阅读说明：本技术 一种辅助治疗糖尿病的核苷酸组合物及其制备方法和应用 (Nucleotide composition for adjuvant therapy of diabetes mellitus and preparation method and application thereof ) 是由 陈玉松 于 2021-07-27 设计创作，主要内容包括：本发明公开了一种辅助治疗糖尿病的核苷酸组合物及其制备方法和应用。该复配核苷酸是由AMP、CMP、GMP、UMP四种单核苷酸组成的混合物。其可以与维生素、矿物质等营养素或其他功能性物质(成分)搭配应用于药品、特医食品、保健品和功能性食品,可以改善机体糖耐量,促进胰岛素分泌,起到降血糖的作用。(The invention discloses a nucleotide composition for adjuvant therapy of diabetes, a preparation method and application thereof. The compound nucleotide is a mixture consisting of four mononucleotides, namely AMP, CMP, GMP and UMP. It can be used in combination with vitamins, minerals, etc. or other functional substances (components) in medicine, special medical food, health product and functional food, and has effects of improving glucose tolerance, promoting insulin secretion, and lowering blood sugar.)

1. The nucleotide composition for adjuvant therapy of diabetes is characterized by comprising compound nucleotides, wherein the weight ratio of the compound nucleotides is as follows: AMP 15.0-25.0%, CMP 15.0-45.0%, GMP 15.0-35.0%, UMP 15.0-30.0%.

2. The nucleotide composition of claim 1, wherein the weight ratio of the compound nucleotide is as follows: AMP 16.9%, CMP 43.2%, GMP 17.8%, UMP 22.1%, or AMP 24.3%, CMP 28.3%, GMP 19.3%, UMP 28.1%.

3. The nucleotide composition of claim 1 or 2, further comprising a nutrient or functional substance;

the weight ratio of the compound nucleotide to the nutrient is as follows: 8-232 parts of compound nucleotide, 0.16-0.9 part of vitamin A, 0.003-0.012 part of vitamin D, 3.18-7.15 parts of vitamin E and vitamin K₁0.018-0.04 parts of vitamin B₁0.34-0.76 part of vitamin B₂0.34-0.76 part of vitamin B₆0.34-0.76 part of vitamin B₁₂0.0005-0.0011 part, 0.84-1.89 part of nicotinic acid, 0.09-0.2 part of folic acid, 1.17-2.63 parts of pantothenic acid, 21.76-48.96 parts of vitamin C, 0.008-0.019 part of biotin, 334.71-522.99 parts of sodium, 451.86-706.04 parts of potassium, 0.18-2 parts of copper, 73.64-115.06 parts of magnesium, 3.3-9.2 parts of iron, 1.67-8.36 parts of zinc, 0.1-2.4 parts of manganese, 217.56-339.94 parts of calcium, 160.67-251.04 parts of phosphorus, 0.027-0.042 parts of iodine, 251.03-870.25 parts of chlorine and 0.1885029 parts of selenium, wherein the nutrient also comprises one or more of chromium, taurine and dietary fiber, 0.0067-0.2228 part of chromium, 32.2-80.4 parts of taurine and 3.50246 parts of dietary fiber;

the weight ratio of the compound nucleotide to the functional substance is as follows: 1-3 parts of compound nucleotide and 5-15 parts of functional substances, wherein the functional substances are one or more of resistant dextrin, taurine, linolenic acid microcapsule powder and nobiletin.

4. The method of preparing a nucleotide composition as claimed in any one of claims 1 to 3, wherein the ingredients are weighed according to the formulation and mixed well to form a finished product.

5. The method of claim 4, wherein the finished state comprises powders, granules, tablets, pills, oral liquids, capsules.

6. Use of the nucleotide composition of any one of claims 1-3 in the manufacture of a product for the adjuvant treatment of diabetes.

7. The use according to claim 6, wherein the product is a pharmaceutical product, a specialty medical food, a nutraceutical product, and a functional food.

Technical Field

The invention belongs to the field of medicine and health care, and particularly relates to a nucleotide composition for adjuvant therapy of diabetes, and a preparation method and application thereof.

Background

Diabetes is a metabolic disease characterized by hyperglycemia. Hyperglycemia is caused by a defect in insulin secretion or an impaired biological action or both. Insulin is a protein hormone secreted by islet beta cells in the pancreas stimulated by endogenous or exogenous substances such as glucose, lactose, ribose, arginine, glucagon, etc., is the only hormone in the body for reducing blood sugar, and promotes synthesis of glycogen, fat and protein.

