MXPA97002272A

MXPA97002272A - Inhibition of cabe growth

Info

Publication number: MXPA97002272A
Application number: MXPA/A/1997/002272A
Authority: MX
Inventors: Shander Douglas; S Ahluwalia Gurpreet; Styczynski Peter
Original assignee: S Ahluwalia Gurpreet; Shander Douglas; Styczynski Peter
Priority date: 1994-09-28
Filing date: 1997-03-26
Publication date: 1997-06-01

Abstract

The present invention relates to the hair growth of a mammal is reduced by the application to the skin, of a composition that includes a protein kinase inhibitor.

Description

INHIBITION OF HAIR GROWTH DESCRIPTION OF THE INVENTION The invention relates to a method for reducing unwanted hair growth in mammals, and to a cosmetic method for this purpose. A main function of the hair of mammals is to provide protection against the environment. However, this function has been largely lost in humans, in which the hair is maintained or removed from various parts of the body essentially for cosmetic reasons. For example, it is generally preferred to have hair on the scalp but not on the face. Various procedures have been employed to eliminate unwanted hair, including shaving, electrolysis, depilatory creams or lotions, waxes, hair removal, and therapeutic antiandrogens. These conventional methods generally have drawbacks associated with them. Shaving, for example, can cause scratches and cuts, and may also promote the perception of an increase in the speed or proportion of hair regrowth. Shaving can leave a stubble. Electrolysis, on the other hand, can leave a treated area free of hair for prolonged periods of time, but can be expensive, painful, and sometimes leaves scars. Epilating creams, although very effective, are not typically recommended for frequent use due to their high irritation potential. The use of waxes and hair removal can cause pain, discomfort, and poor hair removal. Finally, antiandrogens - which have been used to treat female hirsutism - can have unwanted side effects. It has been previously described that the speed or proportion and the character of hair growth can be altered by the application to the skin of inhibitors of certain enzymes. These include the inhibitors of 5-alpha-reductase, ornithine-decarboxylase, S-adenosylmethionine decarboxylase, gamma-gluramyl-transpeptidase, and transglutaminase. See, for example, Breuer et al., US Patent No. 4,885,289; Shander, U.S. Patent No. 4,720,489; Ahluwalia, US Patent No. 5,095,007; Ahlu alia et al., U.S. Patent No. 5,096,911; Shander et al., US Patent No. 5,132,293; and Shander et al., US Patent No. , 143, 925. It has now been found that unwanted hair growth of the mammal (including man) - particularly androgen-stimulated hair growth - can be inhibited by the application to the skin of a composition that includes an inhibitor of protein kinase C ("PKC") in an amount effective to reduce hair growth. Unwanted hair growth, which is reduced, may be normal hair growth, or hair growth that results from an abnormal condition or illness. a PKC is a family of calcium-sensitive enzymes, dependent on phospholipids, which have the ability to phosphorylate proteins. PKC includes an ATP binding site, a calcium binding site, and a region that interacts with the phospholipid. Preferred inhibitors of PKC include those inhibitors that interact with one or more of these specific binding sites. Among the PKC inhibitors that can be used are 1) isoquinolin-sulfonamides such as 1- (5-isoquinolinyl-sulfonyl) -2-methylpiperizine and its derivatives (J. Biol. Chem. 264: 810-815, 1989); 2) bisindolyl aleimides such as 3- [1- (3-dimethylamino) propyl] -lH-indol-3-yl] -4- (lH-indol-3-yl) -lH-pyrrole-2, 5-monohydrochloride. -dione (GF109203X), Ro 31-7549, and derivatives of Ro 31-7549 (Biochem J. 294: 335-337, 1993; J. Biol. Chem. 266: 15771-15781, 1991; and J. Invest. Der atol 100: 240-246, 1993); 3) phenothiazi derivatives such as thioridazine, trifluoperizine, and triflucarbine (J. Dermat, Sci. 4: 18-25, 1992, and J. Biol. Chem. 255 8378-8380, 1980); 4) lyso-sphingolipids such as sphingosine and sphingosine derivatives (Science 235: 670-6''4, 1987, and Ann. Rev. Pharmacol., Toxicol., 32: 377-39", 1992); 5) staurosporine, and staurosporine derivatives such as 7-oxostaurosporine and 11-hydroxistaurosporine (Carcinogenesis 13: 355-359, 1992; J. Antibiot 45: 195-198, 1992; and J. Org. Chem. 57: 6327- 6329, 1992); 6) verapamil, phentolamine and imipramine (J. Biol. Chem. 255: 8378-8380, 1980); 7) 6-palmitate of L-ascorbic acid (Cancer Pos. ^: 6633-6638, 1987); 8) glycoside of tl i ri rr-t-ínic acid and 18β-glycyrrhetinic acid (Cancer < - > t * - »rs 49: 9-12, 1990); 9) polymyxin B, sangivamycin, and doxorubicin-Fe (III) (J. Dermat, Sci. 4: 18-25, 1992, J. Biol. Chem. 263: 1682-1692, 1988, and Trends in Pharmacol.Sci.12: 188-194, 1991); 1.0) the fungal product, balanol, isolated from Verti ci ll-Um bal anoi des (J. A. Chem. Soc. 115: 6452-6453, 1993); 11) substituted indolocarbazoles (Bioorg.

Med. Chem. Let. 3: 1959-1964, 1993); 12) 2- (aminomet: .l) piperidines (J. Med. Chem. 34: 2928-2931, 1991); 13) curcumin (FEBS Letters 341: 19-22, 1994); 14) 4-propyl-5- (4-pyridinyl) -2- (3H) -oxazolone (Cancer Research 52: 1195-1200, 1992); and 15) dekinin (Trends in Pharmacol.Sci.12: 188-194, 1991). Inhibitors can be irreversible or reversible (competitive and non-competitive). The PKC inhibitor is preferably incorporated into a topical composition that includes a dermatologically acceptable, non-toxic carrier or carrier, which is adapted to be spread on the skin. Examples of suitable carriers are acetone, alcohols, or a cream, lotion, or gel that can effectively distribute the active compound. A vehicle of this type is described in copending application PCT / US 93/0506A. In addition, a penetration enhancer can be added to the vehicle, in order to improve the effectiveness of the formulation as well. The concentration of the inhibitor in. The composition can be varied over a wide range, up to a saturated solution, preferably from 0.01 to 30% by weight or even more; The reduction "! Hair growth increases as the amount of inhibitor applied per unit area of the skin increases. The maximum amount applied effectively is limited only by the proportion at which the inhibitor penetrates the skin. In general, the effective amounts are in the range of 100 to 3000 micrograms or more per square centimeter of skin. The composition must be topically applied to a selected area of the body from which it is desired to inh: growth hair. For example, the composition can be applied to the face, particularly to the area of the facial beard, for example, the cheeks, the neck, the upper part of the lip and the chin. The composition can also be applied to the legs, arms, torso and armpits. The composition is particularly suitable for reducing unwanted hair growth in women suffering from hirsutism or other conditions. In humans, the composition should be applied once or twice a day, or even more frequently, for at least three months, to achieve a perceived reduction in hair growth. The reduction in hair firing is demonstrated when 1 frequency of hair removal (shaving, shaving with tweezers, use of hair removers, waxes) or reduced, or the person perceives less hair on or treated site, or quantitatively, when the hair is reduced. hair weight removed by shaving (for example, hair mass). The benefits of the reduced frequency of hair removal include convenience and less irritation to the skin. The intact Golden Syrian hamsters, males, are considered acceptable models for the growth of human beard hair, since they show organs of the sides of oval form, one on each side, each one of approximately 8 mm of greater diameter, which they grow as coarse and coarse black hair similar to the hair of the human beard. These organs produce hair in response to androgens in the hamster. To evaluate the effectiveness of a particular PKC inhibitor, the flanking organs of each of the hamster group are depilated by the application of a thioglycollate-based chemical epilator (Surgex). One organ of each animal applies 10-25 μl of vehicle only once a day, while the other organ of each animal is given an equal amount of vehicle containing a PKC inhibitor. After thirteen applications (one application per day for five days of a week), the flank organs are shaved and the amount of hair recovered (hair mass) of each is heavy. The percentage reduction in hair growth is calculated by subtracting the value of the hair mass (mg) from the side treated with the test compound, from the value of the hair mass from the side treated with vehicle; the delta value obtained is then divided by the value of the hair mass of the vehicle-treated side, and the resulting number is multiplied by 100. The test described above will be referred to herein as a "Golden Syrian hamster" test. Preferred compositions provide an inhibition in hair growth of at least about 35%, more preferably at least about 50% and even more preferably at least about 70%, when tested in the Golden Syrian hamster test. A number of PKC inhibitors were tested in the Golden Syrian hamster assay; the results are presented in Table I.

TABLE I hair mass (mean ± SEM) Compound Dose Vehículo3 H Untreated Treated% inhibition Verapamil 10% 5.5 1.45 ± .12 0.43 ± .05 69 ± 5 Thioridazine 10% 4.5 2.41 ± .19 0.74 ± .07 68 ± 4 Curcumin C 10% 5.5 1.66 ± 0.19 0.55 ± .ll ± 5 trifluoperizine 68 10% 7.5 2.38 ± .25 1.03 ± .16 56 ± 6 H-7 'A 1u% 6.0 1.76 ± .18 0.81 ± .08 54 ± 3 YOU 6-palmi ato L-Ascorbic acid 10% 8.5 2.82 ± .31 1.48 ± .14 46 ± 5 glycyrrhetinic acid glycoside 10% 4.0 1.841.19 0.94 ± .17 46 ± 10-glycyrrhetinic acid 18ff 7.5 B 75% 2.011.22 1.07 ± .10 44 ± 6 Imipramine A 10% 5.5 1.521.22 0.94 ± .18 38 ± 5 Fentola ina A 10% 6.0 2.05 ± .27 1.61 ± .22 19 ± 9 J Vehicle A includes pure water (68%), ethanol (16%), propylene glycol (5%), dipropylene glycol (5%), benzyl alcohol (4%) and propylene carbonate (2%); vehicle B includes ethanol (80%), pure water (10%) and dipropylene glycol (10%); and vehicle C includes acetone (40%), ethanol (20%), DMSO (20%), and water (20%). ? -7 is 1- (5-isoquinolinylsulfonyl) -2-methylpiperazine.

PKC activity was assessed in hair follicles isolated from the flank organs using a commercial assay kit obtained from GIBCO BPL (Gaithersburg, MD). The assay is based on the phosphorylation (incorporation of 3-P) of the acetylated myelin basic protein. After isolation, the hair follicles of flank organs were washed in phosphate buffered saline and homogenized in a buffer containing 20mM Tris, pH 7.5, 0.5 mM EDTA, 0.5% Triton X-100, ß- 10 mM mercaptoethanol, and 25 μg of each of the protease inhibitors, aprotinin and leupeptin. Homogenization of the hair follicle was added to the PKC reaction mixture at a final concentration of 10-20 μg / assay. The assay also included buffer, H0, phospholipid. The assay was performed in the presence or absence of «select PKC inhibitors. The volume of the reaction mixture was 50 μl, 32 P-ATP substrate was added in a volume of 10 μl. T, the reaction proceeded to 3 ß for 15 minutes, after which a 16.3 μl aliquot was removed from the reaction mixture, and r * »showed graphically on a paper filter. ! * 5 filters were washed twice in 1% phosphoric acid with gentle agitation for 5 minutes. The filters were then washed twice in H0 and placed in vials of scintillation. The incorporation of 32P, a measure of enzymatic activity, was determined using standard liquid scintillation techniques. significant inhibition with tisridazina (30% inhibition at 500 .mu.M) and trifluoperizine (42% inhibition at 500 uM) parts thereof which are thought to interfere with the binding site to fosfolípidos-- observed and H-7 , the most selective of PKC inhibitors and antagonist of the ATP binding site. There was an 86% inhibition of phosphorylation due to the inhibition of PKC activity, by 200 μM of H-7. The inhibition of PKC activity was almost 100%, with glycosyric acid glycoside (glycrylamine) at 200 μM; and 52% with 18β-glycyrrhetinic acid at 500 μM. It will be appreciated by those skilled in the art that the involution may be made within a wide range of equivalent parameters of the composition and conditions, without departing from the spirit or scope of the invention or any modality thereof.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention is that which is clear from the present description of the invention.

Having described the invention as above, property is claimed as contained in the following:

Claims

5. The process according to claim 1, characterized in that the inhibitor is trifluoperizine. 6. The process according to claim 1, characterized in that the inhibitor is 1- (5-isoquinolinylsulfonyl) -2-methylpiperazine. 7. The process according to claim 1, characterized in that the inhibitor is L-ascorbic acid 6-palmitate. 8. The process according to claim 1, characterized in that the inhibitor is glycoside glycoside. 9. The process according to claim 1, character! ? because the inhibitor is ldß-glycyrrhetinic acid. 10. The procuring agent according to claim 1, characterized in that the inhibitor is imipramine. 11. The process according to claim 1, characterized in that the inhibitor is phentolamine. 12. The process according to claim 1, characterized in that the inhibitor is an isoq inolin-sulfonamide. 13. The process according to claim 1, characterized in that the inhibitor is a bis-indolylmaleimide. 14. The process according to claim 1, characterized in that the inhibitor * »s a phenothiazine derivative. 15. The process according to claim 1, characterized in that the inhibitor < = > «* A lysophingolipid. 16. The procoso according to claim 1, characterized in that the inhibitor * »« a staurosporine. 17. The process according to claim 1, characterized in that the inhibitor -? select from the group consisting of polymyxin B, sangivamlcin, and doxorubicin-Fe (III). 18. The process according to claim 1, characterized in that the inhibitor is a balanol. 19. The process according to claim 1, characterized in that the inhibitor is a substituted indolcarbazole. 20. The process according to claim 1, characterized in that the inhibitor is a 2- (to inomethyl) piperidine. 21. The process according to claim 1, characterized in that the inhibitor is 4-propyl-5- (4-pyridinyl) -2- (3H) -oxazolone. 22. The process according to claim 1, characterized in that the inhibitor is dequalinium. 23. The process according to claim 1, characterized in that the inhibitor interacts with the ATP binding site in PKC. 24. The process according to claim 1, characterized in that the inhibitor interacts with the calcium binding site in PKC. 25. The process according to claim 1, characterized in that the inhibitor interacts with the PKC region that interacts with the phospholipid. 26. The process according to claim 1, characterized in that the inhibitor is an irreversible inhibitor. 27. The process according to claim 1, characterized in that the concentration of the inhibitor in the composition is between 1% and 30%. 28. The process according to claim 1, characterized in that the composition is applied to the skin in an amount from 100 to 3000 micrograms of inhibitor per square centimeter of skin. 29. The process according to claim 1, characterized in that the composition is applied to the skin on the face of said mammal. 30. The process according to claim 1, characterized in that the composition provides a reduction in hair growth of at least 30%, when treated in the Golden Syrian hamster test. 31. The process according to claim 1, characterized in that the composition provides a reduction in hair growth of at least 50%, when treated in the Golden Syrian hamster test. 32. The process according to claim 1, characterized in that the composition provides a reduction in hair growth of at least 70%, when treated in the Golden Syrian hamster test. 33. The process according to claim 1, characterized in that the mammal is a human. 34. The use of a protein kinase C inhibitor for the manufacture of a medicament for the inhibition of hair growth in mammals. 35. The use according to claim 34, characterized in that the inhibitor is as defined according to any of claims 2 to 26. 36. A method for the production of a composition for the inhibition of hair growth of a mammal, characterized in that it comprises the selection of a protein kinase C inhibitor, and the combination of said inhibitor, in an amount effective to reduce the growth of the hair, with a non-toxic vehicle or carrier, dermatologically acceptable. 37. A method according to claim 36, wherein the vehicle or carrier is adapted to be spread over the skin of a mammal. 38. A method according to claim 36, characterized in that the inhibitor is as defined according to any of claims 2 to 26. 39. The new use of a protein kinase C inhibitor for the reduction of hair growth. 40. A composition, when used for the inhibition of hair growth of a mammal, characterized in that it includes a protein kinase C inhibitor in an amount effective to reduce hair growth, and a non-toxic, dermatologically acceptable carrier or carrier. 41. A composition according to claim 40, characterized in that the inhibitor is as defined in accordance with any of claims 2 to 2 €.