Cadinane sesquiterpene compound, and preparation method and application thereof
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A technology of sesquiterpene, juniperane, applied in the field of phytochemistry
Active Publication Date: 2017-06-13
HAINAN NORMAL UNIV
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There are few reports on the juniperane-type sesquiterpenoids isolated from Melaleuca japonica, and there is no report on the application of the juniperane-type sesquiterpenoids in inhibiting the proliferation of rheumatoid arthritis synoviocytes
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Embodiment 1
[0031] The preparation method of juniperane type sesquiterpene compound, comprises the following steps:
[0032] (1) 30~95% v / v of the dried stem 10Kg of Melaleuca japonica is conventionally used, preferably 75% v / v ethanol leaching 3 times for each 7 days to obtain the extract, and the extract is heated at a temperature of 30 ~70°C and pressure of -0.06 to -0.15MPa, preferably 45°C and pressure of -0.095MPa, concentrated under reduced pressure to form a paste to obtain 1200g of guacamole extract.
[0033] (2) Dilute the extract with distilled water (3L) to form a suspension, then extract it with petroleum ether (3L×3 times) and ethyl acetate (3L×3 times) successively, and extract the ethyl acetate extract at a temperature of 30-70 ℃, pressure under the condition of -0.06~-0.15MPa, preferably 45 ℃, pressure under the condition of -0.095MPa and concentrate under reduced pressure to obtain extract (250g).
[0034] Carry out silica gel column chromatography, use petroleum ether-...
Embodiment 2
[0043] Pharmacological Activity Experiment
[0044] Experimental Materials
[0045] Cells: synoviocytes (primary rat synoviocytes).
[0046] Cell culture medium: primary rat synoviocyte culture medium.
[0047] Reagent: Thiazolium blue (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazoliumbromide, MTT, produced by Sigma). Lactate dehydrogenase (lactate dehydrogenase, LDH) kit (purchased from Beyontian Company). Methotrexate (methotrexate, MTX, Shanghai Xinyi Pharmaceutical Co., Ltd., batch number 20140307).
[0050] Determination of the Inhibitory Effect of Drugs on Synoviocytes by MTT Method
[0051] Synoviocytes in the logarithmic phase were inoculated into 96-well culture plates (1×10 4 cells / well), which were set as blank group (no drug given), methotrexate group (1 μg / mL methotrexate given), drug treatment group (1, 10, 50, 100 μg / mL drug ...
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Abstract
The invneiton provides a cadinane sesquiterpene compound, and a preparation method and application thereof. The preparation method comprises the following steps: (1) preparation of fissistigma oldhamii extract: carrying out cold leaching or hot extracting on dried stem of the fissistigma oldhamii by using ethanol of which the volume concentration is 30 to 95 percent, thus obtaining an extracting solution; carrying out vacuum concentration, thus obtaining paste, i.e., the fissistigma oldhamii extract; (2) separation and purification: diluting the fissistigma oldhamii extract by using water to prepare a suspension solution, sequentially extracting by using petroleum ether and ethyl acetate, forming extractum by carrying out vacuum concentration on an ethyl acetate extracting solution, and carrying out column chromatography, thin layer chromatography, molecular sieve chromatographic separation and high-performance liquid-phase separation, thus obtaining the cadinane sesquiterpene compound. The invention also discloses the preparation method of the cadinane sesquiterpene compound and the application of the cadinane sesquiterpene compound in preparing medicine for resisting rheumatoid arthritis. According to the cadinane sesquiterpene compound disclosed by the invention, the activity of the cadinane sesquiterpene compound in inhibiting synovial cell proliferation of the rheumatoid arthritis is firstly discovered, and the cadinane sesquiterpene compound can be used for preparing the medicine for resisting the rheumatoid arthritis.
Description
technical field [0001] The invention relates to the technical field of phytochemistry, in particular to a juniperane-type sesquiterpene compound and its preparation method and application. Background technique [0002] Fissistigma oldhamii (Hemsl.) Merr., also known as Fissistigma oldhamii (Hemsl.) Merr., is a plant of the genus Fissistigma oldhamii (Annonaceae) in the family Annonaceae. The plant can be used as medicine, the stems and leaves can cure fractures and edema, and the whole plant can treat rheumatic bone pain, numbness of hands and feet, etc. There are few reports on the juniperane-type sesquiterpenoids isolated from Melaleuca japonica, and there is no report on the application of the juniperane-type sesquiterpenoids in inhibiting the proliferation of rheumatoid arthritis synoviocytes. Contents of the invention [0003] In view of this, the object of the present invention is to provide a kind of juniperane type sesquiterpene compound that extracts and separate...
Claims
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