Nucleotides are the basic building blocks of ribonucleic acid and deoxyribonucleic acid, and are precursors of nucleic acids synthesized in vivo. Nucleotides have important biological functions as they are distributed in the nucleus and cytoplasm of various organs, tissues and cells in the organism, and they participate in almost all biochemical reaction processes in the organism. The regulation of insulin secretion is closely related to the metabolism of various nucleotides in beta cells, and various nucleotide substances in cytoplasm influence insulin secretion simultaneously when insulin is secreted. Adenosine Triphosphate (ATP) plays a critical role in the regulation of insulin secretion by beta cells. Meanwhile, adenylic acid is also an important coenzyme, such as the components of coenzyme I (nicotinamide adenine dinucleotide, NAD +), coenzyme II (nicotinamide adenine dinucleotide phosphate, NADP +), Flavin Adenine Dinucleotide (FAD) and coenzyme A (CoA), and NAD + can promote the secretion of insulin, thereby supplementing the condition of insufficient insulin secretion of diabetics. In addition, the nucleotide has important functions of maintaining the immune function of the body, promoting the proliferation of immune cells and the secretion of cytokines. The supplement of nucleotide can well improve the immunity of the organism and improve the immunity of the organism. In the last two decades, exogenous nucleotides have been widely used in infant formula milk powder and health care products, and the safety of the exogenous nucleotides used in human bodies has been fully proved by scientific experiments.

CN 201880051788.5 discloses a composition and method of treatment using nicotinamide mononucleotide, wherein the composition comprising nicotinamide mononucleotide can be used for treating, ameliorating, reducing or reversing any disease or disorder involving NMN metabolism, such as diabetes, obesity, decreased insulin sensitivity, impaired glucose-stimulated insulin secretion, etc.

Disclosure of Invention

The invention aims to provide a nucleotide composition for adjuvant therapy of diabetes and a preparation method and application thereof. The compound nucleotide is a mixture of four mononucleotides, namely AMP, CMP, GMP and UMP according to a certain proportion, can be matched with nutrients or functional substances (components) such as vitamins, minerals and the like, and can play a role in adjuvant therapy and improvement of diabetes.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

the nucleotide composition for adjuvant therapy of diabetes comprises compound nucleotides composed of four mononucleotides of AMP, CMP, GMP and UMP, wherein the weight ratio of the compound nucleotides is as follows: AMP 15.0-25.0%, CMP 15.0-45.0%, GMP 15.0-35.0%, UMP 15.0-30.0%.

Further, the weight ratio of the compound nucleotide is as follows: AMP 16.9%, CMP 43.2%, GMP 17.8%, UMP 22.1%, or AMP 24.3%, CMP 28.3%, GMP 19.3%, UMP 28.1%.

Furthermore, the nucleotide composition also comprises nutrients or functional substances (components) such as vitamins, minerals and the like, and the compound nucleotide is matched with the nutrients or functional substances (components) such as the vitamins, the minerals and the like to prepare the composition to play a synergistic effect;

the proportion of the compound nucleotide to the nutrients such as vitamins and minerals is as follows: 8-232 parts of compound nucleotide, 0.16-0.9 part of vitamin A, 0.003-0.012 part of vitamin D, 3.18-7.15 parts of vitamin E and vitamin K₁0.018-0.04 parts of vitamin B₁0.34-0.76 part of vitamin B₂0.34-0.76 part of vitamin B₆0.34-0.76 part of vitamin B₁₂0.0005-0.0011 part, 0.84-1.89 part of nicotinic acid, 0.089-0.2 part of folic acid, 1.17-2.63 parts of pantothenic acid, 21.76-48.96 parts of vitamin C, 0.008-0.019 part of biotin, 334.71-522.99 parts of sodium, 451.86-706.04 parts of potassium, 0.18-2 parts of copper, 73.64-115.06 parts of magnesium, 3.3-9.2 parts of iron, 1.67-8.36 parts of zinc, 0.1-2.44 parts of manganese, 217.56-339.94 parts of calcium, 160.67-251.04 parts of phosphorus, 0.027-0.042 parts of iodine, 251.03-870.25 parts of chlorine and 0.1885029 parts of selenium, wherein the nutrient also comprises one or more of chromium, taurine and dietary fiber, 0.0067-0.2228 parts of chromium, 32.2-80.4 parts of taurine and 3.18846 parts of dietary fiber;

or the mixture ratio of the compound nucleotide and the functional substance (components) is as follows: 1-3 parts of compound nucleotide and 5-15 parts of functional substances (components), wherein the functional substances are one or more of Sichuan tangerine peel extract (nobiletin), linolenic acid microcapsule powder and resistant dextrin.

