NO872142L - SUBSTANCE, PHARMACEUTICAL OR COSMETIC MIXTURE AND PROCEDURES FOR PRODUCING THEREOF. - Google Patents

Abstract

Pharmaceutical or cosmetic substance or composition inhibiting or destroying at least one monocellular living being and/or at least one virus, medicament or product including such composition, process for producing such composition, chemical compound entering in such composition, and process for the inhibition or destruction of at least one unicellular living being and/or at least one virus. The inhibiting or destroying substance, according to the invention, is characterized in that it comprises at least one basic active principle inhibiting or destroying said unicellular living being and a principle for inhibiting or destroying at least one enzyme associated with said living being, forming an activator - particularly acting by synergy - of the basic active principle.

Description

The present invention relates to the inhibition or destruction of unicellular organisms such as protozoa, microbes, bacteria, gametes, fungi and viruses. The invention will therefore relate to technical areas such as contraceptives, antibiotic therapy, and wound inflammation in general either as a pharmaceutical or cosmetic product as well as agents for disinfection.

The term "inhibition" of unicellular organisms or viruses is understood to mean that one either prevents their reproduction, or makes it impossible for these organisms to perform certain functions that they normally perform. The term "destroy" means killing the single-celled organisms.

With respect to the present invention, the term means

"substance" any chemical compound or mixture of chemical compounds or association of such which has at least one given function or where a function is common to all the compounds,

and where said compounds can be added to the composition of the final product, usually associated with one or more diluents and possibly with other substances. The term "product" means a usable finished product. The finished product will thus usually consist of at least one diluent and several substances, where each substance may be composed of one or more chemical compounds with a similar or identical function. The term "substance" itself can be used to denote a real substance, but can also be purely theoretical and functional in the case of intricate mixtures where the compounds have several effects or where the effects influence each other. The above-mentioned classification in connection, substances and products will not necessarily correspond to the methods used for the production of the product and the mixture that is obtained in practice.

The term "mixture" is understood here and in the following

text a pharmaceutical or cosmetic substance.

A number of substances are already known which inhibit or

kill single-celled organisms, whether these substances are used purely biologically, pharmaceutically, cosmetically or as disinfectants.

Spermicidal compositions are thus described in U.S. Pat. the patents

4,339,441 and 4,359,475.

Numerous chemical compounds are also known which can be used in contraceptives. More specifically, surfactants are known which lower the interphase tension (e.g. quaternary ammonium compounds, nonoxynols, derivatives of amidoethyl glycinate of fatty acids, methyl tauridine sodium oxysalt) and other compounds such as phenylmercury nitrate, para-menthanyl-phenyl-polyoxyethylene ether or the trisodium salt of polysaccharide sulfuric acid ether which have inhibitory or killing effect on unicellular organisms, more particularly these compounds have spermicidal effects.

There are also known substances which mechanically act on spermatozoa, e.g. by making these immobile. U.S. patent 4

368 186 describes such effects and more particularly the mixture of "poloxames" and spermatozoa inhibiting agents, where the gel formation and dissolution of the products is controlled and thereby mechanically increases the effect of the inhibiting agents with an additive or synergistic effect.

Local anti-fertilization mixtures are also known,

especially spermicides, which include such chemical compounds, and which can be used locally to prevent fertilization itself, this applies to both humans and animals.

These known mixtures include a certain concentration of an inhibitory chemical compound and where the concentration lies above the minimal inhibitory concentration, hereinafter called the MIC, in a solution or suspended in a pharmaceutically acceptable diluent depending on the galenic form used. It is known that if you want to kill all the sperm that are present in 0.2 ml of semen in less than 5 sec. (these are the conditions specified in the total spermicidal test according to the standard of the IPPF, International Planned Parenthood Federation), then the concentration of the spermicidal active ingredient in 1 ml of the pharmaceutical mixture must be better than or equal to the minimum inhibitory concentration, i.e. MIC of the active ingredient. Said MIC depends on the type of compound involved, but also under which conditions the compound is used, i.e. in which galenic form is used in the mixture.

Pharmaceutical or cosmetic mixtures are also known which include at least one basic active ingredient which inhibits or kills at least one single-celled organism, or medicines or cosmetic products which include such mixtures.

In connection with wound treatment and disinfection, there are known numerous active ingredients such as halogens (chlorinated or iodinated derivatives), aldehydes, alcohols, phenols, acids, metals (silver, copper, mercury, zinc salts), amidines, biguanides, diphenylurea, oxidizing agents ( hydrogen peroxide, potassium permanganate), dyes, agents that lower the interphase tension and wetting agents (cationic, anionic, amphoteric or organic).

There are also known antiseptic and/or disinfectant and/or antiprotozoan and/or fungicide and/or antibiotic and/or antiviral products which include such inhibiting or killing substances in addition to methods for producing such substances and mixtures.

The known substances or compositions are usually satisfactory, but problems often arise in connection with their practical application.

These problems are essentially as follows:

Firstly, it is generally desirable to achieve a total inhibition or killing effect with regard to the single-celled organisms or for said virus, i.e. that in practice one wants 100% inhibition or killing. However, this is not always as easy without using very precise precautions or when a limit has been set without using very precise precautions or when a limit has been set with regard to the dose of the active ingredient in e.g. cosmetic products.

Furthermore, it is generally desirable to be able to have an activity against at least one or more given single-celled organisms or viruses, i.e. gametes or pathogens, without having short-term or long-term undesirable side effects (activity against other single-celled organisms, irritations, damage to the environment, etc.) . In practice, it has been found that the doses necessary to solve the above-mentioned problem often give rise to long-term side effects (usually associated with regular use), and even short-term side effects (this especially applies to the start of treatment). Purely medically, it has also been found that certain active ingredients have teratogenic and/or carcinogenic effects when certain dose levels are exceeded. When it comes to disinfection, unwanted effects can easily occur in the materials to be disinfected.

It is an aim of the present invention to increase the inhibiting or killing ability of a substance or mixture which includes a given limited concentration of an inhibiting or killing active ingredient without increasing this concentration. Furthermore, another purpose of the present invention is to reduce the concentration of the active ingredient without reducing the inhibitory or killing ability of the substance or mixture so that side effects can be avoided and the selectivity against selected unicellular organisms or viruses can be better regulated.

