RU2559066C1 - Biologically active food additive - Google Patents

Abstract

FIELD: food industry.

SUBSTANCE: biologically active additive contains 1-10% of low-molecular DNA of salmon fishes, 40-60% of glutamine amino acids, 0.01-0.05% of folic acid, 10-25% of amber acid, 20-30% of arabinogalactan complex forming agent. The biologically active additive is produced by way of the said components mixture mechanochemical treatment in the following mode: the weight ratio of the mixture treated to grinding balls is equal to 1/10; the drive rotation rate is equal to 300-500 rpm; the mechanical treatment time is equal to 5-15 minutes.

EFFECT: invention allows to obtain a biologically active additive acting directly on nucleic acids and proteins synthesis, improving tissue regeneration, recovering the immune system, promoting enhancement of physical performance and stamina.

2 cl, 5 tbl, 4 ex

Description

The invention relates to the food industry, it allows you to get a dietary product, which is a source of nucleic and other organic acids, which has a directed effect on the synthesis of nucleic acids and proteins, improve tissue regeneration, restore immunity, helps to increase physical activity and endurance.

Known biologically active additive (BAA) to the food "Biostim". BAA is obtained from the testes of cattle, contains nucleic acids (of which about 98% is DNA, 2% is RNA), is produced in tablets of 0.2 g. “Biostim” stimulates the function of the blood-forming organs, increases peripheral blood counts to normal (hemoglobin, red blood cells, white blood cells), reduced for various reasons (blood loss, adverse environmental factors, including ionizing radiation), stimulates non-specific factors of the body's defense, in particular, the phagocytic activity of neutrophils. Biostim is used for prophylaxis and in the complex treatment of anemia, for rehabilitation after diseases, injuries, surgeries, etc. (L.N. Fedyanina, N.N. Besednova, L.M. Epstein, T.K. Kalenik, Yu.G. Blinov // Pacific Medical Journal, 2007, No. 4, p. 9-12).

It is known that when dissolved in water, nucleic acids swell and form viscous solutions of the colloidal type. The disadvantage of this drug is the low efficiency associated with the low solubility of oligonucleotides in water, and low bioavailability of the drug. As a result, effective doses of nucleic acid based enteric preparations are about 500 mg / day.

The well-known drug "Derinat", which is a highly purified sodium deoxyribonucleate (Sodium deoxyribonucleate). The drug has a wide spectrum of action, is recommended for the following diseases: myelodepression and cytostatic resistance in cancer patients, acute pharyngeal syndrome, gastric and duodenal ulcer, gastroduodenitis, IHD, cardiovascular failure, chronic ischemic disease of the lower extremities II and III stage prostatitis, vaginitis, endometritis, infertility and impotence caused by chronic infections, chronic obstructive bronchitis (Chernov V.N., Sharkovskaya T.E. of the drug Derinat in the treatment of patients with trophic ulcers of the lower extremities // Biomedicine, 2006, No. 3, v. 1, pp. 87-88).

The disadvantage of the drug is that it is used in the form of an injection. The course of treatment provides from 5 to 10 intramuscular injections. Hypersensitivity, hyperthermia, allergic reactions are possible.

Disclosure of invention

The objective of the invention was the creation of a composition based on nucleic acids with high efficiency, but devoid of side effects. To satisfy these requirements, the composition must have high solubility, and new components must be introduced into the composition that improve the assimilation of oligonucleotides and enhance the action of nucleic acids.

The proposed biologically active food supplement is a composition containing DNA, glutamine, folic acid, succinic acid and arabinogalactan in the following ratio of components: low molecular weight salmon fish DNA - 1-10%, glutamine amino acid - 40-60%, folic acid - 0, 01-0.05%, succinic acid - 10-25%, complexing agent arabinogalactan - 20-30%.

All components of the mixture are used in the form of a dry powder.

