Application of a highly safe hypoglycemic natural product composition in preventing sudden death in diabetic patients and related natural products

Abstract

The present invention relates to the field of pharmaceutical technology, and in particular to an application of a hypoglycemic natural product composition in preventing sudden death in diabetic patients and related natural products. The composition comprises a mixture of EGCG and caffeine, wherein the mass ratio of EGCG to caffeine is 2-8:1, and the daily intake of EGCG by diabetic patients is less than 800 mg; the mass ratio range of EGCG to caffeine can effectively reduce blood sugar, and since caffeinated refreshing drinks such as Red Bull, coffee, and cola are common and easily available, excessive caffeine has a risk of inducing sudden death in diabetic patients, but sufficient EGCG can effectively prevent sudden death in diabetic patients; the natural product containing EGCG to caffeine in a mass ratio of 2-8:1 has higher acceptability to diabetic patients and can effectively implement the work of stabilizing blood sugar.

Description

Technical Field

[0001] The present invention relates to the technical field of natural product compositions, and in particular to an application of a highly safe hypoglycemic natural product composition in preventing sudden death in diabetic patients and related natural products. Background Art

[0002] Caffeine (CAF; CAS number: 58-08-2) is a central nervous system stimulant and a xanthine alkaloid compound. Caffeine is a typical adenosine inhibitor that has the functions of enhancing brain sensitivity, increasing nerve excitability, promoting the body's metabolism, and acting as a diuretic. Caffeine is widely found in energy drinks such as coffee, tea, cola, and Red Bull. Therefore, caffeine is also the most commonly used psychotropic drug in the world. According to the "Scientific Opinion on the Safety of Caffeine" issued by the European Food Safety Authority in 2015, for non-pregnant adults, habitual intake of up to 400 mg of caffeine per day will not cause safety problems. Pregnant women habitually consume up to 200 mg of caffeine per day will not cause safety problems for the fetus. A single or habitual intake of 200 mg of caffeine by lactating women will not cause safety problems for breastfeeding infants. Calculated based on an adult (60 kg), it is equivalent to a non-pregnant adult consuming 6.67 mg / kg of caffeine per day without causing safety problems. The conversion factor between humans and mice is 12.3, which is equivalent to a caffeine dose of 82 mg / kg / d for mice. However, experimental data show that the upper limit of caffeine tolerance for diabetic patients is lower than the above data, suggesting that if diabetic mice consume more than 82 mg / kg / d of caffeine in food, or if diabetic patients consume more than 6.67 mg / kg / d, there are potential safety risks. Since caffeinated drugs or foods are easily available, it is unrealistic to prevent diabetics from purchasing and consuming them. How to reduce the risk of sudden death in diabetics when consuming caffeine is a research topic that cannot be ignored.

[0003] The blood sugar lowering function of EGCG has been recognized, but drinkers often cannot uniformly obtain significant blood sugar lowering effects by simply supplementing EGCG with daily tea. This is because the ratios of EGCG and other nutrients in tea are different. If pure EGCG is used alone to lower blood sugar, the upper limit of EGCG for healthy people is 800 mg / d. If the upper limit is exceeded, there will be a potential risk of causing liver damage. Using a combination of EGCG and caffeine to lower blood sugar can greatly reduce the intake of EGCG and have a better blood sugar lowering effect. At the same time, if a natural product with a suitable blood sugar-lowering ratio of EGCG and caffeine is cultivated, it will help promote diabetic patients to form a daily habit of consuming a combination of EGCG and caffeine, and stabilize the blood sugar lowering effect.

[0004] The patent application with patent number 202111096562.7 discloses a method for extracting and applying compound catechins. It uses alcohol extraction and enzymatic removal of impurities technology to obtain a catechin product with good food safety and stability, a purity higher than 90%, and caffeine lower than 0.5%. It is used to prevent and treat neurodegenerative diseases, gout, cardiovascular and cerebrovascular diseases, antibacterial and antiviral, lowering blood lipids, blood pressure and blood sugar, anti-cancer and anti-oxidation, constipation, dental caries, weight loss, etc.; however, due to the broad anti-disease spectrum, the patent has a unsatisfactory blood sugar-lowering effect, and the risk of sudden death caused by caffeine is not taken into account. Summary of the invention

[0005] In view of the current situation that the existing technology has great side effects on diabetic patients and is not highly acceptable, the present invention provides an application of EGCG in the preparation of a drug for preventing sudden death in diabetic patients caused by excessive caffeine, which can effectively lower blood sugar, has no risk of sudden death, and has the advantage of low side effects; the present invention also provides a natural product related to the application of EGCG in the preparation of a drug for preventing sudden death in diabetic patients caused by excessive caffeine, which can effectively lower blood sugar in diabetic patients, has no risk of sudden death, and has the advantage of low side effects.

