COMPOSITION COMPRISING AN OXYGENATED CHOLESTEROL AND USE THEREOF FOR TOPICAL TREATMENT OF DISEASES

Abstract: A pharmaceutical composition comprising an oxygenated cholesterol and a penetration-enhancing agent which is useful for topical application to the skin of a patient suffering from a proliferative skin disease characterized by germinative cells having a rapid rate of replication, e.g. psoriasis. The compositon comprises an effective amount for the inhibition of germinative cell mitosis of an oxygenated cholesterol, e.g. 26-hydroxycholesterol, or a pharmaceutically effective derivative thereof e.g. an ester or ether. The invention is further directed to a method of treating a patient suffering from said skin disease comprising applying to the effected skin said therapeutic composition. The invention is also directed to the topical application of these compositions to the skin to decrease inflammation.

Description: COMPOS ITION COMPRISING AN OXYGENATED CHOLESTEROL AND USE THEREOF FOR TOPICAL TREATMENT OF DISEASES BAC_.GaθαND OF THE INVENTION 1. Field of the Invention This invention relates to pharmaceutical compositions for application to the skin and to a method for the treatment of prolif erative skin diseases. More particularly, this invention relates to topical compositions containing an oxygenated cholesterol, e.g. 26-hγdroxycholesterol, or a pharmaceutically acceptable derivative of an oxygenated cholesterol, e.g. an ester or an ether. This invention relates, particularly, to a topical composition containing 2β-hydroxycholesterol or cholest-l / 4-diene-2β-ol-3-one or a pharmaceutically acceptable derivative thereof and an agent for enhancing the penetration of these compounds into and through the skin. 2. Prior Art Prol if erative skin diseases are widespread throughout the world and afflict millions of humans and their domesticated animals. This invention prov ides a ethod for the treatment of such diseases and pharmaceutical compositions useful in allev-iating such diseases. As used herein, the expression "prolif erative skin diseases" means benign skin diseases which are characterized by epidermal cell proliferation, or division, and may also .be associated with incomplete tissue differentiation. Psoriasis is the most serious of the skin diseases with which this invention is concerned. Such diseases include: psoriasis, atopic dermatitis, non¬ specific dermatitis, primary irritant contact dermatitis, allergic contact dermatitis, lamellar ichthyosis, " epidermolytic hyperkeratosis, pre alignant sun induced keratosis, non-malignant keratosis, and seborrheic dermatitis in humans and atopic dermatitis and mange ' in domesticated animals. Heretofore, prolif erative skin diseases have been generally accepted by mankind as an ongoing evil having degrees of severity variable with time, with inherited skin traits and external factors, but always have been recognized as unsightly, painful, morbid diseases. Over the history of mankind, innumerable medicines and treatments have been proposed, tried and used with varying degrees of success. However, no treatment heretofore devised or pharmaceutical compositions used has been entirely successful in the wide spectrum of specific diseases encompassed by the expression prolif erative skin diseases. The present day treatments of commercial significance which are prescribed and used for the treatment of prolif erative skin diseases and associated inflammation include three approaches: (1) topical applications: coal tar derivatives, 5-fluorouracil, vita in A acid, glucocorticoids in high dosage, bath oils and non-specific emolient creams and ointments; (2) systemic administration: glucocor icoids and classic anti-cancer agents, for example, methothrexate, hydroxyurea, azaibine, cyclophosphamide; and (3) physical modalities: ultraviolet light, . irradiation, and in severe cases, surgery. Numerous compositions are known for the treatment of skin disorders and diseases by topical application which provide, in certain cases, some remission of the original symptoms, each treatment suffers some defect, for example, temporary and incomplete mitigation of symptoms, rapid re-occurrence of the disease when mitigation is terminated, serious and sometimes irreversible " damage (atrophy) resulting from the topical application for extended times of glucocorticoids, acute bone marrow suppression and cirrhosis of the liver resulting from the protracted use of methothrexate which may lead to death of the patient, and the causation of cancer by the application of anti-cancer drugs, irradiation, or ultraviolet rays. Psoriasis is perhaps the most prevalent type prolif erative skin disease. The early stage eruption of psoriasis may be non-specific in appearance and in such early stages is often confused with a ' variety of other skin diseases. The initial lesion is an erythematous papule which may progress to pustulation. Soon after the erythematous papules, the characteristic papulosquamous plaque appears. The* typical psoriatic eruption consists of erythematous, scaling plaques of variable size. The lesions are, in most cases, symmetrical. Clinically, the scale is distinctive, silvery and luxuriant in its pristine, untreated state. Underneath is a dull red surface which, upon removal of the scale, may show fine capillary bleeding points. Always sharply limited in border and frequently clearing in the center, psoriasis may come in any size and the scale m y in turn, range from being absent to extremely thick. . •, *," ^ ι Psoriasis, in addition. to bei;hg.