CN114099493B - Active compound for inhibiting insulin resistance and application thereof - Google Patents

Abstract

The invention relates to a compound for inhibiting insulin resistance activity and application thereof, wherein the compound is selected from one or two of hyperin and 5,7,3',4' -tetrahydroxy flavanol. The hyperin and the 5,7,3',4' -tetrahydroxyflavanol have the action of inhibiting the insulin resistance, and particularly when the hyperin and the 5,7,3',4' -tetrahydroxyflavanol are combined in a specific proportion, the synergistic action of inhibiting the insulin resistance is exerted. The invention has important significance for providing a novel medicine for treating type II diabetes.

Description

Active compound for inhibiting insulin resistance and application thereof

Technical Field

The invention belongs to the technical field of medical biology, and particularly relates to an active compound for inhibiting insulin resistance and application thereof.

Background

Insulin resistance (Insulin resistance, IR) refers to a state in which the biological effect of insulin in blood is reduced, that is, the body's responsiveness to the biological regulation of insulin is reduced, and the insulin-promoting action of glucose uptake is impaired, resulting in an increase in compensatory insulin secretion, an important sign of which is hyperinsulinemia, and particularly, a decrease in insulin sensitivity to peripheral tissues and a disorder of glucose utilization. Insulin resistance is a common pathophysiological basis for a number of diseases of the metabolic syndrome, such as obesity, hyperglycemia, type 2 diabetes, hyperlipidemia, coronary heart disease, hypertension, atherosclerotic coronary heart disease, etc. It is counted that the number of obese people in the world is over 10 hundred million, and more than 80% of obese patients show obvious IR; the population suffering from type 2 diabetes has reached 1.5 hundred million, and the incidence rate of IR in type 2 diabetes patients is as high as about 85%. It has been reported that pregnant women have 50% reduced insulin sensitivity and are susceptible to insulin resistance. Therefore, how to alleviate insulin resistance and improve insulin sensitivity has become an important topic for preventing and treating diseases such as obesity, type 2 diabetes, hypertension, hyperlipidemia, atherosclerosis coronary heart disease and the like.

The molecular mechanism of insulin resistance generation has not been fully elucidated at present, and a large number of studies have demonstrated that the occurrence of insulin resistance is closely related to phosphatidylinositol 3-hydroxykinase (PI 3K)/Protein Kinase B (PKB) insulin signaling pathway blockage, abnormal translocation of glucose transporter4 (glucose transporter, GLUT 4), glycogen synthesis Kinase-3 beta (glycogen synthase Kinase-3 beta, GSK-3 beta) activation, reduction in extracellular signal-regulating Kinase activity, and the like. The PI3K/PKB pathway is the primary pathway by which insulin exerts hypoglycemic effects. Clinically IR treatment is still directed to control of unilateral symptoms, with insulin sensitizer treatment being the primary, e.g. thiazolidinediones increase glucose utilization in peripheral tissues. The traditional Chinese medicine is matched with hypoglycemic drugs aiming at hyperglycemia or antihypertensive drugs aiming at hypertension and the like, has complicated links, high cost and large side effect, and has no simple and effective comprehensive prevention and treatment means. Moreover, western medicines can produce tolerance, and the dosage of the medicine is continuously increased until the medicine fails. Searching for a simple, effective, multi-target, and all-round therapeutic approach is an urgent need. Although studied for nearly half a century, there are too many undiagnosed puzzles on the pathogenesis of EMS. Drug and surgical therapies remain the current mainstay of therapy, but recurrence is still a problem that is confusing in the industry. Therefore, the deep analysis of the pathogenesis of the internal disorder lays a foundation for better understanding the disease in the industry and finding more effective treatment measures, and has important strategic significance for improving the health quality of women and the health harmony of the whole family and society.

The hypericum japonicum is also called heart-stabilizing grass and has the effect of expelling the penis, is perennial herbaceous plant, and has the height of 30-70 cm, and developed lateral roots and fibrous roots. The Chinese medicinal composition has the effects of stopping bleeding, easing pain, promoting lactation and the like, is mainly suitable for hemoptysis, hematemesis, uterine bleeding, rheumatic arthralgia, neuralgia, traumatic injury, milk deficiency and mastitis; it is indicated for traumatic hemorrhage, carbuncle and furuncle. In recent years, scholars at home and abroad have made great progress in studying chemical components and pharmacological actions of hypericum japonicum.

The inventors of the present application have also made long efforts for the study of hypericum and published a number of academic papers, for example, wang Yanli et al, "influence of hypericum total flavone on inward rectifying potassium current in rat myocardial cell arrhythmia model" journal of Chinese medicine (2016),. For example, wang Yanli, et al, "Influence of total flavonoids from Hypericum attenuatum Choisy on inward rectifier potassium current of rat cardiac muscle cell arrhythmia model," journal of Chinese medicine 031.002 (2016): 656-658. For example, feng Y, teng L, wang Y, et al, using Spectrum-Effect Relationships Coupled with LC-TOF-MS to Screen Anti-arrhythmic Components of the Total Flavonoids in Hypericum attenuatum Extracts [ J ]. Journal of Chromatographic Science,2020. However, the inventors of the present application have unexpectedly found an active compound capable of inhibiting insulin resistance through the study of active ingredients in hypericum.

Disclosure of Invention

In order to inhibit insulin resistance, the invention provides a compound for inhibiting insulin resistance activity and application thereof. Specifically, the invention adopts the following technical scheme:

in one aspect, the invention relates to a compound that inhibits insulin resistance activity selected from one or a combination of two of hyperin and 5,7,3',4' -tetrahydroxyflavanol.

In a preferred embodiment of the invention, the compounds are prepared as pharmaceutically acceptable formulations.

In another preferred embodiment of the invention, the pharmaceutically acceptable formulation is an oral formulation, e.g. a tablet, a capsule, a liquid formulation, a slow release formulation.

In another preferred embodiment of the present invention, the pharmaceutically acceptable formulation further comprises pharmaceutically acceptable excipients.

In another aspect, the present invention relates to a composition for inhibiting insulin resistance activity comprising hyperin and 5,7,3',4' -tetrahydroxy flavanol as active ingredients, wherein the weight ratio of hyperin to 5,7,3',4' -tetrahydroxy flavanol is 1: 4-3: 2; preferably 1.8-2.2:2.8-3.2. According to the invention, hyperin and 5,7,3',4' -tetrahydroxy flavanol are combined in a specific proportion, so that the synergistic effect of inhibiting insulin resistance activity of the hyperin and the 5,7,3',4' -tetrahydroxy flavanol can be exerted.

In a further aspect the invention relates to the use of the above-mentioned active compounds or pharmaceutical compositions, said use being in the manufacture of a medicament for inhibiting insulin resistance.

In a preferred embodiment of the invention, the medicament is for the treatment of type II diabetes.

Advantageous effects

The hyperin and the 5,7,3',4' -tetrahydroxyflavanol have the action of inhibiting the insulin resistance, and particularly when the hyperin and the 5,7,3',4' -tetrahydroxyflavanol are combined in a specific proportion, the synergistic action of inhibiting the insulin resistance is exerted. The invention has important significance for providing a novel medicine for treating type II diabetes.

Detailed Description

In order to further understand the present invention, a technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

Unless otherwise specified, all reagents involved in the examples of the present invention are commercially available products and are commercially available.

Example 1

1. Material

1.1 hyperin (hereinafter referred to as Compound 1) and 5,7,3',4' -tetrahydroxyflavanol (hereinafter referred to as Compound 2) were isolated and purified by chromatography according to the literature method (Dong Jianyong et al, J. Chinese J. Pharmaceutical, 2005, 40 (12): 897-899.)

1.2 laboratory animals

200 SPF-class SD rats of 3 weeks old were bred in the first hospital animal center affiliated to the university of Heilongjiang traditional Chinese medicine. The cleaning-grade feeding is carried out at the constant temperature of 25 ℃ (50% humidity), the vitamin product is not fed, and 12 hours of illumination and 12 hours of darkness are periodically and alternately carried out.

2 Experimental methods

2.1 construction of IR rat model

An insulin resistant rat model is established by a method of feeding high-fat and high-sugar feed. Feed formula (g/100 g): 60% of fructose, 15% of fat, 21% of protein, 3% of fiber, and 1% of vitamins and minerals.

2.2 rat grouping and administration methods

After successful molding, the rats without successful molding are removed and randomly divided into: the model group, the positive control group (rosiglitazone 1 mg/kg/d), the experimental group 1-5 groups (which were administered by being placed in physiological saline according to the ratio shown in Table 1), and the normal control group were fed with 20 conventional feeds. A total of 8 groups of 20. The experimental group was given by gavage, each time at 1mg/kg/d, 1 time a day for 8 weeks. Rats were fed free of water during the experiment, food intake was recorded every 1d, and body weight was recorded every 3 days. After the last feeding, the patients are fasted without water forbidding for 10 hours, hearts are taken, serum is conventionally separated, and the serum is stored at 4 ℃ for standby.

2.3 observations index

(1) Determination of fasting blood glucose levels

After the last feeding, the rats are fasted without water inhibition for 5 hours, 10 mu L of tail vein blood is taken and added into 0.2mL of protein precipitant, standing is carried out for 7min at room temperature, centrifugation is carried out for 15min, 150 mu L of supernatant is taken, and the fasting blood glucose level is measured by referring to a glucose kit instruction method.

(2) Determination of oral glucose tolerance

After the last feeding, rats are fasted without water inhibition for 5 hours, 2g/kg of glucose solution is infused, 10 mu L of tail vein blood is respectively taken after 0, 0.5, 1 and 2 hours of gastric infusion, the blood glucose level at the moment is measured, and the area under a glucose curve (areaunder the curve, AUC) is calculated according to the following formula:

AUC (h.mmol/L) =0.25×A+0.5×B+0.75×C+0.5×D (A, B, C, D represents blood glucose levels of 0, 0.5, 1, 2h, respectively)

(3) Determination of serum insulin levels

After the last feeding, the rats are fasted without water inhibition for 10 hours, eyeballs are picked up to take blood, the rats are kept stand at room temperature for 30 minutes, and are centrifuged for 15 minutes at 7500r/min, 10 mu L of serum is taken, and the serum insulin level is measured by referring to ELISA kit instructions. The model evaluation insulin resistance index (insulin resistance index, HOMA-IR) and insulin sensitivity detection index (quantitative insulin sensitivity check index, QUICKI) were calculated according to the following formulas, respectively:

HOMA-IR = fasting blood glucose level x serum insulin level/22.5

Quacki=1/(lg fasting blood glucose level+lg serum insulin level)

(4) Rat weight gain determination

2.4 statistical analysis:

the experimental results are expressed as x+ -s, and SPSS 22.0 software is used for one-way ANOVA variance analysis, and Tukey's method is used for two-by-two comparison, and P <0.05 is used as the difference with statistical significance.

3 results of experiments

3.1 general status of rats

The normal control group rats had glossy fur, good mental state, sensitive action response and active behavior.

After the model group rats are successfully modeled, the fur is dry and matt, the mental state is poor, the action is slow, the reaction is slow, the activity is poor, and the degree of the activity is aggravated along with the feeding time.

After the drug composition is molded, the stomach is infused to perform drug treatment, and compared with a model group, the behavior of rats in each experimental group is improved to a certain extent, wherein the experimental group 4 is the most remarkable.

3.2 weight gain of rats

TABLE 1 Effect of active Compounds on weight gain of insulin resistant rats (x+ -s)

Group of	N	Administration (1 mg/kg/d total)	Food intake (g)	Weight gain (g)
					Normal group	20	-	285±16	105±16
Positive control group	20	Rosiglitazone	226±18#	167±14*
					Model group	20	-	229±20#	192±15##
Experiment group 1	20	Compound 1	218±17#	172±19*
					Experiment group 2	20	Compound 2	225±19#	183±20*
Experiment group 3	20	Compound 1: compound 2 is 1:4	217±15#	152±15*
					Experiment group 4	20	Compound 1: compound 2 is 2:3	215±19#	132±17**
Experiment group 5	20	Compound 1: compound 2 is 3:3	223±21#	149±16*

Note that: #P <0.05, #P <0.01, compared to the normal control group. * P <0.05, < P <0.01, compared to model group.

3.3 Effect of active Compounds on insulin resistant rat related index

Table 2 effect of active compounds on various indicators of insulin resistant rats (n=20)

Group of	Fasting blood glucose (mmol/L)	Serum insulin (mU/L)	HOMA-IR	QUICK1
					Normal group	9.32±1.52	10.9±0.5	4.56±0.42	0.523±0.009
Positive control group	10.27±1.40**	13.2±0.8*	5.41±0.47*	0.442±0.012*
					Model group	13.48±1.24##	14.7±0.4##	8.32±0.72##	0.382±0.015##
Experiment group 1	10.38±1.34*	13.4±0.6*	5.95±0.58*	0.456±0.012*
					Experiment group 2	10.52±1.27*	13.8±0.7*	5.87±0.49*	0.445±0.014*
Experiment group 3	9.98±1.30**	12.3±0.7*	4.63±0.47*	0.491±0.015*
					Experiment group 4	9.66±1.02**	11.5±0.8*	4.56±0.57*	0.501±0.011*
Experiment group 5	10.12±1.22**	12.1±0.9*	5.11±0.61*	0.478±0.018*

Note that: # P <0.01 compared to the normal control group; * P <0.05 was compared to the model group.

As can be seen from the experimental results in Table 2, the fasting blood glucose level of the model rats was significantly higher than that of the normal control group (P < 0.01). The fasting blood glucose level of rats in each group of active compounds is significantly reduced compared with the model control group; most notably in experimental group 4. The comparison with the positive control group was not statistically significant, indicating that experimental group 4 was already able to achieve an activity similar to rosiglitazone. The results show that each experimental group can obviously reduce the fasting blood glucose level of the RI rat, and the experimental group 4 has the best effect. From the experimental results in table 2, each experimental group had remarkable effects in reducing insulin level, reducing insulin resistance, and increasing insulin sensitivity, and experimental group 4 (hyperin: 7,3',4' -tetrahydroxyflavanol=2:3) was most effective.

The foregoing describes preferred embodiments of the present invention, but is not intended to limit the invention thereto. Modifications and variations to the embodiments disclosed herein may be made by those skilled in the art without departing from the scope and spirit of the invention.

Claims (3)

1. Use of a composition for inhibiting insulin resistance activity, said composition consisting of hyperin and 5,7,3',4' -tetrahydroxyflavanol in a weight ratio of hyperin to 5,7,3',4' -tetrahydroxyflavanol of 1.8-2.2, for the preparation of a medicament for inhibiting the treatment of type two diabetes mellitus: 2.8-3.2.

2. The use according to claim 1, wherein the medicament is prepared as a pharmaceutically acceptable formulation.

3. The use according to claim 2, wherein the pharmaceutically acceptable formulation is an oral formulation.