USOO586 1142A United States Patent (19) 11 Patent Number: 5,861,142 Schick (45) Date of Patent: *Jan. 19, 1999

54 METHOD FOR PROMOTING HAIR, NAIL, 0 090 368 10/1983 European Pat. Off.. AND SKIN KERATINIZATION 0 187 012 7/1986 European Pat. Off.. 0 224 249 6/1987 European Pat. Off.. 76 Inventor: Mary Pichler Schick, 2027 Old Forge b 5. g i. As E. Way, Marietta, Ga. 30068 0 0 W.e "9CO * Notice: This patent issued on a continued pros- 95/16447 6/1995 WIPO. ecution application filed under 37 CFR OTHER PUBLICATIONS 1.53(d), and is subject to the twenty year patent term provisions of 35 U.S.C. Biochemistry Pharm., The interaction of Cat 154(a)(2). bamates with mammalian microtobule protein Vol. 28, p. 268O-2682. Davidse and Flach, “Differential Binding of Methyl 21 Appl. No.: 621,473 Benzimidazol-2-yl Carbamate to Fungal iustin S y 22 Filed: Mar. 25, 1996 Mechanism of Resistance to this Antimitotic Agent in 6 Mutant Strains of Aspergillus Nidulans”, Journal of Cell 51 Int. Cl...... A61K 7/06 Biology, vol. 72, 1977, pp. 174-193. 52 U.S. Cl...... 424/61; 424/701; 424/451; Fisher, et. al., “Efficacy of fenbenzalole and 424/464; 424/484; 424/489; 514/365; 514/395; against developing Stages of toxocara and toxascaris in 514/937; 514/944 dogs”, The Veterinary Record, vol. 132, No. 19, May 8, 58 Field of Search ...... 424/401, 701, 1943, pp. 473–475. 424/451, 464, 484, 489; 514/365, 395, 937, 944 (List continued on next page.) Primary Examiner Jvothsna Venkat 56) References Cited AOC, Agent, Or E". & Askew, LLP U.S. PATENT DOCUMENTS 57 ABSTRACT 35: 1919. E. s m r 5.83. A method for promoting keratinization of the hair, nails, and 3,954,791 5/1976 Loewe et al...... 260/309.2 skin on the body of an animal or human in which a 4,145,431 3/1979 Haugwitz et al. ... 424/273 B therapeutic amount of a benzimidazole Sufficient to cause 4,159,337 6/1979 Rowlands ...... 424/273 B keratinization is administered either Systemically or directly 4,166,858 9/1979 Rowlands ...... 424/273 B to the site on the body at which keratinization is desired. The 4,792,610 12/1988 Lachhein et al. ------548/329 method is useful for the treatment of wide variety of hair 4,871.544 10/1989 Eckenhoff ...... 424/438 loSS disorders in humans Such as alopecia, is useful for the 4,925,669 5/1990 Dyer et al...... 424/438 treatment of hair loSS disorders in animals, is useful for 5,169,8465,036,069 12/19927/1991 CrooksAndrews ...... et al. ...si,50s 514/249 enhancing the strength and length of fingernails and toenails 5340804 s/1994 wickiser. siso in humans, and is useful for enhancing the strength and 5,432,187 7/1995 Gazzard ...... 514/388 length of claws, horns, hooves and antlers in animals. The 5,434,163 7/1995 Edlind et al...... 514/310 method is also useful for the topical treatment of fungal infections, for Skin replacement or grafting, and for wound FOREIGN PATENT DOCUMENTS healing. 1 132906 10/1982 Canada. O O25 696 3/1981 European Pat. Off.. 5 Claims, 10 Drawing Sheets

3 4 5 time (weeks) 5,861,142 Page 2

OTHER PUBLICATIONS J.P. Seiler, “The Mutagenicity of Benzimidazole and Benz HammerSchlag, et. al., “Benomyl and imidazole Derivatives’, Mutation Research, Vol. 17, 1973, Methyl-2-benzimidazolecarbamate (MBC): Biomedical, pp. 21-25. Cytological and Chemical Aspects of Toxicity to Ustilago maydis and Saccharomyces cerevisae', Pesticide Biochem. J.P. Seiler, “Toxicology and Genetic Effects of Benzimida and Physiology, vol. 3, pp. 42-54. Zole Compounds”, Mutation Research, vol. 32 1975, pp. R.J. Horton, “ in a Wormy World”, Parasi 151-167. tology Today, vol. 6, No. 4, 1990 p. 106. Townsend and Wise, “The Synthesis and Chemistry Anthel D.W. Woolley, “Some Biological Effects Produced by Ben mintic Benzimidazoles”, Parasitology Today, vol. 6, No. 4, Zimidazole and Their Reversal by Purines”, (Laboratories of 1990, pp. 107-111. The Rockefeller Institute for Medical Research, New York) E. Lacey, “Mode of Action of Benzimidazoles”, Parasitol No. 17, 1943. ogy Today, vol. 6, No. 4, 1990, pp. 112-124. Jordan, et. al., “Endoparasitism in dogs: 21,583 cases B.L. Blagburn et. al., National Prevalence of Canine Para (1981-1990), IAVMA, vol. 203, No. 4, Aug. 1993, pp. sites based on Centrifugal Sucrose Flotation Examination of 547-549. Fecal Specimens: Preliminary Report (18); Proc. AAVP G.C. Coles, “The Biochemical Mode of Action of Some 1994, p. 57. Modern ”, Pestic. Sci. vol. 8, 1977, pp. C.K. Fenger et. al., “The Phylogenetic Relationship of 536-543. D. Diwel, “. II. Biological Properties and Sarcocystic Neurona to Other Coccidia Determined by Activity”, Pestic. Sci., vol. 8, 1977, pp. 550-555. Molecular Comparisons” (82), Proc. AAVP 1994, p. 57. H. Loewe, et. al. “Fenbendazole. I. Structure-Activity S.C. Barret. al. “Clinical Experience of Treating Cyrptospo Relationships", Pestic. Sci., vol. 8, 1977, pp. 544–549. ridiosis in Cats and Dogs with Paromomycin', (79) Proc. D.W. Gottschall, et. al., “The Metabolism of Benzimidazole Anthelmintics”, Parasitology Today, vol. 6, No. 4, 1990, pp. AAVP 1994, p. 56. 115-124. P.M. Schantz et. al., “Intestinal Parasites are Common in O.A. McKellar et al., “The Benzimidazole Pound Dogs in Fulton County, Georgia” (80); Proc. AAVP Agents-a review”, J. Pharmacol. Therap. Vol. 13, 1990, pp. 223-247. 1994 p. 56. M. Murray et al., “Hepatic Microsomal Metabolism of the B.T. Huss et al., “Fatal Cerebral Coenurosis in a Cat”, (70), Anthelmintic Benzimidazole Fenbendazole: Enhanced Inhi Proc. AAVP, 1994. bition of Cytochrome P450 Reactions by Oxidized Metabo lites of the Drug, Chem. Res. Toxicol. Vol. 5, 1992, pp. C.T. Faulkner et. al., “Gastrointestinal Parasitism in Dogs in 60-66. Tennessee (1986–1992)” (71), Proc. AAVP 1994. U.S. Patent Jan. 19, 1999 Sheet 1 of 10 5,861,142 SnstSo 9 i

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(uu) uplmous IIeu eulpnguo 5,861,142 1 2 METHOD FOR PROMOTING HAIR, NAIL, resemble Vellus hairs. In addition, as androgenic alopecia AND SKIN KERATINIZATION continues, the number of hairs in the active growth anagen phase decreases while there is an increase the number of The present method relates to the field of dermatology hairs in the telogen phase. and more specifically relates to a method for promoting the Telogen effluvium is a type of non-Scarring alopecia in which the anagen hairs prematurely move into the telogen keratinization of hair, nails, and Skin of an animal or human. phase. One most easily recognizable cause of telogen efflu BACKGROUND OF THE INVENTION Vium is the postpartum State in humans and other mammals. In the postpartum period, an increased number of hairs go For centuries, humans have been preoccupied with the into telogen due to the physical StreSS or hormonal changes length, thickness, color, and quantity of hair. Hair thinning, associated with delivery. Three to four months later, there is receding hairlines, and hair loSS can detrimentally affect considerable, but usually temporary hairloSS. The hair uSu appearance and Self-image and can even result in Significant ally returns to its normal state in six to twelve months. Other emotional consequences. causes of telogen effluvium in humans and other mammals Hair is a cylinder of keratinized cells protruding from a 15 include physical StreSS and Systemic illness, psychological hair follicle that anchors the hair in the skin. All hair follicles StreSS caused by major life events Such as a family death or in mammals have the same basic Structure. Hair follicles in divorce, medical nutritional deficiencies (kwashiorkor), or an adult human are generally arranged in groups of three. No absolute calorie deprivation (marasmus, crash diets), Vita new follicles are created or destroyed after birth, however, min and trace element deficiencies (Zinc, biotin, essential the type of hair produced by a given follicle can change. fatty acids, and iron), and endocrine abnormalities Most of the hairs on the Scalp are terminal hairs, which are (hypothyroidism, hyperadrenalcorticalism, hyperprolactine the coarse, pigmented, long hairs in which the bulb of the mia and adrenogenital Syndrome). hair follicle is seated deep in the dermis. The short, fine hairs Numerous remedies for hair loss have been attempted found on the Scalp and elsewhere on the body are Vellus ranging from wigs, toupees, and other hairpieces to the oral hairs. Vellus hairs are non-pigmented and unmedullated, 25 or topical administration