FR2494117A1 - Dental health compsn. contg. biotin antagonist - to inhibit growth of biotin dependent cariogenic microorganisms - Google Patents

Abstract

Dental health prod. is a toothpaste, toothpowder, mouthwash, chewing gum, confection, tooth-coating concentrate or sustained-release buccal tablet contg. enough biotin antagonist (I) to prevent biotin uptake by microorganisms which cause dental caries, plaque and acid formation. Pref. (I) is a biotin inactivator and/or a biotin antimetabolite. (I) inhibit biotin which is an essential growth factor for the oral bacteria, so interfere with their development. They have no significant effect on the biotin store in the body.

Description

 The present invention relates, in general, to dental care products and relates in particular to products containing substances which neutralize the action of biotin, intended to halt the development and the effects of the microorganisms which produce dental caries in humans.

 In the current state of the art, no product is known which has the function of acting on the environment in which the oral bacteria live, so as to deprive them of the vitamins necessary for their development and which are normally present in saliva.

 The present invention is based on the idea of limiting to a very large extent the amount of biotin normally present in saliva and which is necessary for the development of microorganisms which are the main cause of the deterioration of teeth. It has been discovered that biotin is an essential substance for the development and life of these microorganisms and that the scarcity of biotin must considerably limit the number of these and, consequently, their possibility of causing dental caries. This scarcity of biotin is achieved by introducing various stomatological products neutralizing biotin into the mouth. An important factor which deserves to be noted is that antibiotic substances produce their effects directly in the mini-ecosystem which exists in the mouth; indeed, the biotin reserves of the body itself are not targeted and the importance of the antibiotic agents in question is such that it only results in an insignificant effect on the total biotin reserves of the patient's body.

 To make the invention understood, it is first necessary to take a look at the mechanism underlying the deterioration of teeth, the action of the vitamin "biotin", and the biotin requirements of iTticroorganisms which are at the origin of the deterioration of the teeth, as well as on the antibiotic substances which are able to block the normal ways by which these micro-organisms obtain and absorb this biotin.

 The mechanism by which dental caries appears appears to have two phases: first, the accumulation of plaques on the surface of the teeth by the action of a first type of microorganisms which produce hydrocarbon polymers such as dextrans. Second, the production of acids, usually by another group of microorganisms and it is these microorganisms that are the real cause of the destruction of the integrity of the dental structure.

 Acid producing microorganisms live in dental plaque; in addition to providing a protected environment for these bacteria, it can also provide them with food. In addition, the plaque can keep the acids produced in close proximity to the surface of the teeth, so these acids are less likely to dilute or even be washed off the surface of the teeth by other fluids in the mouth. .

 Dental plaque is mainly based on sucrose polymers and the microorganism which is most suspected as being responsible for its formation is Streptococcus mutans. This was reported by J.K. Clark in 192l (Brit. Jour.

Exp. Path. 5 141-147). A monumental work on this subject has been carried out more recently by J. Carlsson (Odont Revy. 18 55-74 (1967). Ibid 19 137-160 (1968). Ibid 19 Suppl. 14 (1968). In 1970 Carlsson sought to establish the food needs of the
Streptococcus mutans. Using a strain designated JC 2, which had been isolated from humans, he found that biotin was essential for growth (caries 4 305-320, 1970)
Acid-producing microorganisms have similar needs: in 244 Niven (Jour. Bact. 47 343-350) has shown that in the case of 21 strains of Streptococcus lactis examined to determine their vitamin requirements, all need biotin for growth. In 1950, SA Koser and
HJFisher have isolated 26 strains of Lactobacilli from human saliva and found that biotin was essential for the growth of all these strains (J. Dental Res. 29 700-773)
Although lactic acid can be considered as being the acid most often responsible for the deterioration of teeth (AT Brown in Present Knowledge of Nutrition, 1976, by the Nutrition Foundation, page 492) a participation in the demineralization of enamel and cement in superficial coronary and root caries, we cannot entirely exclude the possibility of a contributio of other acids, such as propionic acid. On this subject, EA Delwicke (J. Bact. 1949, 58 395 398) reports that out of 25 strains of
Propionibacterium, most need biotin for their growth.

 Biotin is a vitamin which intervenes in the respiratory function of living organisms and which seems necessary and almost all forms of life. Some organisms are able to synthesize their own biotin, while others obtain it from outside sources. Fortunately, many of the microorganisms that are involved in the production of dental caries require external sources of biotin. This suggests the possibility of creating a situation in which biotin has been rarefied in order to thus halt their development and this is clearly what the invention does. By following a diet which persistently blocks the absorption of biotin, can produce adverse conditions for both plaque and acid formation that are beneficial to dental health.

The depletion, or even complete removal, of the biotin necessary for the microorganisms that produce cavities is achieved by using stomatological products which introduce substances that neutralize biotin into the mouth. There are two categories of antibiotic substances. The first and most numerous are those called "bolite-biotinic antimeta
DW Woolley in his entitled book "A Study of Anti metabolites", Wiley 1952, defines horn antimetabolites as "artagonistic analogues for metabolites". The structural similarity with the active metabolites seems to be responsible for the binding of antimetabolites to sites normally intended for true metabolites; this blocks these sites and, at least temporarily, prevents true metabolites from attaching to the substrate. The coupling action with metabollets or with antimetabolites is considered to be a reversible reaction and lCs sites are considered to become free and can then be occupied for many times. The agent most likely to attaching to a free site depends strictly on the ratio of the concentrations of metabolites and antimetabolites in the environment considered; with equal concentra tios, an equal number of sites will be occupied by each of them. Thus, for only 1% of sites to be occupied by active metabolites, a 99/1 ratio between the concentrations of antimetabolites and metabolites is required.

 The literature lists a considerable number of compounds which are antimetabolites for biotin. However, not all modifications of the biotin molecule translate into antagonistic properties. For example, the compound known as "oxybiotin" in which un-oxygen has replaced the sulfur in the molecule, retains its vitamin activity in some cases.

