JP2847204B2 - Utilization of Fusarium fungal strains as producers and their adaptogen and base-based production with immunomodulatory effects - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to mycology and medicine, in particular to fungal-producing strains of physiologically active substances, and based on their bases, a production process for public health. About.

BACKGROUND OF THE INVENTION Inonotus obliquus [Inonotus]
Obliquis strains and bifungin products made on the basis of their bases are well known to those skilled in the art.

Bifungin is an extract from the swelling that grows on birch trees under the effect of this fungus. The product is a semi-flow containing 30% dry matter and 1% cobalt chloride or 1.5% cobalt sulfate added. It is used as a symptomatic agent to improve the health of patients with predominant flaccid gastrointestinal tract gastritis and dyskinesia (M.
D.Mashkovsky, Medical Agents, “Meditsina” Publisher
s, Moscow, 1984, Vol. 2, p. 162).

However, the resources of untreated birch trees for the production of bifungin are very low, comparable to the spectrophysiological effects of production.

Lentinas edodes, Ganoderma lucidum, and Pleurotis ostreatus are
Creates antiscleral effect, affects cholesterol content and normalizes levels in the body (Lee Khva Rei, Vasiliyev
LV, Orekhov LN, Tutya VA “Antisckerotic Proper
ties of Higher Fungi ", Problems of Alimentation No.
1,1989, pp16-20).

However, these cells are characterized by a low growth rate (mycelium growth is 5-7 days and in fruit bodies more than 70 days), and the physiological effect is a narrow area.

Fusarium cells cultivated on the basis of modern biotechnology grow at a high rate.

In the prior art, the fungus cells Fusarium sambiumium Fuckel varieties, Oshikaramu (Berk et al. Curt), Bulai V
The strain of SB-917 contains food producing bacteria and Hooder protein (A
ll-Union Collection of Industrial Microorganisms
of the VNIIgenetika Institute, collection code VKPM
F-169).

Previously, this strain was known as Polyporus sp.
917 (Microbiol, Science, 1986, v.3, No. 6, pp168-171)
It is called.

Bacterial Fusarium Sanbusium VSB-917, F-169
Has the following culture and physiological properties: On oat agar, it forms a white colony that turns pink on maturation and does not mature: commissures were not found.
It is characterized by two types of saponification: crescent-shaped conidia and Chamidospores with 2-5 distribution compartments. Cultured in a liquid nutrient medium forms multifilamentous mycelial mycelium with cyst distribution compartments and Voronin bodies. In the active growth stage, the mycelium is 2.5-3.5 microns thick, mycelial cells are homogeneous, free of lipids and high content, and the mycelium (mycelium)
Are rounded on average. Upon aging, the cells
It comes to contain many fats.

This strain is aerobic and in the temperature range of 22-28 ° C, 4.
It grows in the pH range of 5 to 7.0, with the optimal pH approaching 5.8.
Do not dilute gelatin. Glucose, saccharose, lactose, maltose, galactose, raffinose and starch, ethanol, glycerin, acetic acid, mannitol,
Digest xylitol.

For nitrogen sources, digest ammonium salts, nitrates, urea, and peptone.

Under the conditions of the submerged culture, the strain grows well on milk white (4-6% lactose) with 20-25 g ADM (absolutely dry) per liter of nutrient culture.

The output of the drain addition method is 1.4 to 1.6 kg / cu m / h.

The mycelium of the strain produced by the submerged culture method is 26-2% on a nutrient medium containing 4% by weight of ammonium nitrate, 0.3% by weight of potassium acid phosphate and 0.2% by weight of tap water.
Grow at a temperature of 8 ° C. for a culture time of 16 hours. The mycelium is autolysed and the resulting autolysate is 0.25-1.
0% in a nutrient medium, and tolulopsis Candida yeast (USSR No.
In 1034401, for 18.03.82 C12 No. 1/16), introduced
Its concentration in the culture medium should be 3 per liter of absolute dry matter.
From 1.1-32g to 34-44g.

The new application of the fungus Fusarium Sambusium VSB-917 is by using it as a producing bacterium of a physiologically active substance for preventive and therapeutic medicine.

