Application of bilobalide in preparation of medicines for preventing or treating obesity and obesity complications and preparation of anti-aging medicines or health-care products

Abstract

The invention relates to the technical field of medicines, in particular to application of bilobalide or stereoisomers thereof to preparation of medicines for preventing or treating obesity and obesity complications and preparation of anti-aging medicines or health-care products. The bilobalide disclosed by the invention can be used for effectively controlling the weight gain of mice, reducing the weight of epididymis fat of the mice, improving the holding power of the mice and increasing the muscle proportion of the mice in a high-fat feed-induced obesity model, and can be used for preparing medicines for preventing and treating diet-induced obesity, preparing anti-aging medicines or health-care products and the like.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and specifically relates to the use of bilobalide or its stereoisomers in the preparation of drugs for preventing or treating obesity and obesity complications, or in the preparation of anti-aging drugs or health products. Background Art

[0002] Obesity is a chronic disease. According to the World Health Organization's estimate, it is one of the most easily overlooked diseases that humans are currently facing, but its incidence rate is rising rapidly.

[0003] When the calories ingested by the human body are more than the calories consumed, the excess calories are stored in the body in the form of fat. When the amount exceeds the normal physiological requirement and reaches a certain value, it will evolve into obesity. In normal adult men, the weight of adipose tissue accounts for about 15% - 18% of the body weight, and in women, it accounts for about 20% - 25%. With the increase of age, the proportion of body fat increases accordingly. When the body fat increases and the weight exceeds the standard weight by 20% or the body mass index 2 (m 2 )] is greater than 24, it is called obesity. If there is no obvious cause, it is called simple obesity; if there is a clear cause, it is called secondary obesity.

[0004] Obesity brings many harms to humans. It not only affects the quality of life but also poses a great threat to health, and can cause many complications such as various cardiovascular diseases. Therefore, it is very necessary to develop some new drugs for preventing or treating obesity. Summary of the Invention

[0005] The purpose of the present invention is to provide the use of bilobalide or its stereoisomers in the preparation of drugs for preventing or treating obesity and obesity complications, or in the preparation of anti-aging drugs or health products.

[0006] The technical problems of the present invention are solved by the following technical solutions:

[0007] In the first aspect of the present invention, there is provided the use of bilobalide or its stereoisomers in the preparation of drugs for preventing or treating obesity and obesity complications.

[0008] In some embodiments of the present invention, the dosage of bilobalide is 0.1 - 400 mg / kg / day.

[0009] In some embodiments of the present invention, the dosage of bilobalide is 1 - 200 mg / kg / day.

[0010] In some embodiments of the present invention, the use of bilobalide or its stereoisomers in the preparation of drugs for preventing or treating diet-induced obesity.

[0011] In some embodiments of the present invention, the use of bilobalide or its stereoisomers in the preparation of a medicament for preventing or treating obesity caused by hypertriglyceridemia.

[0012] In some embodiments of the present invention, the use of bilobalide in the preparation of a medicament for preventing or treating complications of diet-induced obesity and obesity complications caused by hypertriglyceridemia.

[0013] In some embodiments of the present invention, the obesity complications are metabolic disorders, decreased muscle function or insulin resistance.

[0014] In some embodiments of the present invention, the metabolic disorder is diabetes.

[0015] In a second aspect of the present invention, there is provided the use of bilobalide in the preparation of an anti-aging medicament or health product.

[0016] In some embodiments of the present invention, the dosage of bilobalide is 0.1 - 400 mg / kg / day.

[0017] In some embodiments of the present invention, the dosage of bilobalide is 1 - 200 mg / kg / day.

[0018] In some embodiments of the present invention, the anti-aging medicament or health product is an anti-aging medicament or health product for increasing muscle mass.

[0019] In some embodiments of the present invention, the anti-aging medicament or health product is an anti-aging medicament or health product for increasing muscle grip strength.

[0020] The present invention includes the following beneficial effects:

[0021] Bilobalide of the present invention can effectively control the weight gain of mice and reduce the weight of epididymal fat in a diet-induced obesity model in mice, and can be used in the preparation of medicaments for preventing and treating diet-induced obesity, etc.

[0022] Bilobalide of the present invention can improve the grip strength of mice and increase the muscle mass of mice, indicating that bilobalide has a good anti-aging effect and can be used in the preparation of anti-aging medicaments or health products. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 It is the weight change curve of the diet-induced obesity model of Example 1 after 8 weeks of drug administration.

