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CN-117924401-B - Compound with neuroinflammation inhibition activity and extraction method thereof

CN117924401BCN 117924401 BCN117924401 BCN 117924401BCN-117924401-B

Abstract

The invention relates to the field of medicines, in particular to a compound with neuroinflammation inhibition activity and an extraction method thereof. The withanolide compound with novel structure is extracted and separated from stramonium leaves, and the structural formula of the withanolide compound is shown as formula I. The activity research shows that the compound has the effect of inhibiting the release of inflammatory factor NO by nerve cells, has excellent inhibitory activity on inflammatory BV2 microglial cells induced by LPS, and can be used for preventing or treating neurodegenerative diseases such as Alzheimer disease, parkinson disease and the like.

Inventors

  • PAN JUAN
  • WANG JIESHU
  • WANG SIYI
  • XUE SIQI

Assignees

  • 黑龙江中医药大学

Dates

Publication Date
20260512
Application Date
20240122

Claims (7)

  1. 1. Compounds of formula I or a pharmaceutically acceptable salt: 。
  2. 2. a process for preparing the compound of formula I of claim 1, comprising the steps of 1) cold-leaching the leaves of Datura stramonium with methanol, recovering the solvent to obtain an extract; 2) The method comprises the following steps: a) Adding the dissolved extract into macroporous adsorption resin, eluting with water, 30% ethanol and 95% ethanol in sequence, collecting each eluent, and concentrating under reduced pressure to obtain water eluting component, 30% ethanol eluting component and 95% ethanol eluting component; b) Separating the 95% ethanol elution component by silica gel column chromatography, and performing gradient elution by taking dichloromethane-methanol as a mobile phase to obtain 8 fractions, wherein Fr.A-Fr. H; c) Separating Fr. G fraction by ODS column chromatography, and eluting with methanol-water gradient to obtain 14 fraction segments, fr. G1-Fr. G14; d) Separating Fr. G8 fraction by vacuum silica gel column chromatography, and performing gradient elution with dichloromethane-methanol to obtain 5 fraction sections, fr. G8-1-Fr.G8-5; e) The Fr. G8-3 fraction is eluted by preparative HPLC with methanol-water as eluent to obtain the compound shown in the formula I.
  3. 3. The method according to claim 2, wherein in step 1), the weight ratio of the stramonium leaf to the methanol is 1:8-12; The cold leaching extraction is carried out for 2-6 times; the extraction time is 1-7 d/time.
  4. 4. The method of claim 2, wherein in step 2), the macroporous adsorbent resin is selected from the group consisting of HPD-BJQH, HP-20, and AB-8.
  5. 5. The method of claim 2, wherein, A) The flow rate of the eluent is 1 BV.h -1 ; The water usage was 2 BV, the 30% ethanol usage was 2 BV, and the 95% ethanol usage was 4 BV; b) The elution gradient of the mobile phase dichloromethane-methanol is that the mixed solution with the volume ratio of dichloromethane to methanol of 1:0, 50:1, 20:1, 10:1, 5:1 and 0:1 is respectively eluted by 3 times, 5 times, 4 times, 3 times, 4 times and 2 times of column volume; c) The elution gradient of the mobile phase methanol-water is that the mixed solution with the volume ratio of methanol to water of 30:70, 40:60, 50:50, 60:40, 70:30, 80:20 and 1:0 is respectively eluted by 2 times, 4 times, 5 times, 3 times, 2 times and 2 times of column volume; d) The elution gradient of the mobile phase dichloromethane-methanol is that the mixed solution with the volume ratio of dichloromethane to methanol of 20:1, 15:1, 10:1 and 0:1 is respectively eluted by 2 times, 4 times, 3 times and 2 times of column volumes; e) The volume ratio of methanol to water is 65:35.
  6. 6. Use of a compound of formula I according to claim 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention and/or treatment of neuroinflammation or a neurodegenerative disease selected from alzheimer's disease or parkinson's disease by inhibiting microglial inflammatory activity.
  7. 7. A medicament for preventing and/or treating neuroinflammation or neurodegenerative disease by inhibiting microglial inflammatory activity, comprising a compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1 and a pharmaceutically acceptable carrier.