In order to highlight the modifications made to the molecule to create antimetabolites, the structure of the initial active biotin is shown below

 It should be noted that the empty valences are occupied by hydrogens.

As examples of biotin antimetabolites, there may be mentioned
Desthiobiotin in which S has been replaced by 2H
Biotin sulfone with an S02 group replacing the S
Aliphatic ureylene-cyclohexyl acids where 2 C replace S and derivatives with shorter side chains.

 The biotin desthio-isQ in which S is replaced by 2 H and which has a geometric isomerism.

 The aliphatic sulfonic ureylene-tetra-ahydro-furyl acids with an oxygen replacing S and SO 3 H replacing COOL.

 Homouiotine with the addition of -CH2 - in the lateral cartilage.

 Ureylene cyclohexyl valeric acid in which a six carbon cycle replaces the four carbons and a sulfur the 5 element cycle.

 Alpha-dehydro-biotin with an alpha double bond with the COOH group.

 Alpha-methyl-biotin with a methyl group on the alpha carbon of the COOH group.

Alpha-methyl-desthiobiotin with an alpha methyl group linked to the COOH group and 2 H in place of S
Oxybiotin having a six-carbon side chain with an additional C in the side chain and replacing S with 0.

 Oxybiotin sulfonic acid with replacement of S by O and of COOH by SO3H.

The literature also lists
nor-biotin,
bis homobiotin,
tris homobiotin,
aliphatic 4 imidazole acids,
2 oxo-4 acid imidazolidine
thiazolidone
benzyl thioethers,
hydrazides and semicarbazides of biotin
bis hydrazides of suberic and seba acids
ciques,
1-3 methyl acetyl-4-thiazolidine carboxylate
1-2 propyl-3-acetyl-4-thiazolidine carboxylate
methyl ester and its hydrazide
gamma acid hydrazrazide (2 carboxy-3-i.dolyle)
butyric,
2-imidazolidone-4-carboxylic acid hydrazide
2-imidazolidone-4-caproic acid hydrazide and,
2-imidazolidone-4-valeric acid hydrazide.

 The choice of the biotin antimetabolite intended to be used in the present invention is not as difficult as the large number of possible antimetabolites might suggest. Indeed, certain compounds such as the antibiotic "thiazolidone" imply a risk of development of strains of bacteria resistant to antibiotics, which is obviously to be avoided. Other compounds, which are very different from those found in living organisms can be eliminated, in particular, considering the fact that other very similar ones are available. Those by making slight modifications to their molecule, such as the addition of a carbon in the side chain or an unsaturation achieved by removing an H from the side chain, would be promising as substances with low toxicity, but most logic is the use of desthiobiotin, because it has been definitively established that it is the precursor of biotin in certain microorganisms, especially in E. coli.

 There is a very high probability that the quantities of desthiobiotin which are ingested by humans can be used by the intestinal microflora for the production of biotin which, in turn, can be absorbed by the human body. It has been shown that a large part of the biotin supplied to the human body comes from its normal intestinal micro flora.

 The second category of biotin antagonists is that of substances which make biotin inactive, ie inactivating or "inactivating" agents which combine with biotin so as to render it incapable of fulfilling its functions of vitamins .

As examples of biotin inactivators, there may be mentioned
avidin found in the white of the eggs of birds and amphibians.

 the streptavidin produced by Streptomyces species.

A compound similar to avidin found in egg yolk. (Hw Meslar et al., J. Biol. Chem. V 253, n 19, pp 6979-6982 (1978), and also (JC McCuire et al. Biochem. J. 197 (2): 395-400, 1 Aug 1976)
A biotin-binding protein found in chicken plasma (RD Mandella @@ tal. Biochem @. 17 @ (2) 1978 p 629-634).

Avidin is the most abundant tax in this category and it is also the one known for the longest time and therefore the best known. It is a clycoprotein com place with a molecular weight of 6,000. It is present in chicken egg white in a proportion of 0.05%
Each avidin molecule has four active sites through which it can bind biotin. The link between avidin and biotin is very strong; the dissociation constant is 10 15 (NM Green. Nature, 217 (1968) p. 254). This combination effectively makes biotin unable to play the role of a vitamin.

 For the purposes of the invention, a member of choice from the category of biotin antagonists, rendering it inactive is avidin.

 Before deciding on the category of biotin antagonist that we want to adopt to rarefy the biotin available for oral microflora, it is important to examine the different effects of each of them. On the one hand, a biotinic inactivator such as avidin, when it is present in sufficient quantity to react with all the biotin present, will cause that no biotin is no longer available for the microflora. Since avidin has a molecular weight of 68,000 and biotin a molecular weight of 244.3 and one mole of avidin reacts with four moles of biotin, it can be seen that the stoichiometric ratio of the reactants is 68,000 / 4 x 24) 4.3 or 69.6 / 1. This result can be rounded to 70 parts by weight of avidin for one part by weight of biotin. Thus, as long as there is at least 70 times more avidin than biotin, the latter will be rendered inactive.

 With a biotin antimetabolite present in the proportion of 70 parts of antimetabolites for one part of biotin, the activity of the latter will be reduced to 1 / 70th of its normal value, which is undoubtedly a valuable ap result, although not constituting not a complete reduction to almost zero.

 When the concentration of the biotin antagonist decreases, for example, between applications eg @ as long as biotin continues to be provided by saliva, the relative effectiveness of the biotin antagonist, while being) resistant, changes according to its nature . With avidin, for example, halving, that is, 35 parts of avidin for one part of biotin, can only make half of the biotin inactive. A reduction of the antimetabolite from 70 parts to 35 parts for one part of biotin results in a variation of the activity of biotin from 1 / 70th to 1 / 35th.

Therefore, biotin antimetabolite has the advantage when the more favorable ratios begin to dlm, inuer.