[0024] Figure 2 It is the change curve of the weight change rate of Example 1.

[0025] Figure 3 It is the Lee's index after 8 weeks of drug administration in the diet-induced obesity model of Example 1.

[0026] Figure 4 Fasting blood glucose results of the diet-induced obesity model in Example 1.

[0027] Figure 5 OGTT test results of the diet-induced obesity model in Example 1.

[0028] Figure 6 Rotarod test results of the diet-induced obesity model in Example 1.

[0029] Figure 7 Wet weight results of adipose tissue in the diet-induced obesity model in Example 1.

[0030] Figure 8 Epididymal adipocyte area in the diet-induced obesity model in Example 1.

[0031] Figure 9 Body weight change curve after 8 weeks of drug administration in the diet-induced obesity model of Example 2.

[0032] Figure 10 Body weight change rate curve in Example 2.

[0033] Figure 11 Lee's index after 8 weeks of drug administration in the diet-induced obesity model of Example 2.

[0034] Figure 12 Fasting blood glucose results of the diet-induced obesity model in Example 2.

[0035] Figure 13 OGTT test results of the diet-induced obesity model in Example 2.

[0036] Figure 14 Forelimb grip strength test results of the diet-induced obesity model in Example 2.

[0037] Figure 15 Running distance results of the diet-induced obesity model in Example 2.

[0038] Figure 16 Rotarod test results of the diet-induced obesity model in Example 2.

[0039] Figure 17 Wet weight results of adipose tissue in the diet-induced obesity model in Example 2.

[0040] Figure 18 Gastrocnemius muscle / body weight results of the diet-induced obesity model in Example 2.

[0041] Figure 19 Body weight change curve after 6 weeks of drug administration in the diet-induced hypertriglyceridemia model in Example 3.

[0042] Figure 20 It is the curve of the change rate of body weight in Example 3.

[0043] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0044] As used in the present invention, the term "prevention" refers to preventing the occurrence of a disease and / or preventing the recurrence of a disease.

[0045] The bilobalide described in the present invention can be prepared by separation and purification through the prior art.

[0046] "Stereoisomers" refer to isomers generated by different arrangements of atoms in space within a molecule, including cis-trans isomers, enantiomers, and conformational isomers. Detailed implementation manners

[0047] The specification of the present invention has described the specific implementation manners in detail. Those skilled in the art should recognize that the following implementation manners are exemplary and should not be construed as limiting the present invention. For those skilled in the art, without departing from the principle of the present invention, through several improvements and modifications to the present invention, the technical solutions obtained by these improvements and modifications also fall within the protection scope of the claims of the present invention. The beneficial effects of the present invention are specifically illustrated below through examples.

[0048] Example 1

[0049] Preventive efficacy experiment in a diet-induced obesity model of C57 mice

[0050] 1. Experimental steps

[0051] 1.1 Test animals and reagents

[0052] SPF-grade 5-week-old male C57BL / 6J mice were purchased from Chengdu Yakang Biotechnology Co., Ltd.

[0053] Bilobalide was from Chengdu Baiyu Pharmaceutical Co., Ltd.

[0054] Feed: High-fat feed 60% kcal, product number: XTHF60; maintenance feed, product number: 1010088. The feeds were all purchased from Jiangsu Xietong Pharmaceutical Biotechnology Co., Ltd.

[0055] 1.2 Establishment of a diet-induced obesity model:

[0056] Animals in the blank control group were fed maintenance feed. Nine animals were selected from the blank control group according to their body weight levels and continuously fed maintenance feed until the end of the experiment.

[0057] After the model group and each administration group maintained the transition of maintenance feed and high-fat feed at ratios of 3:7, 5:5, and 7:3 for one week, the animals were fed with high-fat feed to establish a diet-induced obesity model. After one week of diet induction, the body weight levels of the animals were measured. Compared with the animals fed with maintenance feed, the body weight of the animals increased significantly and showed statistical differences. Then, the animals were grouped. First, the animals fed with high-fat feed were sorted according to their body weight values, and 1 / 4 of the diet-induced obesity-resistant animals with lower body weight were excluded. Then, according to their body weight, they were randomly divided into 3 groups with 9 animals in each group. After grouping and administration, the animals continued to be fed with high-fat feed until the end of the experiment.