of hair growth Solutions. No known having the hair bulb located Superficially in the dermis. cure has been discovered, even though many attempts have In humans, each hair follicle goes through repeated cycli been made. Hair transplantation is one method of treatment cal periods of growth including an active growth stage that has shown Some Success. Single hairs or plugs of thick, (anagen), which can persist for approximately 2 to 6 years; growing hair are transplanted from one region of the Scalp a transition phase (catagen), which lasts for only a week or to the site of hair loss. This method is very expensive, time two; and a resting period (telogen), which lasts 3 to 4 consuming and painful. Other hair growth Stimulation months. The hair is shed at the end of the telogen phase, and methods, including ultra-violet radiation, Scalp massage, a new hair is grown as the cycle repeats. In the human Scalp, revascularization Surgery and acupuncture, have been tried which contains approximately 100,000 hair follicles, nor with minimal or a total lack of Success. The pharmaceutical mally about 86% are in anagen, 1% are in catagen and 13% 35 drug minoxidil (2%, known commercially as Rogained, are in telogen. Therefore, in a normal human adult, approxi Upjohn Co., Kalamazoo, Mich.), originally discovered as a mately 100 hairs are shed from the scalp per day. vasodilator for the treatment of Severe hypertension, has Excessive hair loSS, or alopecia, may be classified as been found to be Somewhat useful as a hair growth Stimulant being one of two types, non-Scarring alopecia and Scarring in humans with androgenic alopecia. However, the drug has alopecia, and can be caused by a wide variety of factors. For 40 exhibited adverse effects. When taken orally, minoxidil has example, non-Scarring alopecia has been attributed to genet Serious cardiovascular side effects Such as fluid retention, ics and advanced age, administration of drugs. Such as tachycardia, and increased frequency of angina or new onset anti-cancer chemotherapeutic drugs and contraceptives, of angina, especially in perSons with poor coronary circu topical use of chemical treatments, Such as hair dyes, lation. Fluid retention can lead to weight gain, edema, heart permanent wave Solutions, and Straighteners, diseases, Such 45 failure and pleural or pericardial effusion. Although topical as leprosy or Syphilis, illness, allergy; and hair follicle administration of minoxidil has fewer Side effects, at least infection. Scarring alopecia may be a consequence of burns four months of continuous topical applications twice daily (accidental or post Surgical from cryoSurgery or laser are required before partial reversal of follicular miniaturiza Surgery) or trauma, which often causes follicle destruction. tion may be observed, resulting in larger, more pigmented Therefore, humans and other animals exhibiting Scarring 50 hairs and leSS hair Shedding, giving the appearance of hair alopecia may lack hair follicles in the region devoid of hair, growth. whereas those with non-Scarring alopecia possess hair fol Hair or fur loss is also prevalent in wild and domesticated licles with Short, fine, translucent hairs. animals, especially mammals. Although animals are not The most common type of human hair loSS is androgenic psychologically impacted by their loSS of fur or coat, their alopecia (also known as androgenetic alopecia), which is a 55 owners often are. In addition, fur loSS in animals whose fur non-Scarring hair loSS of telogen hairs caused by an exces is sheared or otherwise used commercially may cause fur Sive androgen effect in genetically Susceptible men and producers to Suffer economically. Animal fur or hair loSS has Women. Androgens trigger the miniaturization or atrophy of been attributed to many factors including disease, diet, terminal follicles that normally produce thick Scalp hair and metabolic disorders, insect bites, follicle infection, allergies, transforms them into Vellus-like follicles, eventually yield 60 and hot Spots or other hair or fur erosions caused by ing fine, downy hair that is barely perceptible. Androgenic excessive biting, chewing and Scratching. Currently, the alopecia is expressed in males as baldness of the vertex of only way to combat hair loSS in animals is to attempt to the Scalp and is commonly referred to as male pattern remove the underlying disorder or restrain the animal to baldness. In females, androgenic alopecia appears as diffuse prevent access to the hair loSS Site. Oftentimes the disorder hair loSS or thinning of the frontoparietal areas. AS alopecia 65 is Successfully treated, but the fur is not completely restored. progresses with age, hairs in these predisposed areas min Fingernails and toenails in humans and primates and their iaturize and appear to change from terminal hairs to corresponding claws, hooves, horns and antlers in animals 5,861,142 3 4 are composed of differently keratinizing cells. Human nails carbamates and benzimidazole prodrugs. BenZimidazole have lost most of their functional significance but remain prodrugs include phenylguanidines Such as, but not limited important for cosmetic reasons. Animals use their claws, to, fenbantel, netobimin and thiophanate. Preferred benz hooves, horns and antlers as weapons, tools and outward imidazole carbamates for use in the present method include, Signs of dominance. Nail disorders range from premature but are not limited to, thiabendazole, , breakage or roughness of the portion of the nail eXtending , , ciclobendazole, parbendazole, from the tip of the digit to total loss or destruction of the nail , fenbendazole, and metabolites, Such as fen plate. For example, nail breakage caused by the Splitting and bendazole sulfone, and derivatives thereof. The most pre resultant flaking of nails into layerS horizontal to the longi ferred benzimidazoles are fenbendazole, fenbendazole tudinal nail plate Surface, referred to as onychoSchizia, is prodrugs, and the Sulfoxide metabolites of fenbendazole, caused by abnormal keratinization. Such as (fenbendazole Sulfoxide). Nail disorders can also be caused by advanced age; The benzimidazole compound is combined with an appro infection from bacteria, fungi, yeast or mites (Scabies); priate carrier or Solvent to provide a benzimidazole compo trauma; congenital or hereditary defects, hyperplasias Such Sition. A preferred carrier is a pharmaceutically acceptable as warts, lesions, cysts and tumors, constant wetting of the 15 carrier. A most preferred carrier is a hair or nail cleansing, hand or foot, psoriasis, disease States Such as Darier's treatment, or cosmetic product Such as Shampoo or nail disease, Lichen planus, Alopecia areata, and twenty-nail polish remover. The benzimidazole compound is prepared in dystrophy, contact irritation or allergy to chemicals Such as granular, crystalline, powder, or amorphous form. The those contained in nail polish, hardenerS or adhesives, amorphous, Stabilized benzimidazoles and their metabolites metabolic disorders, Such as thyroid dysfunction; circulatory and derivatives are especially preferred because they are disorders, arthritis, Periungual telangiectasia, commonly more highly Soluble, bioavailable, and efficacious than crys observed in dermatomyositis patients and lupus erythema talline benzimidazole. The benzimidazole composition is toSuS patients, and pharmaceutical or illicit drug use. given Systemically, preferably by oral administration, or is Skin is the ultimate barrier against infection. Patients given locally by injection or topical application for Suffering from burns or other trauma to the Skin are at 25 parenteral absorption. The preferred method of administra extreme risk for disease caused by microorganisms. Skin tion is topical. grafting procedures have been used in Some circumstances, It is therefore an object of the present invention to provide however, limited Success has been achieved due to graft a Safe, inexpensive, and painleSS method for the treatment of rejection or insufficient skin growth to cover the injured hair loSS. area. A method for promoting in vitro autologous, epidermal It is a further object of the present invention to provide a cell propagation or enhancing in Vivo Skin growth or wound method for the prevention or minimization of hair loSS. healing would be extremely useful for replacing the skin It is a further