GB2078109A - Antimicrobial composition - Google Patents

Abstract

Antimicrobial compositions comprise a mineral acid salt of a benzophenanthridine alkaloid and a non-toxic metal salt of an acid in a suitable solvent vehicle. The composition is suitable for the prevention and treatment of infections and diseases in mammals and is particularly in the form of a dentifrice. The alkaloid may be chelerythrine, sanguinarine, protopine and/or homochelidonene and the salt is preferably stannous fluoride, zinc chloride and/or sodium fluoride. <IMAGE>

Description

SPECIFICATION Antimicrobial composition This invention relates to the composition and preparation of an antimicrobial agent which may be used in dental preparations, surgical and other soaps, various other topical preparations, surgical injectable medicines, and other drug applications. In particular, the invention relates to compositions containing mineral acid salts of benzophenanthridine alkaloids mixed with a metal salt, preferably a metal salt of a mineral acid, although salts of mono- or dicarboxylic acids, among others, may also be used.

One of the important sources of sanguinarine is a perennial herb native to North America called Sanguinaria canadensis Li n ne (Family: Papaveraceae) commonly known as blood root, red root, puccoon, etc. The plant contains benzophenanthridine alkaloids including sanguinarine, chelerythrine, and several others. The major alkaloids present are sanguinarine and chelerythrine. The eighth edition of the Merck Index lists the alkaloids as sanguinarine, chelerythrine, protopine and homochlelidonine. The pure chemicals sanguinarine, chelerythrine, and other benzophenanthridine alkaloids can be isolated from other plants besides Sanguinaria. Also, they are available, although very rarely, from some chemical supply houses.Semi-purified forms of the alkaloids are commercially available, and these are generally referred to as sanguinarine nitrate and sanguinarine sulphate. These "salts" are the salts of the mixed alkaloids of the plant Sanguinaria: mainly sanguinarine, chelerythrine, and protopine. While few references can be found in the literature regarding the usage of any of the pure benzophenanthridine alkaloids, plants containing such compounds have been used for medical purposes for quite some time for a wide variety of aliments.

The principal use of sanguinarine until recently was as a stimulant expectorant in cough syrups containing "sanguinarine nitrate".

The use of sanguinarine with thiophosphoric acid in various animal and human neoplasms is shown in French Patents Nos. 70-22029 and 2,152,972.

The alkaloid sanguinarine in solution has been shown to have some antifungal and antiprotozoan properties. The sanguinarine is applied topically as an emulsion to fungal infections. The antibacterial activity of sanguinarine has been found to vary with the attached radicals, and various salts of sanguinarine have been shown to have some activity.

The hydrochloride and the sulphate salts have been found to have some activity against certain bacteria at various concentrations. Sanguinarine nitrate is reported to have some weak bacteriostatic action on various types of bacteria.

The present invention relates to the preparation and the use of antimicrobial agents, formed particularly from mineral acid salts of benzophenanthridine alkaloids and a metal salt and useful in dental preparations, mouthwashes, rinses, surgical soaps, shampoos, creams, lotions, powders, injectables, etc., and other forms of drug preparation and disinfectants. The mineral acid salts of the benzophenanthridine alkaloids may be used in various concentrations with a metal salt as an antimicorbial agent for use in treating both human and animal infections and diseases.

The invention thus provides an antimicrobial composition comprising a mineral acid salt of a benzophenanthridine alkaloid and a non-toxic metal salt of an acid in a suitable solvent vehicle.

The invention also provides a method for preparing an antimicrobial agent comprising: (a) dissolving a benzophenanthridine alkaloid in a mixture of chloroform and methanol; (b) acidifying the solution with a mineral acid to convert the alkaloid to an alkaloid salt; (c) evaporating the acidic solution to dryness; (d) recrystallizing the residue from a mixture of 50% ethanol and 50% chloroform; (e) dissolving the crystals in a solvent to make a solution of at least 0.3% by weight of crystals; and (f) mixing the solution with at least 35% by weight of a metal salt of an acid.