[0058] 1.3 Administration method:

[0059] After the animals were randomly grouped, administration began. The groups included: G1 blank control group (maintenance feed, Vehicle), G2 model control group (high-fat feed, Vehicle), and G4 bilobalide group (high-fat feed, 100 mg / kg). The administration volume was 5 mL / kg. The control group and the model group were gavaged with an equal volume of solvent. Vehicle: DMSO: 0.5% MC = 5:95 (v / v). Administration was performed once a day, QD, for 8 weeks.

[0060] 2. Detection indicators

[0061] 2.1 Animal body weight

[0062] Once a week, the animals were weighed at a fixed time period to monitor the change in their body weight, and the body weight change rate was calculated.

[0063] 2.2 Fasting blood glucose

[0064] Once a month, the animals were fasted overnight, and blood was collected from the tip of the tail the next day. To reduce the influence of animal stress, etc., the first drop of blood was discarded, and the subsequent blood was collected with a blood glucose test strip to read the blood glucose meter reading to monitor fasting blood glucose.

[0065] 2.3 OGTT

[0066] Before the end of the experiment, the animals were fasted overnight, administered the next day, and 30 minutes later, a glucose solution was given at 2 g / kg to establish a model. Blood was collected from the tip of the tail at 0 min, 15 min, 30 min, 60 min, and 120 min after model establishment, and the change in blood glucose was recorded, and the area under the blood glucose curve AUC was calculated.

[0067] AUC: (t 15min + t 0min ) x 0.25 / 2 + (t 30mi n + t 15min ) x 0.25 / 2 + (t 30min + t 60min ) x 0.5 / 2 + (t 120min + t 60min)x1 / 2。

[0068] 2.4 Rotarod test

[0069] Before the formal experiment, mice were subjected to rotarod training under the following conditions: Training D1: 5 r / min, 1 min; 10 r / min, 2 min; 5 - 20 r / min, 4 min, with an experimental duration of 5 min; Training D2: 5 r / min, 1 min; 10 r / min, 2 min; 5 - 30 r / min, 4 min, with an experimental duration of 5 min. During training, when the animal fell off the rotarod, it was manually placed back on the rotarod to continue training until the training duration was reached. After training, the formal experimental conditions were: 5 - 40 r / min, with an acceleration time of 4 min and a total experimental duration of 10 min. During this period, each mouse was placed individually in a rotarod compartment. Between changing mice, the rotarod was cleaned with water and alcohol to avoid odor interference with the mouse behavior. Each animal had at least a 1-hour rest interval between formal experiments. During the experiment, conversation and movement were minimized to avoid noise interference.

[0070] 2.5 Lee's index

[0071] After weighing the animals, they were anesthetized and their body lengths were measured. The body length was defined as the length from the tip of the mouse's nose to the base of the tail. When measuring the body length, the mouse was fixed to avoid arching its back, and Lee's index was calculated.

[0072] Lee's index = body weight / 3 * 1000 / body length.

[0073] 2.6 Adipose tissue

[0074] At the end of the experiment, after the animals were euthanized, bilateral epididymal fat and perirenal adipose tissue were dissected, weighed separately, and the weights of the adipose tissue were recorded. The epididymal fat was fixed, stained with H&E, and the cell area was measured and the data were statistically analyzed.

[0075] 2.7 Data statistics

[0076] The data were statistically analyzed using one-way ANOVA and post hoc tests were performed using Dunnett's. The significance labels were as follows: #P ≤ 0.05, ##P ≤ 0.01, P ≤ 0.001, model group vs. control group; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, treatment group vs. model group.

[0077] 3. Experimental results

[0078] 3.1 Effects of bilobalide on the body weight of diet-induced obesity model mice

[0079] Figure 1 This is the body weight change curve of the diet-induced obesity model in Example 1 after 8 weeks of drug administration. Figure 2It is the curve of the change rate of body weight in Example 1. Figure 3 It is the Lee's index after 8 weeks of administration of the diet-induced obesity model in Example 1.

