition, the invention described herein may be employed in practic the skin looked plumper and thicker Smoother and shinier; ing the invention. It is intended that the following claims for example, veins appeared less prominent on the treated define the scope of the invention and that methods and skin. No negative effects of the application of the lotion were structures within the scope of these claims and their equiva observed. lents be covered thereby. US 2006/0293227 A1 Dec. 28, 2006 20

SEQUENCE LISTING

<160> NUMBER OF SEQ ID NOS: 44 <210> SEQ ID NO 1 &2 11s LENGTH 7 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 1

Ala Asn. Wall Ala Glu Asn Ala 1 5

<210> SEQ ID NO 2 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 2 Lieu. Ile Ala Glu Ala Lys 1 5

<210> SEQ ID NO 3 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 3

Lieu. Ile Ala Pro Glu Ala 1 5

<210> SEQ ID NO 4 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (2) <223> OTHER INFORMATION: alpha-amino isobutyric acid <400 SEQUENCE: 4 Leu Xaa Ala Glu Ala Lys 1 5

<210 SEQ ID NO 5 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (3) <223> OTHER INFORMATION: N-methyl alanine US 2006/0293227 A1 Dec. 28, 2006 21

-continued

<400 SEQUENCE: 5 Lieu. Ile Xaa Glu Ala Lys 1 5

<210> SEQ ID NO 6 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (2) <223> OTHER INFORMATION: alpha-amino butyric acid <400 SEQUENCE: 6 Leu Xaa Ala Glu Ala Lys 1 5

<210 SEQ ID NO 7 &2 11s LENGTH 7 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 7 Gly Gly Glin Ile Ala Asn. Ile 1 5

<210 SEQ ID NO 8 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 8 Glu Gly Ile Ala Gly Lys 1 5

<210 SEQ ID NO 9 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 9 Lieu. Ile Ala Asp Ala Lys 1 5

<210> SEQ ID NO 10 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 10 US 2006/0293227 A1 Dec. 28, 2006 22

-continued

Lieu. Ile Ala Asn Ala Lys 1 5

<210> SEQ ID NO 11 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 11

Lieu. Ile Ala Glu Ala Ala 1 5

<210> SEQ ID NO 12 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 12 Lieu. Ile Ala Glin Ala Lys 1 5

<210> SEQ ID NO 13 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 13 Lieu. Ile Ala Gly Gly Glu 1 5

<210> SEQ ID NO 14 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 14 Lieu. Ile Ala Gly Glu Gly 1 5

<210 SEQ ID NO 15 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 15 Ala Asn. Wall Ala Glu Lys 1 5

<210> SEQ ID NO 16 US 2006/0293227 A1 Dec. 28, 2006 23

-continued

&2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 16 Lieu. Ile Ala Lys Gly Lys 1 5

<210 SEQ ID NO 17 &2 11s LENGTH 15 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 17 Gly Thr Pro Gly Pro Glin Gly Ile Ala Gly Glin Arg Gly Val Val 1 5 10 15

<210> SEQ ID NO 18 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (2) <223> OTHER INFORMATION: alpha-amino isobutyric acid <400 SEQUENCE: 18 Ile Xaa Ala Glu Ala Lys 1 5

<210 SEQ ID NO 19 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MOD RES <222> LOCATION: (2) <223> OTHER INFORMATION: alpha-amino butyric acid <400 SEQUENCE: 19 Leu Xaa Ala Glu Ala Lys 1 5

<210> SEQ ID NO 20 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 20 Leu Pro Ala Glu Ala Lys 1 5 US 2006/0293227 A1 Dec. 28, 2006 24

-continued

<210> SEQ ID NO 21 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 21 Lieu. Ile Pro Glu Ala Lys 1 5

<210> SEQ ID NO 22 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (3) <223> OTHER INFORMATION: alpha-amino isobutyric acid <400 SEQUENCE: 22 Lieu. Ile Xaa Glu Ala Lys 1 5

<210> SEQ ID NO 23 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MOD RES <222> LOCATION: (4) <223> OTHER INFORMATION: alpha-amino isobutyric acid <400 SEQUENCE: 23

Lieu. Ile Ala Xala Glu Ala 1 5

<210> SEQ ID NO 24 &2 11s LENGTH 8 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (7) <223> OTHER INFORMATION: b-Alanine

<400 SEQUENCE: 24 Ile Trp Gly Lieu. Asp Gly Xaa Lys 1 5

<210> SEQ ID NO 25 &2 11s LENGTH 8 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide US 2006/0293227 A1 Dec. 28, 2006 25

-continued

&220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (7) <223> OTHER INFORMATION: b-Alanine

<400 SEQUENCE: 25 Trp Ile Ala Leu Glu Gly Xaa Lys 1 5

<210> SEQ ID NO 26 &2 11s LENGTH 9 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 26 Gly Pro Glin Gly Ile Ala Gly Glin Arg 1 5

<210 SEQ ID NO 27 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 27 Glin Gly Ile Ala Gly Glin 1 5