The invention also aims to provide a preparation method of the compound nucleotide composition, which is characterized in that the components are weighed according to a formula, uniformly mixed and prepared into a finished product according to a conventional process.

Further, the finished product state comprises powder, granules, tablets, pills, oral liquid and capsules.

The invention also aims to provide application of the nucleotide composition in preparing a product for assisting in treating diabetes. The finished product of the nucleotide composition can be directly added into medicines, special medical foods, health products and functional foods after being processed by conventional steps, and has the effect of assisting in treating diabetes.

The invention has the beneficial effects that: the invention provides a nucleotide composition for adjuvant therapy of diabetes, which comprises compound nucleotide consisting of four mononucleotides of AMP, CMP, GMP and UMP, or a mixture prepared by matching the compound nucleotide with nutrients or functional substances (components) such as vitamins, minerals and the like. After being ingested by human body, the nucleotide composition is decomposed into various free nucleotide substances under the action of various enzymes in the digestive tract of the human body, thereby promoting the secretion of insulin and enhancing the immunity of the human body, thereby achieving the effect of adjuvant therapy of diabetes. Nucleotide is food additive substance, and the safety of the nucleotide used in human body is well proved by scientific experiments.

Detailed Description

The present invention is further illustrated below by reference to specific examples, which are to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.

Example 1

The preparation method of the compound nucleotide comprises the following steps:

the compound nucleotide is prepared from 16.9 wt% of AMP, 43.2 wt% of CMP, 17.8 wt% of GMP and 22.1 wt% of UMP by conversion, weighing raw materials, and mixing with a mixer to obtain powder. All the operations are carried out in a clean area, and sampling is carried out according to the specification of a quality standard to carry out the inspection of each index.

Example 2

The preparation method of the compound nucleotide comprises the following steps:

the compound nucleotide is prepared by mixing 24.3 wt% of AMP, 28.3 wt% of CMP, 19.3 wt% of GMP and 28.1 wt% of UMP, weighing raw materials after conversion, and uniformly mixing by using a mixer to prepare powder. All the operations are carried out in a clean area, and sampling is carried out according to the specification of a quality standard to carry out the inspection of each index.

Example 3

The preparation method of the compound nucleotide composition comprises the following steps:

the compound nucleotide in this example is the compound nucleotide prepared in example 2.

According to the proportion of the compound nucleotide and nutrients such as nutrients and mineral substances, a certain amount of raw materials are weighed after conversion, and are uniformly mixed by a mixer to prepare powder. The operations are all operated in a clean area, and the finished product is sampled according to the specification of a quality standard to carry out the inspection of each index.

Example 4

The preparation method of the compound nucleotide composition comprises the following steps:

the compound nucleotide in this example is the compound nucleotide prepared in example 2.

According to the proportion of the compound nucleotide and nutrients such as nutrients, mineral substances and the like, a certain amount of raw materials are weighed after conversion, and are uniformly mixed by a mixer to prepare powder, tablets or capsules. The operations are all operated in a clean area, and the finished product is sampled according to the specification of a quality standard to carry out the inspection of each index.

Example 5

The preparation method of the compound nucleotide composition comprises the following steps:

the compound nucleotide in this example is the compound nucleotide prepared in example 1.

According to the proportion of the compound nucleotide and nutrients such as nutrients and mineral substances, a certain amount of raw materials are weighed after conversion, and are uniformly mixed by a mixer to prepare powder. The operations are all operated in a clean area, and the finished product is sampled according to the specification of a quality standard to carry out the inspection of each index.

Example 6

The preparation method of the compound nucleotide composition comprises the following steps:

weighing raw materials according to the proportion of 1.2g of compound nucleotide (the compound nucleotide prepared in the embodiment 2), 6.5g of resistant dextrin, 6g of crude linolenic acid microcapsule powder (linseed oil microcapsule powder) and 0.5g of taurine, uniformly mixing by using a mixer, and preparing into tablets. The operations are all operated in a clean area, and the finished product is sampled according to the specification of a quality standard to carry out the inspection of each index.

Example 7

The preparation method of the compound nucleotide composition comprises the following steps:

weighing raw materials according to the proportion of 2.4g of compound nucleotide (the compound nucleotide is prepared in example 2), 6.5g of resistant dextrin, 0.5 part of taurine and 0.034g of Sichuan tangerine peel extract (nobiletin), uniformly mixing the raw materials by a mixer, and encapsulating. The operations are all operated in a clean area, and the finished product is sampled according to the specification of a quality standard to carry out the inspection of each index.