[0080] The results showed that: before administration (D0), there was no significant difference in body weight between the model group and each administration group. Compared with the G1 blank control group, the body weights of the model group and each administration group were significantly increased, with statistical differences. At the end of the experiment, compared with the G1 control group, the body weight of the G2 model group was significantly increased. Compared with the body weight of the G2 model group, the body weights of each administration group decreased, and there was a statistical difference in the G4 bilobalide group. At the end of the experiment, compared with the initial body weight, the body weights of all groups showed an increasing trend. Compared with the G1 blank control group, the Lee's index of the G2 model group was significantly increased. Compared with the model group, the Lee's index of each administration group decreased, and there was a statistical difference in the bilobalide group. The results showed that bilobalide has a controlling effect on the body weight of high-fat diet-induced obese mice.

[0081] After the diet induction of the mice in Example 1, the initial body weight was below 25 g. At this time, the body weight was relatively small, and an obese model was not fully formed. In the subsequent experiment, the body weight of the mice in Example 1 gradually increased, forming a stable obese model. The model in Example 1 was a preventive administration model, indicating that bilobalide can prevent obesity.

[0082] 3.2 Effect of bilobalide on fasting blood glucose in diet-induced obese model mice

[0083] Figure 4 It is the result of fasting blood glucose of the diet-induced obesity model in Example 1.

[0084] The results showed that: after administration to D42, compared with the G1 blank control group, the fasting blood glucose of the G2 model control group was significantly increased, with statistical differences; compared with the G2 model control group, the fasting blood glucose of the G3 statin group and the G4 bilobalide group decreased, and there was a statistical difference in the bilobalide group, indicating that bilobalide has a controlling effect on the fasting blood glucose of obese mice.

[0085] 3.3 Effect of bilobalide on oral glucose tolerance in diet-induced obese model mice

[0086] Figure 5 It is the result of the OGTT experiment of the diet-induced obesity model in Example 1. The results showed that: compared with the G1 blank control group, after glucose modeling in the G2 model control group, the blood glucose increased rapidly and decreased slowly, and the area under the blood glucose curve AUC had statistical differences, indicating that the glucose tolerance of the model group was impaired; compared with the G2 model control group, the G4 bilobalide group improved the regulatory ability of the mouse body to blood glucose concentration, and the AUC decreased, with statistical differences.

[0087] 3.4 Effect of bilobalide on the staying time on the rod in diet-induced obese model mice

[0088] Figure 6 Rotarod test results of the diet-induced obesity model in Example 1.

[0089] The results showed that compared with the G1 blank control group, the rod duration of the G2 model control group decreased, and there was a statistically significant difference, indicating that the endurance or motor coordination performance of the model group was impaired; compared with the G2 model control group, the rod duration of the mice in each drug administration group showed an upward trend, indicating that bilobalide had an improvement effect on the impairment of motor coordination or endurance caused by obesity.

[0090] 3.5 Effect of bilobalide on adipose tissue of mice in the diet-induced obesity model

[0091] Figure 7 Results of the wet weight of adipose tissue in the diet-induced obesity model of Example 1 Figure 8 Epididymal fat cell area of the diet-induced obesity model in Example 1

[0092] The results showed that compared with the G1 blank control group, the wet weight of adipose tissue in the G2 model control group was significantly increased, and the fat cell area was significantly enlarged; compared with the G2 model control group, the wet weight of adipose tissue in the G4 bilobalide group decreased, and the fat cell area decreased. The data showed that bilobalide had a controlling effect on the weight and cell area of adipose tissue in animals of the diet-induced obesity model.

[0093] In summary, it shows that: in the obesity model induced by high-fat diet, bilobalide of the present invention can effectively control the body weight gain of mice, control the blood glucose of mice, improve the glucose tolerance of mice, increase the rod duration of mice, reduce the fat weight and cell area of mice, and can be used for preparing drugs for preventing and treating complications such as diet-induced obesity, insulin resistance, type 2 diabetes, and decline in motor function.

[0094] Example 2

[0095] Therapeutic efficacy experiment in the C57 mouse diet-induced obesity model

[0096] 1. Experimental steps

[0097] 1.1 Test animals and reagents

[0098] SPF-grade 5-week-old male C57BL / 6J mice were purchased from Chengdu Yakang Biotechnology Co., Ltd.

[0099] Bilobalide was from Chengdu Baiyu Pharmaceutical Co., Ltd.

[0100] Feed: High-fat feed 60% kcal, product number: XTHF60; maintenance feed, product number: 1010088, and the feeds were all purchased from Jiangsu Xietong Pharmaceutical Biotechnology Co., Ltd.