-*an inflammatory disease, is a benign hyperplastic -disease of the skin. Epidermal cells in areas of skin involvement have a very rapid rate of replication. The mitotic index of the ' germinative cell population per unit length of involved epidermis is increased, and there is a reduced epidermal cell transmit time., ox epidermal cell "turnover" time in involved areas. Accordingly, the epidermis of the •psoriatic lesion grows very fast (about ten times normal rates) and sheds, large amounts of. scale. This is one of the key factors , in the pathology of the disease. In view of the foregoing, the principal thrust of the treatment protocol for psoriasis centers around use of antimetabolites or nicotinamide antagonists, such as the topical application of methotrexate, and folic acid antagonist; Azaribine, an orotidylate decarboxylase inhibitor (triacetyl-β-azauridine); hydroxyurea; 6-amino- nicotinamide; and systemically with mycophenolic acid. The first three drugs are antimetabolic agents and have been reported effective in producing remissions in patients with severe recalcitrant disease. However, all of these drugs have major side effects and can be given only in very severe cases and under extremely careful supervision by those experienced in their use. In many instances, some of the adverse effects of these drugs are worse than the psoriasis, particularly when using antineoplastic agents. In view of the foregoing, what is needed is a method for treating psoriasis by the topical application of a suitable compound to the psoriatic lesion. It would further be highly desirable to have a composition which is not only useful in the topical treatment of psoriasis, but other type prolif erative diseases, preferably by antimetabolites which characteristically inhibit mitosis. 26-Hydroxycholesterol, some of its derivatives and analogs, certain properties and some uses thereof are known, for example: Van iar ' efal. Biochemistry 6, 3269 (1967) and Ariπσer et al_ Biochem. Biophys. Acta., 665, 13 (1981) describes the finding that 2β-hydroxychoiesterol present, in human atheromata in vivo is of enzymatic origin. Kandutsch et al. Science 201, 498 (1978) reported that oxygenated sterols such as 25-hydroxycholesterol inhibit cholesterol synthesis in vitro, Javitt et al, "26-Eydrσxycholesterol Identification and Quantitation in Human Serum", J. Biol. Chem., 256:12644 (1981) describes the presence of 2 β -hydroxycholesterol (cholest-5-ene-3 beta, 26-diol) in biological fluids after neonatal life. Javitt et al. in J. Biol. Che ., 256:12644 (1981) describes the determination of 26-hydroxycholesterol in biological fluids by GLC-MS analysis using a deuterated analog. See, also, Biomedical Mass Spectrometry, 9:61-63 (1982) . U. S. Patent No. 4 ,427 ,668 to Javi tt (1984 ) describes a process for regulating cholesterol level in the body, particularly of tissue cholesterol, through the monitoring and/or administration of 26-hydroxycholesterol. Ad inistration is through the use of inj ectables, such as intravenous injectables, utilizing conventional pharmaceutical carriers for such use in tablet, capsul e, oral liquids or parenteral injectables. See also Japanese 58 172317 to Javitt et al which describes the use of 25-hydroxycholesterol compositions for depression of hyperplasia in smooth muscle cell. ' None of these foregoing references, however, teach or suggest the use of 26-hydroxycholesterol for the topical treatment of' skin disorders or diseases. Nor does any of these references disclose a method for passing 26-hydroxycholesterol through the skin directly into the blood stream without requiring an injection through the skin. It is therefore an obj ect of this invention to provide a new composition and method for treating prolif erative skin diseases. Another obj ect of the invention is to provide a method for treating prolif erative skin diseases by topical application of a therapeutic composition to the effected skin area. It is still another obj ect of this invention to provide novel pharmaceutically acceptable- compositions including an oxygenated cholesterol e.g. 26-hydroxycholesterol, or a pharmaceutically effective derivative thereof . It is yet another obj ect of this invention to provide cosmetic as well as therapeutic composi tions containing 26-hydroxycholesterol , or its derivatives which, when topically applied, w ill substantially alleviate the symptoms of various inflammatory skin disorders. It