Metal salts particularly useful in the formulations of the present invention include metal salts of halogen acids. Among these salts are zinc chloride, stannous fluoride, and sodium fluoride, although any metal can be used. This includes alkali, alkaline earth, and heavy metal fluorides, chlorides, bromides, and iodides.

Although non-toxic metal salts of halogen acids are preferred for use in the formulations according to the present invention, it has been found that nontoxic salts of other acids, such as mineral acids and mono- and dicarboxylic acids, are also effective. For example, the non-toxic metal salts of sulphuric acid, mitric acid, and acetic acid can be used.

Glycerol is the preferred vehicle of the formulations according to the present invention, although other vehicles that can be used include organic solvents such as propylene glycol, dimethyl sulphoxide (DMSO), lower alcohols, and the like.

The antimicorbial agent of mineral acid salts of a benzophenanthridine alkaloid and a metal salt, according to the invention is useful for topical administration, injectables and other forms of drug preparations. The benzophenanthridine alkaloid salt-metal salt preparation is suitable for treatment of periodontal diseases, prevention of dental caries and similar ortal cavity impairments. The drug preparation of a benzophenanthridine alkaloid and a metal salt is useful for treatment of ringworm infections, acne, cold sores and various parasitic infections. It is also useful for treatment of scours in ani mals.

The drug preparation of the present invention can be incorporated in dental preparations, toothpastes mouth rinses, surgical and other soaps, vehicles for topical applications, vehicles for parenteral or intramuscular injection, and the like.

The antimicrobial agent according to the invention may be prepared as follows. The pure chemical, either sanguinarine, chelerythrine, or other benzophenanthridine alkaloids, is dissolved in a chloroform/methanol mixture and acidified with a mineral acid, such as hydrochloric acid. The acidic mixture is evaporated to dryness and the residue is recrystallized from ethyl alcohol/chloroform, 50/50.

For use, the mineral acid salt of the benzophenanthridine alkaloid is dissolved in either deionized water or C,-Ç6 alcohols, glycerine, propylene glycol, petrolatum, or other organic solvents at 70"C, and a metal salt ora salt-forming acid is added to the above solutions. The preparations generally contain 0.1% byweightand upto 20% by weight of the benzophenanthridine alkaloid salt, and at least 1% and up to 60% by weight of a metal salt, with the remainder being the solvent. The material can be diluted to the desired concentration, depending on the type of use, with the solvents listed above.

The benzophenanthridine alkaloid salt is used in preparations containing 0.01%-10% benzophenanthridine by weight. The metal salt is present in amounts ranging from about 2% to about 60%. The lower concentrations are generally effective in the treatment of most diseases as explained below.

An example of a basic preparation: Sanguinarine chloride 0.3% glycerine U.S.P. 64.7% zinc chloride AR 35.0% The basic preparation can be varied by using, in place of sanguinarine chloride, 0.3% of another mineral acid salt of a benzophenanthridine alkaloid, such as chelerythrine chloride.

A second example of a preparation is: sanguinarine chloride glycerine U.S.P. 96.0% zinc chloride AR 3.0% Athird example of such a preparation is: sanguinarine chloride 1.0% glycerine U.S.P. 64.0% zinc chloride AR 35.0% A fourth example of such a preparation for dental use is: sanguinarine chloride 1.0% glycerine U.S.P. 95.6% zinc chloride AR 3.0% stannous fluoride 0.4% Additional formulations for a basic preparation are as follows: sanguinarine chloride 1.0% stannous fluoride 0.4% and Sanguinarine chloride 1.0% sodium fluoride 3.0% glycerine U.S.P. 96.0% Numerous types of diseases were treated in humans and in animals with the composition of the present invention as follows:: Example 7 1- Canine Ringworm A 5-10% solution of the first basic preparation was used, applied directly to the infected area, one to three applications as indicated, 48 hours apart.

Example 2 -- Feline Ringworm A 48% solution of the first basic preparation was applied directly to the infected area, 48 hours apart, up to three applications.

