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CN-121974877-A - Diterpenoid compound for inhibiting NLRP3 inflammatory corpuscles and application thereof

CN121974877ACN 121974877 ACN121974877 ACN 121974877ACN-121974877-A

Abstract

The invention belongs to the field of medicines, and particularly relates to a diterpenoid compound for inhibiting NLRP3 inflammatory corpuscles and application thereof. The novel compound separated from callicarpa nudiflora is identified as diterpenoid. Cell experiments show that the compound can obviously inhibit the expression of human monocyte system THP-1 source macrophage NLRP3 inflammatory corpuscle induced by LPS, and has obvious inhibition effect on the expression of IL-1 beta and IL-18. The results show that the compound has obvious anti-inflammatory activity, provides a candidate compound for the research and development of anti-inflammatory drugs, and has important potential application value in the treatment of skin wound inflammatory diseases such as skin wound healing, diseases accompanied by inflammation or pathological processes.

Inventors

  • DONG LIN
  • YAN JINYE
  • LUO XUEMEI
  • WANG YONG
  • HE XIAOWEN

Assignees

  • 海南医科大学(海南省医学科学院)

Dates

Publication Date
20260505
Application Date
20260127

Claims (10)

  1. 1. The diterpene compound is characterized by having a structural formula as shown in formula I: 。
  2. 2. The method for extracting diterpene compound as claimed in claim 1, comprising the steps of: (1) Heating and reflux-extracting dry leaves of Callicarpa nudiflora with 95% ethanol, recovering ethanol, dissolving in water, and extracting with dichloromethane to obtain dichloromethane extract; (2) Separating the dichloromethane extract by silica gel column chromatography, and performing gradient elution by using petroleum ether and ethyl acetate mixed solution to obtain eluent; (3) Separating the fraction obtained by the volume ratio of petroleum ether to ethyl acetate of 100:10 by silica gel column chromatography, gradient eluting with mixed solution of petroleum ether and ethyl acetate, and separating and purifying the fraction eluted by the volume ratio of petroleum ether to ethyl acetate of 10:1 by semi-preparative high performance liquid chromatography to obtain the compound of formula I.
  3. 3. The extraction method of claim 2, wherein in the step (2) and the step (3), silicA gel is 200-300 meshes, in the step (2), the volume ratio of the petroleum ether and ethyl acetate mixed solution is 100:0-0:100, in the step (3), the volume ratio of the petroleum ether and ethyl acetate mixed solution is 40:1-0:100, and the semi-preparative high performance liquid chromatography column is YMC-pack ODS-A column, the column length is 250 mm, the inner diameter is 10 mm, the particle size is 5 μm, and the micropore size is 12 nm.
  4. 4. The method according to claim 3, wherein the mixed solution of petroleum ether and ethyl acetate in the step (2) has a volume ratio of 100:0, 100:1, 100:2, 100:4, 100:5, 100:8, 100:10, 100:12, 100:20, 100:50 and 0:100 in this order, the mixed solution of petroleum ether and ethyl acetate in the step (3) has a volume ratio of 40:1, 30:1, 25:1, 20:1, 15:1, 12:1, 10:1, 8:1, 5:1, 1:1 and 0:1 in this order, and the eluent in the step (3) has methanol-0.2% formic acid=73:27 in the semi-preparative high performance liquid chromatography separation process at a flow rate of 2mL/min, and the compound of formula I has a retention time of 58 minutes.
  5. 5. Use of a diterpenoid compound as defined in claim 1 in the manufacture of an anti-inflammatory medicament.
  6. 6. Use of a diterpenoid compound as defined in claim 1 in the manufacture of a medicament for promoting wound healing.
  7. 7. Use of a diterpenoid compound according to claim 1 for the preparation of a medicament for reducing the expression of NLRP3 inflammasome, IL-1 beta and/or IL-18 in cells.
  8. 8. The use according to claim 7, wherein the cell is human monocyte-derived THP-1.
  9. 9. The use according to claim 7 or 8, wherein the cells are LPS-induced human monocyte lineage THP-1 derived macrophages.
  10. 10. The use according to claim 7 or 8, wherein the diterpenoid compound is present in a concentration of 6 to 14 μm.

