阅读说明：本技术 7-(2,2-二甲基-3-丁烯酰胺基)-八氢苯喹啉乙酸酯在制备降脂药物中的应用 (Application of 7- (2, 2-dimethyl-3-butenamido) -octahydro phenylquinoline acetate in preparation of lipid-lowering drugs ) 是由 玉万国 唐皎瑢 陈春华 于 2021-08-31 设计创作，主要内容包括：本发明属于单体化合物的用途技术领域,具体涉及7-(2,2-二甲基-3-丁烯酰胺基)-八氢苯喹啉乙酸酯在制备降脂和治疗肥胖症的药物中的用途。本发明通过体外、体内降脂实验模型研究发现,7-(2,2-二甲基-3-丁烯酰胺基)-八氢苯喹啉乙酸酯具有减少脂肪堆积和降血脂的活性,可用于制备降脂和治疗肥胖症的药物,适于大规模推广,具有潜在的应用价值。(The invention belongs to the technical field of application of monomer compounds, and particularly relates to application of 7- (2, 2-dimethyl-3-butenamido) -octahydro-phenylquinoline acetate in preparation of a medicament for reducing fat and treating obesity. According to the invention, through in vitro and in vivo lipid-lowering experimental model researches, the 7- (2, 2-dimethyl-3-butenamide) -octahydro-phenylquinoline acetate has the activities of reducing fat accumulation and reducing blood fat, can be used for preparing the lipid-lowering and obesity-treating medicines, is suitable for large-scale popularization, and has potential application value.)

Use of 7- (2, 2-dimethyl-3-butenamido) -octahydro-phenylquinoline acetate, i.e. 7- (2, 2-dimethyl-3-butenamido) -2,4-dimethyl-5-oxo-1,2,3,4,4a,5,6,10b-octahydrobenzo [ f ] -1-quinolyl-acetate, in the preparation of a medicament for the treatment of lipid lowering and obesity.

Technical Field

The invention belongs to the technical field of applications of monomer compounds, and particularly relates to an application of 7- (2, 2-dimethyl-3-butenamide) -octahydro-phenylquinoline acetate in preparation of a lipid-lowering drug.

Background

Hyperlipemia is one of common metabolic diseases in China, the incidence rate of hyperlipemia tends to increase year by year, and the hyperlipemia can cause a series of complications such as cardiovascular and cerebrovascular diseases and the like, and the life quality and the average life of the population in China are seriously influenced. Obesity is the main manifestation, and it is estimated that by 2050, global obesity will become a prevalent trend, with adult male obese patients rising to 65%, adult female obese patients rising to 50%, and minor children rising to around 25%. Survey data show that hyperlipidemia is an important factor induced by coronary heart disease, myocardial infarction, cerebral thrombosis and cerebral apoplexy, and the body is remarkably marked by abnormally high Triglyceride (TG) and Total Cholesterol (TC) levels. At present, lipid-lowering drugs commonly used in clinic are statins and fibrates, which have definite lipid-lowering effect and strong regulating effect on blood lipid, but are accompanied by certain toxic and side effects, such as muscle tissue damage, liver toxicity, abnormal digestive function (the concentration of cholesterol in bile is abnormally increased, and gallstone is caused), and the like. Safe, efficient and low-toxicity lipid-lowering active ingredients are searched in natural medicines, are concerned about and have potential application value.

7- (2, 2-dimethyl-3-butenamido) -octahydro-phenylquinoline acetate, i.e. 7- (2, 2-dimethyl-3-butenamido) -2,4-dimethyl-5-oxo-1,2,3,4,4a,5,6,10b-octahydrobenzo [ f]-1-quinolyl-acetate [7- (2, 2-dimethyllbut-3-enamido) -2,4-dimethyl-5-oxo-1,2,3,4,4a,5,6,10 b-octahydrobenzol [ f] quinolin-1-yl acetate, DQA]Molecular formula is C₂₃H₃₀N₂O₄The relative molecular weight is 398, and the molecular structure is shown below. Is mainly prepared by extracting and separating medicinal plant radix tinosporae japonicae endophyte.

At present, no related research report of 7- (2, 2-dimethyl-3-butenamide) -octahydro phenylquinoline acetate for lipid-lowering medicines exists.

Disclosure of Invention

The invention aims to solve the problems in the prior art and provides application of 7- (2, 2-dimethyl-3-butenamido) -octahydro-phenylquinoline acetate in preparing a medicament for reducing blood fat and treating obesity. The 7- (2, 2-dimethyl-3-butenamido) -octahydro phenylquinoline acetate can reduce the in vitro 3T3-L1 fat cell intracellular lipid accumulation, improve the content of extracellular glycerol, reduce the weight, visceral fat weight, blood fat and cholesterol level of mice induced by in vivo high-fat diet, can be used for preparing the medicines for reducing fat and treating obesity, and has potential application value.

In order to achieve the purpose, the invention has the following beneficial effects:

1. according to the invention, through research, the 7- (2, 2-dimethyl-3-butenamide) -octahydro phenylquinoline acetate can reduce in-vitro 3T3-L1 fat cell intracellular lipid accumulation and improve the content of extracellular glycerol, and can be used for preparing a medicament for reducing blood fat and treating obesity;

2. according to the invention, through research, the 7- (2, 2-dimethyl-3-butenamide) -octahydro phenylquinoline acetate can reduce the weight, visceral fat weight, triglyceride and cholesterol level in blood of a mouse induced by high-fat diet in vivo, and can be used for preparing medicines for reducing blood fat and treating hyperlipidemia.

Drawings

FIG. 1 is a bar graph of 7- (2, 2-dimethyl-3-butenamido) -octahydro-phenylquinoline acetate as a reducing lipid accumulation in 3T3-L1 adipocytes and increasing extracellular glycerol content, wherein: a is a lipid accumulation histogram and B is an extracellular glycerol content histogram; in panels a and B, the symbols indicate that there was a very significant difference compared to the blank control group (a)P<0.01), symbol indicates a significant difference compared to the blank control group (P<0.05）。

FIG. 2 is a bar graph of 7- (2, 2-dimethyl-3-butenamido) -octahydro-phenylquinoline acetate on a bar graph showing the reduction of body weight, visceral fat weight, triglycerides and cholesterol in blood of obese mice induced by a high fat diet, wherein: c is a bar graph of the body weight of the mouse, D is a bar graph of the visceral fat weight of the mouse, E is a bar graph of the content of triglyceride in the blood of the mouse, and F is a bar graph of the content of cholesterol in the blood of the mouse; in fig. 2, the symbol indicates that there was a very significant difference from the model control group (P<0.01), symbol indicates significant difference compared to model control group(s) ((ii)P<0.05), code # indicates significant difference compared to the blank control group: (P<0.05）。

Detailed Description

In order to clearly understand the technical contents of the present invention, the following examples are given in detail.

Example 1: 7- (2, 2-dimethyl-3-butenamido) -octahydro-phenylquinoline acetate, namely 7- (2, 2-dimethyl-3-butenamido) -2,4-dimethyl-5-oxo-1,2,3,4,4a,5,6,10b-octahydrobenzo [ f ] -1-quinolyl-acetate, has the activity test of reducing the fat of 3T3-L1 fat cells.

Reagents and drugs:

adipocytes (3T 3-L1, ATCC, Rockville, Md., USA) are able to store excess energy in the form of triglycerides, maintain a high differentiation capacity, can differentiate into mature adipoblasts by induction, and are an internationally accepted cell model for studying lipid metabolism; simvastatin (Sim), purchased from Sigma Aldrich Company (St. Louis, MO, USA) with a purity of >99.5% (HPLC); DQA, namely 7- (2, 2-dimethyl-3-crotonamide) -octahydro-phenylquinoline acetate, is extracted and separated from endophytes of aerial roots of the medicinal plant radix tinosporae by laboratories, and the purity is more than 98 percent (HPLC).

Experimental methods and results:

1.3T 3-L1 adipocyte differentiation and administration

Will be 1 × 10⁵3T3-L1 adipocytes at a density of one cell/well were seeded into 6-well plates and cultured for 48 h using DMEM medium containing 10% FBS. Then, the medium was replaced with fresh medium and DMEM (10% FBS, 0.5mM IBMX, 1. mu.M DEX, 10. mu.g/ml insulin) medium containing the inducer was added for differentiation for 96 h. Thereafter, differentiation medium (DMEM, 10% FBS, 10. mu.g/ml insulin) and various concentrations of DQA were added and the culture was continued for 24 hours. Simvastatin (Sim) is a positive drug.

2. Experimental results of DQA on adipocyte lipogenesis

As shown in figure 1, the series of concentrations of DQA can reduce intracellular lipid accumulation and present a dose-dependent relationship, wherein when the concentration of DQA is 5 μ M (as shown in figure 1 (A)), the effect is better than that of a positive control drug 5 μ M Sim. The DQA with the series of concentrations can improve the content of extracellular glycerol and present a dose-dependent relationship, wherein when the DQA concentration is 5 μ M (as shown in figure 1 (B)), the action effect is superior to that of a positive control drug 5 μ M Sim.