In order to create water-soluble complexes, mechanochemical processing of the mixture is carried out in the following mode:

loading of the processed material in relation to the loading of grinding bodies - 1:10 (1 mass part of the processed mixture and 10 mass parts of grinding balls);

- drive rotation speed - 300-500 rpm;

- machining time - 5-15 minutes

The resulting mixture has the form of a fine gray powder with a particle size of 0.01-0.001 mm, is readily soluble in water, stable during storage.

Comparison of the quality indicators of the compositions obtained with different durations of mechanochemical processing (optimized parameter) showed that with a duration of more than 15 minutes, the chemical decomposition of nucleic acids and arabinogalactan occurs, as a result of which their content decreases by less than 80% of the theoretical value. When the duration of the mechanochemical process is less than 5 minutes, the solubility of the treated mixture does not significantly differ from the solubility in water of a simple mixture of arabinogalactan and DNA.

Our composition differs from the above analogues in that, in addition to nucleic acids, it includes glutamine, folic and succinic acids, which have a synergistic effect on the synthesis of nucleic acids and protein, as well as tissue respiration and energy metabolism, which is reflected in accelerated regeneration and restoration of cellular composition (example 5).

The proposed composition differs from analogues also in that it contains arabinogalactan, which, as a result of mechanochemical activation, forms water-soluble complexes with hydrophobic compounds of the general formula R2-R1-R2, where:

R1 - hydrophobic compounds (nucleic acids, glutamine, folic acid),

R2 - complexed due to van der Waals forces with hydrophobic compounds arabinogalactan.

The proposed composition in comparison with enteric analogues has a higher solubility (example 1) and efficiency (examples 3, 4, 5), and compared with injection analogues, it is highly safe. It has a directed effect on the synthesis of nucleic acids and proteins, improves tissue regeneration, restores immunity, helps increase physical activity and endurance.

Examples

Example 1. A mixture of components (low molecular weight DNA of salmon fish, glutamine, folic acid, succinic acid, arabinogalactan) is subjected to mechanochemical processing in the following mode:

- loading of the processed material - 0.2 kg;

- loading grinding media - steel balls with a diameter of 12 mm - 2 kg;

- drive rotation speed - 500 rpm;

- machining time - 7 minutes

After processing in this mode, the final size of the obtained fraction was 0.01-0.001 mm, the solubility in water of the resulting composition at room temperature was 96%.

Example 2. To obtain the claimed composition, the ingredients are mixed in the following ratio, wt. %: low molecular weight DNA of salmon fish (5%), amino acid glutamine (50%), folic acid (0.05%), succinic acid (20%), arabinogalactan complexing agent (up to 100%), after which the mixture is subjected to mechanochemical treatment.

Example 3. The study of the effect of the proposed composition on the restoration of protein synthesis and metabolism of nucleic acids in patients with nutritional deficiency.

Criteria for inclusion of patients in the study: patients of both sexes with cicatricial narrowing of the esophagus or achalasia of the esophagus, in need of diet correction in connection with a decrease in body weight; age from 18 to 45 years.

Clinical and laboratory methods were used to assess the nutritional status (NS): anamnestic data, height, body weight, rate of weight loss (the faster the patient loses body weight, the more pronounced the degree of nutritional deficiency), the calculation of ideal body weight (IDMT) using the Lorentz formula (X-100- (X-150) / 2, where X is the numerical value of growth taken in centimeters), calculation of the Ketle index (Ketle index (weight / height) = body weight (kg) / squared patient height in m) , body mass deficiency (DMT) from idmt. The data were taken to determine the absolute number of lymphocytes (ALF), the plasma content of total protein (OB), albumin (Alb), transferrin (Tr). Nucleic acid metabolism (NK) was evaluated by the level of DNA in serum.

In all patients, changes in nutritional status were obvious due to a lack of body weight - 9.6% ± 1.4.