is yet another obj ect of this invention to provide compositions for transdermal delivery of an oxygenated cholesterol to a subj ect. SOM ARY OF THE INVENTION The present invention relates to the discovery that oxygenated cholesterols, e.g. 26-hydroxycholestεrol, or pharmaceutically acceptable derivatives thereof , when combined with a suitable carrier are surprisingly effective for treating prolif erative skin diseases such as psoriasis and inflammatory diseases. The invention is directed to a composition useful for topical application to the skin of a patient suffering f rom a prolif erative skin disease characterized by ger inative cells having a rapid rate of replication, said composition comprising an effective amount for the inhibition of germinative ceil mitosis of 25-hyάroxychol ester ol or a pharmaceutically effective derivative thereof and a pharmaceutically effective carrier therefor. The invention is further directed to a method of treating a patient suffering from a .prolif erative skin I disease characterized by germinative cells having a rapid rate of replication, comprising applying to the effected skin a therapeutic composition comprising a therapeutically effective amount for the inhibition of germinative cell mitosis of an oxygenated cholesterol, e.g. 26-hydroxycholesterol, or a pharmaceutically effective derivative thereof and a pharmaceutically effective carrier therefor. The invention is further directed to a therapeutic composition for topical application to --the skin of a patient suffering f rom an inflammatory skin disease characterized by. vasodilation and reddening of the skin, said composition comprising an effective amount for enhancing vasoconstriction and decreasing inflammation of the skin of an oxygenated cholesterol, e.g. 26-hydroxy cholesterol or a pharmaceutically effective derivative thereof and a pharmaceutically effective carrier therefor. Finally, this invention is directed to a method for carrying an oxygenated cholesterol, e.g. 2 S-hydroxy cholesterol, or a pharmaceutically effective derivative thererof, through skin or other body membranes which comprises combining said oxygenated cholesterol with a penetration-enhancing compound comprising a compound having the structural formula: Fherin R is Ξ or a lower alkyl group, .m is 3-7, n is 0-17 and R is -CH3, phenyl or substituted phenyl or to provide a composition including an effective amount of said oxygenated cholesterol and topically applying said compositions to the skin. DETAILED DESCRIPTION OF THE INVENTION As reported in THE MERCK INDEX, Tenth Edition, Published by MERCK δ CO., INC., Rah way, N.J. (1983) cholesterol has the followinσ structure: The active compound in the compositions and methods of this invention comprises an oxygenated derivative of cholesterol wherein the 26 position is substituted with a hydroxy, keto, or carboxylic acid group, i.e. the 26 position is oxidized. Other positions may also be oxidized. For example, the 3 and/or the 16 position may comprise a keto group. Preferably, the 26 position is substituted with a hydroxy group. Compounds which are within the scope of this invention include: 1) cholest-5-ene-3-ol-26-f ormyl-lβ-one 2) cholest-4-eπe-26-ol-3-one 3) cholest-l,4-diene-25-ol-3-one 4") 3 keto-cholest-l,4-dien-26-oic acid 5) cholest-5-ene-3 beta, 26-diol-lS-one (-25- hydroxycholesterol-lβ-one) 6) cholest-4-ene-26-ol-3 ,15-dione 7) cholest-l,4-diene-26-ol-3 , 16-dione 8) 3 ,16-diketo-cholest-l,4-dien-26-oic acid 9) 26-hydroxy cholesterol Th e stereo isomers of 25-hydroxychol esterol and the above-described oxygenated derivatives thereof, including the 25R and 25S stereo isomers and mixtures thereof, both naturally occurring and synthe tically produced, ar e also w ithin the scope of the present invention, in tha t they have an analogous inhibi ting effect on prolif erative diseases or inflammation. More pref erably, the active compound is selected from the - group consisting of 25-hydroxycholesterol and cholest-l,4-di eπe- 25-oi-3-one . The other compounds l isted above, may be derived from 25-hydroxycholesterol or cholesterol by oxidation. For exampl e, doubl e bonds may be introduced into the A- riπg and the 3-hydroxyl group oxidized to a ketone. Thus the A-ring of certain of the above compounds will have the f ormula : Formula 2 Formul a 3 1-4-diene These foregoing A- ring modif ied compounds may be produced by Oppenauer oxidation (see "Organic Reaction in Steroid Chemistry", Vol. 1 , Van Nostrand pp. 234-237 , Edited by Fried and Edwards (1972, incorporated herein by ref erence) of the silyl ether of 25-hydroxycholesterol , which is produced by standard silation reactions. This is subsequently treated with DDQ (dichlorodicyanobenzoquinone) ' to introduce double bonds at the 1 and 4 positions in the A-ring (see pages 308-318 of Fried and Edwards , above) . As used herein, as a compound .to be administered to a patient, the above oxygenated derivatives of . cholesterol include also "pharmaceutically acceptabl