[0101] 1.2 Establishment of diet-induced obesity model:

[0102] Animals in the blank control group were fed maintenance feed. Eight animals were selected from the blank control group according to their body weight levels and continuously fed maintenance feed until the end of the experiment.

[0103] After one week of transition with the maintenance feed and high-fat feed in the ratio of 3:7, 5:5, and 7:3 for the model group and each drug administration group respectively, the animals were fed high-fat feed to establish a diet-induced obesity model. After one week of diet induction, the body weight levels of the animals were measured. Compared with the animals fed maintenance feed, the body weight of the animals increased significantly and showed statistical differences. Then, the animals were grouped. First, the animals fed high-fat feed were sorted according to their body weight values, and 1 / 4 of the diet-induced obesity-resistant animals with lower body weight were excluded. Then, they were randomly divided into 3 groups of 8 animals each according to the S-shaped grouping method based on body weight. After grouping and drug administration, they continued to be fed high-fat feed until the end of the experiment.

[0104] 1.3 Drug administration method:

[0105] After random grouping, the animals started drug administration. The groups included: G1 blank control group (maintenance feed, Vehicle), G2 model control group (high-fat feed, Vehicle), G3 bilobalide group (high-fat feed, 100 mg / kg), and G4 positive control orlistat group (high-fat feed, 20 mg / kg). The drug administration volume was 5 mL / kg. The control group and the model group were gavaged with an equal volume of solvent. Vehicle: DMSO: 0.5% MC = 5:95 (v / v). The drug was administered by gavage once a day, QD, for 8 weeks.

[0106] 2. Detection indicators

[0107] 2.1 Animal body weight

[0108] Once a week, the animals were weighed at a fixed time period to monitor the change in body weight and calculate the body weight change rate.

[0109] 2.2 Fasting blood glucose

[0110] Once a month, the animals were fasted overnight, and blood was collected from the tip of the tail on the second day. To reduce the influence of animal stress, etc., the first drop of blood was discarded, and the subsequent blood was collected with a blood glucose test strip, and the reading of the blood glucose meter was recorded to monitor the fasting blood glucose.

[0111] 2.3 OGTT

[0112] Before the end of the experiment, the animals were fasted overnight, and drugs were administered on the second day. After 30 minutes, a glucose solution was given at 2 g / kg for model establishment, and blood was collected from the tip of the tail at 0 min, 15 min, 30 min, 60 min, and 120 min after model establishment. The change in blood glucose was recorded, and the area under the blood glucose curve AUC was calculated.

[0113] AUC: (t 15min + t 0min ) x 0.25 / 2 + (t 30mi n + t 15min ) x 0.25 / 2 + (t 30min + t 60min ) x 0.5 / 2 + (t 120min + t 60min ) x 1 / 2。

[0114] 2.4 Forelimb Grip Strength

[0115] Before the end of the experiment, grip strength tests were conducted. The animals were allowed to adapt to the testing instrument and restraint method in advance. Before the test, the animal weights were measured. The animal's tail was grasped, and it was allowed to grasp the digital dynamometer. At the same time, the animal was gently pulled parallel to the test rod by the tail. When the maximum force was applied to the experimental mice, the reading on the grip strength meter was recorded. Multiple repeated measurements were taken to ensure the results, and force / body weight (g / g) was calculated.

[0116] 2.5 Treadmill Test

[0117] Before the formal experiment, the mice were subjected to running training under the following conditions: D1 training: free movement on a stationary track for 10 min; 5 m / min for 5 min; 8 m / min for 5 min; D2 training: 5 m / min for 4 min; 8 m / min for 3 min; 12 m / min for 3 min; D3 training: 5 m / min for 4 min; 10 m / min for 3 min; 15 m / min for 3 min. During the training, the number of electric shocks did not exceed 5 times, and the duration did not exceed 1 s. When the animal left the track, it was forced back to the track by the experimenter to continue training. After the training, the formal experimental conditions were as follows: 10 m / min, with an acceleration duration of 3 min and a maintenance duration of 5 min, then directly 18 m / min, with an acceleration duration of 15 min and a maintenance duration of 900 s, electric shock for 10 s, 50 times, 0.4 mA. During this period, each mouse was placed separately in a single track. After each mouse, the track was cleaned with water and alcohol to avoid odor interference with the mouse's behavior. There was at least a 1-h rest interval between each formal experiment for the animals. During the experiment, conversations and movements were minimized to avoid noise interference.

