Application of betulinic acid derivatives in preparation of medicine for treating cardiovascular diseases
A betulinic acid and cardiovascular technology, applied in the field of medicine, can solve the problems of lack of active groups in the nucleus, few reaction sites, and less research on betulinic acid derivatives, so as to achieve good cardiomyocyte protection activity, Widely Useful Effects
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Embodiment 1
[0019] Preparation of compounds of formula formula Ⅰ- formula XVI: Example 1
[0020] The present invention is a method using a microbial conversion to betulin acid as raw material, after fermentation, extraction and separation steps, to prepare compounds of the present invention. Rhizopus (Rhizopus) are available commercially from a strain of the genus Culture Collection Center, Chinese Academy (CGMCC), potato selection medium, stored in the solid slant culture on opposite 4 ℃ refrigerator.
[0021] In Rhizopus arrhizus Rhizopus arrhizus CGMCC 3.868 as an example, the preparation of the compounds of the formula formula formula XVI Ⅰ- follows:
[0022] 1) Fermentation, transformation, and extraction
[0023] The Rhizopus arrhizus Rhizopus arrhizus CGMCC 3.868 Access two 250mL flask (with potato 100mL medium), as a seed liquid. After 1 day of shaking on the shaker, 26 ° C was oscillated for 1 day, the growth of the bacteria was in a strong period, and 1 ml of seed fluid was suckled...
Embodiment 2
[0054] Example 2: Compound I-XVI on myocardial cell protection activity damaged by hydroperoxide
[0055] (1) Experimental materials
[0056] CO 2Culture Box (Jouan IGO150); enzyme standard (Bio-Tek Elx800); fluorescence inverted microscope (Olympusix51); MTT cell proliferation and cytotoxic detection kit (Biyun Sky Biotechnology Research Institute), DMEM High Sugar Media (Gibcol BRL ), Fetal bovine serum, dimethyl sulfoxide (DMSO), trypsin (Shanghai Bioengineering Co., Ltd.), 30% hydrogen peroxide (H 2 O 2 (Tianjin Ruijin Characterization Co., Ltd.), H9C2 cells (Institute of Tumor, Chinese Academy of Medical Sciences).
[0057] (2) Experimental method
[0058] Each trial compound is determined by MTT method 2 O 2 Influence of damaged H9C2 cell activity: After digestion with pancreatic enzyme, the cellular density of cell suspension is adjusted to 5 × 10 4 2 μl of each well in 96-well culture plates, placed in 5% CO 2 , 37 ° C constant temperature CO 2 The culture tank was culture...
Embodiment 3
[0066] Example 3 Protective effect of Compound I-XVI on myocardial cytracychemia and reperfusion injury
[0067] 1) Experimental materials
[0068] CO 2 Culture Box (Jouan IGO150); enzyme standard (Bio-Tek Elx800); fluorescence inverted microscope (Olympusix51); MTT cell proliferation and cytotoxic detection kit (Biyun Sky Biotechnology Research Institute), DMEM High Sugar Media (Gibcol BRL ), Fetal bovine serum, dimethyl sulfoxide (DMSO), trypsin (Shanghai Biological Engineering Co., Ltd.), H9C2 Cells (Institute of Tumors, Chinese Academy of Medical Sciences).
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