In the following examples, the calculations on the quantities of biotinic antagonists to be incorporated are based on data found in the manual "The Biochemists Habdbook, Cyril Long, Editor, published by Van IJostranden 1961. We find in this following significant data
average saliva flow at "rest": 30 ml / h limits: 2.5 to 110 ml / h
average saliva flow under stimulation: 114 ml / h limits: 24 to 288 ml / h
Biotin content of saliva: about 0.8 micrograms per liter.

To ensure efficacy in all cases, the following calculations of the biotin antagonist concentrations are all based on the maximum saliva flow rate, and therefore on the maximum release of biotin in the mouth. It is the constant replacement of biotin in the oral cavity by saliva which must be countered by the treatment envisaged by the invention and to be really effective a tireless effort must be made to prevent the microflora from accessing biotin which is essential to him. To make this more possible, the invention provides various means for introducing biotinic antagonists into the oral cavity.
toothpaste,
mouth rinses, i.e. mouthpieces,
chewing gum,
confectionery,
dental coating concentrates and,
the late-release tablets ensuring a pro release
length of active agents.

We will find below some examples of formulas intended to illustrate these various means
TOOTHPASTE
The toothpastes according to the invention can be presented either in the form of paste or in the form of powder.

 Toothpaste contains the usual substances such as water, abrasives, humectants, surfactants, perfumes and they can also contain compounds providing fluoride ions or stannous ions and other substances which occur therein - Even in the minds of experienced technicians, but more particularly and specifically, they contain biotinic antagonists. In addition, since certain usable components such as with dine can be subject to microbial attack, it is useful to incorporate an antiseptic or a combination of antiseptics, such as methyl- and propyl-parahydroxylbenzoates.

 The ratio of biotin antagonist concentrations used is as follows: the amount of saliva in the mouth, in the absence of stimulation, is remarkably low, less than 10 ml and often only around 4 to 6 ml. With a biotin content of 0.8 micrograms per liter of saliva, this amounts to saying that there should be no more than .008 micrograms of biotin inactive during toothbrushing. By adopting a ratio of 70/1 for the biotin antagonist, this corresponds to 0.56 micrograms of biotin antagonist per operation. A "Ribbon" of toothpaste, like the one usually applied to the toothbrush, weighs around one gram to one and a half grams.

To have a certain margin of safety, we suppose the use of the lower quantity, that is to say of one gram.

In other words, each gram of toothpaste should provide 0.56 micrograms of biotin antagonist, or 56 micrograms per 100 grams.

 Since the exposure to the biotin antagonist during brushing of the teeth is only short-lived, and the surfaces to be treated are not all easily accessible, the Applicant prefers to increase the concentration of biotin antagonist to ensure efficiency, that is, use 560 micrograms per 100 grams of toothpaste.

EXAMPLE 1
Toothpaste containing an antagonist-type biotin inactivating agent.

Calcium pyrophosphate 40 X
Glycerol 20%
Sorbitol 10%
Sodium lauryl sulfate 1.5
Carboxy-methyl-cellulose 1%
0.2% methyl parahydroxy benzoate
Saccharin 0.12%
0.05% propyl parahydroxy benzoate
Avidin 0.00056%
Perfume according to taste
Water qs 100
There is adsorption of avidin on bentonite, this adsorption represents a first step of a known means for isolating avidin from egg white (H. Fraenkel-Conrat et al.) It is practical to incorporate the avidin in compositions of the invention in the form of an adsorbate on bentonite.

In the cited reference, the avidin bentonite ratio in the adsorbate is 1/250
EXAMPLE 2
Toothpaste containing an avidinbentonite adsorbate as an antagonist-type biotin inactivating agent
Calcium pyrophosphate 40%
Glycerol 20.0
Sorbitol 10%
Sodium lauryl sulfate 1.5%
Carboxy-methyl-cellulose 1 #
Adsorbate 1/250 avidin bentonite 0.14%
11
0.2% methyl parahydroxy benxoate, accharir.ej
0.05% propyl parahydroxy-benzoat @
Perfume according to taste,
Water qs 100
EXAMPLE 3
Toothpaste containing a biotin antimetabolite read antagonist type.

Calcium pyrophosphate 20 /
Giycerol 20%
Sorbitol 10%
Sodium lauryl sulfate 1.5%
Carboxy-methyl-cellulose 1%
0.2% methyl parahydroxy benzoate
Saccharin 0.12%
Propyl parahydroxy benzoate C, 005%
Desthiobiotin 0,00056%
Perfume according to taste
Water qs 100
EXAMPLE 4
Tooth paste containing an antimetabolite of biotin and an inactivator of this one.

Calcium pyrophosphate 40%
Glycerol 20%
Sorbitol 10%
sodium lauryl sulfate
Carboxy-methyl-cellulose 1%
0.2% methyl parahydroxy benzoate
Saccharin 0.12 7
0.05% propyl parahydroxy benzoate
Avidin 0.00028%
Desthiobiotin 0,00028 7
Perfume according to taste
Water qs 100
EXAMPLE 5
Stannous fluoride toothpaste containing alpha-methyl-biotin as antimetabolite thereof antagonist type
Calcium pyrophosphate 4)%
Glycerol 20%
Sorbitol 10%
Sodium lauryl sulfate 1.5%
Carboxy-methyl-cellulose 1%
Stannous fluoride
Methyl parahydroxy benzoate 0.9%
Saccharin 0.12%
Parahydroxy-benzoate dp propyle 0.05 d
a-methyl biotin G, 00056%
Perfume according to taste
Water qs 100
EXAMPLE 6
Tooth powder containing an adorbate 1/250 of vadine and bentonite as deactivating antagonistic biotin.

Calcium pyrophosphate 52%
Sorbitol 40%
Mannitol 2%
Sodium lauryl sulfate 2%
Carboxy-methyl-cellulose 1.43%
Avidin-bentonite adsorbate 1/250 0.3%
0.40% stannous fluoride
Methyl parahydroxy benzoate 0.20%
Saccharin, 12
0.05% propyl parahydroxy benzoate
Spray-dried perfume 1.50%
The previous and following toothpaste formulas have been calculated based on the following considerations: when using a toothpaste, the "dose" applied to the toothbrush is slightly less than the amount of toothpaste normally used. believes that each dose is close to half a gram; since the latter can deviate to a certain extent from this figure, the Applicant uses a ten times greater proportion of biotin antagonist in order to ensure efficacy.

 since the quantity of toothpaste is only half that used in the form of a paste, the proportion of the biotin antagonist has been doubled for the powder in the formulas which follow.

EXAMPLE 7
Tooth powder containing an antagonistic biotin antimetabolite.

Calcium pyrophosphate 52%
Sorbitol h0%
Mannitol 2.4789%
Sodium lauryl sulfate 2%
Carboxy-methyl-cellulose 1.43%
Stannous fluoride 0.80%
Methyl parahydroxy-benzoate 0.20%
Saccharin 0.12%
0.05% propyl parahydroxy benzoate
Desthiobiotin 0.0011%
Spray-dried perfume 1.50%
EXAMPLE 8
Tooth powder containing biotin antagonists of both the inactivating type and the antimetabolite type.

Calcium pyrophosphate 52%
Sorbitol 40%
Mannitol 2.4789%
Sodium lauryl sulfate 2%
Carboxy-methyl-cellulose 1.43%
Stannous fluoride 0.80
Methyl parahydroxy-benzoate 0.20%
Saccharin 0.12%
0.05% propyl parahydroxy benzoate
Avidin 0.0006%
Desthiobiotin 0.0005%
Spray-dried perfume 1.50%
The mouthwashes prepared according to the invention have a different purpose from most of those currently offered. While the latter seem to have the objective of practically eliminating the microflora from the oral cavity, their action is, in reality, to a large extent aesthetic measure. On the other hand, the use of mouthwashes according to the invention must be considered as one of the multiple means brought by it to preserve the integrity of the teeth and; this with maximum efficiency.

 The calculations for determining the proportion of active substances to be incorporated into mouthwashes are similar to those used for toothpaste and pastes, the determining factor being that it is to oppose the action of biotin in the mouth at the time of use. As a basis for calculation, it is again considered that a quantity of 10 ml of saliva is more than one normally meets and one applies a tenfold dose of biotin antagonist. The quantity of mouthwash most often used each time is between 15 and 30 ml. Using the smallest number increases the assurance of effectiveness. As in the case of toothpaste, the dose of biotin antagonist required per use is approximately 0.56 micrograms by rounding this figure and compensating the slight decrease in density due to the presence of alcohol in the solution and by multiplying by a safety factor of 105 we obtain a figure of the order of 40 micrograms of biotin antagonist per 100 grams of mouthwash.

EXAMPLE 9
Mouthwash containing an inactivating biotin antagonist.

Alcohol SD 38 B 12% by weight
Glycerol 10%
Sodium lauryl sulfate 2%
Polysorbate 20 1% "
Carboxy-methyl-cellulose 0.5% "
0.2 "Methyl parahydroxy benzoate
0.05% propyl parahydroxy benzoate "
Sodium saccharin 0.02%
Avidin 0.00004
Perfume according to taste
Water qs 100
EXAMPLE 10
Mouthwash containing a biotin antagonist of the antimetabolite type.

Alcohol SD 38 B 12% by weight
Glycerol 10% by weight
Sodium lauryl sulfate 2% "
Polyscrbate 20 1%
Carboxy-methyl-celulose 0.5%
Methyl parahydroxy benzoate Q, 2%
Propyl parahydroxy benzoate 0.005%
Sodium saccharin 0.02%
Desthiobiotin 0.00004% I '
Perfume. according to taste
Water qs 100 EXAMPLE 11.

 Mouthwash containing biotin antagonists of both the deactivating type and the antimetabolite type.

Alcohol SD 38 B 12% by weight
Glycerol 10%
Sodium lauryl sulfate 2%
Polysorbate 20 1%
Carboxy-methyl-cellulose 0.5%
0.2% methyl parahydroxy benzoate "
0.05% propyl parahydroxy benzoate "
Sodium saccharin 0.02%
Avidin 0.00002%
a-dehydrobiotin 0.00002%
Perfume according to taste
Water qs 100
Any means by which biotin antagonists can be introduced into the oral cavity is considered to be advantageous, as explained above; all those which make it possible to maintain sensitive levels of these active agents in the mouth are even more advantageous. Chewing gum or "chewing gum" have the characteristics placing it in this category.

Chewing gum has the power to extend the release time of its components; this is universally known from experience regarding the release of fragrances and sweeteners. Even before they are actually released, the biotin antagonists present in the gum matrix can come into contact with biotin and can react with it
The relative calculations: .x optimal proportions of biotin tagonists to be incorporated into the chewing gum took into account the factor which has not been emphasized before and which is the time.

Biotin is introduced into the syrup through saliva. The inflow of saliva is continuous and crnrne it was my very highest, we measured a maximum flow rate of 288 ml / h under certain stimulation conditions, which should correspond to the conditions that exist while chewing the rubber . The
The Applicant used a figure of 30 ml / h for the calculations and based on a period of a lure during which a piece of gum was chewed. After this period of time, the scent of the gum is practically exhausted and it is ready to be thrown in. With a biotin content in saliva of 0.8 microgram / liter, the 300 ml produced in the space of one hour brought 0.24 micrograms of biotin.

 With a biotin-to-biotin antagonist ratio of 70/1 this requires 16.8 micrograms of antagonist. Thus, at the beginning of the period of time considered, there will be a significant excess of biotin antagonist, while the situation towards the end of the hour is relatively uncertain, this is the reason why the Applicant prefers to incorporate practically double the calculated concentration, i.e. 30 micrograms per piece of gum.

 There are certain aspects of the situation which can be considered favorable after the gum has been chewed for some time. On the one hand, the storyteller sugar in the gum dissolved and was swallowed. In addition, the continuous washing action which occurs from this point has the effect of reducing the concentration of residues of food materials which can ordinarily serve as food for bacteria; the combination of reducing the amount of biotin available and that of food can only be beneficial to the user.

 A precaution which must be observed during the preparation of the product when using avidin as a biotin antagonist concerns the temperatures to which this niece is exposed. Although avidin is slightly less sensitive to heat denaturation than the egg white itself, it should not however be exposed to temperatures above 900 C and, preferably, should not be incorporated in the melted gum before it has cooled below 600 C. When avidin is the agent used, it may be advantageous to add it in the form of the avidin / bentonite adsorbate previously described, because bentonite is frequently used in chewing gum.

EXAMPLE 12
Chewing gum or "chewing gum" containing as an antagonist of biotin an agent of inactivation of this one.

Chewing gum, 3 gram blister
Avidin 30 micrograms
EXAMPLE 13
Chewing gum containing an antimetabolite as biotin antagonist.

Chewing gum, 3 gram blister
Desthiobiotin 30 micrograms
EXAMPLE 14
Chewing gum containing biotin antagonists of both the inactivating and antimetabolite types.

Chewing gum, 3 gram blister
Avidin 15 micrograms
a-methyl-desthiobiotin 15 micrograms
Another form of vehicle to introduce biotin antagonists into the mouth is confectionery, and especially hard candy-like candy, which takes some time to dissolve in the mouth. protection of dental health targeted by the invention.

 Using a confectionery having the dimensions of the well-known candies called "Life Savers" as a model, the Applicant has found that each piece, weighing around 2.5 grams, takes approximately 10 to 1 minutes to dissolve in the mouth. This last figure is the one that is accompanied by a larger amount of biotin to be released with saliva, and it is this value that served as the basis for the following calculations. 12 minutes representing the 5th of an hour and with a maximum flow of saliva under stimulation of 00 ml / h, 50 ml of saliva are released in this period of time. For 0.8 micrograms of biotin / liter, the 60 ml of saliva releases 0.048 micrograms of biotin, a figure which can be rounded up to 0.05 micrograms.

 These 0.05 micrograms of biotin would require 3.5 micrograms of antagonist according to the principles of the invention, in order to oppose a possible use of this vitamin by the microflora. The Applicant prefers to incorporate double this amount in order to be more sure of the effectiveness of the product. This is equivalent to using 7 micrograms of biotin antagonist in each 2.5 gram piece of hard candy that takes 12 minutes to eat. Thus the percentage rises to 280 micrograms.

 Since there are so many types of confectionery, it may be useful to base calculations on the amount of biotin antagonists to be added based on the time normally required to ingest a piece of the particular confection being considered . In the previous paragraph, it was shown that a dissolution time of 12 minutes corresponded to a dosage of 7 micrograms of biotin antagonist; this corresponds to 0.58 micrograms of antagonist / minute of the time required to dissolve or ingest a particular type of confectionery. The term "ingested" is probably the most appropriate, in particular, in the case of chewy confectionery.

 As indicated in the previous paragraph, concerning chewing gum, when there is a risk that the avidin constituting the biotin inactivating agent is exposed to high temperatures, certain precautions must be taken to avoid incorporating this avidin at temperatures approaching 900 ° C .; this addition should preferably be delayed until the molten candy has cooled to at least 600 ° C in order to avoid any risk.

 Since the biotin antagonists used in confectionery are called to be ingested, the Applicant prefers to limit the representatives of the two categories of biotin antagonists to only two, in particular, to avidin for the inactivation agents and to desthiobiotin for antimetabolitic agents. Avidin is a natural component of eggs, which has been ingested by acme for many centuries; desthichiotin, as mentioned above, was; found capable of acting as a precursor to the biological synthesis of biotin.

 In order to include other types of confectionery as examples, tests were carried out to determine the average consumption times of caramels weighing 9 grams each and also of chocolate confectionery bars weighing 5 grams each. For the former, the average consumption time is close to 90 seconds, while for the latter, it spans a relatively wide range between approximately 3 and 4 minutes, so that the figure of 4 has been used here. minutes.

 The caramels require, under these conditions, 0.87 micrograms of biotin antagonist over 9 grams, while the candy bars require 2.32 mierograms per bar of 58 grams.

EXAMPLE 15
Hard candy type confectionery containing itavidin as an antagonist biotin inactivating agent.

Hard candy base 100 grams
Avidin 280 micrograms
EXAMPLE 16
Caramel-type confectionery containing desthiobiotin as a biotin antagonist of the antimetabolite type.

Caramel base 100 grams
Desthiobiotin 10 micrograms
EXAMPLE 17
Chocolate confectionery bar containing inactivating type and antimetabolite type biotin antagonists.

Confectionery bar base 100 grams
Desthiobiotin 2 micrograms
Avideine 2 micrograms
A comment concerning the addition of avidin to confectionery is essential at this point; since the cakes are edible, it is perfectly possible to add avidin, when it is desired to include @eileci, by adding appropriate quantities of egg white. To calculate the quantities of egg white to be added, it suffices to recall that, when dry, the egg white contains almost 0.05% avidin.

 The fact that the use of oiotin antagonists, as recommended in this specification, is safe, will be demonstrated in a later paragraph, and it is believed that it will also show that it does not entails no risk with regard to the supply of biotin to the body by normal routes. However, in the case.

where someone would like to further guarantee that by ingesting abnormally large amounts of confectionery containing biotin antagonists, their biotin supply would not be diminished, it is possible to produce confectionery in which biotin is supplied in amounts equivalent to those which can be canceled by the antagonists present in an outer layer.

 Hard candies lend themselves best to this embodiment. After the body or core of the confectionery containing the biotin antagonist has been prepared, the coating containing the biotin is added. This can be accomplished by double tableting or by coating.

 To calculate the concentrations to be incorporated in the confectionery, we first determine the dissolution time of the nucleus of a piece of candy, we calculate the dosage of biotin antagonist using the factor of 0.56 micrograms per minute of time of dissolution, and the dosage of the biotin to be incorporated in the coating of each part is fixed at 1/7 of the amount of antagonists.

 The number of types of candy that have been imagined is almost innumerable; above we have considered a type of candy which is not intended to be chewed. The use of the invention can be extended to a large number of others intended for chewing by advising the user to hold the candy in the mouth for a few moments, until the coating containing the Biotin has dissolved. A practical device to point out this point could consist in using a different perfume in the coating and in the core of the confectionery, so when the user feels the change in flavor or taste, he is informed that the coating has dissolved and the biotin has been swallowed.After that, he can chew the confectionery with the assurance that the benefits of the invention are preserved.

EXAMPLE 18
Hard candy with a coating containing biotin and in the nucleus an inactivating type biotin antagonist.

Hard candy bar 100 grams
Avidin 280 micrograms
After the pellets have been formed, they are placed in a coating machine and the following composition is applied to them
Sugar 5 grams
Water 5 grams
Biotin 4 mircograms
EXAMPLE 19
Hard candy with a coating containing biotin and the core of which contains a biotin antagonist of the antimetabolite type.

Hard candy base 100 grams
Destiiiobiotin 280 micrograms
After the pellets have been formed, they are placed in a coating machine and the following solution is applied to them.
Sugar 5 grams
Water 5 grams
Biotin 4 micrograms
EXAMPLE 20
Hard candy comprising a coating containing bictin and the core of which contains biotin antagonists of both the antimetabolite type and the inactivating type.

Hard candy base 100 grams
Desthiobiotin 140 micrograms
Avidin 140 micrograms
After the pellets have been formed, they are placed in a coating machine and the following solution is applied
Sugar 5 gram
Water 5 grams
Biotin 4 micrograms
Another means for applying the principles of the invention is represented by the dental coating concentrates, the description of which follows. It is a solution intended to be applied to the teeth between meals and which contains a significantly higher concentration of biotin antagonist than mouthwashes. It also differs from the latter in that it is not intended to be rinsed after use. Preferably, the concentrate is prepared to have as mild a taste as possible so as not to stimulate the production of saliva since this would also have the effect of bringing additional biotin to the scene. The concentrate is intended to remain on the surface of the teeth or its biotin antagonist content may be the most effective in blocking the adsorption of biotin by microorganisms producing dental caries,
The content of the biotin antagonist in the concentrate is based on the length of the period of time that is expected to remain effective. The calculations that follow are based on an hour's duration. Longer periods are affected by increasing uncertainty that results from effects such as the subject swallowing, speaking, smoking, or doing some other activity that may have the effect of displacing the coating of the teeth.

 In the absence of stimulation, the maximum production of saliva is indicated in the reference cited above as being 110 ml / h. Consequently, the quantity of bictin present in these 110 ml is therefore the maximum which must be rendered inoperative by a single treatment and at a concentration of 0.8 micrograms per liter of saliva, this corresponding to 0.088 micrograms of biotin. In the 70/1 biotin / antagonist ratio, this requires a dosage of 6.16 micrograms of the latter. Thus, 10 micrograms corresponds to a certain excess and constitutes a practical factor for the calculations.

Using either a tampon or a sprayer, 1 ml is ideal for producing a
2 coating on the teeth. With an @osure of 10 micrograms / 1/2 @@@, the percentage of biotin antagonist in the concentrate amounts to 2000 micrograms or even more simply to 2 milligrams. To avoid the danger that a porsonne will take excessive amounts of this concentrate, it is wise to package it in 5 ml bottles, with restricted openings, a bit like the product @ onn @ under the name "Binaca" qul is sold commercially to freshen the breath.

 11 es. It is reasonable to expect that the surfaces of the teeth which come into contact with the tongue will be freed from the concentrate soon after its application. This is not really a problem because very few are the cavities that develop on surfaces. The critical zones are those which are sheltered: the gaps between the teeth, at the level of the gums, in other words, the locations or the deposits remain physically more or less "hidden" and undisturbed.

EXAMPLE 21
Concentrate of dental coating containing an antimetabolite type biotin antagonist.

Glycerin 30%
Carboxy-methyl-cellulose 2%
Bentonite 27.

0.2% methyl parahydroxy benzoate
0.05% propyl parahydroxy benzoate
Desthiobiotin 0.002%
Perfume according to taste
Water qs 100
The avidin separation adsorbate intended to purify the latter.

EXAMPLE 22
A dental coating concentrate containing an inactivating biotin antagonist.

glycerin 30%
Carboxy-methyl-luiose 2%
1/250 avidin bentonite adsorbate 0.5%
Rentonite 1.5%
0.2% methyl parahydroxy benzonte
0.05% propyel parahydroxy benzoate
Perfume according to taste
Water qs 100
Other substances which could be incorporated in this concentrate will present themselves to the mind of the informed technician, such as for example surfactants, agents for adjusting viscosity and additives intended to improve the aesthetic appearance. , without departing from the scope of the invention. The proportions of biotin antagonists could also be changed to produce an action.

for different periods of time or to vary the excess quantities. Other biotin antagonists could also be incorporated, as well as mixtures of these could be used.

 The following example illustrates a formula that uses a mixture of biotin antagonists.

EXAMPLE 23
Concentrate of dental coating containing biotin antagonists of both the anti-metabolite type and the inactivating type.

Glycerin 30%
Carboxy-methyl-cellulose 2%
bentonite 2%
0.2 S methyl parahydroxybenxoate
Propyl parahydroxy benzoate 0.005%
Desthiobiotin 0.001%
Avidin 0.001%
Water qs 100
Another means, by means of which the microflora of the mouth can be deprived of the biotin which it needs is represented by long-lasting sucking lozenges (or lozenges) containing a biotin antagonist. Placing
n such a lozenge in the goat, one can obtain a prolonged delivery of one or more antibiotic substances during a prolonged period of time. The use of these tablets is particularly recommended during rest and the example which will be described in detail below relates to a tablet lasting 8 hours, this period being widely regarded as a normal duration of one night's sleep. .