[0118] 2.6 Rotarod Test

[0119] Before the formal experiment, the mice were subjected to rotarod training, and the training conditions were as follows: Training D1: 5 r / min, 1 min; 10 r / min, 1 min; 5 - 20 r / min, 3 min, with an experimental duration of 5 min; Training D2: 5 r / min, 1 min; 10 r / min, 1 min; 5 - 30 r / min, 3 min, with an experimental duration of 5 min. During the training, when the animal fell off the rotarod, it was manually placed back on the rotarod to continue training until the training duration was reached. After the training, the formal experimental conditions were: 5 - 40 r / min, with an acceleration time of 5 min and a total experimental duration of 10 min. During this period, each mouse was placed separately in a rotarod compartment. When changing mice, the rotarod needed to be cleaned with water and alcohol to avoid odor interference with the mouse behavior. The animals had at least a 1 - hour rest interval between each formal experiment. During the experiment, conversations and movements were minimized to avoid noise interference.

[0120] 2.7 Lee's index

[0121] At the end of the experiment, the animals were weighed and their body lengths were measured. The body length was defined as the length from the tip of the mouse's nose to the base of the tail. When measuring the body length, the mouse was fixed to avoid arching its back, and the Lee's index was calculated.

[0122] Lee's index = body weight / 3 * 1000 / body length.

[0123] 2.8 Tissue weighing

[0124] At the end of the experiment, after the animals were euthanized, the bilateral epididymal fat, perirenal fat, and gastrocnemius muscle tissues were dissected and weighed separately. The weight of the adipose tissue was recorded, and the gastrocnemius muscle / body weight (mg / g) ratio was calculated.

[0125] 2.9 Data statistics

[0126] The data were statistically analyzed by one - way ANOVA and post - hoc test was performed using Dunnett's. The significance markers are as follows: #P ≤ 0.05, ##P ≤ 0.01, P ≤ 0.001, model group vs control group; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, treatment group vs model group.

[0127] 3. Experimental results

[0128] 3.1 Effect of bilobalide on the body weight of diet - induced obese model mice

[0129] Figure 9 It is the body weight change curve of the diet - induced obese model in Example 2 after 8 - week administration. Figure 10 It is the change rate curve of the body weight in Example 2. Figure 11 It is the Lee's index of the diet - induced obese model in Example 2 after 8 - week administration.

[0130] The results showed that: before administration (D0), there was no significant difference in body weight among the model group and each administration group. Compared with the G1 blank control group, the body weights of the model group and each administration group were significantly increased, showing statistical differences. At the end of the experiment, compared with the G1 control group, the body weight of the G2 model group was significantly increased. Compared with the body weight of the G2 model group, the body weights of each administration group decreased, and there was a statistical difference in the G3 bilobalide group. At the end of the experiment, compared with the initial body weight, the body weights of all groups showed an increasing trend. Compared with the G1 blank control group, the Lee's index of the G2 model group was significantly increased. Compared with the model group, the Lee's index of each administration group decreased, and there was a statistical difference in the bilobalide group. The results showed that bilobalide had a controlling effect on the body weight of high-fat diet-induced obese mice.

[0131] After diet induction in the mice of Example 2, the initial body weight at the start of the experiment was above 25 g, and the body weight was relatively large at this time, forming a stable obesity model. The model of Example 2 was a treatment administration model, indicating that bilobalide could treat obesity.

[0132] 3.2 Effect of bilobalide on fasting blood glucose in diet-induced obese model mice

[0133] Figure 12 This is the result of fasting blood glucose in the diet-induced obese model of Example 2.

[0134] The results showed that: until D43 of administration, compared with the G1 blank control group, the fasting blood glucose of the G2 model control group increased significantly, showing statistical differences; compared with the G2 model control group, the fasting blood glucose of the G3 bilobalide group and the G4 orlistat group decreased, showing statistical differences, indicating that bilobalide had a controlling effect on the fasting blood glucose of obese mice.

[0135] 3.3 Effect of bilobalide on oral glucose tolerance in diet-induced obese model mice

[0136] Figure 13 This is the result of the OGTT experiment in the diet-induced obese model of Example 2.

