NZ235686A - Use of mono-, di- or oligosaccharides in the treatment of alopecia - Google Patents

Description

New Zealand Paient Spedficaiion for Paient Number £35686 235 6 8 6 Priority Date{s): Ccvrtplola Sp«clffoflllon Filod: ..'■Uvjif, m Publication Data:..

P.O. Journal No: JiU$y No.: Date: new zealand patents act, 1953 complete specification COSMB2IC COWOSITIOK «»5S ts°CTl990 J!*eftveo i /We, bokh keditec a/s, a Danish Limited company of Strandve jen 122, dk-2900 Hellerup, Denmark hereby declare the invention lor which I / we pray that a patent may be granted to jne/us, and the method by which it is to be performed, to be particularly described in and by the following statement: - A. by page la) 235 6 8 6 1 c» COSMETIC COMPOSITION Alopecia (baldness or deficient hair growth) is a condition which leads Co more or less disabling or discomfort to Che individual suffering therefrom, ranging from minor cosmetic disadvantages to 5 severe psychological consequences.

Alopecia has a number of etiologies.

The most common form of alopecia is androgenetlc alopecia, more accurately described as common baldness. Androgenetlc alopecia occurs in chimpanzees, orang-outangs and other primates as well as in man.

As is implied by the name, androgenetlc alopecia is induced by androgenic stimulation of hair follicles predisposed by the interdependent influences of genetic factors and of ageing. Despite its name, it affect both the male and the female.

The initial stage in the condemned follicles is probably the accumu-15 lation of 5a-dihydrotestosterone, the tissue-active androgen which inhibits the metabolism of such follicles.

» There is a marked racial variation in the incidence of androgenetlc alopecia. The disease is most frequent and severe in Caucasolds.

The earliest histological change is the appearance of foci of degene-20 ration in the lower part of the connective-tissue sheath of the follicles, with perivascular basophilic change. The follicle gradually shrinks, leaving beneath it a strand of sclerosed and hyaline connective tissue. However, even in areas of scalp in which almost all follicles are shore and small, producing at best only tiny vellus hairs, 25 there remain a few qulescenc terminal follicles which can be stimulated into growth to give false hopes of a cure.

Androgenetlc alopecia is a very widespread condition, at least in its less severe forms. Thus, during adolescence, uniform recession of the frontal hair-line occurs in 96Z of males and abou* 80S of females. 328819BAX01/A26/OP/MJ/KPJ/19.04.1990 235686 2 As mencioned above, there are a number of other etiologies of alopecia, such as the administration of chemicals, such as drugs. As examples of these may bo mentioned anticoagulants such as large doses of heparin, heparinoids and coumarins, cytostatic agents, triparanol 5 and fluorobutyrophenone, excessive consumption of vitamin A, occupational exposure to sodium borate, potassium thiocyanate, large amounts of bismuth. Also, oral contraceptives have been suspected of giving rise to the disease, and the same applies to propranolol, metoprolol, levodopa, and ibuprofen.

Also, alopecia may have nutritional or metabolic origin, or it may be caused by disorders of the central nervous system. Furthermore, the significant association of the atrophic state with alopecia areata has been stressed relatively recently in some populations. Also, an association between autoimmunity and alopecia areata has long been 15 recognized. Alopecia including areata is also often seen in association with skin disease such as scalp eczema, psoriasis and other dermatosis and also in association with systemic disease such as LE.

Today, no completely satisfactory treatment or prophylaxis of alopecia exists. Although topical administration of minoxidil can induce 20 regrowth, it is not considered the definitive therapy.

Heparin, heparinoids and related glycosaminoglycans have been suggested as effective in stimulating hair growth (e.g. DE 3543221 Al, GB 936 916, CB 1 098 935, EP 35 919, EP 182 756, EP 277 428, EP 279 244, EP 295 092, EP 297 455).

In EP 295 092, it is described that hyaluronic acid fragments comprising from 7 to 50 monosaccharide units terminating either with a glucuronic acid unit and/or a N-acetyl glucosamine unit, or an unsaturated derivative of one or both of these terminal units are useful as hair stimulating agents when topically applied to the 30 scalp. Such compounds can be characterized by being "naturally" occurring mucopolysaccharide moieties.

It has now surprisingly been observed that after 3-4 weeks of two daily applications of an ointment containing 10X w/w of sucralfate, 328819DA.001/A26/OP/MJ/KPJ/I9.M.1990 2 3 5 6 8 6 3 there was appearance of heir In the otherwise bald lateral-frontal area# in a AO-year-old male who had a normal conmon mala-pactern baldness. At the beginning, a "plume" appeared In the area, and after a few days the "pluae" began turning into real hair* and 0 days 5 after, there vera dotena of real hairs Which were Indistinguishable from other hairs o£ the scalp, opart front being shorter and all coloured, in contrast to the existing hair, which was partially greyed.

This observation is nost remarkable in view of the fact that aucral-10 fate or sulfated Bono-, di- or oligosaccharides have apparently not been auggeated aa means for treating or preventing eloped*. Also, the sulfated mono-, di- or oligosaccharides do not belong to any chemical or therapeutic group of compounds previously used or suggested for the treatment oC alopecia, such as minoxidil, vitamin A, 15 steroida, in particular triamcinolone, pyrimldine carbamate, squaric acid, allergens and psoralens in PUVA and naturally occurring mucopolysaccharides .

