RU2161002C1 - Food, general-restorative, treatment-and-prophylactic product from shark tissues and method of its production - Google Patents

Abstract

FIELD: food industry. SUBSTANCE: product may be used as general-restorative nourishing additive designed for activization of immune system and prophylaxis of oncodiseases; product may be used for introduction in food products applied for improvement and prophylaxis of diseases of osteomuscular system and connective tissues. The proposed product is fermentative hydrolyzate from cartilaginous tissue of sharks of the following species: Lamna ditropis, Halaerurus buergeri, Galeocerdo cuvier, Sgualus acanthias, Somniosus antarcticus, Somniosus pacificus. The product contains free amino acids in the amount no less 20 mas.% including aromatic amino acids: tryptophan, tyrosine and phenylalanine which total content is no less 22%; diamino acids: lysine and agrinine, no less 19%; dicarboxylic amino acids - aspartic and glutamic ones, no less 17%; glycine, no less 6%; taurine, no less 1.5% from total quantity of amino acids; the product also includes hyaluronic acid in the amount of 2.1-5.5 mg/g of dry product. Method of product preparation involves homogenization of tissue, extraction of ground raw material and separation of final product. Extraction is conducted by 0.9-2.0-% solution of sodium chloride for 10-18 h at temperature 5-10 C. Then fermentative hydrolysis is carried out by proteolytic ferment preparations at temperature 30-45 C for 15-24 h. Inactivization of ferments is carried out by heating up to 70-80 C for 15-20 min, and separation of prepared product after inactivization is conducted through calico filter. EFFECT: broadened assortment of food treatment-and-prophylactic products with higher biological activity. 9 cl, 3 ex

Description

 The invention relates to the food industry and can be used as a general strengthening, improving, nutritional supplement intended for boosting the immune system and preventing cancer, as well as for introducing into food products used to improve the course and prevention of diseases of the musculoskeletal system, connective tissue.

 Known fortifying agents obtained from raw materials of animal origin: Pantocrine and Rantorin - extracts from antler sika deer, red deer, red deer and reindeer (Mashkovsky M.D. "Medicines", t. 1, M .: "Medicine", 1993, p. 165-166).

 Pantogem (TU 429-021-001-92), the active principle of which is the extraction of spotted deer from whole blood.

A wide range of therapeutic and prophylactic agents is known (RF patent N 2083218, MKI ⁶ A 61 K 35/78, 31/66), which includes pollen and DNA at a ratio of 60-95: 5-40, respectively.

 Known therapeutic and preventive balm "Vitagen" (TU 9169-010-20785204-94), containing 37 components of plant and animal origin, including ginseng, eleutherococcus, magnolia vine, preparations from kelp, salmon milk, mummy.

 These funds are used in the rehabilitation of patients after various diseases, a decrease in the functions of the central nervous system, and a decrease in muscle and mental performance.

 The well-known therapeutic and prophylactic product MIGI-K-LP (TU 15-16-28-94), which has non-specific immunomodulating properties, is an acid hydrolyzate from mussel meat, containing melanoidins with a molecular weight of up to 5000 and over 5000 with a quantitative ratio (mg / ml of solution), equal to 28-50: 2-8, having a pH of 5.0-5.8 containing 28-33 wt.% solids and containing 0.85-1.2% of the total weight of the product. This product is from mussel meat and the method for its production is protected by RF patent N 2017439, MKI 5 A 23 L 1/333, A 23 G 1/04. The mussel meat hydrolyzate contains essential amino acids, low molecular weight nitrogenous compounds, trace elements, free fatty acids, as well as a number of substances of a melanoid nature. MIGI-K-LP is recommended for people in contact with ionizing radiation, receiving chemotherapy, patients with chronic inflammatory processes, immunodeficiency, metabolic disorders, burns, for wound healing in the postoperative period.

 Known therapeutic and prophylactic product (RF patent N 2065707, IPC 6 A 23 L 1/305, 1/325), containing meat of warm-blooded animals, fish and invertebrates and / or waste from their processing and, as biologically active substances, peptide immunostimulants - cytomedines - mainly gangliin, thymalin, thymogen, T-activin, vilosen, in the following ratio of components, wt. %: - meat of warm-blooded animals, fish and invertebrates and / or waste from their processing - 99.99965 - 99.99950; cytomedines - 0.00035 - 0.0005.