[0137] The results showed that: compared with the G1 blank control group, after glucose modeling in the G2 model control group, the blood glucose increased rapidly and decreased slowly, and the area under the blood glucose curve AUC showed statistical differences, indicating that the glucose tolerance of the model group was impaired; compared with the G2 model control group, the G3 bilobalide group improved the regulatory ability of the mouse body to blood glucose concentration, and the AUC decreased, showing statistical differences. Under this administration condition, there was no obvious improvement in glucose tolerance in the G4 orlistat group.

[0138] 3.4 Effect of bilobalide on the forelimb grip strength of diet-induced obese model mice

[0139] Figure 14 This is the result of the forelimb grip strength test in the diet-induced obese model of Example 2.

[0140] The results showed that compared with the G1 blank control group, the grip strength and the ratio of grip strength to body weight in the G2 model control group both decreased, and there were statistical differences, indicating that the muscle strength in the model group was impaired; compared with the G2 model control group, the forelimb grip strength and the ratio of grip strength to body weight of mice in each drug administration group increased, and there were statistical differences, indicating that bilobalide improved the muscle strength impairment caused by obesity and increased the muscle grip strength.

[0141] 3.5 Effect of bilobalide on the running distance of mice in a diet-induced obesity model

[0142] Figure 15 The running distance results of the diet-induced obesity model in Example 2 are shown.

[0143] The results showed that compared with the G1 blank control group, the running distance of the G2 model control group decreased; compared with the G2 model control group, the running distances of the G3 bilobalide group and the G4 orlistat group increased. The data indicated that the bilobalide group had a certain improvement effect on the endurance of animals in the diet-induced obesity model.

[0144] 3.6 Effect of bilobalide on the time on the rod of mice in a diet-induced obesity model

[0145] Figure 16 The results of the rotarod test of the diet-induced obesity model in Example 2 are shown.

[0146] The results showed that compared with the G1 blank control group, the time on the rod of the G2 model control group decreased, indicating that the endurance or motor coordination performance of the model group was impaired; compared with the G2 model control group, the time on the rod of the G3 bilobalide group and the G4 orlistat group both showed an upward trend, indicating that bilobalide had an improvement effect on the impairment of motor coordination or endurance caused by obesity.

[0147] 3.7 Effect of bilobalide on the tissue weight of mice in a diet-induced obesity model

[0148] Figure 17 The wet weight results of adipose tissue of the diet-induced obesity model in Example 2 are shown. Figure 18 The results of gastrocnemius muscle / body weight of the diet-induced obesity model in Example 2 are shown.

[0149] The results showed that compared with the G1 blank control group, the wet weight of adipose tissue in the G2 model control group increased significantly, and the gastrocnemius muscle / body weight decreased significantly; compared with the G2 model control group, the wet weight of adipose tissue in the G3 bilobalide group and the G4 orlistat group decreased, and the gastrocnemius muscle / body weight increased, and there were statistical differences in the bilobalide group. The data indicated that bilobalide had a controlling effect on the adipose tissue weight of animals in the diet-induced obesity model, and could improve the proportion of muscle tissue and increase the muscle proportion.

[0150] In summary, the above description shows that in the obesity model induced by high-fat diet, bilobalide can effectively control the weight gain of mice, control blood glucose, improve glucose tolerance, and can be used for preparing drugs for preventing and treating obesity or obesity complications such as diet-induced obesity, insulin resistance, type 2 diabetes, etc. In the obesity model induced by high-fat diet, bilobalide can increase the grasping force and running distance of mice, improve the proportion of muscle tissue, reduce the fat weight and muscle ratio of mice, and can be used for preventing and treating the decline of muscle function, etc., so as to achieve the anti-aging effect.

[0151] Example 3

[0152] Pharmacodynamic experiment in the hypertriglyceridemia model of golden hamsters

[0153] 1. Experimental procedures

[0154] 1.1 Test animals and reagents

[0155] SPF-grade 7-week-old male golden hamsters (strain name: LVG Hamster, code: 501) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0156] Bilobalide was from Chengdu Baiyu Pharmaceutical Co., Ltd.

[0157] Feed: High-fructose feed 60% kcal%, product number: XT-704; maintenance feed, product number: 1010088. The feeds were all purchased from Jiangsu Xietong Pharmaceutical Bioengineering Co., Ltd.

[0158] 1.2 Establishment of the hypertriglyceridemia model by diet induction:

[0159] Animals in the blank control group were fed maintenance feed. 8 animals were selected for the blank control group according to the serum triglyceride level and continued to be fed maintenance feed until the end of the experiment.

