阅读说明：本技术 源于草酸青霉的iso-Penicillixanthone A及抗顺铂耐药的应用 (iso-Penicillium xanthone A from Penicillium oxalicum and application of anti-cisplatin drug resistance ) 是由 郑秋红 陈立 刘沁颖 李欣欣 程苗苗 王思远 于 2018-12-04 设计创作，主要内容包括：本发明涉及一种源于草酸青霉的iso-Penicillixanthone A及其抗顺铂耐药方面的应用。该化合物结构特征是：<Image he="255" wi="309" file="DEST_PATH_IMAGE002.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>,与已知物青霉蒽酮A(Penicillixanthone A)互为对映体。经实验证实,所述蒽醌类化合物对顺铂耐药细胞具有较好的抑制活性。可作为制备顺铂耐药细胞增殖抑制药物或抗顺铂耐药药物用于抗肿瘤的研究。(The invention relates to iso-Penicillium xanthene A from Penicillium oxalicum and application thereof in the aspect of cisplatin resistance. The compound is characterized by comprising the following structural characteristics: in the form of an enantiomer with the known substance Penicillium anthrone A (Penicillium xanthone A). Experiments prove that the anthraquinone compound has better inhibitory activity on cisplatin-resistant cells. Can be used for preparing cisplatin-resistant cell proliferation inhibiting drugs or cisplatin-resistant drugs for antitumor research.)

1. Compound (I)The method is characterized in that: the compound is prepared by the following method: fermented Penicillium oxalicum (Penicillium oxalicum) IBPT-6, obtaining a fermentation product, and then separating and purifying the fermentation product to obtain the compound; wherein said Penicillium oxalicum (A), (B)Penicillium oxalicum) IBPT-6, which has been deposited at the China center for type culture Collection on 25.12.2013, address: wuhan, Wuhan university, the accession number is: CCTCC NO: m2013714.

2. Use of a compound according to claim 1 for the preparation of a cisplatin-resistant cell proliferation inhibitory drug.

3. The use of a compound according to claim 1 for the manufacture of a medicament against cisplatin resistance.

Technical Field

The invention relates to iso-Penicillium xanthene A derived from penicillium oxalicum and application thereof in the aspect of cisplatin resistance, belonging to the field of biological medicine.

Background

In the current clinical treatment, a plurality of tumors have better effects through primary drug treatment, but the tumors are frequently relapsed finally, the sensitivity of the antitumor drugs after the relapse is obviously reduced, and the survival quality and the survival time of patients are seriously influenced. Cisplatin (DDP) was first synthesized by the young Douling chemist Michele Payrone in 1844. However, until 1892, its structure was not known by the scientist Alfred Werner in zurich. The product can bind with DNA to cause cross-linking, thereby destroying DNA function and inhibiting cell mitosis, and is a cell non-specific medicine. The product has wide antitumor spectrum (such as various solid tumors including lung cancer, ovarian cancer, prostatic cancer, nasopharyngeal carcinoma, esophageal carcinoma, and malignant lymphoma). Cisplatin is one of the chemotherapy drugs commonly used in clinic, but it has great side effects and chemotherapy is often limited by intrinsic and acquired resistance.

Penicillium anthrone is an anthraquinone organic compound produced by secondary metabolism of fungi, and is first obtained from Penicillium anthrone A in 2002Penicillium thomiiTo date, over a decade has passed. The compounds are reported to date as 2, each of which is penam A, B and all of which are 2-4' linked. The compound has obvious inhibition on tumor cells, and is an ideal raw material for developing antitumor drugs or tumor cell proliferation inhibitors. But the research on the antitumor activity of their enantiomers is still blank.

The present inventors have studied and found that Penicillium oxalicum (Penicillium oxalicum) IBPT-6, (deposited at the chinese type culture collection on 25.12.2013, address: wuhan, Wuhan university, the accession number is: CCTCC NO: m2013714) has a good cell proliferation inhibitory activity, and the active ingredients thereof were investigated. The research finds that the penicillanic anthrone compound has the advantages ofThe activity of anti-human cisplatin-resistant cell strain is not reported at present, and the proliferation inhibition activity of the compound on human cisplatin-resistant cells is not reported, so that no related medicine is seen on the market.