Sucralfate and other disaccharlde polysulfate-aluminium compounds have been Indicated as effective in alleviating the symptoms of 20 anorectal disease when topicslly applied to hunan skin, and as effective in promoting the healing of wounds (WO 89/00047), and sulfated oligoseccharldes, particularly mono- and dleaecharidas such as sucrose octasulfate, have been SMntloned as wound healing agents (EP 230 023). WO 89/03643 and WO 09/03646 disclose a broad range of phar-25 macological effects of sucralfate and sucrose octasulfate such as anti-inflammatory, ant1-infective, anti-malignant, skin-protective and antl-wrinkle and other effects after topical or systemic administration. WO 89/07932 discloses <:he use of sucralfate and sucrose octasulfate In the treatment of {.ingivltls and parodontosis.

Baaed upon the obaervations made in the present invention and the inventors' knowledge of sulfated saccharides, it Is reasonable to contemplate that the hair growth stimulating effect will extend not only to sucralfate and the eodiuai salt of sucroee octasulfate, but also to other related sulfated mono-, dl- and oligosaccharides, which 3tUlfBAMl/KU/Or/M/KM/ttM.t990 2 3 5 6 8 6 4 are of « type not naturally occurring In glyoosaninoglycan »truc-tures, SUMMARY OF THE INVENTION In one aapect, Che invention providea a Method of treating and/or 5 preventing alopecia, the method comprising administering to a patient In need thereof a therapeutically or prophylactioally affective amount of a sulfated mono-, di- or oligosaccharide or a derivative, salt or complex thereof.

In another aspect, the invention relates to the use of a aulfated 10 mono-, di- or oligosaccharide for preparing a composition for use in the treatment and/or prevention of alopecia.

In a further aapect, the Invention relates to the cosmetic use of a sulfated aono-, dl- or oligosaccharide for combating or preventing hair loss and/or preserving the natural colour o£ the hair.

IS DETAILED DISCLOSURE OF THE INVENTION The sulfated saccharide used in accordance with the invention may to a sulfated monosaccharide, for instance sulfated xylose, fructose, glucose, ribose, arabinose, galaotose, rhamnoae, fucoae, sorbose, psicose, tagatose or gulose, a sulfated dlsaacharide such as sulfated 20 lucrois, lactose, maltose or oellobiose, or a aulfated oligosaccharide such as sulfated maltotriose, maltertreoae, or a sulfated raffi-nose, which is an oligosaccharide comprising a sulfated sucrose moiety together with a,galactose moiety, or a sulfated melesltose, which is a sulfated sucrose moiety together with a glucose moiety. In the 23 present context, the term "an oligosaccharide" is a saccharide consisting of 3-20 monosaccharide units in accordance with the generally accepted terminology.

The sulfated mono-, dl- or oligosaccharide Is preferably a polyaul-fated or peraulfaced saccharide, which meana that two or more, poa- 32MiraA.m/A»/Or/MJ/KI>,J/l»jM.UM 235 sibly all, sulfur-contalnlng moieties are present as substituents on hydroxy groups of the carbohydrate moiety.

In some cases, the sulfated mono*, di- or oligosaccharide may be complexed with or form a salt with a metal, e.g. an alkali or alka-5 line earth metal such as Na, K, Ca, Mg or Ba, or Al, Zn, Cu, Zr, Ti, Bi, Mn or Os, or with an organic base (e.g. an amino acid). The currently preferred salts are potassium and sodium salts, and the preferred complex is Che aluminium complex. Furthermore, the mono-, di- or oligosaccharide may be in the form of a suitable derivative, 10 e.g. a mono-, di- or polyester of aliphatic carboxylic acids such as formic, acetic, propionic, butanoic, myristic or stearic acid.

Preferably, the composition of the invention contains a persulfated disaccharide, for example sucrose octasulfate.

The preferred sulfated disaccharide can be represented by the follow-15 ing formula: wherein R is H, S03[Al2(OH)5], SO3H or an acyl residue of the above-mentioned carboxylic acids, the groups designated R being the same or 20 different, with the proviso that at least one R represents a sulfate group, or physiologically acceptable salts or complexes thereof.

The particular preferred compounds according to the invention are sucralfate and the potassium or sodium salt of sucrose octakis(hydro-25 gen sulfate).

Sucralfate may also be termed sucrose octakis(hydrogen sulfate) aluminium complex. Its CAS number is 54182-58-0. The commercial producc 235686 6 is a white powder which is practically insoluble in water and most organic solvents; it is soluble in acids and alkalis. In practice, there may be slight variations in the chemical composition, e.g., due to the fact that the sulfation may be slightly incomplete so that the 5 product may, e.g., contain a certain proportion of molecules which are not octasulfated (persulfated), but rather less sulfated such as heptasulfated. Such minor variations in the commercial product are well known and are reflected in the fact that e.g., the aluminium content in coircanrclal products may range from 17 to 21X and sulfur 10 from 9.5 to 12.5X. In the present context, the term "sucralfate" also comprises such generally accepted minor variations.

Sucralfate may, for instance, be prepared as disclosed in US 3,432,489 by reacting a 1-10X w/w aqueous solution of sucrose octasulfate or an alkali metal or alkaline earth metal salt thereof 15 with a 1-101 w/w aqueous solution containing aluminium ions, preferably A1C1(0H>2 at room temperature and a pH of 4-4.5. The sucrose octasulfate may be prepared by reacting sucrose with CISO3H, HjSO^ or H2SO4-C5H5N.

The sulfated saccharides may otherwise be prepared, for example, as 20 disclosed in EP 230023.

The sulfated sucrose is preferably selected from the group consisting of sucrose pentasulfate, sucrose hexasulfate, sucrose heptasulfate and sucrose octasulfate.

The administration form in which the sulfated saccharide is adrain!-25 stered will normally be a form suitable for topical application to the affected area. However, also administration into the skin or administration under the skin, e.g. by injection (by needle or der-majet) or other methods, are contemplated.