e derivatives" thereof. For exampl e, the mono and diesterif ied derivatives and mono and diether derivatives of any of the above compounds are included. Usually fatty acids, the same or analogous to those naturally occurring, are used to form the esters, but other inorganic and organic esters, such as the sulfates, carbonates and glucuronides, routinely employed in preparing pharmaceutically acceptabl e ester s can be used. Esterif ication and/or etherif ication can occur at the 3- and/or 26-position. Aryl and/or alkyl ethers, such as methyl, e thyl, or cyclo (e.g. cyclopentyl e thers ) are contemplated. Furthermore, acid salts can be used. Esterification and/or etherif ication can occur at the 3- and/or 26-position. Aryl and/or alkyl ethers, such as me thyl, ethyl, or cyclo (e.g. cyclopentyl ethers ) are contempl ated. Furthermore, acid sal ts and various substituted compounds,, f or exampl e, tho se containing el ements such as fluorine commonly used in modif ication of steroid-type compounds, as long as they are pharmaceutically acceptabl e, can be used in the method of the present invention. The therapeutic compositions of this invention are applied topically. The term "topical" as empl oyed herein relates to the use of the active compound incorporated in a suitable pharmaceutical carrier, and appl ied at the si te of the disease for exertion of local or transdermal action. Accordingly, such topical compositions incl ude those pharmaceutical forms in which the compound is applied externally by direct contact wi th the skin surface t.o b$ treated. Conventional pharmace tical f orms f or this purpose include ointments, lotions, pastes, j ellies, sprays, aerosol s, bath oils and the like. The term "ointment" embraces f ormulations (including creams) hav ing oil-soluble, water-solubl e or emulsion- type bases, e.g. pe trolatum, lanolin, polyethylene glycols, as well as mixtures thereof . Topical ointments can be prepared by dispersing the active compound or compounds in a suitabl e ointment base such as petrolatum, lanolin, polyethylene glycol, m ixtures thereof , and the like. Advantageously, the active compound or compounds is finely divided by means of a colloid mill util iz ing l ight l iquid pe trol atum as a lavigatiπg agent prior to dispersal in the ointment base. Topical creams and lotions are prepared dispersing the active compound or compounds in the oil phase pri or to the em ulsif ication of the oil phase in water. Oily components, emul sifier s, disper sing agents, gel atinizers and sol id material s which can be used to prepare such f ormulations are well known as those used in the preparation of cosmetics and topical products. Oily components, emulsifiers, dispersing agents and gelatinizers, of course, can be used alone or in combination with each other. It has been found that topical application with occlusion of an area larger than the medicated area produces improved results relative to nonoccl ' uded topical applications and is, therefore, the preferred method of topical treatment with the composition of this invention. In addition to conventional . formulations, the activity of the therapeutic compositions of this invention may be substantially 'enhanced by the use of skin penetrating vehicles, i.e., compounds which enhance percutaneous absorption of the active compound through the skin. Vehicles -which enhance the skin penetration of the active compounds disclosed herein include- dimethysulf oxide (DMSO), etc. As noted above, preferably the penetration- enhancing compound is selected from the group of compounds represented by the structural formula: wherein R' is H or a lower alkyl group, m is 3-7, n is 0- 17 and R is -CH 3 , phenyl or substituted phenyl or O II .C. — N : : ( H2)„ In a more preferred embodiment of the above penetration-enhancing compound when m is 3 and R is -CH 3 , then n is not 0-5. These compounds are described in the aforementioned U.S. Patent Nos. 4,424,210 and 4,316,893, the entire disclosures of which are incorporated herein by reference. The most preferred penetration-enhancing compound is AZONE, a t ' rademarked product sold by Nelson Research & Development Company, Irvine, California. This most preferred penetration-enhancing compound, l-dodecylazacycloheptan-2-one, is described in Stouσhtoπ, Arch. Der atol., Vol..118 (1982). The amount of the composition to be administered will obviously be an effective amount for the desired result expected therefrom. This, of course, will be ascertained by the ordinary skilled practitioner. In accordance with the usual prudent formulating practices, a dosage near the lower ends of the useful range of the particular agent may be employed initially and the dosage increased as indicated from the observed response, as in the routine procedure of the physician. The compositions of this invention comprise a pharmaceutical carrier and about .1% to 10% and preferably 1% to 51 by weight of the active compound relative to the total weight of the composition. If