Disclosure of Invention

The invention aims to provide iso-Penicillium xanthone A derived from penicillium oxalicum and application thereof in the aspect of cisplatin resistance.

In order to realize the purpose, the following technical scheme is adopted:

the iso-Penicillium xanthone A derived from Penicillium oxalicum has the effect of inhibiting the proliferation of cisplatin-resistant cells and has anti-human cisplatin-resistant activity. The structural formula is as follows:

。

the preparation method of the compound is to culture penicillium oxalicum (A)Penicillium oxalicum) IBPT-6, obtaining fermented mycelium, and separating and purifying the compound from the mycelium. The method comprises the following specific steps:

1 fermentation production

Culturing microorganism by conventional method, collecting Penicillium oxalicum (B) ((B))Penicillium oxalicum) IBPT-6 is inoculated on a flat solid culture medium, cultured in an incubator at 28 ℃ for 2-3 d, then inoculated in a fungus culture medium, statically cultured at 28 ℃ for 30 d, and filtered by a plurality of layers of gauze to obtain mycelium and fermentation liquor; the fungus culture medium comprises the following components: 10.0 g of glucose, 20.0 g of maltose, 20.0 g of mannitol, 10.0 g of monosodium glutamate, 3.0 g of yeast extract, 15.0 g of NaCl and KH₂PO₄ 0.5 g、MgSO₄·7H₂O0.3 g and ultrapure water were added to the reaction solution to a constant volume of 1L.

2 obtaining of extract

Separating mycelium from fermentation liquor by using gauze, continuously performing ultrasonic wall breaking on the mycelium for 3 times by using an acetone solution (containing 20% of ~ 30% of water), filtering to remove residues to obtain a crude extract of the mycelium containing the acetone and the water, performing reduced pressure concentration to remove the acetone to obtain an aqueous solution of the crude extract, adding ethyl acetate into the aqueous solution and the ethyl acetate according to the volume ratio of 1:2 to extract for 3 times to obtain an ethyl acetate crude extract, and performing concentration and evaporation to obtain 36.5 g of a mycelium extract.

3 separation and purification of Compound

The mycelium extract is mixed with 100-plus 200-mesh silica gel, and the mixture is mixed with petroleum ether: dichloromethane: methanol is used as gradient eluent, and decompression silica gel chromatographic column chromatography is carried out. The components A-E are separated by simple thin-layer chromatography analysis, combination and separation. Component C (12.7 g) was purified in dichloromethane: methanol =1:2 as gradient eluent, performing gel column chromatography (Sephadex LH-20), and mixing to obtain four subfractions C after thin layer chromatography₁-C₄. Subfraction C₃(4.9 g) by semi-preparative liquid chromatography (type 1010 ODS-A, 10X 250 mm, 5 μm): the isolation flow was 5 mL/min and the mobile phase was 55% acetonitrile with 0.1% TFA to give the indicated compound (510mg, t)_R 22.8 min)。

Said penicillium oxalicum (A), (B)Penicillium oxalicum) IBPT-6, which has been deposited at the China center for type culture Collection on 25.12.2013, address: wuhan, Wuhan university, the accession number is: CCTCC NO: m2013714.

The invention also comprises the application of the compound in the preparation of drugs for inhibiting the proliferation of human cisplatin-resistant cells and the application of the compound in the preparation of human cisplatin-resistant drugs.

The invention has the following remarkable advantages:

researches show that the penicillanicum anthrone compound has obvious activity of inhibiting the proliferation of human cisplatin-resistant cells, and no report of the activity of the compound on inhibiting the proliferation of the human cisplatin-resistant cells exists at present, so that no related medicine exists in the market.

Drawings

FIG. 1 major COSY, HMBC and NOE signals of Iso-Penicillixanthone A.

Detailed Description

The chemical structures of the compounds referred to in the examples below: