Compositions containing cyclic peptides and methods of use

Abstract

The invention relates generally to cyclic peptides and their use in compositions, especially topical, cosmetic and/or personal care compositions, and compositions containing said cyclic peptides.

Claims

1. A method for reducing the visible signs of ageing of the skin in humans, wherein said visible signs of ageing of the skin are selected from the group consisting of stretch marks of the skin, wrinkles, skin lines, roughness of the skin, loss of skin elasticity, loss of skin firmness, loss of skin tightness, and slack tissue, comprising administering to a portion of the skin of said human showing said visible signs of ageing a composition for topical use and/or a cosmetic composition, said composition being a liposome composition comprising i) 0.001 to 1% by weight of cyclo-(Arg-Gly-Asp-DPhe-Acha) and/or a salt thereof, wherein at least 10% of the Cyclo-(Arg-Gly-Asp-DPhe-Acha) and/or the salt thereof is entrapped in liposomes, ii) 0.01 to 20% by weight of one or more lipids, iii) 60 to 99.99% by weight of one or more physiologically acceptable solvents, and optionally iv) 0.0001 to 20% by weight of one or more further ingredients, selected from the group consisting of ) further active compounds having a skin-care, hair-care and/or inflammation-inhibiting action, and ) further cosmetically acceptable excipients.

2. The method according to claim 1, wherein in the liposome composition, the lipids comprise one or more of the following: a) phospholipids, b) glycosphingolipids, c) lecithin, d) spingomyelin, e) dipalmitoyl lecithin, f) distearoylphosphatidylcholine, g) phosphatidylcholine, h) saturated phosphatidylcholine, i) unsaturated phosphatidylcholine, j) polystyrene, k) octyldodecanol, optionally in combination with Silica, l) octyldodecanol, optionally in combination with phospholipids, cholesterol and/or glycospingolipids, and/or one or more salts thereof.

3. The method according to claim 1, wherein in the liposome composition, the one or more physiologically acceptable solvents comprise: water, and/or an alcohol having 2 to 5 carbon atoms, which is ethanol, isopropanol, glycerin, 1,2-propylene glycol, 1,4-butanediol, pentylene glycol, sorbitol or a mixture thereof.

4. The method according to claim 1, wherein the liposome composition comprises i) 0.001 to 1% by weight of cyclo-(Arg-Gly-Asp-DPhe-Acha), and/or a salt thereof, ii) 0.01 to 20% by weight of one or more lipids, iii) 70 to 99% by weight of one or more physiologically acceptable solvents, and optionally iv) 0.001 to 10% by weight of one or more further ingredients, selected from the group consisting of ) further active compounds having a skin-care, hair-care and/or inflammation-inhibiting action, and ) further cosmetically acceptable excipients.

5. The method according to claim 1, wherein in the liposome composition, the further active compounds having a skin-care, hair-care and/or inflammation-inhibiting action comprises ectoin in an amount of 0.1 to 1% by weight.

6. The method according to claim 1, wherein in the liposome composition, the further active compounds having a skin-care, hair-care and/or inflammation-inhibiting action comprises ectoin in an amount of 1% by weight.

7. The method according to claim 1, wherein in the liposome composition, the further active compounds having a skin-care, hair-care and/or inflammation-inhibiting action comprises ectoin in an amount of up to 3% by weight.

8. The method according to claim 1, wherein in the liposome composition, at least 20% of the Cyclo-(Arg-Gly-Asp-DPhe-Acha) and/or the salts thereof is entrapped in liposomes.

9. The method according to claim 1, wherein in the liposome composition, at least 50% of the Cyclo-(Arg-Gly-Asp-DPhe-Acha) and/or the salts thereof is entrapped in liposomes.

10. The method according to claim 1, wherein in the liposome composition, at least 75% of the Cyclo-(Arg-Gly-Asp-DPhe-Acha) and/or the salts thereof is entrapped in liposomes.

11. The method according to claim 1, wherein in the liposome composition, at least 95% of the Cyclo-(Arg-Gly-Asp-DPhe-Acha) and/or the salts thereof is entrapped in liposomes.

12. The method according to claim 1, wherein in the liposome composition, the one or more lipids are selected from the group consisting of lecithin, spingomyelin, dipalmitoyl lecithin, distearoylphosphatidylcholine, phosphatidylcholine, saturated phosphatidylcholine and unsaturated phosphatidylcholine, or is a mixture thereof or is a salt thereof.

13. The method according to claim 1, wherein the liposome composition comprises 0.05 to 0.5% by weight of cyclo-(Arg-Gly-Asp-DPhe-Acha), and/or a salt thereof.

14. The method according to claim 1, wherein the liposome composition comprises 2 to 10% by weight of one or more lipids.

15. The method according to claim 1, wherein the liposome composition comprises 0.0001 to 20% by weight of one or more further ingredients, which are selected from the group consisting of ) further active compounds having a skin-care, hair-care and/or inflammation-inhibiting action, and ) further cosmetically acceptable excipients.

16. The method according to claim 1, wherein said visible signs of ageing of the skin is stretch marks of the skin.

17. The method according to claim 1, wherein said visible signs of ageing of the skin is wrinkles.

18. The method according to claim 1, wherein said visible signs of ageing of the skin is skin lines.

19. The method according to claim 1, wherein said visible signs of ageing of the skin is roughness of the skin.

20. The method according to claim 1, wherein said visible signs of ageing of the skin is loss of skin elasticity.

21. The method according to claim 1, wherein said visible signs of ageing of the skin is loss of skin firmness.

22. The method according to claim 1, wherein said visible signs of ageing of the skin is loss of skin tightness.

23. The method according to claim 1, wherein said visible signs of ageing of the skin is slack tissue.

Description

EXPERIMENTAL

(1) All temperatures stated above and below are in C. In the examples below, customary working up means: water is added if necessary, the mixture is neutralized and subjected to extraction with ether or dichloromethane, the phases are separated, the organic phase is dried over sodium sulfate, filtered and concentrated by evaporation and the residue is purified by chromatography on silica gel and/or crystallization. RT=retention time (minutes). Analysis was by HPLC on Lichrosorb RP select B (7 m)-2504 mm column, Eluent A: 0.3% TFA in water; Eluent B: 0.3% TFA in 2-propanol/water (8:2) gradient 1-99% B in 50 minutes at 1 ml/min flow rate and detection at 215 nm. M+=molecular peak in the mass spectrum obtained by the Fast Atom Bombardment (FAB) method.

Example 1

(2) A solution of 1.1 g of H-Arg(Mtr)-Gly-Asp(OBut)-D-Phe-hPro-ONa [obtainable, for example, from Fmoc-NMe-Arg(Mtr)-Gly-Asp(OBut)-D-Phe-hPro-O-Wang, -O-Wang being the radical of a 4-oxymethyl-phenoxymethyl-polystyrene resin used in the modified Merrifield techniques, by removal of the Fmoc group with piperidine/DMF and elimination of the resin with TFA/CH.sub.2Cl.sub.2(1:1)] in 15 ml of DMF is diluted with 85 ml of dichloromethane, and 50 mg of NaHCO.sub.3 are added. After cooling in a dry ice/acetone mixture, 40 l of diphenylphosphoryl azide are added. After standing at room temperature for 16 hours, the solution is concentrated. The concentrate is gel-filtered (Sephadex 010 column in isopropanol/water 8:2) and then purified by HPLC in the customary manner. Treatment with TFA/H.sub.2O (98:2) gives cyclo-(Arg-Gly-Asp-D-Phe-hPro); RT=18.5; FAB-MS (M+H): 587.

(3) The following are obtained analogously by cyclization of the corresponding linear peptides and removal of the protecting groups:

(4) cyclo-(Arg-Gly-Asp-DPhe-Nle);RT=25.3;FAB-MS(M+H):589;

(5) cyclo-(Arg-Gly-Asp-Phe-Ahds);RT=35.1;FAB-MS(M+H):730;

(6) cyclo-(Arg-Gly-Asp-DPhe-Ahds);RT=35.4;FAB-MS(M+H):730;

(7) cyclo-(Arg-Gly-Asp-Phe-DAhds);RT=35.7;FAB-MS(M+H):730;

(8) cyclo-(Arg-Gly-Asp-DPhe-Aos);

(9) cyclo-(Arg-Gly-Asp-DPhe-DAos);

(10) cyclo-(Arg-Gly-Asp-Phe-DAos);

(11) cyclo-(Arg-Gly-Asp-DPhe-Nhdg);RT=36.7;FAB-MS(M+H):758;

(12) cyclo-(Arg-Gly-Asp-Phe-Nhdg);RT=36.5;FAB-MS(M+H):758;

(13) cyclo-(Arg-Gly-Asp-DPhe-DNhdg);FAB-MS(M+H):758;

(14) cyclo-(Arg-Gly-Asp-Phe-DNhdg);FAB-MS(M+H):758;

(15) cyclo-(Arg-Gly-Asp-DPhg-Nhdg);

(16) cyclo-(Arg-Gly-Asp-Phg-Nhdg);

(17) cyclo-(Arg-Gly-Asp-DPhg-DNhdg);

(18) cyclo-(Arg-Gly-Asp-Phg-DNhdg);

(19) cyclo-(Arg-Gly-Asp-DPhe-Acha);RT=25.2;FAB-MS(M+H):601;

(20) cyclo-(Arg-Gly-Asp-Phe-Acha);FAB-MS(M+H):601;

(21) cyclo-(Arg-Gly-Asp-DPhe-DAcha);FAB-MS(M+H):601;

(22) cyclo-(Arg-Gly-Asp-Phe-DAcha);FAB-MS(M+H):601;

(23) cyclo-(Arg-Gly-Asp-DPhe-Aib);FAB-MS(M+H):575;

(24) cyclo-(Arg-Gly-Asp-Phe-Aib); RT=36.5;FAB-MS(M+H):575;

(25) cyclo-(Arg-Gly-Asp-DPhe-DAib);FAB-MS(M+H):575;

(26) cyclo-(Arg-Gly-Asp-Phe-DAib);FAB-MS(M+H):575;

(27) cyclo-(Arg-Gly-Asp-DPhe-Acpa);RT=17.1;FAB-MS(M+H):587;

(28) cyclo-(Arg-Gly-Asp-Phe-Acpa);FAB-MS(M+H):587;

(29) cyclo-(Arg-Gly-Asp-DPhe-DAcpa);FAB-MS(M+H):587;

(30) cyclo-(Arg-Gly-Asp-Phe-DAcpa);FAB-MS(M+H):587;

(31) cyclo-(Arg-Gly-Asp-DPhe-Tle);RT=19.1;FAB-MS(M+H):589;

(32) cyclo-(Arg-Gly-Asp-Phe-Tle);FAB-MS(M+H):589;

(33) cyclo-(Arg-Gly-Asp-DPhe-DTIe);FAB-MS(M+H):589;

(34) cyclo-(Arg-Gly-Asp-Phe-DTIe);FAB-MS(M+H):589;

(35) cyclo-(Arg-Gly-Asp-Dphe(4-Cl)-Tle);RT=23.2;FAB-MS(M+H):623;

(36) cyclo-(Arg-Gly-Asp-Phe(4-Cl)-Tle);FAB-MS(M+H):623;

(37) cyclo-(Arg-Gly-Asp-DPhe(4-Cl)-DTIe);FAB-MS(M+H):623;

(38) cyclo-(Arg-Gly-Asp-Phe(4-Cl)-DTIe);FAB-MS(M+H):623;

(39) cyclo-(Arg-Gly-Asp-Dphe(4-F)-Tle);RT=20.2;FAB-MS(M+H):607;

(40) cyclo-(Arg-Gly-Asp-Phe(4-F)-Tle);FAB-MS(M+H):607;

(41) cyclo-(Arg-Gly-Asp-DPhe(4-F)-DTIe);FAB-MS(M+H):607;

(42) cyclo-(Arg-Gly-Asp-Phe(4-F)-DTIe);FAB-MS(M+H):607.

Example 2

(43) A solution of 0.28 g of cyclo-(Arg(Mtr)-Gly-Asp-DPhe-DhPro)[obtainable by cyclization according to Ex. 1] in 8.4 ml of TFA, 1.7 ml of dichloromethane and 0.9 ml of thiophenol is allowed to stand at room temperature for 4 hours, then concentrated, and the residue is diluted with water and then freeze-dried. Gel filtration on Sephadex G 10 (acetic acid/water 1:1) and subsequent purification by preparative HPLC under the conditions indicated give cyclo-(Arg-Gly-Asp-DPhe-DhPro); FAB-MS (M+H): 587.

(44) The following are obtained analogously:

(45) from cyclo-(Arg(Mtr)-Gly-Asp-Phe-DhPro):

(46) cyclo-(Arg-Gly-Asp-Phe-DhPro); FAB-MS (M+H): 587;
from cyclo-(Arg(Mtr)-Gly-Asp(OBut)-DPhg-Tle): cyclo-(D-Arg-NMeGly-Asp-DPhg-Tle);
from cyclo-(Arg(Mtr)-Gly-Asp(OEt)-DPhg-hPro): cyclo-(Arg-Gly-Asp-DPhg-h Pro);
from cyclo-(Arg(Mtr)-Gly-Asp-Phg-DAhds): cyclo-(Arg-Gly-Asp-Phg-DAhds);
from cyclo-(Arg(Mtr)-Gly-Asp-DPhg-Acpa): cyclo-(Arg-Gly-Asp-DPhg-Acpa);
from cyclo-(Arg(Mtr)-Gly-Asp-DPhg-Aos): cyclo-(Arg-Gly-Asp-DPhg-Aos).

