PHARMACEUTICAL AND VETERINARY COMPOSITIONS

Abstract: Pharmaceutic, veterinary and cosmetic compositions used for treating the human or animal body consist essentially of hydroxyalkylphosphine compounds such as tetrakis (hydroxymethyl) phosphonium sulphate and a pharmaceutically, veterinarally or cosmetically acceptable diluent or carrier.

Description: This invention relates to the pharmaceutical and veterinary compositions of hydroxyalkyl phosphines and their derivatives and especially to their use for topical application to humans and animals to cure, prevent or alleviate fungal, bacterial and other microbial infections of the skin, or to alleviate skin odour. In our application E.P. 0139404 A1 we have described the use of certain hydroxyalkyl phosphines for water treatment and in our unpublished application GB 8527793 we have described horticultural and agricultural uses of hydroxyalkyl phosphines to control mosses lichens and plant pathogens. We have now discovered that hydroxyalkyl phosphines are effective against a wide range of fungal, bacterial, protozoal and other microbial infections of humans and animals. Our invention provides a therapeutic or prophylactic composition comprising a hydroxyalkylphosphine compound of the formula (HORPR¹ n ) x X y wherein R is an alkylene group having from 1 to 4 carbon atoms and each R¹ may be the same or different and is an alkyl group having from 1 to 4 carbon atoms or an ROH group, X is an anion such that the compound is at least sparingly soluble in water, n is 2 or 3 and y is 0 or 1 such that (n+y) is 2 or 4; or an at least sparingly water soluble condensate thereof; and a pharmaceutically or veterinarily acceptable carrier or diluent. Preferably the hydroxyalkyl phosphine compound is tris (hydroxymethyl) phosphine or a precursor or most preferably a tetrakis (hydroxymethyl) phosphonium salt. Particularly preferred are tetrakis (hydroxymethyl) phosphonium chloride, sulphate, bromide and phosphate. However X may be any compatible anion such as nitrate, fluoride, a phosphonate such as acetodiphosphonate, aminotris(methylenephosphonate), ethylenediamine tetrakis (methylenephosphonate) or diethylene triamine pentakis (methylene phosphonate), a condensed phosphate such as pyrophosphate, metaphosphate, tripolyphosphate or tetraphosphate, chlorate, chlorite, nitrite, sulphite, phosphite, hypophosphite, iodide, borate, metaborate, pyroborate, fluoborate or carbonate or an organic anion such as formate, acetate, benzoate, citrate, tartrate, lactate, propionate, butyrate, ethylene diamine tetracetate, paratoluene sulphonate, benzene sulphonate or a surfactant anion such as an alkyl benzene sulphonate, alkyl sulphate or alkyl ether sulphate. The hydroxyalkyl phosphine compound may alternatively contain 2 or more phosphorus atoms, and preferably the phosphorus compound is water soluble to a concentration of at least 10 ppm. more preferably at least 20 ppm. most preferably at least 100 ppm e.g. at least 0.5 g/l at 25°C. Such phosphorus compounds contain at least 1 hydroxyalkyl group, usually per phosphorus atom, and preferably at least 2 hydroxyalkyl groups per phosphorus atom. Such hydroxyalkyl groups are preferably of formula ROH, where R is as defined above. The group or groups joining the phosphorus atoms together may be of formula -R-O, -R-O-R or -R-NH-R or -R-R"-R- where R is as defined above and R" is the residue formed by removal of two hydrogen atoms, bonded to nitrogen, from a di or polyamide or di or poly amine, such as urea, dicyandiamide, thiourea or guanidine. Such compounds with 2 or more, e.g. 3, hydroxyalkyl groups per phosphorus atom may be made by self condensation of compounds with 3 or 4 hydroxyalkyl groups attached to one phosphorus atom, e.g. of formula [HOR P R' n O m ] y X x or with a compound of the formula R"H₂ such as urea. Condensation may be performed by heating at 40-120°C. The hydroxyalkyl phosphine compounds are highly effective against fungi such as Trichophyton rubrum, T. mentagrophytes, T. interdigitale and Microsporum canis which causes Athletes Foot, and/or Ringworm infections of humans. The compounds are also very effective against various Streptococci such as S. pyogenes, which is responsible for a variety of human and animal diseases including Impetigo, Erysipalus, Scarlet Fever, Rheumatic Fever and Tonsilitis and other throat infections in humans and Mastitis of cattle, and S. Faecalis which is responsible for urinary infections such as Cystitis. Other bacteria against which the compounds are effective include Proteus species such as P. mirabilis, P. vulgaris which cause urinary infections and pneumonia, Klebsiella pneumonia, which causes pneumonia, and womb infections of mares, salmonella typhimurium and S. Enteritides which cause food poisoning in humans and poultry, Shigella boydii which causes Bacillary Dysentery, Bacillus cereus which causes food poisoning and Legionella pneumophila which causes Legionaires Disease. The compounds are also effective against Candida albicans and Candida tropicalis, which cause Thrush, Sporotrichum schenckii which causes abscessed in humans and horses, Aspergillus fumigatus which causes pulmonary disease and pathogenic protozoa such as Naegleria or Acanthamoeba which cause Amoebic Meningitis. The composition according to the invention is applicable, for example, to the treatment of fungal infections such as Athletes Foot or Tinea (ringworm) and bacterial infections such as Boils, Erysipalus or Impetigo or to reduce foot order, or for the treatment of Mastitis in cattle. The hydroxyalkyl phosphine compounds are especially useful in antiseptic/fungicidal creams, ointments, pastes, collodions, lotions, liniments and powders for treatment of Athletes Foot, Ringworm, Boils and other infections and inflammations of the skin, in foot sprays, and in veterinary dips and udder washes. The compounds may also be used in mouth washes gargles for treating such condition as gum boils, Streptococcal infections and Oral Thrush. They are useful in pessaries and other preparations for treatment of Cystitis, Vaginal Thrush and other urinary and vaginal infections. They may also be useful in topical, parenteral or oral preparations, for treatment, prevention and/or control of a variety of human and animal infections such as Salmonella infections in poultry and humans, Legionaires Disease, various types of Pneumonia, Staphylococcal infections, Rheumatic Fever, Plague, Scarlet Fever, Tuberculosis, Leprosy, Anthrax, protozoal infections, such as Amoebic Meningitis, Amoebic Dysentry, Malaria and Trypanosomiasis, vermal infections such as Hookworm, Filariasis, Toxocariasis, Bilharzia, Ascariasis, Tapeworm and Liver Fluke, and systemic fungal infections such as histoplasmosis, blastomycosis, paracoccidiomycosis, coccidiomycosis, and actinomycosis. The composition may comprise an aqueous or organic solvent such as sterile water, saline solution, ethanol, isopropanol or glycerol, or mixtures thereof or a hydrocarbon, glyceride or other oil, preferably emulsified with water or an aqueous based solvent. Particularly preferred are compositions adapted for topical application containing the hydroxyalkyl phosphine compound carried in a cream or ointment base or absorbed on or absorbed in an inert particulate solid such as talc, kaolinite, bentonite, diatomaceous earth, zeolite or other absorbent aluminosilicate or calcium carbonate. Alternatively the compounds may be encapsulated or micro capsulated in a pharmaceutically acceptable encapsulant such as sugar, fat, gelatin, resin or gum, such as gum acacia, gum arabic, gum tragacanth or a slowly soluble synthetic polymer, such as polyvinylalcohol or absorbed in a slow release of glass such as a phosphate glass. Alternatively the compounds or solutions thereof may respectively be dissolved in or emulsified with liquified gaseous solvents in pressurised containers. The invention also provides medicated shampoos and medicated soap in bar, liquid crystal or liquid form containing the hydroxyalkyl phosphine compounds. The compounds may be tabletted with suitable solids such as chalk, kaolin, sugar or salt or dissolved in syrups, linctus or saline drips. The compositions may additionally contain other fungicides, bactericides or synergists, antiperspirants, lanolin or other skin softening or moisturising preparations, analgesics, antibiotics, emulsifiers, dispersants, carboxymethyl cellulose, soaps, surfactants, polymeric thickening agents, wetting agents, foam controlling agents, perfumes, flavourings, colouring, deodorants, reodorants and antiseptics. A typical ointment, for instance, may contain soft paraffin, petroleum jelly, emulsifying wax, and/or wool fat, optionally in admixture with liquid paraffin and/or hard paraffin, a cream may typically comprise an aqueous based emulsion of emulsifying wax, together optionally with soft paraffin or other hydrocarbons, and/or wool fat. Creams may optionally contain buffers, such as citric acid/sodium phosphate. Ointments or creams may be combined with e.g. zinc oxide or calamine to form pastes. The effective dose depends upon the nature and site of the infection. Typically the hydroxyalkylphosphine compounds exhibit bacterial and fungicidal activity at concentrations within the range 1 to 2000 ppm, especially 5 to 1000, preferably 10 to 500 e.g. 15 to 200 ppm. It is preferred to administer doses adapted to provide such concentrations at the site of infection, subject to toxicological constraints. Compositions according to the invention typically contain from 0.2 to 20% by weight of the hydroxyalkylphosphine compound preferably 0.5 to 10% especially 1 to 3% by weight. Aqueous solutions up to 80% by weight concentration are obtainable, but these normally require some dilution to provide compositions suitable to be administered to humans or animals. Our invention includes concentrates for dilution to provide pharmaceutical, veterinary or cosmetic preparations. THP salts are generally non-carcinogenic and non-mutagenic, but exhibit toxic effects especially on liver cells. Their toxicity in topical applications is generally low, but care must be taken in administering them internally. The invention is of particular value for the treatment of vertebrates including: mammals such as cats, dogs and other carnivores, horses, sheep, pigs, cattle, goats, camels, deer and other ungulates, mice, hamsters, gerbils and other rodents, man and other primates; birds such as chickens, geese, ducks, turkeys and cage birds; reptiles; and fish. The invention will be illustrated by the following examples. Example 1 In the following Tables 2 to 21 the material D denotes a 75% by weight aqueous solution of tetrakis(hydroxymethyl) phosphonium sulphate and Kathon WT® is a registered Trade Mark and represents a widely used proprietory biocide which is an aqueous solution of a mixture of 2-methyl-4-isothiazolin-3-one and 5-chloro-2-methyl-4-isothiazolin-3-one. The results show that material D has a high degree of lethal activity to bacteria within 2 to 6 hours of exposure at concentrations of at least 20 ppm and a high degree of lethal activity to the fungi at 100-150 ppm after 24 hours. TABLE 1 (a) Bacteria The following bacterial strains were used in the tests: Streptococcus pyogenes NCTC 8198 Streptococcus faecalis ATCC 19433 Sarcina lutea ATCC 9341 Proteus mirabilis NCTC 8559 Proteus vulgaris NCTB 4175 Klebsiella pneumoniae NCTC 10341 Salmonella typhimurium NCTB 10258 Salmonella enteritidis NCTC 6676 Shigella boydii NCTC 9328 Bacillus subtilus ATCC 6633 Bacillus cereus ATCC 11778 (b) Yeasts and fungi The following yeast and fungal strains were used in the tests: Candida albicans ATCC 10231 Candida tropicalis NCPF 3111 Absidia corymbifera NCPF 2001 Sporotrichum schenkii NCPF 3182 Trichophyton rubrum NCPF 197 Trichophyton mentagrophytes NCPF 351 Microsporum Canis NCPF 351 Penicillium funicolosum CMI 114933 Aureobasidium pullulans CMI 45533 Inocula for the bacterial strains were prepared from cultures on Tryptone soy agar (TSA, Oxoid CM 131) incubated at 37°C for 24 hours. Surface growth of the organisms was suspended in sterile physiological saline and diluted with this medium to give 50% transmission at 520 nm on an SP 600 spectrophotometer, an approximate yield of 10⁸ colony forming units (cfu) per ml. For use as inocula these suspensions were further diluted in sterile physiological saline to give an expected fnal concentration in the test solutions of 10⁴ to 10⁵ of cfu/ml. Inocula for the two yeast (Candida) strains wre prepared from cultures on Sabouraud dextrose agar (SDA Oxide CM 141) incubated at 37°C for 48 hours. Surface growth of the organisms was suspended in sterile physiological saline and standardised by plate counts to contain approximatedly 10⁶ cfu/ml. For use as inocula these suspensions were diluted to give an expected final concentration in the test solutions of 10⁴ to 10⁵ cfu/ml. Inocula for the fungal strains were prepared by sub-culturing the organisms from SDA into Sabouraud liquid medium (SLM, Oxoid 147) and incubating at 30°C for 5 to 10 days. Mycelial growth was then broken down by shaking the cultures with glass beads and the resulting suspensions were standardised by plated counts to contain approximately 10⁶ cfu/ml. All tests were carried out in phosphate buffered saline (PBS) prepared from Oxoid tablets (Dulbecco A, Oxoid BR14a) and having the formula: Sodium chloride 8 g/litre Potassium chloride 0.2 g/litre Disodium hydrogen phosphate 115 g/litre Potassium dihydrogen phosphate 0.2 g/litre pH 7.3 g/litre Samples from the inoculated test and control preparations were diluted 1 ml to 9 ml in 0.1% peptons water containing 1% v/v Tween® 80 as an inactivator, and serially diluted in further ten-fold steps in the same dilutent. The media used for preparing plate counts from the diluted samples were: (a) for bacterial strains - Tryptone soy agar containing 0.2% sodium thioglycollate (b) for yeast and fungal strains - Sabouraud dextrose agar containing 0.2% sodium thioglycollate. For preparation of the test series each was diluted in PBS 1 part in 1000 parts to give a stock solution of 1000 ppm. This was freshly prepared on each test day. The stock dilution was then further diluted to give the test series of: (b) 500, 200, 100, 50 and 20 ppm for the organisms tested Strep. pyogenes, Strep. faecalis, Shigella boydii, Salmonella enteritidis and Klebsiella pneumoniae) and: (c) 500, 100, 20, 5 and 1 ppm for the remainder of the test organisms. The test solutions as described in (5) above were prepared in 50 ml volumes and the test solutions plus a 100 ml volume of PBS (as control) were inoculated with appropriate volumes (0.5 to 1 ml) of the inocula prepared as described in (2) above, shaken to evenly distribute the inoculum and held at room temperature. Immediatedly after inoculation (time 0) and then at pre-determined times 0.1 ml aliquots were removed from the test and control solutions for counting. The 0.1 ml samples were serially diluted in hundred-fold steps in the peptons water/Tween® 80 diluent and 1 ml volumes of the dilutions were used to prepare duplicate pour-plates in TSA/sodium thioglycollate medium for the bacteria and Sabouraud/sodium thioglycollate medium for the yeasts and fungi. The bacterial count plates were incubated at 37°C for 24 hours, the yeast (Candida) count plates at 37°C for 48 hours and the fungal count plates at 30°C for 5 to 10 days. Developing colonies were then counted and by reference to the appropriate dilution ratios the numbers of surviving organisms at each compound concentration/time point were calculated. Example 2 The activity of 50% aqueous bis THP phosphite, 50% aqueous mono THP phosphite and 80% aqueous THP sulphate against Trichophyton rubrum were compared in a 7 day test in phosphate buffered saline solution to determine ED₅₀ and ED 90 values. In contrast under the same conditions phosphorous acid, which is widely used as a fungicide, requires concentrations of the order of 0.1 to 0.2% by weight to produce a useful reduction in the growth of T. Rubrum. Example 3 THPS was found to give a complete in vitro kill of Naegleria (the protozoon responsible for Amoebic Meningitis) at concentrations of 15 ppm. Example 4 One of ten Koi Carp, in a fish pond, exhibited symptoms of protozoal infection. 100 ppm of 75% aqueous THP sulphate was dosed to the pond via the oxygenating venturi to provide a concentration of 27 ppm in the pond water. The infected fish recovered fully and none of the other fish became infected. The fish fed normally throughout the treatment and exhibited no signs of distress. Example 5 A cream for treatment of fungal infections of human skin by topical application comprises: 9% emulsifying wax 15% soft white parafin 6% liquid parafin 0.5% citric acid monohydrate 2.5% sodium phosphate 2% THP phosphate balance sterile water When the cream is applied twice daily to areas of ringworm infection full remission is normally observed within 1 to 2 weeks. Example 6 A mouthwash for dilution with an equal volume of warm water prior to use comprises: 1.5gm sodium chloride 1.0gm sodium bicarbonate 2.5ml peppermint emulsion 1ml 75% THP chloride solution 50ml chloroform water balance to 100ml sterile water The composition produces rapid alleviation of oral thrush. Example 7 Nasal drops comprise: 500 mg Ephedrine hydrochloride 500 mg Chlorobutanol 500 mg Sodium Chloride 1ml 75% aqueous THP chloride balance to 100ml water Example 8 Foot powder comprises: 25% zinc oxide 25% starch 50% talc The starch is first mixted with 12%, by weight thereof of 75% aqueous THP sulphate. The powder produces rapid alleviation of Athletes Foot. Example 9 A foot spray comprises the starch preparation of example 8 dispersed in a pressure liquified propellent gas, together with a perfume and an antiperspirant. The composition provides reduction in foot odours and is useful in preventing or alleviating Athletes Foot

Claims: A therapeutic or prophylactic composition comprising a hydroxyalkylphosphine compound of the formula (HORPR¹ n ) x X y wherein R is an alkylene group having from 1 to 4 carbon atoms and each R¹ may be the same or different and is an alkyl group having from 1 to 4 carbon atoms or an ROH group, X is an anion such that the compound is at least sparingly soluble in water, n is 2 or 3 and y is 0 or 1 such that (n+y) is 2 or 4; or an at least sparingly water soluble condensate thereof; and a pharmaceutically or veterinarily acceptable carrier or diluent. Composition according to claim 1 wherein R has 1 carbon atom. Composition according to either of claims 1 and 2 wherein each R¹ has 1 carbon atom. Composition according to any foregoing claim wherein R¹ is R(OH). Composition according to claim 1 wherein the hydroxyalkylphosphine compound is a water soluble tetrakis (hydroxymethyl) phosphonium salt. Composition according to any of claims 1 to 5 which is antiseptic and/or antifungal cream, lotion, paste, collodion or ointment for topical application to the human or animal body. An ointment according to claim 6 comprising liquid paraffin and/or soft paraffin and/or wool fat and/or wool fat derivative. A cream according to claim 6 comprising the composition according to claim 7 dispersed or emulsified in water. Composition according to any of claims 1 to 5 which is a powder for application to the human or animal body. A composition according to claim 9 comprising talc, zinc oxide, and/or starch or a derivative thereof. Composition according to any of claims 1 to 5 which is a pessary. Composition according to any of claims 1 to 5 which is an aerosol spray composition. Composition according to any of claims 1 to 5 which is a soluble capsule. Composition according to any one of claims 1 to 5 which is a tablet. Composition according to any one of claims 1 to 5 which is an aqueous composition for oral or parenteral administration. A mouthwash according to claim 15. Nasal drops according to claim 15. Composition according to any one of claims 1 to 5 which is a soap bar containing at least 0.2% by weight of said hydroxyalkylphosphine compound. Composition according to any one of claims 1 to 5 which is a veterinary dip containing at least 0.2% by weight of said hydroxyalkylphosphine compound. A composition according to any of claims 1 to 19 containing from 0.2% to 20% by weight of said hydroxyalkylphosphine compound. A composition according to claim 20 containing from 1 to 10% by weight of said hydroxyalkyl phosphine compound. A composition according to any of claims 1 to 21 additionally comprising at least one other active ingredient selected from fungicides, bactericides, synergists, antiperspirants, styptics, carboxymethyl cellulose, soaps, surfactants, polymeric thickening agents, wetting agents, foam controlling agents, perfumes, flavouring, colouring, deodorants, reodorants and antiseptics. A concentrate adapted on dilution with water to provide a composition according to any of claims 1 to 22.

Inventors: DAVIS, KEITH PHILIP, SMITH, BRIAN, TALBOT, ROBERT ERIC
Applicants: ALBRIGHT & WILSON LIMITED
Application year: 1988
Main classification: A61K31
IPC codes: A61Q3, A01N57, A61Q11, A61K8, A61K31, C02F1