 Known drug from the tissues of marine aquatic organisms "Orbitar" (RF patent 2053779, IPC 6 A 61 K 35/56) with a nitrogen content of amino groups of 0.8-1.0% of the total mass, with a ratio of free amino acids to peptide groups of 6: 7, dry matter mass of 25-35 wt.% and at a pH of 4.5-6.0, used to increase human resistance to adverse environmental factors.

 Known therapeutic, food preparation (RF patent 2075950, IPC 6 A 23 L 1/29, A 61 K 35/100), which contains adaptogens of plant and / or animal origin and / or detoxifiers in an amount of 0.5-97, 0 wt. % It also contains multivitamins, macro- and microelements. All components are used taking into account their functional and energy significance and in effective quantities.

 The closest technical solution for the technical nature of both the product and the method is the product and the method for its preparation described in patent application WO 9623512 A1, IPC 6 A 61 K 35/60, (10) 0267.15.008.97.

 The treatment-and-prophylactic product is an aqueous extract of shark cartilage and has molecules with a molecular weight of 0-500 kD - it has anti-angiogenic activity, anti-inflammatory, anticollagenolytic activity, as well as directed antitumor antiproliferative activity.

 A method of obtaining an extract of shark cartilage includes the following operations: homogenization of cartilage, water extraction, centrifuging the mixture to separate the extract, fractionating it to obtain a common extract with mol molecules. masses 0-500 kD.

 The disadvantage of the prototype is the limited range of biological activity.

 The problem solved by the invention is the expansion of the range of food treatment and prophylactic products, obtaining a product with a wider spectrum of biological activity due to the additional presence of free amino acids and amino sugars, hyaluronic acid. The essence of the invention lies in the fact that the food treatment and prophylactic fortifying product with immunostimulating and antioxidant activities is an enzymatic hydrolyzate from shark cartilage of the species Lamna ditropis, Halaerurus buergeri, Galeocerdo cuvier, Squalus acanthias, Somusius saricus amino acidus anticus an amount of at least 20% by weight of the dry preparation; including aromatic amino acids - tryptophan, tyrosine and phenylalanine, the amount of which is not less than 22% of the total number of amino acids, diamino acids - lysine and arginine - not less than 19% dicarboxylic amino acids - aspartic and glutamic acids not less than 17%, as well as glycine not less than 6% and taurine not less than 1.5%. In addition, the composition of the product includes 2.1-5.5 mg / g of hyaluronic acid, not more than 15% protein by weight of the dry preparation and 0.4-1.5 mg / g of free amino sugars by weight of the dry preparation.

The claimed food fortifying therapeutic and prophylactic product of their shark cartilage tissue is obtained in the following way: raw materials — fresh or dry shark cartilage — are crushed and extracted with 0.9-2.0% sodium chloride solution for 10-18 hours at 5-10 ^o C, enzymatic hydrolysis is carried out using proteolytic enzyme preparations at 30-45 ^o C for 15-24 hours, inactivation of enzymes is carried out by heating to 70-80 ^o C for 15-20 minutes, the subsequent isolation of the target product is carried out by filtration, drying is carried out by freeze-drying or sprayed tion process. Enzymatic hydrolysis is carried out using a proteolytic enzyme preparation isolated from pyloric appendages of Far Eastern salmon (pylorin) with an activity of 100-200 U / g in an amount of 1-4% of the mass of ground cartilage, or a proteolytic enzyme preparation isolated from king crab hepatopancreas with an activity of 150 -250 U / g in an amount of 0.5-3% of the mass of ground cartilage, or using megaterina with an activity of 200-400 U / g in an amount of 1-4% of the mass of ground cartilage.

 The known preparation of dried shark cartilage "Bio-Shark", used as an oncological prophylactic. The low efficacy of the agent can be explained by the limited ability of pancreatic proteases to hydrolyze the native tissue of shark cartilage, since the oncological prophylactic effect of the aforementioned agent is determined by low molecular weight components. The proteolytic enzymes used in the proposed solution make it possible to transfer low molecular weight peptides into a soluble and easily digestible state, increase the total content of amino acids, amino sugars, and hyaluronic acid in the final product, which is proteoglycan and has the ability to inhibit the growth of a cancerous tumor.