Although there may be cases where the sulfated saccharide may be 30 administered as such, it will typically be compounded with one or more pharmaceutical^ acceptable carriers or exclp.lents to present it in a form which is suitable for topical application or for injection or other form of introduction into or under the skin. In other words, 3288I9BA.Q01/A26/OP/MJ/KPJ/19.W.1990 23 5 6 8 6 it will be in Che form of a liquid, semi-solid or solid topical or systemic preparation such as an ointment, lotion, gel, cream, emulsion, solution, suspension, nicroemulsion, liposomes or as a roll-ball applicator, sponge applicator or a spray device; a shampoo, S hair tonic, hair conditioner, soap, balm, spray, paste, powder, sponge, strip, plaster, pad, dressing, or a comb or brush impregnated with or carrying the sulfated saccharide in such a manner that it is released when the comb or brush is used at the affected area.

The above-mentioned compositions are also suitable for cosmetic pur-10 poses where the compositions are applied to the hair or the skin areas normally covered with hair in order to prevent hair loss and/or to preserve che natural colour of the hair. Preferred compositions for cosmetic use are e.g. gels, emulsions, suspensions, liposomes, shampoos, hair tonics, hair conditioners, soaps, balms or sponges.

It may be interesting in certain cases Co combine che sulfated saccharide with other forms of therapy, in particular therapies known to either produce hair growth by themselves such as vitamin A, minoxidil, squaric acid, allergens, irritants, corticosteroids, or agents which attack or modify the mechanism responsible for the alopecia, 20 such as in the case of fungal infection, an ointment containing sucralfate and an antifungal agent such as ketoconazole, miconazole, clotrimazole, antiviral agents, such as acyclovir, antiinflammatory agents, antibacterial agents, etc. Also, ic may advantageous to combine the sulfated saccharide with ocher pharmaceutical products 25 known to have beneficial effects on the skin, such as vitamins, including vitamins fi, vitamin E, lactic acid, astringents, emollients, or ocher agents, such as hyaluronic acid or other glycosamino-glycans, dermatan sulfate, chondroicin sulfate, keratan sulfate, heparan or heparan sulfate. Normally, the sulfate saccharide will be 30 the predominant active component of the preparation.

Furthermore, advantages may be achieved by incorporating pharmaceuti-cally acceptable amounts of penetration enhancers in Che formulations, such as salicylic acid and other keratolytics, amino acids, thioglycollates, dinethylsulfoxide, and hydratlng agents, such as 35 glycerol. 328819BA.001/A26/OP/MJ/KPJ/19,04.1990 a 235 686 Plasters, spo.iges, strips, pads or other dressings may be prepared by Impregnating a dressing material such as cotton wool or gauze or a polymeric substance with a solution or suspension of the sulfated saccharide followed by drying. Alternatively, a paste, lotion, cream 5 or gel containing Che sulfated saccharide may be spread over the dressing material.

Alternatively, the sulfated mono*, di- or oligosaccharide may in certain cases be injected or otherwise introduced into or under the skin or scalp.

For topical application, the preparation may be formulated in accor-dance with conventional pharmaceutical practice wich pharmaceutical excipients conventionally used for topical, applications. The nature of the vehicle employed in the preparation of any particular composition will depend on the method intended for adminisCration of that 15 composition. Vehicles other Chan wacer that can be used in composi-cions can include solids or liquids such as emollients, solvents, huaectants, thickeners and powders. Examples of each of these types of vehicles, which can be used singly or as mixtures of one or more vehicles, are as follows: Emollients, such as stearyl alcohol, glyceryl monoricinoleace, glyceryl monosCearate, propane-1,2-diol, butane-1,3-diol, cetyl alcohol, isopropyl isoscearate, stearic acid, isobucyl palmitate, isocetyl •CearaCe, oleyl alcohol, isopropyl laurate, hexyl laurate, decyl oleace, octadecan-2-ol, isocetyl alcohol, cetyl palmitate, dimechyl-25 polysiloxane, dl-n-butyl aebacate, isopropyl myristate, isopropyl palmicace, Isopropyl scearace, butyl stearate, polyethylene glycol, triethylene glycol, lanolin, castor oil, acetylated lanolin alcohols, petroleum, mineral oil, butyl myriscace, isosCearic acid, palmicic acid, Isopropyl linoleace, lauryl lactate, myriscyl lactate, decyl 30 oleate, myristyl myristate; solvents, such as water, methylene chloride, isopropanol, castor oil, ethylene glycol monoethyl ether, diethylene glycol monobutyl ether, diethylene glycol nonoethyl ether, dimethyl sulfoxide, Cetrahydro- 328819BAJXH/A2S/OP/MJ/KW/19.04.1990 235686 9 furan, vegetable and animal oils, glycerol, ethanol, propanol, propylene glycol, and other glycols or alcohols, fixed oils; humectants, such as glycerin, sorbitol, sodium 2-pyrrolidone-5-car-boxylate, soluble collagen, dibutyl phthalate, gelatin; powders, such as chalk, talc, kaolin, starch and derivatives thereof, gums, colloidal silicon dioxide, sodium polyacrylate, chemically modified magnesium aluminium silicate, hydrated aluminium silicate, carboxyvinyl polymer, sodium carboxymethyl cellulose, ethylene glycol monostearate; gelling or swelling agents, such as pectin, gelatin and derivatives thereof, cellulose derivatives such as methyl cellulose, carboxymethyl cellulose or oxidised cellulose, guar gum, acacia gum, karaya gum, tragacanth gum, bentonite, agar, carbomer, bladderwrack, cerato-nia, dextran and derivatives thereof, ghatti gum, hectorite, ispaghu-15 la husk, xanthan gum; polymers, such as polylactic acid or polyglycolic acid polymers or copolymers thereof, paraffin, polyethylene, polyethylene oxide, polyethylene' glycol, polypropylene glycol, polyvinylpyrrolidone; surfactants, such as non-ionic surfactants, e.g. glycol and glycerol 20 esters, macrogol ethers and esters, sugar ethers and esters, such as sorbitan esters, ionic surfactants, such as amine soaps, metallic soaps, sulfated fatty alcohols, alkyl ether sulfates, sulfated oils, and ampholytlc surfactants and lecitins; 9 buffering agents, such as sodium, potassium, aluminium, magnesium or 25 calcium salts (such as the chloride, carbonate, bicarbonate, citrate, gluconate, lactate, acetate, gluceptate or tartrate).