[0006] The present invention is achieved by the following technical solutions:

[0007] The invention discloses an application of EGCG in the preparation of a drug for preventing sudden death in diabetic patients caused by excessive caffeine and related natural products, comprising a mixture of EGCG and caffeine, wherein the mass ratio of EGCG to caffeine is 2 to 8:1, and the daily intake of EGCG by diabetic patients is less than 800 mg; the mixed mass ratio range of EGCG to caffeine can effectively reduce blood sugar, and since caffeinated refreshing drinks such as Red Bull, coffee, and cola are common and easily available, excessive caffeine has the risk of inducing sudden death in diabetic patients, and therefore sufficient EGCG can effectively prevent sudden death in diabetic patients.

[0008] Preferably, the mass ratio of EGCG to caffeine is 8:1.

[0009] Preferably, the diabetes is type 2 diabetes.

[0010] Preferably, drugs or natural products are also included.

[0011] A natural product containing the above-mentioned EGCG for preparing a drug for preventing sudden death in diabetic patients caused by excessive caffeine, wherein the natural product includes one or more of tea and berries; the industrial or agricultural derivative also includes a mixture of a biological product containing EGCG alone and a biological product containing caffeine alone; the natural product has higher acceptability to diabetic patients and can effectively implement the work of stabilizing blood sugar.

[0012] Beneficial effects of the present invention:

[0013] (1) The mass ratio of EGCG to caffeine is 2 to 8:1, which can effectively lower blood sugar in diabetic patients.

[0014] (2) The total daily intake of EGCG is within the safe range for diabetics, preventing liver damage and effectively preventing sudden death in diabetics caused by excessive caffeine intake.

[0015] (3) The effective mass ratio of EGCG to caffeine for lowering blood sugar is 2 to 8:1, which can be used to guide the production process of related products such as tea, and is conducive to promoting the production of new products that effectively assist in lowering blood sugar.

[0016] (4) The EGCG plus caffeine composition of the present invention is matched with a mixture of tea, berries and natural products, which is convenient and quick to use, has both easy acceptance and hypoglycemic effect, can effectively penetrate into the daily life of diabetic patients, and can assist prescription hypoglycemic drugs such as metformin in stabilizing blood sugar.

[0017] (5) For diabetic patients with mild symptoms, the EGCG plus caffeine composition of the present invention, in combination with a suitable ratio of tea, berries and a mixture of natural products, can partially replace prescription hypoglycemic drugs. Although the hypoglycemic effect is slightly weaker, the side effects of the drugs are significantly reduced. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Schematic diagram of the changes in fasting blood glucose levels in rats intervened with different doses of EGCG.

[0019] Figure 2 Schematic diagram of the changes in random blood glucose values ​​in rats intervened with different doses of EGCG.

[0020] Figure 3 Schematic diagram of the changes in the area under the glucose tolerance curve of rats intervened with different doses of EGCG.

[0021] Figure 4 Schematic diagram of the changes in biochemical indicators of rats treated with different doses of EGCG.

[0022] Figure 5 Schematic diagram of the changes in fasting blood glucose levels in mice treated with EGCG / CAF.

[0023] Figure 6 Schematic diagram of body weight changes in mice after CAF and EGCG / CAF intervention.

[0024] Figure 7 Schematic diagram of the survival rate curve of CAF / EGCG-intervention mice.

[0025] Figure 8 is the general chemical structure formula of EGCG and caffeine. DETAILED DESCRIPTION

[0026] In order to make the purpose, technical scheme and advantages of the present invention more clearly understood, the present invention is further described in detail in conjunction with the embodiments below; it should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention; in the embodiments, unless otherwise specified, the means used are conventional means in the art; the terms "comprising", "including" or any other variations thereof used herein are intended to cover non-exclusive inclusions; for example, a composition, step, method, product or device comprising the listed elements is not necessarily limited to those elements, but may include other elements not explicitly listed or elements inherent to such a composition, step, method, product or device; in addition, the technical features involved in each embodiment of the present invention described below may be combined with each other as long as they do not conflict with each other; the experimental raw materials used in the embodiments of the present invention and the comparative examples are all commercially available products.

[0027] Example 1.1

[0028] A blood sugar lowering pharmaceutical composition, comprising: 160 mg / kg·BW EGCG.

[0029] Example 1.2

[0030] A blood sugar lowering pharmaceutical composition, comprising: 120 mg / kg·BW EGCG.

[0031] Example 1.3

[0032] A blood sugar lowering pharmaceutical composition, comprising: 80 mg / kg·BW EGCG.

[0033] Example 1.4

[0034] A blood sugar lowering pharmaceutical composition, comprising: 40 mg / kg·BW EGCG.

[0035] Example 1.5

[0036] A blood sugar lowering pharmaceutical composition, comprising: 10 mg / kg·BW EGCG.

[0037] Comparative Example 1.1

[0038] A pharmaceutical composition, comprising: pure water.

[0039] Experiment 1 Materials

[0040] 1) Animals:

[0041] Sixteen healthy Wistar rats aged 7-8 weeks and 48 hyperglycemic GK rats aged 9-11 weeks were purchased from Changzhou Cavens Experimental Animal Co., Ltd. All animals were housed under standard conditions (24±1℃; 12 / 12 h light-dark cycle), and rats were free to eat and drink water. Wistar rats were fed a low-fat and low-sugar control diet, and GK rats were fed a 60% high-fat model diet. All animal experiments were conducted in accordance with the Guidelines for the Care and Use of Laboratory Animals of Hunan Agricultural University and approved by the Laboratory Animal Welfare and Ethics Committee of Hunan Agricultural University (Approval No.: 2021-2137; Changsha, Hunan, China). Before the start of the animal experiment, fasting blood glucose and random blood glucose values ​​were measured. When the fasting blood glucose value of GK rats was >11 or the random blood glucose value was >16.7; when the fasting blood glucose value of Wistar rats was <11 and the random blood glucose value was <16.7, they could be included in the experiment. In the experiment, different doses of EGCG were gavaged once a day for 4 consecutive weeks.

[0042] 2) Sample: EGCG (purity 98%) was provided by Hunan Sanfu Biotechnology Co., Ltd.

[0043] 3) Instruments: No. 16 gavage needle, sterile syringe, electronic scale, etc.

[0044] Experiment 1 Methods

[0045] After 2 weeks of feeding, 16 healthy Wistar rats were randomly divided into a blank group (NC) and an EGCG control group (NC-EGCG), and 48 hyperglycemic GK rats were randomly divided into a model group (MC), Example 1.5, Example 1.4, Example 1.3, Example 1.2, Example 1.1, a total of 8 groups, 8 rats in each group, that is, 10, 40, 80, 120, 160 mg / kg·BW EGCG were selected as intervention doses, and intervention was performed once a day. According to the conversion factor of 6.2 between rats and humans, this is equivalent to a daily intake of approximately 96.8 mg, 387.1 mg, 774.2 mg, 1161.3 mg, and 1548.4 mg EGCG in humans (60 kg).

[0046] 2) Methods for measuring fasting blood glucose, random blood glucose, glucose tolerance, glycosylated hemoglobin, and glycosylated serum protein

[0047] After fasting for 12 hours, the tail was cut and blood was collected, and the fasting blood glucose value was measured by Roche blood glucose meter. The tail was cut and blood was collected without fasting, and the random blood glucose value was measured by Roche blood glucose meter. After fasting for 12 hours, 2g / kg of anhydrous glucose was gavaged, and the blood glucose value of mice was measured at 0 minute, 30 minutes, 60 minutes, 90 minutes, and 120 minutes after gavage, respectively. The blood glucose value and detection time were recorded, and the area under the curve (AUC) was finally calculated. The glycosylated hemoglobin content was measured using the glycosylated hemoglobin kit of Wuhan Huamei Bioengineering Co., Ltd., and the glycosylated serum protein content was measured using the glycosylated serum protein kit of Nanjing Jiancheng Bioengineering Institute.

[0048] Example 2.1

[0049] A blood sugar lowering pharmaceutical composition comprises: 80 mg / kg·BW of caffeine and 160 mg / kg·BW of EGCG.

[0050] Example 2.2

[0051] A blood sugar lowering pharmaceutical composition comprises: 40 mg / kg·BW of caffeine and 160 mg / kg·BW of EGCG.

[0052] Example 2.3

[0053] A blood sugar lowering pharmaceutical composition comprises: 20 mg / kg·BW of caffeine and 160 mg / kg·BW of EGCG.

[0054] Comparative Example 2.1

[0055] A pharmaceutical composition comprising: 160 mg / kg·BW EGCG.