DISCLOSURE OF THE INVENTION The main object of the present invention is to use the strain Fusarium Sambusium VSB-917 as a substrate for preparing a pharmaceutical product having a wide range of physiological effects on a living body and having no side effects even when administered. Is to do. According to the present invention, in order to achieve the above object, fungal strain Fusarium sambucinum Fuckel
Variant Bulai VSB-917 (the VNI Igentika Inst
Deposit code VKPM (F-169) for all union deposit collections of itute
Can be used to normalize cholesterol in patients suffering from cardiovascular disease from lipid metabolism disorder, induce triglyceride in serum, reduce the risk of progression of ischemic heart disease, Increases organ tolerance and serves as an adaptation source for protection against radiation exposure, is good for increasing leukocyte counts in leukemia, reduces abnormalities in the autoimmune process, mainly in the gastrointestinal tract, and A composition for the prevention or treatment of diseases due to metabolic disorders, indications and immunological effects, which can serve as a hyposensitivity agent for bacterial allergens. In addition, the present invention relates to 60-63% by weight of total protein (N × 6.25) carbohydrates 10-63% by weight lipids 4-8% by weight nucleic acids 3-6% by weight mineral substances 6-8% by weight vitamins 2.6-3.1% by weight water Cardiovascular disease caused by impaired lipid metabolism, produced by dehydrating a culture medium formed by submerging a single culture of the fungus strain Fusarium sambucinum Fuckel with the remaining composition in a liquid phase. Can normalize cholesterol, induce triglyceride in serum, reduce the risk of progression of ischemic heart disease, increase organ tolerance, and serve as an indication for protection against radiation exposure in affected patients In leukemia, it is good for the increase of white blood cell count, reduces the abnormal progression of the autoimmune process, mainly in the gastrointestinal tract, and as a hyposensitivity drug against bacterial allergens It can serve, metabolic disorders, provides a process for the preparation of a medicament for the prevention or treatment of diseases caused by adaptive Hara and immune effects owned. Further, the compositions can be manufactured in the form of tablets, granules, powders. In addition, the carrier of the product can be various foods.

Products based on the above-mentioned cells, over a wide range,
Has therapeutic and prophylactic effects. It contains a number of physiologically active substances, such as amino acids, immunomodulating polysaccharides, essential fatty acids, biological amines, ubiquinones, vitamins and naturally related microelements. Because you do. The product normalizes impaired metabolic processes in the body that are subject to cardiovascular disorders, obesity, diabetes, immune deficiency disorders, and vitamin deficiencies. The product eliminates allergic reactions, can be combined well with conventional therapeutics, enhances its effect on the body, and reduces unpleasant side effects, especially when radiation therapy is given to tumor patients. is there.

The chemical composition of the product is studied and the potential pathogenicity, acute and chronic toxicity,
Teratogenicity, carcinogenicity and co-carcinogenicity were studied. The study was conducted in line with the WHO requirements for testing microbial sources for irritation. The application of the product under the trade name "Milaif" was carried out on the basis of a series of experiments with the consent of the public health authorities.

Chemical composition of "Milaif"; wt% Total protein (N x 6.25) 60-63 wt% Authentic protein 42-46 wt% Carbohydrate 10-63 wt% Lipid 4-8 wt% Nucleic acid 3-6 wt% Mineral Substance 6-8% by weight Vitamin 2.6-3.1% by weight Water Rest The protein components of "Milife" are all essential amino acids (% of total protein): lysine, 9.2; isoleucine, 3.4; leucine, 4.5; phenylalanine + tyrosine , 4.5; cystine + methionine, 2.3; threonine, 3.
7; tryptophan, 0.8; valine, 3.6 and account for 35-45% of the total amino acids. For this metric,
"Milife" resembles a muscle protein that also contains 45% of essential amino acids. Protein substances and their derivatives
It is known to play an important role in the metabolic processes of digestion, respiration, excretion and exercise. Also, so-called "essential" amino acids must enter into it as a complete substance since they are not synthesized by the human body.