[0160] Animals in the model group and each drug administration group were fed high-fructose feed. After the maintenance feed and high-fructose feed were transitioned according to 3:7, 5:5, and 7:3, the animals were fed high-fructose feed to establish the hypertriglyceridemia model. After one to two weeks of diet induction, the serum triglyceride level was detected. Compared with the animals fed maintenance feed, the serum triglyceride was significantly increased and there was a statistical difference. Then, the animals were grouped, and animals with serum triglyceride ≥ 2.3 mmol / L were selected. According to the serum triglyceride level, they were randomly divided into 3 groups with 8 animals in each group. After grouping and drug administration, they continued to be fed high-fructose feed until the end of the experiment.

[0161] 1.3 Drug administration method:

[0162] After randomly grouping the animals, drug administration was started. The groups included: G1 blank control group (maintenance diet, Vehicle), G2 model control group (high-fructose diet, Vehicle), G3 high-dose bilobalide group (high-fructose diet, 75 mg / kg), and G4 low-dose bilobalide group (high-fructose diet, 25 mg / kg). The administration volume was 5 mL / kg. The control group and the model group were gavaged with an equal volume of solvent. Vehicle: DMSO: 0.5% MC = 5:95 (v / v). The drug was administered by gavage once a day, QD, for 6 weeks.

[0163] 2. Detection indicators

[0164] 2.1 Animal body weight

[0165] Twice a week, the body weight changes of the animals were monitored, and the body weight change rate was calculated.

[0166] 3. Experimental results

[0167] 3.1 Effect of bilobalide on the body weight of diet-induced hypertriglyceridemia model golden hamsters

[0168] Figure 19 It is the body weight change curve after 6 weeks of drug administration for the diet-induced hypertriglyceridemia model in Example 3. Figure 20 It is the change curve of the body weight change rate in Example 3.

[0169] The results showed that: before drug administration (D0), there was no significant difference in body weight among the groups. By the last observation before the end of drug administration (D39), compared with the G2 model control group, the body weights of the G3 and G4 high- and low-dose bilobalide groups decreased after drug administration, showing a dose-dependent manner. At the end of the experiment, compared with the initial body weight, the body weights of all groups showed an increasing trend. Bilobalide has a controlling effect on the body weight of golden hamsters induced by a high-fructose diet.

[0170] In summary, it shows that: in the high-fructose-induced hypertriglyceridemia model, bilobalide can effectively control body weight gain and can be used to prepare drugs for preventing or treating obesity caused by hypertriglyceridemia, etc.

[0171] The specification of the present invention has described the specific implementation manners in detail. Those skilled in the art should recognize that the above implementation manners are exemplary and should not be construed as limitations on the present invention. For those skilled in the art, without departing from the principle of the present invention, by making several improvements and modifications to the present invention, the technical solutions obtained by these improvements and modifications also fall within the protection scope of the claims of the present invention.

Claims

1. Use of bilobalide or its stereoisomers in the preparation of a medicament for preventing or treating obesity.

2. The use according to claim 1, characterized in that: The dosage of bilobalide is 0.1 - 400 mg / kg / day; preferably, the dosage of bilobalide is 1 - 200 mg / kg / day.

3. The use according to claim 1, characterized in that: Use of bilobalide or its stereoisomers in the preparation of a medicament for preventing or treating diet-induced obesity.

4. The use according to claim 1, characterized in that: Use of bilobalide or its stereoisomers in the preparation of a medicament for preventing or treating obesity caused by hypertriglyceridemia.

5. The use according to claim 3 or 4, characterized in that: Use of bilobalide in the preparation of a medicament for preventing or treating complications of diet-induced obesity and complications of obesity caused by hypertriglyceridemia.

6. The use according to claim 5, characterized in that: The complications of obesity are metabolic disorders, muscle function decline or insulin resistance.

7. Use of bilobalide in the preparation of an anti-aging medicament or health product.

8. The use according to claim 7, characterized in that: The dosage of bilobalide is 0.1 - 400 mg / kg / day; preferably, the dosage of bilobalide is 1 - 200 mg / kg / day.

9. The use according to claim 7 or 8, characterized in that: The anti-aging medicament or health product is an anti-aging medicament or health product for increasing muscle proportion.

10. Use according to claim 7 or 8, characterized in that: The anti-aging medicament or health product is an anti-aging medicament or health product for increasing muscle grip strength.

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