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CN-121990923-A - Diterpenoid compound, extraction method thereof and application thereof in preparation of anti-inflammatory drugs

CN121990923ACN 121990923 ACN121990923 ACN 121990923ACN-121990923-A

Abstract

The invention discloses diterpenoid compounds, an extraction method thereof and application thereof in preparing anti-inflammatory drugs, belongs to the field of traditional Chinese medicine extraction, and relates to diterpenoid compounds or pharmaceutically acceptable salts thereof or pharmaceutical compositions containing the diterpenoid compounds have an inhibitory effect on NO generation in RAW264.7 cells induced by LPS, so that the diterpenoid compounds can be applied to preparing anti-inflammatory drugs. The method of the invention further enriches the structural diversity of the euphorbia plant, lays a foundation for carrying out relevant biological activity test on the subsequently obtained monomer compound, provides an active lead compound for new drug development, and simultaneously provides a theoretical basis for deep research and development of the euphorbia plant.

Inventors

  • CHEN LIXIA
  • LI HUA
  • LIU YING
  • XU YANG
  • REN LINGXIAO

Assignees

  • 沈阳药科大学

Dates

Publication Date
20260508
Application Date
20260303

Claims (8)

  1. 1. Diterpenoid compound or pharmaceutically acceptable salt thereof, characterized in that the diterpenoid compound is selected from the following compounds: 。
  2. 2. a method of extracting a diterpenoid compound or a pharmaceutically acceptable salt thereof according to claim 1, comprising the steps of: (1) Taking euphorbia pekinensis whole herb as a raw material, adding an ethanol aqueous solution with the mass of 8-10 times of the raw material, carrying out reflux extraction, recovering a solvent from an extracting solution under reduced pressure, and concentrating to obtain a total extract; (2) Dispersing the total extract into water with the mass of 2-6 times of that of the total extract, sequentially extracting with petroleum ether and ethyl acetate respectively, concentrating the extract to recover the solvent to obtain petroleum ether extract concentrate, ethyl acetate extract concentrate and water respectively; (3) Separating petroleum ether extraction concentrate by polyamide column chromatography, gradient eluting with ethanol-water as eluent at volume ratio of 100:1-0:1, and collecting crude fraction with ethanol volume fraction of 80% -100%; (4) Separating the crude fraction by silica gel column chromatography, gradient eluting with petroleum ether-ethyl acetate as eluent at volume ratio of 100:1-0:1, and collecting fraction E4 at volume ratio of 10:1-5:1; (5) Concentrating fraction E4, separating the concentrated solution by MCI column chromatography, gradient eluting with methanol-water as eluent with volume ratio of 100:1-0:1, and collecting fraction E41 with methanol volume fraction of 90% -100%; (6) Concentrating fraction E41, separating the concentrated solution by silica gel column chromatography, gradient eluting with petroleum ether-acetone as eluent at volume ratio of 100:1-0:1, and collecting fraction E412 at volume ratio of 20:1-8:1; (7) Concentrating fraction E412, separating the concentrated solution by silica gel column chromatography, gradient eluting with petroleum ether-acetone as eluent at volume ratio of 100:1-0:1, and collecting fraction E4122 at volume ratio of 20:1-5:1; (8) Concentrating fraction E4122, separating the concentrate by ODS column chromatography, and further purifying to obtain diterpenoid compounds 1,2, 3 and 4.
  3. 3. The method for extracting diterpenoid compounds or pharmaceutically acceptable salts thereof according to claim 2, wherein in the step (1), euphorbia pekinensis is taken as a raw material, an ethanol aqueous solution with the volume concentration of 60% -95% which is 8-10 times of the raw material is added, the reflux extraction is carried out for 2-4 times, each time for 2-4 hours, the extracting solutions are obtained by combining, the solvent is recovered under reduced pressure, and the total extract is obtained after concentration.
  4. 4. The method for extracting diterpenoid compounds or pharmaceutically acceptable salts thereof according to claim 2, wherein the specific separation and purification process of fraction E4122 in step (8) is as follows: Concentrating the fraction E4122, performing ODS column chromatography separation, performing gradient elution by taking methanol-water with the volume ratio of 50:50-100:0 as an eluent, collecting the fraction with the volume ratio of methanol-water of 70:30, recording as E41222, concentrating the fraction E41222, performing gradient elution by taking petroleum ether-acetone with the volume ratio of 100:1-0:1 as the eluent, collecting the fraction with the volume ratio of petroleum ether-acetone of 20:1-8:1, recording as E412222, performing Sephadex LH-20 gel chromatography on the fraction E412222, taking methanol as an eluting solvent, performing preparative HPLC chromatography purification, and separating to obtain a compound 1, a compound 3 and a compound 4 by taking acetonitrile-water with the volume ratio of 60:40 as a mobile phase; Concentrating fraction E4122, separating by ODS column chromatography, gradient eluting with methanol-water as eluent at volume ratio of 50:50-100:0, collecting fraction with methanol-water volume ratio of 50:50, denoted as E41221, concentrating fraction E41221, subjecting to Sephadex LH-20 gel chromatography with methanol as eluting solvent, purifying by preparative HPLC chromatography, and separating with methanol-water as mobile phase at volume ratio of 65:35 to obtain compound 2.
  5. 5. A pharmaceutical composition, characterized in that, the pharmaceutical composition comprises one or more diterpenoid compounds or pharmaceutically acceptable salts thereof according to claim 1.
  6. 6. A pharmaceutical preparation comprising one or more diterpenoid compounds according to claim 1 or pharmaceutically acceptable salts thereof as an active ingredient, and pharmaceutically acceptable excipients.
  7. 7. The pharmaceutical preparation according to claim 6, wherein the administration route of the pharmaceutical preparation is oral administration or injection, and the dosage form is tablet, capsule, powder, syrup or injection.
  8. 8. Use of a diterpenoid compound according to claim 1 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 5 or a pharmaceutical formulation according to claim 6 or 7 for the preparation of an anti-inflammatory medicament.

Description

Diterpenoid compound, extraction method thereof and application thereof in preparation of anti-inflammatory drugs Technical Field The invention belongs to the field of traditional Chinese medicine extraction, and in particular relates to diterpenoid compounds separated from euphorbia pekinensis, an extraction method thereof and application thereof in preparing anti-inflammatory medicines. Background Euphorbia pekinensis (Euphornia peplus Linn.) commonly known as Phyllostachys praecox or Euphorbia pekinensis is an annual small weed characterized by its white milk. The plant is native to europe and north africa, especially in the mediterranean region, and has been widely distributed worldwide due to its rapid growth and strong invasiveness. In China, the plant has been introduced into Yunnan, guangdong and other places （Hua J, Liu Y, Xiao CJ,et al. Chemical profile and defensive function of the latex ofEuphorbia peplus[J]. Phytochemistry, 2017, 136: 56-64; Pu XX, Ran XQ, Yan Y,et al. Three new jatrophane diterpenoids fromEuphorbia peplus Linn. with activity towards autophagic flux [J]. Phytochemistry Letter, 2022, 50: 141-146）. in traditional folk medicine, and the euphorbia pekinensis is widely used for treating various diseases, including dermatopathy, diabetes, asthma, inflammation, tumor and other places （Anastasiou E, Lorentz KO, Stein GJ,et al.Prehistoric schistosomiasis parasite found in the Middle East [J]. The Lancet Infectious Diseases, 2014, 14: 553-554; Chen H, Wang H, Yang B,et al. Diterpenes inhibiting NO production fromEuphorbia helioscopia [J]. Fitoterapia, 2014, 95: 133-138.）. aiming at euphorbia pekinensis herb, and the plant is rich in diterpenoid compounds, including compounds with ingene type, jatrophane type, petiolane type (pepluane type), paraelene type (paraliane type) and ent-abine type frameworks, and shows various biological activities, such as （Ali AA, Sayed HM, Ibrahim SRM,et al.Chemical constituents, antimicrobial, analgesic, antipyretic, and anti-inflammatory activities ofEuphorbia peplusL [J]. Phytopharmacology, 2013, 4: 69-80; Chen L, Liu L, Li Y,et al.Macrocyclic Diterpenoids fromEuphorbia peplusPossessing Activity Towards Autophagic Flux [J]. International Journal of Molecular Sciences, 2025, 26: 299; Gao Y, Zhou JS, Liu HC,et al.Phonerilins A-K, cytotoxic ingenane and ingol diterpenoids fromEuphorbia neriifolia [J]. Tetrahedron, 2022, 123: 132955; Li Y, Yu ZP, Li YP,et al.Diterpenoids fromEuphorbia pepluspossessing cytotoxic and anti-inflammatory activities [J]. Bioorganic Chemistry, 145: 107194; Rizk AM, Hammouda FM, El-Missiry MM,et al.Biologically active diterpene esters fromeuphorbia peplus [J]. Phytochemistry, 24: 1605-1606; Wan LS, Chu R, Peng XR,et al.Pepluane and Paraliane Diterpenoids fromEuphorbia pepluswith Potential Anti-inflammatory Activity [J]. Journal of Natural Products, 2016, 79: 1628-1634; Wang W, Xiong L, Wu Y,et al. New lathyrane diterpenoid hybrids have anti-inflammatory activity through the NF-κB signaling pathway and autophagy [J]. Acta Materia Medica, 2022, 1: 224-243; Yan Y, Peng MY, Yang Y,et al.Highly oxygenated ent-abietane diterpenoid lactones fromEuphorbia peplusand their anti-inflammatory activity [J]. Bioorganic Chemistry, 2025, 154: 107989; Yan Y, Zhou Q, Ran X,et al.Jatrophane Diterpenoids fromEuphorbia peplusLinn. as Activators of Autophagy and Inhibitors of Tau Pathology [J]. International Journal of Molecular Sciences, 2023, 24: 1088; Yang Y, Zhou M, Wang D,et al.Jatrophane Diterpenoids fromEuphorbia peplusas Multidrug Resistance Modulators with Inhibitory Effects on the ATR-Chk-1 Pathway [J]. Journal of Natural Products, 2021, 84: 339-351）. of anticancer, anti-inflammatory, antibacterial, multi-drug resistance reversing activity and the like, in order to exert the maximum medicinal value of euphorbia pekinensis, systematic component research is carried out on euphorbia pekinensis, novel diterpenoid compounds are obtained by separation, the structures of the compounds are confirmed by means of nuclear magnetism, mass spectrum and other means, and the inhibition effect of the extracted compounds on NO production in RAW264.7 cells induced by LPS is detected. Disclosure of Invention The primary object of the present invention is to provide four diterpenoids. The second object of the present invention is to provide a method for extracting said diterpenoid compounds. It is a third object of the present invention to provide a pharmaceutical composition containing said diterpenoid compounds. A fourth object of the present invention is to provide the use of said diterpenoid compounds or of said compounds in the preparation of anti-inflammatory drugs, or of pharmaceutically acceptable salts thereof or of pharmaceutical compositions comprising said compounds. In order to achieve the above object, the technical scheme of the present invention is summarized as follows: diterpenoid compounds represented by formula 1,2, 3 or 4 isolated from Euphorbi