Example 3

(47) 80 mg of cyclo-(Arg-Gly-Asp-DPhe-hPro) [obtainable according to Ex. 1] are dissolved in 0.01 m HCl five to six times and freeze-dried after each dissolving operation. Subsequent purification by HPLC gives cyclo-(Arg-Gly-Asp-DPhe-hPro)HCl.

(48) The following are obtained analogously:

(49) from cyclo-(Arg-Gly-Asp-DPhe-Nle):

(50) cyclo-(NMeArg-Gly-Asp-DPhe-Nle)HCl;
from cyclo-(Arg-Gly-Asp-DPhe-Ahds): cyclo-(Arg-Gly-Asp-DPhe-Ahds)HCl;
from cyclo-(Arg-Gly-Asp-DPhe-Ahds): cyclo-(Arg-Gly-Asp-DPhe-Ahds)HCl.

Example 4

(51) To prepare affinity phases, 0.9 g of N-maleimido-(CH.sub.2).sub.5CONH(CH.sub.2).sub.3 polymer [obtainable by condensation of N-maleimido-(CH.sub.2).sub.5COOH with H.sub.2N(CH.sub.2).sub.3 polymer] is suspended in 10 ml of 0.1 M sodium phosphate buffer at a pH of 7, and one equivalent cyclo-(Arg-Gly-Asp-DPhe(4-NCO(CH.sub.2).sub.2SH)-hPro) [obtainable by cyclization of H-Dphe(4-NH-BOC)-hPro-Arg(Mtr)-Gly-Asp-OH, removal of the protecting groups and acylation with, for example, ClCO(CH.sub.2).sub.2SH] is added at 4. The reaction mixture is stirred for 4 hours with simultaneous warming to room temperature, and the solid residue is filtered off and washed twice with 10 ml each of buffer solution (pH 7) and then three times with 10 ml each time of water. Cyclo-(Arg-Gly-Asp-DPhe(4-NCO(CH.sub.2).sub.2S-3-(N-maleimido-(CH.sub.2).sub.5CONH(CH.sub.2).sub.3-polymer)-hPro)) is obtained.

Example 5

(52) Analogously to Example 4, condensation of polymer-O(CH.sub.2).sub.3NH.sub.2 [commercially available] and cyclo-(Arg-Gly-Asp-DPhe(4-NCO(CH.sub.2).sub.4COOH)=hPro) [obtainable by condensing adipic acid with cyclo-(Arg(Mtr)-Gly-Asp-DPhe-(4-NH-BOC)-hPro) under the conditions stated in Ex. 4] gives the following polymeric phase: cyclo-(Arg-Gly-Asp-DPhe(4-NCO(CH.sub.2).sub.4CONH(CH.sub.2).sub.3O-polymer)-hPro.

(53) In the examples below, customary working up means: water is added if necessary, the mixture is neutralized and subjected to extraction with ether or dichloroethane, the phases are separated, the organic phase is dried over sodium sulfate, filtered and concentrated by evaporation and the residue is purified by chromatography on silica gel and/or crystallization. RT=retention time (minutes). Analysis was by HPLC on Lichrosorb RP select B (7 m)-2504 mm column, Eluent A: 0.3% TFA in water; Eluent B: 0.3% TFA in 2-propanol/water (8:2) gradient 1-99% B in 50 minutes at 1 ml/min flow rate and detection at 215 nm. M+=molecular peak in the mass spectrum obtained by the Fast Atom Bombardment (FAB) method.

Example 6

(54) A solution of 0.6 g of H-NMe-Arg(Mtr)-Gly-Asp(OBut)-D-Phe-Val-ONa [obtainable for example from Fmoc-NMe-Arg(Mtr)-Gly-Asp(OBut)-D-Phe-Val-O-Wang, -O-Wang being the radical of a 4-hydroxymethyl-phenoxymethyl-polystyrene resin used in the modified Merrifield techniques, by removal of the Fmoc group with piperidine/DMF and elimination of the resin with TFA/CH.sub.2Cl.sub.2 (1:1)] in 15 ml of DMF is diluted with 85 ml of dichloromethane, and 50 mg of NaHCO.sub.3 are added. After cooling in a dry ice/acetone mixture, 40 III of diphenylphosphoryl azide are added. After standing at room temperature for 16 hours, the solution is concentrated. The concentrate is gel-filtered (Sephadex 010 column in isopropanol/water 8:2) and then purified by HPLC in the customary manner. Treatment with TFA/H.sub.2 0 (98:2) gives cyclo-(NMe-Arg-Gly-Asp-D-Phe-Val); RT=18.1; FAB-MS (M+H): 589.

(55) The following are obtained analogously by cyclization of the corresponding linear peptides and elimination of the protecting groups:

(56) cyclo-(Arg-NMeGly-Asp-DPhe-Val); RT=17.9; FAB-MS (M+H): 589;

(57) cyclo-(Arg-Gly-NMeAsp-DPhe-Val); RT=18.3; FAB-MS (M+H): 589;

(58) cyclo-(Arg-Gly-NMeAsp-DPhe-Val)TFA; RT=15.4; FAB-MS(M+H): 589;

(59) cyclo-(Arg-Gly-Asp-NMeDPhe-Val); RT=18.9; FAB-MS (M+H): 589;

(60) cyclo-(Arg-Gly-Asp-DPhe-NMeVal); RT=19.5; FAB-MS (M+H): 589;

(61) cyclo-(Arg-Gly-Asp-DPhe-NMeLys); RT=11.1; FAB-MS(M+H): 618;

(62) cyclo-(Arg-Gly-Asp-DPhe-NMeLys(benzyloxycarbonyl)TFA=23.4; FAB-MS (M+H): 752;

(63) cyclo-(NEtArg-Gly-Asp-DPhe-Val); FAB-MS (M+H): 603;

(64) cyclo-(Arg-NEtGly-Asp-DPhe-Val); FAB-MS (M+H): 603;

(65) cyclo-(Arg-Gly-NEtAsp-DPhe-Val); FAB-MS (M+H): 603;

(66) cyclo-(Arg-Gly-Asp-NEtDPhe-Val); FAB-MS (M+H): 603;

(67) cyclo-(Arg-Gly-Asp-DPhe-NEtVal); FAB-MS (M+H): 603;

(68) cyclo-(Arg-Gly-Asp-DPhe(4-I)-NMeVal); RT=23.5; FAB-MS(M+H): 715;

(69) cyclo-(NPrArg-Gly-Asp-DPhe-Val); FAB-MS (M+H): 617;

(70) cyclo-(Arg-NPrGly-Asp-DPhe-Val); FAB-MS (M+H): 617;

(71) cyclo-(Arg-Gly-NPrAsp-DPhe-Val); FAB-MS (M+H): 617;

(72) cyclo-(Arg-Gly-Asp-NPrDPhe-Val); FAB-MS (M+H): 617;

(73) cyclo-(Arg-Gly-Asp-DPhe-NPrVal); FAB-MS (M+H): 617;

(74) cyclo-(NBzlArg-Gly-Asp-DPhe-Val); FAB-MS (M+H): 665;

(75) cyclo-(Arg-NBzlGly-Asp-DPhe-Val); FAB-MS (M+H): 665;

(76) cyclo-(Arg-Gly-NBzlAsp-DPhe-Val); FAB-MS (M+H): 665;

(77) cyclo-(Arg-Gly-Asp-NBzlDPhe-Val); FAB-MS (M+H): 665;

(78) cyclo-(Arg-Gly-Asp-DPhe-NBzlVal); FAB-MS (M+H): 665;

(79) cyclo-(Arg-Gly-Asp-Phe-DNMeVal)TFA; RT=18.2; FAB-MS(M+H): 589;

(80) cyclo-(NMeArg-Gly-Asp-DPhe-Leu); FAB-MS (M+H): 603;

(81) cyclo-(Arg-NMeGIy-Asp-DPhe-Leu); FAB-MS (M+H): 603;

(82) cyclo-(Arg-Gly-NMeAsp-DPhe-Leu); FAB-MS (M+H): 603;

(83) cyclo-(Arg-Gly-Asp-NMeDPhe-Leu); FAB-MS (M+H): 603;

(84) cyclo-(Arg-Gly-Asp-DPhe-NMeLeu); FAB-MS (M+H): 603;

(85) cyclo-(NEtArg-Gly-Asp-DPhe-Leu); FAB-MS (M+H): 617;

(86) cyclo-(Arg-NEtGIy-Asp-DPhe-Leu); FAB-MS (M+H): 617;

(87) cyclo-(Arg-Gly-NEtAsp-DPhe-Leu); FAB-MS (M+H): 617;

(88) cyclo-(Arg-Gly-Asp-NEtDPhe-Leu); FAB-MS (M+H): 617;

(89) cyclo-(Arg-Gly-Asp-DPhe-NEtLeu); FAB-MS (M+H): 617;

(90) cyclo-(NPrArg-Gly-Asp-DPhe-Leu); FAB-MS (M+H): 631;

(91) cyclo-(Arg-NPrGIy-Asp-DPhe-Leu); FAB-MS (M+H): 631;

(92) cyclo-(Arg-Gly-NPrAsp-DPhe-Leu); FAB-MS (M+H): 631;

(93) cyclo-(Arg-Gly-Asp-NPrDPhe-Leu); FAB-MS (M+H): 631;

(94) cyclo-(Arg-Gly-Asp-DPhe-NPrLeu); FAB-MS (M+H): 631;

(95) cyclo-(NBzlArg-Gly-Asp-DPhe-Leu); FAB-MS (M+H): 679;

(96) cyclo-(Arg-NBzlGly-Asp-DPhe-Leu); FAB-MS (M+H): 679;

(97) cyclo-(Arg-Gly-NBzlAsp-DPhe-Leu); FAB-MS (M+H): 679;

(98) cyclo-(Arg-Gly-Asp-NBzlDPhe-Leu); FAB-MS (M+H): 679;

(99) cyclo-(Arg-Gly-Asp-DPhe-NBzlLeu); FAB-MS (M+H): 679;

(100) cyclo-(NMeArg-Gly-Asp-DPhe-Ala);

(101) cyclo-(Arg-NMeGly-Asp-DPhe-Ala);

(102) cyclo-(Arg-Gly-NMeAsp-DPhe-Ala);

(103) cyclo-(Arg-Gly-Asp-NMeDPhe-Ala);

(104) cyclo-(Arg-Gly-Asp-DPhe-NMeAla); RT=16.2; FAB-MS (M+H): 561;

(105) cyclo-(NEtArg-Gly-Asp-DPhe-Ala);

(106) cyclo-(Arg-NEtGly-Asp-DPhe-Ala);

(107) cyclo-(Arg-Gly-NEtAsp-DPhe-Ala);

(108) cyclo-(Arg-Gly-Asp-N EtD Phe-Ala);

(109) cyclo-(Arg-Gly-Asp-DPhe-NEtAla);

(110) cyclo-(N PrArg-Gly-Asp-DPhe-Ala);

(111) cyclo-(Arg-N PrGly-Asp-DPhe-Ala);

(112) cyclo-(Arg-Gly-NPrAsp-DPhe-Ala);

(113) cyclo-(Arg-Gly-Asp-NPrDPhe-Ala);

(114) cyclo-(Arg-Gly-Asp-DPhe-N PrAla);

(115) cyclo-(NBzlArg-Gly-Asp-DPhe-Ala);

(116) cyclo-(Arg-NBzlGly-Asp-DPhe-Ala);

(117) cyclo-(Arg-Gly-NBzlAsp-DPhe-Ala);

(118) cyclo-(Arg-Gly-Asp-NBzlDPhe-Ala);

(119) cyclo-(Arg-Gly-Asp-DPhe-NBzlAla);

(120) cyclo-(NMeArg-Gly-Asp-DPhe-Gly);

(121) cyclo-(Arg-NMeGly-Asp-DPhe-Gly);

(122) cyclo-(Arg-Gly-NMeAsp-DPhe-Gly);

(123) cyclo-(Arg-Gly-Asp-NMeDPhe-Gly);

(124) cyclo-(Arg-Gly-Asp-DPhe-NMeGly); RT=14.3; FAB-MS(M+H): 547;

(125) Cyclo-(DArg-Gly-Asp-DPhe-NMeVal)TFA; RT=18.7; FAB-MS(M+H): 589;

(126) cyclo-(NEtArg-Gly-Asp-DPhe-Gly);

(127) cyclo-(Arg-NEtGly-Asp-DPhe-Gly);

(128) cyclo-(Arg-Gly-NEtAsp-DPhe-Gly);

(129) cyclo-(Arg-Gly-Asp-NEtDPhe-Gly);

(130) cyclo-(Arg-Gly-Asp-DPhe-NEtGly);

(131) cyclo-(NPrArg-Gly-Asp-DPhe-Gly);

(132) cyclo-(Arg-NPrGly-Asp-DPhe-Gly);

(133) cyclo-(Arg-Gly-NPrAsp-DPhe-Gly);

(134) cyclo-(Arg-Gly-Asp-NPrDPhe-Gly);

(135) cyclo-(Arg-Gly-Asp-DPhe-NPrGly);

(136) cyclo-(NBzlArg-Gly-Asp-DPhe-Gly);

(137) cyclo-(Arg-NBzlGly-Asp-DPhe-Gly);