the amount of the active compound or its derivatives used is too low, the desired results of this invention will not be obtained or obtained insuffici ntly. The use of more of said active compound than required will not improve the desirable results in a manner proportional to the amount of compound used, and might bring about undesirable effects. For this reason, the above-mentioned range is preferred. Preferably, the active compound (which is generally difficult to dissolve in many otherwise suitable pharmaceutical carriers) is utilized with the aforementioned penetration-enhancing compounds, which is from .1% to 10% and preferably 1% to 5% by weight relative to the total weight of the topical composition of this invention. In carrying out the -novel method employing the topical composition, the active compound formulated, for example, as an ointment or solution, is applied to the affected area of the skin at a rate varying from about .1 mg/sq. cm of skin 'S-urf ace per day up to 10 mg./sq. cm of skin surface per day until the appearance of the affected, skin has returned to normal. The ointment or solution is generally applied for several days and in the case of- psoriasis, preferably using a continuous occlusive dressing. The concentration of active ingredients can vary from about .1% to about 10% by weight of the composition applied. With the foregoing concentration, a dose of about .01 ml/sq. cm of skin surface should supply an amount of active compound sufficient to achieve a desirable effect. The dosage of topical composition and length of time required for successfully treating each individual patient may vary, but those skilled in the art of medicine will be able to recognize these variations and adjust the course of therapy accordingly. One or more drugs selected from antibiotics, antihistiminics, anti ycotic agents and vitamins may be incorporated into the compositions of this invention to prepare a compounded preparation. In accordance with this invention, it has been found that prolif erative skin diseases and associated inflammation are alleviated, that is the symptoms of the disease are noticeabley improved or become undetectable, by the treatment of the afflicted patient, or animal, with one or more of the pharmaceutical compositions described in detail herein. A prolif erative skin disease can be considered to be effectively treated when there is a noticeable decrease in the thickness of a lesion to palpation, with or without residual redness, or residual slightly dilated blood vessels or residual hyper- or hypo-pigmentation. For example, psoriasis has been effectively treated when a scale-free psoriasis ' lesion is noticeably decreased in thickness, or noticeably but incompletely cleared or completely cleared. These effective treatments are all characterized by* an inhibition of mitosis of the germinative calls. By the use of the term "germinative cells" herein, it is meant the innermost layer of epidermal cells, which undergo mitosis. Additionally, the compositions of this invention may also be used to treat other skin disorders such as keratinizations and inflammatory diseases such as dry skin, ichthyosis, palmar and plantar hyperkeratosis, dandruff, acne, Darier's disease, pruritis, lichen simplex chroπicus, keratoses, warts, herpes, and eczema. Generally, this is accomplished by effecting vasomotor activity. Although the biochemical mechanism by which the active compound is effective for treating skin diseases and disorders is not fully understood, it is believed that the active compound may inhibit the activity of HMG Co A reductase, the rate limiting enzyme for the production of several compounds for cell growth, by reducing the amount of enzyme produced. This biological reaction would inhibit the mitosis of the generative cells. It has also been shown that 25-hydroxycholesterol is metabolized to bile acids by the liver. It is theorized that the extended side chain of these biologically active steroids, " in contrast to currently available glucocorticoids for the treatment of skin disorders, results in metabolism by -the liver. Thus, there is no interference with adrenal hormone synthesis and metabolism, reducing a major side effect of currently available glucocorticoids. The following examples identify certain compositions which typify the manner of combining selective active compounds with a pharmaceutical carrier for use in the process of treatment of prolif erative skin diseases as above generally described, but they are not intended to represent the limits of either the compositions or of the process of this invention which is defined in the claims. EXAMPLE 1 500 milligrams of 25-hydroxycholesterol is dissolved in 0.4 milliliter of AZONE with heating (80 °C) and then diluted with 9.6 milliliters of ethanol to produce 10 ml. of ethanol solution containing 4% of AZONE and 5% of 26-hydroxycholesterol. This method is used to prepare ethanol solutions comprising 5 percent 25-hydroxycholesterol and 4 percent