Furthermore, the invention provides new pharmaceutical or cosmetic mixtures, more particularly local contraceptives or products that include such mixtures, and which can either be used during attacks or when starting treatments, or regularly and continuously for long periods of time without risking side effects such as teratogenic, carcinogenic effects, and where the effect is total, i.e. that the percentage of errors is statistically equal to 0 or completely insignificant, and that this is within regular circumstances with regard to use (without any special precautions).

Furthermore, the invention provides a local anti-fertilization pharmaceutical product which can be easily used and which is totally effective, i.e. which gives the same results as those obtained when using oral anti-fertilization agents without having their disadvantages.

The invention also provides a chemical compound which can be used in a local contraceptive method.

Furthermore, it is a purpose of the present invention to reduce the doses of the active ingredients that inhibit or kill unicellular organisms or viruses in medicines, cosmetic products or disinfecting products, especially in antiseptic, anti-biotic, bactericidal, antiprotozoal, antifungal, spermicidal, antiviral products.

The present invention relates to a substance that inhibits or kills at least one type of single-celled organisms or viruses, especially protozoa, microbes, bacteria, gametes, fungi or others, in that the substance includes at least one basic active ingredient that inhibits or kills said single-celled organisms or viruses, and at least one component that inhibits or kills at least one enzyme associated with said living organism or virus, and where the latter component is an activator, preferably one with a synergistic effect on said basic active component.

The component that inhibits or kills at least one of the linked enzymes, hereinafter called "activating component" or "activator", is preferably an agent that prevents the function of the link between enzyme and substrate.

It has surprisingly been found that such an agent can be advantageously prepared using a fluoride anion F- which comes from a fluorinated compound either spontaneously or after a symatic effect. It has also been found that in certain cases the basic active ingredient is itself capable of developing an activating ingredient and/or that the activating ingredient is itself an active ingredient that inhibits or kills at least one unicellular living organism or virus.

More precisely, it has been found that such a fluorinated compound in itself has an ability to inhibit or kill unicellular organisms and viruses, and that such a fluorinated compound has a synergistic effect on the aforementioned basic active ingredient.

It is an aim of the present invention to provide a local anti-fertilization composition, more particularly a spermicidal composition characterized in that it includes at least one chemical compound capable of releasing or

develop a fluoride anion F-, as an active ingredient that inhibits or kills gametes, directly or by an amplification,

besides a diluent.

The invention also provides a medicinally active agent, an antiseptic and/or disinfectant, more particularly a topical antiseptic, and/or antiprotozoal agent, more particularly topical antiprotozoal agent and/or bactericide, more particularly topical bactericide, and/or antifungal, more particularly topical antifungal and/or antibiotic, more particularly topical antibiotic and/or antiviral product, a cosmetic product, more particularly a topical cosmetic product such as a synthetic soap or a milk, and an antifertility product that can be applied topically in contact with gametes, more especially in the form of creams, gels, solutions, foams, tablets, soluble products, tampons, vaginal suppositories, characterized in that they include a substance or a mixture according to the present invention.

The term "local" means that the product is used in a predetermined place to be processed, e.g. vagina or other anatomical parts, so that the effect is produced in the environment near this place and that it acts there against single-celled organisms or viruses that come into contact with the product, and that this treatment is in contrast to general delivery (e.g. oral delivery, or parenteral administration).

A substance, a mixture, a medicinal product, a common

product or a chemical compound according to the present invention concerns both local supply and general supply. Thus, a spermicidal product according to the present invention can be used locally. When, however, one wishes to avoid resistance effects that occur in pathogenic unicellular organisms in relation to regular therapy, then according to the present invention one will use general supply.

The invention also relates to the use in a cosmetic product or

in a mixture comprising at least one basic active ingredient and at least one fluorinated chemical compound capable of

releasing a fluoride anion F- as the active ingredient in said basic active ingredient; and with the use of a fluorinated chemical compound capable of emitting fluoride anion F- for the production of a substance comprising at least one basic active ingredient, as an activator for said basic active ingredient; besides the use of a substance or a mixture according to the present invention where one wishes to produce a medical product to be used as an antibiotic and/or antiprotozoal product and/or bactericidal product and/or antifungal product and/or antiseptic and/or antiviral product,

besides the use of substances or mixtures according to the present invention for the production of local contraception

agents both for animals and humans, more particularly for inhibiting or killing gametes.

The invention also concerns and provides manufacturing methods for substances or mixtures according to the present invention to inhibit or kill at least one single-celled organism, especially protozoa, microbes, bacteria, gametes, fungi or viruses, characterized in that at least one basic active ingredient that inhibits or kills said unicellular organism or virus is mixed with at least one component that inhibits or destroys the enzymatic system associated with said living organism or virus, and where the latter component is an activator for said basic active component.

The invention also provides a chemical compound comprising ionizable fluorine for use in a local anti-fertilization method for living beings, in particular a local anti-fertilization agent which can be used both for humans and animals according to the present invention, or in a substance or in a mixture according to the invention.

According to the present invention, an activating component which inhibits or kills or inactivates at least one enzyme which is associated with said single-celled organism or virus will induce a remarkable sensitization of said single-celled organism or virus against external influences, especially the effect of said basic active component which inhibits or kills said unicellular organism or virus, by "blocking" the linking enzyme and substrate. It is known that unicellular organisms or viruses have their own vital enzymatic systems. Furthermore, it is known that single-celled organisms or viruses can emit or secrete under certain circumstances (e.g. resistance to antibiotics) certain special enzymes which bring about the destruction of certain attacked (e.g. lactamase which destroys penicillins), and such enzymes are often called "protective enzymes". With the help of the present invention, the function of these enzymes will be inhibited, thus making the unicellular organisms more sensitive by creating optimal conditions with regard to the effect of the active ingredients. This means that one achieves far more advantages than what was previously known, on the one hand what is mentioned above, and on the other hand that one can selectively act on a given single-celled organism or obtain a given group of single-celled organisms by one acts on a given enzyme or on a group of enzymes. In connection with viruses, the present invention will inhibit the enzymes that are necessary for them to multiply or replicate. In what follows, the term "associated enzyme" will mean vital and/or protective enzymes found in unicellular organisms or in viruses.