(138) cyclo-(Arg-Gly-NBzlAsp-DPhe-Gly);

(139) cyclo-(Arg-Gly-Asp-NBzlDPhe-Gly);

(140) cyclo-(Arg-Gly-Asp-DPhe-NBzlGly);

(141) cyclo-(NMeArg-Gly-Asp-Phg-Val) (SEQ ID NO: 1);

(142) cyclo-(Arg-NMeGly-Asp-Phg-Val) (SEQ ID NO: 2);

(143) cyclo-(Arg-Gly-NMeAsp-Phg-Val) (SEQ ID NO: 3);

(144) cyclo-(Arg-Gly-Asp-NMePhg-Val) (SEQ ID NO: 4);

(145) cyclo-(Arg-Gly-Asp-Phg-NMeVal) (SEQ ID NO: 5);

(146) cyclo-(NEtArg-Gly-Asp-Phg-Val) (SEQ ID NO: 6);

(147) cyclo-(Arg-NEtGly-Asp-Phg-Val) (SEQ ID NO: 7);

(148) cyclo-(Arg-Gly-NEtAsp-Phg-Val) (SEQ ID NO: 8);

(149) cyclo-(Arg-Gly-Asp-NEtPhg-Val) (SEQ ID NO: 9);

(150) cyclo-(Arg-Gly-Asp-Phg-NEtVal) (SEQ ID NO: 10);

(151) cyclo-(NPrArg-Gly-Asp-Phg-Val) (SEQ ID NO: 11);

(152) cyclo-(Arg-NPrGly-Asp-Phg-Val) (SEQ ID NO: 12);

(153) cyclo-(Arg-Gly-NPrAsp-Phg-Val) (SEQ ID NO: 13);

(154) cyclo-(Arg-Gly-Asp-NPrPhg-Val) (SEQ ID NO: 14);

(155) cyclo-(Arg-Gly-Asp-Phg-NPrVal) (SEQ ID NO: 15);

(156) cyclo-(NBzlArg-Gly-Asp-Phg-Val) (SEQ ID NO: 16);

(157) cyclo-(Arg-NBzlGly-Asp-Phg-Val) (SEQ ID NO: 17);

(158) cyclo-(Arg-Gly-NBzlAsp-Phg-Val) (SEQ ID NO: 18);

(159) cyclo-(Arg-Gly-Asp-NBzlPhg-Val) (SEQ ID NO: 19);

(160) cyclo-(Arg-Gly-Asp-Phg-NBzlVal) (SEQ ID NO: 20);

(161) cyclo-(NMeArg-Gly-Asp-Trp-Val) (SEQ ID NO: 21);

(162) cyclo-(Arg-NMeGly-Asp-Trp-Val) (SEQ ID NO: 22);

(163) cyclo-(Arg-Gly-NMeAsp-Trp-Val) (SEQ ID NO: 23);

(164) cyclo-(Arg-Gly-Asp-NMeTrp-Val) (SEQ ID NO: 24);

(165) cyclo-(Arg-Gly-Asp-Trp-NMeVal) (SEQ ID NO: 25);

(166) cyclo-(NEtArg-Gly-Asp-Trp-Val) (SEQ ID NO: 26);

(167) cyclo-(Arg-NEtGly-Asp-Trp-Val) (SEQ ID NO: 27);

(168) cyclo-(Arg-Gly-NEtAsp-Trp-Val) (SEQ ID NO: 28);

(169) cyclo-(Arg-Gly-Asp-NEtTrp-Val) (SEQ ID NO: 29);

(170) cyclo-(Arg-Gly-Asp-Trp-NEtVal) (SEQ ID NO: 30);

(171) cyclo-(NPrArg-Gly-Asp-Trp-Val) (SEQ ID NO: 31);

(172) cyclo-(Arg-NPrGly-Asp-Trp-Val) (SEQ ID NO: 32);

(173) cyclo-(Arg-Gly-NPrAsp-Trp-Val) (SEQ ID NO: 33);

(174) cyclo-(Arg-Gly-Asp-NPrTrp-Val) (SEQ ID NO: 34);

(175) cyclo-(Arg-Gly-Asp-Trp-NPrVal) (SEQ ID NO: 35);

(176) cyclo-(NBzlArg-Gly-Asp-Trp-Val) (SEQ ID NO: 36);

(177) cyclo-(Arg-NBzGlyy-Asp-Trp-Val) (SEQ ID NO: 37);

(178) cyclo-(Arg-Gly-NBzlAsp-Trp-Val) (SEQ ID NO: 38);

(179) cyclo-(Arg-Gly-Asp-NBzlTrp-Val) (SEQ ID NO: 39);

(180) cyclo-(Arg-Gly-Asp-Trp-NBzlVal) (SEQ ID NO: 40).

Example 7

(181) A solution of 0.28 g of cyclo-(Arg(Mtr)-Gly-Asp-NMePhe-Dval) [obtainable by cyclization according to Ex. 1] in 8.4 ml of TFA, 1.7 ml of dichloromethane and 0.9 ml of thiophenol is allowed to stand at room temperature for 4 hours, then concentrated, and the residue is diluted with water and then freeze-dried. Gel filtration on Sephadex G 10 (acetic acid/water 1:1) and subsequent purification by preparative HPLC under the conditions indicated give cyclo-(Arg-Gly-Asp-NMePhe-DVal); FAB-MS (M+H): 589.

(182) The following are obtained analogously:

(183) from cyclo-(Arg(Mtr)-Gly-NMeAsp-DPhe-Ile): cyclo-(Arg-Gly-NMeAsp-DPhe-Ile); FAB-MS (M+H): 603;

(184) from cyclo-(D-Arg(Mtr)-NMeGly-Asp(OBut)-DPhe-Nle): cyclo-(D-Arg-NMeGly-Asp-DPhe-Nle);

(185) from cyclo-(NMeArg(Mtr)-Gly-D-Asp(OEt)-DPhe-Ile): cyclo-(NMeArg-Gly-D-Asp-DPhe-Ile);

(186) from cyclo-(NMeArg(Mtr)-Gly-Asp-Phe-DIle): cyclo-(NMeArg-Gly-Asp-Phe-DIle);

(187) from cyclo-(Arg(Mtr)-Gly-NMeAsp-Phe-DLeu): cyclo-(Arg-Gly-NMeAsp-Phe-DLeu);

(188) from cyclo-(Arg(Mtr)-NMeGly-Asp-Phe-DSer): cyclo-(Arg-NMeGIy-Asp-Phe-DSer);

(189) from cyclo-(Arg(Mtr)-NMeGly-Asp-DNal-Leu): cyclo-(Arg-NMeGly-Asp-DNal-Leu);

(190) from cyclo-(NMeArg(Mtr)-Gly-Asp-Nal-DIle): cyclo-(NMeArg-Gly-Asp-Nal-DIle);

(191) from cyclo-(Arg(Mtr)-Gly-Asp-NMePhg-DVal): cyclo-(Arg-Gly-Asp-NMePhg-DVal);

(192) from cyclo-(Arg(Mtr)-Gly-NMeAsp-Trp-DVal): cyclo-(Arg-Gly-NMeAsp-Trp-DVal).

Example 8

(193) 80 mg of cyclo-(Arg-Gly-Asp-DPhe-NMeVal) are dissolved in 0.01 m HCl five to six times and freeze-dried after each dissolving operation. Subsequent purification by HPLC gives cyclo-(Arg-Gly-Asp-DPhe-NMeVal)HCl; FAB-MS (M+H): 589.

(194) The following are obtained analogously:

(195) from cyclo-(NMeArg-Gly-Asp-DPhe-Val): cyclo-(NMeArg-Gly-Asp-DPhe-Val)HCl;

(196) from cyclo-(Arg-NMeGly-Asp-DPhe-Val): cyclo-(Arg-NMeGly-Asp-DPhe-Val)HCl; FAB-MS (M+H): 589;

(197) from cyclo-(Arg-Gly-NMeAsp-DPhe-Val): cyclo-(Arg-Gly-NMeAsp-DPhe-Val)HCl;

(198) from cyclo-(Arg-Gly-Asp-NMeDPhe-Val): cyclo-(Arg-Gly-Asp-NMeDPhe-Val)HCl;

(199) from cyclo-(Arg-Gly-Asp-Phe-DNMeVal): cyclo-(Arg-Gly-Asp-Phe-DNMeVal)HCl; RT=18.2; FAB-MS(M+H): 589.

(200) Analogously the following is obtained by the treatment with acetic acid (AcOH):

(201) from cyclo-(Arg-Gly-NMeAsp-DPhe-Val): cyclo-(Arg-Gly-NMeAsp-DPhe-Val)AcOH; RT=15.4; FAB-MS(M+H): 589.

(202) Analogously the following is obtained by the treatment with methane sulfonic acid (MeSO.sub.3H):

(203) from cyclo-(Arg-Gly-Asp-DPhe-NMeVal): cyclo-(Arg-Gly-Asp-DPhe-NMeVal)MeSO.sub.3H; RT=17.8; FAB-MS(M+H): 589;

Example 9

(204) To prepare affinity phases, 0.9 g of N-maleimido-(CH.sub.2).sub.5CONH(CH.sub.2).sub.3 polymer [obtainable by condensation of N-maleimido-(CH.sub.2).sub.5COOH with H.sub.2N(CH.sub.2).sub.3 polymer] is suspended in 10 ml of 0.1 M sodium phosphate buffer at a pH of 7, and one equivalent cyclo-(Arg-Gly-Asp-DPhe-NMeLys(CO(CH.sub.2).sub.2SH) is added at 4. The reaction mixture is stirred for 4 hours with simultaneous warming to room temperature, and the solid residue is filtered off and washed twice with 10 ml each of buffer solution (pH 7) and then three times with 10 ml each time of water. Cyclo-(Arg-Gly-Asp-DPhe-NMeLys(CO(CH.sub.2).sub.2S-3-(N-maleimido-(CH.sub.2).sub.5CONH(CH.sub.2).sub.3 polymer)) is obtained.

Example 10

(205) Analogously to Example 9, condensation of polymer-O(CH.sub.2).sub.3NH.sub.2 [commercially available] and cyclo-(Arg-Gly-Asp-NMe-DPhe-Lys(CO(CH.sub.2).sub.4COOH) [obtainable by condensing adipic acid with cyclo-(Arg-Gly-Asp-NMe-DPhe-Lys) under the stated conditions] gives the following polymeric phase: cyclo-(Arg-Gly-Asp-NMe-DPhe-Lys-(C0-(CH.sub.2).sub.4CONH(CH.sub.2).sub.3O-polymer)

(206) The following is obtained analogously by condensation of:

(207) cyclo-(NMe-Arg-Gly-Asp-DPhe-Lys-(CO(CH.sub.2).sub.5NH.sub.2)) with HOOCCH.sub.2O-polymer:

(208) cyclo-(NMe-Arg-Gly-Asp-DPhe-Lys-(C0-(CH.sub.2).sub.5NHCOCH.sub.2O-polymer)).

Example 11

(209) Biological Activities

(210) i) Isolated Integrin-Ligand Binding Assay

(211) The production of recombinant human integrin is known in the literature. The inhibitory activity of the substances given above are tested in a ligand inhibition assay, using immobilized integrin as the target, and biotinylated human serum vitronectin for a.sub.V.sub.3 as ligand. In brief, 96-well ELISA plates are coated by adsorption from neutral aqueous buffers of 1 microg/ml integrin. After blocking residual sites on the plate with BSA, biotinylated ligands (1 microg/ml) are added in the presence or absence of serial dilutions of inhibitor, and after incubation and washing, bound biotin is detected with peroxidase-coupled anti-biotin antibody and TMB substrate. IC.sub.50, the concentration of inhibitor needed to inhibit ligand binding in the absence of inhibitor by 50%, is established by curve fitting, and the values presented are usually the mean of three or more such independent determinations.

(212) Cyclo-(Arg-Gly-Asp-DPhe-(NMe)Ala)=cyclo(RGDf(NMe)A):

(213) IC.sub.50 on integrin .sub.V.sub.3 is 24 nM

(214) Cyclo-(Arg-Gly-Asp-DPhe-(NMe)Val)=cyclo(RGDf(NMe)V): IC.sub.50 on integrin .sub.V.sub.3 is 3 nM

(215) ii) Receptor Inhibition Assay

(216) Purified human integrin .sub.V.sub.3 from term placenta is adsorbed to microtitre wells and challenged with biotinylated complementary ligandsvitronectin (VN) for .sub.V.sub.3 in the presence of increasing amounts of test compounds.

(217) Method: 1 g ml.sup.1 biotin-ligand is incubated with 1 g ml.sup.1 coated receptor in the presence of serially diluted peptides. After 3 h at 30 C. bound ligand was measures by anti-biotinalkaline phosphatase detection.

(218) Literature: Charo, I. F., Nannizzi, L., Smith, J. W. and Cheresh, D. A., J. Cell. Biol. 111, 2795-2800 (1990).

(219) IC.sub.50 values for binding of biotinylated ligands to human placental .sub.V.sub.3

(220) TABLE-US-00001 IC.sub.50 [nM] Sequence VN: .sub.v.sub.3 Cyclo-(Arg-Gly-Asp-DPhe-Aib) 20 Cyclo-(Arg-Gly-Asp-DPhe-Acpa) 9 Cyclo-(Arg-Gly-Asp-DPhe-hPro) 170 Cyclo-(Arg-Gly-Asp-Phe-Nhdg) 8 Cyclo-(Arg-Gly-Asp-DPhe-Acha) 2 Cyclo-(Arg-Gly-Asp-DPhe-Tle) 6 Cyclo-(Arg-Gly-Asp-DPhe(4-Cl)-Tle) 1.5 Cyclo-(Arg-Gly-Asp-DPhe(4-F)-Tle) 3 Cyclo(Arg-Gly-Asp-Phe-Gly) 400
iii) Pentapeptides as Inhibitors of .sub.V.sub.3 (Immobilized)

(221) Preparation and Characterization of Integrin .sub.V.sub.3: Integrin .sub.V.sub.3 is purified from human placental extracts by affinity chromatography on GRGDSPK peptide. Extraction and chromatography follows previously published protocols except that Triton X-100 is replaced by 25 mM octyl-P-D-glucopyranoside. Bound integrin is eluted with 10 mM EDTA into 1.5-ml vials containing 25 pl of 1 M MgCl, and concentrated in Centricon 100 microconcentrators. They are stored at 4 C. in neutral buffer containing 25 mM octyl glucoside, and 0.05% sodium azide. Protein concentrations were determined by a micro BCA assay (Pierce). The purified integrin is characterized by SDS-gel electrophoresis followed by protein staining and by immunoblotting, which demonstrates the purity and identity of the two subunits.

(222) Solid-phase inhibition assays are established for the inhibition of binding of the vitronectin receptor a.sub.V.sub.3 to their corresponding protein ligands by the cyclic peptides. Inhibitory activities (IC.sub.50) of cyclic RGD containing peptides for the binding of vitronectin (VN). Integrins are used as immobilized ligands.

(223) TABLE-US-00002 IC.sub.50 [M] Sequence VN: .sub.v.sub.3 Cyclo-(DArg-Gly-Asp-Phe-Val) (=cyclo(rGDFV)) 72 Cyclo-(Arg-DAla-Asp-Phe-Val) (=cyclo(RaDFV)) >240 Cyclo-(Arg-Gly-DAsp-Phe-Val) (=cyclo(RGdFV)) 152 Cyclo-(Arg-Gly-Asp-DPhe-Val) (=cyclo(RGDfV)) 0.049 Cyclo-(Arg-Ala-Asp-DPhe-Val) (=cyclo(RADfV)) 4 Cyclo-(Arg-Gly-Asp-Phe-DVal) (=cyclo(RGDFv)) 11
iv) Integrin Inhibitor Characterisation

(224) The biological activities of the following RGD peptides Cyclo-(Arg-Gly-Asp-DPhe-(NMe)Val), Cyclo-(Arg-Gly-Asp-DPhe-Val), Gly-Arg-Gly-Asp-Ser-Pro-Lys-OH and Cyclo-(Arg-Gly-Asp-DPhe-Acha) were compared in an isolated integrin binding assay as detailed elsewhere. Briefly, in receptor studies, recombinant human .sub.3 integrin is adsorbed onto 96-well plates, and biotinylated human native ligand (plasma vitronectin) is added in the presence of serially diluted RGD peptides. After 3 hours at 37 C., bound ligand is detected using alkaline-phosphatase labelled anti-biotin MAbs (Sigma, St. Louis, Mo.).

(225) Gly-Arg-Gly-Asp-Ser-Pro-Lys-OH: IC.sub.50=120 nM

(226) Cyclo-(Arg-Gly-Asp-DPhe-Val): IC.sub.50=1.5 nM

(227) Cyclo-(Arg-Gly-Asp-DPhe-(NMe)Val): IC.sub.50=5.1 nM

(228) Cyclo-(Arg-Gly-Asp-DPhe-Acha): IC.sub.50=2.1 nM

Example 12: cDNA-Microarray Studies

(229) cDNA-Microarray Studies

(230) The following is a study to determine the gene expression profiles of Cyclo-(Arg-Gly-Asp-DPhe-Acha) (=CPPC 1) treated human skin.

(231) For the analysis of deregulated gene expression induced by substance CPPC 1, treated and control samples from a human skin model are analyzed as follows: A full thickness skin model (human primary epidermal keratinocytes and human fibroblasts), such as the Phenion FTSM, commercially available from Phenion GmbH & Co. KG, Dsseldorf, Germany, is used. Cells are not pooled or genetically modified. Gene expression analysis is carried out with PIQOR-skin cDNA microarrays, which contains 1312 genes that are involved in target pathways related to stress, inflammation, pigmentation and depigmentation, moisturization, anti-ageing and hair follicle development in humans. For instance, cell cycle, apoptosis, DNA repair, oxidative metabolism, angiogenesis, cell adhesion cell-matrix interactions and signaling. Genes are represented with four replicate spots. Treatment with CPPC 1 is performed in 0.5 M concentration (4 days incubation, n=3 per condition). Buffer-treated skin equivalents serve as controls. Selection of genes is based on statistical methods (p<0.05, at least 1.5-fold deregulation). 345 deregulated genes are selected for 0.5 M.
Profiling of Gene ExpressionInput Genes

(232) A ratio (log 2) value of zero indicates no regulation, whereas positive (log 2) values indicate an upregulation and negative (log 2) values a downregulation of the relevant gene.

(233) The deregulated genes for skin samples treated with 0.5 M concentration is used for further analysis.

(234) The experiment contains all up- and downregulated genes with the specified threshold.

(235) Data Analysis

(236) GeneGO's Metacore (Pathways and Maps) software.

(237) Experiments Analyses, Automatically Calculated Using GeneGO

(238) Experiment analysis consists of matching gene IDs for the sets of the uploaded files with gene IDs in functional ontologies in MetaCore. The ontologies include canonical pathway maps, GeneGo cellular processes, GO cellular processes and diseases categories. The degree of relevance to different categories for the uploaded datasets is defined by p-values, so that the lower p-value gets higher priority.

(239) Effects of CPPC 1 on Gene Expression Profile in Human Primary Epidermal Keratinocytes and Human Fibroblasts

(240) Cellular Process

(241) The cDNA microarray analysis reveales top ranked cellular and molecular processes in our interest as follows: anatomical structure development (p-value: 1.1217e-20), response to external stimulus (p-value: 7.9497e-20), positive regulation of cell proliferation (p-value 1.8286e-10), response to wounding (p-value: 4.1539e-14), cell proliferation (p-value: 1.1825e-08), regulation of cell proliferation (p-value: 1.4041e-13), response to extracellular stimulus (p-value: 2.4436e-09), skin development (p-value: 1.5864e-06), extracellular matrix organization and biogenesis (p-value: 1.1147e-06), extracellular structure organization (p-value: 1.8748e-06), regulation of epithelial cell proliferation (p-value: 5.9339e-06) and cell-matrix adhesion (p-value: 1.3430e-05).

Example 1, Positive Regulation of Cell Proliferation

(242) Positive regulation of cell proliferation is any process that activates or increases the rate or extent of cell proliferation.

(243) The GO/biological process categories are over-representative under the up- and downregulated genes after treatment with the substance CPPC 1. Following up- and downregulated genes play an important role in positive regulation of cell proliferation (Table 1).

(244) TABLE-US-00003 TABLE 1 Process: positive regulation of cell proliferation. Signal: the ratio as log2 value. Upregulated and downregulated genes. Process(es) genes - Positive regulation of cell proliferation # Gene Symbol Protein Protein name Signal P_value 263 TIMP1 TIMP1_HUMAN Metalloproteinase inhibitor 1 0.7312 0.0000029 138 IL6 IL6_HUMAN Interleukin-6 0.4854 0.00013 208 PTGS2 PGH2_HUMAN Prostaglandin G/H synthase 2 0.4222 0.021 256 TGFB1 TGFB1_HUMAN Transforming growth factor beta-1 0.3674 0.014 85 FGF7 FGF7_HUMAN Keratinocyte growth factor 0.3561 0.0012 60 DDR2 DDR2_HUMAN Discoidin domain-containing receptor 2 0.2869 0.0086 15 BCL2L1 BCLX_HUMAN Apoptosis regulator Bcl-X 0.263 0.017 20 C19orf10 CS010_HUMAN UPF0556 protein C19orf10 0.263 0.0022 32 CD81 CD81_HUMAN CD81 antigen 0.251 0.002 39 CDK2 CDK2_HUMAN Cell division protein kinase 2 0.2265 0.0046 200 PGF PLGF_HUMAN Placenta growth factor 0.2141 0.0058 40 CDK4 CDK4_HUMAN Cell division protein kinase 4 0.1375 0.034 245 STAT5B STA5B_HUMAN Signal transducer and activator of 0.1844 0.011 transcription 5B 71 EGF EGF_HUMAN Pro-epidermal growth factor 0.2345 0.044 269 TNFSF13B TN13B_HUMAN Tumor necrosis factor ligand superfamily 0.2688 0.0097 member 13B 140 IL7 IL7_HUMAN Interleukin-7 0.3585 0.0051 67 E2F3 E2F3_HUMAN Transcription factor E2F3 0.3959 0.0031 148 JUN JUN_HUMAN Transcription factor AP-1 0.5778 0.00000026
The Results of the Most Relevant Networks and Pathways Maps

(245) The cDNA microarray analysis of treated skin with CPPC 1 results many deregulated genes related to cell organization and communication, also mediated by integrins. The most relevant network objects based on network analysis and pathways maps (the top scored networks and maps) are the cell adhesion cell-matrix interactions (p-value: 4.204e-18), integrin-mediated cell adhesion and migration (p-Value: 1.697e-07), cell adhesion extracellular matrix remodeling map (p-value: 4.076e-16). The adhesion of cells to the extracellular matrix (ECM) is a dynamic process, mediated by a series of matrix-associated and cell-surface molecules that interact with each other in a spatially and temporally regulated manner. These interactions play a major role in tissue formation, cellular migration and the induction of adhesion-mediated transmembrane signals.

(246) The extra cellular matrix (ECM) is the extracellular part of tissue that usually provides structural support to the cells in addition to performing various important functions. Formation of the ECM is essential for processes like growth, wound healing, and fibrosis. The extracellular matrix is the defining feature of connective tissue. The ECM and cell adhesion and migration processes in fact plays at least three important roles relevant in cosmetics:

(247) i) Mechanical: tensile and compressive strength and elasticity.

(248) ii) Protection: buffering against extracellular change and retention of water.

(249) iii) Organization: control of cell behavior by binding of growth factors and interaction will cell-surface receptors.

(250) The ECM's main components are various glycoproteins like collagens, fibrin, elastin, fibronectins, laminins, and nidogens and other molecular compounds like proteoglycans and hyaluronic acid.

(251) Results for Cosmetic Applications

(252) The results of CPPC 1 in gene level gives insights into the potential action of the compound CPPC 1 as an integrin ligand in a variety of skin/hair regeneration, skin/hair care and/or disorders. Interactions with integrins and thus promotion of extracellular matrix proteins and a consecutive increased of matrix organization capacity, is believed to cause anti-wrinkle, anti-inflammation and promotion of hair growth in hair cycles and regarding to the hair follicle development.

(253) Thus, the outcome of cDNA data analysis is listed in Table 2 and 3 and is divided in two categories in cosmetics applications as follows:

(254) 1. Anti-wrinkles/Anti-ageing and anti-inflammation,

(255) 2. Hair growth/re-growthfollicle development.

Examples, Description of a Couple of the Deregulated Genes Obtained after the Treatment of Skin with CPPC 1

(256) S100 calcium binding protein A7, S100 calcium binding protein A8 and S100 calcium binding protein A9 (S100A7, S100A8 and S100A9) are downregulated genes after treatment of skin cells with CPPC 1.

(257) S100A7A is believed to be involved in epidermal differentiation and inflammation and thus is believed to be important for the pathogenesis of psoriasis and other diseases. The S100A7 protein, also known as psoriasin, has important functions as a mediator and regulator in skin differentiation and disease (psoriasis), in breast cancer, and as a chemotactic factor for inflammatory cells (Kulski et al., Journal of Molecular Evolution (2003), 56(4), 397-406).

(258) In addition, Lener et al. investigate genes involved in the natural aging process of the human skin. They found that in total 105 genes change their expression over 1.7-fold during the aging process in the human skin.

(259) S100A7 and S100A9 have been described as genes up-regulated in old skin (T. Lener et al., Experimental Gerontology 41 (2006), 387-397).

(260) S100 proteins are the largest subgroup of Ca.sup.2+ binding proteins with the EF-hand structural motif. A unique feature of this protein family is that individual members are localized in specific cellular compartments. For example, various S100 proteins are expressed in very restricted regions of the hair follicle (Kizawa K et al., Methods Mol Biol. (2005).

(261) Extracellular proteases are crucial regulators of cell function. The family of matrix metalloproteinases (MMPs) has classically been described in the context of extracellular matrix (ECM) remodeling, which occurs throughout life in diverse processes that range from tissue morphogenesis to wound healing. Recent evidence has implicated MMPs in the regulation of other functions, including survival, angiogenesis, inflammation and signaling.

(262) MMPs are secreted from keratinocytes and fibroblasts and break down collagen and other proteins that comprise the dermal extracellular matrix. Imperfect repair of the dermal damage impairs the functional and structural integrity of the extracellular matrix. Repeated sun exposure causes accumulation of dermal damage that eventually results in characteristic wrinkling of photodamaged skin (Gary J. Fisher et al., ARCH DERMATOL, vol. 138 (2002). In the skin, the primary role of MMP enzymes is to recycle skin matrix, particularly the structural proteins collagen and elastin. Reduces MMP activities degrade skin connective tissue and prevents loss of procollagen expression.

(263) Decreasing the expression or activity of matrix metalloproteases has an effect on the biological collagen catabolic process towards skin treatment of aging and psoriasis.

(264) The downregulated MMP genes after treatment of skin cells with CPPC 1 are MMP7, 13, 16, 23, 25. It is likely that downregulation of MMPs results in increase of collagen fibrils. Also laminin-alpha3, laminin-alpha4, lamninin-beta1, laminin-gamma1, collagen-alpha I (XV), collagen-alpha I (IV), and collagen-alpha II(IV) are significantly upregulated.

(265) Laminin is thought to mediate the attachment, migration, and organization of cells into tissues during embryonic development by interacting with other extracellular matrix components.

(266) Tissue inhibitor of metalloproteinases (TIMP-1) is strongly up-regulated after treatment of skin cells with CPPC 1. TIMP1 has been described as a cell survival factor. TIMP-1 is one representative of the natural MMP inhibitor family, encompassing four members. Its expression is decreased with fibroblast senescence, both ex vivo and in vivo, thus contributing to increased catabolic activity within dermis. TIMP-1 displays multiple biological functions. It inhibits most MMPs. Thus, CPPC 1 activates upregulation dermal fibroblast collagen production and downregulation of collagen degradation.

(267) Integrin-mediated cell adhesion and signaling events are essential for the proper development and homeostasis of most epithelial tissues.

(268) Ultraviolet (UV) irradiation from the sun reduces production of type I procollagen (COLI), the major structural protein in human skin. Photo-aging is the most common form of skin damage and is associated with skin carcinoma. UV irradiation inhibits TGF-beta1-induced type I procollagen gene expression in cultured human skin fibroblasts (Quan et al., AJP (2004), 165, No. 3, 741-751). TGF-beta is multifunctional protein that control proliferation, differentiation, and other functions in many cell types. TGF-beta/Smad pathway is the major regulator of type I procollagen synthesis in human skin.

(269) In the present gene expression experiment, TGF-beta1, TGF-beta3, and LTBP1 (latent TGF-beta binding protein) genes are upregulated after treatment of skin cells with CPPC 1.

(270) LTBP1 is one of the naturally occurring RGD ligands for v-integrins. Its role in activation and release of active TGF-beta has been described (Sheppard, Cancer and Metastasis Reviews 24 (2005), 395-402).

(271) Also, CTGF (connective tissue growth factor) is produced by skin fibroblasts after activation with TGF-beta.

(272) In addition, superoxide dismutase (SOD2), which catalyzes the dismution of superoxide into oxygen and hydrogen peroxide, is significantly upregulated. SOD is an important anti-oxidant defense in nearly all cells exposed to oxygen (skin/hair).

(273) Integrins are heterodimeric adhesion receptors composed of an alpha and beta subunits. Most integrins recognize several proteins of ECM, including laminin, fibronectin, and collagen (types I, II, and IV), elastin, fibulin, osteonectin, hyaluronic acid, and nidogen (Lee J W et al., Molecules and Cells (2004), 17(2):188-202). CPPC 1 is a highly active av-beta3/5 ligand and low active av-beta6 ligand (in vitro studies).

(274) It promotes cell proliferation, cell adhesion and extracellular structure organization by outcoming of cDNA microarray data and the published references.

(275) In skin and hair follicle biology, beta1 integrins and their ligands are of particular interest (Kloepper, J. E. Experimental Cell Research (2008). Integrin beta1 (ITGB1) shows a significant upregulation in our study.

(276) Integrin alpha5 and alpha3 (fibronectin receptor alpha, ITGA5 and ITGA3) are also upregulated in the skin treated by CPPC 1. It is known that integrin alpha5/beta1 mediates fibronectin-induced epithelial cell proliferation through activation of the EGFR. Fibronectins are proteins that connect cells with collagen fibers in the ECM, allowing cells to move through the ECM.

(277) TGF-beta superfamily of signaling molecules are involved in the regulation of many developmental processes that contain the interaction between mesenchymal and epithelial tissues. Smad7 is a potent inhibitor of many members of the TGF-beta family, notably TGF-beta and activin. Klopcic B et al. has reported that TGF-beta superfamily signaling is essential for development of hair, tooth, and T-cells as well as differentiation and proliferation control in adult tissues (Eur J, Cell Biol. (2007), 86(11-12):781-99).

(278) It is known that the inhibition of BMP signaling affects growth and differentiation in the anagen hair follicle. BMP stimulate differentiation of epidermal and hair follicle keratinocytes, inhibit initiation of hair growth, promote melanocyte proliferation and modulate melanogenesis, as well as an increase in calcitonin gene-related peptide expression in sensory neurons innervating skin. Downregulation of BMP-2 retards the entry in catagen phase in hair follicle cycling. Inhibition of BMP signaling also results in the generation of new intestinal stem cell proliferation (noggin is the natural inhibitor of BMP signalingL. M. Hogan et al., The EMBO Journal (2000)).

(279) BMP-2 suppresses proliferative activity and support diffentiation, while FGF-7 (fibroblast growth factor-7) induces anagen phase of hair follicle growth (Paus R., Physiol Rev (2001), 81:449-494).

(280) In the treated skin with CPPC 1, BMP-1, -2, -3, -7, and -10 are downregulated, while FGF-7 is significantly upregulated.

(281) Treatment with FGF-7 at concentrations of 10 ng/ml or greater significantly stimulates hair fiber elongation in human scalp hair follicle organ cultures (Iino M et al., Journal of Investigative Dermatology (2007)). Thus, stimulating of fibroblast growth factor-7 gene expression is believed stimulate hair growth.

(282) Fibulin-1 and -2 are extracellular matrix proteins, which belongs to the RGD class of proteins with unique structural features. A locally restricted expression pattern of fibulin-1 and fibulin-2 mRNA and protein at sites of epithelial-mesenchymal interactions was detected in two tissues, the developing tooth and hair follicles (Hang H Y et al., Dev Dyn (1996), 205(3):348-64).

(283) CPPC 1 is believed to bind at the integrin RGD site like fibulin and thus is believed promote the upregulation of fibulin and other extracellular matrix-RGD-proteins like fibronectin, procollagen, laminin, etc.

(284) DNA topoisomerase (TOP) are a family of enzymes that are involved in DNA replication and metabolism. The enzymes tie or untie DNA knots so that the DNA can replicate effectively. They regulate the helical structure of the double-stranded DNA by breaking one (topoisomerase type I) or both (topoisomerase type II) strands of the DNA helix (Wang J., Adv. Pharmacol. (1994), 29A:1-19). TOP-2 belongs to the genes, which are downregulated in the human anagen hair follicle bulge (Manabu Ohyama et al., J. Clin. Invest. (2006), 116:249-260). Hair loss is known as the side effect of topoismerase inhibitors.

(285) From the gene expression analysis and due to the literature (Ludbrook S B et al., Biochemical Journal (2003)), it is known that integrins regulates TGF superfamily signaling, in our study both, TGF-beta1 and TGF-beta3 are upregulated. TGF-beta1 and -beta3 is believed to lead via different transcription factors (SP1, SMADs, AP-1, Wnt, SR1, OASIS, etc.) to the regulation of targets like bone morphogenic proteins (BMPs: downregulated), nidogen (NID: upregulated), fibulins (upregulated), antigen Ki-67 (upregulated), and TOP2A (upregulated). In addition TGF-beta/BMP signaling pathway is known for the regulation of hair follicle development and cycling (Kobielak, K. et al., J. Cell Biol. (2003), 163:609-623)

APPENDIX, TABLE 2 AND 3

(286) TABLE-US-00004 TABLE 2 Anti-ageing and anti-inflammation related deregulated genes by treated skin with CPPC 1. Compound Gene vs. Control, ratio ID Gene-Name UniProt RefSeq Log(2) value P-value 5 IL2 P60568 P01585 NM_000586 0.577766999 0.051 Q13169 16 IL7 P13232 NM_000880 0.358453971 0.0051 32 TXLN Q66K62 Q86T54 NM_175852 0.836501268 0.049 Q86T85 Q86T86 Q86Y86 Q86YW3 P40222 Q8N2Y3 37 IL17A Q16552 NM_002190 0.666576266 0.00097 41 TNF P01375 O43647 NM_000594 0.304006187 0.06 Q9P1Q2 Q9UIV3 83 TNFSF13B Q9Y275 NM_006573 0.268816758 0.0097 99 TNFSF10 P50591 NM_003810 0.268816758 0.012 105 TNFRSF25 Q93038 Q93036 NM_003790 0.268816758 0.00065 Q93037 Q92983 NM_148965 P78515 Q99831 NM_148966 Q99722 P78507 NM_148967 Q99830 O NM_148970 251 TNFRSF1B P20333 Q6YI29 NM_001066 0.358453971 0.036 Q16042 Q9UIH1 2344 TIMP1 P01033 Q14252 NM_003254 0.731183242 0.0000029 Q9UCU1 4085 SOCS1 O15524 O15097 NM_003745 0.333423734 0.000095 Q9NSA7 7972 S100A8 P05109 Q9UC92 NM_002964 0.49410907 0.000037 Q9UCJ0 7975 S100A9 P06702 Q9NYM0 NM_002965 0.415037499 0.0000036 Q9UCJ1 9156 S100A7 P31151 Q9H1E2 NM_002963 0.514573173 0.00004 37885 TNFC Q06643 P78370 NM_002341 0.340075442 0.02 Q99761 NM_009588 18 IL8 Q9C077 P10145 NM_000584 0.321928095 0.019 Q6FGF6 Q6LAE6 Q96RG6 55 TUBB P07437 NM_178014 0.659924558 0.0005 499 ITGB1 P05556 P78466 NM_002211 0.378511623 0.0014 P78467 Q13089 NM_033666 Q14647 Q13090 NM_033667 Q13212 Q13091 NM_033668 Q14622 NM_033669 NM_133376 552 ITGA3 P26006 NM_002204 0.344828497 0.0013 NM_005501 556 ITGA5 P08648 Q96HA5 NM_002205 0.378511623 0.000067 1221 KI67 P46013 Q5VWH2 NM_002417 0.310340121 0.011 2275 COL15A1 P39059 Q5T6J4 NM_001855 0.367371066 0.011 Q9Y4W4 2289 COL4A1 P02462 Q9NYC5 NM_001845 0.50589093 0.0000005 2305 COL4A2 P08572 NM_001846 0.321928095 0.00022 2344 TIMP1 P01033 Q14252 NM_003254 0.731183242 0.0000029 Q9UCU1 2364 LAMA3 Q96TG0 Q16787 NM_000227 0.298658316 0.045 Q13679 Q13680 NM_198129 2366 LAMA4 Q9UE18 Q9UJN9 NM_002290 0.604071324 0.00000058 Q16363 Q15335 Q14735 Q14731 Q4LE44 Q5SZG8 2370 LAMB1 P07942 NM_002291 0.321928095 0.0036 2377 LAMG1 P11047 NM_002293 0.321928095 0.00007 2493 MMP13 P45452 NM_002427 0.473931188 0.0089 2505 MMP7 P09237 Q9BTK9 NM_002423 0.395928676 0.001 2533 SPARC P09486 NM_003118 0.367371066 0.025 4749 ELA2 P08246 P09649 NM_001972 0.234465254 0.073 Q6B0D9 Q6LDP5 5239 MMP23A- Q9UBR9 O75900 NM_006983 0.64385619 0.00000059 MMP23B O75894 O75895 NR_002946 Q5QPQ8 Q76P96 Q7LDM6 Q7LDM7 Q9UJK8 O 6926 LCN2 P80188 P30150 NM_005564 0.556393349 0.0000004 Q92683 7972 S100A8 P05109 Q9UC92 NM_002964 0.49410907 0.000037 Q9UCJ0 7975 S100A9 P06702 Q9NYM0 NM_002965 0.415037499 0.0000036 Q9UCJ1 9156 S100A7 P31151 Q9H1E2 NM_002963 0.514573173 0.00004 11207 KRT9 P35527 O00109 NM_000226 0.268816758 0.012 Q14665 20743 MMP25 Q9NPA2 Q9H3Q0 NM_022468 0.268816758 0.093 NM_022718 30439 TGFB1 P01137 Q9UCG4 NM_000660 0.367371066 0.014 30442 TGFB3 P10600 NM_003239 0.321928095 0.027 35735 MMP16_2 P51512 Q14824 NM_022564 0.340075442 0.036 Q52H48 38095 SPRR1A P35321 Q9UDG4 NM_005987 0.304006187 0.00069 38182 SOD2 P04179 P78434 NM_000636 0.298658316 0.005 Q16792 Q96EE6 NM_001024465 Q9P2Z3 Q5TCM1 NM_001024466

(287) TABLE-US-00005 TABLE 3 Hair relevant deregulated genes by treated skin with CPPC 1 Compound Gene vs. Control, ratio ID Gene-Name UniProt RefSeq Log(2) value P-value 2356 BMP7 Q9NTQ7 Q9H512 NM_001719 0.888968688 0.0019 P18075 9302 BMP10 O95393 NM_014482 0.415037499 0.015 38095 SPRR1A P35321 Q9UDG4 NM_005987 0.304006187 0.00069 3151 ASC Q9HBD0 Q9NXJ8 NM_145183 0.304006187 0.012 Q9BSZ5 Q9ULZ3 Q96D12 35633 CASP8 Q14790 O14676 NM_001228 0.304006187 0.016 Q14791 Q14792 NM_033355 Q14793 Q14794 NM_033356 Q14795 Q14796 NM_033358 Q15780 Q 2104 GDF10 Q9UCX6 P55107 NM_004962 0.286304185 0.044 7969 S100A12 P80511 P83219 NM_005621 0.251538767 0.019 1282 BMP2 P12643 NM_001200 0.251538767 0.0072 35624 BMP1_2 P13497 Q13292 NM_001199 0.234465254 0.098 Q13872 Q14874 NM_006128 Q99421 Q99422 NM_006129 Q99423 Q9UL38 P46721 Q 1221 KI67 P46013 Q5VWH2 NM_002417 0.310340121 0.011 4091 STAT1 P42224 NM_007315 0.310340121 0.0014 NM_139266 30442 TGFB3 P10600 NM_003239 0.321928095 0.027 11184 KRT19 P08727 Q5XG83 NM_002276 0.333423734 0.032 Q6NW33 Q7L5M9 Q96A53 Q96FV1 Q9BYF9 Q9P1Y4 552 ITGA3 P26006 NM_002204 0.344828497 0.0013 NM_005501 5203 FGF7 P21781 NM_002009 0.35614381 0.0012 30439 TGFB1 P01137 Q9UCG4 NM_000660 0.367371066 0.014 281 MCL1 Q07820 Q9UNJ1 NM_021960 0.367371066 0.0069 Q9NRQ3 Q9NRQ4 NM_182763 Q9HD91 Q9UHR7 Q9UHR8 Q9UHR9 2533 SPARC P09486 NM_003118 0.367371066 0.025 499 ITGB1 P05556 P78466 NM_002211 0.378511623 0.0014 P78467 Q13089 NM_033666 Q14647 Q13090 NM_033667 Q13212 Q13091 NM_033668 Q14622 NM_033669 NM_133376 556 ITGA5 P08648 Q96HA5 NM_002205 0.378511623 0.000067 2457 FBLN2 P98095 NM_001004019 0.378511623 0.00029 NM_001998 1613 SDCBP O00173 O00560 NM_005625 0.389566812 0.000026 O43391 11190 KRT2A P35908 NM_000423 0.40053793 0.072 2437 QSOX1 O00391 Q13876 NM_001004128 0.432959407 0.0015 Q59G29 Q5T2X0 NM_002826 Q8WVP4 Q8TDL6 7586 CLU P10909 P11380 NM_001831 0.443606651 0.0048 P11381 Q7Z5B9 NM_203339 11154 FLOT2 Q14254 NM_004475 0.454175893 0.0052 118 CCNB2 O95067 NM_004701 0.464668267 0.000027 2515 NIDOGEN: P14543 Q14942 NM_002508 0.475084883 0.000093 (NID) Q59FL2 Q5TAF2 Q5TAF3 Q86XD7 2483 ADAM9 Q8NFM6 Q10718 NM_001005845 0.485426827 0.0051 Q13443 NM_003816 17158 IGFBP4 P22692 NM_001552 0.485426827 0.00089 5177 FGF1 P05230 P07502 NM_000800 0.50589093 0.0016 NM_033136 NM_033137 21796 FBLN1_4 Q8TBH8 Q9HBQ5 NM_001996 0.516015147 0.000016 Q9UH41 P23142 NM_006485 P23143 P23144 NM_006486 P37888 Q9UGR4 NM_006487 Q5TIC4 Q 10509 SEPRASE Q12884 O00199 NM_004460 0.545968369 0.00000071 Q86Z29 Q99998 Q9UID4 2465 LTBP1 P22064 Q14766 NM_000627 0.555816155 0.000065 Q8TD95 NM_206943 9410 TOP2A Q9UP44 Q9UQP9 NM_001067 0.622930351 0.000024 P11388 Q9HB24 Q9HB25 Q9HB26 Q71UN1 Q71UQ5

Example 13: Compositions

(288) Formulations for compositions, preferably preferably non-therapeutic compositions, cosmetic compositions and/or topical compositions, comprising compounds selected from the cyclic peptides according to the invention are shown by way of example below. The INCI names of the commercially available compounds are also shown.

(289) UV-Pearl, OMC stands for the composition having the INCI name: Water (for EU: Aqua), Ethylhexyl Methoxycinnamate, Silica, PVP, Chlorphenesin, BHT; this composition is commercially available under the name EusolexUV Pearl OMC from Merck KGaA, Darmstadt.

(290) The other UV Pearl products indicated in the tables are each of analogous composition with OMC replaced by the UV filter indicated.

(291) TABLE-US-00006 TABLE 1a W/O emulsions (data in % by weight) 1-1 1-2 1-3 1-4 1-5 1-6 1-7 1-8 1-9 1-10 Titanium Dioxide 2 5 3 Cyclo-(Arg-Gly-Asp-DPhe- 0.01 0.005 0.001 0.0005 0.005 0.005 0.005 Acha) Cyclo-(Arg-Gly-Asp-DPhe- 0.0005 0.01 0.005 (NMe)Val) Zinc Oxide 5 2 UV-Pearl, OMC 30 15 15 15 15 15 15 15 15 15 Polyglyceryl-3-Dimerate 3 3 3 3 3 3 3 3 3 3 Cera Alba 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 Hydrogenated Castor Oil 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Paraffinium Liquidum 7 7 7 7 7 7 7 7 7 7 Caprylic/Capric Triglyceride 7 7 7 7 7 7 7 7 7 7 Hexyl Laurate 4 4 4 4 4 4 4 4 4 4 PVP/Eicosene Copolymer 2 2 2 2 2 2 2 2 2 2 Propylene Glycol 4 4 4 4 4 4 4 4 4 4 Magnesium Sulfate 0.6 0.6 0.6 0.6 0.6 0.6 0.6 0.6 0.6 0.6 Tocopherol 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Tocopheryl Acetate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Cyclomethicone 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Propylparaben 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 Methylparaben 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 Water to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100

(292) TABLE-US-00007 TABLE 1b 1-11 1-12 1-13 1-14 1-15 1-16 1-17 1-18 Titanium Dioxide 3 2 3 2 5 Benzylidene Malonate Polysiloxane 1 0.5 Methylene Bis-benzotriazolyl 1 1 0.5 Tetramethylbutylphenol Cyclo-(Arg-Gly-Asp-DPhe-Acha) 0.01 0.005 0.001 0.0005 0.005 Cyclo-(Arg-Gly-Asp-DPhe- 0.0005 0.01 0.005 (NMe)Val) Polyglyceryl-3-Dimerate 3 3 3 3 Cera Alba 0.3 0.3 0.3 0.3 2 2 2 2 Hydrogenated Castor Oil 0.2 0.2 0.2 0.2 Paraffinium Liquidum 7 7 7 7 Caprylic/Capric Triglyceride 7 7 7 7 Hexyl Laurate 4 4 4 4 PVP/Eicosene Copolymer 2 2 2 2 Propylene Glycol 4 4 4 4 Magnesium Sulfate 0.6 0.6 0.6 0.6 Tocopherol 0.5 0.5 0.5 0.5 Tocopheryl Acetate 0.5 0.5 0.5 0.5 1 1 1 1 Cyclomethicone 0.5 0.5 0.5 0.5 Propylparaben 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 Methylparaben 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 Dicocoyl Pentyerythrityl Citrate (and) 6 6 6 6 Sorbitan Sesquioleate (and) Cera Alba (and) Aluminium Stearate PEG-7 Hydrogenated Castor Oil 1 1 1 1 Zinc Stearate 2 2 2 2 Oleyl Erucate 6 6 6 6 Decyl Oleate 6 6 6 6 Dimethicone 5 5 5 5 Tromethamine 1 1 1 1 Glycerine 5 5 5 5 Allantoin 0.2 0.2 0.2 0.2 Water to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100

(293) TABLE-US-00008 TABLE 1c 1-19 1-20 1-21 1-22 1-23 1-24 1-25 1-26 1-27 1-28 1-29 Titanium Dioxide 2 5 3 3 Benzylidene Malonate 1 1 1 Polysiloxane Cyclo-(Arg-Gly-Asp-DPhe-Acha) 0.01 0.005 0.001 0.0005 0.005 0.005 0.001 0.0005 Cyclo-(Arg-Gly-Asp-DPhe- 0.0005 0.01 0.005 (NMe)Val) Zinc Oxide 5 2 UV-Pearl, OCR 10 5 UV-Pearl, 10 EthylhexylDimethylPABA UV-Pearl, Homosalate 10 UV-Pearl, Ethylhexyl Salicylate 10 UV-Pearl, OMC. BP-3 10 UV-Pearl, OCR. BP-3 10 UV-Pearl, Ethylhexyl Dimethyl 10 PABA, BP-3 UV-Pearl, Homosalate, BP-3 10 UV-Pearl, Ethylhexyl Salicylate, 10 BP-3 BMDBM 2 UV-Pearl, OMC, 25 4-Methylbenzylidene Camphor Polyglyceryl-3-Dimerate 3 3 3 3 3 3 3 3 3 3 3 Cera Alba 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 Hydrogenated Castor Oil 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Paraffinium Liquidum 7 7 7 7 7 7 7 7 7 7 7 Caprylic/Capric Triglyceride 7 7 7 7 7 7 7 7 7 7 7 Hexyl Laurate 4 4 4 4 4 4 4 4 4 4 4 PVP/Eicosene Copolymer 2 2 2 2 2 2 2 2 2 2 2 Propylene Glycol 4 4 4 4 4 4 4 4 4 4 4 Magnesium Sulfate 0.6 0.6 0.6 0.6 0.6 0.6 0.6 0.6 0.6 0.6 0.6 Tocopherol 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Tocopheryl Acetate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Cyclomethicone 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Propylparaben 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 Methylparaben 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 Water to 100

(294) TABLE-US-00009 TABLE 2a O/W emulsions, data in % by weight 2-1 2-2 2-3 2-4 2-5 2-6 2-7 2-8 2-9 2-10 Titanium Dioxide 2 5 3 Methylene Bis-benzotriazolyl 1 2 1 Tetramethylbutylphenol Cyclo-(Arg-Gly-Asp-DPhe-Acha) 0.01 0.005 0.001 0.0005 0.005 0.005 0.001 Cyclo-(Arg-Gly-Asp-DPhe- 0.0005 0.01 0.005 (NMe)Val) Ectoine 1 3 5 1 2 4-Methylbenzylidene Camphor 2 3 4 3 2 BMDBM 1 3 3 3 3 3 3 Stearyl Alcohol (and) Steareth-7 3 3 3 3 3 3 3 3 3 3 (and) Steareth-10 Glyceryl Stearate (and) Ceteth-20 3 3 3 3 3 3 3 3 3 3 Glyceryl Stearate 3 3 3 3 3 3 3 3 3 3 Microwax 1 1 1 1 1 1 1 1 1 1 Cetearyl Octanoate 11.5 11.5 11.5 11.5 11.5 11.5 11.5 11.5 11.5 11.5 Caprylic/Capric Triglyceride 6 6 6 6 6 6 6 6 6 6 Oleyl Oleate 6 6 6 6 6 6 6 6 6 6 Propylene Glycol 4 4 4 4 4 4 4 4 4 4 Glyceryl Stearate SE Stearic Acid Persea Gratissima Propylparaben 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 Methylparaben 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 Tromethamine 1.8 Glycerine Water to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100

(295) TABLE-US-00010 TABLE 2b 2-11 2-12 2-13 2-14 2-15 2-16 2-17 2-18 Titanium Dioxide 3 2 2 5 Benzylidene Malonate 1 0.5 Polysiloxane Cyclo-(Arg-Gly-Asp-DPhe-Acha) 0.01 0.005 0.001 0.0005 0.005 Cyclo-(Arg-Gly-Asp-DPhe- 0.0005 0.01 0.005 (NMe)Val) Ectoine 0.5 1 3 1 Zinc Oxide 2 UV-Pearl, OMC 15 15 15 30 30 30 15 15 4-Methylbenzylidene Camphor 3 BMDBM 1 Phenylbenzimidazole Sulfonic Acid 4 Stearyl Alcohol (and) Steareth-7 3 3 3 3 (and) Steareth-10 Glyceryl Stearate (and) Ceteth-20 3 3 3 3 Glyceryl Stearate 3 3 3 3 Microwax 1 1 1 1 Cetearyl Octanoate 11.5 11.5 11.5 11.5 Caprylic/Capric Triglyceride 6 6 6 6 14 14 14 14 Oleyl Oleate 6 6 6 6 Propylene Glycol 4 4 4 4 Glyceryl Stearate SE 6 6 6 6 Stearic Acid 2 2 2 2 Persea Gratissima 8 8 8 8 Propylparaben 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 Methylparaben 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 Tromethamine 1.8 Glycerine 3 3 3 3 Water to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100

