CN114099493A - Active compound for inhibiting insulin resistance and application thereof - Google Patents
Active compound for inhibiting insulin resistance and application thereof Download PDFInfo
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- A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
- A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin
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Abstract
The invention relates to a compound for inhibiting insulin resistance activity and application thereof, wherein the compound is selected from one or a combination of two of hyperin and 5, 7, 3 ', 4' -tetrahydroxy flavanol. The hyperin and 5, 7, 3 ', 4' -tetrahydroxyflavanol of the present invention have an insulin resistance inhibitory activity, and particularly when they are combined in a specific ratio, they contribute to the synergistic insulin resistance inhibitory activity. The invention has important significance for providing a novel medicine for treating type II diabetes.
Description
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 (IR) refers to a state in which the biological effect of Insulin in blood is reduced, i.e., the body's responsiveness to the biological regulation of Insulin is reduced, and the effect of Insulin in promoting glucose uptake is impaired, resulting in increased compensatory Insulin secretion, which is marked by hyperinsulinemia, specifically, decreased sensitivity of peripheral tissues to Insulin and impaired glucose utilization. Insulin resistance is the common pathophysiological basis of many diseases of metabolic syndrome, such as obesity, hyperglycemia, type 2 diabetes, hyperlipidemia, coronary heart disease, hypertension, atherosclerotic coronary heart disease, etc. According to statistics, the number of obese people in the world exceeds 10 hundred million, and more than 80 percent of obese patients show obvious IR; while the population with type 2 diabetes has also reached 1.5 billion, the incidence of IR in type 2 diabetic patients is as high as around 85%. It is further reported that a pregnant woman has a 50% decrease in insulin sensitivity and is likely to develop insulin resistance. Therefore, how to relieve insulin resistance and improve insulin sensitivity is an important subject for preventing and treating obesity, type 2 diabetes, hypertension, hyperlipidemia, atherosclerotic coronary heart disease and other diseases.
The molecular mechanism of insulin resistance generation has not yet been fully elucidated, and numerous studies have demonstrated that insulin resistance occurs in close correlation with phosphatidylinositol-3-hydroxykinase (PI 3-hydroxykinase, PKB) insulin signaling pathway blockade, abnormal glucose transporter4 (GLUT 4) translocation, glycogen synthesis Kinase-3 β (GSK-3 β) activation, decreased extracellular signal-regulated Kinase activity, and the like. The PI3K/PKB pathway is the major pathway by which insulin exerts hypoglycemic effects. The clinical treatment of IR is still directed to the control of its unilateral symptoms, mainly insulin sensitizers, e.g. thiazolidinediones, increase the glucose utilization in peripheral tissues. The preparation method is matched with hypoglycemic drugs aiming at hyperglycemia, antihypertensive drugs aiming at hypertension and the like, so that the preparation method is complicated in link, high in cost, great in side effect and free of simple and effective comprehensive prevention and treatment means. Moreover, the western medicine can generate tolerance, and the dosage of the medicine is increased continuously until the medicine is ineffective. The search for simple and effective multi-target omnibearing treatment methods is urgent. Despite the last half century of research, there are still too many unknowns on the pathogenesis of EMS. Drug and surgical therapy remain the current primary treatment, but relapse remains a problem that is confusing to the industry. Therefore, deep analysis of the pathogenesis of the internal abnormality lays a foundation for better understanding of the disease and finding more effective treatment measures in the industry, and has important strategic significance for improving the health quality of women and the health harmony of the whole family and the society.
The hypericum attenuatum lindl is also called as steady heart grass and rhizome of Mount bustle, is a perennial herb, is 30-70 cm high, and has developed lateral roots and fibrous roots. It has bitter taste and mild nature, and has effects of stopping bleeding, relieving pain, promoting lactation, etc., and is mainly suitable for hemoptysis, hematemesis, metrorrhagia, rheumatic arthralgia, neuralgia, traumatic injury, galactostasis, mastitis; it is used externally to treat traumatic hemorrhage, carbuncle, furuncle and pyogenic infections. In recent years, scholars at home and abroad make great progress on the research on chemical components and pharmacological actions of hypericum attenuatum.
The inventors of the present application have also long worked on the study of Hypericum attenuatum and published various academic papers, e.g., Wang Yan et al, "the Effect of Total Flavonoids of Hypericum attenuatum on the rectified potassium current in the rat models of cardiac arrhythmias," Journal of Chinese medicine (2016.). further, for example, Wang Yan et al, "infection of Total flavone from Hypericum erectum Choil on induced rectifier granule current of cardiac tissue complex of China" Journal of Chinese medicine 031.002(2016):656 ti. further, for example, Feng Y, Teng L, Wang Y, use of spectral-Effective derivatives of tissue with LC-to-Screen-MS-tissue J. the existence of pancreatic polypeptide J. the Japanese aspic. also has been reported that the inhibitory activity of hypericin J. A. the inventors of Hypericum attenuatum Extracts of Hypericum attenuatum by Japanese et al, have reported that the inhibitory activity of pancreatic epithelial cells of pancreatic cell J. A. The present inventors have unexpectedly found an active compound capable of inhibiting insulin resistance through studies on active ingredients in hypericum attenuatum, and have completed the present invention based on this finding.
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:
one aspect of the present 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 compound is prepared as a pharmaceutically acceptable formulation.
In another preferred embodiment of the present invention, the pharmaceutically acceptable formulation is an oral formulation, e.g., a tablet, a capsule, a liquid formulation, a sustained release formulation.
In another preferred embodiment of the present invention, the pharmaceutically acceptable formulation further comprises pharmaceutically acceptable excipients.
Another aspect of the present invention relates to a composition for inhibiting insulin resistance activity, comprising hyperin and 5, 7, 3 ', 4' -tetrahydroxyflavanol as active ingredients, wherein the weight ratio of hyperin to 5, 7, 3 ', 4' -tetrahydroxyflavanol is 1: 4-3: 2; preferably 1.8 to 2.2: 2.8-3.2. According to the invention, the hyperin and the 5, 7, 3 ', 4' -tetrahydroxy flavanol are combined in a specific ratio, so that the synergistic effect of the hyperin and the 5, 7, 3 ', 4' -tetrahydroxy flavanol on inhibiting the insulin resistance activity is exerted.
In another aspect, the invention relates to the use of the above active compound or pharmaceutical composition 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 5, 7, 3 ', 4' -tetrahydroxyflavanol of the present invention have an insulin resistance inhibitory activity, and particularly when they are combined in a specific ratio, they contribute to the synergistic insulin resistance inhibitory activity. 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, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Unless otherwise specified, the reagents involved in the examples of the present invention are all commercially available products, and all of them are commercially available.
Example 1
1. Material
1.1 Hyperoside (hereinafter referred to as Compound 1) and 5, 7, 3 ', 4' -tetrahydroxyflavanol (hereinafter referred to as Compound 2) were separated and purified by chromatography according to literature methods (Dongjiayong et al, research on chemical components of flavonoids in Verbena stringtchuensis [ J ]. J. Med. 2005, 40(12):897-899.)
1.2 Experimental animals
200 SPF-grade female SD rats of 3 weeks old were bred in the animal center of the first hospital affiliated to the university of traditional Chinese medicine in black dragon river. Feeding at constant temperature (50% humidity) of 25 deg.C for clean level, without feeding vitamin product, and periodically alternating 12h light and 12h dark.
2 method of experiment
2.1 establishment of IR rat model
An insulin resistance rat model was established by feeding high-fat high-sugar diet. Feed formulation (g/100 g): fructose 60%, fat 15%, protein 21%, fiber 3%, vitamins and minerals 1%.
2.2 rat grouping and dosing methods
After the rats are successfully molded, excluding the rats which are not successfully molded, and randomly dividing the rats into: the test group was a model group, a positive control group (rosiglitazone 1mg/kg/d), and test groups 1 to 5 (administered after being administered in physiological saline at the ratio shown in Table 1), and the normal control group was fed with 20 normal feeds. The total number of the groups is 8, and each group contains 20. The experimental group is administrated by gastric lavage, the dosage of each gastric lavage is 1mg/kg/d, 1 time per day, and the duration is 8 weeks. During the experiment rats had free access to water and food intake was recorded every 1d and body weight was recorded every 3 days. After the last feeding, fasting is not forbidden for 10h, the heart is bled, and serum is separated conventionally and stored at 4 ℃ for later use.
2.3 Observation index
(1) Determination of fasting blood glucose level
After the last feeding, the rats were fasted for 5h without water deprivation, 10. mu.L of blood was taken from the tail vein and added to 0.2mL of protein precipitant, and left to stand at room temperature for 7min, 4500r/min, centrifuged for 15min, and 150. mu.L of supernatant was taken and fasting blood glucose level was determined according to the method described in the glucose kit.
(2) Determination of oral glucose tolerance
After the last feeding, the rats are fasted for 5 hours without water prohibition, the rats are subjected to intragastric administration of 2g/kg of glucose solution, 10 mu L of blood is taken from the tail vein after intragastric administration for 0, 0.5, 1 and 2 hours respectively to determine the blood glucose level at the moment, and the area under the glucose curve (AUC) is calculated according to the following formula:
AUC (h mmol/L) of 0.25 xa +0.5 xb +0.75 xc +0.5 xd (A, B, C, D indicates blood glucose levels of 0, 0.5, 1, 2h, respectively)
(3) Determination of serum insulin levels
After the last feeding, the rats are fasted for 10 hours without water prohibition, the eyeballs are picked, blood is taken, after standing for 30min at room temperature, the blood is centrifuged for 15min at 7500r/min, 10 mu L of blood serum is taken, and the blood serum insulin level is measured according to the specification of an ELISA kit. The insulin resistance index (HOMA-IR) and the insulin sensitivity detection index (QUICKI) were respectively calculated from the following formulas:
HOMA-IR fasting blood glucose level x serum insulin level/22.5
QUICKI 1/(lg fasting blood glucose level + lg serum insulin level)
(4) Determination of body weight gain in rats
2.4 statistical analysis:
the experimental results are expressed as x + -s, and single factor one-way ANOVA analysis of variance is performed by SPSS 22.0 software, and two-by-two comparison is performed by Tukeys method, and P <0.05 is used as the difference with statistical significance.