Description

Diterpenoid compound for inhibiting NLRP3 inflammatory corpuscles and application thereof Technical Field The invention belongs to the field of medicines, and particularly relates to a diterpenoid compound for inhibiting NLRP3 inflammatory corpuscles and application thereof. Background NLRP3 (NOD, LRR and PYD Domain protein 3) belongs to the family of nucleotide binding oligomerization domain (NOD) -like receptors (NLR), which is one of the most deeply studied inflammatory small body core components at present. NLRP3 inflammatory corpuscles are key components of the innate immune system, acting as a pro-inflammatory factor secretion platform, playing a central regulatory role in wound healing. After the wound tissue cells activate NLRP3 through Toll-like receptors and mitochondria, NF- κB pathway can be activated, and the expression of inflammatory small related proteins (such as IL-1 beta and IL-18) can be further up-regulated to prevent healing. Activation also triggers the aggregation of the inflammatory small body sensor protein with the adaptor protein, recruiting and activating Caspase-1, promoting IL-1β/IL-18 maturation, and triggering pro-inflammatory cell death. By regulating and controlling NLRP3 inflammatory small body passage, it can inhibit macrophage infiltration, promote angiogenesis, reduce IL-1 beta secretion, further improve cell hypoxia response, regulate cell proliferation and differentiation, and accelerate wound repair. Thus, inhibition of this pathway is an important strategy for treating wound healing. Chronic inflammation is closely related to arthritis, neurodegenerative diseases and the like, long-term uncontrolled control can induce DNA damage and even canceration, and regulation of inflammatory response is a key for disease treatment. The NLRP3 inflammatory corpuscles are taken as important targets for drug development, and the inhibitors thereof have potential value for preventing and treating NLRP3 inflammatory corpuscles related diseases. The research on natural medicine resources to develop high-efficiency inhibitors has great significance. The beautyberry (CALLICARPA NUDIFLORA hook. Et Arn.) is a plant of beautyberry genus (CALLICARPA L.) of Verbenaceae family, has antibacterial, hemostatic, antiinflammatory, toxic materials removing, blood stasis dispelling, repercussive, and other effects, and can be used for treating suppurative inflammation, acute infectious hepatitis, hemorrhage of respiratory tract and digestive tract, traumatic hemorrhage, etc., and various acute and chronic inflammations. The beautyberry leaf has the advantages of good anti-infection effect in burn treatment, strong convergence, capability of controlling the extravasation of wound body fluid, promotion of the growth of epithelium, acceleration of the healing of wound surface and no obvious toxicity to local and whole body. Therefore, the active ingredients for inhibiting NLRP3 inflammatory corpuscles are found from the callicarpa nudiflora, so that scientific basis is provided for revealing the anti-inflammatory substance basis and the action mechanism of the callicarpa nudiflora, and a research basis is provided for researching the precursor compounds for inhibiting NLRP3 inflammatory corpuscles of the callicarpa nudiflora. Disclosure of Invention The technical scheme of the invention is realized as follows: a diterpene compound has a structural formula shown in formula I: 。 The extraction method of the diterpenoid compound comprises the following steps: (1) Extracting dry leaves of Callicarpa nudiflora with 95% ethanol under reflux, recovering ethanol, dissolving in water, and extracting with dichloromethane to obtain dichloromethane extract; (2) Separating the dichloromethane extract by silica gel column chromatography, and performing gradient elution by using petroleum ether and ethyl acetate mixed solution to obtain eluent; (3) Separating the fraction obtained by the volume ratio of petroleum ether to ethyl acetate of 100:10 by silica gel column chromatography, gradient eluting with mixed solution of petroleum ether and ethyl acetate, and separating and purifying the fraction eluted by the volume ratio of petroleum ether to ethyl acetate of 10:1 by semi-preparative high performance liquid chromatography to obtain the compound of formula I. Further, in the step (2) and the step (3), silica gel used by a silica gel column is 200-300 meshes, the volume ratio of petroleum ether and ethyl acetate mixed solution in the step (2) is 100:0-0:100, and the volume ratio of petroleum ether and ethyl acetate mixed solution in the step (3) is 40:1-0:100. Further, in the step (2), the volume ratio of the petroleum ether and ethyl acetate mixed solution is sequentially 100:0, 100:1, 100:2, 100:4, 100:5, 100:8, 100:10, 100:12, 100:20, 100:50 and 0:100, the volume ratio of the petroleum ether and ethyl acetate mixed solution in the step (3) is sequentially 40:1, 30:1, 25:1, 20:1, 15:1, 12:1, 10:1, 8:1, 5:1, 1:1 a