Description

Compound with neuroinflammation inhibition activity and extraction method thereof Technical Field The invention relates to the field of medicines, in particular to a compound with neuroinflammation inhibition activity and an extraction method thereof, and more particularly relates to a compound with neuroinflammation inhibition activity extracted and separated from stramonium leaves. Background The stramonium leaf is used as a dried leaf of stramonium (Datura stramonium L.) belonging to stramonium genus of Solanaceae family, has effects of relieving cough and pain, and is commonly used for treating rheumatism and rheumatoid arthritis, asthma, gastrointestinal cramp, migraine, etc., and contains a large amount of withanolides, but the research on stramonium leaf is not enough, only a small amount of withanolides and other compounds are reported in chemical aspects except alkaloids, so that further extraction, separation and identification of active ingredients in stramonium leaf are urgently needed to further study the medicinal activity of the stramonium leaf and provide more medicinal ingredients with biological activity. Disclosure of Invention In one aspect, the invention provides a compound of formula I: the compound shown in the formula I is prepared by a method comprising the following steps: 1) Cold soaking folium Daturae in methanol, and recovering solvent to obtain extract; 2) Dissolving the extract with water, and separating by column chromatography such as macroporous adsorbent resin, silica gel, ODS, gel, etc. and preparative high performance liquid chromatography to obtain withanolides compound. In the above method step 1), the weight ratio of the stramonium leaf to the methanol is 1:1-12, preferably 1:8-12, more preferably 1:10; the cold leaching may be performed a number of times, such as 2-6 times, preferably 2-4 times, more preferably 3 times; the extraction time is 1-7 d/time, preferably 2-4 d/time, more preferably 3 d/time; In step 2), the macroporous adsorbent resin is selected from HPD-BJQH, HP-20, AB-8, preferably HPD-BJQH; The operation of step 2) comprises: a) Adding the dissolved extract into macroporous adsorption resin, eluting with water, 30% ethanol (volume ratio concentration) and 95% ethanol (volume ratio concentration) in sequence, collecting each group of eluates, and concentrating under reduced pressure to obtain water eluting component, 30% ethanol eluting component and 95% ethanol eluting component; b) Separating the 95% ethanol elution component by silica gel column chromatography, and performing gradient elution (1:0-0:1, v/v) by using dichloromethane-methanol as mobile phase to obtain 8 fractions, fr.A-Fr.H; c) Separating Fr.G fraction by ODS column chromatography, eluting with methanol-water gradient (30:70-80:20, 1:0, v/v) to obtain 14 fraction segments, fr.G1-Fr.G14; d) Separating the Fr.G8 fraction by a reduced pressure silica gel column chromatography, eluting with methylene dichloride-methanol to obtain 5 fraction sections, wherein Fr.G8-1-Fr.G8-5; e) The Fr.G8-3 fraction is eluted by preparative HPLC with methanol-water as eluent to obtain the compound shown in the formula I. Wherein, in a), the flow rate of the eluent is 1 BV.h -1; The water consumption is 2BV, the ethanol consumption of 30% is 2BV, and the ethanol consumption of 95% is 4BV; b) Wherein the elution gradient of the mobile phase dichloromethane-methanol (1:0-0:1) is that the mixed solution of the dichloromethane and the methanol with the volume ratio of 1:0, 50:1, 20:1, 10:1, 5:1 and 0:1 is respectively eluted for 3 times, 5 times, 4 times, 3 times, 4 times and 2 times of column volumes; c) The elution gradient of mobile phase methanol-water (30:70-80:20, 1:0) is that the mixed solution of methanol and water with the volume ratio of 30:70, 40:60, 50:50, 60:40, 70:30, 80:20 and 1:0 is respectively eluted by 2 times, 4 times, 5 times, 3 times, 2 times and 2 times of column volume; d) The elution gradient of the mobile phase dichloromethane-methanol (20:1-0:1) is that the mixed solution with the volume ratio of dichloromethane to methanol of 20:1, 15:1, 10:1 and 0:1 is respectively eluted for 2 times, 4 times, 3 times and 2 times of column volume; e) The volume ratio of methanol to water is 65:35. In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. In still another aspect, the invention provides an application of a compound shown in formula I or a pharmaceutically acceptable salt thereof in preparing a medicament for preventing and/or treating neuroinflammation and neurodegenerative diseases. In such applications, the neurodegenerative disease includes Alzheimer's disease, parkinson's disease. The activity research shows that the compound has the effect of inhibiting the release of inflammatory factor NO by nerve cells, has excellent inhibitory activit