Initial serum transferrin and DNA concentrations were low in all patients. In particular, the average Tr and DNA levels were in the range 1.3-1.7 g / L ± 0.1-0.07 g / L and 0.8-0.9 ng / ml ± 0.06-0. 1 ng / ml, respectively.

Two groups of patients were formed. The distribution of the groups was carried out blind randomly.

The first group consisted of patients (n = 11) with a traditional management technique: the patients ate normal food through a gastrostomy or an awakened esophagus. In the second sample (n = 12), patients also fed through a gastrostomy or through an awakened esophagus, and the claimed composition was included in the usual diet for 7-10 days. The dose of DNA was 50 mg / day, the total weight of the composition 2 g / day.

Thus, the Ketle index, total blood protein, and serum albumin concentration in blood serum were uninformative in these patients. According to modern guidelines, the levels of OB and Alb are indeed not an absolute guide in assessing nutritional status, primarily due to the lability of the concentration in the blood serum and dependence on the water balance. More informative are the concentrations of Transferrin and serum DNA. In patients of both groups there was a Transferrin deficiency, indicating severe nutritional deficiency (LV) (1.37 ± 0.06-1.4 ± 0.15). 7 days after the start of nutrition, the level of Transferrin increased in the first group to 1.81 ± 0.15, however, this increase is statistically unreliable. Moreover, in the 2nd group, against the background of the introduction of the claimed composition, the concentration of Transferrin was significantly higher by the 7th day both in comparison with the original numbers and compared with similar values in the 1st group and amounted to 2.6 ± 0.1 (p <0 , 05).

To assess the exchange of nucleic acids in patients of the first and second groups (a total of 23 patients from 20 to 65 years old male and female with a diagnosis of esophageal achalasia or cicatricial narrowing of the esophagus), we determined the concentration of blood serum DNA at all stages of the study (Table 2).

When analyzing the results in patients of the first group, a deep deficiency of DNA content was recorded (for comparison, in healthy donors, normal values of blood serum DNA are 10-60 ng / ml) in blood serum during all 7 days of observation. However, in the second group, after the appointment in the standard scheme of nutritional support in the form of the claimed composition, the DNA content in the blood serum increased tenfold (Table 2). This is due to the dependence of the synthesis of nucleic acids on the consumption of glutamine, folic acid and low molecular weight oligonucleotides.

Thus, when the claimed composition was included in the nutritional support of patients with nutritional deficiency, a rapid reliable restoration of the levels of Transferrin and serum DNA was noted, which are the main criterion for the effectiveness of protein metabolism and nucleic acid metabolism. The claimed composition has a reliable anabolic effect even in patients with initial impairment of nucleic acid metabolism.

Example 4. Assessment of physical performance using the Harvard step test.

After the completion of dosed physical activity, the subject is resting while sitting. Starting from the 2nd minute, his heart rate is calculated 3 times over 30-second time intervals: from the 60th to the 90th, from the 120th to the 150th and from the 180th to the 210th second of the recovery period. The values of these three counts are added up and multiplied by 2 (conversion from beats / 30 s to beats / min). Test results are expressed in arbitrary units in the form of a Harvard step test index (IGST), the value of which is calculated from the equation:

IGST = T (100 / (f2 + f3 + f4) × 2,

where Τ is the actual execution time of physical activity in seconds; f2, f3, f4 - the sum of heart rate for the first 30 from each (starting from the 2nd) minute of the recovery period.

Evaluation of test results. The value of IGST characterizes the speed of recovery processes after intense physical exertion and is evaluated on a scale. The faster the heart rate is restored after the step test, the lower the value of f2 + f3 + f4 and, therefore, the higher the IGST. Assessment of physical performance based on the results of the Harvard step test (Karpman V.L. et al. // Testing in sports medicine. M.: FiS, 1988. 120 pp.) In IGST, ed .:

less than 55 is a bad result,

55-64 - below average,

65-79 - the average result,

80-89 is good

90 and more is an excellent result.

