CN111632052A - Application of piperine in the preparation of sEH inhibitor - Google Patents

Abstract

The invention relates to the technical fields of traditional Chinese medicine chemistry and molecular biology, and discloses the application of piperine in the preparation of sEH inhibitors. Alkali has a good inhibitory effect on sEH activity. The compound has less toxic and side effects and good water solubility. Based on the action principle of soluble epoxide hydrolase, the compound has the effect of preventing and treating digestive tract diseases such as peptic ulcer, and can be used for preparing sEH inhibitor.

Description

Application of piperine in the preparation of sEH inhibitor

technical field

The invention relates to the technical fields of traditional Chinese medicine chemistry and molecular biology, in particular to the application of piperine isolated from pepper in the preparation of sEH inhibitors.

Background technique

At present, peptic ulcer disease (PUD), a digestive tract disease, has become a common public health problem in my country. Peptic ulcer is a common multiple disease with an incidence of about 4% worldwide (NajmW I.Peptic ulcer disease[J].Primary Care Clinics in Office Practice,2011,38(3):383-394.) , can occur at different ages, duodenal ulcers are more common in adolescents, gastric ulcers are more common in middle-aged and elderly people (Chen Haozhu, Lin Guowei, etc. Practical Internal Medicine [M]. Shanghai: Fudan University Press, 2010: 1981.) . Statistics show that in 2015, about 87.4 million people were diagnosed with this disease worldwide, and 267,500 people died (Collaborators G 2MM. Global, regional, and national levels of maternal mortality, 1990-2015: asystematic analysis for the Global Burden of Maternal mortality). Disease Study

2015[J]. Lancet, 2016, 388(10053):1775-1812.). Although the mortality rate of PUD is not high compared with other diseases, it will bring great inconvenience to the life and labor of patients, and it is impossible for any treatment plan to achieve 100% eradication (Investigation Collaborative Group. China Digestive Investigation report on the status quo of ulcer treatment [J]. Chinese Journal of Digestive Medicine, 2007, 27(2). Studies have shown that if chronic ulcers are not treated in time, the mucosal epithelium at the edge of the ulcer will gradually develop into atypical hyperplasia due to long-term stimulation, resulting in cancer. Although considerable research achievements have been made in the diagnosis and treatment of digestive tract diseases, the discovery of new treatment approaches and drugs for such diseases is still a major focus of current research.

Soluble epoxide hydrolase (sEH) is an enzyme that plays an irreplaceable role in mammals in nature. It is characterized by high selectivity to lipid epoxide fatty acids. Studies have shown that sEH mainly exists in the liver, kidney, gut, brain and vasculature in mammals. Epoxyeicosatrienoic acids (EETs) are the biologically active metabolites of arachidonic acid, and sEH can be used as a highly efficient catalyst in mammals to catalyze the metabolic inactivation of EETs and convert them into corresponding reactants. of dihydroxyeicosatrienoic acids (DHETs). EETs with powerful biological activity can participate in the process of a variety of physiological functions. They can dilate blood vessels, lower blood pressure and other functions in mammals. Anti-inflammatory effect in disease.

In this principle, by inhibiting sEH enzyme activity and increasing the level and activity of EETs in the body, peptic ulcer, chronic gastritis and other digestive tract diseases can be alleviated and treated. Because of this, the search for potentially effective sEH inhibitors as medicines for the treatment of hypertension, pain, and inflammatory diseases has become a hot research topic. Xu Danyan of Central South University in my country and Professor Long Yaqiu of Shanghai Institute of Materia Medica, Chinese Academy of Sciences are shifting their research focus to the research and development of sEH inhibitors. Foreign companies such as Merck have successively developed four types of sEH inhibitors, and Pfizer and Japan's Sumitomo Pharmaceutical have also made progress in this field. Nowadays, the search for new and effective sEH inhibitors has undoubtedly become an important direction for the research and development of new drugs for digestive tract diseases and anti-inflammatory and analgesic.

Pepper is a traditional Chinese medicine with a long history of medicinal use. It was first recorded in "Xinxiu Materia Medica", "Pepper produces Xirong, which is shaped like a buckthorn, and it tastes very spicy." Pepper is spicy, hot in nature, and returns to the stomach and large intestine meridians. effect. Modern medical research shows that pepper can be used to treat symptoms such as indigestion, cough, bronchitis, abdominal pain and diarrhea. Piperine is a compound extracted from pepper and its main active ingredient. Piperine has antioxidant, sedative, anticonvulsant, hypolipidemic, antidepressant, antitumor and anti-inflammatory effects on gastrointestinal diseases in biomedicine. In recent years, the biological activities and pharmacological effects of piperine have been widely studied, but most of them are studies on digestive tract diseases and various types of inflammation from the perspective of immunology. So far, there is no study on it as a soluble epoxy compound hydrolase inhibitor. and reports.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide the application of piperine in the preparation of sEH inhibitor.

