阅读说明：本技术 具有抗补体活性的苯并吡喃类化合物及其用途 (benzopyran compounds with anti-complement activity and application thereof ) 是由 周微 叶超 孙金凤 李镐 马英杰 王荣申 于 2019-09-25 设计创作，主要内容包括：本发明公开了一种具有抗补体活性的苯并吡喃类化合物及其在制备抗补体药物中的用途。本发明利用天然药物化学、现代药理学研究方法,对杜鹃花科杜鹃花属植物兴安杜鹃的抗补体活性物质进行研究,从95％乙醇提取物的乙酸乙酯萃取物中分离得到两种苯并吡喃类化合物,本发明的苯并吡喃类化合物对免疫补体经典途径具有抑制作用,其抑制作用CH<Sub>50</Sub>值为0.90-1.87mM,可用于制备补体抑制剂。(The invention discloses a benzopyran compound with anticomplementary activity and application thereof in preparing anticomplementary drugs. The invention utilizes the research method of natural medicinal chemistry and modern pharmacology to research the anticomplementary active substance of rhododendron dauricum of rhododendron in Ericaceae, two benzopyran compounds are separated from the ethyl acetate extract of 95 percent ethanol extract, the benzopyran compounds have the inhibiting effect on the classical pathway of immune complement, and the inhibiting effect CH of the benzopyran compounds 50 The value is 0.90-1.87mM, and can be used for preparing complement inhibitor.)

1. Benzopyran compounds with anticomplementary activity have a structural general formula shown as a formula I:

In the formula I, the compound has the following structure,

when R ═ a, the compound is daurichromene E;

When R ═ B, the compound is daurichromene a.

2. The process for producing benzopyran-based compounds according to claim 1, comprising the steps of:

(1) Soaking dried Rhododendron dauricum leaf in 95% ethanol to obtain Rhododendron dauricum leaf ethanol extract;

(2) suspending the ethanol extract of the rhododendron dauricum leaves obtained in the step (1) with distilled water, and sequentially extracting with petroleum ether, ethyl acetate and n-butanol respectively to obtain a petroleum ether extract, an ethyl acetate extract and a n-butanol extract;

(3) And (3) separating and purifying the ethyl acetate extract obtained in the step (2) by using a positive and negative phase silica gel column chromatography and a medium and low pressure liquid chromatography to obtain a compound daurichromene E and a compound daurichromene A.

3. The method according to claim 2, wherein in the step (1), the dried Rhododendron dauricum leaves are extracted by soaking with 95% ethanol at room temperature for 7 days for 3 times to obtain Rhododendron dauricum leaf ethanol extract.

4. Use of the benzopyran-based compound according to claim 1 in the preparation of anticomplementary drugs.

5. the use according to claim 4, wherein the benzopyran-based compound inhibits the classical pathway of the complement system.

6. Use according to claim 5, characterized in that the benzopyran compounds inhibit the classical pathway of the complement system CH₅₀the value is 0.90-1.87 mM.

Technical Field

The invention belongs to the field of traditional Chinese medicine natural medicine pharmacy, and relates to benzopyran compounds with an anti-complement effect and application thereof.

Background

The complement system is part of the human innate immune system and plays an important role in homeostasis, inflammation, and innate defense against foreign pathogens. Over-activation of the complement system in cells can be detrimental and is associated with a range of diseases including inflammatory diseases, viral and bacterial infections and gastrointestinal diseases. Thus, modulation of complement activity can be beneficial in the treatment of various complement-associated diseases. Activation of the complement system is composed primarily of three distinct pathways. The Classical Pathway (CP) is activated by the formation of immune complexes on the pathogen surface and calreticulin expressed on apoptotic cells, which results in the binding of the C1 complex. The Lectin Pathway (LP) recognizes mannose-terminating glycans on pathogens, which lead to activation of the MBL MASP complex. Alternative Pathways (AP) are permanently active at low levels to investigate the presence of pathogens in healthy individuals. Of these three pathways, the classical pathway is considered to be an important pathway for the prevention and treatment of diseases.

Complement inhibitors are a very active field of new drug development in recent years, and clinically used for treating diseases caused by excessive activation of complement system include small molecules, polypeptides, proteins, antibodies/novel antibodies, aptamers, spiegelmers, and the like. Although the development of complement inhibitor drugs is very active, many challenges are faced, and the currently marketed complement inhibitors cannot meet the requirements of high efficiency, low toxicity and specificity. The natural plant has the characteristics of complex components, low cost, low toxicity and the like, so that the screening of the complement inhibitor meeting the clinical requirement becomes possible.