The preparation of the invention may also contain other additives such as stabilizing agents, preservatives, etc.

Furthermore, it may be advantageous to provide modified release pre-30 parations in which the sulfated saccharide is incorporated into a 328819BA.001/A26/OP/MJ/KPJ/19.04.1990 235 6 polymer matrix, or nanoparticles, or liposomes or micelles, or adsorbed on ion exchange resins, or carried by a polymer.

The pharmacologically active element in sucralfate is probably the non-aluminium complexed sodiuia and/or potassium salt of sucrose occa-5 kis(hydrogen sulfate). Since such a salt is soluble in water, it would seem that a small particle size would be an important factor when preparing formulations of the sparingly soluble sucralfate. One way of achieving a small sucralfate particle size is by means of milling, grinding or disintegrating apparatus, e.g. a three roll mill, 10 where the sucralfate powder is ground, preferably together with a suitable liquid vehicle having a viscosity adapted tr effectively suspend the resulting fine particles, and preferably a relatively low vapour pressure so that no excessive evaporation with resulting agglomeration of the fine particles will occur, such as a polyalco-15 hoi, for example glycerin or polyethylene glycol, normally having a molecular weight in the range of 200-6000, such as PEG 400. The resulting preparation will normally contain up to 60-70X by weight of sucralfate particles with a fairly uniform particle size of about 5-10 pm or less (for 952 by weight of the sucralfate), the particles 20 being substantially evenly suspended in the vehicle. Such a paste can then be further suspended in any suitable pharmaceutical preparation using well known pharmaceutical methods. Another starting point for a small particle size sucralfate formulation is sucralfate "filter cake", which is an intermediary product obtained from the synthesis 25 of sucralfate. The "filter cake" comprises sucralfate with a content of water of about 50X by weight, and with a particle size o'f about 5-10 fim or less. This material can be mixed with, for instance, a water-miscible liquid which has a relatively low vapour pressure, such as glycerin, in order to prevent the water from evaporating, and 30 the sucralfate particles will retain their small size. Another important factor to Cake into consideration when preparing formulations of sucralfate and ocher sulfaCed saccharides is the strong negative charge of salts of sucrose octakis(hydrogen sulfate), and probably of most sulfated saccharides. The pharmacological effect of sucralface, 35 sales of sucrose ocCakis(hydrogen sulface) and other sulfated saccharides probably depends on this negatively charged entity, and che pharmacological effect of the drug may be reduced by the presence of 328819BA40I/A2</OP/MJ/KPJ/1?'(H1990 235 6 n posiCively charged mono- and divalent Ions In the vehicle. The person skilled In Che arc will be able Co Cake Chls lnco consideration, using guidelines from che relevanc literature, e.g., Harclndale, The Excra Pharmacopoeia, The Pharmaceutical Press, London, or ocher 5 pharmaceutical textbooks.

In Chis connection 1c should be mencioned that while the incorporation of sucralfate or ocher waCer-insoluble or sparingly water-soluble sulfated saccharides is besc performed as described herein Caking inCo consideration che physical and chemical properties of che 10 sulfated saccharide, in particular Che particle size consideraCions mencioned below, Che lncorporacion of wacer-soluble sulfated saccharides, such as sodium and potassium salts of sucrose oktakis (hydrogen sulfate) in preparations discussed herein will normally be extremely simple and will ordinarily consisC in che addition of che 15 sulfaced saccharide Co che preparaclon or to constituents thereof in either dry or dissolved form.

The sulfaced saccharide will norme'ily be used in a preparation in an amount of 0.001-992, typically abouC 0.1-752, such as about 0.2-302, preferably abouc 0.5-20Z, such as abouC 2-202, e.g., 3-152, by weight 20 of che Cocal preparaclon. For CherapeuCic and/or prophylaccic use, Che sulfaced saccharide may also be used in a preparaclon in an amounc of about 0.001-992 w/w, typically about 0.01-502 w/w, such as about 0.05-302 w/w, preferably about 0.1-202 w/w, such as abouc 0.5-152 w/w.

The concencracion of che sulfaced saccharide Co be used in each particular ease will of course depend upon che type of preparation and the intended use, but also on che solubilicy characterisCics of the sulfated saccharide and, for sparingly soluble and substantially insoluble sulfated saccharides, on Che particle size thereof; the sraal-30 ler Che parcicle size, Che faster will be che dissolution of even sparingly soluble or even substancially insoluble sulfated saccharides or complexes thereof. Insoluble or sparingly soluble salts or complexes of sulfated saccharides are preferably used in the form of a fine powder, for example having a particle size of 200 pm or less, 35 such as 100 pm or less. Examples of very small particle sizes which 3288!9DAA)1/A2</OP/MJ/K]PJ/19.04.1990 12 235 6 86 may be desirabls for certain purposes are e.g. 50 pm or less, such as 20 i*m or less, in certain cases 10 /im or less, such as 5 /jm or less.