object of the present invention to provide a barrier prior to the onset of life-threatening infection. method for Strengthening fingernails and toenails. The disadvantages of the hair loSS, nail claw, hoof, horn 35 It is a further object of the present invention to provide a and antler disorders, and skin replacement treatments cur method for Strengthening animal claws, horns, hooves and rently available are that they can be expensive, may cause antlers, especially the hooves of horses, particularly race adverse side-effects, and are not always effective for all horses. patients. Therefore, there is an on-going need for develop It is a further object of the present invention to provide a ment of new methods for treating hair or furloSS, nail, claw, 40 method for promoting normal hair and nail growth. hoof, horn or antler disorders, and skin replacement or It is a further object of the present invention to provide a wound healing in humans and animals. method for promoting skin growth and wound healing. SUMMARY OF THE INVENTION It is a further object of the present invention to provide a method for treating fungal infections of the skin and nails. A method for promoting keratinization on the body of an 45 It is a further object of the present invention to provide a animal or human is described. In accordance with the topical Scalp treatment composition to minimize hair loSS method, a therapeutic amount of a benzimidazole composi and a topical nail treatment composition to promote nail tion Sufficient to cause keratinization of the hair, nails or skin integrity. is administered. Administration may be either Systemically These and other objects of the present invention will or directly to the site on the body at which keratinization is 50 become apparent after reading the following detailed desired. description of the disclosed embodiments and the appended The method is useful for the treatment of a wide variety claims. of hair or fur loSS disorders in humans and animals caused by a wide variety of diseases and disorders, particularly BRIEF DESCRIPTION OF THE DRAWINGS androgenic alopecia and telogen effluvium. In addition, the 55 FIG. 1 is a graph showing follicle length of dorsal skin method is useful for normalizing keratinization in humans biopsies of hairless rats versus time in weeks after oral or and animals with abnormal keratinization processes that topical administration of fenbendazole. The “dot” symbol manifest themselves visibly as fingernails, toenails, claws or represents follicle length observed microscopically in a hooves that are Soft, easily bent and broken, frayed, split, or dorsal Skin biopsy of a hairleSS rat after oral administration deformed in shape. The method is also useful for enhancing 60 of fenbendazole. The “plus” symbol represents follicle the Strength and length of normal nails, for enhancing wound length observed microscopically in a dorsal skin biopsy of a healing or skin grafting by promoting the propagation of hairless rat after topical administration offenbendazole. The epidermal cells, and to combat cutaneous fungal infections, “asterisk” symbol represents follicle length observed micro particularly fungal infections of the nails and skin. Scopically in a dorsal skin biopsy of a negative control Abenzimidazole is a compound containing a bicyclic ring 65 hairless rat untreated with fenbendazole. Structure in which benzene is fused to the 4- and 5-positions FIG. 2 is a graph showing the number of vellus hairs of an imidazole and includes benzimidazoles, benzimidazole present on facial skin biopsies of hairleSS rats versus time in 5,861,142 S 6 days after oral administration of fenbendazole. The “dot' does not cause the production of new hair follicles, but symbol and the “asterisk” symbol represent the number of causes the growth of preexisting follicles wherein miniatur Vellus hairs observed microscopically in facial skin biopsies ized hairs become longer and thicker and may even develop of two hairless rats after oral administration offenbendazole. pigmentation. The “plus” symbol represents the number of vellus hairs The term “hair growth” as defined herein includes the observed microscopically in a facial skin biopsy of a nega promotion and maintenance of normal active (anagen) hair tive control hairless rat untreated with fenbendazole. growth following disruption of the hair growth cycle in any FIG. 3 is a graph showing nail plate distal thickneSS Stage Such as occurs when various chemotherapeutic drugs Versus time in weeks of each finger of the left hand of human are administered or caused by telogen effluvium, conversion Female 1 receiving topical applications of fenbendazole to of hairs from Vellus hairs to terminal hairs, an increase in the nail. The “dot” symbol is the thumb. The “plus” symbol hair Shaft length, an increase in hair shaft diameter, an is the index finger. The “asterisk” symbol is the digitus increase in hair shaft medullation, an increase in hair medius. The “square” symbol is the digitus anularis. The pigmentation, an increase in hair follicle length. The term “X” symbol is the digitus minimus. “hair growth” further includes improved hair growth, FIG. 4 is a graph showing nail plate distal thickneSS 15 enhanced hair growth, and restoration to normal hair growth. Versus time in weeks of each finger of the left hand of human The term “animal' as used herein refers to both human Female 2 receiving topical applications of fenbendazole to and non-human Species, particularly mammals, and the nail. The symbols are the same as in FIG. 3. includes, but is not limited to, domestic, productive, and FIG. 5 is a graph showing nail plate distal thickneSS breeding animals. Such as dogs, cats, cattle and other Versus time in weeks of each finger of the right hand of ruminants, horses, sheep, goats, pigs, camels, alpacas, human Female 1 receiving topical applications offenbenda llamas, water buffalo, donkeys, rabbits, fallow deer, reindeer Zole to the nail. The symbols are the same as in FIG. 3. and Similar mammals; pelt animals. Such as mink, foxes, chinchilla and raccoons, avian species or poultry Such as FIG. 6 is a graph showing nail plate distal thickneSS chickens, geese, turkeys, pigeons, ducks, and ostriches, and Versus time in weeks of each finger of the right hand of 25 human Female 2 receiving topical applications offenbenda animals Such as reptiles and amphibians. Zole to the nail. The symbols are the same as in FIG. 3. The terms "keratinization composition' and “keratiniza FIG. 7 is a graph showing longitudinal nail growth verSuS tion promoting composition' as used herein include human time in weeks of each finger of the left hand of human and Veterinary pharmaceutical, treatment, cleansing, and Female 1 receiving topical applications of fenbendazole to cosmetic compositions for administration to humans or the nail. The symbols are the same as in FIG. 3. animals. FIG. 8 is a graph showing longitudinal nail growth verSuS Although not wishing to be bound by the following, a time in weeks of each finger of the left hand of human proposed mechanism of action of benzimidazoles to pro Female 2 receiving topical applications of fenbendazole to mote keratinization involves a blockage or interference with 35 the effects of androgens, Such as testosterone and its the nail. The symbols are the same as in FIG. 3. metabolites, on the epidermal cells of the outer root sheath FIG. 9 is a graph showing longitudinal nail growth verSuS of hair follicles, possibly by interactions such as inhibition time in weeks of each finger of the right hand of human of cytochrome P-450 isozymes such as P-450IA1 in human Female 1 receiving topical applications of fenbendazole to keratinizing structures, and P-450IA1, P-450IIA1, and the nail. The symbols are the same as in FIG. 3. P-450IIB1 in rat keratinizing structures. It is believed that FIG. 10 is a graph showing longitudinal nail growth 40 the benzimidazole effectively opposes a genetic predisposi Versus time in weeks of each finger of the right hand of tion for premature miniaturization of hair follicles occurring human Female 2 receiving topical applications offenbenda during alopecia, particularly androgenic alopecia. Therefore, Zole to the nail. The symbols are the same as in FIG. 3. the method described herein includes the administration of compounds having the functional characteristics of DETAILED DESCRIPTION OF THE 45 benzimidazoles, namely cytochrome P-450 isozymes DISCLOSED EMBODIMENTS inhibitors, to inhibit the adverse effects of androgens on hair A method for promoting keratinization on the body of an growth, Such as alopecia, particularly androgenic alopecia. animal or human, particularly hair, nail, or skin The method is useful for the treatment of a wide variety keratinization, is provided. In accordance with the method, 50 of hair loSS disorders in animals. Such as alopecia, including a therapeutic amount of a benzimidazole composition Suf human androgenic alopecia of females and male pattern ficient to cause keratinization is administered either Systemi baldness, hair loSS due to telogen effluvium, hair loSS due to cally or directly to the site on the body at which keratini anti-cancer chemotherapy, oral or Subcutaneous contracep Zation is desired. tives and other drugs, hair loSS due to topical chemical Keratin is a Scleroprotein or albuminoid found predomi 55 treatment, Such as hair dyes, permanent wave Solutions, and nantly in epidermis cuticular Structures Such as hair, nails, Straighteners, and hair loSS due to disease, Such as leprosy or and horns. The term "keratinization' as used herein is Syphilis, illness, allergy; and hair follicle infection. The defined as keratin formation or growth of all keratinizing method is used either prior to anticipated hair loSS, Such as Structures and includes, but is not limited to, Skin before or during the administration of anti-cancer chemo re-epithelialization, especially after first or Second degree 60 therapeutic drugs or after an observation or diagnosis of hair bums of the Skin on the body of humans and animals, hair loss has been established. In addition, the method directly growth and regrowth from existing hair follicles, both in promotes faster, early onset of hair regrowth in humans or length and thickness, and the cornification or growth, both other mammals Suffering from disorderS Such as telogen longitudinally and in thickness, of fingernails, toenails, effluvium. claws, horns, hooves, and antlers. Keratinization, or the 65 The method is also useful for hair or fur loss disorders in promotion of keratinization, results in the production of animals caused by disease, diet, metabolic disorders, insect thicker, Stronger and longer keratin Structures. The method bites, infection of follicles, allergies, and hot spots or other 5,861,142 7 8 hair or fur erosions caused by excessive biting, chewing and which allows outgrowth of the infected keratinized structure Scratching. In addition, the method is useful for improving away from the body, thereby preventing or decreasing or enhancing the growth or thickness of normal hair or fur. contact of the fungal elements with the newly developing Furthermore, the method is useful for enhancing, improv keratinized Structures on which the fungus depends for ing or restoring the normal Strength, thickness, and length of 5 survival. Therefore, the promotion of keratinization contrib normal, abnormal, or diseased nails, including fingernails utes greatly to the fungistatic effect. and toenails in humans, and claws, horns, hooves and antlers BenZimidazole Compounds in animals. Nails, defined herein to include fingernails, The term “benzimidazole' as used herein is a chemical toenails, claws, horns, hooves and antlers, treated in accor compound containing a bicyclic ring structure in which dance with the present method are leSS Susceptible to benzene is fused to the 4- and 5-positions of an imidazole bending, cracking, peeling and breaking. Therefore, the and includes benzimidazoles, benzimidazole carbamates, method is useful for the treatment of onychoSchizia, also benzimidazole prodrugs, benzimidazole derivatives, and known as lamellar dystrophy, which is the horizontal Sepa benzimidazole metabolites. Phenylguanidines are Specifi ration of the corneocytes on the nail. The method is par cally included as Suitable benzimidazole prodrugs: ticularly useful for treating the hooves of horses, especially 15 Amorphous, crystalline and granular benzimidazole compo race horses, to enhance the Strength and thickness of the Sitions are included in the present definition of the term hoof to prevent or inhibit cracking, breakage, and fissuring, “benzimidazole'. Benzimidazoles having minimal toxic and in addition to prevent pain and infection. Side effects are preferred. Carbamates and, in particular, Nails treated in accordance with the present method are benzimidazole carbamates are used and can be used in large also less likely to show pitting of their Surfaces after dosages because they have exhibited a low incidence of Side treatment. Therefore, the method is useful for the treatment effects. of psoriasis of the nail bed, which causes nail pitting and deformation. The administration of a benzimidazole for the The preferred benzimidazole compounds are benzimida treatment of psoriasis of the nail bed may be in combination Zole anthelmintics. Anthelmintics are compounds useful for 25 treating helminthiasis in humans and animals. The term with other psoriasis treatments, Such as the administration of “benzimidazole anthelmintic' is defined herein as any methotrexate. In cases of Severe pSoriatic onychodystrophy, benzimidazole-containing agent known to act as a broad the remaining keratinous matter is first removed, Such as by Spectrum anthelmintic against endoparasites or nematodes applying 20% urea in a hydrolytic base. After the nail bed is Such as ascarids, hookworms, whipworms, roundworms and cleared, corticosteroids may be administered in combination kidneyworms. An analysis of the Structure-activity relation with the benzimidazole. ships of numerous benzimidazole compounds is provided in By promoting normal to increased keratinization, the the Scientific article of Loewe and Urbanietz, Fenbendazole. method described herein will allow a more economical I. Structure-Activity Relationships, Pestic. Sci. 8:544-549 production of keratin products Such as wool, hides, pelts and (1977), which is incorporated by reference herein. The horns. 35 anthelmintic mechanism of action of benzimidazoles com The method is also useful to improve or enhance the rate pounds appears to involve inhibition of the polymerization or extent of wound healing by promoting the propagation of of tubulin into microtubules within various helminth spe epidermal cells either in Vivo or in vitro for Subsequent cies. It is understood that one skilled in the art would be able transfer, or skin graft, to a patient Suffering from a disorder to Screen benzimidazoles for anthelmintic activity, and Such as trauma, burns, especially accident-induced burns, 40 thereby Screen for keratinization promoting activity, using abraded skin, ulcers, congenital malformations, or Surgery, Standard assays and techniques. in which large areas of skin must be generated. In addition, The preferred benzimidazole compound for use in the the method is useful for treating incised skin present from present method includes, but is not limited to, albendazole Surgical procedures including incision by Scalpel blades, (SmithKline Beecham Pharmaceuticals, Philadelphia, Pa.), Vaporization of skin by various types of laser emitted energy, 45 cambendazole (Merck/Univet, Rahway, N.J.), ciclobenda Such as caused by laser Surgery, necrotic skin Secondary to zole (Janssen/Cilag, Titusville, N.J.), fenbendazole cryoSurgical procedures or ionizing radiation induced burns (Hoechst-Roussel Agri-Vet, Sommerville, N.J.), flubenda caused by an external beam Such as cobalt radiation; or skin zole (Janssen, Titusville, N.J.), luxabendazole (Hoechst necrosis due implant radiation caused by cesium pellet Roussel Agri-Vet, Sommerville, N.J.), mebendazole implants. 50 (Janssen, Titusville, N.J.), oxfendazole (CooperS/Syntex, Techniques have been developed for Skin replacement in Palo Alto, Calif.), (Ciba-Geigy, Summit, which autologous epidermal cells are propagated in vitro N.J.), oxibendazole (SmithKline Beecham/Univet, and applied to the skin of the patient from whom the cells Philadelphia, Pa.), parbendazole (SmithKline Beecham/ were harvested. Such techniques are known to those skilled Ciba-Geigy, Philadelphia, Pa./Summit, N.J.), ricobendazole in the art. For example, epidermal cells can be impregnated 55 (Robert Young & Co., Wethersfield, Conn.), thiabendazole in a matrix, Such as a collagen matrix, to produce Skin of a (Merck, Rahway, N.J.), and metabolites, derivatives, and predetermined size or shape to be used for Skin grafting. prodrugs thereof, including the prodrugs febantel (CooperS/ The method described herein is additionally useful to Bayer, Shawnee Mission, Kan.), netobimin (Schering combat cutaneous fungal infections, particularly fungal Corporation, Kenilworth, N.J.), and thiophanate (Micro infections of the nails and Skin Such as dermatophytes and 60 Biologicals/RMB Animal Health). These benzimidazoles candidiasis by topical application of the benzimidazole are or have been commercially available from the companies composition to the nail or skin Surface. Fungal infections, listed above in parentheses. The most preferred benzimida including fungal infections of the nails, are particularly Zoles are fenbendazole and prodrugs thereof; the Sulfoxide prevalent in patients suffering from AIDS. The method metabolites of fenbendazole, Such as Oxfendazole, and the demonstrates antifungal effects not only because benzimi 65 (4! hydroxyphenyl)thio metabolites of fenbendazole. dazole compounds possess chemical fungicidal properties, Fenbendazole has been described in European Patent but also because benzimidazoles promote keratinization, Application Publication No. 090,368 to Ganley et al. for 5,861,142 10 administration to animals to treat helminthiasis. Ganley et al. thiophanate: 1,2-phenylenebis(iminocarbonothioyl) biscar State that fenbendazole is non-teratogenic and non bamic acid diethyl ester or 4,4'-O-phenylenebis3 carcinogenic and therefore safely used in animals at any thioallophanic aciddiethyl ester or 1,2-bis(3- Stage of pregnancy. In addition, Ganley et al. claim that the ethoxycarbonyl-2-thioureido)benzene drug has no adverse effect on fertility, and can be used at the BenZimidazoles useful in the method and composition time of conception in the female and the breeding male described herein include compounds having one or more of mammal and has no toxic or teratogenic effects on embryos the formulas set forth below as Formula 1, Formula 2 or or developing fetuses. Ganley et al. further describe fen Formula 3 and pharmaceutically acceptable Salts thereof. bendazole as having a very high Safety margin and is Preferred benzimidazoles for use in the compositions and non-toxic to the humans who are administering the drug to methods described herein have one or more of the formulas animals. According to Ganley et al., Oxfendazole and set forth as Formula 2. Most preferred benzimidazoles for albendazole have Substantially the same spectrum of activity use in the compositions and methods described herein have as fenbendazole, but a lower therapeutic index. In addition, one or more of the formulas set forth as Formula 3. AS in avian Species, fenbendazole fails to depress egg produc described in more detail below, the compounds are most tion or hatchability indices. preferably provided in an amorphous (non-crystalline) form A list of the foregoing benzimidazoles, including the 15 to enhance Solubility. phenylguanidine prodrugs, and one or more chemical names commonly associated there with, if available, are provided Formula 1: below: albendazole: methyl 5-(propylthio)-1H-benzimidazol-2-yl carb amate, 5-n-propylthio-2-carbomethoxy aminobenzimidazole, or methyl 5(6)-n-propylthio-2- benzimidazole carbamate cambendazole: isopropyl 2-(4-thiazolyl)-1H-benzimidazol 5-yl carbamate ciclo be n dazole (or cyclobe n dazole) : methyl 25 5-cyclopropylcarbonyl-1H-benzimidazol-2-yl carbamate O 5-cyclopropylcarbonyl-2- wherein Z is -H or halo; R is a five-membered het carbomethoxyaminobenzimidazole eroaromatic ring containing 1, 2, or 3 heteroatoms Selected fenben dazole: methyl 5-(phenylthio)-2- from O, N, and S, or R is -SCH, -NHCOOR, benzimidazolecarbamate or methyl 5-(phenylthio)-1H -NHCSOR, or -NHCOSR where R is alkyl, benzimidazol-2-yl)carbamate, methyl 5(6)-phenylthio cycloalkyl, alkenyl, alkynyl, phenyl, or naphthyl, and R is 1H-2-carbomethoxyaminobenzimidazole, or 5(6)- -H, R, or -XR, where X is O, S, SO, OS, -C(O), or phenylthio-2-benzimidazole carbamate -NHCOO-; and R is alkyl, cycloalkyl, aryl or aryl-alkyl, flub endazole: methyl 5-(4-fluorobenzoyl)-1H optionally Substituted with halo, alkyl, hydroxy, or alkoxy. benzimidazol-2-yl carbamate or 5-(4-fluorobenzoyl)-2- 35 carbomethoxyamino-benzimidazole Formula 2: mebendazole: methyl N-(5-benzoyl-2-benzimidazolyl) carbamate or methyl 5-benzoyl-1H-benzimidazol-2-yl N y carbamate or 5-benzoyl-2-carbomethoxy aminobenzimi R )- N dazole 40 N/ \COOR4 oxfendazole: methyl 5(6)-(phenylsulfinyl)-1H benzimidazol-2-yl carbamate or 5-phenylsulfinyl-1H-2- H carbomethoxyaminobenzimidazole oxibendazole: methyl 5-(1-propoxy)-1H-benzimidazol-2-yl wherein R is lower alkyl, and Rs is -S(O), R -OR, carbamate or 5-propoxy-2-carbomethoxy aminobenzimi 45 or -Y(CH),YR, where Y and Y are each indepen dazole dently O, S, or S(O), R is lower alkyl, phenyl, or naphthyl, parbendazole: methyl 5-(1-butyl)-1H-benzimidazole-2-yl and n is 1, 2, 3, or 4, R is lower alkyl, cycloalkyl, alkenyl carbamate or 5-(1-butyl)-2-carbomethoxy aminobenzimi of 3 to 7 carbon atoms, alkynyl of 3 to 7 carbon atoms, dazole phenyl, benzyl, phenylethyl, or naphthyl, and m is 0 or 1. thiabendazole: methyl 2-(4-thiazolyl)-1H-benzimidazole or 50 4-(2-benzimidazolyl) or 2-(4-thiazolyl)-1H Formula 3: benzimidazole R9 N triclabendazole: 6-chloro-5-(2,3-dichlorophenoxy)-2- X W methylthiobenzimidazole or 5-chloro-6-(2,3- C-NH-COOR8 dichlorophenoxy)-2-methylthio-1H-benzimidazole 55 R10 / febantel: 2-(methoxyacetyl)amino-4-(phenyl-thio) R11 N phenyl carbonimidoyl)biscarbamic acid dimethyl ester or dimethyl 2-(2-methoxyacetamido)-4-(phenylthio) wherein Rs is an alkyl of 1 to 4 carbon atoms, Ro and Rio phenyl-imidocarbonyldicarbamate are, independently, hydrogen, hydroxyl, alkoxy having 1 to netobimin: 2-(methoxycarbonyl)amino 2-nitro-5- 60 4 carbon atoms, halogen, trifluoromethyl, alkyl having 1 to (propylthio)phenyl)aminomethylene a mino 4 carbons atoms and carbalkoxy having 1-5 carbon atoms in ethaneSulfonic acid or 2-(methoxycarbonyl)amino 2 the alkoxy group, R is hydrogen or chlorine, fluorine, nitro-5-(propylthio) phenyl)iminomethylamino bromine, or iodine and X is oxygen, Sulfur, Sulfoxide or ethanesulfonic acid or methyl N'-(2-nitro-5-(propylthio) Sulfonate ester. phenyl-N-(2-sulfo ethyl)amidinocarb amate or 65 Combinations of compounds of Formulas 1, 2 and 3 are N-methoxycarbonyl-N'-(2-nitro-5-(propylthio)phenyl also useful. A preferred combination contains amorphous N"-2-(ethylsulfonic acid)guanidine compounds of Formula 1 or a combination of an amorphous 5,861,142 11 12 compound of Formula 1 and an amorphous compound of benzimidazole preparations are rarely Soluble in many phar Formula 2, especially where R is methyl. In a preferred maceutically acceptable carriers and are nearly insoluble in class of compounds of Formula 2, Rs is SR or S(O)R, and aqueous Solutions. Therefore, the formulation may be pre most preferably R is phenyl or n-propyl. Especially pre pared in an oil or oil-in-water emulsion. Suitable oils ferred combinations contain the compound fenbendazole, include, but are not limited to, arachis oil, peanut oil, olive most preferably in combination with Oxfendazole or tricla oil, Sesame oil, castor oil, corn oil; Synthetic triglycerides, bendazole. and Soluble polymers. The carrier or diluent may also The term “alkyl” as used herein is a saturated hydrocar include a delayed release material, Such as glyceryl bon radical containing 1 to 20 carbon atoms. The term monoStearate or glyceryl disterearate alone or in combina “lower alkyl as used herein is an alkyl radical of 1 to 6 tion with a wax. The composition may additionally contain carbon atoms. The term “cycloalkyl” as used herein is a conventional agents Such as preservatives (including anti cyclic Saturated hydrocarbon radical containing 3 to 8 car oxidizing agents Such as tocopherol), thickening agents, bon atoms. The term “alkenyl' as used herein is a hydro wetting and dispersing agents, buffers, humectants, Such as carbon radical of 3 to 7 carbon atoms, containing a double lactic acid and glycolic acid copolymers, emulsifying bond. The term “alkynyl' as used herein is a hydrocarbon 15 agents, fillers, emollients and Surface active agents (such as radical of 3 to 7 carbon atoms, containing a triple bond. The Sorbitane fatty acid esters). term “alkoxy” as used herein is a radical of the form RO-, Alternatively, the benzimidazole of a topical formulation where R is lower alkyl or cycloalkyl as described above. The is dissolved or Suspended in an aqueous Suspension con term “aryl” as used herein is an aromatic hydrocarbon taining benzyl alcohol, polyoxyethylene Sorbitan mono radical containing 6 carbon atoms, also known as a phenyl oleate, ethoxylated Sorbitane fatty acid esters, aprotic polar ring. The term “aryl-alkyl as used herein is an aryl group to Solvents, and, optionally, thickenerS Such as propylene which a lower alkyl group, as described above, is attached. glycol, polyethylene glycol, monohydric alcohol, n-methyl The term "halo' as used herein is a halogen radical Such as pyrrolidone, carboxymethyl cellulose, such as Methocel(R) fluoro, chloro, bromo, or iodo. A-15 Premium Carboxymethyl Cellulose (Dow Chemical Benzimidazoles to be used in the present method for 25 Co., Midland, Mich.) or other appropriate polymers. In promoting keratinization may be obtained commercially or addition, solvents such as dimethylsulfoxide (DMSO), Synthesized by conventional methods known in the art, Such DMSO glycol, decylmethylsulfoxide (decyl-MSO), dim as by methods set forth in U.S. Pat. Nos. 3,928,375 and ethylacetamide (DMA), dimethylformamide (DMF), satu 3,954,791, which are incorporated by reference herein and in rated fatty acids Such as lauric acid and alcohols, and weak the article by Townsend and Wise entitled “The Synthesis Surfactants containing a moderately sized polar group Such and Chemistry of Certain Anthelmintic Benzimidazoles”, as 1-dodecyl-azacyclohepten-2-one (AZone(E), E. L. Parasitology Today 6:107-112 (1990), which is incorpo Nelson), alcohols and ketones, particularly acetone, are rated by reference herein. useful for dissolving or partially dissolving the compound to Methods of Administration and Formulations enhance skin penetration of the benzimidazole. The penetra The benzimidazole compound, in granular, crystalline, 35 tion enhancement of benzimidazoles by Saturated fatty acids powder, or amorphous form, is provided in combination may be maximized when propylene glycol is used as the with a Suitable Solvent or carrier as a benzimidazole com vehicle. position for administration to a human or animal. The An additional topical benzimidazole formulation for benzimidazole composition is given Systemically by oral enhanced skin penetration employs the combined adminis administration or is given locally by injection or topical 40 tration of cyclodextrins, Such as carboxymethylethyl-3- application for parenteral absorption. Topical application cyclodextrin and a lipophilic penetration enhancer Such as includes transdermal or percutaneous delivery Such as by HPE-101 (1-2-decylthio)-ethylaza-cyclopentane-2-one) or application of a percutaneous occlusive patch. The preferred 1-dodecyl-azacyclohepten-2-one (AZone(E) in a topical method of administration is topical. vehicle. Topical administration should be directed to the site 45 For simplicity and ease of use for topical administration where keratinization is most likely to occur. The Site of to the body, the benzimidazole composition contains the keratinization in the Skin is the epidermis. The Site of benzimidazole compound in combination with a cosmetic, keratinization in hair or fur is the follicle. Therefore, for treatment, or cleansing hair or nail preparation Such as a hair topical treatment of allopecia, the benzimidazole composi Shampoo, hair rinse, hair conditioner, hair Spray, hair tion should be applied to the bald regions of the Scalp or 50 mousse, hair gel, nail lotion, nail ointment, nail paste, nail skin. Topical treatment of nail disorders should be applied at polish or nail polish remover. the cuticle base of the nail to maximize drug concentration Concentrated preparations containing a benzimidazole in in the vicinity of the keratogenous Zone, which lies beneath a concentration of 0.5 to 90 percent by weight, preferably 5 the lunula, or “white moon” shape at the base of the nail. The to 50 percent by weight, may be prepared and diluted before locations of these portions of the nail are well known to 55 administration. The preferred concentration to be adminis those skilled in the art and are described in dermatology tered topically is preferably from 5 to 50 mg benzimidazole texts such as on pages 557-559 of the dermatology text per kilogram body weight. Particularly preferred concentra HANDBOOK OF NON-INVASIVE METHODS AND THE tion employed are from 10 to 30 mg benzimidazole per SKIN, J. Serup and G. B. E. Jemac, eds., CRC Press, Boca kilogram body weight. A most preferred administration Raton, Fla., 1995. 60 concentration is 25 mg benzimidazole per kilogram body For topical administration, the benzimidazole, benzimi weight. dazole prodrug, benzimidazole metabolite or benzimidazole Poorly water-Soluble drugs. Such as benzimidazoles derivative is dissolved, Suspended or mixed as a paste, exhibit increased bioavailability when dispersed in a non cream or gel in an appropriate Solvent, homogenate, or aqueous mixture of a Surfactant and a co-Solvent in the carrier, Such as a pharmaceutically acceptable or cosmeti 65 presence of heat as described in U.S. Pat. No. 5,169,846 to cally acceptable carrier, to form a benzimidazole composi Michael J. Crooks, which is incorporated by reference tion. It is known by those skilled in the art that conventional herein. Formulations produced by the method of Crooks are 5,861,142 13 14 described as being free-flowing and completely miscible polysaccharides, including phase separation, Solvent with water and may be well Suited to topical administration. evaporation, emulsification, and Spray drying. Generally Amorphous benzimidazoles are preferred over crystalline polymers form the Supporting Structure of these or granular forms, particularly for topical administration, microSpheres, and the compound is incorporated into the due to their enhanced solubility in a variety of solvents. In polymer Structure. Exemplary polymers used for the forma addition, the amorphous benzimidazoles have a higher bio tion of microSpheres include homopolymers and copolymers availability and efficacy than crystalline benzimidazoles. of lactic acid and glycolic acid (PLGA). It will be under stood by those skilled in the art that microparticles, depend Amorphous benzimidazole derivatives are prepared as ing on their composition and size, may be administered described in European Patent Application Publication No. orally, by injection, or topically. 224,249 to Richard Allen Runkel, which is incorporated by An oral formulation is typically prepared by adding the reference herein. Basically, the amorphous compounds are additives to water and Stirring until dissolved, then adding produced by precipitating a benzimidazole derivative from the benzimidazole and Stirring until the mixture is homog an acidic or basic Solution by rapidly adding an amount of enous. The mixture is passed through a homogenizer, if base or acid sufficient to adjust the solution to the pH of necessary, to obtain relatively uniform particle Size distri lowest benzimidazole Solubility or by rapidly precipitating a 15 bution of approximately one micron. Homogenization is benzimidazole derivative from an acid or basic solution by achieved by means known to those skilled in the art, Such as adding a basic or acid Solution containing a Stabilizing with a rotor Stator or high pressure homogenizer. When amount of a Stabilizer polymer. Stable amorphous compo using a single head high pressure homogenizer, the mixture Sitions contain an amorphous benzimidazole compound in is passed through until the pressure can be maintained within combination with a Stabilizing amount of a Stabilizing the range of approximately 9,000 to 15,000 psig, preferably polymer Such as cellulose derivatives, polyvinyl pyrrollidone in a range of from about 12,000 to about 14,000 psig, most and derivatives, Xanthan gums, pectins, alginates, tragacanth preferably within a range of about 13,000 psig. When using and derivatives, gum arabic and derivatives, carrageenans, a triple head high pressure homogenizer, the mixture is agar and derivatives, polysaccharides from microbial passed through at a pressure of from about 2,000 to about Sources, arabinogalactans, galactomannans, and dextrans. 25 10,000 psig, preferably from about 4,000 to 8,000 psig. The most preferred Stabilizing polymer is a cellulose deriva The concentration of benzimidazole for oral administra tion is preferably between 0.5 and 50 mg per kilogram of tive such as methylcellulose, Methocel(R A-15, Methocel(R) body weight for a prolonged period of time. Generally, the A4C, or carboxymethylcellulose. The stabilized amorphous compound is administered initially at a high concentration benzimidazole compositions are appropriate for administra and the patient is then placed on a maintenance dosage tion topically or by parenteral injection In addition, the indefinitely. An alternative preferred concentration for oral Stabilized amorphous benzimidazole is useful in oral for administration is between approximately 2 and 20 percent mulations Such as Suspensions, tablets, or top dressing by weight for use on animals and between approximately 20 crumbles Specifically for mixing into animal feed or water as and 80 percent by weight for humans. described in more detail below. In avian Species, Such as chickens, pigeons, ducks and For oral administration, the benzimidazole compound 35 geese a 1% to 10% benzimidazole Suspension in drinking may be combined with an orally ingestible carrier formed water may be given for a Seven day period of time every two into tablets, capsules, powders, granules, pastes, to three weeks. A concentrated (about 10% to about 30%) homogenates, Solutions, Suspensions, emulsions, boli, medi benzimidazole formulation may be diluted with water to cated feed or added to drinking water. For delivery to yield a benzimidazole concentration from approximately animals, the benzimidazole composition is more easily 40 4,000 ppm to 10,000 ppm, preferably from about 6,000 ppm administered when combined with feed or drinking water. to 10,000 ppm, most preferably about 8,000 ppm. This Oral Solutions are prepared by dissolving, Suspending or diluted formulation may be used as a Stock Solution that is homogenizing the benzimidazole, benzimidazole prodrug, further diluted, for example, one ounce of the Stock formu benzimidazole metabolite or benzimidazole derivative in a lation is diluted in approximately a 1:128 ratio to obtain Suitable Solvent and, if appropriate, adding additives Such as 45 medicated drinking water having a benzimidazole concen Solubilizers, acids, bases, buffer Salts, antioxidants and pre tration of from about 45 to 80 ppm, preferably 65 ppm. Servatives. The Solutions may be filtered and packed under Alternatively, the concentrated benzimidazole formulation sterile conditions. Solvents that promote dissolution of the is diluted directly to a concentration of from approximately active compound in the main Solvent or Substances that 45 to approximately 80 ppm, preferably approximately 65 prevent precipitation of the benzimidazole may be used as 50 ppm and used for avian drinking water directly. The con solubilizers. Examples of such solubilizers include polyvi centration of benzimidazole is calculated to provide the nyl pyrrollidone, polyoxyethylated castor oil, and polyoxy targeted amount of benzimidazole per body weight of the ethylated Sorbitan esters. When the benzimidazole is pro avian or poultry being treated, preferably in the range of vided in an amorphous (non-crystalline) form, the preferred approximately 1 mg to 5 mg of benzimidazole per kilogram Stabilizer is a Stabilizer polymer as described in European 55 of body weight per day in the Volume of drinking water Patent Application Publication No. 224,249 to Runkel. normally consumed by the animal being treated in a 6 to 12 Exemplary preservatives include benzyl alcohol, hour treatment period, preferably an 8 hour treatment period. trichlorobutanol, parahydroxybenzoic acid esters and It will be understood by those skilled in the art that the n-butanol. particular dose or formulation for either topical or oral The benzimidazole may additionally be combined with 60 administration will be determined in part by consideration of conventional excipients and adjuvants Such as Starch, the animal or human undergoing treatment, the particular cellulose, talc, magnesium Stearate, Sugar, gelatin, calcium biological effects manifested by the benzimidazole and other carbonate, Silicic acid, carboxymethyl cellulose and the like. components utilized in the formulation, and the extended The compound may also be encapsulated within micropar period of time over which the effective treatment is desired. ticles or incorporated into a monolithic matrix, for Subse 65 The compositions and methods described above will be quent or Sustained release. Microparticles may be made further understood with reference to the following non using Synthetic polymers, natural polymers, proteins and limiting examples. 5,861,142 15 16 EXAMPLE 1. than the control for all Samples except the 28 day Sample, which was the same for both the oral treatment and untreated Oral and Topical Administration of Fenbendazole rat. The follicle length of the biopsy taken from the rat who to Hairless Rats to Promote Hair and Claw Growth had received fenbendazole topically was always less than An experiment was performed to compare the effects of that of the orally treated rat, but was higher than the control oral and topical administration of fenbendazole to hairleSS on day 14 and days 42-63. rats on hair and claw growth. Experimental Procedure: EXAMPLE 2 Circular dorsal skin biopsies (4 mm) were taken from adult CD(R Albino Hairless rats (CR1:CD(R) (SD)-hrBR, Oral Administration of Fenbendazole to Hairless Charles River Laboratories, Wilmington, Mass.) with a skin Rats to Promote Hair and Claw Growth punch prior to either oral or topical administration of fen An experiment was performed to determined the effects bendazole and at 7 day intervals thereafter for 35 days. on hair growth of an oral fenbendazole Suspension admin Dorsal Skin biopsies Samples were also removed from a istered to hairleSS rats. negative control rat who had received no fenbendazole 15 Experimental Procedure: either orally or topically. The biopsies were taken after Cylindrical dorsal skin punch biopsy Specimens (4 mm administration of isoflurane anesthesia and analyzed micro diameter) were taken from the dorsal facial area and the right Scopically by a dermatologic pathologist. lateral thigh area of adult CD(R Albino Hairless rats Topical Administration (CR1:CD(R) (SD)-hrBR, Charles River Laboratories, Finely ground fenbendazole powder (22%, Hoechst Wilmington, Mass.) at day 0 (prior to oral fenbendazole Roussel Agri-Vet, Sommerville, N.J.) was suspended to Suspension administration), day 17 and day 29 (after oral make a Supersaturated solution in 100% acetone. The dorsal fenbendazole Suspension or control Suspension body of an adult hairleSS rat was treated topically by administration). The skin punch biopsies were taken as applying the fenbendazole Suspension with a cotton Swab described above in Example 1. each day for 30 days. The head and tail were not treated. The 25 Thirty milliliters of a fenbendazole solution was prepared rat was not bathed, but was gently wiped with a damp cloth by mixing 1 teaspoonful of fenbendazole powder (22%, prior to each Subsequent treatment to rid the test Surface of Hoechst-Roussel Agri-Vet, Sommerville, N.J.) in 30 ml of granular debris. water to give a final concentration of 18 mg/ml. The Oral Administration fenbendazole Solution was placed in Separate water bottles The fenbendazole powder was diluted (one teaspoonful in for each of two rats receiving treatment. The rats consumed 30 ml water) to provide an 18 mg/ml aqueous Solution and the entire contents of the water bottles in a 24 hour period, was placed in a water bottle. The rat consumed the entire at which time a 30 ml aliquot of the fenbendazole solution contents of the water bottle in a 24 hour period, at which was resupplied for a 30 day dosing period. A negative time a 30 ml aliquot of the fenbendazole solution was control rat received an equivalent quantity of water. resupplied for a 30 day dosing period. 35 Results Results The rats receiving fenbendazole orally daily for 30 days The rat receiving fenbendazole orally exhibited hair exhibited hair growth on the face, lateral thorax, lateral growth on the face, lateral thorax and lateral abdomen by abdomen, and lateral thighs by day 7. The hair growth day 7. Hair growth on the face of this rat was patchy, Straight appeared white, patchy and Straight on all areas listed. The and white. Hair growth on the lateral thorax and lateral 40 muzzles exhibited long crimped to curling Vibrissae by day abdomen of this rat was white, fine and crimped. The hair 17. The claws on all four paws of the rats receiving fen growth on the lateral auxiliary area was Straight and white. bendazole showed significant growth in length, along with White hair was definitely visible on the rats treated orally increased periungual fold keratinization around each claw. and topically on the facial, Ventral neck, fore leg Hair growth and claw length and attendant keratinization circumferentially, hind leg circumferentially, tail region and 45 was minimal to absent on the negative control rat. paws by day 30 of treatment. The dorsum (where the skin Upon microscopic examination, no meaningful differ biopsies were taken) exhibited only a Small amount of hair ences were observed between treated and control animal growth. The muzzles of the treated rats contained long, dorsal skin biopsy Samples or follicle length. However, crimped Vibrissae. In addition, the claws on the treated measurable differences were observed between treated and animals showed a significant increase in length. Some hair 50 control animal facial biopsy Samples for the number of growth was observed on the control rat in the same general Vellus hair shafts. Agraph showing the number of Vellus hair areas as observed in the treated rats. However, the hair shafts from Samples taken from the facial areas in the treated growth on the control rat was minimal and Significantly leSS and untreated animals over the 30 day treatment period is Set than that observed on the treated rats. forth in FIG. 2. Upon microscopic examination, no measurable differ 55 ences were observed between treated and control animal EXAMPLE 3 dorsal skin biopsy Samples except for follicle length. A Topical Administration of Fenbendazole to graph of follicle length in the treated and untreated animals Fingernails to Increase Nail Plate Thickness and over the thirty day treatment period is set forth in FIG. 1. Length Follicle length as measured microscopically is the longest 60 distance from the granular layer of the epidermis to the base An experiment was performed to determined the effects of of the deepest follicle, measured vertically in millimeters. AS topical fenbendazole administration on fingernail plate shown in FIG. 1, both oral and topical fenbendazole admin thickness and length. istration resulted in an increase in follicle length that peaked Experimental Procedure: at day 14, Subsided and then increased gradually again from 65 Three teaspoonsful of finely ground fenbendazole powder day 35 onwards. Follicle length of the biopsy removed from (22%, Hoechst-Roussel Agri-Vet, Sommerville, N.J.) was the rat who had received fenbendazole orally was higher combined with 15 ml of 90% dimethylsulfoxide (DMSO) 5,861,142 17 18 and mixed well. The Solution was applied topically with a dropper and massaged gently into the uncut cuticular and TABLE 2-continued proximal nail fold Surfaces of dry, clean, fingernails of each finger of two human adult females. NAIL, PLATE DISTALTHCKNESS-RIGHT HAND Prior to treatment, the fingernails of the subjects exhibited Digitus Digitus Digitus a slow growth rate and Short length, primarily due to chronic Day Thumb Index medius anularis minimus breakage horizontally at the quick or distal portion of the 14 O.10/0.10 O.10/O.O5 O.10/O.O5 0.15/0.15 O.O7/O.05 nail. Nail breakage was associated with Soft, easily bent 21 O.30/O.30 O.2O/O.2O O.30/0.10 0.40/0.5O O.1O/O.2O nails or horizontal Splitting or flaking as the nails grew away 28 O.40/O.30 O.25/O.2O O.40/O.2O O.6O/O.7O O.25/O.30 from the nail bed. All nails were cut back to the nail bed to 35 0.40/O.30 O.30/0.30 0.40/O.30 0.60/O.80 O.30/0.30 allow for confluent nail growth and equality of growth among fingernails during the course of treatment. The first measurement was taken on day 0. The fenbendazole Solution TABLE 3 was applied from day 1 to day 14. Subsequent measurements were taken on days 7, 14, 21, 28, and 35. 15 LONGITUDINAL NAIL GROWTH-LEFT HAND All fingernails demonstrated relief from onychoSchizia Digitus Digitus Digitus during and after treatment. Nail thickneSS was maintained Day Thumb Index medius anularis minimus and progressively increased during the 35 trial period. All O 14.0/11.0 12...Of 11.0 12.0/10.9 12.0/9.4 10.0/9.0 nails failed to show Signs of fragility, Such as bending when 7 15.2/11.8 12.7/11.5 13.0/11.1 12.8/10.0 10.4/9.4 digital preSSure was applied to the fingernail, and failed to 14 16.9/14.2 13.8/12.4 14.1/11.6 14.4/11.2 16.5/10.0 exhibit pitting, nail layer Separation, or horizontal flaking as 21 18.8/15.8 14.5/13.5 15.6/11.9 17.6/13.0 * 10.0/11.5 the nails grew longitudinally away from the nail bed edge. 28 21.Of 18.1 15.Of 14.4 18. Of 12.5 19.Of 16.4 12.4f12.6 Nail plate thickness of all 10 fingernails was measured at 35 22.Of 19.2 16.6/15.O 13*f 13.2 20.0f2O.O 13. Of 14.0 7 day intervals with a stainleSS Steel caliper at the central *Nail broke aspect of the free edge of the nail plate as the nail plate 25 emerged over the nail bed. The caliper was positioned to grip the free edge of the fingernail using Standardized pressure. TABLE 4 Measurements were made in fractions of a millimeter. At the LONGITUDINAL NAL GROWTH-RIGHT HAND time of measurement, all nails were clean and dry without the presence of other nail treatment, polish, or cleanser. The Digitus Digitus Digitus caliper was positioned So that artifacts could not be pro Day Thumb Index medius anularis minimus duced by angling the caliper. O 15.0/9.1 12.Of 10.2 12.0/10.1 12.0/9.0 10.0/8.1 Longitudinal growth of each nail plate was measured with 7 15.1/9.5 12.4/10.5 13.0/10.8 12.8/9.3 10.5/8.4 a Straight ruler in Sub-millimeter increments, viewed by use 14 16.5/11.0 13.0/11.O 15.4f11.4 14.0/10.O. 11.4/9.0 35 21 18.2/13.4 14.2f11.8 16.4/12.2 17.0/11.1 12.2/10.1 of a magnifying loupe of 10x magnification. 28 22.Of 15.2 16.5/12.8 17.1/13.0 18.6/12.4 13. Of 12.2 The results for each fingernail of each experimental 35 23. Of 18.0 17.Of 14.0 17.5/14.0 18.8/14.4 14.2f14.O Subject, human female 1 and human female 2, are shown in Tables 1-4 below and presented in graphical form in FIGS. 3-10. All measurements are in millimeters. The results for Modifications and variations of the present compositions each Subject are separated by a Slash mark in the tables with 40 and methods for promoting keratinization will be obvious to Female 1 to the left of the slash mark. those skilled in the art from the foregoing detailed descrip tion. Such modifications and variations are intended to come TABLE 1. within the Scope of the appended claims. 45 I claim: NAIL, PLATE DISTALTHCKNESS-LEFT HAND 1. A method for promoting keratinization resulting in hair Digitus Digitus Digitus or nail growth comprising administering to a human or Day Thumb Index medius anularis minimus animal an effective amount of a benzimidazole anthelmintic O O.OO/O.OO O.OO/O.OO O.OO/O.OO O.OO/O.OO O.OO/O.OO Selected from the group consisting of thiabendazole, 7 O.10/0.10 O.OO/O.1O O.OO/O.10 O.10/O.OO O.OO/O.05 50 14 O.30/0.2O O.10/O.2O O.05/0.10 O.10/O.2O O.05/0.05 cambendazole, mebendazole, flubendazole, ciclobendazole, 21 O.45/O.30 O.40/O.30 O.10/0.25 O.30/0.40 * 10.10 fenbendazole, Oxfendazole, albendazole, and OXibendazole. 28 O.6O/O.30 O.45/0.5O O.25/O.35 0.5O/O.80 -0.20 2. The method of claim 1 wherein the nail is infected with 35 O.6O/O.35 0.45/0.5O O.30/O.35 0.5O/O.80 O.10/0.30 fungus and an effective amount of the benzimidazole is *Nail broke off horizontally at the quick-unavailable for measurement 55 administered topically to the fungally infected nail. 3. The method of claim 1 wherein the benzimidazole is TABLE 2 administered orally. 4. The method of claim 1 wherein the benzimidazole is NAIL, PLATE DISTALTHCKNESS-RIGHT HAND administered topically. 60 5. The method of claim 1 wherein the benzimidazole is Digitus Digitus Digitus Day Thumb Index medius anularis minimus Selected from the group consisting of fenbendazole and Oxfendazole. O O.OO/O.OO O.OO/O.OO O.OO/O.OO O.OO/O.OO O.OO/O.OO 7 O.10/0.10 O.16/O.O5 O.O7/O.O5 O.OO/O.1O O.06/0.05