CZ302051B6 - Immunomodulating biological composition exhibiting regenerative and vitalization activity and process for preparing thereof - Google Patents

Abstract

In the present invention, there is disclosed an immunomodulating biological composition possessing regenerative and vitalization activity wherein the composition can be prepared using the following steps: (i) disintegration and homogenization of a mixture consisting of (a) lymph, cellular and body liquids and a liquid of cell cores of animals of the Lumbricus terrestris strain, (b) honey, (c) starch, (d) at least one monohydric, dihydric and/or trihydric alcohol containing in total 1 to 22 carbon atoms, (e) at least one aliphatic organic acid having in total 1 to 4 carbon atoms, and optionally (f) mumijo of vegetable-mineral origin whereby the ratio of the total amount of the components (b) and (c) to the total weight of the component (a) in the mixture ranges from 1 : 99 to 99 : 1; (ii) leaving the disintegrated and homogenized mixture at a temperature of 35 degC at the most and a pH ranging from 1.5 to 6.9 for a period of 1 to 24 hours; (iii) subsequent separation of supernatant from the mixture; and iv) fractionation of the supernatant and (v) its optional thickening and/or lyophilization.

Description

Technical field

The invention relates to an immunomodulatory biocomposition with a regenerative and vitalizing effect and to a process for its preparation.

BACKGROUND OF THE INVENTION

It is currently known to use various galenic forms based on the use of adaptogens. The most diverse plant and animal adaptogens have been known and used by humanity since time immemorial, but in recent years the knowledge and use of adaptogenic drugs have been unprecedented, new and new sources of active substances (sometimes discovered and exploited by man, sometimes unknown or neglected); there is an increasing hunger for the most diverse means of stimulating the body's defensive and healing powers and increasing its performance and adaptability to the burden of the external environment, as well as resistance to changes in the body's internal balance. Adaptogens - also stimulators (biostimulators), harmonizers, tonic - stimulate immunity (especially non-specific defense mechanisms of the organism) and metabolism are means that increase organism resistance, ability to withstand various types of load, adapt to faster or better changed conditions, ie better withstand stresses arising from increased demands on mental or physical performance, adverse climatic conditions, and overcoming difficulties that have been termed "civilization factors", etc., are characterized by a great versatility of preventive effect, especially in relatively healthy but exhausted individuals, or for persons exposed to the environment. Here, adaptogens put the whole organism into an efficient state without introducing anything new or foreign into the body and without exhausting it. They only preserve or restore the overall resistance and performance of the organism that is genetically intrinsic to it and mobilize its reserves. In doing so, they promote biosynthesis, i.e. the generation of the most important nucleic acids and protein compounds, and at the same time transfer of energy and their sources into the cell, thus creating the basis for their multilateral action. Adaptogens improve the overall condition of the organism, improve the condition of the nervous system, cells, tissues, accelerate and strengthen metabolism, assimilation and metabolism in general, accelerate the healing and regeneration processes, slow degenerative processes in the animal organism and thus aging processes tissues and cells, always have a positive effect on several specific functions at the same time, compensate for pathological fluctuations in the values of these functions and the content of certain substances in the body or help to improve these functions above standard, reduce inflammation; as a biogenic stimulator, it stimulates not only the physical but also, on several levels, the mental area; they often also have a booster effect - like roborans. Many adaptogens increase decreased muscle tone, but also partially remove excess tension and relieve spasm. They induce a sense of strength and good physical health, which is mostly based on real positive changes in the body caused by the adaptogenic drug. Because of their overall effect on health and subjective feeling and their effect in terms of stimulation of bodily, organ and cellular functions, they can be considered subtle, in some cases relatively aphrodisiacs. The term “harmonizers” used by some authors describes their ability to modulate mental and physical activity, better concentration, mental well-being, good CNS and all mental and physical functions, and improved performance and resilience. An important finding of some important adaptogens is that their efficacy does not decrease with long-term administration. Their distinctive feature is that they are generally not addictive. Adaptogens do not have to cause ionizing or stimulating effects on healthy people, which mainly occur in debilitated or diseased individuals. Adaptogens can provoke the production of interferon, which is a significant factor of immunity and inter alia slows down tumor growth. As adaptogens, resp. Drugs and preparations with adaptogenic effect are denoted by some types of plants and fungi (possibly also lower organisms) and raw, unprocessed vegetable, animal drugs in the form of raw materials or (exceptionally) also by mineral origin.