(296) TABLE-US-00011 TABLE 2c 2-19 2-20 2-21 2-22 2-23 2-24 2-25 2-26 2-27 2-28 Titanium Dioxide 3 3 2 Cyclo-(Arg-Gly-Asp-DPhe- 0.01 0.005 0.001 0.0005 0.005 0.005 0.001 Acha) Cyclo-(Arg-Gly-Asp-DPhe- 0.0005 0.01 0.005 (NMe)Val) Zinc Oxide 5 2 2 UV-Pearl, OMC 15 15 15 15 15 15 15 15 15 15 Caprylic/Capric Triglyceride 14 14 14 14 14 14 14 14 14 14 Oleyl Oleate Propylene Glycol Glyceryl Stearate SE 6 6 6 6 6 6 6 6 6 6 Stearic Acid 2 2 2 2 2 2 2 2 2 2 Persea Gratissima 8 8 8 8 8 8 8 8 8 8 Propylparaben 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 Methylparaben 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 Glyceryl Stearate. Ceteareth-20. Ceteareth-10. Cetearyl Alcohol. Cetyl Palmitate Ceteareth-30 Dicaprylyl Ether Hexyldecanol, Hexyldexyl Laurate Cocoglycerides Tromethamine Glycerine 3 3 3 3 3 3 3 3 3 3 Water to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100

(297) TABLE-US-00012 TABLE 3 Gels, data in % by weight 3-1 3-2 3-3 3-4 3-5 3-6 3-7 3-8 3-9 3-10 Titanium Dioxide 2 5 3 Cyclo-(Arg-Gly-Asp-DPhe-Acha) 0.01 0.005 0.001 0.0005 0.005 0.005 0.001 Cyclo-(Arg-Gly-Asp-DPhe- 0.0005 0.01 0.005 (NMe)Val) Benzylidene Malonate 1 1 2 1 1 Polysiloxane Methylene Bis-benzotriazolyl 1 1 2 1 Tetramethylbutylphenol Zinc Oxide 2 5 2 UV-Pearl, Ethylhexyl 30 15 15 15 15 15 15 15 15 15 Methoxycinnamate 4-Methylbenzylidene Camphor 2 Butylmethoxydibenzoylmethane 1 Phenylbenzimidazole Sulfonic Acid 4 Prunus Dulcis 5 5 5 5 5 5 5 5 5 5 Tocopheryl Acetate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Caprylic/Capric Triglyceride 3 3 3 3 3 3 3 3 3 3 Octyldodecanol 2 2 2 2 2 2 2 2 2 2 Decyl Oleate 2 2 2 2 2 2 2 2 2 2 PEG-8 (and) Tocopherol (and) 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 Ascorbyl Palmitate (and) Ascorbic Acid (and) Citric Acid Sorbitol 4 4 4 4 4 4 4 4 4 4 Polyacrylamide (and) C13-14 3 3 3 3 3 3 3 3 3 3 Isoparaffin (and) Laureth-7 Propylparaben 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 Methylparaben 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 Tromethamine 1.8 Water to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 a = aqueous gel

(298) TABLE-US-00013 TABLE 2d O/W emulsions, data in % by weight 2-19 2-20 2-21 2-22 2-23 2-24 2-25 2-26 2-27 2-28 Titanium dioxide 3 3 2 Cyclo-(Arg-Gly-Asp-DPhe- 0.01 0.005 0.001 0.0005 0.005 0.005 0.001 Acha) Cyclo-(Arg-Gly-Asp-DPhe- 0.0005 0.01 0.005 (NMe)Val) Ectoine 1 3 5 1 2 Methylene Bis-Benztriazolyl 1 2 1 1 1 0.5 Tetramethylbutylphenol Zinc oxide 5 2 2 UV-Pearl, OMC 15 15 15 15 15 15 15 15 15 15 Caprylic/Capric Triglyceride 14 14 14 14 14 14 14 14 14 14 Oleyl Oleate Propylene Glycol Glyceryl Stearate SE 6 6 6 6 6 6 6 6 6 6 Stearic Acid 2 2 2 2 2 2 2 2 2 2 Persea Gratissima 8 8 8 8 8 8 8 8 8 8 Propylparabene 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 Methylparabene 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 Glyceryl Stearate, Ceteareth-20, Ceteareth-10, Cetearyl Alcohol, Cetyl Palmitate Ceteareth-30 Dicaprylyl Ether Hexyldecanol, Hexyldexyllaurate Cocoglycerides Tromethamine Glycerin 3 3 3 3 3 3 3 3 3 3 Water ad 100 ad 100 ad 100 ad 100 ad 100 ad 100 ad 100 ad 100 ad 100 ad 100

(299) TABLE-US-00014 TABLE 3 Gels, data in % by weight 3-1 3-2 3-3 3-4 3-5 3-6 3-7 3-8 3-9 3-10 Titanium dioxide 2 5 3 Cyclo-(Arg-Gly-Asp-DPhe-Acha) 0.01 0.005 0.001 0.0005 0.005 0.005 0.001 Cyclo-(Arg-Gly-Asp-DPhe- 0.0005 0.01 0.005 (NMe)Val) Benzylidene malonate 1 1 2 1 1 polysiloxane Methylene Bis-Benztriazolyl 1 1 2 1 Tetramethylbutylphenol Zinc oxide 2 5 2 UV-Pearl, Ethylhexyl 30 15 15 15 15 15 15 15 15 15 Mehtoxycinnamat 4-Methylbenzylidene Camphor 2 Butylmethoxydibenzoylmethane 1 Phenylbenzimidazole Sulfonic Acid 4 Prunus Dulcis 5 5 5 5 5 5 5 5 5 5 Tocopheryl Acetate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Caprylic/Capric Triglyceride 3 3 3 3 3 3 3 3 3 3 Octyldodecanol 2 2 2 2 2 2 2 2 2 2 Decyl Oleate 2 2 2 2 2 2 2 2 2 2 PEG-8 (and) Tocopherol (and) 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 Ascorbyl Palmitate (and) Ascorbic Acid (and) Citric Acid Sorbitol 4 4 4 4 4 4 4 4 4 4 Polyacrylamide (and) C13-14 3 3 3 3 3 3 3 3 3 3 Isoparaffin (and) Laureth-7 Propylparabene 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 Methylparabene 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 Tromethamine 1.8 Water ad 100 ad 100 ad 100 ad 100 ad 100 ad 100 ad 100 ad 100 ad 100 ad 100 a = aqueous gel

Example 14: Anti-Cellulite Compositions

(300) TABLE-US-00015 Components % Composition 1 Phase A Cetyl alcohol 2 Glyceryl Stearate 5 Caprylic/Capric 8 Triglyceride Isopropyl Palmitate 9 Phase B Glycerol 3 Preservatives 0.8 (Germaben II) Cyclo-(Arg-Gly- 0.005 Asp-DPhe-Acha) Water, demineralised ad 100 composition 2 Phase A Cetyl alcohol 2 Glyceryl Stearate 5 Caprylic/Capric 8 Triglyceride Isopropyl Palmitate 9 Phase B Glycerol 3 Preservatives 0.8 (Germaben II) Cyclo-(Arg-Gly- 0.0005 Asp-DPhe-Acha) Water, demineralised ad 100
Method:

(301) Method: Phases A and B are heated to 65-70 C. Give phase Phase B to Phase A without stirring. Homogenise and allow the mixture to cool to room temperature.

Example 15: Hair Care Formulations

(302) Cyclopeptide or CPPC 1 is Cyclo-(Arg-Gly-Asp-DPhe-Acha);

(303) CPPL 1 and CPPL 2 are liposomes with following composition:

(304) CPPL 1:

(305) CPPC 1 (0.01%) Ethanol (17.00%) Lecithin (5.00%) Water (ad 100)
CPPL 2: CPPC 1 (0.01%) Ethanol (17.00%) Ectoin (5.00%) Lecithin (5.00%) Water (ad 100)
1. Conditioning Shampoo

(306) TABLE-US-00016 Formula 1 Ingredients/INCI Declaration (Tradename) w/w % CPPC 1 0.001-0.01 UREA, DISODIUM PHOSPHATE, BIOTIN, 1.0 CITRIC Acid (RonaCare Biotin Plus) NIACINAMIDE 0.1 HYDROXYPROPYL GUAR 0.2 SODIUM COCOAMPHOACETATE 10 (Miranol Ultra C-32) AQUA (WATER), SODIUM LAURETH 32 SULFATE (Texapon NSO) PANTHENOL 0.5 SODIUM CHLORIDE 1.0 PARFUM q.s. CITRIC ACID q.s AQUA (WATER) ad 100
2. Anti-Dandruff Shampoo

(307) TABLE-US-00017 Formula 1 Ingredients/INCI Declaration (Tradename) w/w % CPPC 1 0.001-0.01 MICA, CI 77891 (Timiron Diamond Cluster MP-149 0.05 XANTHAN GUM 0.7 AQUA, SODIUM LAURETH SULFATE (Texapon NSO) PIROCTONE OLAMINE 0.50 COCAMIDOPROPYL BETAINE 5.00 PROPYLENE GLYCOL, 5-BROMO-5-NITRO-1,3- 0.20 DIOXANE (Bronidox L) PARFUM 0.20 AQUA (WATER) ad 100
3. Hair Growth Tonic

(308) TABLE-US-00018 Ingredients/INCI Declaration Formula 1 Formula 2 Formula 3 (Tradename) w/w % w/w % w/w % CPPC 1 0.001-0.01 CPPL 1 2.00-10.00 CPPL 2 2.00-10.00 CAFFEINE 0.50 0.50 0.50 ETHOXYDIGLYCOL, 3.00 3.00 3.00 PROPYLENE GLYCOL, BUTYLENE GLYCOL, SODIUM BENZOATE, POTASSIUM SORBATE (Exptapon Birke Spezial) PANTHENOL 0.40 0.40 0.40 ALCOHOL 10.00 10.00 10.00 TOCOPHERYL ACETATE 0.30 0.30 0.30 MENTHOL 0.10 0.10 0.10 PEG-40 HYDROGENATED 1.50 1.50 1.50 CASTOR OIL AQUA (WATER) ad 100 ad 100 ad 100
3. Vitamin Hair Growth Tonic

(309) TABLE-US-00019 Ingredients/INCI Declaration Formula 1 Formula 2 Formula 3 (Tradename) w/w % w/w % w/w % CPPC 1 0.001-0.01 CPPL 1 2.00-10.00 CPPL 2 2.00-10.00 NIACINAMIDE 2.00 2.00 2.00 BIOTIN 0.02 0.02 0.02 SALICYLIC ACID 0.10 0.10 0.10 ISOPROPYL ALCOHOL 10.00 10.00 10.00 PARFUM 0.05 0.05 0.05 AQUA (WATER) ad 100 ad 100 ad 100
4. Scalp Treatment Emulsion for Hair Growth

(310) TABLE-US-00020 Ingredients/INCI Declaration Formula 1 Formula 2 Formula 3 (Tradename) w/w % w/w % w/w % CPPC 1 0.001-0.01 CPPL 1 2.00-10.00 CPPL 2 2.00-10.00 ISOQUERCETIN 0.2 0.2 0.2 PROPYLENE GLYCOL 5.00 5.00 5.00 ACRYLATES/C10-30 ALKYL 0.20 0.20 0.20 ACRYLATE, CROSSPOLYMER SUCROSE STEARATE 1.00 1.00 1.00 DECYL OLEATE 3.00 3.00 3.00 DIMETHICONE 4.00 4.00 4.00 SODIUM HYDROXIDE 0.03 0.03 0.03 AQUA (WATER) ad 100 ad 100 ad 100
5. Scalp Treatment Emulsion

(311) TABLE-US-00021 Ingredients/INCI Declaration Formula 1 Formula 2 Formula 3 (Tradename) w/w % w/w % w/w % CPPC 1 0.001-0.01 CPPL 1 2.00-10.00 CPPL 2 2.00-10.00 CETEARYL ALCOHOL 2.50 2.50 2.50 CETEARETH-20 1.00 1.00 1.00 DISODIUM RUTINYL 1.00 1.00 1.00 DISULPHATE OLEYL ERUCATE 1.00 1.00 1.00 PROPYLPARABEN 0.05 0.05 0.05 METHYLPARABEN 0.15 0.15 0.15 AQUA (WATER) ad 100 ad 100 ad 100
6. Hair Conditioner

(312) TABLE-US-00022 Ingredients/INCI Declaration Formula 1 Formula 2 Formula 3 (Tradename) w/w % w/w % w/w % CPPC 1 0.001-0.01 CPPL 1 2.00-10.00 CPPL 2 2.00-10.00 CETEARYL ALCOHOL, 4.50 4.50 4.50 BEHENTRIMONIUM CHLORIDE (Incroquat Behenyl TMC) CYCLOPENTASILOXANE, 4.00 4.00 4.00 CYCLOHEXASILOXANE PROPYLENE GLYCOL, 0.70 0.70 0.70 DIAZOLIDINYL UREA, METHYLPARABEN, PROPYLPARABEN (Germaben II) AQUA (WATER) ad 100 ad 100 ad 100
7. Hair Spray, Aerosol

(313) TABLE-US-00023 Formula 1 Ingredients/INCI Declaration (Tradename) w/w % CPPC 1 0.001-0.01 Ectoin 1.00 BENZOPHENONE -3 0.50 PVP/VA/VINYL PROPIONATE 5.00 COPOLYMER PROPANE/BUTANE (40:60) 30.00 AQUA (WATER) 10.00 ETHANOL ad 100
8. Hair Styling Gel with UV Protection

(314) TABLE-US-00024 Ingredients/INCI Declaration Formula 1 Formula 2 Formula 3 (Tradename) w/w % w/w % w/w % CPPC 1 0.001-0.01 CPPL 1 2.00-10.00 CPPL 2 2.00-10.00 ACRYLATES/C10-30 ALKYL 0.50 0.50 0.50 ACRYLATE, CROSSPOLYMER (Carbopol Ultrez 21) ISOPROPYL ALCOHOL 15.00 15.00 15.00 PVP 1.50 1.50 1.50 PROPYLENE GLYCOL, 0.20 0.20 0.20 DIAZOLIDINYL UREA, METHYLPARABEN, PROPYLPARABEN AQUA, CETRIMONIUM 0.30 0.30 0.30 CHLORIDE AQUA (WATER), ETHYLHEXYL, 10.00 10.00 10.00 METHOXYCINNAMATE, SILICA, PVP, CHLORPHENESIN, BHT (Eusolex UV-Pearls 2292) AQUA (WATER) ad 100 ad 100 ad 100
9. Cold Wave Solution

(315) TABLE-US-00025 Formula 1 Ingredients/INCI Declaration (Tradename) w/w % CPPC 1 0.001-0.01 AMMONIUM THIOGLYCOLATE, AQUA 16.00 AMMONIUM BICARBONATE 0.50 PVP 2.00 POTASSIUM COCOYL HYDROLYZED 1.00 COLLAGEN NONOXYNOL-14 5.60 TOCOPHERYL ACETATE 1.00 AQUA (WATER) ad 100

Example 16: Improvement of Skin Smoothness and Reduction of Wrinkle Depth In Vivo

(316) Goal of the Study:

(317) Determination of the effect of Cyclopeptide Cyclo-(Arg-Gly-Asp-DPhe-Acha) (=CPPC 1) on skin topography in vivo by means of Primos. The topical application of test products occurs on a defined area on the inner forearm as well as the crows' feet (eye area).