The experimental group included 9 males aged 25-40 years. The claimed composition was taken at a dose of 2000 mg (in terms of DNA - 50 mg) in the morning, 15 minutes before meals 1 time per day daily throughout the study period.

The comparison group included 10 males aged 25-40 years. The composition of DNA + glutamine + folic acid + succinic acid + arabinogalactan in similar weight ratios, but not subjected to mechanochemical effects, was also taken at a dose of 2000 mg (in terms of DNA - 50 mg) in the morning, 15 minutes before meals, 1 time per day.

The results are shown in table. 3.

Thus, a simple mixture of components did not lead to a significant increase in endurance / performance. The use of the claimed composition in the indicated dose significantly increased working capacity / endurance 30 days after the start of administration.

Example 5. The effect of the claimed composition on the restoration of bone marrow and the level of lymphocytes in mice after exposure to a cytostatic agent.

Myelosuppression in male CBA / CaLac mice (body weight 20-25 g) was carried out by a single injection of fluorouracil (5-ftoruracil) at a dose of 150 mg / kg body weight.

The following study groups were formed:

- intact animals (7 individuals),

- control group of animals (7 individuals). After exposure to fluorouracil, physiological saline was injected,

- experimental group 1 (7 individuals): animals after a single exposure to fluorouracil were fed once daily with a solution containing 30 mg / kg of low molecular weight DNA of salmon fish,

- experimental group 2 (7 individuals): animals, after a single exposure to fluorouracil daily, throughout the entire observation period, were watered once a day with a solution containing a mixture of the following components without mechanochemical treatment of the mixture: low molecular weight salmon fish DNA - 2.5%, glutamine amino acid 57.97%, folic acid 0.03%, succinic acid 14%, arabinogalactan 25.5%. The total dose of DNA is 30 mg / kg,

- experimental group 3 (7 individuals): animals after a single exposure to fluorouracil were fed daily with a solution containing the mechanochemical treatment according to example 1 mixture: low molecular weight salmon DNA - 2.5%, amino acid glutamine 57.97%, folic acid 0.03%, succinic acid 14%, arabinogalactan 25.5%. The total dose of DNA is 30 mg / kg.

The lymphocyte count in the peripheral blood and bone marrow was evaluated. The results are shown in table. 4 and tab. 5.

From the table. Figures 4 and 5 show that low molecular weight DNA has a positive effect on the rate of bone marrow recovery and the level of peripheral blood lymphocytes in mice. The addition of the organic acids glutamine, folic acid and succinic acid potentiates this action of DNA. The complex, subjected to mechanochemical treatment, had the most pronounced effect. A significant myelostimulating effect in this group was noted already on the 3rd day after the administration of 5-fluorouracil. On day 14, hypercorrection with lymphocytosis was noted, while in the control group the level of lymphocytes reached only 52% of the initial values. By 14 days in group 5, a significant difference was achieved compared with all other experimental groups. Thus, it has been proved that low molecular weight DNA is most effective in combination with arabinogalactan, as well as with a synergistic effect of glutamine, folic acid and succinic acid after mechanochemical treatment of the mixture.

Claims (2)

1. Dietary supplement (BAA) for food, including DNA, characterized in that it additionally contains glutamine, folic acid, succinic acid and arabinogalactan in the following ratio of components: low molecular weight salmon fish DNA - 1-10%, glutamine amino acid - 40-60 %, folic acid - 0.01-0.05%, succinic acid - 10-25%, complexing agent arabinogalactan - 20-30%, and dietary supplements are obtained by mechanochemical processing of a mixture of the above components in the following mode: the ratio of the mass of the processed mixture of components and grinding media arov is 1/10, the rotation speed of the drive is 300-500 rpm; machining time - 5-15 minutes 2. Supplements for food under item 1, characterized in that it is a fine powder with a particle size of 0.01-0.001 mm