Drugs simply prepared (dried, cut, etc.) or directly active substances or complexes of these substances with adaptogenic action isolated from these drugs; however, preparations prepared from or with lower organisms and, in rare cases, raw materials and drugs of inorganic origin (eg some minerals rich in minerals) may also have an adaptogenic effect. Some raw or modified mixtures of substances of natural origin also act as adaptogens: royal jelly, propolis, bee pollen, snake venom (in our viper), mumio, complexes of substances in extracts or other preparations of organisms or their products (lower organisms, plants, fungi , animals) - e.g., Almykon (of mushrooms), Pangamin (sterilized spray yeast); JUVENIL in Sanoregen, Imuregen, JUWIM in Preventan, Fortisim (from bovine; they contain free amino acids, active ferrous compounds, oligopeptides and other substances mainly active in the immune system). Some animal hormones (growth, sex, etc.) also have a significant adaptogenic effect. Individual hormones have a relatively broad effect, on the other hand the individual components of the hormonal system are closely linked to each other and the hormonal system is in turn linked to the immune system and other adaptation mechanisms, so that the effect of the isolated hormone can be relatively broad. Many adaptogenic drugs have a proven antitumor effect, mostly in prevention, but often also in their own disease, while helping to endure the side effects of chemotherapy, surgery and radiation. Adaptogenes with antitumor effect are, for example: eleuterococcus and lentinan which is a polysaccharide. More detailed studies have shown that lentinan not only has an antitumor effect, but that it enhances immune processes and stimulates the histamine and serotonin sensitivity of the organism and thus has some adaptogenic effect. In addition to lentinan, glucans and muramyl dipeptide derivatives stimulate immunity, but thymus extracts, and interferons and some synthetic substances - levamisole as an anti-roundworm agent - have also been tested. Many of these substances have been used successfully. Chemists have isolated active substances from bacteria that activate immune cells and enhance certain steps of immune responses. The best-known is LPS (lipopolysaccharide), a complex molecule from the walls of intestinal bacteria, which is, however, highly toxic. However, most adaptogens have a preventive and curative anti-cancer effect (as a complementary treatment or as part of a comprehensive treatment). The disadvantage of small adaptogens is that they act as adaptogens in fewer indications, their effect is not so broad, and we have to take into account some restrictive conditions (eg because of their relatively higher toxicity); sometimes it is possible to perceive their effect as adaptogenic only within a relatively specific indication - eg in some cases in influencing blood sugar levels (although the drug in question is complex, the effect is specific). Isolated adaptogens are, for example, eleuterosides, aralosides, guaranin, gomisin, schizandrin. The most effective high-molecular substances - functional - plant biocatalysts are sometimes referred to by the collective name ergons (= agents, agents). This group includes vitamins, enzymes and hormones. By their effectiveness, they are important components of the adaptogenic effect of individual drugs. No one doubts the adaptogenic effect of vitamins today. The effect of all vitamins is intensified and prolonged in combination with other components of the adaptogenic effect - especially with certain other vitamins, trace elements, amino acids, etc.

Adaptogenic drugs can usually be combined with the use of vitamins, trace elements, enzymes and hormones in tablets for therapy and prevention, thus enhancing their effect. The sources for their industrial processing are, for example: papaya, pineapple, pancreas of young farm animals (papain, bromelain, trypsin, chymotrypsin enzymes; from them the combined preparations Wobenzym and Phlogenzym are made). Egg yolks contain coenzyme Q-10, also known from several pharmaceutical preparations. This enzyme is sometimes referred to as a source of cellular energy). The hormones in the plant kingdom are physiologically important sesquiterpenic (sesquiterpenoid) substances with cyclic structure, the most important of which is the group of triterpenic (triterpenoid) substances - triterpenes, including steroid and non-steroid substances. Steroids differ from other triterpenic substances in that they are tetracyclic. The basic hydrocarbon of the steroid compound group is cyclopentane hydrophenanthrene. It consists of fully hydrogenated phenanthrene and a condensation-attached cyanopropene ring. Non-steroidal triterpenes include, for example, cardioglycosides and some saponins. However, the group of steroid triterpenes is much more important in terms of its effect, mainly because it is one of the most active substances in the animal kingdom - steroid hormones and substances that are precursors or metabolites of these hormones and other substances similar in their chemical structure and composition and which

Because of this similarity, they can interfere with hormonal processes, either by directly affecting the production of animal hormones in the body or by imitating the effect of these animal hormones. Amongst steroid substances; These include sterols (chemically sterins; these are steroids having the character of single-function secondary alcohols), bile acids, some hormones, cardiac glycosides, and some saponins. Also sterols can be classified according to their occurrence into animal (zoosterols), plant (phytosterols, phytosterines) and sterols contained in fungi and fungi (mycosterols), which can imitate the effects of hormones in the plant and animal kingdom. For some substances in the bodies of simpler plants and animals, their effect seems to be a developmental precursor to the role of hormones and other ergons in developmentally higher organisms. Sometimes (eg ecdysones), insects use substances from the plant kingdom as a substitute for their own hormones. Sometimes the effect of small adaptogens does not seem as broad or pronounced as the drugs or preparations with this label would have. It should be noted, however, that in some cases, some diseases indicate a serious impairment of the body's natural adaptability and immunity, its immunity, and the metabolic balance. about certain destructive processes in the body. Therefore, often significant or even minor lasting effects such as blood sugar, allergies, asthma, eczema, malignancy, cyst formation and many other diseases, it may be a sign that we managed to improve the complex condition of the body, parameters of several important functions and an adaptogenic effect. Adaptogens and preparations prepared from them (Preventan, Juwin, Juvelin, Solcoseryl, etc.) based on dialysis and hydrolysis of bovine or calf blood have been used for a long time. The common disadvantages of the abovementioned adaptogenic support agents with a separate content of said active substances are the following. Since, as mentioned above, current preparations and compositions of plant and animal origin, based on adaptogenic action, cannot guarantee sufficient biological stability, a well-defined mechanism of action, perfect biological resorption, a precise chemical composition and concentration of active substances, there is an increased possibility allergic reactions and the effect of these substances cannot be exactly specified. The aforementioned drawbacks result in a very slow onset of the biological effect of adaptogenic substances and a relatively frequent recurrence of symptoms of the condition being treated.

SUMMARY OF THE INVENTION

The subject of the invention is an immunomodulatory biocomposition with a regenerative and vitalizing effect, obtainable by (i) disintegrating and homogenizing a mixture of (a) lymph, cellular and body fluids and cell nucleus fluid of Lutnbricus terrestris, (b) honey, (c) starch, (d) ) at least one monohydric, dibasic and / or tribasic alcohol having a total carbon number of 1 to 22, (e) at least one aliphatic organic acid having a total carbon number of 1 to 4 and optionally (f) mummia of plant-mineral origin, the weight of components (b) and (c) to the total weight of component (a) in said mixture is 1: 99 to 99: 1, (ii) leaving the disintegrated and homogenized mixture at a temperature of not more than 35 ° C and a pH of 1.5 to 6; 9 for 1 to 24 hours, (iii) subsequently separating the supernatant from the mixture, and (iv) fractionating the supernatant and (v) optionally concentrating and / or lyophilizing it.

Preferably, the above-described immunomodulatory biocomposition having a regenerative and vitalizing effect is obtainable by said method, wherein the mixture as component b) comprises natural honey.

Preferably, the above-described munomodulatory biocomposition with a regenerative and vitalizing effect is obtainable by said method, wherein the composition as component c) comprises natural starch.

Preferably, the above-described immunomodulatory biocomposition with a regenerative and vitalizing effect is obtainable by said method, wherein the composition as component f) comprises an mumio of the type Archartas.

The present invention also provides a process for the preparation of an immunomodulatory biocomposition as defined above having a regenerative and vitalizing effect, comprising preparing a mixture of (a) lymph, cell and body fluids and cell nucleus fluid of the genus Lumbricus ter-3EN 302051 B6 restris, (b) honey, (c) starch, (d) at least one monohydric, dibasic and / or tribasic alcohol having a total carbon number of 1 to 22, (e) at least one aliphatic organic acid having a total carbon number of 1 to 4 and optionally mumia of plant-mineral origin, wherein the ratio of the total weight of components (b) and (c) to the total weight of component (a) in the said mixture is 1: 99 to 99: 1, whereupon the mixture disintegrates and homogenizes and 35 ° C and pH 1.5 to 6.9 for 1 to 24 hours and then the supernatant is separated from the mixture, which is fractionated and optionally concentrated and / or lyophilizes.

Preferably, the supernatant is separated from the mixture by centrifugation. Preferably, the centrifuged supernatant is precipitated with acetate buffer, treated with magnesium and / or zinc salts, centrifuged, dissolved and filtered through a microfilter having a pore size of 2 to 60 microns. Preferably, the separated supernatant is concentrated to a dry matter content not more than 25 g / 100 ml of supernatant. Advantageously, excipients, in particular pharmaceutically acceptable excipients, are added to the supernatant or its lyophilisate.

In the preparation of the composition according to the invention, enzymolysis occurs, wherein the product of enzymolysis of lymph, cell and body and cell nucleus fluids of the Vermes animal of the family Lumbricidae contains as active ingredients essential amino acid complexons, free amino acids, oligopeptide complexons, phospholipids, polypeptides, bound Fe ++, low molecular peptides and polypeptides, enzymes and glycoproteins, enzymatic grafts, hyaluronic acid and its derivatives, collagen and lysozyme. As a source of essential amino acid complexes, free amino acids, oligopeptide complexons, phospholipid, polypeptides, suitably organically bound Fe ++, low molecular weight peptide and polypeptides, enzymes and glycoproteins, preferably this composition comprises complexons of essential amino acids, free amino acids, oligopeptide complexons, phospholipid, polypeptides, suitably organically bound Fe ++, low molecular weight lama peptides and polypeptides, enzymes and glycoproteins from animal bodies

Vermes, family Lumbricidae. As lymph, cell and body fluid and cell nucleus fluid of Vermes animals, the Lumbricidae family preferably this composition comprises lymph, cell and body fluid, and cell nucleus fluid from animals of the genus Lumbricus terrestris, natural honey and starch.

Overview of the drawings

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 shows a chromatogram obtained by the MALDI-TOF method and predetermining the determination of peptides in the range of relative molecular weights of 0 to 5000 Da in the biocomposition of the invention.

