阅读说明：本技术 正丁基丙二酸二乙酯在制备防治肥胖药物中应用 (Application of diethyl n-butylmalonate in preparation of obesity prevention and treatment medicines ) 是由 于英华 潘伟 赵进秀 蒋鹏飞 徐大祥 杨晓莹 张鹏 胡敏敏 胡涛 李梦迪 于 2021-09-24 设计创作，主要内容包括：本发明提供了正丁基丙二酸二乙酯在制备防治肥胖药物中应用,属于生物医学技术领域。正丁基丙二酸二乙酯能够降低肥胖小鼠体重、肝脏重量,防止肝组织脂质累积,增强肥胖小鼠机体脂肪酸氧化、增加能量消耗,从而发挥减肥效果；而且能够改善葡萄糖耐受和胰岛素抵抗,具有纠正肥胖相关代谢紊乱的作用。(The invention provides application of diethyl n-butylmalonate in preparation of a medicament for preventing and treating obesity, and belongs to the technical field of biomedicine. The diethyl n-butylmalonate can reduce the weight and the liver weight of the obese mouse, prevent the accumulation of liver tissue lipid, enhance the oxidation of fatty acid in the organism of the obese mouse and increase the energy consumption, thereby exerting the weight-losing effect; and can improve glucose tolerance and insulin resistance, and has effect in correcting obesity-related metabolic disorder.)

1. Application of diethyl n-butylmalonate in preparing medicine for preventing and treating obesity is provided.

2. The use of diethyl n-butylmalonate in the preparation of a medicament for preventing and treating obesity according to claim 1, wherein: the obesity preventing and treating medicine comprises a weight-losing medicine.

3. The use of diethyl n-butylmalonate in the preparation of a medicament for preventing and treating obesity according to claim 1, wherein: the medicine for preventing and treating obesity comprises a medicine for preventing and treating liver tissue lipid accumulation.

4. The use of diethyl n-butylmalonate in the preparation of a medicament for preventing and treating obesity according to claim 1, wherein: the obesity preventing and treating medicine comprises a medicine for improving glucose tolerance.

5. The use of diethyl n-butylmalonate in the preparation of a medicament for preventing and treating obesity according to claim 1, wherein: the obesity preventing and treating medicine comprises a medicine for improving insulin resistance.

6. The use of diethyl n-butylmalonate in the preparation of a medicament for preventing and treating obesity according to claim 1, wherein: the medicine for preventing and treating obesity comprises medicines for enhancing body fatty acid oxidation and increasing energy consumption.

7. Use of diethyl n-butylmalonate according to claims 1-6 in the preparation of a medicament for the prevention and treatment of obesity, characterized in that: the medicament for preventing and treating obesity takes diethyl n-butylmalonate as an active ingredient and is supplemented with pharmaceutically acceptable auxiliary materials or carriers.

Technical Field

The invention belongs to the technical field of biomedicine, and particularly relates to application of diethyl n-butylmalonate in preparation of a medicament for preventing and treating obesity.

Background

Obesity is an epidemic, systemic metabolic disease that seriously harms human health. Obesity is a metabolic syndrome in which energy metabolism is abnormal, fat accumulation in a body is excessive, and distribution is abnormal due to various factors such as heredity, lifestyle, and immune state. It is closely related to insulin resistance, diabetes, hyperlipidemia, hypertension, fatty liver, cardiovascular diseases and cognitive disorder. Most of the prior weight-reducing methods have the defects of large side effect, high recurrence rate and the like, and particularly, a large number of weight-reducing medicines are released from the market due to the safety problem. In view of the worldwide increasing incidence of obesity and metabolic diseases caused by obesity, a safe and effective novel prevention and treatment strategy is sought, and the method has important significance for reducing the incidence of obesity, intervening in obesity-related health problems and guaranteeing the life health of people.