A topical preparation containing the sulfated saccharide is normally administered between 1 and 10 times a day, depending on the formula-5 tion, the severity of the condition to be treated, the age of the patient, and other factors. Based upon experience with other substances used in the treatment of alopecia, in particular minoxidil, it is expected that the effect will depend on continuing application of the preparation, for several months, or even years. In view of the fact 10 that sucralfate is remarkably free of side effects, there should be no adverse long term consequences of such a treatment.

•» ' The invention is further illustrated by the following non-limiting examples.

EXAMPLE 1 Preparaclon of sodium snd potassium sucrose octasulfate I. Sucrose octasulfate 254.7 g (1.6 mol) of sulfur trioxide pyridine were slurried in 1300 ml of water-free pyridine. With stirring, 68.5 g (0.2 mol) of sucrose were added. The reaction mixture was heated to 65*C and kept 20 at this temperature for 240 minutes. As the reaction proceeded, the substance was separated as a thick flowing oil. When the reaction was terminated, the agitator was stopped, the pyridine phase was decanted, and the oily phase was dissolved in 600 ml of ion-exchanged water.

II. Potassium sucrose octasulfate One portion of sucrose octasulfate solution prepared as described in I above was adjusted with 10% w/w aqueous potassium hydroxide to pH -9 with stirring at room temperature. The solution was evaporated at 50'C in vacuo to remove pyridine and water until 880 g were left. The 32Sai9BA.001/A»/Or/MJ/KPi/i9XM.l?90 235 686 13 warn solution was filtered, adjusted to pH - 9.5, and the substance was precipitated with slow cooling to 5*C. The substance was filtered and washed with 300 ml of 1:1 ion-exchanged water/methanol and 300 ml of methanol. The wet filter cake was dried in vacuo at 50*C. The 5 crude product was dissolved at 40*C in 700 ml of lon-exchanged water.

The liquid was filtered and adjusted to pH - 9.5, and the substance was precipitated with slow cooling to 5*C. The precipitated substance was filtered and washed with 300 ml of 1:1 ion-exchanged water/methanol and 300 nl of methanol. The wee filter cake was dried in vacuo at 10 50*C. The substance was reprecipitated twice as mentioned above.

Yield: 137 g (about 532) of potassium sucrose octasulfate.

III. Sodium sucrose octasulfate One portion of sucrose octasulfate solution prepared as described In I above was adjusted with 10X w/w aqueous sodium hydroxide to pH - 9 15 with stirring at room temperature. The solution was evaporated at 50*C in vacuo to remove pyridine and water. When 250 ml were distilled, 580 ml of ethylene glycol were added and the evaporation was continued until 10 nor. vacuum. 100 ml of ethanol were added to the solution. The solution was filtered, and pH was adjusted to 9.5. 350 nl of ethanol were slowly added with vigorous stirring at 30- *C, and the substance will then precipitate. After the addition was ended, the mixture was cooled to 10'C, and the solid substance was filtered and washed with 50 ml of 1:1 ethanol/ethylene glycol and 200 ml of methanol. The wet filter cake was dried in vacuo at 50*C. 25 The crude product was dissolved in 520 ml of ethylene glycol and heated to 40*C, and 100 ml of ethanol were added. The solution was filtered and pH was adjusted to 9.5. 350 ml of ethanol were slowly added with vigorous stirring at 30-35'C and the substance will then precipitate. After the addition was ended, the mixture was cooled to 30 10*C and the solid substance was filtered. It was washed with 50 ml of 1:1 ethanol/ethylene glycol and 200 nl of ethanol. The wet filter cake was dried in vacuo at 50*C. The substance was reprecipitated once as above and then dried for 2 hours at room temperature in 500 ml of ethanol. The substance was filtered, washed with 200 ml of 35 ethanol and dried at 50*C in vacuo. 3288l9BA^l/A26/OP/MJ/KJ\J/19.04.t990 235 6 86 14 Yield: 146 g (about 63Z) of sodium sucrose octasulfate.

EXAMPLE 2 A creao consisting of che following ingredients is prepared (all percentages are by weight): 2:1 suspension of sucralfate* in polyethylene 15.OX glycol 400 (10X sucralfate in the final product) Lanolin 10. OX Vegetable oil (evenlug primrose oil) 20.OX Polyethylene glycol 400 monostearate 10.OX Vater to 100.OX * Sucralfate provided by Cuillni Chemie, W. Germany.

The vegetable oil, lanoline and polyethylene glycol 400 monostearate were melted and thoroughly mixed with the warmed water to form an 15 ointment. The sucralfate suspension was incorporated into the ointment.

EXAMPLE 3 A lotion is prepared from the following ingredients (all percentages are by weight): 2:1 suspension of sucralfate'* in polyethylene 15.OX glycol 400 (10X sucralfate in the final product) Glycerol 10.OX Ethanol 50.OX 25 Vater to 100.OX * Sucralfate provided by Cuillni Chemie, W. Germany. 328S19BAJ»l/A2d/OP/MJ/K]»J/t9.M.1990 23 5 6 8 6 is The Ingredients were mixed together In the order stated to obtain a lotion. example 4 A hair tonic is prepared from the following ingredients (all per-5 centages are by weight): Sodium sucrose sulfate 3.OX Ethanol 25.OX Water to 100.OX Sodlua sucrose sulfate was dissolved in wacer and ethanol was then 10 added to the solution.

EXAMPLE 5 A hair tonic is prepared as. described in Example 4 from che following ingredients (all percentages are by weight): Sodium sucrose sulfate 1.5X Ethanol 40.OX Water to 100.OX EXAMPLE 6 A lotion is prepared from the following ingredients (all percentages are by weight): Sodium sucrose sulfate 0.5Z Isciropanol 10.OX Echanol to 100.OX 328t)9BAJ0I/A26/Or/MJ/XPJ/l».04.1990 235 6 86 16 EXAMPLE 7 A cream is prepared from the following ingredients (all percentages are by weight): Sodium sucrose sulfate 2.OX Cetyl alcohol 6.OX Mineral oil 4.OX Paraffin 2.OX Triethanolamine 0,5X Xanthan gun 0.5X 10 Water to 100,OX example 8 A lotion is prepared from the following ingredients (all percentages are by weight): Sucralfate 5.OX Hydroxyethyl cellulose 0.5X Ethanol 25.OX Propylene glycol 40.OX Vater Co 100.OX EXAMPLE 9 A solution is prepared from the following ingredients (all percen tages are by weight): Sodium sucrose sulfate 3.OX Water 50X } Ethanol 25X ) to 100.0% Propylene glycol 25Z ] 3288I9DA.001/A26/OP/MJ/KPJ/19.04.I990 235 686 17 EXAMPLE 10 A shampoo is prepared from the following ingredients (all percentages are by weight): Propylene glycol oleate (PEG-55) 1.5X Sodium lauryl sulfate 28X 23. OX Cocamidopropyl bethaine 9. OX Dimethicone copolyol 1.5X Sodium sucrose octasulfate 3.OX NaCl q.s.

Vater ad 100X Phenopip (preservative) 0.5X EXAMPLE 11 A mousse-conditioner is prepared from the following ingredients (all percentages are by weight): IS Glyceryl stearate (PEG-100 stearate) 1.5X Cetaryl alcohol 1.5X Dimethicone copolyol 0.5X Demineralized water ad 100X Phenopip (preservative) 0.5X Sodium sucrose octasulfate 2.0Z 95X of the above mixture was placed in a spray bottle together with 52 of butane.

EXAMPLE 12 A skin lotion is prepared from the following ingredients (all per-25 centages are by weight): 328819BAJX)1/A26/OP/MJ/KPJ/19JM.1990 18 Clre de lanol "seppic CTO" 4. 21 Macrogoli stearas 400 "Simulsol M 45" 1. 8X Arachidis oleum . 5X Isopropyli myristas 4. 8X Aqua purlficata 68.

OX Sucralfate, micronized (<5 micron) 7.

OX Macrogolum 400 3. 5X Glycerolum 85X 4. 2X Dil. parsoxib^nzcan 10X 1.

OX EXAMPLE li A cream Is prepared from the following ingredients: 235 6 86 Paraffinum solidum Cetanoleum Cetomacrogolum 1000 IS Vaselinum album Acid, ciericum monohydricum Natrll citras Propyleneglycolum Aqua purificaca 20 Sucralfate, micronized (< 5 micron) 19.6 mg 196.0 mg 196.0 mg 588.0 mg 12.3 mg 32.5 mg 56.0 mg 1504.0 mg 196.0 mg EXAMPLE 14 Effect of a treatment with sucralfate on common male-pattern baldness The preparaclon of Example 2 was used in two dally topical applications by a 40-year-old male who had a normal, common male-pattern 25 baldness. After 3-4 weeks, there was appearance of hair in che otherwise bald lateral-frontal areas in the scalp. At the beginning, a "plume" appeared in che area, and after a few days che "plume" began Curning inco real hairs and 8 days after, chere were dozens of real hairs which were indiscinguishable from ocher hairs of che scalp, 30 aparc from being shorter and all coloured in Che original colour of XZSS19HA-00I/AM/OP/MJ/KPJ/19.04.1990 235 686 19 the hair, In contrast to the existing hair, which was partially grayed.

EXAMPLE 15 Clinical evaluation of sodium sucrose sulfate in the treatment of 5 alopecia. A pilot study During a 11 month period (April 1989 to March 1990), a total of 50 patients participated in a pilot study, evaluating the effect of the sodium sale of sucrose sulfate administered topically for the treatment of alopecia.

The test preparation was a 3X solution of Na-sucrose sulfate in a vehicle of 502 water/25X ethanol/25X propylene glycol (w/w), dispensed in 35 ml bottles with a sponge for topical administration on the scalp. The cest preparation was applied morning and evening on the scalp, preferably on areas of partial or total alopecia. In 15 average the patients have been using one bottle per month.

All patients were recruited by one dermatologist, and the effect of treatment was evaluated clinically by the same investigator, at monthly clinic visits.

The clinical effect was classified as vary good in cases of definite 20 cosmetic beneflc, as good in cases of significant regrowth of new hair, but not to an extend chat Is cosmetically apparent at distance, and as no effect in cases of no or only marginal regrowch of hair.

Male androgenic alopecia was the clinical diagnosis in 35 patients, Cheir age ranging from 17 years to 50 years. Period of treatment was 25 about 3 months In 15 case, about 6 months in 10 cases, and about 9 months in 10 cases.

The following results were obtained: 32S8I9BA.001/A26/OF/MJ/KFJ/19.04.1990 235686 Age group (years) Clinical effecc No of Very paCienCs good Cood No effecc Drop-ouCs 17-25 25-40 >40 10 5 0 2 1 4 PaCienCs in the age group 17-25 had previously failed on treatment with minoxidil (Regain®) and showed no effecc on Che tesc preparaclon. 4 PaCienCs in Che age group 25-40 had previously failed on treatment wich minoxidil (Regain®) and all 4 showed a good effecc on che CesC drug.

About 80% of Che male androgenic alopecia suffered from milder to more severe seborrhea in the scalp, and both before and during che CesC drug period Chey received simulcaneous treatment wich Copical sCeroid (becameCasone) and car preparacions. 2 Patients suffered from subtotal or tocal alopecia, wich underlying atopic dermaticis. The first was a 40 year male who had been bald for 20 years, and after CreaCmenC wich Che Cest drug for 10 months, complete regrowch of hair was observed. The second paclent was a 17 year old male wich 2-3 years of alopecia, and CreaCmenC for 8 months wich Che CesC drug, showed only liCCle effecc.

Females aged from 20-49 years wich Ludwig type alopecia were treated with Che CesC drug for 6-8 months. The effect was good in 3 patients, and in 2 pacients Chere was only a marginal effecc, boch showing a better response on previous CreaCmenC wich minoxidil (Regain®). 5 Female paciencs wich alopecia areata aged from 39 years co 46 years were creaced for periods from 2 Co 9 monchs, the CreaCmenC showing a very good effect. 328819BA.001/A26/OP/MJ/KFJ/19.04.1990 t> t> a 6 21 One female 46 years old with very bad hair condition and underlying psoriasis was treated for 9 months and a very good effect on the test drug was obtained. 2 female patients aged 34 and 40 years with underlying discoid LE 5 were successfully treated for 2 and 4 months, respectively, with a very good effect comprising regrowth of hair except for the very center of the lesion.

In conclusion, topical application of a solution comprising the sodium salt of sucrose sulfate (31 solution of sodium sucrose sulfate in 10 a water/ethanol/propylene glycol vehicle), the solution being administered every morning and evening for time periods of 3-11 months showed to be effective in the treatment of male, androgenic alopecia and the Ludwig type alopecia in females. Furthermore, the drug was effective in the treatment of subtotal, total and aerate alopecia in 15 patients with underlying diseases such as atopic dermatitis, psoriasis and discoid LE.

The hair-growing effect in the patients responding to the treatment was typically apparent after 1 month of treatment and in comparison to treatment with topically applied minoxidil (Regain®), the test 20 preparation gave a markedly better effect.

EXAMPLE 16 Clinical evaluation of che prophylactic and t/ierapeueical effect of sodlua sucrose sulfate in che treatment of hair loss secondary co and-cancer treatment. A pilot study The test preparation containing sodium sucrose sulfate from Example 11 was used both prophylactlcally and therapeutically in a few patients undergoing radiation and/or cytostatic antl-cancer treatment. One female started the treatment with the test preparation immediately after the anti-cancer treatment was instituted, applying the test 30 preparation twice daily on eyebrows and eyelashes on the right side, and the left side served as untreated control. The treatment lasted 328819BA.001/A26/OF/MJ/KPJ/19-04.1990 235 6 80 22 four weeks. On che treated side, eyebrows and eyelashes remained but were lost on the untreated side. Another four females - also on anticancer treatment - used the test preparation twice daily, topically on the scalp after the hair was lost. A much faster regrowch of hair 5 than normally was observed in the four females.

In conclusion, topical application of sodium sucrose sulfate as a 3X w/w solution is effective in preventing hair loss secondary to anticancer treatment and, furthermore, sodiuis sucrose sulfate stimulates hair regrowth after anti-cancer treatment. In addition, the results LO show that topically applied sodium sucrose sulfate is of beneficial use for cosmetic purposes such as for the prevention of hair loss.

EXAMPLE 17 Clinical evaluation of sucralfate in the treatment of alopecia. A pilot study A composition containing mlcronized sucralfate (<5 jum, Giulini Chemie, Germany) 5X, ethanol 25X, propylene glycol 25X, and distilled water 45X, was used in two dally applications on the scalp, the composition being dispensed in a bottle with sponge applicator. The average consumption of the test preparation in individual patients 20 was about one bottle per month. One male aged about 55 years, and with male androgenic alopecia Hamilton grade 3, used the test preparation for two months, and there was a dense regrowth of small terminal hairs in the partly balded area of the scalp. The effects were lasting after stopping the treatment for about 4 months. Another 25 six male patients aged from 30 to 48 years, and with male androgenic alopecia Hamilton grade 3 and 4, have used the test preparation for periods of 3 months to 6 months, and in all six cases there have been a definite regrowth of terminal hairs; the effect was typically seen after 6-8 weeks of treatment. In one patient aged 50 and with total 30 alopecia for the last 20 years, a dense regrowth of fine short terminal hairs all over the scalp was observed after application of the test preparation for 6 weeks. 328819BA.001/A26/OP/MJ/KPJ/I9.W.1990

Claims (31)

23 5 6 8 6 23 Ic can be concluded Chat topical application twice dally on the scalp of 5X mlcronized sucralfate suspension In a water/ethanol/propylene glycol vehicle is effective in Che treatment of male androgenic alopecia, also including middle aged patients. 328819BA.001/A26/OP/MJ/KPJ/19.04.1990 235 6 8 6 WHAT//WE CLAIM IS; GSttHS 24

1. A method of treating and/or preventing alopecia, the method comprising administering to a patient in need thereof a therapeutically or prophylactically effective amount of a sulfated mono-, dl- or 5 oligosaccharide or a derivative, salt or complex thereof.

2. A method according co claim 1, wherein the sulfated mono-, di- or oligosaccharide is polysulfated.

3. A method according to claim 1 or 2, wherein the monosaccharide is selected from the group consisting of xylose, fructose and glucose. 10

4. A method according to claim 1 or 2, wherein the disaccharide is selected from the group consisting of sucrose, lactose, maltose and cellobiose.