The experimental results show that the 7- (2, 2-dimethyl-3-butenamide) -octahydro-phenylquinoline acetate can reduce the in vitro lipid accumulation in 3T3-L1 fat cells and improve the content of extracellular glycerol, and can be used for preparing the medicines for reducing blood fat and treating obesity.

Example 2: 7- (2, 2-dimethyl-3-butenamide) -octahydro-phenylquinoline acetate, namely 7- (2, 2-dimethyl-3-butenamide) -2,4-dimethyl-5-oxo-1,2,3,4,4a,5,6,10b-octahydrobenzo [ f ] -1-quinolyl-acetate, is used for acute high fat induced fat mouse lipid accumulation and blood fat reduction experiments.

Reagents and drugs:

male C57BL/6 mice aged 4 weeks (lakan sleek seuda laboratory animals ltd); simvastatin (Sim), purchased from Sigma Aldrich Company (St. Louis, MO, USA) with a purity of >99.5% (HPLC); DQA, namely 7- (2, 2-dimethyl-3-crotonamide) -octahydro-phenylquinoline acetate, is extracted and separated from endophytes of aerial roots of the medicinal plant radix tinosporae by laboratories, and the purity is more than 98 percent (HPLC).

Experimental methods and results:

1. establishment of acute high fat induced obesity animal model

Male C57BL/6 mice at age 4 weeks were used for modeling. The purchased mice were randomly divided into 5 groups (blank control group, obesity model group, high dose group, low dose group, and positive drug control group) 10 mice each after being fed on a normal diet in the laboratory for one week. Mice in the placebo group were fed a low fat diet (10% fat, 14% protein, 76% carbohydrate, 3.6 kcal/g) and the remaining groups were fed a high fat diet (50% fat, 14% protein, 36% carbohydrate, 5.1 kcal/g) for 6 weeks to establish an acute high fat diet-induced mouse obesity model. The blank control group was injected intraperitoneally with 5% DMSO, and the remaining groups were injected intraperitoneally with DQA and positive drugs of different concentrations for 6 weeks.

2. Experimental result of DQA on lipid accumulation and lipid regulation of acute high-fat-induced obese mice

As shown in fig. 2, after the acute high-fat diet induced obesity modeling of mice, and the intraperitoneal injection administration treatment of the DQA with different concentrations, experimental results show that, compared with the obesity-induced model control group, the groups with different administration dosages of DQA can reduce the body weight of the mice (as shown in fig. 2 (C)), and show a dose-dependent relationship, wherein when the concentration of DQA is 20 mg/kg, the action effect is better than that of the positive control drug 20 mg/kg Sim; the weight of visceral fat of mice can be reduced by different administration dose groups of DQA (as shown in figure 2 (D)), and a dose-dependent relation is presented, wherein when the concentration of DQA is 20 mg/kg, the action effect is better than that of a positive control drug 20 mg/kg Sim; the level of triglyceride in the blood of mice can be reduced by different administration dose groups of the DQA (as shown in figure 2 (E)), and a dose-dependent relation is presented, wherein when the concentration of the DQA is 20 mg/kg, the action effect is better than that of a positive control drug 20 mg/kg Sim; the different administration dose groups of DQA can reduce the cholesterol level in the blood of mice (as shown in figure 2 (F)), and show a dose-dependent relationship, wherein when the concentration of DQA is 20 mg/kg, the effect is better than that of the positive control drug 20 mg/kg Sim.

From the above examples, it can be seen that 7- (2, 2-dimethyl-3-butenamido) -octahydro-phenylquinoline acetate can reduce the weight, visceral fat weight, triglyceride and cholesterol level in blood of mice induced by high fat diet in vivo, and can be used for preparing medicines for reducing blood lipid and treating hyperlipidemia.

In conclusion, the 7- (2, 2-dimethyl-3-butenamido) -octahydro-phenylquinoline acetate is derived from natural medicinal plant, namely the aerogenic rhizobium bruguinii, belongs to a pure natural source, is safe and reliable, has small toxic and side effects, can reduce the in vitro intracellular lipid accumulation of 3T3-L1 fat cells and improve the content of extracellular glycerol, and can be used for preparing medicines for reducing blood fat and treating obesity; can reduce the weight of mice induced by high-fat diet in vivo, the weight of visceral fat, and the level of triglyceride and cholesterol in blood, can be used for preparing medicines for reducing blood lipid and treating hyperlipidemia, and has potential application value.

In this specification, the invention has been described with reference to specific embodiments thereof. It will, however, be evident that various modifications and changes may be made thereto without departing from the broader spirit and scope of the invention. The specification and drawings are, accordingly, to be regarded in an illustrative rather than a restrictive sense.