DETAILED DESCRIPTION OF THE INVENTION

Example 1

Preparation of the immunomodulatory biocomposition according to the invention in the form of a concentrated solution of kg of animals of the Vermes strain of the family Lumbricidae of the species Lumbricus terrestris is washed for 40 minutes in a solution of 20 liters of distilled water containing 240 g of sodium chloride. Then rinse with 10 liters of distilled water at a maximum temperature of 4 ° C. 1 g of maltose and 6.5 g of sodium caprylate, 15 g of sucrose are then added and disintegrated for 30 minutes. 135 liters of distilled water at 15 ° C are slowly added and disintegrated for a further 20 minutes. 55 liters of 96% ethyl alcohol are added, with stirring, and the pH is adjusted to 5.38 with acetic acid. 120 grams of concentrated mummy of plant-mineral origin Acharbags, 17 500 grams of natural honey and 650 grams of starch are added. Under constant homogenization at 25 ° C, enzymolysis was allowed to proceed for 12 hours. The solid slurry was centrifuged in a high speed centrifuge. The centrifuged supernatant was cooled to 8 ° C and precipitated with an acetate buffer

Stirring was continued at pH 5.5. Leave the precipitate to stir for 4 hours while lowering the temperature to -12 ° C. Centrifuge the precipitate in a high speed centrifuge. Dissolve the precipitate in 20 liters of distilled water at +4 ° C, adjust the pH to 4.2, add a technological amount of zinc oxide and magnesium carbonate and centrifuge the solution. To remove any residual solids with a very small particle size, the homogeneous solution obtained can be filtered on a pressure filter through a 4.5 micrometer filter insert. The supernatant can then be concentrated by vacuum.

io Example 2

Determination of the effect of the biocomposition according to the invention on certain skin diseases

The effect of the biocomposition according to the invention has been demonstrated in the treatment of certain skin conditions. The following results are shown in Table 1.

Table 1

Diagnosis Number of cases Evaluation Psorlase chronic 12 7x good 5x medium Palm Psoriasis Barber 6 2x good 4x medium Palm thyroid eczema 3 lx good 2x medium Neurodermitis 2 2x good Uchen planus 3 2x good lx weak Lower leg ischtyosis 2 2x medium Hyperpigmentation of the forehead 2 2x good Eczema atopkký 13 9x good 3x medium 1x without effect Eczema contact, allergy. 6 2x good 2x medium 1X without effect 1X < seborrheic dermatitis 4 2x good 2 x medium Uchen planus 2 2 x medium Prurigo nodul. Hyde 2 lx good lx medium Pruritus 2 2x good Acne vulgar 16 Ux good 3x weak 2x without effect Rosacea 6 5x good lx medium Dermatitis perioral 3 3x medium Dermatitis seborrheic 3 2x good lx medium Cold haze 1 lx very good Burn 1 lx very good

-5GB 302051 B6

Example 3

Determination of the effect of the biocomposition according to the invention on hemorrhoid diseases

As part of the demand, immunomodulatory biocomposition with regenerative and welcoming effect was tested as an active substance in the treatment of hemorrhoids. Two product forms were chosen, namely the form - suppository containing 5% immunomodulatory biocomposition with regenerative and welcoming effect and the form - cream with applicator containing 5% immunomodulatory biocomposition with regenerative and vitalizing effect.

The survey was conducted by the Canadian PROCTO-Clinic Sanatorium for its use in 1,440 patients. The results of this assay are shown in Table 2 below.

Table 2

Number patients State Species Application Length application Completely healed Retreat difficulties Beze changes 180 Acute bleeding external suppository 2x 1 169 10 1 180 Chronic bloodless external suppository 2x 1 178 2 - 180 Acute bleeding internal suppository 2x1 121 53 6 180 Chronic bloodless internal suppository 2x1 163 13 4 180 Acute bleeding external cream 2 x 1 in 20 ml 173 6 1 180 Chronic bloodless external cream 2 x 1 in 20 ml 177 2 1 180 Acute bleeding internal cream 2 x 1 in 20 ml 103 56 21 180 Chronic bloodless internal cream 2 x 1 in 20 ml 98 51 31

Total Completely healed Retreat difficulties Beze changes 1440 1182 193 65

Example 4

Determination of the effect of biocomposition according to the invention on wrinkles

Immunomodulatory biocomposition with regenerative and vitalizing effect has been tested on the basis of positive feedback from consumers to determine its effect on wrinkle smoothing and regeneration of affected skin. The product has been tested in several beauty salons and by users' statements.

-6GB 302051 B6

The results obtained are shown in Table 3 below.

Table 3

Number persons Skin type Skin condition Disappearance completely Disappearance of Part Ineffective Time necessarily application 116 normal neglected 98 15 Dec 3 1-21 days . 121 Dry maintained 109 11 1 1-15 days 100 ALIGN! oily maintained 97 3 - 1-15 days 113 normal maintained 111 2 - 1-15 days 106 dry neglected 94 6 6 1-21 days 125 dry / oily maintained 78 42 5 1-21 days 108 dry / oily maintained 101 6 1 1-21 days

Total number of persons Disappearance completely Disappearance of part Without changes 789 688 85 16

Example 5 io

Determination of the effect of the biocomposition according to the invention on striatum, cellulite, scars, swelling and hematomas.

Based on observed effects of immunomodulatory biocomposition with regenerative and vitalizing effect, it was tested in cooperation with several beauty salons, gynecological clinics, burn clinic, surgical clinic - Motol and rehabilitation institute Kladruby the effectiveness of immunomodulatory biocomposition with regenerative and vitalizing effect in form of cosmetic carrier products. , for skin damage in the form of stretch marks, cellulite, scars, swelling and hematomas.

The results obtained are shown in Table 4 below.

Table 4

Number Damage Age damage Completely vyhojeno Partly vyhojeno Without effect Time application 216 strije 1 week 215 1 - 14 days 209 strije 1 week 197 11 1 14 days 134 strije 1 year 98 30 6 6 months 129 strije 1 year 95 29 5 6 months 103 cellulite 1 week 97 4 2 2 months 106 cellulite 1 week 98 6 2 2 months 104 cellulite 1 year 77 20 May 7 6 months 105 cellulite l year 78 18 9 6 months 98 scars 1 week 93 4 1 6 months 94 scars 1 week 88 4 2 6 towns 100 ALIGN! scars 1 year or more 66 31 3 1 year 96 scars 1 year or more 61 29 6 1 year

-7EN 302051 B6

Number Damage Age damage Completely vyhojeno Partly vyhojeno Without effect Time application 55 edema 1 day 54 1 - 1 day 58 edema 1 day 51 6 1 1 day 61 edema 3 days 59 1 1 2 days 60 edema 3 days 53 5 2 2 days 78 hematomas 1 day 77 1 - 1 day 75 hematomas 1 day 70 4 1 l den 83 hematomas 3 days 81 1 1 2 days Sl hematomas 3 days 77 2 2 2 days

Example 6

Determination of effect of biocomposition according to the invention on burns, protection against intensive ultraviolet radiation and removal of pigment spots

The efficacy of the immunomodulatory biocomposition with regenerative and vitalizing effects contained in the preparations with cosmetic carrier was tested in cooperation with the participants of climbing and alpine expeditions in the Himalayas and Andes, the burn clinic, radiation treatment department, Kladruby rehabilitation clinic, solarium and beauty salons. Effects on burn healing, skin protection against high UV exposure and removal of unwanted pigmentation were tested.

The results obtained are shown in Table 5 below.

Table 5

Number Damage Age damage Completely vyhojeno Partly vyhojeno Without effect Length application 114 burns freshly 111 2 1 3 days 108 burns freshly 107 1 - 3 days 62 burns older than 24 hours (57) 4 1 Five days 69 burns older than 24 hours 63 4 2 Five days 33 UV Freshly 11 21 1 3 weeks 33 UV freshly 32 1 - 3 weeks 96 pigmentové spots - 94 1 1 8 weeks 101 pigmentové spots - 98 2 1 8 weeks

-8EN 302051 B6

Example 7

Determination of the effect of the biocomposition according to the invention on the hair cover

In experiments with immunomodulatory biocomposition with regenerative and vitalizing effect, it has been found, based on research, that the composite complex, in addition to congestion, strengthens and stimulates cell tissue and accelerates its regeneration and reproduction. Therefore, the immunomodulatory biocomposition with regenerative and vitalizing effect was applied in the form of shampoo in cooperation with oncology clinic, hairdressers and hair studies. According to the observations, the hair is prepared by amino acids and protein grafts, which causes the hair to become stronger and firmer, while at the same time leading to increased nutrition of the hair follicles, resulting in accelerated growth and improved hair condition. We consider the numerous reports of consumers that new hair grows after the product is subjective and very difficult to verify.

The results obtained are shown in Table 6 below.

Table 6

Number Preparation Very strong effect Partial effect Without effect Length application 250 shampoo + biocomposition 248 2 - 1 washing 226 Shampoo + bio-composition 225 1 1 intrusion into washed hair

Example 8

Determination of the effect of biocomposition according to the invention on periodontitis

In order to verify the possible effects of an immunomodulatory biocomposition with a regenerative and vitalizing effect and for its above-mentioned properties, we asked several dentists to test the effect of the aforementioned composite on periodontal tissues and similar inflammatory oral diseases.

The results obtained are shown in Table 7 below.

-9EN 302051 B6

Table 7

Number Damage Tent damage Completely vyhojeno Partly vyhojeno Without effect Length application 56 paradentosa morning 50 4 2 1 month 48 paradentosa morning 41 5 2 1 month 53 paradentosa late 43 5 5 1 month 55 paradentosa late 39 9 7 1 month 104 dental caries morning - 31 73 1 month 108 dental caries late - 13 95 1 month 44 aphtha morning 41 2 1 2 days 43 aphtha morning 39 3 1 2 days

Example 9

Determination of the effect of the biocomposition according to the invention in the eradication of HPV infection and poorly healing and skin defects

Immunomodulatory biocomposition with regenerative and vitalizing effect was tested in gynecological and obstetric center Brno, MUDr. Šárka Kutálková for HPV viral infection in the form of a sitting bath. 25% aqueous solution was tested at the Motol Hospital Surgical Department for infected non-healing pressure ulcers, postoperative wounds and leg ulcers.

The results obtained are shown in Table 8 below.

Table 8

Number Damage Age damage Completely vyhojeno Partly vyhojeno Without effect Length application 15 Dec HPV morning 14 1 - 1 month 12 HPV late 7 3 2 1 month 16 postpartum defects morning 14 1 1 7 days ^{15 Dec} postpartum defects late 9 3 2 7 days 22nd decubitus morning 19 Dec 1 1 14 days 21 decubitus late 18 2 1 14 days 30 post-operative early morning 29 1 - Five days 35 leg ulcers morning 29 5 1 14 days 32 leg ulcers late 26 4 2 14 days 8 leg ulcers with necrotic disintegration late 4 1 3 14 days

- 10GB 302051 B6

Example 10

Determination of the effect of biocomposition according to the invention on diseases of the digestive tract

As part of trials of the extension of possible applications of immunomodulatory biocomposition with regenerative and vitalizing effect, in cooperation with several private internal doctors, we tested the effectiveness of immunomodulatory biocomposition with regenerative and vitalizing effect on diseases of the gastrointestinal tract, namely duodenal ulcers and intestinal polyps.

The results obtained are shown in Table 9 below.

Table 9

Number Damage Age damage Completely vyhojeno Partly vyhojeno Without effect Length application 11 duodenal ulcer acute 8 2 1 3 weeks 12 duodenal ulcer chronic 5 5 2 3 weeks 9 polyps acute bleeding 7 2 - 14 days 10 poiypy chronic 6 2 2 14 days

Example 11

Comparison of tests and synergism of immunomodulatory biocomposition and concentrate of bioactive composition Vermio according to Czech patent No. 288509, according to prescribed methodology of SÚKL, SOP - NRL / M-1401:

Table 10

time 30 min 60 min 120 min concentration 0 -1 -2 0 -1 -2 0 -1 -2 Immunomodulatory biocomposition 1 according to invention 1:50 -t-H- ++ + + - - - - Immunomodulatory biocomposition 2 according to invention 1: 25 +++ ++ + + - - - - 11 -1 -2 -3 -4 -5 -6 -7 -8 -9 1. control + + - - - - - - - 2. control ++ + - - - - - - -

- II CZ 302051 B6

time 30 min 60 min 120 min concentration 0 -I -2 0 -1 -2 0 -1 -2 Bioactive ++++++ 74 ++ ++++ 88 ++++ ++ 58 composite Vermio Al according to U.S. Pat. CZ288509 1. 50 Bioactive ++++++ iui 44 ++++++ ++++ ++ (+++) (+++) ++ 42 composite Vermio B2 according to U.S. Pat. CZ288509 1:25 -1 -2 -3 -4-5-6 -7 -8 -9 1. control + -H- ++ + + + - 2.kontrola + -H- ++ + + + -

Explanation of symbols:

Values 0 to -9 determine the reading times (30 to -1, etc.) in a given time period.

+ number of crosses indicate the number of bacteria tested when reading the number of bacteria on the normoplase.

The cause of this synergism is not yet well understood, and although there is no attempt to link the essence of the solution to a particular theory of the mechanism of said synergistic effect, it is believed that the seven major active agents interact with each other to form more easily resorbable and more stable complexes. Although the most pronounced synergy can be observed when both active substances are present in the same order of magnitude, there is a relatively strong synergy even when one active substance is present in units of volume percent and the other is present in tens of volumes. percent. From this fact, it is clear that the two active substances exert a catalytically mediating effect on each other, which means that the two active substances catalyze each other during the process and subsequently when resorbed in situ in the organism of the animal being treated.

Example 12

Analysis of the composition of the biocomposition according to the invention

Sample: Immunomodulatory composition (dark brown liquid)

Subject of analysis: total protein of free amino acid total amino acid

- 12GB 302051 B6 iron oligopeptides phospholipids

Result of analyzes:

Sample dry matter: 14.97% wt.

The dry weight was determined after drying the sample at 105 ° C to constant weight.

Determination of iron

The sample contained 63 mg Fe / kg sample.

Iron was determined after mineralization of the sample with nitric acid and perchloric acid by the AAS method. Due to the nature of the sample, it was not possible to determine the valence of Fe.

Determination of nitrogen and crude proteins

The sample contained 8400 mg N / kg of sample, corresponding to 52500 mg of crude protein / kg of sample.

The nitrogen determination was carried out according to the Kjehldal method and was converted to a crude protein content by multiplying by a factor of 6.25.

Determination of polypeptides

Peptides were determined by the MALDI-TOF method, which allows determination of the relative molecular weight of individual peptides.

Peptides were dominant in the range of 500 Da to 17 kDa:

619.1 Da; 1385.1 Da; 2748.0 Da; 3163.7 Da ;, 3216.0 Da; 5090,6 Da

In addition to the above peptides, the sample contained a relatively rich spectrum of other peptides, but at lower concentrations (see the chromatogram in Figure 1).

Determination of phospholipids

Phospholipids were determined after extraction of the Hpidic fraction from the sample as phosphorus and converted to stearoyl-oleylglycerophosphatidylcholine.

The sample contained phosphorus in an amount of 2.14 mg / kg, corresponding to a phospholipid concentration of 54.6 mg / kg of the original sample. The lipid fraction was 6200 mg / kg of the sample, the proportion of phospholipids in the lipid fraction was relatively low, only 0.88% by weight.

-13GB 302051 B6

Determination of amino acids

AK AK concentration [mg / 10Og] Free AK Total AK Asp 29.70 167 Ser 16.50 302 GIu 88.80 235 Gly 11.10 278 His 32.30 433 Arg 64.30 432 Thr 14.00 689 Ala 30.70 301 For 58.40 279 Cys 16.50 18 Tyr 361.2 622 Wall 37.50 78 Met 6,500 15 Dec Ly. 36.80 44 He 15.60 105 Lea 17.90 630 Phe 13.40 1253 ΣΑΚ 851 2 5880

Determination of free amino acids was performed after previous deproteinization of the sample.

Total amino acids were determined after previous hydrolysis of the sample with hydrochloric acid (c (HCl) = 6 mol / l).

The content of individual free and total amino acids was expressed in mg AK / 100 g of the original sample.

Example 13

Microbiological determination of biocomposition according to the invention Strains used: Staphylococcus aureus, Streptococcus pyogenes

Place of collection:

Procedure: A 15% concentration of the preparation in distilled water was prepared in the pharmacy / a slurry was formed, and the preparation was dissolved in HCl. The HCl concentration was chosen to match the pH of the stomach. A homogeneous brownish suspension was formed and filtered.

- 14GB 302051 B6

Both the suspension and HCl alone were transferred to the microtiter plate at 0.25 μί, and a 0.25 μί 0.5 scale suspension according to the Mc Farland Staphylococcus aureus and Streptococcus pyogenes strains was added. After proper mixing and shaking, the microplate was incubated for 24 hours at 37 ° C.

After 24 hours, the mixtures were seeded on blood agar which was incubated for 24 hours at 37 ° C.

Growth was assessed after 24 hours.

Culture result: suspension with rain, water - negative / microbes did not grow, suspension with HCl - negative / microbes did not grow, HCl alone - negative / microbes did not grow,

Preparations were also prepared from the mixture, which were stained according to Gram and then examined microscopically.

Result of microscopy: suspension with distilled water - microbes not found suspensions with HCl - microbes not found HCl alone - microbes not found

In general, a novel immunomodulatory biocomposition with a regenerative and welcoming effect utilizes biochemical reactions to obtain new, more efficient and more stable biologically active substances as compared to those originally contained in separately active substances. This is further supported by the method for the preparation of said immunomodulatory biocomposition with a regenerative and vitalizing effect, according to the method of the invention. The inorganic and organic constituents contained in the purified mummy of plant-mineral origin accelerate the process of enzymolysis and cleavage of hemoglobin to form a heme and pyrrole nuclei, while facilitating the leaching and stability of enzymes from cells of the Lumbricidae family. The balsamics contained in the mummies form esters with unsaturated fatty acids, thereby accelerating their resorption and breaking down in the body during reaction with free radicals. Disintegrated and stabilized organisms of the Lumbricidae family, Vermes, will release large amounts of enzymes, including other active substances, during disintegration, and will accelerate, with a significant increase in efficacy, the enzymolysis of lymph, cell and body fluids and cell nucleus fluids of the Vermes to form the following components.

Essential amino acid complexes in a suitable composition are initiators of endocrine gland hyperfunction, especially the pituitary gland, which is catalytically provoked to increase neurohormone production, particularly a number of endorphins. These neurohormones then act on the CNS, both as a natural tonic, which explains the application of substantial sedation, the elimination of depressive states, insomnia, fatigue syndrome and the like. At the same time, however, pituitary products are regulators of the whole body's energy system, through the neurohumoral apparatus of the body, thus ensuring equalization of energy potentials in the individual governing bodies of the whole organism. This effect is compared to internal reflexotherapy and brings harmonizing effects on internal organ functions. This explains the effects of the above preparation on blood pressure balancing, stabilizing the blood count, increasing muscle tone and increasing recent memory.

Oligopeptide complexons initiate the thymus function, which is the producer of T lymphocytes and also the spleen, which supplies blood to B lymphocytes, both of which are one of the basic components of the body's defense system. This factor is one of the decisive factors in the relatively rapid increase in the body's immune response.

- 15 GB 302051 B6

Phospholipids in the immunogenic system are characterized in the membrane-toxic surfactant group. The membrane-toxicity of this phospholipid agglomerate consists in blocking the membrane / receptor / cell lipoprotein channels and thereby creating a barrier to nutrition and oxygen supply to the cell. It is an immune response such as a bacterial, viral, etc. attacking a damaged cell that is dying by the immune system.

The peptide-to-polypeptide component belongs to the group of substances acting as an impulse immunological response. The intake of such a modified and acceptable protein causes the production of antigens that function as a defense system against the pathology of the organism. In addition, significant levels of lysoxin and lysozyme have been shown in experimental blood applications, leading to the consideration that oligopeptides, along with other predominantly low-molecular-weight peptide products, as the cornerstones of the formation of these "non-specific antibiotics" are initiators of their self-formation , which creates an additional factor in the body to strengthen the body's defense against external noxas.

Suitably organically bound Fe ++ resulting from the decomposition of porphyrin chains into pyrrole nuclei and Fe ++ cation strengthens the blood pool between the scythe and the spleen, thereby improving hematopoiesis.

Low molecular weight peptides and polypeptides act on the regenerative ability of the cellular system, and this effect is manifested by the extremely accelerated healing of traumatic and postoperative conditions. According to the latest research, these substances showed very significant antibacterial and antiviral properties.

Enzymes and glycoproteins and their function in the body have been extensively described in a large number of scientific papers.

All these factors give a characteristic feature of polyvalence in its action, and thus are characterized by its non-specific effects according to the condition of each organism. This is the cause of the variety of effects in its use, where it is best portrayed as an antibacterial, antiviral, harmonizing and preventive, as it puts the organism in an optimal state and increases the defenses as such. The indisputable advantage of long-term application is both mentally and therapeutically preventive, its trouble-free tolerance, non-toxicity, immunomodulatory ability, antibacterial and antivirotic action and regulatory properties associated with improving the body condition.

In view of the above, the newly designed immunomodulatory biocomposite with a regenerative and vitalizing effect, in contrast to the separate components it contains, has a more pronounced healing, stabilizing, soothing, regenerative, antibacterial and antiviral effect, while a very strong immunomodulatory effect is still present. .

In the preparation of the newly proposed immunomodulatory biocomposition with a regenerative and vitalizing effect, a mixture of blood, fertilizer, milk, californian and earthworm (Lumbricus terrestris.) Is preferably used, in which the above synergy is most pronounced. For disintegration of Lumbricidae bodies and homogenization, high-speed mixers with homogenizers are typically used. The aim of disintegration and homogenization is to achieve lesion to cell nuclei, which allows perfect release of cellular enzymes. Any physiologically acceptable monohydric, dibasic or tribasic alcohol having a total carbon number of 1 to 22 is added; preferably ethyl alcohol is used because of its good tolerability by the animal organism. After adjusting the pH to 1.5 to 6.9 with acetic acid, mumio, natural honey and starch are added. The mumio used is preferably an Archa-bag type mumio, especially for its favorable content of about 28 elements, 10 silicate groups, 6 amino acids, balsamics and essential oils. After completion of the enzymolysis reaction to the mixture of desired products, centrifugation is performed on a high speed flow centrifuge from most solid particles. The supernatant obtained is precipitated with acetate buffer to a value of 1.9 to 5.8 at -15 to -1 ° C and centrifuged. Dissolve the precipitate in distilled water at +4 ° C, adjust the pH to 1.5 to 6.9, add

- 16GB 302051 B6 Centrifuge the technological amount of zinc oxide and magnesium carbonate. To remove any residual solids with a very small particle size, the homogeneous solution obtained can be filtered through a microfilter with a corresponding pore size. The resulting homogeneous solution is concentrated in vacuo and optionally preferably lyophilized. The resulting homogeneous solution, or a lyophilizate thereof containing a novel combination of active ingredients, can be administered as such or formulated in the form of an appropriate formulation by the addition of appropriate excipients.

Claims (9)

PATENT CLAIMS 1. Immunomodulatory biocomposition with regenerative and vitalizing effect, ready (i) 15 disintegrating and homogenizing a mixture of (a) lymph, cell and body fluids and cell nucleus fluids of animals of the genus Lumbricus terrestris, (b) honey, (c) starch, (d) at least one monohydric, dibasic and / or tribasic alcohol with a total (e) at least one aliphatic organic acid having a total number of carbon atoms of 1 to 4 and optionally (f) a mummy of plant-mineral origin, wherein the ratio of the total weight of components (b) And (c) the total weight of component (a) in said mixture is 1: 99 to 99: 1, (ii) leaving the disintegrated and homogenized mixture at a temperature of not more than 35 ° C and a pH of 1.5 to 6.9 for 1 up to 24 hours, (iii) subsequently separating the supernatant from the mixture, and (iv) fractionating the supernatant and (v) optionally concentrating and / or lyophilizing it. 2. An immunomodulatory biocomposition having a regenerative and vitalizing effect according to claim 1, characterized in that the mixture comprises, as component b), natural honey. 2, characterized in that the mixture as component c) comprises natural starch. Immunomodulatory biocomposition with a regenerative and vitalizing effect according to claim 1 or Immunomodulatory biocomposition with a regenerative and vitalizing effect according to any one of claims 1 to 3, characterized in that the mixture as component (f) comprises an mumio of the type Archartas. 35 A method for the preparation of an immunomodulatory biocomposition with a regenerative and vitalizing effect according to claim 1, characterized in that a mixture of (a) lymph, cell and body fluids and cell nucleus fluid from animals of the genus Lumbricus terrestris, (b) honey, (c) starch (d) at least one monobasic, dibasic and / or tribasic alcohol having a total number of carbon atoms of 1 to 22, (e) at least one aliphatic organic acid having a total number of carbon atoms of 1 to 4 and optionally mummies of plant-mineral origin; the total weight of components (b) and (c) to the total weight of component (a) in said mixture is 1: 99 to 99: 1, after which the mixture is disintegrated and homogenized and left at a temperature of not more than 35 ° C 6.9 for 1 to 24 hours and then the supernatant is separated from the mixture, which is fractionated and optionally concentrated and / or lyophilized. Method according to claim 5, characterized in that the supernatant is separated from the mixture by centrifugation. Method according to claim 6, characterized in that the centrifuged supernatant is precipitated with an acetate buffer, treated with magnesium and / or zinc salts, centrifuged, dissolved and filtered through a microfilter with a pore size of 2 to 60 microns. Method according to one of Claims 5 to 7, characterized in that the separated supernatant is concentrated to a dry matter content of not more than 25 g / 100 ml of supernatant. - 17 GB 302051 B6 Method according to one of Claims 5 to 8, characterized in that the supernatant or its lyophilizate adds excipients, in particular pharmaceutically acceptable excipients.