FR2674755A1 - Novel immunostimulatory medicinal compositions - Google Patents

Abstract

The immunostimulatory medicinal compositions of the invention prove to have properties which are greatly enhanced relative to those of the prior art by virtue of the fact that they contain at least one immunostimulant combined with at least one micronutrient, the immunostimulants comprising bacterial elements (such as actual bacteria, fractions of microbes, bacterial ribosomes or glycoproteins and membrane proteoglycans), biological polypeptides and/or synthetic immunostimulants, the micronutrients comprising, for their part, trace elements, vitamins and/or essential amino acids.

Description

New immunostimulatory drug compositions
The present invention relates to novel immunostimulatory drug compositions.

Most immunostimulatory drugs currently available do not have, for the most part, a significant activity in vivo, sometimes have side effects as is the case for levamisole or many interferons, or see their activity quickly decrease as some factors However, the main immunostimulators belong to the following non-limiting classes:
- bacteria and derivatives more or less purified, by
example, fractions of various germs (Imocur
Laboratoires Fournier), glycoproteins extracted from
klebsiella pneumoniae (Biostim from Cassenne Laboratories),
or bacterial ribosomes and membrane proteoglycans
(Ribomunyl from Inava Laboratories);
- biological polypeptides: interferons (IFN),
lymphokines, thymic factors, immunoglobulins (Ig),
- synthetic immunostimulants: Imuthiol (diethyl
sodium thiocarbamate), Inosiplex (or Isoprinosine
Delalande Laboratories).

To this non-exhaustive list of types and according to recent studies, it is possible to add micronutrients which are considered to have immunostimulatory activities; we can mention among others:
trace elements or trace elements, such as
zinc, copper, iron, manganese or selenium;
vitamins, for example E, A, B6, B9 or omega;
amino acids such as alanine (Ala),
glycine (Gly), threonine (Thr) or aspartic acid
(Asp).

 The various types of conventional immunostimulants do not show sufficiently effective and versatile activity, and micronutrients do not appear to provide improved results with respect to them.

 Nevertheless, the essential properties of the main components to which the present invention is based are recalled.

 It is generally accepted that the micronutrients mentioned below are considered immunostimulants although experience shows that they behave more like immunomodulators.

1 Essential trace elements.

1.1 Zinc
It plays an important role in cell metabolism (cell division, replication and transcription of deoxyribonucleic acid (DNA) by integrating into enzymes such as DNA polymerase, its immune action is carried out at three levels:
at the level of the lymphoid tissue: its deficiency causes the reversible atrophy of the thymus, the spleen and the lymph nodes; its administration stimulates the secretion of thymulin;
at the level of cellular immunity: its deficiency in the blood reduces the lymphocyte count, especially T, the proliferative response of lymphocytes to antigens and mitogens, the activity of natural killer cells (natural killers or NK), while increasing cutaneous hypersensitivity, and cytotoxicity of spleen cells;
at the level of humoral immunity: its deficiency reduces the production of lymphokines and immunoglobulins IgG and IgM, and the secretion of thymulin in the serum.

1,2 Manganese
Mn2 + manganese not only promotes a normal production of antibodies, but is even indispensable, as well as for the normal activity of the enzymes nucleases and polymerases including DNA polymerase; it influences immunocompetence by action on neutrophilic functions and macrophages; 1,3 Selenium
It stimulates T-dependent antibody responses and delayed hypersensitivity responses; it increases the titre of the antibodies; it has an anti-inflammatory and anti-radical action.

1,4 Copper Cu2T
It promotes the efficacy, synthesis and release of interferon, as well as the mobility of phagocytic leukocytes, and stimulates antiinfective activity.

1.5 Iron Fe2 +
It also promotes the efficacy of interferon and its effect on free radicals.

2 Essential Vitamins 2.1 Fat-soluble Vitamins 2.11 Vitamin A
P-carotene stimulates antibody responses, lymphocyte proliferation and antiinfective resistance, both in frequency and severity; it exerts an antiradical effect and its immunological role is considered as capital.