AZONE and 1 percent 25-hydroxycholesterol and 1 percent AZONE. Solutions of 16-ketocholesterol, cholest-l,4-diene-26-ol-3-one are also prepared by this method. The solutions are evaluated for vasoconstricting activity as reported in EXAMPLE 2. EXAMPLE 2 Normal human volunteers 18-65 years of age are used in these studies. Compositions to be tested for vasoconstricting activity are -applied to the forearms so that four sites on each arm are treated with 10 mg or 10 microliters of the formulation. This is applied to " a 3 cm 2 area. The sites are varied randomly with regard to the formulations applied. The forearms are wrapped with occlusive film (Saran Wrap*) for 16 hours. The film is then removed and the arms washed with soap and water. Two hours after the removal of the Saran Wrap", the arms are read for vasoconstriction. The reader does not know which formulations were applied to which sites. The vasoconstrictor assay predicts the clinical effectiveness of pharmacological agents in psoriasis and eczematous diseases of the skin. It is a standard test used to develop agents for topical control of cutaneous inflammatory diseases including psoriasis. See, for example, the following references: 1. Cornell, R.C. and Stoughton, R.B.: Correlation of the vasoconstriction assay and clinical activity in psoriasis. Arch. Dermatol. 121.: 63-67, 1985. 2. Stoughton, .B. : Bioassay system for formulations of topically applied glucocorticosteroids. Arch. Dermatol. 1M: 825-827, 1972. 3. Barry, B.S., Woodford, R.: Activity and bioav ail ability of topical steroids: In vivo/ in vitro correlations for the vasoconstrictor test, J. Clin. Pharmacol.2:43-55, 1978. 4. Poulsen, B.J., Burdick, K., Bessler, S. : Paired comparison vasoconstrictor assays. Arch. Dermatol. l&lz 367-371 , 1974. 5. McKenzie, A.W. , Stoughton, R.B. : Method for comparing percutaneous absorption of steroids. Arch. Dermatol.- ___.: 608-610, 1962. Two separate studies were carried out with the results of the vasoconstrictor assay in each summarized in Table 1 and Table 2 below. - - Table 1 Vasoconstrictor Test (Anti-inflammatory Effect) Formulation Subjects' Scores T 16-ketocholesterol 1% in 1% Azone* in ethanol 0 0 0 0 0 0 0 0 0 0 16-ketocholesterol 5% in 5% Λzone* in ethanol 0 0 0 0 0 0 0 0 0 0 26-hydroxycholesterol 5% in 4% Λzone* in ethanol 0 1 0 0 1 1 0 0 1 0 26-hydroxycholesterol 1% in 1% Λzone* in ethanol 0 1 0 0 0 0 0 0 0 0 Ilydrocortisone 0.5% in ethanol 0 1 0 0 0 0 1 0 0 0 Ilydrocortisone 1.0% in ethanol 1 1 0 1 0 1 1 1 1 1 0 - no vasoconstriction 1 *- * ■ mild vasoconstriction 2 * - ** moderate vasoconstriction Note that 26-hydroxycholesterol was active in this assay, while 16-ketocholesterol w not have a hydroxy, keto or carboxylic acid group at the 26 position was inactive. Table 2 Vasoconstrictor Test (Λnti-Inflammatory Effect) Formulation Sub jects i 1 Scores To Cholest-l,4-diene-26-ol-3-one (1%) in 2 1 1 0 0 0 0 1 1 0 0.0% Λzone* in ethanol Cholest-l, 4-diene-26-ol-3-one (5%) in 1 0 1 1 1 0 0 0 0 0 4% Azone* in ethanol Hydrocortisone (1%) in ethanol 1 1 0 0 0 1 1 0 0 1 Ilydrocortisone' ( 0.2%) in ethanol 0 1 0 0 0 0 0 0 0 0 0 - No vasoconstriction 1 ~ Mild vasoconstriction 2 β Moderate vasoconstriction Note again an oxygenated cholesterol derivative having a hydroxy group in the 26 posi shows activity in the above assay. EXAMPLE 3 Example 2 is repeated, except the 1-n- dodecylazacyclo eptan-2-one is replaced w ith an equal amount of each of the f ollowing compounds : 1-n-hexy laz acy cl opentan-2- one l-n-heptyl azacyclopentan-2-one 1-n- octy laz acy cl ope n an-2- one l-n-nonylazacyclopentanτ2-one 1-n-de c laz acy cl opentan-2- one l-n-dodecyl azacycloρentan-2-one l-methylazac cl oheρtan-2-one 1-n-propyl azacy cl oheρtan-2-one l-n-butylazacycloheptan-2-one 1-n-pentyl azacy cl oheptan-2-one l-n-hexylazacycloheptan-2-one 1-n-heptyl azacy cloheptan-2-one l-n-octylazacycl oheptan-2-one 1-n-nonyl azacy cloheptan-2-one l-n-decylazacycl oheptan-2-one 1-n-butyl azacy clononaπ-2-one 1-n- octy laz acy clononan-2-one 1-phenyl azacy clopentan-2-one l-benzylazacyclopentan-2-one l- (2-chl orophenyl) azacy clopentan-2-one 1 ,3-Bis- ( 1-az acy cl opentan-2-onyl) propane 1 , 6-Bi s- (l-azacycloρentan-2-onyl) hexane Comparable results are obtained. EXAMPLE 4 Example 2 is repeated except that • 25-hydroxycholesterol is replaced with an equal amount of the following compounds: cholest-5-ene-3-ol-25-formyl-16-one cholest-4-ene-26-ol-3-one 3 keto-cholest-l,4-dien-26-oic acid cholest-5-ene-3 beta, 25-diol-16-one (25- hydroxycholesterol-15-one ' cholest-4-ene-26-ol-3 ,16-dione cholest-l,4-diene-26-ol-3, 16-dione 3 ,16-diketo-cholest-l,4-dien-26-oic acid Comparable results are obtained. It should be understood that various changes may be made in carrying out the above method and in the composition set forth above without departing from the spirit and scope of the invention. It is intended that all matter contained in the above description shall be interpreted as illustrative and not limiting.

Claims: We claim: 1. A method for treating an animal suffering from a prolif erative skin disease comprising topically administering to the skin of said animal an amount of an oxygenated cholesterol having a hydroxy, keto or carboxylic acid in the 26 position or a pharmaceutically acceptable derivative thereof effective to treat said prolif erative skin disease. 2. The method of claim 1 wherein said oxygenated cholesterol is selected from the group consisting of cholest-5-ene-3-ol-26-formyl-16-one cholest-4-ene-26-ol-3-one cholest-1 ,4-diene-25-ol-3-one 3 keto-cholest-l ,4-dien-26-oic acid cholest-5-ene-3 beta, 26-diol-lS-one (25- hydroxycholesterol- 16-one) cholest-4-ene-26-ol-3 ,16-dione cholest-l,4-diene-26-ol-3 , 16-dione 3 ,16-diketo-cholest-l ,4-dien-26-oic acid 25-hydroxycholesterol 3. The method of cl aim 2 wher ein said oxygenated cholesterol is 25-hydroxycholesterol. 4. The method of cl aim 2 wherein said oxygenated cholesterol is cholest-1 ,4-diene-26-ol-3-one. 5. The m ethod of cl aim 1 wherein sai d oxygenated cholesterol is administered in combination with a penetration- enhancing compound. -25- 6. The method of claim 5 wherein said penetration-enhancing compound is selected from the group of compounds represented by the structural formula: wherein R' is H or. a lower alkyl group, m is 3-7, n is 0- 17 and R is -CH3 , phenyl or substituted phenyl or o II .c. — ; : (CH * 2)„ 7. The method of claim 6 wherein when m is 3 and R is -CH3, then n is not 0-6. 8. The method of claim 7 wherein said penetration enhancing-compound is 1-n- dσdecylazacycloheptan-2-one. 9. The method of claim 8 wherein said oxygenated cholesterol is 26-hydroxycholesterol. 10. The method of claim 8 wherein said oxygenated cholesterol is cholest-l,4-diene-26-ol-3-oπe. 11. The method of claim 1 wherein said prolif erative skin disease is psoriasis. 12. The me thod of claim 9 wherein said di sease is psoriasis. 13. The method of claim 10 wherein said disease is psoriasis. 14. A method of treating a patient suffering from an inflammatory skin disease characterized by vasodilation and reddening of the skin, comprising applying to the effected skin a therapeutic composition comprising a therapeutically effective amount for enhancing vasoconstriction and decreasing inflammation of the skin of an oxygenated cholesterol having a hydroxy, keto or carboxylic acid group in the 26-position or a pharmaceutically effective derivative thereof and a pharmaceutically acceptable carrier therefor. 15. A topical composition comprising an oxygenated cholesterol having a hydroxy, keto or carboxylic acid substituent in the 26 position or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier for said oxygenated cholesterol. 16. The composi tion of claim 15 wherein said oxygenated cholesterol is selected from the group consisting of cholest-5-eπe-3-σl-2S-formyl-15-one cholest-4-ene-26-ol-3-one chol est-l ,4-diene-26-ol-3-one 3 keto-cholest-l / 4-dien-25-oic acid chol est-5-ene-3 be ta, 26-diol-15-one (25- hydroxychσlesterol-16-one ) cholest-4-eπe-25-ol-3,16-dione cholest-l,4-diene-26-ol-3, 16-dione 3,16-diketo-cholest-l,4-dien-26-oic acid 25-hydroxycholesterol 17. ' The composition of cl aim 15 wherein said oxygenated cholesterol is 26-hydrσxycholesterol. 18. The composition of claim 17 wherein said oxygenated cholesterol is cholest-l , 4-diene-26-ol-3-one. 19. The composition of cl aim 15 further comprising a penetration-enhancing compound. 