(318) Design of the Study:

(319) TABLE-US-00026 Subjects: Number of individuals.: 20 (+1 reserve subject) Sex: female Age range: 37-63 years (average: 44.8) Test Area: Inner sides of forearms Crows'feet Test Parameters: 1. Determination of skin roughness by means of PRIMOS 5.6 (GFMetechnik GmbH, Teltow, Germany) 2. Determination of wrinkle depth by means of PRIMOS 5.6 (GFMetechnik GmbH, Teltow, Germany) Design of study: Day 0 Determination of the parameters in the test areas First test product application Day 14 Determination of the parameters 8-12 hours following the last daily test product application Day 28 Determination of the parameters 8-12 hours following the last daily test product application
Measurement of Skin Roughness:

(320) PRIMOS (Phase-Shifting rapid in vivo measurement of skin) is a non-contact measurement device, which allows for real-time three-dimensional in vivo measurement of the micro topography of human skin based on the technology of active image triangulation. The measurement head consist of a digital micromirror device as projection unit and a CCD-camera as recording unit, mounted onto an adjustable rack. For active image triangulation an intensity encoded point M is projected onto the surface under investigation. Its image on the surface is recorded by the CCD-camera from a specific angle. The point M is a function of parameters like intensity, triangulation angle between projection system and camera and some other inner respectively outer coordinates of the camera and projection plane. The height information of the structured surface is coded in the distorted intensity pattern, which is recorded. The resolution and accuracy depends on the optical and topographical characteristics of the measured surface and on the noise characteristics of the measurement system. For accurate in vivo measurement of human skin, depending on the measured part of the human body (inner forearm, forehead, eye zone), different parameters of effective wavelength and amplification factor should be used.

(321) To regard the differences of human skin and avoid undesired distortions by movements, the fast phase-shift technique was used for the measurement (phase width: 16 & 64 pixels). For each measurement, a minimum of 3 recordings were made and the clearest image without movement distortions or artefacts was selected for further processing.

(322) At the end of the study, distortions due to body hairs were digitally removed and the macro structure (calculated by polynomial approximation), i.e. the curvature of the entire test area, subtracted to allow a proper analysis of the microstructure, i.e. surface roughness.

(323) Skin roughness was then assessed by means of the parameter R.sub.Z (mean depth of roughness). To mitigate potential directional effects, the evaluation was conducted using the arithmetic average of R.sub.Z from 32 radial cuts. The mean depth of roughness is defined as:

(324) $R_Z=\frac{1}{n}\underset{i=1}{\overset{n}{.Math.}}{R}_{\mathrm{Zi}}$
where n is the number of equal segments into which the scan length I has been divided into an R.sub.D is the maximum peak to valley depth within each of the segments. In accordance with the German Standard Din 4768/1, R.sub.Z was calculated using 5 segments of equal length.
Measurement of Wrinkle Depth

(325) The crows' feet area is recorded as 3D topography using the PRIMOS system as outlined above. To accurately detect deeper structures different settings for the fast-shift were used (phase width: 16, 64 & 128 pixels). For each measurement, a minimum of 3 recordings were made and the clearest image without movement distortions or artefacts was selected for further processing. On follow-up visits, the original captured data was projected onto the skin of the volunteers to help in the relocation process of the test area. At the end of the study, distortions due to body hairs were digitally removed and the macro structure (calculated by polynomial approximation), i.e. the curvature of the entire test area, subtracted to allow a proper analysis of the microstructure, i.e. wrinkles and surface roughness. Wrinkle depth was then assessed by means of the parameter R.sub.Max that is defined as the maximum vertical distance from the highest peak to the lowest valley of five segments of equal length. To mitigate locational effects, the evaluation was conducted using the arithmetic average of R.sub.max from 50 parallel cuts.

(326) Performance of Test

(327) The subjects of this study were between 37-63 years of age (average 44.8). The subjects were instructed not to use any topical preparations on the test areas starting from seven days prior to testing and until the end of the test. For cleansing, water or a mild syndet was allowed only (whole study inclusive the run-in phase).

(328) Prior the first application of the test products, measurements were taken at clearly defined sites of the inner sides of the forearms (skin hydration, biomechanical properties, skin roughness) and in the crows' feet region (wrinkle depth). One area on the inner side of the forearms remained untreated and served as control. Further measurement was performed after 14 and 28 days of application 8-12 hours following the last daily application (adaptation time: 30 min, room temperature: 211 C., relative humidity: 505%). The subjects used the test products (approximately 2 mg/cm.sup.2) twice daily (in the morning and the evening) and in a manner corresponding as largely as possible to that to be practised by the future customer.

(329) TABLE-US-00027 TABLE 01 Composition of the liposome raw material (Liposome RM) Ingredient Amount [%] Ectoin 5.00 Ethanol, denat. 17.2 Lecithin 5.00 Water ad 100

(330) TABLE-US-00028 TABLE 02 Composition of the liposome raw material incl. CPPC 1 (Liposome RM containing 100 ppm/1000 ppm CPPC 1) Ingredient Amount [%] Ectoin 5.00 Ethanol, denat. 17.2 Lecithin 5.00 CPPC 1 100 ppm or 1000 ppm Water ad 100

(331) TABLE-US-00029 TABLE 03 Composition of the test formulations Name of testproduct Placebo RM-0 RM-100 RM-1000 Raw material INCI % w/w g g g g A Montanov 202 Arachidyl Alcohol (and) 3.00 30 30 30 30 Behenyl Alcohol (and) Arachidylglucoside) Tegosoft DEC Diethylhexyl Carbonate 4.00 40 40 40 40 Cetiol A Hexyl Laurate 8.00 80 80 80 80 B Water Aqua 76.00 800 760 760 760 1,2-Propanediol Propylen Glycol 3.00 30 30 30 30 C Sepigel 305 Polyacrylamide (and) 1.00 10 10 10 10 C-13-14 Isoparaffine (and) Laureth-7 D Liposome RM-0 Aqua (Water), 4.00 40 (without CPPC 1) Alcohol denat., Lecithin, Ectoin Liposome RM-100 Aqua (Water), 4.00 40 (containing 100 Alcohol denat., ppm CPPC 1) Lecithin, Ectoin Liposome RM-1000 Aqua (Water), 4.0 40 (containing 1000 Alcohol denat., ppm CPPC 1) Lecithin, Ectoin Germaben II Propylene Glycol (and) 1.00 10 10 10 10 Diazolidinyl Urea (and) Methylparaben (and) Propylparaben 100.0000 1000.0000 1000.0000 1000.0000 1000.0000
Manufacture of Liposomal Compositions:
Phases A and B are separately heated to 80 C.;
Phase A is stirred into Phase B (1-2 min (200 Upm))
Phase C is added at 60 C. (500-600 Upm)
Homogenisation at 50 C./2 Min 3000 Upm (U-Turax T-50)
Phase D is then added (stirred in) at <40 C.
optionally, pH is measured and adjusted to pH 6, if necessary (10% citric acid)
Skin Roughness (R.sub.Z)

(332) Evaluated are the changes in the parameter R.sub.Z in the test product treated areas in comparison to the changes in the area treated with the placebo (A) and to the changes in the untreated control area. The absolute changes by area and time point are shown below in figure one. A decrease in R.sub.Z corresponds to an increase in skin smoothness.

(333) FIG. 1 illustrates that the testproduct with 4% RM-0 significantly improves skin smoothness compared to the untreated control. This increase is further boosted by incorporating the Cyclopeptide Cyclo-(Arg-Gly-Asp-DPhe-Acha) (=CPPC 1) inside the liposomes (FIG. 01, testproduct 4% RM-100).

(334) Wrinkle Depth

(335) Evaluated is the parameter R.sub.Max in comparison to the initial condition in the respective test area and between the test product and placebo treated areas. The absolute changes by area and time point are shown in figure two. A decrease in R.sub.Max corresponds to a decrease in wrinkle depth.

(336) FIG. 2 illustrates that the wrinkle depth has been reduced after application of the placebo as well as the basic formulation containing 4% RM-1000 (containing 0.004% of the cyclopeptide Cyclo-(Arg-Gly-Asp-DPhe-Acha) (=CPPC 1). After 4 weeks treatment the depth is further decreased by RM-1000, containing 0.004% Cyclopeptide (=CPPC 1) compared to the placebo and significantly with respect to the beginning (FIG. 02).

(337) The reduction of the wrinkle volume has been visualized in FIG. 03.

(338) FIG. 3 illustrates the topography of skin surface. (A.) Colorization of wrinkle depth in mm (B.) Surface before treatment, t=0 (C.) Surface after 4 weeks treatment with 4% RM-1000 (containing 0.004% CPPC 1).

Example 18: Formulations/Dosageforms

(339) A: Vials

(340) A solution of 100 g of a cyclopeptide of the formula I and 5 g of disodium hydrogen phosphate in 3 I of double-distilled water is adjusted to a pH of 6.5 with 2 N hydrochloric acid, subjected to sterile filtration, dispensed into injection vials and lyophilized under sterile conditions, and the vials are sealed in a sterile manner. Each injection vial contains 5 mg of active principle.

(341) B: Suppositories

(342) A mixture of 20 g of a cyclopeptide of the formula I is melted with 100 g of soya lecithin and 1400 g of cocoa butter, and the mixture is poured into moulds and allowed to cool. Each suppository contains 20 mg of active principle.

(343) C: Solution

(344) A solution is prepared from 1 g of a cyclopeptide of the formula I, 9.38 g of NaH.sub.2PO.sub.42 H.sub.2O, 28.48 g of Na.sub.2HPO.sub.412 H.sub.2O and 0.1 g of benzalkonium chloride in 940 ml of double-distilled water. The pH is adjusted to 6.8, the solution is made up to 1 I and is sterilized by irradiation. This solution can be used in the form of e.g. eye drops.

(345) D: Ointment

(346) 500 mg of a cyclopeptide of the formula I are mixed with 99.5 g of petroleum jelly under aseptic conditions.

(347) E: Tablets

(348) A mixture of 100 g of a cyclopeptide of the formula I, 1 kg of lactose, 600 g of microcrystalline cellulose, 600 g of maize starch, 100 g of polyvinylpyrrolidone, 80 g of talc and 10 g of magnesium stearate is pressed to give tablets in a customary manner, such that each tablet contains 10 mg of active principle.

(349) F: Coated Tablets

(350) Tablets are pressed as stated in Example E and are then coated in a customary manner with a coating of sucrose, maize starch, talc, tragacanth and colourant.

(351) G: Capsules

(352) Hard gelatine capsules are filled in a customary manner with an a cyclopeptide of the formula I such that each capsule contains 5 mg of active principle.

(353) H: Spray

(354) 14 g of a cyclopeptide of the formula I are dissolved in 10 I of isotonic NaCl solution, and the solution is used to fill commercially available spray canisters having a pump mechanism. The solution can be sprayed into the mouth or nose. One spray burst (about 0.1 ml) corresponds to a dose of about 0.14 mg.

BRIEF DESCRIPTION OF DRAWINGS

(355) FIG. 1 illustrates that the testproduct with 4% RM-0 significantly improves skin smoothness compared to the untreated control, which increase is further boosted by incorporating the Cyclopeptide Cyclo-(Arg-Gly-Asp-DPhe-Acha) (=CPPC 1) inside the liposomes;

(356) FIG. 2 illustrates that the wrinkle depth has been reduced after application of the placebo as well as the basic formulation containing 4% RM-1000 (containing 0.004% of the cyclopeptide Cyclo-(Arg-Gly-Asp-DPhe-Acha) (=CPPC 1), and after 4 weeks treatment the depth is further decreased by RM-1000, containing 0.004% Cyclopeptide (=CPPC 1) compared to the placebo and significantly with respect to the beginning; and

(357) FIG. 3 illustrates the topography of skin surface, (A.) Colorization of wrinkle depth in mm (B.) Surface before treatment, t=0 (C.), and the surface after 4 weeks treatment with 4% RM-1000 (containing 0.004% CPPC 1).