The basic data on which the calculations are based are as follows
saliva production without stimulation: 110 ml / h max;
biotin content of saliva: 0.8 microgranes / liter;
ratio of the effective biotin to biotin antagonist: 70 / I;
desired period of effectiveness: 8 hours.

 There are mainly two kinds of delay pads.

The first comprises a practically insoluble matrix from which the active principle is leached for a certain period of time. In the other, the whole tablet dissolves but slowly, the excipient and the active substance being released at the same time. The Applicant prefers the second category for its invention. US Patents Nos. 3,039,933 and 3,362,8SI describe the manufacture of this kind of pellet.

 A property common to both kinds of lozenges is that although ensuring a prolonged release of the active substance, it does not ensure constant release rates. In the case of tablets with a stable matrix, the reservoir of the active substance is gradually emptied. In the case of fully soluble pellets, the dimensions of the pellets decrease over time and since the surface area decreases, the same applies to the rate of dissolution. In both cases, this results in a gradual decrease in the rate of release of the active substance. In certain cases, this behavior is advantageous as with the drugs with which it is desired to establish a certain level of the active substance, then, simply to maintain levels which are much lower than the first. In the present invention, this situation is not sought and required countermeasures.

 To highlight the action that occurs during the dissolution of lozenges such as these, it is necessary to consider a delay lozenge of 8 hours which must oppose the action of biotin brought into the mouth with saliva .

At the rate of 110 ml / h and 0.8 micrograms of biotin / liter, the maximum 880 ml released will introduce 0.704 micrograms of biotin. With the ratio in'70 previously mentioned as a value of the biotin antagonist, it appears that it was advisable to incorporate 49.3 micrograms intended to be released during the period of â hours. However, in order for this quantity to be released at a constantly decreasing release rate, the average rate would only really exist in the middle of the time period with a double rate during the first half period and a half rate in the second.

 At first glance, it may seem that to remedy this situation, it would suffice to double the dosage of the tablet, but this is only a partial solution; indeed, the lozenge has dissolved all the same towards the end of the period of time and the rate of release is too low when the period approaches its end, unless there has been incorporated therein large excesses of the active substance .

This problem is solved by the present invention by using a lozenge for 16 hours which begins with a double dose of biotin antagonist. In this way, half the release rate of the tablet is reached just after 8 hours; before release rates are higher than actually needed, but protection is provided at all times. Thus, the lozenge would contain 98.6 micrograms of biotin antagonist and would be intended to be used for
16 hours, but would only really be used for 8. In fact, the best solution is to divide the dose between two lozenges and place one of them near each cheek at bedtime.

 It should be noted that such delay pastilles intended for use in the invention can also be produced for other durations, for example, to be used between meals. The dose is calculated for twice the amount necessary to oppose the action of the amount of biotin that can be released during the period of time that one wishes to cover, and the lozenge must dissolve in a period of time which is double that.

 It goes without saying that the invention> is in no way limited as regards the form of the pellets used to enclose the biotin antagonist. It is advantageous that the base of the tablet consists of a material such as carboxy-methyl-cellulose, polyacrylic acid @@ similar substances which do not easily constitute food for micro-organisms; natural gums often serve as food for bacteria and molds and are less advantageous. The Applicant also does not intend to limit the scope of its invention to the particular 16 hour delay tablet described above; indeed, a tablet with these multiples having a very high internal concentration of biotin antagonist and successively lower percentages in the external layers can ensure a more constant rate of liberation, but the increased cost of a more complex tablet should be considered when comparing it with the cost of using a slightly higher amount of biotin antimeta bolite to maintain efficacy, dance a pasti the simplest to make.

EXAMPLE 24
Delayed oral lozenge designed for an 8 hour effectiveness and containing an inactivating biotin antagonist.

Delay tablet base I00 grams
Avidin 4.93 mg
This composition is compressed to obtain pellets for I6 hours of I gram each.

 Two lozenges are placed in the mouth at bedtime, one near each cheek.

EXAMPLE 25
4 hour delay oral tablet intended to be used between meals and containing an antagonist of the biotin of the antimetabolite tire.

Delay tablet base 100 grams
Desthiobiotin 2.97 mg
This composition is compressed to obtain pellets for 8 hours and weighing I gram each.

 Two lozenges are placed in the mouth between meals, one near each cheek.

EXAMPLE 26
5 hour delay oral tablet containing biotin antagonists of both the inactivating and antimetabolite type.

100-gram retille base
Avidin 2.97 mg
Desthiobiotin 2.97 mg
This composition is compressed to obtain pellets with a duration of I6 hours weighing I gram each.

 Two lozenges are placed in the mouth at bedtime, one near each cheek.

 Considerations regarding possible health hazards that may result from the use of the invention.

 Since biotin is an essential vitamin for humans, consideration should be given to the possible effects on the supply and supply of this vitamin which could result from maximum use of the features of the present invention.

There are some significant factors that come into play that should be considered
I. The treatments only target the biotin present in the mouth. They are not directed to the body's biotin stores in order to decrease their levels in the mouth.

The quantities of biotin to be eliminated and the quantities of agents used for this purpose involve an ecosystem represented by less than 10 ml of saliva at a time, and the total production of a day is indicated as being at the maximum of 1500 ml both by the manual "The Biochemists'Handbook" (Van Nostrand I962) and by the book "Texbook of Biochemistry" by est & Todd (Mac Millan, I952). The first reference indicates as biotin content of saliva 0X8 microgram / liter. Even with a maximum of 2 liters of saliva the total amount of biotin rendered inoperative only amounts to 1.6 micrograms / day.

 Antagonists, even with overdoses of 100%, can therefore only cancel the action by a maximum of 3.2 micrograms of biotin / day.

2. The human body, which represents a much larger system, receives its biotin from two sources (Recommended Dietary Allowances, published by NAS I974)
at. food: I00 to 300 micrograms / day
b. normal intestinal microflora in sufficient proportions to present levels representing 3 to 6 times the amount from food to be eliminated as excess in the urine. Even by adopting the minimum biotin values of food, we get figures of 300 to 600 micrograms which are above the needs of the body.

By comparing the data (I) indicating the quantities of biotin which can be annihilated by the use of the invention: that is to say more than 3 micrograms, with (2) the biotin surpluses of 300 to 600 micrograms, it can reasonably be said that the risk of a possible lack of biotin is extremely unlikely.

 Other facts reinforce the impression of confidence in maintaining a satisfactory supply of biotin for the body. First, the NAS has not established any prescription for biotin because this vitamin is amply supplied by our diet. In an experiment by Sydenstricker (JAMA, in42, II8: II99-I200), it has been shown that subjects must be fed a diet comprising 30% egg white for seven weeks before observing the acute symptoms biotin deficiency; the dosage of avidin was massive enough to be able to inactivate around 900 micrograms of biotin / day.

 It is difficult to imagine that the possibility of depriving the body of less than 4 micrograms of biotin / day using the invention can produce similar effects.

Claims (26)

X E V E N D I C A T I O t; w I. Compositions for dental care chosen from the group comprising ires toothpaste; tooth powders, mouthwashes, chewing gum, confectionery, dental coating concentrates and delayed mouth tablets including optimal effective amounts of substances blocking the absorption of biotin, which have a negative effect on diet microorganisms whose diet requires biotin, microorganisms which are involved in the production of dental caries and in the formation of acid in the mouth, said substances being antagonists of biotin.  2. Compositions according to claim I, characterized in that they are in the form of a toothpaste.  3. Compositions according to claim I, characterized in that they are in the form of a toothpaste.  4. Compositions according to claim I, characterized in that they are in the form of a toothpaste.  5. Compositions according to claim I, characterized in that they are in the form of a mouthpiece.  6. Compositions according to claim I, characterized in that they are in the form of a chewing gum (chewing gum).  7. Compositions according to claim I, characterized in that they are in the form of confectionery.  8. Compositions according to claim I, characterized in that they are in the form of confectionery having an outer layer containing biotin.  9. Compositions according to claim I, characterized in that they are presented as a concentrate producing a dental coating.  IO. Compositions according to claim I, characterized in that they are in the form of oral delay tablets.  II. Compositions for dental care chosen from the group comprising toothpaste, dentifric powders, mouthwashes, chewing gum, confectionery, dental coating concentrates and oral tablets with prolonged or delayed dissolution comprising effective quantities of cptimal substances blocking the absorption of biotin, which have a negative effect on the diet of microorganisms whose diet requires bi tin, microorganisms which are involved in the production of dental caries and in the formation of acid in the mouth, said substances being inactivating biotin antagonists of the typ.  12. Compositions according to claim II, characterized in that they are in the form of a toothpaste  I3. Compositions according to claim II, characterized in that they are in the form of a fried paste.  I. Compositions according to claim II, characteristics in that they are in the form of a powder of tifrice.  15. Compositions according to claim II, characterized in that they are in the form of a mouthwash  I6. Compositions according to claim II, characterized in that they are in the form of a chewing gum.  I7. Compositions according to claim II, characterized in that they are in the form of confectionery.  I8. Compositions according to claim II, characterized in that they are in the form of confectionery having an outer layer containing biotin.  I9. Compositions according to claim II, characterized in that they press in like a concentrate producing a dental coating.  20. Compositions according to claim II, @ aractériises sees in that they are in the form of pastilles bu cales retrd.  CI Coirositijns for dental care selected from the group consisting of toothpaste, tooth powders, mouthwashes, chewing gems, confectionery, dental coating concentrates and oral tablets with prolonged or delayed dissolution including optimal effective amounts of substances blocking the absorption of biotin, which have a negative effect on the diet of microorganisms whose diet requires biotin, microorganisms which are involved in the production of dental caries and in the formation of acid in the mouth, said substances being inactivating type biotin antagonists.  22. Compositions according to claim 2I, characterized in that they come under the standard of a toothpaste  23. Compositions-according to revendi ation 21, characterized in that they are in the form of a toothpaste.  24. Compositions according to claim 21, characterized in that they are in the form of a toothpaste.  25. Compositions according to claim 21, characterized in that they are in the form of a mouthpiece.  26. Compositions according to claim 21, characterized in that they are in the form of a chewing gum.  27. Compositions according to claim 2I, characterized in that they are in the form of confectionery.  28. Compositions according to claim 21, characterized in that they are in the form of confectionery having an outer layer containing biotin.  29. Compositions according to claim 2I, characterized in that they are presented as a concentrate producing a dental coating.  30. Compositions according to claim 21, characterized in that they are in the form of oral delay lozenges.  31. Dental care compositions selected from the group consisting of toothpaste, tooth powders, mouthwashes, chews, cenriseries, dental coating concentrates and long-acting or delayed-dissolving oral lozenges including optimal effective amounts of substances blocking the absorption of biotin, which have a negative effect on the diet of microorganisms whose diet requires biotin microorganisms which are involved in the production of dental caries and in diacid formation in the mouth, said substances being biotin antagonists of both the inactivating type and the antimetabolite type.  32. Compositions according to claim 31, characterized in that they are in the form of a toothpaste  33. Compositions according to claim 31, characterized in that they are in the form of a toothpaste.  34. Compositions according to claim 31, characterized in that they are in the form of a toothpaste.  5. Compositions according to claim 31, characterized in that they are in the form of a mouthpiece.  36. Compositions according to claim 31, characterized in that they are in the form of a chewing gum.  37. Compositions according to claim 31, characterized in that they are in the form of confectionery.  38. Compositions according to claim 31, characterized in that they are in the form of confectionery having an outer layer containing biotin.  39. Compositions according to claim 31, characterized in that they are presented as a concentrate producing a dental coating.  40; Compositions according to claim 31, characterized in that they are in the form of delayed-release oral lozenges.