20. The composition of claim 19 wherein said pene tration-enhancing compound is selected from the group of compounds represented by the structural formul a: wherein R' is H or a lower alkyl group, m is 3-7, n is 0- 17 and R is -CH3, phenyl or substituted phenyl or 21. The composition of claim 20 wherein when m is 3 and R is " -CI_3, then n is not 0-6. 22. The composition of claim 21 wherein said penetration enhancing-com pound is 1-n- dodecylazacycloheptan-2-oπe. 23. The composition of claim 22 wherein said oxygenated cholesterol is 25-hydroxycholesterol. 24. The composition of claim 22 wherein said oxygenated cholesterol is cholest-l,4-diene-26-ol-3-one.. 25. A method for traπsdermally delivering an oxygenated cholesterol having a hydroxy, keto or carboxylic acid in the 26 position to a subject which comprises; a) mixing said oxygenated cholesterol with a pene ration-enhancing amount of a compound represented by the following structural formula:. wherein R' is H or a lower alkyl group, is 3-7, n is 0- 17 and R is -CH3 , phenyl or substituted phenyl or and b) topically apply the mixture of step (a) to the subject. 25. The method of claim 25 wherein said oxygenated cholesterol is selected from the group consisting of cholest-5-ene-3-ol-26-formyl-16-one cholest-4-ene-26-ol-3-one cholest-l,4-diene-26-ol-3-one 3 keto-cholest-l,4-dien-26-oic acid cholest-5-ene-3 beta, 26-diol-16-one (26- hydroxycholesterol-16-one) cholest-4-ene-26-ol-3,15-dione ch*olest-l,4-diene-2S-ol-3, 16-dione 3,16-diketo-cholest-l,4-dien-26-oic acid 25-hydroxycholesterol 27. The method of cl aim 26 wher ein said oxygenated cholesterol i s 26-hydroxycholesterol. 28. The meth od of cl aim 26 wher ein said oxygenated cholesterol is cholest-l ,4-diene-26-ol-3-one. 29. The me thod of cl aim 26 wher ein w hen m is 3 and R is -CH3 , then n is not 0-6. 3 0. The method of cl aim 29 wher ein said penetration-enhancing compound' is 1-n- dodecylazacycloheptan-2-one . 31. The method of cl aim 3 0 w her ein sai d oxygenated cholesterol is 26-hydroxycholesterol. 32. The method of claim 30 wherein said oxygenated cholesterol is cholest-l,4-diene-26-ol-3-one. 33. The method of claim 14 wherein said . oxygenated cholesterol is selected from the group consisting of chσle__.t-jj-ene-3-ol-26-formyl-16-one cholest-4-ene-25-ol-3-one cholest-l,4-diene-26-ol-3-one 3 keto-cholest-l,4-dien-26-oiσ acid cholest-5-ene-3 beta, 25-diol-16-one (25- hydroxycholesterol-15-one) cholest-4-ene-26-ol-3 ,16-dione cholest-l,4-diene-26-ol-3 , 16-dione 3,15-diketo-cholest-l,4-dien-25-oic acid 25 hydroxy cholesterol 34. The method of claim 33 wherein said pharmaceutically acceptable carrier comprises a penetration-enhancing compound. 35. The method of claim 34 wherein said penetration-enhancing compound is selected from the group of compounds represented by the structural formula: wherein R' is H or a lower alkyl group, m is 3-7, n is 0- 17 and R is -CH 3 , phenyl or substituted phenyl or 36. The method of claim 35 wherein when m is 3 and R is -CH3, then n is not 0-6. 37. The method of claim 36 wherein said penetration enhancing-compound is 1-n- dodecylazacyclσheptan-2-one. 38. The method of claim 35 wherein said oxygenated cholesterol is 25-hydroxycholesterol. 39. The method of claim 35 wherein said oxygenated cholesterol is cholest-l,4-diene-26-ol-3-one.

Inventors: JAVITT NORMAN B, STOUGHTON RICHARD B, JAVITT, NORMAN, B., STOUGHTON, RICHARD, B.
Applicants: JAVITT NORMAN B, STOUGHTON RICHARD B, JAVITT, NORMAN, B., STOUGHTON, RICHARD, B.
Application year: 1986
Main classification: A61K31
IPC codes: A61K31, A61K47