CN114736252B

CN114736252B - Forsythin derivative, preparation method and medical application thereof

Info

Publication number: CN114736252B
Application number: CN202210450591.7A
Authority: CN
Inventors: 沈庆坤; 郭红艳; 全哲山; 桑晓桐
Original assignee: Yanbian University
Current assignee: Yanbian University
Priority date: 2022-04-27
Filing date: 2022-04-27
Publication date: 2023-07-21
Anticipated expiration: 2042-04-27
Also published as: CN114736252A

Abstract

The invention discloses a forsythin derivative, a preparation method and medical application thereof, wherein the compound is a novel compound with IL-6 inhibition activity, and is structurally modified and optimized on the basis of natural product forsythin, so that the physicochemical property, pharmacological activity and patentability of the forsythin are improved; the invention also provides a preparation method of the forsythin derivative, which has the advantages of simple operation of the reaction process, mild reaction conditions, and cheap and easily available reagents; the novel phillyrin derivative effectively reduces the release of IL-6 by RAW264.7 cells induced by LPS, has the inhibition capability obviously superior to that of a lead compound phillyrin, can effectively inhibit macrophage inflammatory response induced by LPS, and is used in the field of anti-inflammatory drug preparation.

Description

Forsythin derivative, preparation method and medical application thereof

Technical Field

The invention discloses a forsythin derivative, which is a novel compound; the invention also provides a preparation method and medical application of the forsythin derivative, and belongs to the technical field of biological medicine.

Background

Human beings have long been exploring for inflammation, which is a common and frequent disease. The normal inflammatory response is a defensive process of the body and is a biological response of the immune system to harmful stimuli (e.g. viral, bacterial infections, toxins, toxic compounds, tissue damage). Usually acute inflammation can eliminate harmful stimulation and help to restore the normal health state of the body. However, uncontrolled acute inflammation may progress to chronic inflammation and may lead to various severe chronic diseases (e.g., tumors, various autoimmune diseases, alzheimer's disease, diabetes, cardiovascular disease, fibrosis, etc.). Although the pathogenesis of these diseases is different, most are related to the inflammatory pathway. To hinder the development of acute inflammation into persistent chronic inflammation, the inflammatory response must be suppressed to prevent additional tissue damage.

Currently, drugs are mainly used for treating inflammation in clinic, and common anti-inflammatory drugs are mainly divided Into Steroid (SAIDs) and Nonsteroidal (NSAIDs), and nonsteroidal anti-inflammatory drugs are the most widely used anti-inflammatory drugs in the world. Although they have powerful anti-inflammatory, analgesic and antipyretic activities, they have stronger toxic and side effects. Therefore, it is important to find anti-inflammatory drugs with low toxic and side effects.

Disclosure of Invention

The invention discloses a forsythin derivative, which is a novel compound, has stronger inflammatory factor IL-6 release inhibition capability, and can be applied to the anti-inflammatory field.

The invention discloses a preparation method of a forsythin derivative, which has the advantages of simple operation in the reaction process, mild reaction conditions, and cheap and easily available reagents.

The invention relates to a forsythin derivative, which has the following molecular formula: c (C) ₄₁ H ₅₁ NO ₁₄ The method comprises the steps of carrying out a first treatment on the surface of the The molecular weight is as follows: 781.33; the chemical structural formula is as follows:

the forsythin derivatives are named as follows:

((2R,3S,4S,5R,6R)-6-(4-((3aR,4R,6aR)-4-(3,4-dimethoxyphenyl)tetrahydro-1H,3H-furo[3,4-c]furan-1-yl)-2-methoxyphenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl(tert-butoxycarbonyl)-L-phenylalaninate; short for the sake of brevity: B5.

the invention discloses a preparation method of a forsythin derivative, which comprises the following specific preparation steps:

1. taking forsythin (0.08 mmol-0.12 mmol), BOC-L-phenylalanine (13 mmol-0.17 mmol), DMAP (08 mmol-0.12 mmol), EDC (0.18 mmol-0.22 mmol) and placing in a 25 mL round bottom flask, adding a 5 mL dichloromethane acetonitrile 1:1 mixed solution, and reacting for 12 hours under ice bath condition;

2. after the reaction was detected by thin layer chromatography, the solution was spin-dried, 10 mL distilled water was added,extracting with dichloromethane at least three times, mixing organic phases, and sequentially extracting with saturated NH ₄ Aqueous Cl solution, water, saturated aqueous NaCl solution and organic phase (saturated NH) ₄ The Cl aqueous solution removes residual DMAP), and anhydrous magnesium sulfate is added for drying, and the magnesium sulfate powder is removed by suction filtration and concentrated under reduced pressure to obtain a crude product.

3. Separating and purifying by silica gel column chromatography (dichloromethane: methanol=150:1-100:1, V/V) to obtain the target compound B5.

B5 nmr data as follows: m.p. 74-75 deg.C, yield 45%. ¹ H NMR (300 MHz, Chloroform-d) δ 7.40 – 7.31 (m, 2H), 7.24 (s, 1H), 7.18 (s, 2H), 7.16 (s, 1H), 6.96 (d, J = 8.1 Hz, 2H), 6.85 (d, J = 13.3 Hz, 3H), 4.98 (s, 1H), 4.89 (s, 1H), 4.68 (s, 2H), 4.44 (s, 3H), 4.12 (d, J = 9.0 Hz, 1H), 3.93 (s, 3H), 3.89 (d, J = 13.5 Hz, 6H), 3.85 (s, 2H), 3.57 (d, J = 27.3 Hz, 4H), 3.42 (s, 1H), 3.33 (d, J = 4.5 Hz, 2H), 3.12 (s, 2H), 2.85 (s, 2H), 1.44 (s, 9H). ¹³ C NMR (126 MHz, CDCl ₃ ) δ 171.57, 155.07, 150.26, 148.69, 147.86, 145.49, 137.16, 136.04, 130.93, 129.30(2C), 128.35(2C), 126.76, 118.26, 117.67, 111.03, 109.92, 108.95, 102.38, 87.20, 81.81, 79.58, 76.61, 74.01, 73.25, 70.90, 70.26, 69.55, 64.79, 60.22, 55.92, 55.83(2C), 54.32, 49.93, 29.17, 28.24(3C)。

The forsythin derivative has the application of IL-6 inhibition activity in the preparation of anti-inflammatory medicaments.

The invention has the positive effects that:

the compound has the advantages that the concentration dependence of RAW264.7 cells induced by LPS is reduced, the inhibition capability is obviously better than that of a lead compound, namely, the forsythin, can effectively inhibit macrophage inflammatory reaction induced by LPS, and is used in the field of anti-inflammatory drug preparation. The preparation method of the compound has the advantages of simple operation of the reaction process, mild reaction conditions, and cheap and easily available reagents.

Drawings

FIG. 1 is a graph showing the inhibitory effect of the phillyrin derivative B5 on the IL-6 release of RAW264.7 cells induced by LPS.

Detailed description of the preferred embodiments

The present invention will be further described with reference to examples, but the present invention is not limited to the above embodiments. Different embodiments may be realized by means of modifications within the scope of the claims, which modifications shall fall within the scope of the invention.

Example 1:

forsythin (0.08 mmol-0.12 mmol), BOC-L-phenylalanine (13 mmol-0.17 mmol), DMAP (08 mmol-0.12 mmol), EDC (0.18 mmol-0.22 mmol) are placed in a 25 mL round bottom flask, 5 mL dichloromethane acetonitrile 1:1 mixed solution is added, and the reaction is carried out for 12 hours under ice bath condition. Detecting the completion of the reaction by thin layer chromatography, spin-drying the solution, adding 10 mL distilled water, extracting with dichloromethane at least three times, mixing the organic phases, sequentially extracting with saturated NH ₄ The organic phase is washed by Cl water solution, water and saturated NaCl water solution, and is added with anhydrous magnesium sulfate for drying, and the coarse product is obtained after removing magnesium sulfate powder by suction filtration and concentrating under reduced pressure. Separating and purifying by silica gel column chromatography (dichloromethane: methanol=150:1-100:1, V/V) to obtain target compound B5; the molecular formula is: c (C) ₄₁ H ₅₁ NO ₁₄ The method comprises the steps of carrying out a first treatment on the surface of the The molecular weight is as follows: 781.33, the chemical structural formula is:

Example 2:

pharmaceutical composition: 1000 tablet formulations containing 100 mg active ingredient per tablet: 100 g the compound B5, 2 g hydroxypropyl methylcellulose, 10 g wheat starch, 100 g sucrose and 6 g magnesium stearate; the dosage employed should be adapted to the nature and severity of the disease, the route of administration, and the age and weight of the patient. Daily doses vary from 0.1 mg to 1.0 g and may be administered once or several times.

The medical use of the compounds of the invention is further demonstrated by the following tests:

test example 1

The effect of the lead compound forsythin (FOR, 25 μm) and various concentrations of compound B5 on the inhibition of RAW264.7 cells secretion of IL-6 was tested using an ELISA kit.

Experimental materials:

RAW264.7 mouse macrophages were purchased from China academy of sciences cell bank, DMEM (high sugar) cell culture broth was purchased from Sigma-Aldrich, fetal Bovine Serum (FBS) was purchased from Sigma-Aldrich, penicillin-streptomycin double antibody mixture was purchased from Bioind, murine IL-6 ELISA kit was purchased from eBioScience San Diego, and enzyme-labeled instrument was purchased from U.S. Thermo Scientific.

The experimental method comprises the following steps:

RAW264.7 mouse macrophages were seeded on 24-well plates at a density of 3X 10 ⁵ Up to 5X 10 ⁵ Cells/well 24-well plates inoculated with cells were placed in a cell incubator for 12 hours of incubation. The original culture medium was aspirated, the experimental group was added with DMEM medium in which different forsythin derivatives were dissolved, the remaining group was added with an equal volume of blank DMEM medium, after pretreatment for 1 hour, the model group and the experimental group were added with DMEM medium containing LPS (100 ng/mL), and the blank group was given the same volume of blank DMEM medium, incubated for 24 hours, culture supernatants were collected and the IL-6 content was determined and calculated by ELISA method, step-wise procedure with reference to the kit instructions, see FIG. 1.

Experimental results:

compound B5 significantly reduced LPS-induced release of IL-6 by RAW264.7 cells and had a certain concentration dependence. The inhibition capacity of compound B5 is significantly enhanced compared to the lead compound Forsythin (FOR) at the same concentration (25 μm). It is demonstrated that compound B5 can effectively inhibit LPS-induced macrophage inflammatory response.

Claims

1. A forsythin derivative, characterized by having the following structural formula:

；

the molecular formula is: c (C) ₄₁ H ₅₁ NO ₁₄ The method comprises the steps of carrying out a first treatment on the surface of the Molecular weight: 781.33.

2. the preparation method of the forsythin derivative according to claim 1, comprising the following steps:

1) Taking 0.08-0.12 mmol of forsythin, 0.13-0.17 mmol of BOC-L-phenylalanine, 0.08-0.12 mmol of DMAP and 0.18-0.22 mmol of EDC, placing the mixture in a 25 mL round bottom flask, adding a 5 mL dichloromethane acetonitrile 1:1 mixed solution, and reacting for 12 hours under ice bath condition;

2) After the reaction was completed, the mixture was extracted three times with dichloromethane, and the organic phases were combined, followed by saturated NH ₄ The organic phase is washed with aqueous Cl, water, saturated aqueous NaCl and dried with anhydrous magnesium sulfate, the solids are removed by suction filtration and concentrated under reduced pressure, and the mixture is taken through dichloromethane: methanol=150: 1-100:1, and separating and purifying by silica gel column chromatography of V/V.

3. Use of a forsythin derivative according to claim 1 for the preparation of an anti-inflammatory drug having IL-6 inhibitory activity.

4. A pharmaceutical formulation comprising a forsythin derivative according to claim 1 as active ingredient together with one or more pharmaceutically acceptable adjuvants.