阅读说明：本技术 橘皮素在制备抑制β分泌酶活性药物中的应用 (Application of hesperetin in preparation of drugs for inhibiting beta-secretase activity ) 是由 舒细记 包建 梁征 龚晓康 柳威 孙宾莲 于 2020-12-11 设计创作，主要内容包括：本发明公开了橘皮素在制备抑制β分泌酶活性药物中的应用,同时本发明还公开了橘皮素在制备用于治疗由β分泌酶活性增强所致的疾病的药物中的应用。本发明首次发掘了橘皮素的作用靶点为β分泌酶,本发明发现并验证了橘皮素具有与β分泌酶抑制剂相似的结合β分泌酶的能力,橘皮素通过与β分泌酶结合并抑制其活性,减少了β分泌酶对淀粉样蛋白前体蛋白的错误剪切,进而降低了脑内淀粉样蛋白Aβ的产生和错误堆积,从而达到改善阿尔兹海默症,增强学习记忆能力的作用。并且由于橘皮素具有抑制β分泌酶活性的作用,因此任何由于β分泌酶活性增强导致的疾病均可通过橘皮素进行改善和治疗。(The invention discloses application of hesperetin in preparation of a drug for inhibiting beta secretase activity, and also discloses application of hesperetin in preparation of a drug for treating diseases caused by enhancement of beta secretase activity. The invention discovers and verifies that the hesperetin has the ability of combining beta secretase similar to that of a beta secretase inhibitor, and the hesperetin is combined with the beta secretase and inhibits the activity of the beta secretase, so that the error shearing of the beta secretase on amyloid precursor protein is reduced, the generation and error accumulation of amyloid A beta in brain are further reduced, and the effects of improving Alzheimer's disease and enhancing learning and memory capacity are achieved. And any disease caused by the enhancement of beta-secretase activity can be improved and treated by the hesperetin because the hesperetin has the effect of inhibiting the beta-secretase activity.)

1. Application of hesperetin in preparation of drugs for inhibiting beta-secretase activity.

2. Use of hesperetin for the preparation of a medicament for the treatment of a disease resulting from an enhanced beta-secretase activity.

3. The use according to claim 2, wherein the disease is alzheimer's disease.

4. The use according to claim 1 or 2, wherein the medicament is in any pharmaceutically acceptable form.

5. The use according to claim 4, wherein the dosage form is a tablet, granule, capsule, oral liquid or pill.

6. Use according to claim 1 or 2, wherein the hesperetin is prepared as a milk powder, a beverage or a solid beverage.

Technical Field

The invention relates to the technical field of medical health products, in particular to application of hesperetin in preparation of a drug for inhibiting beta secretase activity.

Background

Alzheimer's Disease (AD), also known as senile dementia, is a highly harmful senile disease. According to statistics, the incidence of the disease is increasing year by year in China. With the aging of our society, alzheimer's disease will bring huge medical and financial expenses and social burden. Therefore, the development of drugs capable of alleviating or treating the disease has been a hot point of research.

The current mainstream theory holds that one of the main causes of alzheimer's disease is the wrong accumulation of amyloid a β in the brain, resulting in neurotoxic senile plaques, eventually leading to neuronal death. In the brain, Amyloid Precursor Protein (APP) is mis-cleaved by beta-secretase (BACE-1) and gamma-secretase to produce Amyloid A β, where BACE-1 cleavage of APP is the rate-limiting step in this pathway. Therefore, to interfere with amyloid A β production, one major research focus has been on limiting the activity of BACE-1. However, several drugs known so far, such as BACE-1 inhibitors, virally-entrapped BACE1-siRNA, etc., generally have unthinkable side effects, such as severe retinopathy or other safety issues. At present, no proper safe specific medicine is available at home and abroad.

A series of secondary metabolite plant compounds (phytochemicals) derived from plants have been proved to have good pharmacological functions in recent years, and particularly, flavonoid substances derived from edible fruits and vegetables have the characteristics of good drug effect and low toxic and side effects, so that the phytochemicals are widely concerned and developed into commodities such as medicines, dietary supplements and the like. Among them, the polymethoxyflavonoids of hesperetin found in citrus fruits have been confirmed to have excellent pharmacological actions such as oxidation resistance, inflammation resistance, liver protection, nerve protection, etc., and the prior art also discloses that hesperetin has an action of preventing dementia of the alzheimer type. However, no research on the action target of the hesperetin on the Alzheimer disease exists at present.

Disclosure of Invention

The invention aims to provide application of hesperetin in preparation of a drug for inhibiting activity of beta-secretase, and the invention discovers that the beta-secretase (BACE-1) is a molecular target of the hesperetin for the first time, namely the invention discovers and verifies that the hesperetin can be combined with the BACE-1 and inhibit the activity of the BACE-1, so that the generation of a mistaken shearing product amyloid A beta of the hesperetin is reduced, the mistaken accumulation of the amyloid A beta in brain is reduced, and the effects of improving Alzheimer's disease and enhancing learning and memory ability are achieved.

In order to achieve the purpose, the invention adopts the technical scheme that:

the invention provides application of hesperetin in preparation of a drug for inhibiting beta secretase activity.

The invention also provides the application of the hesperetin in preparing a medicament for treating diseases caused by the enhancement of the activity of beta-secretase.

Further, the disease is alzheimer's disease.

Further, the dosage form of the medicine is any pharmaceutically acceptable dosage form.

Furthermore, the dosage form is tablets, granules, capsules, oral liquid or pills.

Further, the tablet comprises the following components in parts by weight: 55 parts of hesperetin, 44 parts of anhydrous lactose and 1 part of magnesium stearate.

Further, the granules comprise the following components in parts by weight: 75 parts of hesperetin and 25 parts of anhydrous lactose.

Further, the hesperetin is prepared into milk powder, beverage or solid beverage.

Further, when the hesperetin is prepared into milk powder, the milk powder comprises the following components in parts by weight: 20 parts of hesperetin, 70 parts of skim milk powder, 5 parts of xylitol and 5 parts of erythritol.

Further, when the hesperetin is prepared into a solid beverage, the solid beverage comprises the following components in parts by weight: 30 parts of hesperetin, 30 parts of skim milk powder, 10 parts of antarctic krill oil powder, 10 parts of inulin, 10 parts of microcrystalline cellulose, 5 parts of xylitol and 5 parts of erythritol.

Compared with the prior art, the invention has the beneficial effects that: the invention discovers and verifies that the hesperetin has the ability of combining beta secretase similar to that of a beta secretase inhibitor, and the hesperetin is combined with the beta secretase and inhibits the activity of the beta secretase, so that the error shearing of the beta secretase on amyloid precursor protein is reduced, the generation and error accumulation of amyloid A beta in brain are further reduced, and the effects of improving Alzheimer's disease and enhancing learning and memory capacity are achieved. And any disease caused by the enhancement of beta-secretase activity can be improved and treated by the hesperetin because the hesperetin has the effect of inhibiting the beta-secretase activity.

Drawings

FIG. 1 is a graph showing the results of molecular docking simulation of hesperetin and BACE1-IN-1 with the known inhibitor of beta-secretase activity BACE1 IN example 1 of the present invention, wherein A is the best docking result of hesperetin and BACE-1, and B is the best docking result of BACE-IN-1 and BACE-1;

FIG. 2 is a graph showing the detection results of the escape latency of mice in Morris water maze training for 5 days in example 2 of the present invention;

FIG. 3 is a result of measuring the number of times that a mouse passes through a platform region in a space exploration experiment, when the Morris water maze is trained for 6 days in example 2 of the present invention;

FIG. 4 is a graph showing the results of detecting the expression level of s-APP β protein in mouse brain tissue in example 2 of the present invention, wherein A is the result of immunoblot detection and B is the result of quantitative detection of the expression level of s-APP β protein.

Detailed Description

The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the 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.

EXAMPLE 1 inhibition of BACE-1 Activity by hesperetin

This example compares the binding ability of an hesperetin molecule (available from Douguerie scientific development Co., Ltd.) and a known chemically synthesized BACE-1 inhibitor BACE1-IN-1 (available from Shanghai ceramic Biotechnology Co., Ltd.) to BACE1 by molecular docking simulation, and is specifically performed as follows:

BACE-1 molecules (PDB ID: 2WJO) had small molecules (QUD) and all water molecules bound to them removed by PyMOL Molecular Graphics System (2.4.0) software, and had the charge equally distributed and hydrogen atoms added by Autodock tools 4.2. Structural documents of both hesperetin and BACE-IN-1 molecules were downloaded IN the NCBI database. The software used for molecular docking is autodock4.2, the docking algorithm is a ramak genetic algorithm, and the range of molecular docking is the overall structure of BACE-1.