 The invention is illustrated by the following examples.

 Example 1. Food fortifying therapeutic product obtained from the cartilage of sharks of the species Squalus acanthias, is an enzymatic hydrolyzate and contains: free amino acids in an amount of 20% by weight of the dry preparation, including aromatic amino acids - tryptophan, tyrosine and phenylalanine, the amount which is 22% of the total number of amino acids, diamino acids - lysine and arginine - 19% dicarboxylic amino acids - asparagine and glutamic - 17%, as well as glycine 6% and taurine 1.5%, free amino sugar - 0.8 mg / g, protein - 1 0%, hyaluronic acid - 4.8 mg / g.

This product is prepared as follows: 1 kg of shredded cartilage sharks of the species Squalus acanthias, was extracted in 3.8 L of a 1.5% sodium chloride solution for 12 hours at 6 ^° C, then the extract was heated to 37 ^° C, the pH of the solution was adjusted to 8.0 and 30 g of pylorin with a proteolytic activity of 150 U / g, previously dissolved in 0.2 l of the same solution, were added, thoroughly mixed and kept for 16 h at the set temperature, stirring was carried out periodically. To inactivate the enzyme, the solution was heated to 80 ^° C and held for 15 minutes. The solution was filtered through a calico filter, the precipitate was discarded, and the filtrate was freeze-dried. The yield of the drug was 7%.

 The drug was tested in mice, which were injected intramuscularly with 3,4-benzopyrene in a dose of 2 mg / mouse to induce a tumor. The experimental group received the drug at a dose of 0.15-0.2 mg / mouse orally by drinking for 20 days before the introduction of 3,4-benzopyrene and then for 7 months. As a result of the experiment, it was found that in mice receiving the drug from cartilage of sharks, the development of an induced tumor was not observed in 12.5% of cases, while in the control group by the end of the experiment, the survival rate was 0%, and in the control group with the addition of the drug from cartilage sharks 31%

 Example 2. Food fortifying treatment and prophylactic product obtained from the cartilage of sharks of the species Lama ditropis, is an enzymatic hydrolyzate and contains free amino acids in the amount of 18% by weight of the dry preparation, including aromatic amino acids tryptophan, tyrosine and phenylalanine, the sum of which accounts for 20% of the total number of amino acids, diamino acids - lysine and arginine - 17%, dicarboxylic amino acids - aspartic and glutamic - 14%, as well as glycine 4.5%, taurine 1.2%, free amino sugar - 1.2 mg / g protein - 8.5% hyaluronic acid - 4.4 mg / g.

This product is prepared as follows: 1 kg of shredded cartilage of sharks of the species Lama ditropis was extracted in 3.8 L of 0.9% sodium chloride solution for 18 hours at 10 ^° C, then the extract was heated to 40 ^° C, the pH of the solution was adjusted to 8 and 15 g of a proteolytic preparation from hepatopancreas of king crab with a proteolytic activity of 220 U / g, previously dissolved in 0.2 L of the same solution, was added, thoroughly mixed and kept for 19 h at the set temperature, stirring was carried out periodically. To inactivate the enzyme, the solution was heated to 80 ^° C and held for 15 minutes. The solution was filtered through a calico filter, the precipitate was discarded, and the filtrate was freeze-dried. The yield of the drug was 8%.

 The drug was tested in mice, which were injected intramuscularly with 3,4-benzopyrene in a dose of 2 mg / mouse to induce a tumor. The experimental group received the drug at a dose of 0.15-0.2 mg / mouse orally by drinking for 20 days before the introduction of 3,4-benzopyrene and then for 7 months. As a result of the experiment, it was found that in mice receiving the drug from cartilage sharks, the development of an induced tumor was not observed in 12% of cases, while in the control group by the end of the experiment, survival was 0%, and in the control group with the addition of the drug from cartilage sharks - 30% .

 Example 3. Food fortifying therapeutic product obtained from the cartilage of sharks of the species Somniosus pacificus, is an enzymatic hydrolyzate and contains: free amino acids in the amount of 19% by weight of the dry preparation, including aromatic amino acids tryptophan, tyrosine and phenylalanine, the amount which is 18.5% of the total number of amino acids, diamino acids - lysine and arginine - 15%, dicarboxylic amino acids - asparagine and glutamine - 15%, as well as glycine 5% and taurine 1.0%, free amino sugar - 1.2 mg / g, protein - 8.8%; hyaluronic acid - 4.5 mg / g.

This product is prepared as follows: 1 kg of shredded cartilage of sharks of the species Somniosus pacificus, was extracted in 3.8 L of a 2.0% solution of sodium chloride for 10 hours at 8 ^° C, then the extract was heated to 45 ^° C, the pH of the solution was adjusted to 8 and 25 g of megaterina with a proteolytic activity of 280 U / g, previously dissolved in 0.2 l of the same solution, were added, thoroughly mixed and kept for 20 h at the set temperature, stirring was carried out periodically. To inactivate the enzyme, the solution was heated to 80 ^° C and held for 15 minutes. The solution was filtered through a calico filter, the precipitate was discarded, and the filtrate was freeze-dried. The yield of the drug was 6%.

 The drug was tested in mice, which were injected intramuscularly with 3,4-benzopyrene in a dose of 2 mg / mouse to induce a tumor. The experimental group received the drug at a dose of 0.15-0.2 mg / mouse orally by drinking for 20 days before the introduction of 3,4-benzopyrene and then for 7 months. As a result of the experiment, it was found that in mice receiving the drug from cartilage of sharks, the development of an induced tumor was observed in 12.5% of cases, while in the control group by the end of the experiment, survival was 0%, and in the control group with the addition of the drug from cartilage sharks - 31 %

Claims (9)

 1. Nutritional restorative treatment and prophylactic product from shark cartilage, characterized in that it is an enzymatic hydrolyzate from shark cartilage tissue of the species Lamna ditropis, Halaerurus buergeri, Galeocerdo cuvier, Sgualus acanthias, Somniosus antarcticus, Somniosus amino acids pomniosus somniosus somniosus and somniosus somniosus not less than 20 wt.%, including aromatic amino acids - tryptophan, tyrosine and phenylalanine, the amount of which is not less than 22%, diamino acids - lysine and arginine - not less than 19%, dicarboxylic amino acids - aspartic and glutamine - not less than 17%, and so e glycine at least 6% and taurine at least 1.5% of the total number of amino acids, but also contains hyaluronic acid in an amount of 2.1 - 5.5 mg / g dry formulation.  2. The product according to claim 1, characterized in that it contains not more than 15 wt.% Protein.  3. The product according to p. 1 or 2, characterized in that it contains 0.4 to 1.5 mg / g of free amino sugars by weight of the dry preparation. 4. A method of obtaining a food-enhancing therapeutic and prophylactic product from the cartilaginous tissue of sharks of the species Lamna ditropis, Halaerurus buergeri, Galeocerdo cuvier, Sgualus acanthias, Somniosus antarcticus, Somniosus pacificus, including tissue homogenization, extraction of the crushed raw material, which separates the final product, separation conduct a 0.9 - 2.0% solution of sodium chloride for 10 to 18 hours at 5 - 10 ^o C, then carry out enzymatic hydrolysis with proteolytic enzyme preparations at 30 - 45 ^o C for 15 to 24 hours, inactivation of the enzymes is carried out heating up to 70 - 80 ^o C for 15 to 20 minutes  5. The method according to claim 4, characterized in that the selection of the finished product after inactivation is carried out through a calico filter.  6. The method according to claim 4, characterized in that the enzymatic hydrolysis is carried out using a proteolytic enzyme preparation isolated from pyloric appendages of Far Eastern salmon (pylorin) with an activity of 100-200 U / g in an amount of 1 to 4% by weight of crushed cartilage.  7. The method according to claim 4, characterized in that the enzymatic hydrolysis is carried out using a proteolytic enzyme preparation isolated from king crab hepatopancreas with an activity of 150 - 250 U / g in an amount of 0.5 to 3% by weight of crushed cartilage.  8. The method according to claim 4, characterized in that the enzymatic hydrolysis is carried out using megaterina with an activity of 200 to 400 U / g in an amount of 1 to 4% by weight of crushed cartilage.  9. The method according to any one of claims 4 to 8, characterized in that the finished product is dried by freeze-drying or spraying.