Rhododendron dauricum is a semi-evergreen shrub belonging to genus Rhododendron of Ericaceae, and is mainly distributed in northeast, Mongolia, Korea, Japan and Russia of China. Research on the phytochemistry of rhododendron dauricum shows that the rhododendron dauricum is rich in terpenoids, flavonoids, phenols and other components, and shows various biological activities including anti-HIV, anti-oxidation, antibacterial, analgesic and anti-inflammatory effects, neuroprotective activity, vasodilation and myocardial protection.

at present, the research of rhododendron dauricum leaves on the aspect of anticomplement activity is not reported, and the extraction and purification of rhododendron dauricum leaves and the application of rhododendron dauricum leaves in the fields of health care products, foods, medicines and the like are in an exploration stage.

Disclosure of Invention

the invention aims to provide a compound which is derived from natural plants and has an anticomplementary effect and application thereof. The invention extracts and separates two benzopyran compounds from rhododendron dauricum leaves, and has good anticomplementary effect.

In a first aspect, the present invention provides a compound represented by formula i, or a stereoisomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof:

(A)(B)

In the formula I, the compound has the following structure,

when R ═ a, the compound is daurichromene E, known in chinese as azachromene E;

When R ═ B, the compound is daurichromene a, referred to in chinese as azachromene a.

The second aspect of the present invention also provides a preparation method of the benzopyran compound, comprising the following steps:

(1) Soaking dried Rhododendron dauricum leaf in 95% ethanol to obtain Rhododendron dauricum leaf ethanol extract;

(2) Suspending the ethanol extract of the rhododendron dauricum leaves obtained in the step (1) with distilled water, and sequentially extracting with petroleum ether, ethyl acetate and n-butanol respectively to obtain a petroleum ether extract, an ethyl acetate extract and a n-butanol extract;

(3) And (3) separating and purifying the ethyl acetate extract obtained in the step (2) by using a positive and negative phase silica gel column chromatography and a medium and low pressure liquid chromatography to obtain a compound daurichromene E and a compound daurichromene A.

In the technical scheme, in the step (1), the dried rhododendron dauricum leaves are soaked and extracted by 95% ethanol at room temperature for 3 times, and the ethanol extract of the rhododendron dauricum leaves is obtained after 7 days each time.

the third aspect of the invention provides the application of the benzopyran compound in preparing anti-complement medicines. The benzopyran compound has an inhibition effect on the classical pathway of the complement system, and the benzopyran compound has an inhibition effect CH on the classical pathway of the complement system₅₀The value is 0.90-1.87 mM.

the invention has the beneficial effects that:

The invention provides benzopyran compounds shown as a formula I, which are monomeric compounds obtained by separating and purifying dried leaves of rhododendron, and have good anticomplementary activity. The pharmacological experiment result shows that the compound has an inhibiting effect on the classical pathway of immune complement, and the inhibiting effect CH₅₀The value is 0.90-1.87 mM. The compound can be used for preparing complement inhibitors and further preparing anticomplement medicines.

the invention extracts and separates two benzopyran compounds with anticomplementary function from rhododendron dauricum leaves for the first time, and has important practical significance and wide market prospect for comprehensively developing rhododendron dauricum resources and mining anticomplementary active ingredients in rhododendron dauricum leaves. At present, reports about the application of the benzopyran compounds to anticomplementary drugs are not found, and the new application of the anticomplementary activity belongs to the discovery for the first time.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. Unless otherwise specified, the experimental methods adopted by the invention are all conventional methods, and experimental equipment, materials, reagents and the like used in the experimental method can be purchased from chemical companies.

unless otherwise specified, the structural formulae depicted herein are intended to include all isomeric forms (e.g., enantiomers, diastereomers and geometric isomers (or conformers): e.g., the R, S configuration containing an asymmetric center, (Z), (E) isomers of double bonds, etc..) and, thus, individual stereochemical isomers of the compounds of the present invention or mixtures of enantiomers, diastereomers or geometric isomers (or conformers) thereof are intended to be within the scope of the present invention.

The term "solvate" as used herein refers to a complex of a compound of the present invention coordinated to solvent molecules in a specific ratio.

The term "hydrate" as used herein refers to a complex formed by the coordination of the compound of the present invention with water.

As used herein, "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention with an acid or base, which is suitable for use as a pharmaceutical. Pharmaceutically acceptable salts, including inorganic and organic salts.

materials used in the following examples:

Xingan rhododendron leaf: collected leaves of Rhododendron dauricum L in extended area of Jilin province, and naturally dried to obtain the final product.