The carbohydrate portion of "Mylife", which amounts to 10-13% in dry form, is represented by 1-3-β-D-glycans, which exhibit immunomodulatory activity [MAKashkin, NGElin
ov (M.A. Kaskin, N.G. Elinoff):
"Immunomodulati activity of polysaccharides from fungi (Immunomodulati
ng Activity of Polysaccharides from Fungi), "Mycolo
gy and Phytopathology, Vol. 19, No. 4, pp. 1-3. The monomer composition of these glycans is 1.0: 1.38:
Represented by glucose, galactose, mannose and fructose in a 1.79: 0.5 relationship.

The lipid fraction of "Milife", which amounts to 4-8% in dry form, contains various forms of compounds, namely physiologically active substances consisting of phospholipids, sterols, glycerides and fatty acids. The data on their quantitative content is summarized in Table 1.

Physiologically active phospholipids (phosphatidylserine, phosphatidylethanolamine and phosphatidylcholine) participate in various metabolic processes and energy metabolism.

Phospholipids are a fundamental component of the structure of cell membranes and organelles. Their functional role is based on the control of cell membrane permeability. Maintains the functioning of cells, ie, ion exchange, respiration, biological oxidation, and affects the activity of enzymes in mitochondria. Phospholipid deficiency leads to impaired hepatic fat metabolism and consequent steatosis.

The fatty acid fraction of "Milife" contains physiologically active fatty acids which fall into the essential category. Of total fatty acids
Over 50% belong to linoleic acid, a precursor to prostaglandins that regulate hormonal activity in animals and humans.

Due to their known pharmacological properties, phospholipids and unsaturated fatty acids, like the known formulation Essentiale, regenerate damaged mitochondria and activate impaired enzyme systems. Enhances the detoxification function of the liver [MDMashkovsky: "Medicinal Agents", Medisina publisher, Moscow, Russia (1984), Vol. 2, 46 page].

High physiological activity is indicated by a one of the group D vitamins (D ₄₎ 22,23-dihydro-ergosterol. Its activity is cholecalciferol (vitamin D ₃ )
Analogous to that of, especially for growing organisms,
Facilitates calcium exchange and timely deposition of substances on growing bone [MDMashkovsky: "Medicinal Agents", Medisina publisher, Moscow, Russia (1984), p. 34].

Ubiquinone Q ₉ (ubiquinone) is a metabolite of an animal organism, while ubiquinone Q ₁₀ is a human metabolite.

Comparison tests with Yubinon or ubiquinone Q ₁₀ and vitamin E (active antioxidant) is anti-oxidation and anti-cytolytic activity of Yubinon or ubiquinone Q _10, it has been proven to be five times the vitamin E. Yubinon or ubiquinone Q ₁₀ are five times more active than vitamin E, a 1.5 times greater activity than lipoic acid, normalizing detoxification and secretory function of the liver damaged by carbon tetrachloride.

Yubinon or ubiquinone Q _10, the biological membranes, and animals that organisms and human, such as hepatocytes, is a generic component that can not alternate and when functioning cell metabolism as a whole. Its high hepatoprotective activity is the result of the inhibition of membrane pathogenic molecular mechanisms associated with enhanced oxidation of lipids to peroxide.

Known experimental data include ubiquinone and ubiquinone Q ₁₀
Provide evidence for the conclusion that it is characterized by antioxidant activity in toxically damaged liver tissue. Yubinon and ubiquinone Q ₁₀ may prevent cell lysis and necrosis of liver cells, normalizing the function of the secretion and detoxification of the liver affected by toxic hepatitis [LV
Vinogradov, EAOboln
ikova et al .: "Ubiquinones-Prospective hepatoprotective metabolites (Perspective Hepatopr)
otectors of Metabolic Tepe), 1st Russian National Conference on Humans and Pharmaceuticals, April 1992, Moscow, Russia, Paper No. 408].

Analysis of the composition of the lipid component of the formulation "Milife" can confirm its high physiological activity in vivo. The composition of the minerals of "Milife" is listed in Table 2 below.

From the data in Table 2, it can be seen that "Milife" contains large amounts of physiologically active elements.

Calcium in the body participates in the construction of bone tissue and the activation of blood clotting enzymes, reduces the excitability of some nervous system compartments, weakens the effects of toxins, increases resistance to infection, Increase electric potential.

Potassium normalizes intracellular pressure and activates many enzymes.

