CN-121974873-A - Diterpenoid compound and preparation method and application thereof

Abstract

The invention provides a diterpenoid compound, a preparation method and application thereof, and belongs to the technical field of biological medicines. The invention utilizes Bipolaris sp of Bipolaris to ferment rice to obtain rice ferment, the rice ferment is extracted by ethanol to obtain ethanol extract, the ethanol extract is extracted by ethyl acetate to obtain ethyl acetate extract, and diterpenoid compounds (compound 1-compound 7) are obtained by separating the ethyl acetate extract. The compounds 1-6 provided by the invention are enantiomer-kaurane diterpenes which form a 5/6/6/5 ring system after C-2 is degraded, and the compounds 7 are enantiomer-kaurane diterpenes which form epoxy at C-2 and C-3. The compound provided by the invention can inhibit the P-gp transport function and reduce the excretion of paclitaxel, thereby obviously improving the therapeutic effect of paclitaxel.

Inventors

• ZHANG YONGHUI
• ZHU HUCHENG
• CHEN CHUNMEI
• SHEN YONG
• LI QIN
• SUN WEIGUANG
• Ding Nanjin

Assignees

• 华中科技大学

Dates

Publication Date

20260505

Application Date

20260127

Claims (8)

1. 1. A diterpenoid compound is characterized by having a structure shown in a compound 1-a compound 7: 。
2. 2. a process for the preparation of diterpenoids according to claim 1, comprising the steps of: Fermenting rice with Bipolaris sp to obtain rice ferment; extracting the rice ferment with ethanol to obtain ethanol extract; extracting the ethanol extract with ethyl acetate to obtain an ethyl acetate extract; And separating the ethyl acetate extract to obtain the diterpenoid compound.
3. 3. The preparation method according to claim 2, characterized in that the fermentation is in particular: after mixing rice with water, the Bipolaris sp. Strain was inoculated, followed by fermentation at 28 ℃ for 30 days.
4. 4. The method of preparation according to claim 2, wherein the separation comprises the steps of: (1) Subjecting the ethyl acetate extract to normal phase silica gel column chromatography, gradient eluting with petroleum ether, ethyl acetate and methanol mixed solvent at volume ratio of 10:1:0-5:5:2, and detecting and combining similar parts by thin layer chromatography to obtain Fr.1-Fr.5 total 5 components; (2) Subjecting the Fr.3 to reverse phase C 18 column chromatography, performing gradient elution with mixed solvent of methanol and water, wherein methanol accounts for 30% -100% of the mixed solvent to obtain 11 components, subjecting the Fr.3.8 to normal phase silica gel column chromatography, performing gradient elution with mixed solvent of petroleum ether and ethyl acetate with volume ratio of 15:1-0:1 to obtain 13 components, subjecting the Fr.3.8.11 to normal phase silica gel column chromatography, and performing gradient elution with mixed solvent of petroleum ether and ethyl acetate with volume ratio of 10:1-0:1 to obtain 8 components, wherein Fr.3.8.11.2 is subjected to reverse phase high performance liquid chromatography to obtain compound 1; (3) Purifying Fr.3.8.11.3 by reverse phase high performance liquid chromatography to obtain a compound 2; (4) Subjecting Fr.3.7 to normal phase silica gel column chromatography, and gradient eluting with mixed solvent of petroleum ether and ethyl acetate at volume ratio of 15:1-0:1 to obtain 13 components Fr.3.7.1-Fr.3.7.13, and separating from Fr.3.7.1 by reversed phase high performance liquid chromatography to obtain compound 4 and compound 6; (5) Purifying Fr.3.8.5 by reverse phase high performance liquid chromatography to obtain a compound 7; (6) The Fr.4 is subjected to reverse phase column chromatography, gradient elution is carried out by using a mixed solvent of methanol and water, wherein the volume ratio of the methanol to the mixed solvent is 30% -100%, the Fr.4.1-Fr.4.19 total 19 components are obtained through separation, the Fr.4.17 is subjected to normal phase silica gel column chromatography, the Fr.4.17.1-Fr.4.17.8 total 8 components are obtained through gradient elution by using a mixed solvent of dichloromethane and methanol with the volume ratio of 100:1-0:1, the Fr.4.17.7 is subjected to normal phase silica gel column chromatography, gradient elution is carried out by using a mixed solvent of petroleum ether and ethyl acetate with the volume ratio of 10:1-0:1, the Fr.4.17.1-Fr.17.7.5 total 5 components are obtained, and the Fr.4.17.7.3 is separated by reverse phase high-performance liquid chromatography, so that the compound 3 and the compound 5 are obtained.
5. 5. A medicament for reversing the resistance of colon cancer cells to paclitaxel comprising the diterpenoid compound of claim 1.
6. 6. Use of a diterpenoid compound of claim 1 in the manufacture of a medicament for reversing the resistance of colon cancer cells to paclitaxel.
7. 7. A P-gp inhibitor comprising a diterpenoid compound according to claim 1.
8. 8. Use of a diterpenoid compound according to claim 1 for the preparation of a P-gp inhibitor drug.

Description

Diterpenoid compound and preparation method and application thereof Technical Field The invention relates to the technical field of biological medicine, in particular to a diterpenoid compound and a preparation method and application thereof. Background Malignant tumors seriously threaten human health, and the current main treatment means comprise surgery, radiotherapy, chemotherapy, targeted therapy, immunotherapy and the like. Although the curative effect of surgical excision is clear, the method is only suitable for 10% -20% of patients, the postoperative patients are easy to relapse, the effect of the method on the patients in the late stage of metastasis is limited, the sensitive tumor can be controlled locally by radiotherapy, but the focus is difficult to eradicate, the body functions can be damaged, and the targeting and immunotherapy have the limitations of narrow applicable crowd, long treatment course, high cost and the like. Thus, chemotherapy is still one of the important ways of clinically treating malignant tumors. However, various chemotherapeutic drugs including vinblastine, aclarubicin, etc. are extremely susceptible to drug resistance, and even the first-line drug paclitaxel, drug resistance occurs in various tumors. The problem of tumor resistance has become a major cause of treatment failure and patient death. Tumor resistance is classified into original drug resistance and multi-drug resistance. The original drug resistance only refers to drug resistance to an induced drug, and the multi-drug resistance (MDR) is generated by long-term contact of tumor cells with a certain chemotherapeutic drug, can not only resist the chemotherapeutic drug, but also generate cross drug resistance to a plurality of chemotherapeutic drugs with different structures and functions, is the most important defense mechanism of the tumor cells from being attacked by the chemotherapeutic drug, and is also one of the main reasons for causing chemotherapy failure. Therefore, drug studies aimed at reversing tumor resistance have become an important point in anti-tumor studies. Over-expression of P-glycoprotein (P-gp) is a key mechanism of tumor multi-drug resistance, which leads to treatment failure mainly by actively excreting chemotherapeutic drugs, lowering intracellular drug concentration. To overcome this resistance, third generation P-gp inhibitors have been developed in tandem, first generation such as verapamil, cyclosporin A, second generation such as verapamil, third generation such as zosuquidar, tariquidar, etc. However, these inhibitors generally have problems of low selectivity, undesirable pharmacokinetics or obvious toxic and side effects, and most of them have not been successfully applied to clinic. Therefore, the development of novel P-gp inhibitors that are highly potent and low in toxicity remains an urgent need to overcome multi-drug resistance. Disclosure of Invention In view of the above, the present invention aims to provide a diterpenoid compound, and a preparation method and application thereof. In order to achieve the above object, the present invention provides the following technical solutions: According to one of the technical schemes, the diterpenoid compound has the structure shown as the compounds 1-7: 。 the second technical scheme of the invention is that the preparation method of the diterpenoid compound comprises the following steps: Fermenting rice with bipolaris fungus to obtain rice ferment; extracting the rice ferment with ethanol to obtain ethanol extract; extracting the ethanol extract with ethyl acetate to obtain an ethyl acetate extract; And separating the ethyl acetate extract to obtain the diterpenoid compound. In a third technical scheme of the invention, a medicine for reversing the drug resistance of colon cancer cells to paclitaxel comprises the diterpenoid compound. The fourth technical scheme of the invention is the application of the diterpenoid compound in preparing medicines for reversing the drug resistance of colon cancer cells to taxol. The fifth technical scheme of the invention is that the P-gp inhibitor comprises the diterpenoid compound. The sixth technical scheme of the invention is the application of the diterpenoid compound in preparing a P-gp inhibitor drug. The invention discloses the following technical effects: The compounds 1-6 provided by the invention are enantiomer-kaurane diterpenes which form a 5/6/6/5 ring system after C-2 is degraded, and the compounds 7 are enantiomer-kaurane diterpenes which form epoxy at C-2 and C-3. The compound provided by the invention can inhibit the P-gp transport function and reduce the discharge of paclitaxel, thereby obviously improving the therapeutic effect of paclitaxel and providing an important thought for searching a new lead compound to overcome the drug resistance of paclitaxel. Drawings In order to more clearly illustrate the embodiments of the present invention or the technical solutions of the prior