The technical scheme of the present invention is summarized as follows:

Application of piperine in the preparation of sEH inhibitor, described piperine is represented by formula (I),

The sEH inhibitor piperine can treat peptic ulcer.

Advantages of the present invention: the results of enzyme kinetic activity test and pharmacophore show that piperine has a better inhibitory effect on sEH activity. The compound has few side effects and good water solubility. Based on the action principle of soluble epoxide hydrolase, the compound has the effect of preventing and treating digestive tract diseases such as peptic ulcer, and can be used for preparing sEH inhibitor.

Piperine can be prepared into a pharmaceutical composition with a pharmaceutically acceptable carrier to act as a sEH inhibitor.

Detailed ways

The present invention will be further described below through specific embodiments.

Example 1

Inhibitory activity test of piperine on sEH:

Piperine (purity ≥98%) was purchased from D-chem (Shanghai Xushuo Biotechnology Co., Ltd.)

The specific substrate (3-phenyl-oxy)-acetic acid cyano-(6-methoxy-naphthalen-2-yl) methyl ester, namely PHOME (Cayman Chemical Company, Ann Arbor, Michigan) itself has no fluorescence, But under the action of sEH, the product 6-methoxy-2-naphthaldehyde was hydrolyzed to produce 6-methoxy-2-naphthaldehyde, which can emit fluorescence with a wavelength of 465 nm under the excitation of 330 nm light wave. The intensity of the detected fluorescent signal was inversely proportional to the intensity of the inhibitory effect of sEH. According to the above principles, the enzymatic activity of sEH was determined by the reaction rate method. On the reaction curve, select the segment closest to the straight line in the initial stage, and then calculate the initial reaction velocity Vmax (indicating the enzyme activity level). The Vmax of the enzyme after treatment of each sample was calculated, and then compared with the normal control to calculate the percent inhibition rate of each sample. The kinetic monitoring mode continuously reads the fluorescence signal intensity value at each time point, which represents the production amount of the product. According to the enzyme reaction rate method, the production rate of the product can reflect the activity of the enzyme.

Piperine was dissolved in dimethyl sulfoxide at final concentrations of 0.1, 1, 10, 100.0 μM. All compounds were hydrolyzed with sEH by adding PHOME in 96-well plates at 37°C, and then the fluorescence signal was detected at 465nm. The probe substrate population (without evaluation compound) was used as a control. TPPU, a soluble cyclooxygenase inhibitor, was used as a control drug.

Kinetic analysis of the sEH enzyme of piperine was carried out by double reciprocal plot and Dixon technique. This assay was conducted in a range of PHOME concentrations of 1-30 [mu]M in positive and negative control experiments.

Download the x-ray crystal structures of soluble epoxide hydrolase (sEH) and its cognate ligands from the Protein Data Bank (PDB) ( http://www.rcsb.org/pdb/home/home.do , PDB ID: 4OCZ ). The 3D structure of piperine was constructed and minimized by Discovery Studio 3.5 (Biovia, San Diego, CA, USA). Subsequently, the docking of receptors and ligands was performed using the CDOCKER protocol in Discovery Studio 3.5z. The compound piperine can be well docked into the catalytic site cavity of sEH. However, no hydrogen bonds were formed between the compound and protein residues. This compound forms a π-bond interaction with residue His524, which is Asp 335-Asp 496-His524 (aspartate 335-aspartate 496-histidine 524) in the sEH cyclohydrolase domain ) part of the catalytic ternary domain.

The measured test results are as follows:

Table 1. Inhibitory effect of piperine on sEH;

Table 2. Kinetic parameters of piperine on sEH;

Table 3. Information on the interaction of piperine with sEH.

Table 1. Inhibitory effect of piperine on sEH

Table 2. Kinetic parameters of piperine on sEH

Table 3. Interaction information of piperine and sEH

Activity test results showed that the compound inhibited sEH activity.

Claims (3)

1. the application of piperine in the preparation of sEH inhibitor, described piperine is shown in formula (I):

2. The application according to claim 1, wherein the sEH inhibitor can be used to prepare medicines for the treatment of hypertension, pain and inflammatory diseases. 3. The application of piperine as sEH inhibitor in the preparation of medicine for treating peptic ulcer.