It has been found that fluorine in its ionized state, F-, is particularly effective and has beneficial effects as an activating component against the associated enzymes, and this under irregular constellations. Furthermore, ionized fluoride F- is very common, economical and relatively easy to use. Its chemical properties and areas of application are easy to regulate, apart from the side effects that may arise in the constellations used according to the present method.

The present invention provides a method for inhibiting or killing at least one single-celled living organism or virus, more particularly protozoa, microbes, bacteria, gametes, fungi or others, characterized by using at least one basic active ingredient that inhibits or kills said single-celled living organisms and at least one ingredient that inhibits or destroys at least one enzyme that is associated with said living organism or virus, and where the latter ingredient is an activator, preferably one with a synergistic effect, for said former active ingredient, and that such a method includes using at least one substance or mixture according to the present invention.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to the inhibition or killing of unicellular organisms such as protozoa, microbes, bacteria, gametes, fungi, whether these are pathogenic or not, and viruses. The invention can be used in at least two different ways: either in connection with cosmetics or pharmaceutical products (in the following examples are given of spermicides, bactericides that act against pathogenic organisms), or can be used in more common ways, e.g. in agriculture, for disinfection or otherwise. In all these cases, a substance according to the present invention is characterized in that they include at least one basic active ingredient that inhibits or kills said single-celled organism or virus, and at least one ingredient that inhibits or kills or inactivates at least one enzyme associated with said living organism or virus , and where the latter component as such is an activator, preferably one with a synergistic effect, and which in itself is capable of inhibiting or killing said living organism or virus, on said basic active component.

The activating component according to the present invention functions in that it inactivates the enzymatic system associated with said living organism or virus. In this way, it has been found that such an activating ingredient is very advantageous as a means of preventing the coupling between an enzyme and substrate, and that one preferably uses a fluoride anion F", which is developed from a fluorinated compound, e.g. a metal derivative of fluorine. In particular, good results have been achieved with sodium fluoride, calcium fluoride, potassium fluoride, aluminum fluoride, stannous fluoride, ammonium fluoride or sodium monofluorophosphate or other compounds. You can, for example, use the present invention when treating or combating the following living organisms or viruses:

gametes (spermatozoa, egg)

gram-positive cocci such as staphylococci and streptococci gram-negative cocci such as gonococci

gram-positive bacilli

gram-negative bacilli such as colibacilli or escherichia

coli

acid-alcohol-fast bacilli such as mycobacteria, e.g.

mycobacterium smegmatis

spiral bacteria such as spirochetes, treponema various bacteria such as chlamydia

flagellate protozoa, such as trichomonas

different types of yeast such as candida albincans virus or retroviruses such as HIV (Human Immunodeficiency Virus) and herpes

For example, the following enzymes associated with at least one such unicellular living organism or virus can be inactivated by means of the present invention (Table 1):

A basic active ingredient that can be used in a substance or a mixture according to the present invention can e.g. be a cationic or anionic or amphoteric or nonionic surfactant that lowers the interphase tension, preferably a quaternary ammonium. Benzalkonium chloride or another alkyl benzalkonium is an example of a cationic surfactant. A methyltauride sodium oxysalt is an example of an anionic surfactant. An amidoethyl glycinate derivative of fatty acids is an example of an amphoteric surfactant. A nonoxynol is an example of a nonionic surfactant. The basic active ingredient can also advantageously be a phenylmercury nitrate or para-methanyl-phenyl-polyoxyethylene ether or a trisodium salt of a polysaccharide sulfuric acid ether or another compound (halogen, aldehyde, alcohol, phenyl, acid, metal, amidine, biguanide , diphenylurea, oxidising agent, dyes etc).

In various tests, it has been possible to show that the activating component alone is also directly active against the mentioned unicellular organisms or viruses. It has surprisingly been found that the connection between the activating ingredient and the basic active ingredient has a synergistic effect as the results obtained are not just the sum of the results that can be expected from the presence of the basic active ingredient alone or from the activating ingredient alone , but rather is better than this sum.

The effect of such active ingredients or activators can be measured by means of the minimally inhibited concentration called MIC, which corresponds to the concentration of the active ingredient that induces inhibition or death of all living organisms or viruses present within a given time.

In a substance according to the present invention, the anion-fluoride F- concentration developed by the activating fluorinated compound will advantageously be lower than the minimal inhibitory concentration (MIC) for the fluoride anion F- without the basic active ingredient.

Similarly, the concentration of the basic active ingredient in a substance according to the present invention will be less than the minimal inhibitory concentration MIC for this ingredient without the activating ingredient. Even then, and surprisingly, it has been shown that a substance according to the present invention has a total effect, i.e. an effect that is at least equal to that found for one of the substances, whether one only includes the fluoride anion F- in a concentration that is better or equal ingredient's MIC, or only a basic active ingredient at a concentration greater than or equal to its MIC.

Furthermore, the basic active ingredient itself may be able to develop an activating ingredient such as a fluoride anion F-. A substance or a mixture according to the present invention may include several basic active ingredients whose functions may be identical or different and/or different activating ingredients that act on at least one enzyme that is associated with one or more of the aforementioned basic active ingredients.

A substance or a composition or a mixture according to the present invention can be used in several different ways to obtain products, and as such products are used, these can be used for local or general supply.

Various therapeutic purposes can be mentioned as examples of particularly advantageous uses of products according to the present invention. The problem of inhibiting or killing single-celled organisms or viruses is an ever-increasing problem in therapy. This can sometimes be a fight against pathogenic organisms, and you can then use products that are antibiotic and/or antiprotozoal and/or bactericidal and/or fungicides and/or antiseptics and/or antiviral, besides that the products can be used against non-pathogenic organisms such as gametes, e.g. spermatozoa, and the products can thus be used as local contraceptives.

A substance or a mixture according to the present invention can advantageously also be used in a cosmetic product in such doses and concentrations that it is not classified as a pharmaceutical product.

The effective concentrations of the fluorinated chemical compounds and even of the basic active ingredient can be lowered to such a high degree that they lie below the threshold values that fill the pharmaceutical and cosmetic products, without this reducing the products' effectiveness.

A substance or a mixture according to the present invention can also be used in products that are neither cosmetics nor pharmaceutical products, e.g. as fungicides, antiprotozoal agents, antiseptics, antibiotics or as other products in connection with agricultural purposes, and also for disinfection of surfaces.

PREFERRED EMBODIMENTS OF THE INVENTION

The preferred embodiment of the present invention is a product that is locally applied to the genitalia of male and/or female mammals, e.g. such as spermicides and/or bactericides to combat sexually transmitted diseases (STDs).

Benzalkonium chloride and nonoxynol 9 are preferably used as active ingredients. As an activating component can use any metal derivative of fluorine, e.g. sodium fluoride. All known galenic forms can be used, preferably in the form of suppositories, creams, gels, solutions, foams, tablets, soluble products, tampons, vaginal suppositories or other forms.

The amounts to be used of the active and activating ingredient will depend on the galenic form, as only the actual concentration induced in vivo is important with regard to the product's effectiveness.

Said quantity ratios can vary between the MIC and the maximum concentration where you start to get side effects. In the case of local administration, irritation of the treated parts must be avoided. Usually, one also wants to avoid toxic concentrations.

The benzalkonium chloride concentration in vivo will thus preferably be 1.2% (per weight), while the fluoride anion F concentration in vivo is preferably 0.5% (per weight).

Several preferred embodiments of the invention will be described in the following and where different areas of application have been tried and where several different unicellular organisms have been used.

1) USE OF THE INVENTION FOR LOCAL FERTILIZATION OBSTRUCTION

The present invention provides a spermicidal pharmaceutical product which can be used locally so that it comes into contact with sperm and will thereby kill or inhibit the sperm cells present.

A spermicidal pharmaceutical product according to the present invention can be used in different galenic forms,

tablets, suppositories, soluble products, creams, gels, tampons, foams, vaginal suppositories that can be inserted into the vagina before sexual intercourse to prevent fertilization by killing or inhibiting the sperm cells before they come into contact with the egg cell.

A product according to the present invention consists of a composition which includes at least one chemical compound according to the present invention which includes ionizable fluorine, i.e. which is capable of emitting fluoride anion F- as an active spermicidal component, either directly or by amplification.

A chemical compound according to the present invention is

capable of emitting fluoride anion F~ when dissolved, preferably in an aqueous solution. The connection e.g. be a metal derivative of fluorine such as sodium fluoride, calcium fluoride, potassium fluoride, aluminum fluoride, stannous fluoride, ammonium fluoride, sodium monofluorophosphate or an organic fluorinated compound such as fluorinated amine.

When measuring the spermicidal activity of a chemical compound, you can use the total spermicidal sample specified according to the IPPF standard, where you find the minimal inhibitory concentration called MIC (expressed as a weight percent) of the chemical compound in 1 ml of solution which kills all sperm cells present in 0.2 ml of semen in a period of time that is shorter than 5 sec. The tests are performed on at least 6 sperm samples from different donors, using the following conditions specified by the IPPF:

trial age: two hours

density per mm3 : 50 million sperm cells

motility: 50% of the sperm should move rapidly forward when examined at 35 - 37°C in a new sample;

viscosity: ejaculate must appropriately be in liquid form,

and not stringy and must be homogeneous as can be observed with the naked eye;

it must be collected in sterile glass tubes that are sealed hermetically and stored at 37°C.

It has been found that under these conditions the fluoride anion F- will have a spermicidal activity of 100% according to the above-mentioned IPPF test when the anion is present in a concentration of 5 ppm (ie 5 mg/l).

A pharmaceutical spermicidal product according to a variant of the present invention includes at least one fluorinated chemical compound which is capable of emitting fluoride anion F- as the only active inhibitory or killing component. The concentration of F" in the composition or product should preferably be above 4.5 ppm, preferably above 5 ppm when a 100% inhibitory mixture or product is desired.

It has further been found that the fluoride anion F~, in addition to this direct spermicidal activity, also has a spermicidal activity in that it enhances known spermicidal compounds. A composition or mixture according to the present invention can therefore advantageously on the one hand consist of at least one basic active spermicidal component such as a cationic, anionic or amphoteric or non-ionic surface-active agent which lowers the interphase tension, or even para-methanylphenylpolyoskyethylene ether or a trisodium salt of polysaccharide sulfuric acid ether or another product, and on the other hand at least one fluorinated compound according to the present invention which is capable of emitting fluoride anion F- and finally a pharmaceutical diluent and various other known additives (antioxidants etc.).

In a composition according to the present invention, the concentration of the basic active ingredient can be below the MIC for this ingredient without the F ion, and the composition will nevertheless be 100% inhibitory, i.e. it satisfies the total spermicidal test according to the IPPF. A composition according to the present invention may also include a mixture of several basic active ingredients that inhibit gametes and/or contain a mixture of several different fluorinated chemical compounds.

All percentages in what follows are given by weight.

The preferred embodiments of the invention for use as local contraceptives are as follows:

SUPPOSITORIES.

Diluents: semi-synthetic glycerides or cocoa butter, or gelatin or glycerin and purified water, antioxidants and antiseptics.

CREAMS AND MILKS:

Diluents: distilled or purified water, wetting agents, emulsifiers, stabilisers, antioxidants, antiseptics (can be added in variable amounts depending on the viscosity and pH you want to achieve).

OINTMENTS AND POMADS:

Diluents: distilled or purified water, emulsifiers, thinners of the fatty compound type (vaseline, lanolein, lanovaselin, stearovaseline), stabilisers, antioxidants, antiseptics (can be added in variable amounts according to the desired viscosity and pH).

GEL:

Diluents: soluble derivatives of cellulose that are compatible with cationic surfactants, distilled or purified water, glycerin, sorbitol, antioxidants, antiseptics (can be added in variable proportions according to the desired viscosity and pH achieved).

SOLUBLE MATTER:

Diluents: polyvinyl alcohol, glycerin, sorbitol, propylene glycol, distilled or purified water, antioxidants.

PILLS:

Diluents: lactose, magnesium stearate, cellulose, amylum, citric acid and sodium bicarbonate.

SYNTHETIC SOAPS:

Diluents: foaming and wetting products compatible with quaternary ammonium ion (eg amphoteric surfactants such as betaine or amino-betaine), emollients, stabilizers, antioxidants and antiseptics.

SOLUTIONS:

Diluents: distilled or purified water, ethanol, antioxidants, glycerin, sorbitol, antiseptics (can be added in different proportions depending on the pH you want to achieve).

COMPARATIVE SAMPLE NO. 1

The MIC of various known spermicidal compounds was determined according to the IPPF test. The following results were obtained with 1 ml of composition reacting with 0.2 ml of sperm and determining the minimum concentration of the spermicidal compounds which killed all sperm within 5 sec. and where 6 sperm samples from different donors were used:

SAMPLE NO. 2.

The same total spermicidal test according to the IPPF was carried out with 1 ml of composition which already contained 0.0001% (by weight) of fluoride anion F- and where the test was carried out on 0.2 ml of semen by determining the minimum concentration of the spermicidal compounds which killed all sperm within 5 sec. and where 6 sperm samples from different donors were used.

It appears from the results that the inhibitory ability of known spermicidal components was greatly improved by adding ionized fluorine to the product. A product or composition according to the present invention may therefore contain very little of the basic active spermicidal component, more particularly the concentration may lie below the minimal concentration of this active component as it appears without F ions.

Furthermore, the concentration of F- ion in relation to a basic active ingredient can be very low, in particular it can be less than the minimally inhibitory concentration of fluoride anion F-.

It can e.g. it is mentioned that it has been found that when a solution contains 0.003% benzalkonium chloride and a solution containing 0.0001% fluoride anion F-, neither of these solutions has a total spermicidal activity according to the specified IPPF test. If, on the other hand, you have a solution containing 0.003% benzalkonium chloride and 0.0001% fluoride anion F", (here there must be a printing error, there must be three zeros after the comma), then this mixture will satisfy the IPPF test. One can thus observe a synergistic effect.

SAMPLE NO. 3:

This sample is identical to sample no. 1 and 2 above, but one could demonstrate that in the presence of sodium borate the ion F- is complexed, and the spermicidal effect of the F- ion disappears completely, which shows that only the F- ion is active or acts as an enhancing agent for the spermicidal effect.

SAMPLE NO. 4:

Under the same conditions as indicated in Samples 1 to 3, various perfluorinated benzalkonium chlorides were used as the basic spermicidal compounds. It could thus be shown that perfluorinated benzalkonium chlorides have a spermicidal activity that corresponds to that found for pure benzalkonium chloride. Furthermore, this activity becomes identical under the conditions indicated in sample no. 3 in the presence of sodium borate, this shows that the fluorine attached to the benzene nucleus is not ionized.

Test No. 2 was also performed when perfluorinated benzalkonium chloride was mixed with another chemical compound capable of developing F ion. The enhancement of the spermicidal activity of the perfluorinated benzalkonium chloride could then be observed. The test was also carried out in the presence of sodium borate (described in test no. 3), and this induced a spermicidal activity which was the same as that of perfluorinated benzalkonium chloride.

The two samples no. 3 and 4 showed that the strengthening of the basic active ingredient only takes place in the presence of the fluoride anion F-.

For the production of a spermicidal pharmaceutical mixture or a product according to the present invention, one will mix a solution with a given concentration of fluoride anion F- and a basic active ingredient in a diluent which is chosen according to the galenic form one wishes to produce. In connection with tablets, the basic active ingredient and the chemical compound that develops F ions will be mixed in a diluent which is then worked up in the form of a product.

COMPARATIVE SAMPLE NO. 5:

This in vitro test was performed on galenic preparations containing benzalkonium chloride as the basic active ingredient by determining the MIC of this compound after simulating in vitro the real conditions in vivo (extraction and solubilization).

This test, which was carried out without an activating ingredient, gave the following results:

The percentages correspond to the ratio (by weight) of benzalkonium chloride in the solution used in vitro for the spermicidal sample according to IPPF and obtained after simulation, the ratio being measured by titrating a sample taken from the solution.

SAMPLE NO. 6:

The same conditions were used as in comparative sample no. 5, but the galenic form originally contained 0.45% by weight of the fluoride anion F-.

The following results were obtained:

II) USE OF THE PRESENT INVENTION FOR THE TREATMENT OF ANTIBIOTICS, AS ANTIBIOTICS AND FOR THE COMBAT OF STD: The preferred embodiments of the invention for its use e.g. antiseptics in dermatology are the following:

CREAMS:

Diluents: distilled or purified water, wetting agents, emulsifiers, stabilisers, antioxidants, antiseptics (can be added in variable proportions according to the desired viscosity and pH).

OINTMENT:

Diluents: distilled or purified water, emulsifiers, fatty compound type diluents

(vaseline, lanolein, lanovaselin, stearovaseline)

stabilizers, antioxidants, antiseptics (can be added in variable amounts depending on the desired viscosity and pH).

SYNTHETIC SOAPS:

Thinners: foaming and wetting products compatible with quaternary ammonium compounds (e.g.

amphoteric surfactants such as betaine or amino-betaine), emollients, stabilizers, antioxidants and antiseptics.

SOLUTIONS:

Diluents: distilled or purified water, ethanol, antioxidants, glycerin, sorbitol, antiseptics (added in variable proportions according to the desired pH). Preferred embodiments of the invention for use as e.g. local antibiotics in dermatology are the following:

RESOLUTION:

Diluents: ethyl alcohol, propylene glycol, distilled water.

GEL:

Diluents: ethyl alcohol, hydroxypropyl cellulose, distilled water, glycerin.

CREAM:

Diluents: vaseline, vaseline oil, lanolin.

CREAMS:

Diluents: propylene glycol, polyoxyethylene glycol, distilled water. Preferred embodiments of the invention for use as local antibiotics in otorhinolaryngology are the following:

OPHTHALMIC CREAMS:

Diluents: vaseline, vaseline oil, lanolin

NASAL SOLUTIONS:

Diluents: distilled water, citric acid, sodium chloride.