[0056] Experiment 2 Materials

[0057] 1) Animals:

[0058] Fifty 6-7 week old db / db male mice (transgenic hyperglycemic db / db mice) and 10 male m / m mice of the same age (standard control for db / db mice) were purchased from Jicui Yaokang. All animals were housed under standard conditions (24±1℃; 12 / 12 h light-dark cycle), and mice were free to eat and drink. All animal experiments were conducted in accordance with the Guidelines for the Care and Use of Laboratory Animals of Hunan Agricultural University and approved by the Laboratory Animal Welfare and Ethics Committee of Hunan Agricultural University (Approval No.: 2021-2137; Changsha, Hunan, China). Before the start of the animal experiment, the fasting blood glucose value was measured, and when the fasting blood glucose value was >11, the animal could be included in the experiment.

[0059] 2) Samples: Caffeine (purity 98%) was provided by the National Engineering Research Center for Utilization of Plant Functional Ingredients, and EGCG (purity 98%) was provided by Hunan Sanfu Biotechnology Co., Ltd.

[0060] 3) Instruments: No. 8 gavage needle, sterile syringe, electronic scale, etc.

[0061] Experiment 2 Methods

[0062] 1) Grouping and drug dosage selection

[0063] After 2 weeks of feeding, 50 db / db male mice and 10 male m / m mice aged 6-7 weeks were randomly divided into a blank group (Con), a model group (Mod), a 160 mg / kg EGCG group, i.e., comparative example 2.1, an EGCG / CAF group (160:20) group, i.e., example 2.3, an EGCG / CAF group (160:40) group, i.e., example 2.2, and an EGCG / CAF group (160:80) group, i.e., example 2.1, for a total of 6 groups, each with 10 mice. Mice were intragastrically administered with EGCG plus CAF, once a day. According to the conversion factor of 12.3 between mice and humans, Example 2.1 is equivalent to a daily intake of about 387.1 mg of caffeine and about 774.2 mg of EGCG for a human body (60 kg), which is within a reasonable range.

[0064] Example 3.1

[0065] A pharmaceutical composition for preventing sudden death of diabetic patients caused by caffeine, comprising: 80 mg / kg·BW of caffeine and 160 mg / kg·BW of EGCG.

[0066] Comparative Example 3.1

[0067] A CAF pharmaceutical composition comprises: 80 mg / kg·BW of caffeine.

[0068] Experiment 3 Materials

[0069] 1) Mice: 20 6-7 week old db / db male mice (transgenic diabetic db / db mice) were purchased from Jicui Yaokang. All animals were housed under standard conditions (24±1℃; 12 / 12 h light-dark cycle), and mice were free to eat and drink. All animal experiments were performed in accordance with the Guide for the Care and Use of Laboratory Animals of Hunan Agricultural University and approved by the Laboratory Animal Welfare and Ethics Committee of Hunan Agricultural University (approval number: 2021-2137; Changsha, Hunan, China).

[0070] 2) Samples: Caffeine (purity 98%) was provided by the National Engineering Research Center for Utilization of Plant Functional Ingredients, and EGCG (purity 98%) was provided by Hunan Sanfu Biotechnology Co., Ltd.

[0071] 3) Instruments: No. 8 gavage needle, sterile syringe, electronic scale, etc.

[0072] Experiment 3 Methods

[0073] 1) Grouping and drug dosage selection

[0074] After 2 weeks of feeding, 20 db / db male mice aged 6-7 weeks were randomly divided into a CAF group (Comparative Example 1) and a CAF+EGCG group (Example 1), with 2 groups of 10 mice in each group. In this experiment, 80 mg / kg·BW of caffeine and 160 mg / kg·BWEGCG were selected as the intervention doses of Example 1, and the intervention was performed once a day. According to the conversion factor of 12.3 between mice and humans, this is equivalent to a daily intake of about 387.1 mg of caffeine and about 774.2 mg of EGCG for a human body (60 kg), which is within a reasonable range.

[0075] Experimental Results

[0076] Experiment 1: The experimental results of the present invention showed that before EGCG intervention, there was no significant difference in fasting blood glucose between the NC group and the NC-EGCG group, and there was no significant difference in fasting blood glucose between the MC group and the different doses of EGCG groups ( Figure 1 A). After 4 weeks of EGCG intervention, there was no significant difference in fasting blood glucose between the NC group and the NC-EGCG group, indicating that 160 mg / kg EGCG intervention would not cause the side effect of hypoglycemia in normal rats; while different doses of EGCG intervention showed a significant effect of reducing the fasting blood glucose of hyperglycemic rats, and showed a dose-dependent effect, among which 160 mg / kg EGCG intervention had the best effect ( Figure 1 B).

[0077] The experimental results of the present invention showed that before EGCG intervention, there was no significant difference in random blood glucose values ​​between the NC group and the NC-EGCG group, and there was no significant difference in random blood glucose values ​​between the MC group and the different doses of EGCG groups ( Figure 2 A). After 4 weeks of EGCG intervention, there was no significant difference in random blood glucose values ​​between the NC group and the NC-EGCG group, indicating that 160 mg / kg EGCG intervention did not affect the random blood glucose values ​​of normal rats; while different doses of EGCG intervention showed the effect of reducing the random blood glucose values ​​of hyperglycemic rats (morning, afternoon, and evening), among which 160 mg / kg EGCG intervention had the best effect ( Figure 2 B).

[0078] The experimental results of the present invention showed that before EGCG intervention, there was no significant difference in the area under the glucose tolerance curve between the NC group and the NC-EGCG group, and there was no significant difference in the area under the glucose tolerance curve between the MC group and the different doses of EGCG groups ( Figure 2A). After 4 weeks of EGCG intervention, there was no significant difference in the area under the glucose tolerance curve between the NC group and the NC-EGCG group, indicating that 160 mg / kg EGCG intervention would not affect the area under the glucose tolerance curve of normal rats; while different doses of EGCG intervention showed a significant effect of reducing the area under the glucose tolerance curve of hyperglycemic rats, and showed a dose-dependent effect, among which 160 mg / kg EGCG intervention had the best effect ( Figure 3 B).

[0079] The experimental results of the present invention showed that before EGCG intervention, there were no significant differences in the levels of glycated serum protein and glycated hemoglobin between the NC group and the NC-EGCG group, and there were no significant differences in the levels of glycated serum protein and glycated hemoglobin between the MC group and the different doses of EGCG groups ( Figure 4 ). After 4 weeks of EGCG intervention, there was no significant difference in the glycated serum protein content and glycated hemoglobin content between the NC group and the NC-EGCG group, indicating that 160 mg / kg EGCG intervention would not affect the glycated serum protein content and glycated hemoglobin content of normal rats; while different doses of EGCG intervention showed a significant effect of reducing the glycated serum protein content and glycated hemoglobin content of hyperglycemic rats, among which 160 mg / kg EGCG intervention had the best effect ( Figure 4 ).

[0080] Experiment 2: The experimental results of the present invention showed that before EGCG / CAF intervention, there was no significant difference in fasting blood glucose values ​​between the MC group and the EGCG / CAF group with different ratios ( Figure 5 A, 5C). After 4 weeks of EGCG / CAF intervention, EGCG (160) intervention alone and EGCG / CAF intervention with different ratios showed significant effects on reducing fasting blood glucose levels in hyperglycemic mice, and the hypoglycemic effect of EGCG / CAF intervention with different ratios was significantly better than that of EGCG (160) intervention alone, indicating that EGCG and CAF have a significant synergistic hypoglycemic effect, among which the CAF+EGCG group (160∶20) had the best intervention effect ( Figure 5 A, 5B).

[0081] Experiment 3: EGCG intervention significantly inhibited the weight loss of diabetic mice caused by CAF ( Figure 6 In addition, within 2 weeks of CAF intervention, 6 diabetic mice died, with a mortality rate of 60%; while within 2 weeks of CAF+EGCG combined intervention, no diabetic mice died ( Figure 7 ), indicating that EGCG significantly alleviated the sudden death of diabetic mice caused by CAF. Adding EGCG simultaneously with CAF intake could effectively prevent the negative effects of excessive CAF-induced sudden death in diabetic mice, which is beneficial to the combined hypoglycemic effect of EGCG and CAF.

[0082] The above-mentioned embodiments only express several implementation modes of the present invention, and the description thereof is relatively specific and detailed, but it should not be understood as limiting the scope of the invention patent.

[0083] For ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention should be based on the attached claims.

Claims

1. An application of EGCG in the preparation of a drug for preventing sudden death in diabetic patients caused by excessive caffeine, characterized in that: The medicine comprises a mixture of EGCG and caffeine, the mass ratio of EGCG to caffeine is 2 to 8:1, and the daily intake of EGCG is less than 800 mg.

2. The use of EGCG according to claim 1 in the preparation of a drug for preventing sudden death in diabetic patients caused by excessive caffeine, characterized in that: The mass ratio of EGCG to caffeine is 8:

1.

3. Use of EGCG according to claim 1 or 2 in the preparation of a drug for preventing sudden death in diabetic patients caused by excessive caffeine, characterized in that: The diabetes is type 2 diabetes.

4. The use of EGCG according to claim 1 in the preparation of a drug for preventing sudden death in diabetic patients caused by excessive caffeine, characterized in that: The drug is a natural product.

5. The use of EGCG in the preparation of a drug for preventing sudden death in diabetic patients caused by excessive caffeine as claimed in claim 4, characterized in that: The natural products include one or more of tea and berries.

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