Example 3-Bovine Ringworm The etiologicai agent of this involvement is usually the mould known as Trychophyton album. The duration of the disease is 4-12 months A 30% dilution of the first basic preparation is used, applied directly to the involved areas, 48 hours apart. Three applications proved to be adequate in treating the condition successfully.

Example 4 - Bovine Neo-natal Diarrhoea Twelve animals having diagnosed and confirmed neo-natal diarrheoa were treated with 0.75 grams of the first basic preparation orally, once per animal, and showed clinical cure with one exception.

This constitutes an excellent result, considering that conventional antibiotic therapy currently in use has a much lower percentage of success.

Sanguinarine chloride and chelerythrine chloride have strong antimicrobial properties. Zinc chloride has antimicrobial properties only in high concentrations. It can be seen from Table I that sanguinarine chloride mixed with zinc chloride in a 1:1 ratio, as a rule, did not show a synergistic effect or even an additive action against most microorganisms tested in vitro. Further, it was found that in most cases the antimicrobial action of the sanguinarine chloride and zinc chloride mixture depended mostly on the amount of sanguinarine chloride present in the mixture, and was relatively independent of the amount of zinc chloride.

TABLEI Mean inactivating dose in micrograms per mil/ilitre {ugiml) of media Microorganism ZnC12 Sanguinarine Zinc12 & Sanguinarine Chloride Chloride (1:1) Bacillus subtilis 25,000 22 1,000 Escherichia coli 6,250 270 500 Klebsiella pneumoniae 3,125 540 1,000 Proteus vulgaris 12,500 590 1,000 Staphylococcus aureus 6,250 70 500 Streptococcus faecalis 25,000 393 500 Streptococcus mutans 1,563 161 63 Candida albicans 150 Saccharomyces cerevisiae 6,250 20 63 Pseudomonas aeruginosa 3,500 7,000 400 A separate test was conducted to determine the inhibitory concentration of sanguinarine chloride alone.These concentrations, for microorganisms in vitro, are as follows: 100 micrograms per millilitre forEscherichia coli 100 micrograms per millilitre for Candida albicans 50 micrograms per millilitre for Streptococcus mutans 10 micrograms per millilitre for Staphylococcus aureus It was further found that a concentration of sanguinarine chloride of 25 micrograms per millilitre caused a 100% reduction of dental plaque by inactivating plaque-forming microorganisms freshly collected from human dental plaque. Sanguinarine chloride compared favourably in vitro to chiorhex- idine (Hibitane (Trade Mark)), a material used as a standard in evaluating inhibition of human dental plaque-forming microorganisms.

However, underin vivo test conditions, sanguinarine chloride was found to be ineffective against plaque-forming microorganisms.

When sanguinarine chloride was applied to the affected area of both dogs and humans, repeated treatment with sanguinarine chloride alone did not reduce dental plaque or alleviate the symptoms of gingivitis or periodontal disease. Continuous treatment with sanguinarine chloride did not prevent the accumulation of dental plaque on teeth or prevent the occurrence of periodontal disease. However, it has been found that a combination of sanguinarine chloride and zinc chloride in glycerine is effective in vivo in reducing dental plaque and the incidence of periodontal disease, and has shown definite promise in the treatment and prevention of human periodontal disease.

Results of tests on guinea pigs with induced ringworm infection, dogs with periodontal disease, and humans with periodontal disease have shown that glycerine preparations of sanguinarine chloride plus zinc chloride are far superior for the management of infections in vivo than either zinc chloride or sanguinarine chloride alone.

When zinc chloride was replaced with a fluoride salt in the present preparations, the activity ofthe preparation against microorganisms believed to be associated with the causation of dental caries and periodontal disease (Streptococcus mutans), as well as other microorganisms, remained identical. Data supporting this phenomenon is presented in the following Tables:: TABLE II Minimum Inhibitory Concentrations (MIC) in ugiml 1. 0% sanguinarine 1.0% sanguinarine AEROBIC chloride chloride MICROORGANISM 3.0% zinc chloride 0.4%stannous TESTED in glycerol fluoride in glycerol Escherichia coli 158 160 Klebsiella pneumoniae 158 160 Proteus vulgaris Streptococcus mutans 79 80 Streptococcus faecalis < 20 < 20 Staphylococcus aureus < 20 < 20 Pseudomonas aeruginosa 630 1,280 Saccharomyces cerevisiae < 20 < 20 Candida albicans 79 < 20 TABLE III Minimum Inhibitory Concentrations (MlC) in ugiml 1. 0% sanguinarine 1. 0% sanguinarine ANAEROBIC chloride chloride MICROORGANISMS 3.0% zinc chloride 0.4% stannous fluoride TESTED in glycerol in glycerol Bacteroides melaninogenicus 8 8 Eikenellacorrodens 32 32 Actinomyces viscous 8 8 Actinobaclllus actinomycetemcomitans 16 16 Capnocytophaga gingivalls 8 8 Capnocytophaga sputigena 8 8 TABLEIV Minimum Bactericidal Concentrations (MBC) in uglml 1.0% sanguinarine 1. 0 /0 sanguinarine chloride chloride MICROORGANISMS 3.0% zinc chloride 0.4%stannous TESTED in glycerol fluoride in glycerol Escherichia coli 158 160 Klebsiella pneumoniae 1,261 640 Proteus vulgaris 2,522 640 Streptococcus mutans 158 160 Streptococcus faecalis 315 160 Staphylococcus aureus 158 80 Pseudomonas aeruginosa 2,522 2,560 Saccharamyces cervisiae 20 20 Candida albicans 79 160 The above phenomenon is not what would be expected in the light of the in vitro test results. In most cases sanguinarine chloride in vivo was only slightly effective or not at all. This was quite unexpected considering that in vitro all the antimicrobial activity of a sanguinarine chloride-metal salt mixture could be explained by the amount of sanguinarine present in the mixture.

Controlled clinical tests on 24 male beagle dogs showed that after four weeks of treatment the dogs treated with the zinc chloride-sanguinarine chloride-glycerine mixture had the lowest plaque and gingivitis scores, while the dogs treated with sanguinarine chloride alone had the highest. The dogs were treated topically once daily with the respective test formulation.

The results presented in Table V clearly indicate that the group of dogs treated with sanguinarine chloride-zinc chloride in glycerine had lower gingivitis scores after four weeks of treatment than did the other groups. Zinc chloride alone was slightly active, but sanguinarine chloride alone showed no activity in vivo at all. This result is unexpected, considering that, in vitro, sanguinarine chloride is quite effective against microorganisms.

TABLE V Oral Clinical Studies -- Beagles (dogs) After Treatment 0(start) 4weeks Mean Gingivitis Scores None 0.475 0.91 0.1% Sanguinarine chloride 0.495 0.942 2.7% Zinc chloride 0.44 0.75 0.1% Sanguinarine chloride with 2.7% zinc chloride in glycerine 0.418 0.548 Mean Plaque Scores None 7.478 12.408 0.1% Sanguinarine chloride 8.885 12.89 2.7% Zinc chloride 8.76 10.15 0.1% sanguinarine chloride with 2.7% zinc chloride in glycerine 8.578 7.407 Mean Pocket Depths None 1.459 1.458 0.1% Sanguinarine chloride 1.415 1.48 2.7% Zinc chloride 1.46 1.37 0.1% Sanguinarine chloride with 2.7% zinc chloride in glycerine 1.445 1.44 Similar results occurred when the dogs were evaluated for the return of dental plaques (See Table Vforplaque scores). Sanguinarine chloride alone was not active in vivo.Zinc chloride was slightly preventive, but the preparation of sanguinarine chloride-zinc chloride in glycerine not only prevented the return and further proliferation of dental plaque but significantly reduced it further during the four weeks of treatment.

Data also indicate that even the pocket depths were reduced somewhat by the treatment materials containing zinc chloride in glycerine, or zinc chloride-sanguinarine chloride in glycerine.

In a clinical test involving twenty volunteer patients with periodontal disease, it was observed that there was rapid improvement after treatment with benzophenanthridine alkaloid chloride-zinc chloride in glycerine. Inflammation, infection and pockets were eliminated, abcesses ceased, gingival tone greatly improved, tissues healed, and in some cases normal tissue was restored and tooth mobility was reduced.

The clinical studies above showed that sanguinarine chloride in vivo was only slightly antimicrobial, very slow acting, or had no effect on the course of the infection at all.

Zinc chloride in the concentrations required in vivo to have antimicrobial action caused blanching oftissue and, in some cases, chemical burns and other tissue damage. Furthermore, zinc chloride was found to be slow acting in vivo as an antimicrobial agent.

Glycerine preparations containing sanguinarine chloride or other benzophenanthridine alkaloids and zinc chloride or stannous fluoride or sodium fluoride were fast acting, requiring only one to three applications to clear out infections rapidly. Furthermore, these preperations did not appear two have the undesirable side effects of the zinc chloride in the concentration necessary to achieve antimicrobial effectiveness.

Example human Periodontal Disease It has been reported that periodontal (gum) disease affects 2 out of 3 middle-aged Americans. Destruction of the tissue and structures that hold teeth fast in their sockets accounts for 75% of tooth loss after the age of 40. Most cases of periodontal disease are the result of neglect and can largely be prevented by a regular programme of thorough hygiene. In the most common type of periodontal disease, the three chief culprits are bacteria, calculus (tartar), and food debris.

The invention has been used by dentists in clinical management of over forty cases of various types of human periodontal disease.

In some cases, even a single treatment brought major improvements in the conditions of the diseased gums. Clinical cure resulting from the treatment was quite apparent and included: elimination of inflammation, normal tissue tone restored, pockets were eliminated, infections were cleared up, mobility was reduced, and gingival tone was greatly improved.

The materials and methods used were the follow ing: 1. Undiluted Paste for Packing: in cases ofwidespreadtissue involvement, undiluted preparation was used in a quantity sufficient (q.s.) to "cover" or"pack" the infected or inflammed areas of gingival tissues.

The clinical procedure consisted of two treatments approximately two weeks apart, with the application of not less than 1.0 mm thickness of the basic preparation to the diseased periodontium. In cases where undiluted material was used, it was either applied topically with a spatula or 0.25 ml was pressed through a 22 gauge needle attached to a 1.0 ml pressure syringe. The material was introduced into the gingivae to the attachment (q.s.) to fill the pocket and the madication was left in place for 10-15 minutes.

2. String Technique: Cotton String Saturated with the Drug Preparation.

In cases where individual teeth were to be treated, an ordinary soft cotton string, sterilized before use, or "gingipak" was used "gingipak contains racemic epinephrine hydrochloride 8-100 solution and 1% benzyl alcohol as preservative).

Cotton strings 1-1.5 cm in length were saturated with undiluted material by using a spatula and precut pieces of string which were then impregnated on a dentist's mixing pad. These strings, when properly impregnated with the preparation, weigh approximately 35 mg/cm.

One to three strings were used per tooth depending upon the circumference of the tooth. Up to three strings were sometimes used for a total of approximately 10.5 mg of the preparation. Impregnated strings were left in place 10-15 minutes.

Dental floss or similar substrate, such as synthetic hollow fibre string, can be impregnated with the basic preparation for use in treating the teeth and gums.

3. Dilution of Paste for Irrigation In addition to "packing" ofthe periodontium or employing the "string technique" on individual teeth, irrigation was often used concurrently as part of the regimen. Generally, the diseased periodontium was first packed with undiluted preparation or the teeth were individually treated by the string technique described above. These methods of treatment were followed by irrigation with a suspension of the preparation in either water or glycerine.

Suspensions were prepared to contain 1 part of full strength material to 1 part of glycerine, or 1 part of water depending on whether a glycerine or water suspension was desired. The final suspension was a V/V mixture of 1 part of material to 1 part of diluent.