Phosphorus and its compounds function as constituents of bone tissue and fungi and participate in the energy production process.

Copper participates in the conversion of iron into a form that can be used for the synthesis of hemoglobin.

Manganese affects glycogen synthesis and participates in the normalization of carbohydrate metabolism.

Iron is taken up by hemoglobin and iron deficiency leads to a serious disease-anemia.

Thus, "Milife" contains a wide range of chemical elements, and the current concept regarding their role in the human body confirms the physiological activity of this formulation.

Table 3 lists the water-soluble vitamins contained in "Milife".

"Milife" mainly contains vitamins of group B, the total amount of which reaches 2.6-3.1%.

At present, the physiological effects of vitamins of group B are well studied. Plays an important part in the metabolic processes of proteins, fats and carbohydrates, increases the body's resistance to contagious diseases, due to their immunostimulatory effects, participates in the synthesis of prostaglandins, enzymes and coenzymes, Shows a strong protective effect. The "Miraifu", these vitamins, the or synthetic analogs, the form of artificially developed compositions (e.g. Methiovit (Mechiovito) Vitamin B ₁ hereinafter) over substantial advantages are natural complex Included in.

Biomedical Study of "Milife" The purpose of the biomedical study of "Milife" involved obtaining data to ensure the safety of its administration for therapeutic and prophylactic purposes.

Studies performed on three types of animals (rats, chickens, pigs) demonstrated that oral administration of "Milife" resulted in the following:-10, 25, 50 and 50% Under the conditions of the acute (30 days), subchronic (60 days) and chronic (1 year) experiments replacing 100% with "Milife", there are no negative effects;-spontaneous tumors and induced There is no carcinogenic or modifying effect on tumor development; no dominant lethal mutations in the sexual cells of the animal; no teratogenic and embryogenic effects in the 5 generation of animals; -No allergic effects;-No effect on heart, liver, kidney and spleen weight. -There is no effect on the morphological indicators of the blood of the animals, they remain within the limits of physiological norms.

Studies of the "Milife" formulation showed a relatively high activity of succinate dehydrogenase, and a relatively high glycogen content, in the liver of rats whose protein requirements were completely satisfied by the "Milife" formulation. Decreased fat content was also observed in rat liver where the protein was derived only from "Mylife".

Similar results were recorded in pigs kept according to established physiological norms.

 The study was performed on three groups of animals.

Group 1 ingested a conventional diet consisting of bone meal with protein components (30%).

The second group ate the same feed as the first group, but the protein component (30%) was "Milife" protein.

 The results of these experiments are shown in Table 4 below.

These data are evidence that "Milife" exerts a beneficial effect on lipid metabolism by substantially reducing the content of cholesterol and triglycerides.

Clinical study of the therapeutic effect of "Milife" in cases of dietary obesity (disorder of lipid metabolism) It is generally known that dietary obesity necessarily causes pathological conditions in various organ systems of the living body. It is.

In this regard, three groups of patients were given "milife": 1. Obese patients with metabolic disorders (hypercholesterolemia, arterial hypertension, diabetes with impaired glucose tolerance)-45, 2 .18 obese individuals with biliary conditions (chronic cholecystitis, cholelithiasis, bile duct dysmotility), 3. gastrointestinal tract conditions (gastric ulcer, erosive gastritis and duodenitis, duodenitis) Obesity cases with coexistence-12 patients.

1. A study of the effect of "Milife" on the dynamics of clinical biochemical indices in obese patients with metabolic disease. 45 patients in the observed group were diagnosed with obesity with hypercholesterolemia. Affected (Group 1-20 boys, Group 2-25 women).

Studies of conventional indicators of lipid metabolism, such as total cholesterose (TC), high-density lipoprotein (HDLP), triglyceride (TG) content, and atherogenic coefficient (CA) in serum A natural shift of the index was shown (Table 5). The analysis of indicators of the risk of developing ischemic heart disease (IHD) has generated special interest, because the overwhelming number of patients (33) in the analyzed group were under 40 years old It is.

Indicator HD for both the entire group of patients and individual men and women
The meaning of LP / TC is that they are within the high risk of developing this condition, that is, 17.6-18.7% compared to 20-25% of ordinary people
It was in that. The atherogenicity factor was reliably above the normal value (3.0 against normal value).
4.5 ± 0.4).