2.12 Vitamin E
Α-Tocopherol also stimulates antibody responses and delayed hypersensitivity responses, lymphocyte proliferation and antiinfective resistance; he plays a rattle in the trapping of free radicals. Selenium co-factor, these two micronutrients potentiate their effects when administered simultaneously.

2,2 Water-soluble Vitamins 2,21 Vitamin B6
Pyridoxine favors the production of thymulin, the normalization of lymphocyte responses, the stimulation of delayed hypersensitivity responses and antibody responses, especially post-vaccination, which is especially important since some immune stimulants such as Biostim exert an effect similar to that of vaccines. Vitamin B6 is, in addition, the cofactor of zinc, so essential to its action.

2.22 Vitamin B9 or Bc or M
Folic acid normalizes cutaneous reactions to antigens and lymphocyte responses to mitogens; Its deficiency or that in folate, in particular in the subjects affected by AIDS, causes neurological disorders.

3 Essential amino acids 3.1 Alanine
It increases lymphocyte reproduction and contributes to thymic growth.

3,2 Glycine
In the form of dimethylglycine (DMG), it promotes the antibody response, and in particular, it multiplies by four this response to pneumococcal vaccines; it also increases the cellular immune response by stimulating the metabolism of white blood cells.

3.3 Threonine
It stimulates the production of T antibodies and the increase of immunoglobulins IgG and IgM; 3.4 Aspartic acid
Aspartic acid, especially in the form of aspartate, would promote proliferation and differentiation of the thymus and bone marrow; it stimulates survival after total irradiation with regeneration of erythrocyte producing organs, as well as the metabolism of DNA.

 It should be noted that although some associations are known to use a so-called cofactor component (zinc / vitamin B6, zinc / manganese, selenium / vitamin E, for example), and as noted above, In vivo results are generally not as high as those that could be expected based on in vitro experiments, or on the animal.

 However, experience has shown that, by a judicious choice of micronutrient and immunostimulant components, even conventional immunostimulants, a surprising synergistic effect could be achieved, resulting in particular in a potentiation of the effects of the various constituents and a greatly increased efficiency compared to to previous associations.

 To better understand the technical characteristics and the advantages of the present invention, embodiments will be described, it being understood that these are not limiting as to their mode of implementation and the applications that can be used. make.

 The compositions below are given by soft capsule.

COMPONENTS Example 1 Example 2 Example 3 Example 4
Glycoprotein Anhydrous 1 mg 2 mg
Bacterial fraction 20 mg 20 mg
Copper Cu2 + 2 mg 4 mg
Fe2 + iron 2 mg 8 mg
Manganese 2 mg 10 mg
Selenium 25 pg 200 pg
Zinc 10 mg 50 mg
Vitamin A 3000 IU 30000 Ul
Vitamin B6 0.8 mg 2 mg
Vitamin B9 15 mg 75 mg
Vitamin E 10 mg 40 mg
Alanine 40 mg 60 mg
Magnesium aspartate 50 mg 50 mg 100 mg
Dimethylglycine 50 mg 90 mg
Threonine 50 mg 50 mg 350 mg
excipients
The glycoprotein used in the above examples is preferably that extracted from Klebsella pneumoniae.
The bacterial fraction is constituted, preferably in freeze-dried form from Diplococcus pneumoniae,
Haemophilus influenzae, Klebsiella ozaenae, Klebsiella pneumoniae, Neisseria catharralis, Staphylococcus aureus,
Streptococcus viridans, Streptococcus pyogenes.

The compositions below are given per scored tablet.

COMPONENTS Example 5 Example 6 Example 7 Example 8
Inosine + acédobène
+ Dimepranol 500 mg 500mg
Ribosomes of
Klebsiella pneumoniae + Diplococcus pneumoniae + Streptococcus pyogenes + Hemophilus influenzae 3 mg 3 mg
Copper Cu2 + 2 mg 4 mg
Iron Fe2 + 2mg 8mg
Manganese 1 mg 8 mg
Selenium 10 μg 300 μg
Zinc 10 mg 100 mg
Vitamin A 300 IU 30000 IU
Vitamin B6 1 mg 3 mg
Vitamin B9 15 mg 75 mg
Vitamin E 4 mg 40 mg
Alanine 20 mg 60 mg
Magnesium aspartate 50 mg 50 mg 90 mg
Dimethylglycine 50 mg 90 mg
Threonine 50 mg 50 mg 350 mg
excipients
In the eight examples, the manufacturing protocols and the excipients can be of any conventional and adequate type for obtaining products in pharmaceutically acceptable dosage forms. These products can generally be administered at the rate of two unit doses per day taken together or separately. morning and evening for a month or possibly longer, depending on the case and at the discretion of the prescriber, according to the return to normal of the biological constants measuring the immune power.

 The following experiments were carried out on the compositions of the above eight examples in the form of solutions, water, or physiological saline for in vivo tests, a solution of glycoproteins extracted from Klebsiella pneumoniae and purified (reference A). , and a solution of extracts of germs associated and killed, then purified (reference B).

 First experiment: Stimulation of phagocytic properties of polynuclear and macrophages in mice.

 The solutions are administered for one month by adapting the daily doses in proportion to the weight of the four hundred and forty selected mice (forty per solution to be tested).

On the polynuclear ones, on the one hand, and the macrophages, on the other hand, one could make the following observations:
Solution A has a significantly greater activity than serum both on the ingestion of latex particles by cells and on their chemotaxis;
Solution B, although more active than serum, is less effective than solution A on the ingestion of latex particles, but approximately as effective on chemotaxis;
For each of the solutions corresponding to Examples 1 to 8, the efficacy is clearly superior in all cases, both on chemistry and on the ingestion of latex particles, and both for macrophages and polymorphonuclear leukocytes. . These results are shown in the summary table below.

 Second experiment: Study of lymphocyte proliferation in humans and the resulting immunity to cell mediation.

 The batches of forty mice of each of the solutions A, B, and Examples 1 to 8 show a greater proliferation than that of the control group, however, the solution A is a little more active than the solution B, but remaining significantly lower than those solutions corresponding to Examples 1 to 8, which shows an immunity to cell mediation resulting from the application of the compositions according to the invention.

 Third experiment: Protection of the mouse against infection induced by the injection.

 Haemophilus influenzae is injected into mice and deaths are counted. Saline results in 100% mortality (forty mice), solution A 27.5% of deaths (11 mice), solution B 35% of deaths (14 mice) faster than A and the resulting solutions Examples 1 to 8 averaged 7.5% of deaths (3 mice) relatively late, demonstrating an increase in immune response capacity.

These results can be summarized as shown in the following table where 0 = null + = weak ++ = average +++ = strong

<tb><SEP> Stimul. <SEP> properties <SEP> Prolifer. <SEP> Protection
<tb><SEP> phagocytic <SEP> of <SEP> lymphocyte. <SEP> c / the infected.
<Tb>

<SEP> polynucl. <SEP> and <SEP> macroph. <SEP> at <SEP> Haemophilus
<tb><SEP> Man <SEP>% <SEP> of <SEP> death
<tb><SEP> ingestion <SEP> chemotax.
<Tb>

<SEP> part.latex
<tb> control <SEP> 0 <SEP> 0 <SEP> 0 <SEP> 100%
<tb> solution <SEP> A <SEP> ++ <SEP> ++ <SEP> ++ <SEP> 27.5%
<tb> solution <SEP> B <SEP> + <SEP> ++ <SEP> + <SEP> 35%
<tb> sol.ex.1 <SEP> +++ <SEP> +++ <SEP><SEP> +++ <SEP> 7.5%
<tb> sol.ex.2 <SEP> +++ <SEP><SEP> +++ <SEP><SEP> +++ <SEP> 7.5%
<tb> sol.ex.3 <SEP> +++ <SEP> +++ <SEP> +++ <SEP> 5%
<tb> sol.ex.4 <SEP> +++ <SEP> +++ <SEP> +++ <SEP> 10%
<tb> sol.ex.5 <SEP> +++ <SEP> +++ <SEP> +++ <SEP> 5%
<tb> sol.ex.6 <SEP> I <SEP> +++ <SEP> +++ <SEP> +++ <SEP> 7.5%
<tb> sol.ex.7 <SEP> +++ <SEP> +++ <SEP> +++ <SEP> 7.5%
<tb> sol.ex.8 <SEP> +++ <SEP> +++ <SEP> +++ <SEP> 10%
<Tb>
In fact experience shows that we obtain convincing results in the following ranges of contents:
Glycoprotein anhydrous 0.5 to 2 mg
Bacterial fraction of 10 to 50 mg
Inosine + acédobène
+ dimepranol from 100 to 900 mg
Ribosomes of
Klebsiella pneumoniae + Diplococcus pneumoniae + Streptococcus pyogenes + Hemophilus influenzae from 1 to 20 mg
Cu2 + copper from 1 to 10 mg
Fe2 + iron from 1 to 10 mg
Manganese 0.5 to 10 mg
Selenium from 10 to 300 pg
Zinc from 5 to 150 mg
Vitamin A from 300 to 40,000 IU
Vitamin B6 0.5 to 5 mg
Vitamin B9 5 to 100 mg
Vitamin E from 2 to 150 mg
Alanine 10 to 500 mg
Magnesium aspartate from 10 to 500 mg
Dimethylglycine 10 to 250 mg
Threonine 10 to 400 mg
The immunostimulatory drug compositions according to the invention thus have very strong properties compared to those of the prior art, because they contain at least one immunostimulator associated with at least one micronutrient, the immunostimulators comprising bacterial elements. (such as bacteria themselves, bacterial fractions, bacterial glycoproteins or ribosomes and membrane proteoglycans), biological polypeptides and / or synthetic immunostimulants, the micronutrients including, for their part, trace elements, vitamins and / or essential amino acids.

Claims (2)

An immunostimulatory drug composition with enhanced properties characterized in that at least one immunostimulator is associated with at least one micronutrient; 2 Composition according to claim 1 characterized in that the immunostimulators comprise bacterial elements; 3 Composition according to claim 2 characterized in that the bacterial elements are selected from the bacteria themselves, bacterial fractions, glycoproteins or bacterial ribosomes and membrane proteoglycans; 4 Composition according to one of claims 2 or 3 characterized in that the immunostimulators comprise biological polypeptides; Composition according to one of Claims 2 to 4, characterized in that the immunostimulators comprise synthetic immunostimulants; 6 Composition according to one of claims 1 to 5 characterized in that the micronutrients include trace elements; 7 Composition according to one of claims 1 to 6 characterized in that the micronutrients include vitamins; 8 Composition according to one of claims 1 to 7 characterized in that the micronutrients comprise essential amino acids;  9 Composition according to one of claims 1 to 8 characterized in that it contains at least two of the following constituents in the range of contents indicated per unit: Anhydrous glycoprotein 0.5 to 2 mg Bacterial fraction of 10 to 50 mg Inosine + acédobène  + dimepranol from 100 to 900 mg Ribosomes of  Klebsiella pneumoniae + Diplococcus pneumoniae + Streptococcus pyogenes + Hemophilus influenzae from 1 to 20 mg Cu2 + copper from 1 to 10 mg Fe2 + iron from 1 to 10 mg Manganese 0.5 to 10 mg Selenium from 10 to 300 pg Zinc from 5 to 150 mg Vitamin A from 300 to 40,000 IU Vitamin B6 0.5 to 5 mg Vitamin B9 5 to 100 mg Vitamin E from 2 to 150 mg Alanine 10 to 500 mg Magnesium aspartate from 10 to 500 mg Dimethylglycine 10 to 250 mg Threonine 10 to 400 mg