To increase the activity of the local antibiotherapeutic preparations, it is advantageous to add 0.10% benzalkonium chloride. The preferred embodiments of the invention for use in connection with gynecology and more particularly for combating sexually transmitted diseases STD, are those described under the section "USE OF THE INVENTION FOR LOCAL FERTILIZATION HINDERANCE" and under the designation "SUPPOSITORY", "CREAMS", "OINTMENTS" ", "GELS",

"TABLETS", "SYNTHETIC SOAPS" and "SOLUTIONS".

The methods used in tests to elucidate these uses for the invention were as follows:

For Neisseria Gonorrhoeae:

The bacteria used came from the hospital isolation dilution area for the substances to be tested:

Solvent: bidistilled sterile water

Relationship with a geometric ratio of 2

Doses: from 2,000 to 1.56 (and less) ug/ml

2 ml of each dilution was mixed with 18 ml of solid culture medium: gelose medium for isolation of gonococci (Pasteur Institute) enriched with G supplement:

Culture medium for gonococci: 2,000 ml Pasteur medium +

500 ml G supplement.

These culture media and the indicated dilutions of the active substances were the tools used to investigate the bactericidal activity. The final concentration of the substances was from 800 to 0.156 ug/ml in the Petri dishes. The spent suspension (dilution in physiological water of a 24-hour culture of a nutrient-rich liquid medium) was placed in wells drilled in a synthetic plate. In each of the prepared wells, one breed was examined. Each dish was examined after 24 and 48 hours of cultivation (37°C).

Determination of MIC: The gonococcal strains were spread out on plates (dilution on plate) and some of them were prepared as comparative and control samples without the substances to be tested. 5 ul of 10 formed colonies/ml were deposited on gelose plates which partly and partly did not have the substances to be tested added (18 hour cultures of gonococci suspended in a Sørensen solution).

A 48 hour incubation at 36°C was carried out, after which the growth or lack of growth of the tested bacteria was observed.

For Candida Albicans:

The method is essentially the same as for Neisseria Gonorrhoeae with the following differences:

Dilution range in twice distilled water from 2,000

to 1.56 ug/ml.

0.5 ml of each dilution was added to 4.5 ml of liquid Roiron medium.

The tubes were supplemented with cultures of Candida (24 hours/Roiron medium) or with the comparative strain (Roiron medium). Incubation was carried out in an oven at 37°C for 24 and 48 hours. By

At t = 24 and 48 hours, a drop was examined under a microscope between a slide and coverslip.

The results are expressed by counting the yeast colonies and determining the fungicidal MIC. By comparing the tubes, the percentage resistance could be determined.

For Trichomonas Vaginalis:

Same procedure as for Candida Albicans.

For Chlamydia:

Inoculum: consist of host cells of chlamydia or Mac Coy cells. They were stored at -80°C. The cellular concentration was from 2 to 2.5 x 10 cells/ml for the individual samples. The cellular layer was prepared in a complete culture medium.

The substances to be tested were diluted in a geometric ratio of 2 in twice distilled water.

The method used is the one developed by the Professors

F. Catalan, P. Sednaoui, A. Milovanovic et al., A. Fournier Institute, Paris.

1 ml of the substance to be tested was mixed with 1 ml of cellular suspension (Mac Coy cells), and then incubated for 1

hour and 24 hours at 37°C. 0.2 ml of this mixture was then placed in the wells of the plates. The plates were then centrifuged for 1 hour at 2,000 rpm) and incubated for 1 hour at 37°C. The culture medium was then replaced with 0.2 ml of new medium containing 0.5 mg/ml cycloheximide, and the plates were incubated for 48 hours at 37°C. The cellular layer was then fixed with methanol and stained with a monoclonal antibody conjugated with FITC (fluorescein isothiocyanate). Inclusions that must be present in the Mac Coy cells were then observed under an inverted epifluorescence microscope.

The toxicity of the substances to be tested was therefore first investigated, and this was taken into account when the test was to be carried out on the host cells.

For Pseudominas Aeruginosa:

Preparation of a dense solution with two races.

Suspension in isotonic solution of NaCl (0.85% by weight)

It was checked that the suspension contained the same number of bacteria: standard technique in agel medium, using 1 µl of the successive dilutions of the original dense solution (geometric ratio 10). Incubation 18 hours at 37°C.

Preparation of dilutions of substances to be tested

in solution in twice distilled sterile water. Dilution ratio with a geometric ratio of 2 (eg from 2,000 to 0.015 ug/ml).

Mixture of Pseudominas Aeruginosa bacterial suspension (same origin as the AFNOR standard) with constant concentration (= 4 x 10<7> bacteria/ml) and with dilutions of the tested substances in decreasing order (2,000 to 0.015 ug/ml).

Incubation or contact time: 1 hour, 24 hours and 48 hours at 37°C.

The samples were run in parallel for each dilution of the substances. Observation of the number of survivors was compared with the corresponding average number for the comparative breeds that were grown at the same time as the samples.

Observation after 1 hour, 24 hours and 48 hours.

for Treponema, Gardnerella, Ducrey's bacilli, Streptococcus and Staphylococcus Aureus: The method used is described by professors F. Catalan, P. Sednaoui, A. Milovanovic, et al., A. Fournier Institute.

COMPARATIVE SAMPLE NO. 7

With benzalkonium chloride alone and nonoxynol 9 alone, the following results were obtained:

SAMPLE NO. 8:

The same test as described above was carried out in the presence of fluoride anion F-, the following results were obtained:

COMPARATIVE SAMPLE NO. 9 :

This test was carried out with substance containing benzalkonium chloride without the activating ingredient, but in the presence of serum proteins. The test gave the following results:

This sample demonstrates the known adverse effects of serum proteins on the activity of benzalkonium chloride.

SAMPLE NO. 10:

This test was carried out as indicated for test No. 9, but in the presence of the fluoride anion F- in the mixtures. The following results were obtained.

This test shows that the fluoride anion F- favorably reduces the unwanted effect of the serum proteins.

COMPARATIVE SAMPLE NO. 11:

This test was carried out with galenic forms in vitro (in the same way as tests No. 5 and 6), with products containing benzalkonium chloride as an active ingredient and no activating ingredient.

The following results were obtained:

The concentrations are those present in the fluid obtained after simulation and used in the sample.

SAMPLE NO. 12:

This test was carried out in the same way as test No. 11 with products containing 0.5% by weight of fluoride anion F-.

The following results were obtained:

III) USE OF THE INVENTION AS A DISINFECTANT:

This area of application concerns the treatment of floor surfaces, surfaces, instruments etc. that come into contact with bactericidal products.

The preferred embodiments of the invention for this purpose are as follows:

The tests were carried out according to standard AFNOR NFT 72-150, of March 1981 with benzalkonium chloride and then with nonoxynol 9 as active ingredient.

The neutralizing agent was as follows: 3% Tween 80 (V/V) and 0.3% lecithin (V/V). The pH of the medium was 7.2.

COMPARATIVE SAMPLE NO. 13:

This test was performed without an activating ingredient:

SAMPLE NO. 14:

This test is identical to the previous one but was carried out with fluoride anion F~ as activating component.

IV) USE OF THE INVENTION IN COSMETOLOGY:

The preferred embodiments of the invention with regard to cosmetology purposes are as follows: The following galenic forms can be used within cosmetology: creams, lubricating liquids, pomades, solutions, foam baths, synthetic soaps, shampoos, intimate articles and disinfecting liquids.

The composition with respect to diluents is the same as pharmaceutical preparations, but the concentration of benzalkonium chloride and anion F- are different.

These concentrations are preferably the following for all products: 0.2% benzalkonium chloride and 0.1% fluoride anion F-.

Chemical or natural active ingredients can also be included in these compositions and with the concentrations and doses that are permitted in cosmetology.

Creams and treatment liquids can be in a continuous aqueous phase (oil/water or water/oil emulsions) or as a paste when heated, or can be self-diluting in water.

Different types of diluents that can be used in different types of products are listed below: Humectants: glycerol, propylene glycol, diethylene glycol, glycol, sorbitol, polyoxyethylene glycol.

Emulsifiers: sodium stearate, beeswax, sorbitol ester, polyoxyethylene glycol ester, fatty alcohols, triethanolamine lanolin, tween, glycol stearate and polyglycols.

Stabilizers: glycol stearate, cetyl alginate alcohol, pectin, gum, fatty esters of polyols, soluble cellulose esters.

Antioxidants: tartaric acid, citric acid and ascorbic acid.

Antiseptics: Boric acid, benzoic acid, parabenzoic acid and their methyl or propyl esters with or without sodium ions.

PH: All these products are particularly effective when the pH is between 4.5 and 6.5. To achieve a pH within this range, citric acid will usually be used.

Claims (45)

1. Mixture to inhibit or kill at least one single-celled living organism or virus, especially protozoa, microbes, bacteria, gametes, fungi or others, characterized in that it comprises at least one basic active ingredient that inhibits or kills said single-celled living organism or virus, and at least one component that inhibits or destroys at least one enzyme associated with said living organism or virus, and where the latter component is an activator, preferably with a synergistic effect on the former basic active component. 2. Mixture according to claim 1, characterized in that the activating component is an agent that prevents the linking of enzyme and substrate. 3. Mixture according to any one of claims 1 to 2, characterized in that the activating ingredient is a fluorinated compound capable of emitting fluoride anion F-, preferably a metal derivative of fluorine such as sodium fluoride, calcium fluoride, potassium fluoride, aluminum fluoride, stannous fluoride, ammonium fluoride, sodium monofluorophosphate or another fluorine-containing compound. 4. Mixture according to claim 3, characterized in that the fluoride anion F- concentration emitted by the fluorinated compound is less than the minimum inhibitory concentration (MIC) of the fluoride anion F- without the basic active ingredient. 5. Mixture according to any one of claims 1 to 4, characterized in that the concentration of the basic active ingredient is less than the minimal inhibitory concentration MIC of this ingredient without the activating ingredient. 6. Mixture according to any one of claims 1 to 5, characterized in that the basic active ingredient is itself capable of emitting an activating ingredient. 7. Mixture according to any one of claims 1 to 6, characterized in that the activating ingredient is itself an active ingredient that inhibits or kills at least one type of unicellular living organism or virus. 8. Cosmetic mixture, characterized in that it contains in combination at least one basic active ingredient and at least one fluorinated chemical compound which is capable of emitting fluoride anion F- and which increases the effect of the basic active ingredient. 9. Local antifertility mixture, more particularly a spermicidal preparation, characterized in that it contains at least one chemical compound capable of emitting fluoride anion F~ as an active ingredient to thereby inhibit or kill gametes, either directly or by amplification, and a diluent. 10. Mixture according to claim 9, characterized in that the only active ingredient that inhibits or kills gametes is produced by at least one fluorinated chemical compound capable of emitting fluoride anion F-. 11. Mixture according to claim 10, characterized in that the concentration of F- in the preparation is higher than 4.5 ppm, preferably approx. 5 ppm to achieve a spermicidal activity of 100% according to the total spermicidal sample from the IPPF. 12. Mixture according to claim 9, characterized in that it contains at least one other compound as a basic active ingredient that inhibits or kills gametes and where its inhibitory or killing ability is increased in the presence of fluoride anion F- . 13. Mixture or preparation according to any one of claims 1 to 9 or 12, characterized in that the basic active ingredient is quaternary ammonium. 14. Mixture or preparation according to any one of claims 1 to 9, 12 or 13, characterized in that it contains a surface-active agent which can be cationic, preferably benzalkonium chloride, or another alkylbenzalkonium or anionic, preferably a methyltauridine sodium oxysalt; or amphoteric, preferably a derivative of amidoethyl glycinate of fatty acids or non-ionic, preferably a nonoxynol, and where said surfactant is a basic active ingredient. 15. Mixture or preparation according to each of claims 1 to 9, 12 to 14, characterized in that it contains phenylmercury nitrate or para-menthanylphenylpolyoxyethylene ether or a trisodium salt of a polysaccharide sulfuric acid ether as basic active ingredient. 16. Antiseptic and/or disinfectant, more particularly a local antiseptic product, characterized in that it contains a mixture or a preparation according to each of claims 1 to 15. 17. Anti-protozoal agent, more particularly a local anti-protozoal product, characterized in that it contains a mixture or a product according to any one of claims 1 to 15. 18. Bactericide, more particularly a local bactericidal product, characterized in that it contains a mixture or a preparation according to each of claims 1 to 15. 19. Fungicide, more particularly a local fungicide product, characterized in that it contains a mixture or preparation according to each of claims 1 to 15. 20. Antibiotics, especially a local antibiotic product, characterized in that it contains a mixture or a preparation according to each of claims 1 to 15. 21. Antiviral, more particularly a local antiviral product, characterized in that it contains a mixture or a preparation according to each of claims 1 to 15. 22. Cosmetic product, characterized in that it contains a mixture or a preparation according to each of claims 1 to 15. 23. Antifertility product that is used locally in contact with gametes, then especially in the form of suppositories, creams, gels, solutions, foams, tablets, soluble substances, tampons, vaginal suppositories, characterized in that it contains a mixture or a preparation according to any of claims 1 to 15. 24. Cosmetic product, characterized in that it contains at least one basic active ingredient and at least one fluorinated chemical compound which is capable of emitting fluoride anion F*, as an activator ingredient for said basic active ingredient. 25. Process for the production of a mixture or preparation, characterized in that a fluorinated chemical compound is used which is capable of emitting fluoride anion F- and also at least one basic active ingredient, and where the first-mentioned chemical compound acts as an activator for said basic active constituent. 26. Use of a mixture or preparation according to each of claims 1 to 15, for the production of a local fertility-preventing agent either for humans or animals, then especially for inhibiting or killing gametes. 27. Process for producing a mixture or a preparation that inhibits or kills at least one single-celled living organism, more particularly protozoa, microbes, bacteria, gametes, fungi or others besides viruses, characterized in that at least one basic active ingredient that inhibits or kills said unicellular living organism or virus, is mixed with at least one component that inhibits or inactivates an enzymatic system associated with said living organism or virus, and where the latter component is an activator for the former basic active component. 28. Method according to claim 27, characterized in that at least one activating component which inhibits or inactivates an enzymatic system is a fluorinated compound which is capable of emitting fluoride anion F-. 29. Use of a chemical compound that includes ionizable fluorine, in a local contraceptive method, more particularly in a local contraceptive product for humans and animals according to claim 23, or in a composition or preparation according to each of claims 1 to 15. 30. Use of a compound according to claim 29, in a preparation for inhibiting or killing gametes. 31. Compound according to each of claims 29 to 30, characterized in that it is produced using a metal derivative of fluorine such as sodium fluoride, calcium fluoride, potassium fluoride, aluminum fluoride, stannous fluoride, ammonium fluoride or sodium monofluorophosphate or another fluorine-containing compound. 32. A compound according to each of claims 29 to 30, characterized in that it is worked up using an organic fluorinated compound which is capable of emitting fluoride anion F", either spontaneously or after enzymatic action. 33. Method for inhibiting or killing at least one single-celled living organism or virus, especially protozoa, microbes, bacteria, gametes, fungi or others, characterized by using at least one basic active ingredient that inhibits or kills said single-celled living organism, and at least one component capable of inhibiting or inactivating at least one enzyme associated with said living organism or virus, and where the latter component is an activator, preferably with a synergistic effect on said active component. 34. Method according to claim 33, characterized in that an agent is used which prevents the coupling of enzyme and substrate. 35. Method according to each of claims 33 and 34, characterized in that at least one fluorinated chemical compound is used as an activating component which is capable of emitting fluoride anion F-, preferably a metal derivative of fluorine such as sodium fluoride, calcium fluoride, potassium fluoride, aluminum fluoride , stannous fluoride, ammonium fluoride or sodium monofluorophosphate or another fluorine-containing compound. 36. Method according to claim 35, characterized in that the concentration of the fluoride anion F- which must be released by the fluorinated compound is less than the minimally inhibitory concentration MIC for the fluoride anion F- without basic active ingredient. 37. Method according to each of claims 33 to 36, characterized in that the concentration of the basic active ingredient is less than the minimum inhibitory concentration (MIC) for this basic active ingredient without the presence of an activating ingredient. 38. Method according to each of claims 33 to 37, characterized in that the basic active ingredient used is one which is in itself capable of emitting an activating ingredient. 39. Method according to each of claims 33 to 38, characterized in that as activating ingredient one uses one which is in itself an active ingredient that inhibits or kills at least one type of unicellular living organism or virus. 40. Method according to claim 38, and according to each of claims 35 to 36, characterized in that the concentration of F~ in the preparation is above 4.5 ppm, preferably approx. 5 ppm, thereby achieving a spermicidal activity of 100% according to the total spermicidal sample from the IPPF. 41. Method according to each of claims 33 to 40, characterized in that quaternary ammonium is used as basic active ingredient. 42. A method according to each of claims 33 to 41, characterized in that a surface-active agent is used as basic active ingredient which can be cationic, preferably benzalkonium chloride or another alkylbenzalkonium compound, or anionic, preferably a methyl tauridine sodium oxysalt, or non- ionic, preferably a nonoxynol, or amphoteric, preferably a derivative of amidoethyl glycinate of fatty acids. 43. Method according to each of claims 33 to 42, characterized in that phenylmercury nitrate or para-methanylphenylpolyoxyethylene ether or a trisodium salt of a polysaccharide sulfuric acid ether is used as the basic active ingredient. 44. Method according to each of claims 33 to 43, characterized in that one uses a chemical compound which is capable of emitting ionized fluorine, preferably a metal derivative of fluorine or an organic fluorinated compound in a local method for preventing fertilization for living beings, more especially in a local contraceptive product for animals or humans. 45. Use of a chemical compound comprising ionizable fluorine, preferably a metal derivative of fluorine or an organic fluorinated compound, to inhibit or kill gametes.