In 50% of the patients observed, there were two risk factors leading to the development of IHD (hypercholesterolemia in addition to hypertension). After one course of treatment, there was a marked decrease in lipid index in both the entire patient and in the men and women group.

Note: AP-arterial pressure TC-total cholesterol HDLP-high density lipoprotein HDLP / TC,%-ratio of HDLP to total cholesterol CA-atherogenicity factor TG-triglyceride The most notable changes are male, cholesterol levels. , A atherogenic index, a statistically significant decrease in triglycerides, and relatively high values of HDLP / TC were recorded.

The comparison between the obtained results and the control data
A more remarkable effect was revealed in the patients receiving. The effects of conventional treatments were less pronounced.

Initial examination of 15 patients showed a higher blood sugar content. After careful examination (glycemic curve with 75 g glucose load, glycemic index), 5 patients were diagnosed with type 2 diabetes and 10 patients with impaired glucose tolerance. In the group of patients receiving “Milife”, a statistically significant reduction in this factor was 7.54 ± 0.9 mmol compared to 6.32 ± 0.6 mmol in the control to 6.32 ± 0.6 mmol in the control.
From 5.3 mmol to 5.3 ± 0.48 mmol. The formulation was well tolerated in all patients. In 75% of the patients observed, starvation and a marked decline in appetite were noted. Weight loss is 11.9 ± 1kg for boys and 9.2 ± 2.1k for girls
g.

2. Effect of "Milife" on the dynamics of clinical and biological indices in patients with hepatic gallbladder pathology Eighteen patients under observation had various hepatic gall bladder pathologies. Dyskinetic dysfunction, such as hyperactive dyskinesia of the bile release duct, was found in 50% of cases. 50% of patients
Had a more pronounced organic pathology in the form of cholelithiasis confirmed by X-ray. At the time of hospitalization, all patients have a feeling of weight in the right intercostal region,
He also complained of paroxysmal pain after strong physical tension. These patients also suffered from various dyspepsia disorders such as bitterness and nausea following a fat meal. The patient received a conventional course of diet aimed at losing weight in combination with the administration of a bile releaser.

Gived "Milife" during the 21-day treatment cool. The tolerability to this formulation was good in all patients without exception. Appetite declined in 75% of cases.

 Table 6 shows the kinetics of the biochemical indicators.

From the above data, it can be concluded that patients in this group of conditions as a whole suffer from abnormalities characteristic of the entire group of obese individuals. Increased serum bilirubin levels due to biliary retention syndrome tended to decrease more significantly in the group of patients receiving the test formulation. Other findings include reduced appetite and hunger, and lipid levels (TC,
HDLP, TG), which leads to the conclusion that it is advantageous to administer the preparation to patients in this category.

Thus, "Milife" efficiently normalizes serum cholesterol and triglyceride levels in patients affected by cardiovascular disease, which progresses against a background of impaired lipid metabolism, thereby achieving ischemic heart disease. Risk rate can be reduced.

Study on the adaptation effect of "Milife" Tumor (cervical cancer, stage 2 and 3
Stage, kidney cancer, stage 3, lymphogranulomatosis)
The study was conducted in 15 year old tumor patients. The control group consisted of 15 individuals of the same age and tumor location. All patients received phototherapy at a dose of 2 to 4 Grays for a total of 40 to 60 Grays. Leukopenia was reduced in all patients receiving "Milife" compared to the control group (1,
600 + 370, P <0.05). Even in cases where the initial level of leukocytes is low (1.7 × 10 ^{8 to} 1.9 × 10 ⁸ ), 3.0 × 10 ^{8 in} 2 to 5 days
It could be raised to ~ 4.0 × 10 ⁸ , and sometimes even to 5.5 × 10 ⁸ . Although the use of other biostimulants could increase white blood cell counts,
Higher stability of the results has been demonstrated.

Increases in white blood cell counts were stable and often occurred on continuous irradiation, but this is very rare, especially without blood transfusions. A complete course of phototherapy could be administered without unplanned interruptions caused by leukopenia and blood transfusion.