<210> SEQ ID NO 28 &2 11s LENGTH 7 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 28 Glin Gly Ile Ala Gly Glin Arg 1 5

<210 SEQ ID NO 29 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 29 Phe Gly Ile Ala Gly Phe 1 5

<210 SEQ ID NO 30 &2 11s LENGTH 5 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 30 US 2006/0293227 A1 Dec. 28, 2006 26

-continued

Gly Ile Ala Gly Glin 1 5

<210> SEQ ID NO 31 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 31 Glin Gly Ala Ile Ala Glin 1 5

<210> SEQ ID NO 32 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 32 Phe Gly Ile Ala Gly Phe 1 5

<210 SEQ ID NO 33 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 33 Cys Gly Ile Ala Gly Cys 1 5

<210> SEQ ID NO 34 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 34 Glu Gly Ile Ala Gly Lys 1 5

<210 SEQ ID NO 35 <211& LENGTH 4 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (1) <223> OTHER INFORMATION: variable amino acid

<400 SEQUENCE: 35

Xaa Ile Ala Ala US 2006/0293227 A1 Dec. 28, 2006 27

-continued

<210 SEQ ID NO 36 &2 11s LENGTH 3 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (3) <223> OTHER INFORMATION: variable amino acid

<400 SEQUENCE: 36

Ile Ala Xala 1

<210 SEQ ID NO 37 <211& LENGTH 4 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (1) <223> OTHER INFORMATION: variable amino acid &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (4) <223> OTHER INFORMATION: variable amino acid

<400 SEQUENCE: 37

Xaa Ile Ala Xala 1

<210 SEQ ID NO 38 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (3) <223> OTHER INFORMATION: N-methyl alanine <400 SEQUENCE: 38 Ile Ile Xaa Glu Ala Lys 1 5

<210 SEQ ID NO 39 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (3) <223> OTHER INFORMATION: D-alanine

<400 SEQUENCE: 39 Lieu. Ile Xaa Glu Ala Lys US 2006/0293227 A1 Dec. 28, 2006 28

-continued

<210> SEQ ID NO 40 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (4) <223> OTHER INFORMATION: D-glutamic acid

<400 SEQUENCE: 40 Lieu. Ile Ala Xala Ala Lys 1 5

<210> SEQ ID NO 41 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (5) <223> OTHER INFORMATION: D-glutamic acid &220s FEATURE <221 NAME/KEY: MOD RES <222> LOCATION: (5) <223> OTHER INFORMATION: variable amino acid

<400 SEQUENCE: 41

Lieu. Ile Ala Pro Xaa Ala 1 5

<210> SEQ ID NO 42 &2 11s LENGTH 6 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide &220s FEATURE <221 NAME/KEY: MOD RES <222> LOCATION: (4) <223> OTHER INFORMATION: trans-4-hydroxyproline &220s FEATURE <221 NAME/KEY: MODRES <222> LOCATION: (4) <223> OTHER INFORMATION: variable amino acid

<400 SEQUENCE: 42 Lieu. Ile Ala Xala Ala Lys 1 5

<210> SEQ ID NO 43 <211& LENGTH 4 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide US 2006/0293227 A1 Dec. 28, 2006 29

-continued

<400 SEQUENCE: 43

Lieu. Ile Ala Glu 1

<210> SEQ ID NO 44 <211& LENGTH 4 &212> TYPE PRT <213> ORGANISM: Artificial Sequence &220s FEATURE <223> OTHER INFORMATION: Description of Artificial Sequence: Synthetic peptide

<400 SEQUENCE: 44

Asn. Wall Ala Glu 1

1-27. (canceled) fatty ester emollient, and a compound selected from the 28. A method of cosmetic treatment of an individual group consisting of petrolatum and mineral oil. comprising topically administering to an individual Suffer 31. The method of claim 28 wherein the vehicle com ing from skin wrinkling a composition comprising a TGF-B prises water, mineral oil, isopropyl myristate, PEG-40 sor mimic of less than about 12 amino acids in length, in a bitan peroleate, glyceryllanolate, Sorbitol, propylene glycol, cosmetically acceptable vehicle at a concentration of about 0.0001% to about 0.01% by weight, wherein the TGF-f cetyl palmitate, magnesium Sulfate, aluminum Stearate, mimic comprises an amino acid sequence that is at least 80% lanolin alcohol, BHT methylchloroisothiazolinone, and identical to the amino acid sequence of SEQID NO: 1, and methylisothiazolinone wherein, if the sequence is not identical to SEQ ID NO: 1. 32. The method of claim 28 wherein the vehicle com it contains a single amino acid substitution. prises water, glycerin, distearyldimonium chloride, petrola 29. The method of claim 28 wherein the vehicle com tum, isopropyl palmitate, cetyl alcohol, dimethicone, prises emulsifying lanolin alcohols, waxes, and oils. Sodium chloride, methylparaben, and propylparaben. 30. The method of claim 28 wherein the vehicle com prises a quaternary ammonium compound, a fatty alcohol, a