Irrigation was accomplished by filling a 7.0 ml syringe to contain 420 mg of the suspension. For treatment of individual teeth, a total of 1.0 ml of material in suspension was used to irrigate the buccal, lingual, and interproximal areas about the teeth.

Where indicated, all teeth were irrigated with 840 mg of the suspension contained in two syringes.

Example 6-Dental Caries Preparations containing about 0.3% sanguinarine chloride and about 35% zinc chloride were used on seven patients with dental caries. Decay was removed from the teeth with a spoon excavator leav ing a layer of carious tissue about 1 mm to 12 mm in depth. The antimicrobial preparation was placed over the remaining decay, about 56.1 mg of preparation, with a Hollenbeck carver and uniformly applied over the decayed area with a piece of cotton held in cotton tweezers. Intermediate restorative material (IRM) was used as a temporary restoration to seal the material in the cavity preparation.

After several weeks (6 weeks) specimens for bacteriological and histological studies, including electron microscopy, were taken.

The conclusions of the investigators were that the preparation could be considered a cariostatic agent.

Furthermore, the material may enhance sclerotic dentine formation, thus forming a hard protective floor between the carious lesion and the pulp.

The benzophenanthridine chloride-zinc chlorideglycerine composition can be used in conjunction with a fluoride-providing compound. These compounds are characterized by the ability to release fluoride ions in water and by substantiai freedom from reaction with other compounds present in the oral preparation. Among these materials are inorganic fluoride salts such as suitable alkali metal, alkaline earth metal, and heavy metal salts. Alkali metal and tin fluorides, such as sodium and stannous fluorides, and mixtures thereof, are preferred.

The amount of the fluoride-providing compound is dependent to some extent upon the type of compound, its solubility, and the type of oral preparation, but it must be a non-toxic amount. Any suitable minimum amount of such a compound may be used, but it is preferably to employ sufficient compound to release from 0.005% to 1%, most preferably about 0.1%, by weight of fluoride ion. Typically, in the cases of alkali metal fluoride and stannous fluoride, this component is present in an amount up to 3% by weight, based on the weight of the preparation, and preferably in the range of from 0.05% to 1%.

Example 7- Treatment of animal Skin Tumours Equine sarcoid (a locally malignant connective tissue tumour found on the skin of horses, similar to fibrosarcomas which occur in other animals and man) is probably of viral origin and transmissible between horses. These tumours are invasive and commonly result in sufficient debilitation of the horse that euthanasia is performed.

Surgical removal is the accepted means of therapy. Recurrence ofthetumour is the rule rather than the exception.

Sixty tumours were treated with the preparation employing the following treatment method: The first basic preparation is either spread on the tumour with a wooden applicator, bandaged and observed in 24 hour intervals, or about 3 mm thickness of the preparation is placed on a Telfa (Trade Mark) pad covering an area slightly larger than the base area of the tumour, and the Telfa pad is secured on the tumour with a bandage. Observation is made in 24 hour intervals. The application is repated if needed after 48 hours.

Three to four or more applications may be necessary in cases of long duration and if the tumour is deep. Recurrence of the tumour occurred in only 25% of the cases.

In all cases, the tumours were removed by the excising action of the preparation and in no case thus treated did a post treatment infection (secondary infection) develop. This was true despite the fact that no covering was maintained on the lesions after the tumour came out, nor was any other form of antibacterial medication applied.

After the second orthird application, the tumour sloughed out, a very firm scab formed on the tumour base, and healing proceeded under the scab. The healing of the wound was slow, but uneventful.

Scars remaining were quite small.

Example 8-Equine Dermal Fibrosa rcomas Seventy-five cases of equine dermal fibrosarcomas were treated topically as described in Exam ple 7. Only 20% of the tu mours recurred within the one year observation period While the cure rate is somewhat less than 80%, this compares favourably with the approximate 55 h cure rate of this tumour following surgical removal and radiotherapy.

Example 9 - Bovine Squamous Cell Carcinoma Seven Hereford cattle diagnosed with the disease were treated with about 15 grams of the first basic preparation pasted right over the lesions. Improve mentofclinical condition followed in all cases.

Example 10- TreatmentofSquamous Cell and Basal Cell Carcinoma in Animals of Non-Bovine Species Canine - Histologically confirmed squamous cell carcinoma of both eyes; five applications with the first basic preparation in five days; in two weeks time, tumours reduced in size and were sloughing.

Equine - Histologically confirmed basal cell carcinoma was treated by six direct topical applications of the preparation locally, once daily. In three weeks, most visible tumours were gone.

Equine - Histologically confirmed large hair follicle carcinoma on chest; tumour removed by surgery; ten topical applications of the preparations into the open wound. In three months lesion heaied and no recurrence of the tumour was noted.

Example 11 - Treatment of Human Skin Tumours Basal Cell Epithelioma: Aterm of qualified dermatologists and surgeons treated sixty patients with diagnosed histologically confirmed basal cell epithelioma (basal cell carcinoma).

The first basic preparation was directly applied topically to the involved area and held in place with a bandage. The lesions were observed in 48 hour intervals and the preparation was required if it was indicated. In most cases, 2-3 applications were adequate. Clinical cure followed the treatment in all cases and no reappearance of the tumours was noted over a four year period.

Example 12-Squamous Cell Carcinoma Sixteen cases of histologically confirmed squanmous cell carcinoma were treated by the same method used in Example 11. In all treated cases, clinical cure followed treatment and only in one case was tumour recurrence noted within four years.

Example 13 - AntimicrobialActivity In order to illustrate the antimicrobial effective ness of the composition according to the present invention, a series of in vitro and in vivo tests were performed. These tests were designed to demonstrate the lowest concentration of sanguinarine chloride that will inactivate microorganisms.The findings were as follows: On the average a 22 ug/ml sanguinarine chloride solution will inactivate cultures of the bacteria Bacillus subtilis, a 270 ug/ml solution cultures of Escherichia coli, a 540 ug/ml solution cultures of Klebsiellapneumoniae, a 590 ug/ml solution cultures ofProteus vulgaris, a 70 ug/ml solution cultures of Staphylococcus aureus, a 393 ug/ml solution cultures of Streptococcus faecalls, and a 161 ug/ml solution cultures of Streptocuccus mutans (see Table I).

Furthermore, on the average a 150 ug/ml and a 20 ug/ml solution also inactivaties the cultures of yeasts Candida albicans and Saccharomyces cerevisiae. A considerably lower concentration than the inactivation concentration ofthe drug preparation is useful as a growth inhibitor of the same organisms.

In addition, on the average a concentration of sanquinarine chloride presented in Table VI was sufficient per cubic centrmetre of media, to inhibit growth of some fungi known to belong to the group of ringworm-producing organisms.

TABLET Mean lnhibiting Dose of Sanguinarine Chlorine in M.icroorganisms ugiml of Media Microsporum can is 867 Microsporum nanum 650 Trich oph yton mentagrophytes 900 Trichophyton schoenleini 467 Trichophyton terrestre 467 Trich oph yton vanbreuseghemi 750 The basic preparations were, on numerous occasions, also tested on a variety of animals and also humans infected with ringworm and athletes foot fungus. It was found that preparations according to the present invention will considerably hasten the recovery from ringworm infection and facilitate rapid healing of lesions.

Results of clinical tests run by numerous dentists and detal institutions confirm that regular application of the composition of this invention to teeth surfaces will reduce the incidence of dental caries and the use on gums either by packing or irrigations will prevent or rapidly cure periodontal disease or mic robial infections of the gums and surrounding tissues.

Application of the compositions of the present invention to cold sores immensely hastens the heal ing process. Cold sores dry up and heal in a few days.

When animals suffering from scours are treated internally with preparations according to the present invention, the recovery rate exceeds that of the anti biotics commonly used for the purposes of treating scours.

Claims (30)

1. An antimicrobial composition comprising a mineral acid salt of a benzophenanthridine alkaloid and a non-toxic metal salt of an acid in a suitable solvent vehicle.