3 results of the experiment
3.1 general conditions in rats
The normal control group rats have glossy fur, good mental state performance, sensitive action response and active behaviors.
After the model group rats are successfully modeled, the fur is dry and lusterless, the mental state is poor, the action is slow, the response is slow, the activity is poor, and the degree of the activity is aggravated along with the raising time.
After the drug modeling is finished, the gavage is started for drug treatment, and compared with a model group, the behavioral performance of rats in each experimental group is improved to a certain extent, wherein the experimental group 4 is most obvious.
3.2 weight gain in rats
TABLE 1 Effect of active Compounds on insulin resistance in rats weight gain (x. + -. s)
Group of | N | Administration (total 1mg/kg/d) | Food intake (g) | Weight gain (g) |
Normal group | 20 | - | 285±16 | 105±16 |
Positive control group | 20 | Rosiglitazone | 226±18# | 167±14* |
Model set | 20 | - | 229±20# | 192±15## |
Experimental group 1 | 20 | Compound 1 | 218±17# | 172±19* |
Experimental group 2 | 20 | Compound 2 | 225±19# | 183±20* |
Experimental group 3 | 20 | Compound 1: compound 2 is 1: 4 | 217±15# | 152±15* |
Experimental group 4 | 20 | Compound 1: compound 2 is 2: 3 | 215±19# | 132±17** |
Experimental group 5 | 20 | Compound 1: compound 2 is 3: 3 | 223±21# | 149±16* |
Note: # P <0.05, # P <0.01, compared to the normal control group. P <0.05, P <0.01 was compared to model groups.
3.3 Effect of active Compounds on insulin resistance in rat-related indices
Table 2 effect of active compound on various indices 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 set | 13.48±1.24## | 14.7±0.4## | 8.32±0.72## | 0.382±0.015## |
Experimental group 1 | 10.38±1.34* | 13.4±0.6* | 5.95±0.58* | 0.456±0.012* |
Experimental group 2 | 10.52±1.27* | 13.8±0.7* | 5.87±0.49* | 0.445±0.014* |
Experimental group 3 | 9.98±1.30** | 12.3±0.7* | 4.63±0.47* | 0.491±0.015* |
Experimental group 4 | 9.66±1.02** | 11.5±0.8* | 4.56±0.57* | 0.501±0.011* |
Experimental group 5 | 10.12±1.22** | 12.1±0.9* | 5.11±0.61* | 0.478±0.018* |
Note: # P <0.01 compared to normal control group; p <0.05 was compared to model groups.
From the experimental results in table 2, it can be seen that the fasting blood glucose levels of the model rats were significantly higher than those of the normal control group (P < 0.01). The fasting blood glucose level of rats of each group of the active compound is obviously reduced compared with that of the model control group; most significant in experimental group 4. No statistical significance compared to the positive control group indicates that experimental group 4 has been able to achieve similar activity to rosiglitazone. The results show that each group of the experiment can obviously reduce the fasting blood glucose level of RI rats, and the experiment group 4 has the best effect. As can be seen from the experimental results in table 2, each group of the experiment had significant effects in reducing insulin levels, reducing insulin resistance, and increasing insulin sensitivity, and the group 4 (hyperin: 7, 3 ', 4' -tetrahydroxyflavanol ═ 2: 3) had the best effect.
The foregoing describes preferred embodiments of the present invention, but is not intended to limit the invention thereto. Modifications and variations of the embodiments disclosed herein may be made by those skilled in the art without departing from the scope and spirit of the invention.
Claims (10)
1. A compound for inhibiting insulin resistance activity, which is selected from one or a combination of two of hyperin and 5, 7, 3 ', 4' -tetrahydroxy flavanol.
2. The compound of claim 1, prepared in a pharmaceutically acceptable formulation.
3. The compound of claim 2, wherein the pharmaceutically acceptable formulation is an oral formulation.
4. The compound of claim 3, which is a tablet, capsule, liquid formulation, or sustained release formulation.
5. The compound of claim 2, wherein the pharmaceutically acceptable formulation further comprises a pharmaceutically acceptable excipient.
6. A composition for inhibiting insulin resistance activity comprising as active ingredients hyperin and 5, 7, 3 ', 4' -tetrahydroxyflavanol, wherein the weight ratio of hyperin to 5, 7, 3 ', 4' -tetrahydroxyflavanol is 1: 4-3: 2; preferably 1.8 to 2.2: 2.8-3.2.
7. The composition of claim 6, prepared in a pharmaceutically acceptable formulation.
8. The compound of claim 7, wherein the pharmaceutically acceptable formulation is an oral formulation.
9. The application of the active compound or the pharmaceutical composition refers to the application in preparing the medicines for inhibiting insulin resistance.
10. The use according to claim 9, wherein the medicament is for the treatment of type II diabetes.
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