Succinate dehydrogenase is a metabolic enzyme in the cellular mitochondrial tricarboxylic acid cycle. The research finds that the expression up-regulation or activity increase of the enzyme can promote the progress of the Parkinson disease, so the enzyme is regarded as a potential drug target for intervening neurodegenerative diseases. Recent studies have also shown that succinate dehydrogenase is also involved in the regulation of macrophage-induced inflammatory responses.

Diethyl n-butylmalonate is a known competitive inhibitor of succinate dehydrogenase. Recent evidence at in vitro levels suggests that the inhibitors may down-regulate lipopolysaccharide-induced macrophage inflammation and thus have anti-inflammatory effects. However, whether the traditional Chinese medicine can prevent and treat obesity has not been reported.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention aims to provide the application of diethyl n-butylmalonate in preparing the medicament for preventing and treating obesity, which can prevent and treat obesity.

The invention is realized by the following technical scheme: application of diethyl n-butylmalonate in preparing medicine for preventing and treating obesity is provided.

The preferable scheme of the application of the diethyl n-butylmalonate in preparing the medicament for preventing and treating the obesity comprises the following steps: the obesity preventing and treating medicine comprises a weight-losing medicine.

The preferable scheme of the application of the diethyl n-butylmalonate in preparing the medicament for preventing and treating the obesity comprises the following steps: the medicine for preventing and treating obesity comprises a medicine for preventing and treating liver tissue lipid accumulation.

The preferable scheme of the application of the diethyl n-butylmalonate in preparing the medicament for preventing and treating the obesity comprises the following steps: the obesity preventing and treating medicine comprises a medicine for improving glucose tolerance.

The preferable scheme of the application of the diethyl n-butylmalonate in preparing the medicament for preventing and treating the obesity comprises the following steps: the obesity preventing and treating medicine comprises a medicine for improving insulin resistance.

The preferable scheme of the application of the diethyl n-butylmalonate in preparing the medicament for preventing and treating the obesity comprises the following steps: the medicine for preventing and treating obesity comprises medicines for enhancing body fatty acid oxidation and increasing energy consumption.

The preferable scheme of the application of the diethyl n-butylmalonate in preparing the medicament for preventing and treating the obesity comprises the following steps: the medicament for preventing and treating obesity takes diethyl n-butylmalonate as an active ingredient and is supplemented with pharmaceutically acceptable auxiliary materials or carriers.

The invention has the beneficial technical effects that: the invention provides application of diethyl n-butylmalonate in preparation of a medicament for preventing and treating obesity. In addition, diethyl n-butylmalonate can improve glucose tolerance and insulin resistance, and has the effect of correcting obesity-related metabolic disorders.

The results of the embodiments of the present invention show that: diethyl n-butylmalonate can prevent lipid accumulation in liver tissue, increase fatty acid oxidation and energy consumption of organism, prevent and treat obesity, improve glucose tolerance and insulin resistance, and improve obesity-related metabolic disorder.

Drawings

FIG. 1 shows the effect of diethylbutylmalonate on the body weight and cumulative body weight (curve) of mice;

FIG. 2 is a graph showing the effect of diethylbutylmalonate on body weight of mice at the time of killing;

FIG. 3 is a graph showing the effect of diethylbutylmalonate on mouse liver weight;

FIG. 4 is a photograph of pathological sections (x400) showing the effect of diethylbutylmalonate on mouse liver histomorphology;

FIG. 5 is a graph of the effect of diethylbutylmalonate on mouse oral glucose tolerance;

FIG. 6 shows the effect of diethylbutylmalonate on fasting insulin levels and HOMA-IR index in mice;

FIG. 7 is a graph of the effect of diethyl n-butylmalonate on mouse energy expenditure;

FIG. 8 is a graph of the effect of diethyl n-butylmalonate on mouse fatty acid oxidation;

FIG. 9 is a graph of the effect of diethyl n-butylmalonate on energy intake in mice.

Detailed Description

The invention is further illustrated by the following examples and figures.

The invention provides application of diethyl n-butylmalonate in preparing a medicament for preventing and treating obesity.

In the invention, the diethyl n-butylmalonate can reduce the weight of the mouse and the weight of the liver, reduce the accumulation of liver tissues, promote the oxidation of fatty acid and the energy consumption of the mouse, and further can prevent and treat obesity. In the invention, the diethyl n-butylmalonate can prevent the accumulation of liver tissue lipid, can improve glucose tolerance and insulin resistance, and has the effect of correcting obesity-related metabolic disorders.

The diethyl n-butylmalonate is prepared into the anti-obesity drug, wherein the diethyl n-butylmalonate in the anti-obesity drug is administrated according to 25 mg/kg of body weight, the administration method is preferably intraperitoneal injection, and the number of injections per week is preferably 2; the time of injection is not particularly limited.

In the invention, the medicament for preventing and treating obesity takes diethyl n-butylmalonate as an active ingredient, and can be supplemented with pharmaceutically acceptable auxiliary materials or carriers.

In the present invention, the obesity prevention and treatment drug further includes a drug for preventing lipid accumulation in liver tissue. In the invention, the diethylbutylmalonate in the anti-obesity drug is administered at 25 mg/kg body weight; the administration method of the medicine for preventing and treating obesity is preferably intraperitoneal injection, and the frequency of injection in each week is preferably 2 times; the time of injection is not particularly limited. In the invention, the medicament for preventing and treating obesity takes diethyl butylmalonate as an active ingredient, and can be supplemented with pharmaceutically acceptable auxiliary materials or carriers.

In the present invention, the obesity preventive drug further includes a drug for improving glucose tolerance. In the invention, the diethylbutylmalonate in the anti-obesity drug is administered at 25 mg/kg body weight; the administration method of the medicine for preventing and treating obesity is preferably intraperitoneal injection, and the number of injections per week is preferably 2. The present invention is not particularly limited with respect to the time of injection. In the invention, the medicament for preventing and treating obesity takes diethyl butylmalonate as an active ingredient, and can be supplemented with pharmaceutically acceptable auxiliary materials or carriers.

In the present invention, the obesity prevention and treatment drugs further include drugs that improve insulin resistance. In the invention, the diethylbutylmalonate in the anti-obesity drug is administered at 25 mg/kg body weight; the administration method of the medicine for preventing and treating obesity is preferably intraperitoneal injection, and the number of injections per week is preferably 2. The present invention is not particularly limited with respect to the time of injection. In the invention, the medicament for preventing and treating obesity takes diethyl butylmalonate as an active ingredient, and can be supplemented with pharmaceutically acceptable auxiliary materials or carriers.

In the present invention, the obesity prevention and treatment drugs further include drugs that enhance fatty acid oxidation and energy expenditure. In the present invention, the drug is administered with 25 mg/kg body weight of diethyl n-butylmalonate; the administration method of the medicine for preventing and treating obesity is preferably intraperitoneal injection, and the number of injections per week is preferably 2. The present invention is not particularly limited with respect to the time of injection. In the invention, the medicament for preventing and treating obesity takes diethyl butylmalonate as an active ingredient, and can be supplemented with pharmaceutically acceptable auxiliary materials or carriers.

The application of diethyl n-butylmalonate in the preparation of the medicament for preventing and treating obesity provided by the invention is described in detail by the following examples, but the application of diethyl n-butylmalonate in the preparation of the medicament for preventing and treating obesity is not to be construed as limiting the scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Pharmacodynamic study on improvement of obesity and related metabolic diseases by diethyl n-butylmalonate

First, experiment method

1. Design of animal experiments

The 7-week-old C57BL/6J mice were purchased from Beijing Wittingle laboratory animal technology, Inc., and were housed in the laboratory animal center of Xuzhou medical university under 12 h light-dark alternation at a temperature of 23 + -3 deg.C and a relative humidity of 50%. After 1 week of adaptive feeding, mice were randomized into 4 groups (n-10) designed to be:

LC group: conventional feed feeding, i.e. 100. mu.l of sterile PBS solution per mouse is injected into abdominal cavity 2 times per week;

LC + DM group: feeding with conventional feed, injecting 100 μ l PBS solution containing diethyl n-butylmalonate (DM, Cat. STBB9585V, Sigma-Aldrich) into abdominal cavity of each mouse, and administering 25 mg/kg per mouse 2 times per week;

HF group: feeding with high fat feed, injecting 100 μ l sterile PBS solution into abdominal cavity of mouse, and 2 times per week;

HF + DM group: each mouse was injected intraperitoneally with 100. mu.l of sterile PBS containing DM 2 times per week, 25 mg/kg per mouse.

Weigh once every 7 days, mice were free to eat and drink water during the experiment; the method comprises the following steps of fasting a mouse for 12 hours before the experiment is finished, weighing the weight, conducting cervical dislocation and sacrifice on the mouse after blood is taken from an orbit, rapidly picking out liver, brown fat, subcutaneous fat and brown fat, taking a small part of fat and liver tissues to perform fixed preservation in 4% paraformaldehyde, placing the rest tissues in a refrigerator at minus 80 ℃, centrifuging a blood sample at 8000 rpm for 5 min, and taking serum to preserve.

2. Small animal energy metabolism monitoring

At week 13 of the experiment, the respiratory quotient, heat production, oxygen consumption, carbon dioxide (CO) were monitored over 24 h using a small animal energy metabolizer (Oxymax CLAMS system)₂) Amount of produced, oxygen (O)₂) The amount produced. The following indices are calculated according to the formula:

carbohydrate oxidation level = [ (4.585 CO =)₂Production amount) - (3.226 x O₂Production amount)]*4；

Fat oxidation level = [ (1.695 x O)₂Production amount) - (1.701 × CO₂Production amount)]*9。

3. Oral Glucose Tolerance Test (OGTT)

At week 11 of the experiment, a glucose tolerance test was performed. Before detection, mice are fasted for 16 h, and glucose is injected into the abdominal cavity, wherein the injection amount is 2g/kg body weight. Whole blood was obtained by a rat tail blood sampling method, and blood glucose concentrations (mmol/L) were measured with a blood glucose meter at 0, 15 min, 30 min, 45 min, 1 h and 2 h after injection, respectively.

4. HOMA-IR assay

At week 12 of the experiment, steady-state model insulin resistance index (HOMA-IR) measurements were performed. After fasting for 6 hours, whole blood was collected by a rat tail blood collection method, and fasting blood glucose (mmol/L) and fasting insulin (mU/L) were measured by an ELISA method. HOMA-IR = blood glucose concentration (mmol/L) x insulin concentration (mU/L)/22.5 was calculated.

5. Pathological analysis

Oil red O staining analysis: liver tissues of each group of animals were taken, fixed with 4% paraformaldehyde, paraffin-embedded, sectioned, dehydrated, fixed, stained with oil red O, and histopathological analysis was performed under a microscope.

6. Statistical analysis

Experimental data were analyzed for variance and significance using SPSS 22.0 statistical software. Normally distributed metrology data is expressed as mean ± standard deviation. Through the test of normality and homogeneity of variance, the difference comparison among a plurality of groups of metering data with normality and homogeneity of variance adopts one-way analysis of variance (ANOVA), and the two-by-two comparison among each group is tested by a Turkey's method. Inspection level α = 0.05.

Second, experimental results

1. Diethyl n-butylmalonate can reduce the weight of a mouse and increase the accumulated weight

As can be seen from fig. 1, the experimental mice did not develop abnormal clinical symptoms during the course of the 16-week experiment, and after the induction of high fat diet for 15 weeks, the high fat group (HF) mice developed significant obesity compared to the normal control group (LC); the weight and the accumulated weight gain of obese mice (HF + DM) affected by diethyl n-butylmalonate were significantly lower than those of the HF group, and diethyl n-butylmalonate had no significant effect on the weight and the accumulated weight proliferation of normal mice (FIG. 1). Experimental results show that the diethyl n-butylmalonate can effectively slow down the weight increase of mice caused by high-fat diet.

2. The diethyl n-butylmalonate can reduce the weight of mice and the weight of livers

Mice were killed by dissection at week 16. Weighing before killing revealed that diethyl n-butylmalonate significantly reduced the body weight of obese mice (fig. 2). When the livers were taken and weighed, the weight of the livers was significantly increased in the HF group mice compared to the LC group, while the weight of the livers was significantly decreased after the intervention of diethylbutylmalonate (fig. 3).

3. Diethyl n-butylmalonate can reduce lipid deposition in liver tissue of mice

As shown in fig. 4, oil red O staining revealed significant vacuolar degeneration of liver and increased lipid deposition in HF group mice relative to LC mice; diethyl oxalate intervenes in the reduction of vacuolar degeneration and lipid deposition in obese mice.

4. Diethyl n-butylmalonate can improve oral glucose tolerance of mice

As shown in fig. 5, fasting blood glucose levels were significantly higher in the HF group mice than in the LC and HF + DM groups. After the mice are perfused with the glucose solution (2 g/kg), the blood glucose level of each group of mice reaches the highest after 30 min and then tends to decline, and the blood glucose level of the HF group is still obviously higher than that of the LC and HF + DM group by 120 min. Experimental data show that the HF mice have obvious glucose intolerance compared with the LC mice, and the diethyl n-butylmalonate can improve the glucose intolerance of the fat mice induced by high fat diet.

5. Diethyl n-butylmalonate can improve insulin resistance of mice

As shown in fig. 6, the insulin levels in the fasting serum of HF mice were significantly higher than in the LC group; fasting serum insulin levels were significantly reduced in HF + DM mice compared to HF. Calculating a steady-state model insulin resistance index (HOMA-IR), and finding that the HOMA-IR index of the HF mouse is obviously higher than that of the LC group; compared with the HF group, the HOMA-IR index of the mice in the HF + DM group is obviously reduced. The results show that diethyl n-butylmalonate can improve high-fat diet-induced insulin resistance.

6. Diethyl n-butylmalonate can promote mouse systemic fatty acid oxidation capacity

Figure 7 shows that HF mice have significantly reduced systemic fatty acid oxidation compared to the LC group; compared with the HF group, the oxidation capability of the fatty acid of the HF + DM group mice is obviously improved. This result suggests that diethyl n-butylmalonate can reduce lipid deposition in obese mice by enhancing fatty acid oxidation, promoting lipid breakdown or consumption.

7. Diethyl n-butylmalonate can increase energy consumption of mouse body

Figure 8 shows that HF mice consumed significantly less energy than the LC group; whereas the HF + DM group mice had a significant increase in energy expenditure compared to the HF group. The results show that the diethyl n-butylmalonate can reduce the body weight of the obese mouse by promoting thermogenesis, increasing energy consumption and consuming redundant energy of the body.

8. Diethyl n-butylmalonate did not affect energy intake in mice

Figure 9 shows that the average energy intake per HF mouse per day is significantly increased compared to the LC group; but energy intake was not significantly changed in the HF + DM group mice relative to the HF group. The results show that diethyl n-butylmalonate does not affect energy intake in obese mice.

In conclusion, the above embodiments show that diethyl n-butylmalonate can prevent lipid accumulation in liver tissues, increase fatty acid oxidation and energy consumption of mouse bodies, and further prevent and treat obesity; and can improve glucose tolerance and insulin resistance, and has effect in correcting obesity-related metabolic disorder.

The above disclosure is only for the specific embodiment of the present invention, but the embodiment of the present invention is not limited thereto, and any variations that can be made by those skilled in the art should fall within the scope of the present invention.