阅读说明：本技术 降碳贝壳杉烷型二萜及其制备方法和在制备抗肿瘤药物中的用途 (Carbon-reduced kaurane diterpene, preparation method thereof and application thereof in preparation of antitumor drugs ) 是由 胡金锋 李�浩 熊娟 于 2019-02-12 设计创作，主要内容包括：本发明属医药技术领域,涉及降碳贝壳杉烷型二萜,尤其是C-18位降碳的贝壳杉烷型(18-nor-kaurane-type)二萜类化合物其制备方法和在制备抗肿瘤药物中的用途。本发明从穗花杉(Amentotaxus argotaenia)的枝叶中提取分离得到的C-18位降碳且C-4与C-19位形成三元氧环的贝壳杉烷型二萜类化合物amentotanol A和amentotanol B,经体外生物活性测试表明,可显著抑制多种人肿瘤细胞的增殖,可制备治疗临床常见多发肿瘤,如宫颈癌、卵巢癌、乳腺癌、肺癌、肝癌或结肠癌的药物,和抗肿瘤药物的先导化合物。<Image he="438" wi="582" file="DDA0001968300080000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>(The invention belongs to the technical field of medicines, and relates to a carbon-reducing kaurane diterpenoid, in particular to a C-18 carbon-reducing kaurane (18-nor-kaurane-type) diterpenoid compound, a preparation method thereof and application thereof in preparing antitumor drugs. The kaurane diterpenoid compounds, namely, the amentotanol A and the amentotanol B, which are extracted and separated from branches and leaves of amentoflavone (Amentotaxus argotaenia) and have C-18 position carbon reduction and C-4 and C-19 positions form ternary oxygen rings are shown by in vitro biological activity tests to be capable of obviously inhibiting the proliferation of various human tumor cells and being capable of preparing and treating clinical diseasesCommon multiple tumors, such as cervical cancer, ovarian cancer, breast cancer, lung cancer, liver cancer or colon cancer, and lead compounds of antitumor drugs.)

1. The carbon-reduced kaurane diterpenoid compound amentotanol A or amentotanol B has the following structural formula, and is characterized in that the C-18 position of the compound is carbon-reduced, and the C-4 position and the C-19 position form a unique ternary oxygen ring.

2. The carbon-reduced kaurane-type diterpenoid compound of claim 1, which is prepared by the following method:

drying branches and leaves of amentoflavone, crushing, extracting with 90% methanol, concentrating the extracting solution, adding water for suspension, extracting with petroleum ether, ethyl acetate and n-butyl alcohol respectively, concentrating the ethyl acetate extracting solution under reduced pressure to obtain an extract, and separating and preparing the extract by silica gel column chromatography, MCI microporous resin column chromatography, Sephadex LH-20 gel column chromatography and reversed-phase semi-preparative HPLC chromatography to obtain compounds of amentotanol A and amentotanol B.

3. Use of the carbon-reduced kaurane diterpenoid compound amentotanol A or amentotanol B as claimed in claim 1 in the preparation of a medicament for treating tumors.

4. The use of claim 3, wherein the tumor is cervical cancer, ovarian cancer, breast cancer, lung cancer, liver cancer, or colon cancer.

5. Use according to claims 3 and 4, characterized in that the compound ametotanol A or ametotanol B is used alone or in combination.

6. The use according to claims 3 and 4, characterized in that the compound ametotanol A or ametotanol B, respectively, is combined with excipients to make tablets, capsules, granules or injections.

Technical Field

The invention belongs to the technical field of medicines, and relates to a carbon-reducing kaurane diterpenoid, in particular to a C-18 carbon-reducing kaurane (18-nor-kaurane-type) diterpenoid compound, a preparation method thereof and application thereof in preparing antitumor drugs. The kaurane diterpenoid compounds, namely, the amentotanol A and the amentotanol B, which are extracted and separated from branches and leaves of amentoflavone (Amentotaxus argotaenia) and have C-18 position carbon reduction and C-4 and C-19 positions form a three-membered oxygen ring are shown by in vitro biological activity tests to be capable of obviously inhibiting the proliferation of various human tumor cells and being capable of preparing medicines for treating common clinical multiple cancers and lead compounds of antitumor medicines.

Background

Disclosure of Invention

The invention aims to provide a novel carbon-reduced kaurane-type diterpenoid, in particular to a C-18 carbon-reduced kaurane-type (18-nor-kaurane-type) diterpenoid compound, a preparation method thereof and application thereof in preparing antitumor drugs based on the current situation of the prior art. The invention extracts and separates new carbon-reduced kaurane diterpenoid compounds from amentoflavone, and pharmacological activity tests show that the compounds have obvious activity of inhibiting tumor cell proliferation, can be used for preparing medicaments for treating tumors and can be used as lead compounds for potential antitumor medicament research.

The invention extracts and separates the branch and leaf of the plant amentoflavone [ Amentotaxus argotaenia (Hance) Pilger ] of the genus amentoflavone of the family Taxaceae (Taxaceae) to obtain the kaurane diterpene amentotanol A and the amentotanol B which have carbon reduction at the C-18 position and form a ternary oxygen ring at the C-4 position and the C-19 position;

the kaurane diterpenoid compound has the following chemical structural formula:

another object of the present invention is to provide a process for the preparation of the compound.

The compound is prepared from amentoflavone branches and leaves by a conventional extraction and separation method related in the field, and comprises the following steps: crushing dried branches and leaves of amentoflavone, soaking and extracting the branches and leaves of the amentoflavone with methanol at room temperature, decompressing and concentrating an extracting solution, combining the extracting solution after multiple times of extraction and concentration to obtain an extract, then dispersing the extract with water, sequentially extracting the extract with petroleum ether, ethyl acetate and n-butyl alcohol solvents, and separating the ethyl acetate extract after decompressing and concentrating by silica gel, microporous resin, gel and semi-prepared high-efficiency liquid phase to obtain compounds of amentotanol A and amentotanol B.

The compound of the invention can be obtained by separating and purifying from plants; can also be obtained by chemical synthesis methods well known to those skilled in the art.

The compound can obviously inhibit the proliferation of 6 tumor cells such as human cervical cancer cell HeLa, human lung cancer cell A549, human liver cancer cell Huh-7, human breast cancer cell MDA-MB-231, human ovarian cancer cell SKOV3 and human colon cancer cell HCT-116, and the result shows that the compound can be used for preparing an anti-tumor medicament or used as a potential anti-tumor medicament lead compound.

The anti-tumor medicament is a medicament for treating cervical cancer, ovarian cancer, breast cancer, lung cancer, liver cancer or colon cancer.

The compound can be used alone or in combination, or can be combined with pharmaceutically suitable carriers or excipients to be prepared into oral or non-oral dosage forms according to a conventional method.

The invention has the following advantages: the target compound is a carbon-reduced kaurene diterpenoid compound with a novel structure, wherein carbon is reduced at a C-18 position, and a unique ternary oxygen ring is formed at a C-4 position and a C-19 position; the compound has obvious activity of inhibiting tumor cell proliferation, and has application prospect in the development of anti-tumor drugs for the tumor diseases with high attention, such as cervical cancer, ovarian cancer, breast cancer, lung cancer, liver cancer or colon cancer.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to be limiting in any way. Any variations that may be made in the practice of the invention by those skilled in the art in light of the teachings herein will fall within the scope of the appended claims.

In the following preparation examples, the specific rotation test was carried out by a Rudolph Autopol VI polarimeter; ultraviolet and infrared are respectively measured by a Hitachi U-2900E double-beam spectrophotometer and a Thermo Scientific Nicolet Is5 Fourier transform infrared spectrometer; measuring the circular dichroism spectrum by adopting a JASCO-810 spectropolarimeter; the nuclear magnetic resonance spectrum is measured by adopting a Bruker Avance III400MHz nuclear magnetic resonance instrument and a Bruker Avance DRX-600MHz nuclear magnetic resonance instrument; chemical shifts are expressed in (ppm) and referenced to residual solvent peaks; ESI-MS adopts Agilent 1100series mass spectrometer to determine; high resolution mass spectrometry was determined using an Agilent technologies 6224TOF LC/MS mass spectrometer; semi-preparative high performance liquid chromatography apparatus: waters e2695 series, equipped with Waters 2998 PDA and Waters 2424ELSD detectors; semi-preparative chromatography column: sunfire C₁₈OBDPrep Column (5 μm,10mm × 250mm, flow rate: 3.0Ml/min), normal phase Column chromatography silica gel specification of 100-200 mesh (Nitai Muyun Hebino chemical reagent factory and Qingdao oceanic chemical plant), gel Sephadex LH-20(GEHealthcare Bio-Sciences AB, Sweden), Mitsubishi gel adsorption resin MCI gel CHP20P specification of 75-150 μm (Mitsubishi chemical Industries, Tokyo, Japan), thin layer chromatography plate (TLC) specification of GF₂₅₄0.25mm (tobacco bench, muyun-hei, conduo chemical reagent factory); all reagents are produced by Shanghai national drug group chemical reagent limited.