Example 8 Effect test example

Materials and methods

1. Sample preparation: the nucleotide mixture samples obtained in examples 1 to 7 above.

2. Laboratory animal

SPF-level Kunming mouse, about 20g, experiment temperature: 24-26 ℃ and 65-70% of humidity.

3. Experimental dose: the nucleotide low dose group is 45mg/kgBW, the nucleotide medium dose group is 90mg/kgBW and the nucleotide high dose group is 135mg/kgBW, and distilled water is used for preparing the required concentration; the combination was administered at a dose of 0.1U/kg plus 45mg/kgBW in the nucleotide low dose group, 90mg/kgBW in the nucleotide medium dose group and 135mg/kgBW in the nucleotide high dose group.

4. Experimental methods

(1) Nucleotide mixture blood sugar reduction test on normal mice

Healthy male mice are selected, fasting is carried out for 12 hours without water prohibition, tail blood is taken for measuring the blood sugar value, the mice are randomly divided into two groups according to the blood sugar level, 1 control group and 1 nucleotide high-dose group, 10 mice in each group are provided, 135mg/kgBW is given to the nucleotide high-dose group, distilled water is given to the control group, continuous gavage is carried out for 30 days, fasting is carried out before, the fasting blood sugar value is measured, and the influence of a tested object on the fasting blood sugar of normal animals is observed.

(2) Nucleotide mixture blood sugar reduction test and glucose tolerance test for alloxan hyperglycemia model mice

A hyperglycemic model blood glucose reduction experiment is carried out on a mouse by utilizing a alloxan induced insulin resistance glucose metabolism disorder model (alloxan is an islet beta cell poison agent, can generate free radicals, selectively attacks islet beta cells, causes reduction of insulin secretion, leads to increase of blood glucose, and can affect other organs such as liver, so that functions of liver glycogen synthesis, glucose utilization and the like are inhibited, and thus experimental diabetes is caused). The fasting blood glucose and the glucose tolerance of the mice are used as judgment indexes.

Healthy male mice are selected, and experimental mice are divided into three groups, namely a blank control group (used for judging whether the model is successfully established), a control group and a test group. Firstly, a mouse diabetes model is constructed, the latter two groups of mice are fasted and are not forbidden to be watered for 12 hours, 45mg/kgBW tetraoxypyrimidine physiological saline solution is injected into the abdominal cavity for continuous days, and the model is constructed successfully after 5 days when the fasting blood glucose of the mice reaches more than 10 mmol/L. The constructed alloxan hyperglycemia model is divided into 1 model control group and 7 test groups, each test group is divided into three groups, namely a nucleotide low dose group, a nucleotide medium dose group and a nucleotide high dose group, and the number of the model control group and each group is 10 respectively. The mice in the test group were gazed daily with a certain dose of the physiological saline solution of the compound nucleotide of the formula of examples 1-7, which was 45mg/kgBW in the nucleotide low dose group, 90mg/kgBW in the nucleotide medium dose group, and 135mg/kgBW in the nucleotide high dose group, respectively. And (3) irrigating the other two groups (a blank control group and a model control group) with the same amount of physiological saline solution every day, after 30 days, taking tail blood to measure the fasting blood glucose and the insulin content of the mice, irrigating 2.0g/kgBW glucose solution for each group of mice, and taking tail blood to measure the fasting blood glucose of the mice after 0.5h and 2h respectively to perform a glucose tolerance experiment.

(3) Combined drug pair alloxan hyperglycemic model mouse hypoglycemic test

Healthy male mice are selected, and experimental mice are divided into four groups, namely a blank control group (used for judging whether a model is successfully established), a model control group, an insulin treatment group and a test group for combined administration of a nucleotide mixture and insulin. Firstly, a mouse diabetes model is constructed, the latter two groups of mice are fasted and are not forbidden to be watered for 12 hours, 45mg/kgBW tetraoxypyrimidine physiological saline solution is injected into the abdominal cavity for continuous days, and the model is constructed successfully after 5 days when the fasting blood glucose of the mice reaches more than 10 mmol/L. The constructed alloxan hyperglycemia model is divided into 1 model control group, 1 insulin treatment group and 4 test groups of nucleotide mixture and insulin combined drug group, each test group comprises three groups, namely a nucleotide low dose group, a nucleotide medium dose group and a nucleotide high dose group, and the model control group, the insulin treatment group and each group are respectively 10. The mice in the test group were gazed daily with a certain dose of the physiological saline solution of the compound nucleotide and the insulin according to the formulations of examples 2, 4, 6 and 7, which were 45mg/kgBW and insulin combined administration in the nucleotide low dose group, 90mg/kgBW and insulin combined administration in the nucleotide medium dose group and 135mg/kgBW and insulin combined administration in the nucleotide high dose group, respectively. And (3) performing intragastric administration on the blank control group and the model control group daily with the same amount of physiological saline solution, performing intragastric administration on the insulin treatment group daily with the same amount of insulin (only the insulin is added and the nucleotide mixture is not added) in the combined administration of the nucleotide mixture and the insulin, and after 30 days, taking tail blood to measure the fasting blood glucose and the insulin content of the mice.

Second, Experimental results of examples 1 to 7

1. Effect of nucleotide mixture on fasting plasma glucose in Normal mice

As can be seen from the results in Table 1, the nucleotide mixtures in examples 1-7 had no significant difference in fasting plasma glucose in normal mice compared with the control group, indicating that the nucleotide mixtures had no effect on fasting plasma glucose in normal mice.

TABLE 1 Effect of nucleotide mixtures on fasting plasma glucose in Normal mice in examples 1-7: (n＝10)

2. Influence of nucleotide mixture on fasting plasma glucose of alloxan hyperglycemic model mouse

As can be seen from the results in Table 2, compared with the blank control group, the fasting blood glucose value of the model control group is significantly increased (P is less than 0.05), which indicates that the hyperglycemia model is established; compared with a model control group, the nucleotide mixture of each dosage in examples 1 to 7 has the advantages that the fasting serum insulin content of the hyperglycemic model mouse is obviously increased, the blood sugar value is obviously reduced (P is less than 0.05), and the nucleotide mixture can promote the secretion of insulin and has the effect of reducing blood sugar.

TABLE 2 Effect of nucleotide mixtures on fasting plasma glucose in alloxan hyperglycemic model mice in examples 1-7: (n＝10)

Note:^#indicating that the difference is statistically significant compared to the blank control group;^*indicating that the difference is statistically significant compared to the model control group.

3. Effect of nucleotide mixture on glucose tolerance in alloxan hyperglycemic model mice

As can be seen from the results in Table 3, the blood glucose levels at 0.5h and 2h after glucose administration were significantly reduced (P < 0.05) by the nucleotide mixtures of examples 1 to 7 in comparison with the model control group, indicating that the nucleotide mixtures had the effect of increasing glucose tolerance in diabetic mice.

TABLE 3 Effect of nucleotide mixtures on fasting glucose tolerance in mice in examples 1-7 ((n＝10)

Note:^#indicating that the difference is statistically significant compared to the blank control group;^*indicating that the difference is statistically significant compared to the model control group.

4. Combined drug pair alloxan hyperglycemic model mouse hypoglycemic test

As can be seen from the results in Table 4, compared with the blank control group, the fasting blood glucose value of the model control group is significantly increased (P is less than 0.05), which indicates that the hyperglycemia model is established; compared with a model control group, the insulin treatment group remarkably increases the fasting insulin content of the hyperglycemic model mouse and remarkably reduces the blood sugar value (P is less than 0.05); compared with the insulin treatment group, the combination of the nucleotide mixture and the insulin in each dose in the examples 2, 4, 6 and 7 has the advantages that the fasting insulin content of the hyperglycemic model mouse is obviously increased, and the blood sugar value is reduced (P is less than 0.05); the nucleotide mixture can promote the secretion of insulin while the injected insulin reduces the blood sugar, and the nucleotide mixture and the insulin have the synergistic effect of reducing the blood sugar.

TABLE 4 Effect of the combination of nucleotide mixture + insulin on fasting plasma glucose in alloxan hyperglycemic model mice in examples 2, 4, 6, and 7: (n＝10)

Note:^#indicating that the difference is statistically significant compared to the blank control group;^*indicating that the difference is statistically significant compared to the model control group.

The experimental results show that: the fasting blood glucose value of the mice which take in the compound nucleotide is lower than that of the mice of the model control group, the serum insulin content is higher than that of the mice of the model control group, and the compound nucleotide can improve the sugar tolerance of the diabetic mice to a certain extent. The compound nucleotide and the mixture can improve the sugar tolerance of the organism, promote the insulin secretion and have the function of adjuvant therapy of diabetes.