Docking results are shown IN FIG. 1, A is the best docking mode of hesperetin and BACE-1, and B is the best docking mode of BACE-IN-1 and BACE-1. According to the results, both the hesperetin and the BACE-IN-1 can be docked with the BACE-1, namely the hesperetin has the similar ability of binding BACE-1 molecules with a BACE-1 inhibitor and has approximate binding energy, wherein the binding energy of the hesperetin and the BACE-1 is-5.82 kcal/mol, and the binding energy of the BACE-IN-1 and the BACE-1 is-6.24 kcal/mol. The molecular structure of the hesperetin is proved to have the potential of inhibiting BACE-1.

Example 2 improvement of learning and memory ability and inhibition of BACE-1 Activity of hesperetin in APP/PS1 mice: an Alzheimer's disease transgenic mouse model, namely an APP/PS1 double transgenic mouse.

The experimental method comprises the following steps: 12 February half-aged APP/PS1 mice were selected and divided into two groups on average, the administration group (Tangeretin group) was orally intragastrically administered with 100mg/kg hesperetin (dissolved in 1% sodium carboxymethylcellulose aqueous solution) per day, and the model group (APP/PS1 group) was orally intragastrically administered with 1% sodium carboxymethylcellulose aqueous solution of the same volume per day. Four months later, two groups of animals were evaluated for learning and memory levels using the Morris water maze experiment: wherein, when the Morris water maze is trained for 5 days, the detection result of the escape latency of the mouse is shown in figure 2; the results of the detection of the number of times the mouse crossed the platform region in the space exploration experiment when the Morris water maze was trained for 6 days are shown in FIG. 3. The animals were then sacrificed and brain homogenates were taken to measure the expression level of s-APP β protein in the brain tissues of mice in the administration group and model group, and the results are shown in FIG. 4.

The APP/PS1 transgenic mouse selected in the experiment is a double-transgenic animal and can express mutant human presenilin (DeltaE9) and human Amyloid Precursor Protein (APP). The study reports that at six months of age, APP/PS1 mice have A beta deposition in the brain, and have a reduced level of learning and memory caused by A beta deposition. The Morris water maze is a classical behavioral experiment used to evaluate the level of learning and memory in animals.

According to the test results of fig. 2, on the 5 th day of training, mice of the administration group (tageretin) had escape latency significantly shorter than those of the model group (APP/PS1) (p < 0.05); from the results of the examination shown in FIG. 3, in the space exploration experiment performed on the 6 th day of training, the mice of the administration group (Tageretin) found the platform region more easily than the mice of the model group (APP/PS 1). The results in fig. 2 and fig. 3 demonstrate that APP/PS1 mice have significantly improved learning and memory abilities when administered orally with hesperetin.

The immunoblot and s-APP β protein expression quantitative determination results shown in FIG. 4 show that the expression level of s-APP β protein in the brain homogenate of the mice in the administration group (APP/PS1+ Tan) is very significantly reduced (p <0.01) compared with the brain homogenate of the model group (APP/PS1), wherein the s-APP β protein is the protein residue after BACE-1 cleaves APP, which indicates that the activity of BACE-1 is significantly inhibited after the mice take hesperetin orally, thereby reducing the generation of BACE-1 mistaken cleavage product amyloid A β and reducing the mistaken accumulation of amyloid A β in brain, and further treating and improving Alzheimer's disease.

Example 3

The hesperetin can be prepared into different dosage forms or products to prepare medicaments for inhibiting the activity of beta-secretase or medicaments for treating diseases caused by the enhancement of the activity of the beta-secretase.

The hesperetin can be prepared into tablets, which comprise the following components in parts by weight: 55 parts of hesperetin, 44 parts of anhydrous lactose and 1 part of magnesium stearate;

the hesperetin can be prepared into granules, which comprise the following components in parts by weight: 75 parts of hesperetin and 25 parts of anhydrous lactose.

The hesperetin can be prepared into milk powder, which comprises the following components in parts by weight: 20 parts of hesperetin, 70 parts of skim milk powder, 5 parts of xylitol and 5 parts of erythritol.

The hesperetin can be prepared into solid beverage, which comprises the following components in parts by weight: 30 parts of hesperetin, 30 parts of skim milk powder, 10 parts of antarctic krill oil powder, 10 parts of inulin, 10 parts of microcrystalline cellulose, 5 parts of xylitol and 5 parts of erythritol.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention.