A kind of dibenzofuran derivative and its preparation method and application

Abstract

The invention discloses a compound as shown in a formula I, and a stereisomer thereof, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof or a metabolite thereof. The invention provides a completely new compound as shown in the formula I, which has a good effect of rheumatoid arthritis resistance, and provides a novel approach for the clinical screening and / or preparation of rheumatoid arthritis resistance drugs.

Description

Technical field

[0001] The invention relates to a dibenzofuran derivative, and a preparation method and application thereof. Background technique

[0002] Bauhinia championi (Benth.) Benth is a woody vine of the genus Bauhinia of the leguminous family Bauhinia, aliased as Bauhinia vine, Wulang vine, Round dragon, Jiulong vine, scutellaria, plum blossom into bone, swallow tail Etc., distributed in Zhejiang, Jiangxi, Fujian, Guangdong, Guangxi, Hunan, Hubei, Guizhou and other provinces, it is flat, bitter, non-toxic, has the effects of dispelling wind, dehumidification, promoting qi and blood circulation, and is clinically used to treat rheumatic joints Inflammation, low back and leg pain, bruises, etc., the folks are mainly used to treat epigastric pain, rheumatic joint pain, acute and chronic low back and leg pain.

[0003] Although the Longxue vine plant has good curative effect, it has not been fully researched and developed. The further clarification of the composition of S. s...

Examples

Embodiment 1

[0084] Example 1 Preparation of the compound of the present invention

[0085] 1. Extract

[0086] (1) Take 63 kg of the rhizome of Rhizoma lanceolata, pulverize it into coarse powder, soak it with 120 L of 90% ethanol solution, and extract it by cold soaking for 3 times, each for 3 days. The extracts were combined and concentrated under reduced pressure to obtain 2650 g of Ethanol extract extract.

[0087] (2) Disperse and mix 2650 g of ethanol extract extract with 8 L of water, extract with petroleum ether and ethyl acetate in turn, concentrate under reduced pressure and vacuum dry to obtain ethyl acetate extract (435.0 g).

[0088] 2. Separation and purification of compounds

[0089] Take 435.0g of the ethyl acetate extract, use silica gel column chromatography (100-200 mesh) to separate and purify, mix and mix the samples, elution with chloroform, the gradient elution program is shown in Table 1

[0090] Table 1 Gradient elution program

[0091] Chloroform: Methanol (v / v) Elutio...

experiment example 1

[0107] Experimental Example 1 Anti-rheumatoid arthritis activity

[0108] The compound's proliferation inhibitory activity on synovial cells was detected by MTT method. Synovial cells were cultured in RPMI 1640 medium (containing 10% fetal bovine serum, 100μg / mL penicillin and streptomycin, 1mmol / L L-glutamine) at 37℃, 5% CO 2 Incubate in a saturated humidity incubator to the logarithmic growth phase. The synovial cells in the logarithmic growth phase were seeded into a 96-well plate (cell concentration of 1×105 / mL). After the cells adhere to the wall for 13 hours, add 100 μL of each of 9 test samples with different concentration gradients and the positive control drug methotrexate, and add an equal volume of administration medium to the negative control group. After co-incubating for 48 hours, add 10 μL of MTT solution (5 mg / mL) to each well, continue to incubate for 4 hours, and aspirate the supernatant. 100μL of DMSO was added, the crystals were fully dissolved, and the ligh...

Embodiment 2

[0110] Example 2 Pharmacodynamic study of anti-rheumatoid arthritis

[0111] Animals: SPF-grade healthy rats, Wistar, male, six weeks old, weighing 180-200g, purchased from Shanghai Slack Laboratory Animal Co., Ltd. [Certificate Number: SCXK (Shanghai) 2012-0002]. It is fed in an SPF laboratory at the Experimental Animal Center of Fujian University of Traditional Chinese Medicine. The light / dark cycle is 12 / 12h, the temperature is 25.0±0.5℃, and the relative humidity is 50%-70%.

[0112] 1. Establishment of a rat model of bovine collagen-induced arthritis (CIA)

[0113] Specific modeling method: mix bovine type II collagen (2mg / mL) with complete Freund's adjuvant (CFA) in equal volume, and grind to make a collagen emulsion. Inject 0.2 mL of collagen emulsion subcutaneously at the base of the rat’s tail 2 cm, and inject at the base of the rat’s tail 3 cm on the 7th day (a mixed emulsion of bovine type II collagen and incomplete Freund’s adjuvant (IFA)) 0.1mL / only for the second immu...

Technical field

[0001] The invention relates to a dibenzofuran derivative, and a preparation method and application thereof. Background technique

[0002] Bauhinia championi (Benth.) Benth is a woody vine of the genus Bauhinia of the leguminous family Bauhinia, aliased as Bauhinia vine, Wulang vine, Round dragon, Jiulong vine, scutellaria, plum blossom into bone, swallow tail Etc., distributed in Zhejiang, Jiangxi, Fujian, Guangdong, Guangxi, Hunan, Hubei, Guizhou and other provinces, it is flat, bitter, non-toxic, has the effects of dispelling wind, dehumidification, promoting qi and blood circulation, and is clinically used to treat rheumatic joints Inflammation, low back and leg pain, bruises, etc., the folks are mainly used to treat epigastric pain, rheumatic joint pain, acute and chronic low back and leg pain.

[0003] Although the Longxue vine plant has good curative effect, it has not been fully researched and developed. The further clarification of the composition of S. s...

Examples

Embodiment 1

[0084] Example 1 Preparation of the compound of the present invention

[0085] 1. Extract

[0086] (1) Take 63 kg of the rhizome of Rhizoma lanceolata, pulverize it into coarse powder, soak it with 120 L of 90% ethanol solution, and extract it by cold soaking for 3 times, each for 3 days. The extracts were combined and concentrated under reduced pressure to obtain 2650 g of Ethanol extract extract.

[0087] (2) Disperse and mix 2650 g of ethanol extract extract with 8 L of water, extract with petroleum ether and ethyl acetate in turn, concentrate under reduced pressure and vacuum dry to obtain ethyl acetate extract (435.0 g).

[0088] 2. Separation and purification of compounds

[0089] Take 435.0g of the ethyl acetate extract, use silica gel column chromatography (100-200 mesh) to separate and purify, mix and mix the samples, elution with chloroform, the gradient elution program is shown in Table 1

[0090] Table 1 Gradient elution program

[0091] Chloroform: Methanol (v / v) Elutio...

experiment example 1

[0107] Experimental Example 1 Anti-rheumatoid arthritis activity

[0108] The compound's proliferation inhibitory activity on synovial cells was detected by MTT method. Synovial cells were cultured in RPMI 1640 medium (containing 10% fetal bovine serum, 100μg / mL penicillin and streptomycin, 1mmol / L L-glutamine) at 37℃, 5% CO 2 Incubate in a saturated humidity incubator to the logarithmic growth phase. The synovial cells in the logarithmic growth phase were seeded into a 96-well plate (cell concentration of 1×105 / mL). After the cells adhere to the wall for 13 hours, add 100 μL of each of 9 test samples with different concentration gradients and the positive control drug methotrexate, and add an equal volume of administration medium to the negative control group. After co-incubating for 48 hours, add 10 μL of MTT solution (5 mg / mL) to each well, continue to incubate for 4 hours, and aspirate the supernatant. 100μL of DMSO was added, the crystals were fully dissolved, and the ligh...

Embodiment 2

[0110] Example 2 Pharmacodynamic study of anti-rheumatoid arthritis

[0111] Animals: SPF-grade healthy rats, Wistar, male, six weeks old, weighing 180-200g, purchased from Shanghai Slack Laboratory Animal Co., Ltd. [Certificate Number: SCXK (Shanghai) 2012-0002]. It is fed in an SPF laboratory at the Experimental Animal Center of Fujian University of Traditional Chinese Medicine. The light / dark cycle is 12 / 12h, the temperature is 25.0±0.5℃, and the relative humidity is 50%-70%.

[0112] 1. Establishment of a rat model of bovine collagen-induced arthritis (CIA)

[0113] Specific modeling method: mix bovine type II collagen (2mg / mL) with complete Freund's adjuvant (CFA) in equal volume, and grind to make a collagen emulsion. Inject 0.2 mL of collagen emulsion subcutaneously at the base of the rat’s tail 2 cm, and inject at the base of the rat’s tail 3 cm on the 7th day (a mixed emulsion of bovine type II collagen and incomplete Freund’s adjuvant (IFA)) 0.1mL / only for the second immu...

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