2. An anitmicrobial composition according to Claim 1, wherein the non-toxic metal salt is a fluoride, chloride, bromide, iodide, sulphate, nitrate, or acetate, or any mixture thereof.

3. An antimicrobial composition comprising a mineral acid salt of a benzophenanthridine alkaloid and a metal salt of an acid in a solvent selected from water, glycerine, propylene glycol, dimethyl sulphoxide, and C1-C6 alcohols.

4. A composition according to any of Claims 1 to 3, wherein the benzophenanthridine alkaloid is chelerythrine, sanguinarine, protopine, or homochelidonene, or any mixture thereof.

5. A composition according to Claim 4, wherein the alkaloid is sanguinarine.

6. A composition according to any of Claims 1 to 5, wherein the solvent is glycerine.

7. A composition according to any of Claims 1 to 6, wherein the mineral acid salt is sanguinarine chloride.

8. A composition according to Claims 1 to 7, wherein the metal salt is stannous fluoride or sodium fluoride.

9. A composition according to Claim 8, wherein the metal salt is stannous fluoride.

10. A composition according to any of Claims 1 to 7, wherein the metal salt is zinc chloride.

11. A composition according to any of Claims 1 to 4, wherein the alkaloid is chelerythrine.

12. A composition according to Claim 11, wherein the mineral acid salt is chelerythrine chloride.

13. A composition according to Claim 10, including 0.1-3% of a fluoride salt selected from stannous fluoride and sodium flouride.

14. An antimicrobial composition according to Claim 1, substantially as hereinbefore described with reference to any of the preparations and/or Examples.

15. A method for preparing an antimicrobial agent comprising: (a) dissolving a benzophenanthridine alkaloid in a mixture of chloroform and methanol; (b) acidifying the solution with a mineral acid to convert the alkaloid to an alkaloid salt; (c) evaporating the acidic solution to dryness; (d) recrystallizing the residue from a mixture of 50% ethanol and 50% chloroform; (e) dissolving the crystals in a solvent to make a solution of at least 0.3% by weight of crystals; and (f) mixing the solution with at least 35% by weight of a metal salt of an acid.

16. A method according to Claim 15, wherein the solvent is water, glycerine, propylene glycol, petrolatum, dimethyl sulphoxide, a C1-C6 alcohol, or any mixture thereof.

17. A method according to Claim 15 or 16, wherein the benzophenanthridine alkaloid is chelerythrine, sanguinarine, protopine, homochelidonene, or any mixture thereof.

18. A method according to Claim 17, wherein the alkaloid is chelerythrine.

19. A method according to Claim 17, wherein the alkaloid is sanguinarine.

20. A method according to Claim 15, substantially as hereinbefore described with reference to any ofthe preparations and/or Examples.

21. An antimicrobial agent prepared by a method according to any of Claims 15 to 20.

22. A method for preventing or treating infections in mammals, which comprises administering to the mammal an affective amount of the composition of any of Claims 1 to 14 and 21.

23. A method for treating ringworm in mammals comprising applying to the affected area an effective amount of the composition of any of Claims I to 14 and 21.

24. A method for treating scours in mammals comprising administering orally to an affected mammal an effective amount of the composition of any of Claims 1 to 14 and 21.

25. A method for treating periodontal disease comprising topically applying to the affected area an effective amount of the composition of any of Claims 1 to 14 and 21.

26. A method for treating dental caries comprising topically applying to the affected area an effective amount of the composition of any of Claims 1 to 14and21.

27. A method for treating skin tumours in mammals comprising topically applying to the affected area an effective amount of the composition of any of Claims 1 to 14 and 21.

28. A method for treating squamous cell carcinoma in mammals comprising topically applying to the affected area an effective amount of the composition of any of Claims 1 to 14 and 21.

29. A method for treating gingivitis and/or peridontitis comprising topically applying to the affected area an effective amount of the composition of any of Claims 1 to 14 and 21.

30. A method according to Claim 22, substantially as hereinbefore described with reference to any of the Examples.