5. A method according to claim 4, wherein the disaccharide is sucrose, a sucrose derivative, or a complex or salt thereof. 15

6. A method according to claim 5, wherein che disaccharide Is at least tetrasulfeted.

7. A method according to claim 6, wherein che polysulfated disaccharide Is selected from the group consisting of sucrose pentasul-fate, sucrose hexasulface, sucrose heptasulfate and sucrose octasul-

8. A method according to claim 1, wherein the sulfated mono-, di- or oligosaccharide is complexed wich or forms a salt with a metal selected from the group consisting of an alkali or alkaline earth metal, 25 wich an organic base.

9. A method according to claim 5, wherein the polysulfated sucrose is in the form of a potassium or sodium salt. 20 face Al, Zn, Cu, Zr, Ti, Bi, Mn and Oa, or f o • 328819BA.001/A24/OP/MJ/KPJ/19.04.1990 I a SJ <> 25

10. A method according to claim 5, wherein Che complex of che sulfaced sucrose is sucralfaCe. Wv

11. A method according Co any one of the preceding claims, wherein Che sulfaced mono-, dl- or oligosaccharide is combined wich a glucosami-noglycan selected from Che group consisting of hyaluronic acid, dermacan sulfaCe, chondroitin sulfate, keratan sulface, heparan, and heparan sulface. 10 15

12. A method according to any one of the preceding claims, wherein the sulfaced mono-, di- or oligosaccharide is applied topically in a therapeutically and/or prophylactlcally effective dose in a suitable formulation or from a comb or brush, Che sulfaced mono-, di- or oligosaccharide being incorporaced into a vehicle or release system in Che form of a powder or solucion, or in Che form of liposomes or micelles, and in concenCraCions of substantially 0.001-99% w/w. 20

13. A method according to any one of claims 1-11, wherein the sulfated mono-, dl- or oligosaccharide is injected or otherwise introduced parencerally in a suitable formulation or implanc, inco or under che skin or scalp, in a therapeutically and/or prophylactlcally effective dose in 25 concentrations of substantially 0.001-99% w/w.

14. Use of a sulfated mono-, di- or oligosaccharide or a derivative, sale or complex chereof for the preparaclon of a composlcion for 30 CreaCing and/or preventing alopecia.

15. Use according to claim 14, wherein the sulfated mono-, di- or oligosaccharide is polysulfatad. 32S819BA.001/A26/OP/MJ/KPJ/19.04.1990 >" *2 FEB 1993 23 5 6 86 26

16. Use according Co claim 14 or 15, wherein the monosaccharide is selected from che group consisting of xylose, fructose and glucose.

17. Use according to claim 14 or 15, wherein the disaccharide is selected from the group consisting of sucrose, lactose, maltose and 5 cellobiose.

18. Use according to claim 17, wherein the disaccharide is sucrose, a sucrose derivative, or a complex or salt thereof.

19. Use according to claim 18, wherein the disaccharide is at least tetrasulfated. 10

20. Use according to claim 19, wherein the polysulfated disaccharide is selected from the group consisting of sucrose pentasulfate, sucrose hexasulfate, sucrose heptasulfate and sucrose octasulfate.

21. Use according to claim 14, wherein the sulfated mono-, di- or oligosaccharide is complexed with or forms a salt with a metal selec-15 ted from the group consisting of an alkali or alkaline earth metal> with an organic base.

22. Use according to claim 18, wherein the polysulfated sucrose is in the form of a potassium or sodium salt. 20

23. Use according to claim 18, wherein the complex of sulfated sucrose is sucralfate.

24. Use according to any one of claims 14-23, wherein the sulfated mono-, di- or oligosaccharide is combined with a glucosaminoglycan selected from che group consisting of hyaluronic acid, dermatan sulfate, 25 chondroitin sulfate, keratan sulfate, heparan, and heparan sulfate.

25. UBe in the treatment of alopecia of a composition prepared according to any one of claims 14-24, wherein the sulfated mono-, di- or oligosaccharide is applied topically in a therapeutically and/or prophylactlcally effective dose in a suitable formulation or 328819BiV001/A26/OP/MJ/KPJ/19.04.1990 v fc N ; _V» c Al, Zn, Cu, Zr, Ti, Bi, Mn and Os, or \J X. o „ c 23 5 6 8 6 27 from a comb or brush, the sulfated Bono-, dl- or oligosaccharide being incorporated into a vehicle or release system in the form of a powder or solution, or in the form of liposomes or micelles, and in concentrations of substantially 0.001-99% w/w.

26. Use in the treatment of alopecia of a composition prepared according to any one of claims 14-24, wherein the sulfated mono-, di or oligosaccharide is injected or otherwise introduced parenterally in a suitable formulation or implant, into or under the skin or scalp, in a therapeutically and/or prophylactlcally effective dose in concentrations of substantially 0.001-99% w/w.

27. Use of a sulfated mono-, di- or oligosaccharide for combating or preventing hair loss and/or preserving the natural colour of the hair.

28. Use of a sulfated mono-, di- or oligosaccharide for the manufacture of a composition for combating or preventing hair loss and/or preserving the natural colour of the hair.

29. A method as defined in claim 1 of treating and/or preventing alopecia substantially as herein described with reference to any example thereof.

30. Ose as defined in claim 14 or claim 28 of a sulfated mono-, di- or oligosaccharide substantially as herein described with reference to any example thereof.

31. Use as defined in claim 27 of a sulfated mono-, di- or oligosaccharide substantially as herein described with reference to any example thereof. VLH HeTD>TEg- f\/s By the authorised agents \ J PARK & SON n p - •• f