The percentage of monocytes was high, but somewhat lower than that of the control group. That is, in 10 out of 15 cases, the monocyte standard is 3 to
Only 5% exceeded, and the remaining 5 and control patients had marked monocytosis (18-26%).

All patients showed reduced asthenia, improved appetite, sleep and mood.

Thus, "Milife" is an adaptive forming agent,
Studies have demonstrated the fact that it protects living organisms against damaging factors such as repeated irradiation.

The effects of "Milife" were also examined in patients with spontaneous leukopenia (15) and in patients with radiation-induced leukopenia (11). White blood cell counts were determined in patients who consumed "Milife" during 15, 30, and 60 days (Phase 1) and during the repeated courses 12 months later (Phase 2). The results are summarized in Table 7.

The data in Table 7 clearly demonstrate the kinetics of increased white blood cell count in patients affected by leukopenia of different etiologies.

Thus, the adaptive effect of "Milife" is indicated by increased non-specific tolerability of the organism in cases of leukopenia of different etiologies.

Study of the effect of "Milife" on the immune process A study was conducted on children aged 3-5 years with diagnosed hyperoxidation syndrome who live in ecologically disadvantaged areas. The total number of children examined was 77, 56 in the main group (1) and 21 in the control group (2).

Children in the main group were given "Milife" daily for 20-25 days.

 The control group received standard treatment in the hospital.

Both groups of children were carefully examined by pediatricians, neuropathologists, otolaryngologists, immunologists, microbiologists and other specialists.

Of the 56 children in the main group, 39 were found to be sensitized to visceral tissue, 21 for chronic gastritis, 18 for chronic colitis with symptoms of dysbacteriosis, 32 The name was diagnosed as having chronic cholecystitis and cholecystitis.

Apart from routine investigations (analysis of total blood counts, urine, feces, ECG), for anti-organ and anti-bacterial antibodies, the ultra-low levels of biological media and cells in the visible region spectrum caused by lipid structure Based on the glow, immunochemiluminescence was performed to test blood and saliva with a "Beta 2-analyzer".

The changes in autoimmune values produced by administration of "Milife" are listed in Table 8.

From Table 8, it can be concluded that the blood of both the main and control children accumulated large amounts of anti-organ antibodies and had significant autoimmunity to spleen, liver, small intestine, large intestine and stomach tissues. it can. Comparison of treatment results in the main and control groups shows that the reduction in autoimmunization efficiency in control children was relatively unnoticeable in the digestive tract and uncertainly reduced in spleen, lung and heart tissue There was found.

The prescription of "Milife" for children in the main group for 20-25 days,
As for the spleen, kidney, liver, stomach and intestine, they contributed to a reliable reduction of circulating autoimmune antibodies.

Therefore, it is feasible to prescribe "Milife" for cases of gastrointestinal disorders involving the autoimmune process and weakening of the body's defenses.

The effect of "Milife" on the microbiota of children in the same group was examined.

In 54.2% of children, perturbation of one or more indicators characteristic of skin microbiota, in 38.2%, relatively large number of microorganisms per unit of skin surface, in 36.8%, mannitol We focused on degrading bacteria and, at 44.5%, hemolytic bacteria that are not common in healthy children.

Disruption of the intestinal microbiota causes a substantial reduction in the inoculation rate of bifidobacteria; detection of coliforms with reduced fermentation activity; lactose-negative forms of Escherichia coli; and the appearance of large numbers of staphylococcal and streptococcal forms. Are characterized by altered normal relationships between various species of gut microbiota.

In 6.4% of the children, fecal inoculation showed the presence of active Candida fungi.

Therefore, this required a study of the potential for developing sensitization or bacterial infection of children by pathogenic autologous flora and treating them with "Mylife".

Sensitization to bacterial allergens was studied using the method of immunochemiluminescence.

The tests included measuring the accumulation of bacterial antibodies in blood and saliva against antigens of streptococci, staphylococci, E. coli, Proteus, enterococci, and Pseudomonas aeruginosa.

Studies of bacterial infection and sensitization have allowed the recording of immune responses to bacterial antigens. The most frequent response in children in both the main and control groups was sensitization to E. coli, staphylococci and proteus.

It has been found that changes in microbiota in children are often accompanied by enhanced antibody production against several types of bacteria.

Table 9 summarizes data obtained from studies of immune response to bacterial antigens in the main and control children.

The table concludes that administration of "Milife" to the main group of children contributed to a marked reduction in antibody levels against hemolytic staphylococci, streptococci, E. coli, enterococci and Pseudomonas aeruginosa.

Therefore, it is advantageous to prescribe "Milife" for children with symptoms of bacteriopathy that developed against the background of the impeded defense of the body.

Thus, administration of "Milife" to patients suffering from chronic diseases such as gallbladder cholangitis, gastritis, gastroduodeneitis, and colitis that often have the symptoms of dysbacteriosis that has developed on the background of hyperoxidation syndrome Reduced the autoimmune response to antigens primarily in the gastrointestinal tract and the concomitant desensitization to several bacterial allergens.

Practical applicability The fungal strain Fusarium sanbusinum VSB-917 is
It is used in the commercial manufacture of a formulation called "Milife".

The formulations claimed in the present invention are likely to prove valuable in the welfare business and can be used as:-disorders of lipid, cholesterol and glucose metabolism, especially in geriatric medicine (arteriosclerosis) Additional therapeutic agents in cases of ischemic heart disease, liver disorders, obesity, type 2 diabetes); prophylactic agents against physical illness and complications that occur in the background of radiation therapy in neoplastic patients; Adjuvants to administer during treatments for global health improvement or detoxification;-prophylactic preparations against ecologically unfavorable conditions or progressive diseases of people living or working in areas of intense radiation; Medicine for the treatment of dysbacteriosis;-treatment of vitamin deficiency;

With its autoimmune activity in mind, this formulation can be used for organ or tissue transplantation.

Description of the preparation of the "Milife" preparation The preparation of this preparation comprises the following steps:-Preparation and sterilization of the liquid nutrient medium-Preparation of the mycelium for inoculation of the fungus; Growth of the mycelium in the fermenter; Concentration, extraction, drying and packaging of the formulation.

The carbon sources required for hyphal growth are molasses, sugar, sugar, sucrose, glucose, alcohol and cognac industry waste (alcohol and cognac malt dregs), milk whey, root and green body juices. be able to. Sodium, potassium and phosphorus are made from ammonium nitrate and potassium acid phosphate.

The nutrient medium is sterilized in a continuous sterilizer at 125 ± 2 ° C. for 3 minutes. The inoculum mycelium for the fermenter is grown first on the solid nutrient medium in a glass container, then in a liquid nutrient medium in a shake flask and finally in the inoculator under the following conditions: temperature -25 ± 2 ℃ pH - 5.5~6.2 ventilation - 1m ³ / m ³ / min inoculum concentration of dry matter of mycelia in the medium, 9～12G /
It is considered ready when l is reached. This medium in an amount of 5 to 10% by weight serves as an inoculum, which is transferred under aseptic conditions into the fermentor.

The mycelium is cultured in the fermenter under the same conditions and in a nutrient medium of the same composition as in the inoculator. The culture method may be either a periodic discharge-addition type or a continuous type.

The next technical step is concentration of the medium in a vacuum filter. The product is then dehydrated in an infrared dryer under mild temperature conditions (up to 50 ° C.) where the biological activity of the components is retained.

 Drying time is 15-20 minutes.

 Finally, the formulation is granulated, pelletized and packaged.

The duration of storage of the formulation thus produced without loss of its activity is 24 months at room temperature (17-25 ° C.).

The following are examples of the composition of the "Milife" formulation claimed in the present invention.

Example 1. Fungal hyphae were grown in a 200-liter fermenter using a drain-addition culture in a nutrient medium of the following composition (% by weight): Sugar beet molasses 4 (2 in reduced form) Ammonium nitrate 0.3 Acidic potassium phosphate 0.2 Water Amount that totals 100 pH 5.5-5.8 Inoculum is 24 hours, with 14 hours as the duration of one culture cycle.
Introduced in a volume of 5% by volume at a culture temperature of ± 1 ° C. The content of the completely dried product was 20 g per liter of the medium.
The productivity of the process was 1.4 kg / m ³ / h.

The formulation obtained had the following composition (% by weight): total protein (N × 6.25) 60 carbohydrates 13 lipids 8 nucleic acids 3 inorganic substances 8 vitamins 2.6 water 5.4 Example 2 Using the “drain-add” method, The mycelium was grown in a 30 liter fermenter in a nutrient medium of the following composition (% by weight): Crude sugar 2 Ammonium nitrate 0.3 Potassium acid phosphate 0.2 Maize extract 0.05 Water A total amount of 100 pH 5.7-6.0 Inoculum 8% by weight, culture temperature 25 ± 1 ℃
Met. The duration of one cycle was 13 hours. Next, 92% of the medium was drained, fresh medium was added, and the culture was continued. This operation was repeated after 13 hours. The concentration of the completely dried product was 20.1 g / l.

The efficiency of the process was 1.5 kg / m ³ / h.

The formulation obtained had the following composition (% by weight): Total protein (N × 6.25) 63 Carbohydrates 10 Lipids 4 Nucleic acids 6 Inorganic substances 6 Vitamin 3.1 Water 7.9 Example 3 Proliferate mycelium as in Example 2 Let the inoculum get 10
It was introduced in an amount of% by volume. Culture temperature is 26 ± 1 ℃, medium pH
Was 6.0 to 6.2.

In this case, the duration of the culture was 12.5 hours and the accumulation of completely dried matter was 20.2 g / l.

The efficiency of the process was 1.6 kg / m ³ / h.

 The composition of the formulation (% by weight) was as follows: total protein (N x 6.25) 61.5 carbohydrates 11.5 lipids 6 nucleic acids 4.5 inorganic substances 7 vitamins 2.8 water 6.7

Continued on the front page (51) Int.Cl. ⁶ Identification code FI A61K 35/74 ABS A61K 35/74 ADVA ADV AGZA AGZ C12N 1/14 A C12N 1/14 A61K 9/14 L // (C12N 1/14 C12R 1:77) (56) References JP-A-62-26694 (JP, A) JP-A-1-16590 (JP, A) JP-A-1-273595 (JP, A) JP-A-5-170784 (JP , A) (58) Fields investigated (Int. Cl. ⁶ , DB name) A61K 35/74 A23L 1/28 C12N 1/14 C12R 1:77

Claims (4)

(57) [Claims] 1. The strain Fusarium sambucinum Fuckel varieties Bulai V
The use of SB-917 (deposited as the deposit code VKPM (F-169) of the union depository of the VNI Igentika Institute) to normalize cholesterol in patients with cardiovascular disease from lipid metabolism disorders And can induce trigseride in serum, reduce the risk of progression of ischemic heart disease, increase organ tolerance, and serve as an adaptation source for protection against radiation exposure, and in leukemia, a good increase in white blood cell count. Prevention of diseases due to metabolic disorders, indications and immune effects, which can reduce abnormal sublimation of the autoimmune process, mainly in the gastrointestinal tract, and can serve as a hyposensitizer for bacterial allergens Composition for treatment. 2. Total protein (N × 6.25) 60 to 63% by weight Carbohydrate 10 to 63% by weight Fat 4 to 8% by weight Nucleic acid 3 to 6% by weight Mineral substance 6 to 8% by weight Vitamin 2.6 to 3.1% by weight Water remaining Is produced by dehydrating a culture medium formed by deep-culturing a single culture of the fungus strain Funsarim sambucinum Fuckel, which has the composition described above, to prevent cardiovascular disease from lipid metabolism disorders. It can normalize cholesterol in affected patients, induce triglycerides in serum, reduce the risk of progression of ischemic heart disease, increase organ tolerance, and serve as an indication for protection against radiation exposure, In leukemia, it is good for increasing white blood cell count, reduces abnormal sublimation of the autoimmune process, and is mainly used in gastrointestinal tract wells and as a hyposensitivity drug against bacterial allergens One can be. A method for producing a medicament for preventing or treating a disease due to a metabolic disorder, an adaptogen, or an immune effect. 3. The method according to claim 2, wherein the composition is produced in the form of tablets, granules, or powder. 4. The product carrier is various foods.
The production method described in 1.