Application of antifebrile dichroa extract and derivative thereof in preparation of medicine for treating diabetes

Abstract

The invention provides application of antifebrile dichroa extract and derivatives thereof in preparation of drugs for diabetes, and relates to the technical field of biological medicines. In the application, the antifebrile dichroa extract is specifically antifebrile B, and a derivative of the antifebrile dichroa extract is halofuginone. Experiments prove that the antifebrile dichroa extract and derivatives thereof (halofuginone and febrile B) can effectively regulate blood sugar, reduce polydipsia and polyphagia symptoms of experimental animals, reduce body weight, increase sugar tolerance of the experimental animals, improve insulin resistance of the experimental animals, prevent complications such as cardiovascular diseases, neuropathy and nephropathy caused by diabetes, improve the life quality of patients, and prevent diabetes mellitus from being damaged by diabetes mellitus. And the long-term health risk is reduced.

Description

Technical Field

[0001] This application relates to the field of biomedical technology, and more specifically, to the use of Changshan extract and its derivatives in the preparation of drugs for diabetes. Background Technology

[0003] Currently, a wide variety of drugs are used clinically to treat diabetes, including insulin and its analogues, oral hypoglycemic agents such as metformin, sulfonylureas, and thiazolidinediones. However, these drugs have many limitations in treatment. For example, insulin requires injection, which is inconvenient for patients, and improper use can easily lead to adverse reactions such as hypoglycemia; while oral hypoglycemic agents are relatively convenient to use, long-term use may cause side effects such as gastrointestinal discomfort, liver and kidney damage, and weight gain, and some patients may even experience secondary drug failure. Therefore, developing safe, effective, and novel antidiabetic drugs has significant clinical and social value.

[0004] Changshan (Dichroa febrifuga) is a traditional Chinese medicine with a long history of medicinal use in my country. Modern research, utilizing advanced technologies, has conducted in-depth analyses of various active components of Changshan, such as chondroxyfen and chondroxybenzyl. Chondroxyfen exhibits diverse biological activities, including anticoccidia and antimalarial effects, while chondroxybenzyl also shows antimalarial and emetic effects. With ongoing research, the scope of research on Changshan's active components continues to expand, gradually extending from traditional medicinal efficacy to more potential applications. Further exploration of the other biological activities and mechanisms of action of chondroxyfen and chondroxybenzyl is expected to open new pathways for innovative drug development, bringing new hope for solving the treatment challenges of complex diseases such as diabetes. Summary of the Invention

[0005] The purpose of this application is to provide the application of Changshan extract in the preparation of drugs for diabetes, and to expand the application of Changshan extract (Changshan B) and its derivatives (Changshan ketone) in the field of diabetes.

[0006] To solve the above-mentioned technical problems, the technical solution adopted in this application is as follows: This application provides the use of styrax extract styrax ethylsin and its derivative styraxone in the preparation of a drug for diabetes.

[0007] The experiments conducted in this application revealed that the hypoglycemic effects of chrysanthemum ketone and chrysanthemum ethyl are the result of multi-factor regulation. From the perspective of blood glucose regulation, chrysanthemum ketone and chrysanthemum ethyl significantly reduced fasting and random blood glucose levels in BKS-db / db diabetic model mice. Regarding the improvement of metabolic symptoms, they reduced polydipsia and polyphagia in diabetic mice, decreasing food intake and water consumption. Simultaneously, they enhanced glucose tolerance, improved insulin resistance, increased glucose tolerance in diabetic mice, and promoted glucose utilization and metabolism. Furthermore, the study found that both can inhibit hepatic gluconeogenesis and reduce endogenous glucose production by reducing the level of p-CREB / CREB protein in the liver of BKS-db / db mice. Insufficient insulin secretion or pancreatic dysfunction is a significant contributing factor to diabetes. Chrysanthemum ketone and chrysanthemum ethyl can improve pancreatic tissue damage, promote insulin secretion, directly supplement insulin levels, enhance glucose uptake and metabolism, and achieve comprehensive regulation of diabetes.

[0008] Compared with the prior art, the embodiments of this application have at least the following advantages or beneficial effects: This application demonstrates through experiments that Changshan extract and its derivatives (Changshan ketone, Changshan B) can effectively regulate blood sugar, reduce polydipsia and polyphagia symptoms in experimental animals, reduce body weight, increase glucose tolerance, improve insulin resistance, prevent complications such as cardiovascular disease, neuropathy, and nephropathy caused by diabetes, improve patients' quality of life, and reduce long-term health risks. Attached Figure Description

[0009] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of this application and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0010] Figure 1 This is a schematic diagram of the experimental procedure for the hypoglycemic effects of chrysanthemum ketone and chrysanthemum ethylsin on C57BL / 6J mice and BKS-db / db mice in the experimental examples of this application; Figure 2 The curves showing the effects of styraxone and styraxone B on fasting blood glucose in C57BL / 6J mice in the experimental examples of this application are shown. Figure 3 The experimental results of this application show the effect of styraxone and styraxone on the change of random blood glucose in C57BL / 6J mice; Figure 4 The curves showing the effects of styracil ketone and styracil ethinyl ketone on fasting blood glucose in BKS-db / db mice in the experimental examples of this application are shown. Figure 5The experimental results of this application show the effect of styracil ketone and styracil ethinyl ketone on random blood glucose levels in BKS-db / db mice. Figure 6 This is the body weight change curve of C57BL / 6J mice during the experiment in this application; Figure 7 This is a statistical graph showing the food intake of C57BL / 6J mice in the experimental cases of this application; Figure 8 This is a statistical graph showing the water consumption of C57BL / 6J mice in the experimental cases of this application; Figure 9 This is the body weight change curve of BKS-db / db mice during the experiment in this application; Figure 10 This is a statistical graph showing the food intake of BKS-db / db mice in the experimental case of this application; Figure 11 This is a statistical graph showing the water consumption of BKS-db / db mice in the experimental cases of this application; Figure 12 The effect of drug treatment on glucose tolerance in C57BL / 6J mice in this application; Figure 13 This is a statistical graph showing the area under the blood glucose concentration curve in the glucose tolerance test of C57BL / 6J mice under the drug treatment of the experimental cases in this application; Figure 14 The effect of drug administration on insulin tolerance in C57BL / 6J mice in the experimental cases of this application; Figure 15 This is a statistical graph showing the area under the blood glucose concentration curve in the insulin tolerance test of C57BL / 6J mice under the drug treatment of the experimental cases in this application; Figure 16 The effect of drug treatment on glucose tolerance in BKS-db / db mice in the experimental cases of this application; Figure 17 This is a statistical graph showing the area under the blood glucose concentration curve in the glucose tolerance test of BKS-db / db mice under the drug treatment of the experimental cases in this application; Figure 18 The effect of drug administration on insulin tolerance in BKS-db / db mice in the experimental cases of this application; Figure 19 This is a statistical graph showing the area under the blood glucose concentration curve in the insulin tolerance test of BKS-db / db mice under the drug treatment of the experimental cases in this application; Figure 20 This study applies for the effect of styraxone on the p-CREB / CREB protein level in C57BL / 6J mice; Figure 21This study applies for the effect of styracil on the p-CREB / CREB protein level in BKS-db / db mice; Figure 22 This study applies for the effect of thiamethoxam on the p-CREB / CREB protein level in C57BL / 6J mice; Figure 23 This study applies for the experimental example of the effect of fentanyl thiazolin on the p-CREB / CREB protein level in BKS-db / db mice; Figure 24 This study aims to investigate the effects of styraxone and styraxone B on the pancreas of C57BL / 6J mice. Figure 25 This study aims to investigate the effects of styraxone and styraxone B on the pancreas of BKS-db / db mice. Figure 26 For this experiment, we applied for the effect of styracil and styracil ethinylene on insulin secretion from the pancreas of C57BL / 6J mice; Figure 27 This study investigates the effects of styraxone and styraxone B on insulin secretion from the pancreas in BKS-db / db mice. Detailed Implementation

[0011] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions in the embodiments of this application will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0012] It should be noted that, unless otherwise specified, the embodiments and features described in this application can be combined with each other. The present application will now be described in detail with reference to specific embodiments.

[0013] Application of Changshan extract and its derivatives in the preparation of drugs for diabetes.

[0014] In some embodiments of this application, the above-mentioned Changshan extract is Changshan B.

[0015] Furthermore, the structural formula of dichroic acid is shown in Formula I: Formula I.

[0016] In some embodiments of this application, the derivative of the above-mentioned Changshan extract is Changshanone.

[0017] Furthermore, the structural formula of styraxone is shown in Formula II: Formula II.

[0018] The features and performance of this application will be further described in detail below with reference to the embodiments. Example

[0019] 1. Preparation of experimental mice: SPF-grade BKS-db / db mice (8 weeks old, male) and SPF-grade C57BL / 6J mice (6-7 weeks old, male) were purchased from Hangzhou Ziyuan Experimental Animal Technology Co., Ltd. and housed at Anhui Provincial Biochemical Pharmaceutical Engineering Technology Research Center. They were housed in a specific pathogen-free (SPF) environment, with sterile bedding changed every three days, and had free access to food and water.

[0020] 2. The specific details of animal grouping and drug administration are as follows: BKS-db / db mice and C57BL / 6J mice were acclimatized in the animal facility for three weeks, during which blood glucose levels were measured. The mice were then randomly divided into four groups based on blood glucose levels (saline group, metformin group, styraxone group, and styrax ethyl acetate group). Administration was performed via gavage, with the following dosages: metformin 250 mg / kg / day; styraxone 0.15 mg / kg / day; and styrax ethyl acetate 0.15 mg / kg / day. From day 18 of administration, the dosage in the styrax ethyl acetate group was adjusted from 0.15 mg / kg / day to 0.3 mg / kg / day. The metformin used was metformin hydrochloride tablets (0.5 g / tablet) manufactured by Bristol-Myers Squibb Pharmaceuticals Co., Ltd., under the brand name Glucophage. The administration of metformin, styraxone, and styrax ethyl acetate continued for 36 days. Figure 1 The diagram illustrates the experimental procedure for the hypoglycemic effect of chrysanthemum ketone or chrysanthemum ethylsin on BKS-db / db mice and C57BL / 6J mice according to embodiments of the present invention.

[0021] 3. During the drug administration period, mice were weighed every 6 days and their fasting blood glucose was measured once. Random blood glucose was measured every 7 days. After the drug administration experiment ended, mice were harvested for subsequent experiments.

[0022] 4. In this embodiment, all data were processed and statistically analyzed using Graphpad Prism 8.0 software. One-way ANOVA was used to compare sample means. p < 0.05 indicates a statistically significant difference. p<0.05 p < 0.01 p < 0.001 P < 0.0001.

[0023] 5. To investigate the effects of styracinone and styracin ethylsin on fasting and random blood glucose levels in BKS-db / db mice and C57BL / 6J mice. Figures 2-5 As shown.

[0024] in Figure 2 The following are curves showing the effects of styraxone and styraxone B on fasting blood glucose in C57BL / 6J mice according to embodiments of the present invention; Figure 3 The following are curves showing the effect of styraxone and styraxone B on random blood glucose in C57BL / 6J mice according to embodiments of the present invention; Figure 4 The following are the curves showing the effects of styracinone and styracin ethylstilbestrol on fasting blood glucose in BKS-db / db mice according to embodiments of the present invention; Figure 5 The following is a curve showing the effect of styraxone and styraxone on random blood glucose in BKS-db / db mice according to an embodiment of the present invention.

[0025] In this embodiment, fasting blood glucose was measured every 6 days during the 36-day experiment. The mice were fasted from food and water at 8 PM the previous evening, and blood was collected from the tip of the mouse's tail using a disposable lancet at 8 AM the following morning. Random blood glucose was measured every 7 days. On day 18 of drug administration, based on changes in blood glucose, the dosage of fentanyl acetate was adjusted from 0.15 mg / kg / day to 0.3 mg / kg / day.

[0026] As mentioned above Figure 4 and Figure 5 The results showed that, compared with the saline group, both the metformin group and the styraxone group effectively reduced fasting blood glucose and random blood glucose in BKS-db / db mice. The styraxone group also significantly reduced fasting blood glucose in BKS-db / db mice after adjusting the dosage from 0.15 mg / kg / day to 0.3 mg / kg / day. Figure 2 and Figure 3 The results showed that, compared with the saline group, the metformin group, styraxone group, and styraxone group had no significant difference in the effects on fasting blood glucose and random blood glucose of normal C57BL / 6J mice.

[0027] 6. Investigate the effects of styraquinone and styraquinone on body weight, diet, and water intake in C57BL / 6J and BKS-db / db mice, such as... Figures 6-11 As shown.

[0028] in, Figure 6 The figure shows the body weight change curve of C57BL / 6J mice during the experiment according to an embodiment of the present invention; Figure 7 The figure shows a statistical chart of food intake in C57BL / 6J mice according to an embodiment of the present invention. Figure 8 The figure shows the water consumption statistics of C57BL / 6J mice according to an embodiment of the present invention. Figure 9 The figure shows the body weight change curves during the BKS-db / db mouse experiment according to an embodiment of the present invention; Figure 10 The figure shows a statistical chart of food intake in BKS-db / db mice according to an embodiment of the present invention. Figure 11 The figure shows the water consumption statistics of BKS-db / db mice according to an embodiment of the present invention.

[0029] In this embodiment, the body weight of the experimental mice was measured every 6 days during the 36-day experiment. The food and water intake of the C57BL / 6J mice, BKS-db / db mice in the saline group, metformin group, styraxone group, and styraxone group was recorded every 3 days after administration, and the daily food and water intake of each mouse was calculated.

[0030] It should be noted that all mice were randomly grouped according to their fasting blood glucose levels in the first three weeks. The body weight of BKS-db / db mice on day 0 after administration was compared with the other three groups and there was no statistical difference. Figure 6 The results showed that the styraxone group and the styraxone group had no significant effect on the body weight of C57BL / 6J mice compared with the saline group. Figure 7 Metformin showed that it reduced food intake in C57BL / 6J mice with normal blood sugar levels. Figure 8 The results showed that metformin and ethionine could reduce water intake in C57BL / 6J mice. Figure 9 The results showed that, compared with the saline group, there was no significant change in body weight of BKS-db / db mice in the styraxone group and styrax ethyl group. Figure 10 The results showed that metformin and fenestrone significantly reduced food intake in BKS-db / db mice. Figure 11 The results showed that metformin, fentanyl, and fentanyl ethylsuccinate could all improve the symptoms of polydipsia in BKS-db / db mice.

[0031] 7. To investigate the effects of styraxone and styraxone on glucose tolerance and insulin resistance in C57BL / 6J mice and BKS-db / db mice.

[0032] In this embodiment, glucose tolerance and insulin tolerance tests were performed on days 28 and 30 after drug administration, respectively. The results are as follows: Figures 12-19 As shown.

[0033] The glucose tolerance test procedure was as follows: Mice were fasted and deprived of water for 12 hours. A 10% glucose solution was administered, and the mice were weighed. The oral dose was 2 g / kg. Fasting blood glucose was measured at 0 min before oral glucose administration. Blood glucose concentrations were measured at 15 min, 30 min, 60 min, 90 min, and 120 min after glucose was administered via gavage according to body weight.

[0034] The insulin tolerance test procedure was as follows: Human insulin injection solution was used. On the day of the experiment, the mice were weighed, and the injection dose was 0.5 U / kg. The mice were placed in clean cages and fasted for 4 hours. Blood glucose was measured at 0 min before intraperitoneal injection of insulin, followed by insulin injection, and blood glucose concentrations were measured at 15 min, 30 min, 60 min, 90 min, and 120 min after insulin injection.

[0035] Figure 12 The effects of drug administration treatment according to embodiments of the present invention on glucose tolerance in C57BL / 6J mice are shown. Figure 13 A statistical graph showing the area under the blood glucose concentration curve in the glucose tolerance test of C57BL / 6J mice under the drug administration treatment of the present invention is shown. Figure 14 The effects of drug administration treatment according to embodiments of the present invention on insulin tolerance in C57BL / 6J mice are shown. Figure 15 The figure shows a statistical graph of the area under the blood glucose concentration curve in the insulin tolerance test of C57BL / 6J mice under the drug administration treatment of the present invention. Figure 16 The effects of drug administration treatment according to embodiments of the present invention on glucose tolerance in BKS-db / db mice are shown. Figure 17 A statistical graph showing the area under the blood glucose concentration curve in the glucose tolerance test of BKS-db / db mice under the drug administration treatment of the present invention is shown. Figure 18 The effects of drug administration treatment according to embodiments of the present invention on insulin tolerance in BKS-db / db mice are shown. Figure 19 The figure shows a statistical graph of the area under the blood glucose concentration curve in the insulin tolerance test of BKS-db / db mice under the drug administration treatment of the present invention.

[0036] like Figures 12-15 As shown, compared with the saline group, both the styracin group and the styracin group were able to increase glucose tolerance and improve insulin resistance in BKS-db / db mice.

[0037] 8. Investigate the effects of styraxone and styraxone B on the p-CREB / CREB ratio in C57BL / 6J mice and BKS-db / db mice.

[0038] In this embodiment, after the administration was completed, mouse livers were removed, proteins were extracted, and Western blot was used to detect the effect of the administration on the p-CREB / CREB ratio of C57BL / 6J mice and BKS-db / db mice.

[0039] Figure 20 The effect of styraxone on p-CREB / CREB protein levels in C57BL / 6J mice was demonstrated. Figure 21 The effect of styraxone on the p-CREB / CREB ratio in the liver of BKS-db / db mice was demonstrated. Figure 22 The effect of diosgenin on p-CREB / CREB protein levels in C57BL / 6J mice was demonstrated. Figure 23 The effect of diosgenin on p-CREB / CREB protein levels in BKS-db / db mice was demonstrated.

[0040] like Figures 20-23 As shown, compared with the saline group, both the styracin group and the styracin group could reduce the p-CREB / CREB level in BKS-db / db mice and inhibit gluconeogenesis in mouse liver.

[0041] 9. To investigate the ameliorative effects of styraxone and styraxone on the pancreas of C57BL / 6J mice and BKS-db / db mice.

[0042] In this experimental case, after the administration was completed, the pancreas of mice was removed, and HE staining was used to observe the effect of the administration on the pancreas improvement of C57BL / 6J mice and BKS-db / db mice.

[0043] Figure 24 The pancreas of three mice is shown, demonstrating the ameliorative effect of styraxone and styraxone on the pancreas of C57BL / 6J mice. Figure 25 The image shows the pancreas of three mice, demonstrating the ameliorative effects of styraxone and styraxone on the pancreas of BKS-db / db mice.

[0044] like Figures 24-25 The results showed that styraxone and styrax ethylsin had no significant effect on the pancreas of C57BL / 6J mice, but had a certain ameliorative effect on the pancreas of BKS-db / db mice.

[0045] 10. To investigate the effects of styraxone and styraxone on insulin secretion from the pancreas of C57BL / 6J mice and BKS-db / db mice.

[0046] In this experimental case, after the administration was completed, the pancreas of mice was removed, and the effects of the administration on the pancreas of C57BL / 6J mice and BKS-db / db mice were observed using an electron microscope.

[0047] Figure 26 The pancreas of three mice is shown, demonstrating the effects of styraxone and styraxone B on insulin secretion from the pancreas of C57BL / 6J mice. Figure 27 The image shows the pancreas of three mice, demonstrating the effects of styraxone and styraxone on insulin secretion from the pancreas of BKS-db / db mice.

[0048] like Figures 26-27 As shown, both styraxone and styrax ethylsin can increase insulin secretion in the pancreas of C57BL / 6J mice and BKS-db / db mice.

[0049] The embodiments described above are some, but not all, of the embodiments of this application. The detailed description of the embodiments of this application is not intended to limit the scope of the claimed application, but merely to illustrate selected embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of this application without inventive effort are within the scope of protection of this application.

Claims

1. Application of Changshan extract and its derivatives in the preparation of drugs for diabetes.

2. The use of the Changshan extract and its derivatives according to claim 1 in the preparation of a drug for diabetes, characterized in that, The Changshan extract is Changshan B.

3. The application of the Changshan extract and its derivatives according to claim 2 in the preparation of a medicament for diabetes, characterized in that, The structural formula of the dichroic acid is shown in Formula I: Formula I.

4. The use of the Changshan extract and its derivatives according to claim 1 in the preparation of a medicament for diabetes, characterized in that, The derivative of the Changshan extract is Changshanone.

5. The use of the Changshan extract and its derivatives according to claim 4 in the preparation of a medicament for diabetes, characterized in that, The structural formula of the chrysanthemate is shown in Formula II: Formula II.

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