阅读说明：本技术 从棕色扁海绵中提取的环肽类化合物及其制备方法与用途 (Cyclic peptide compound extracted from Phakellia fusca and preparation method and application thereof ) 是由 林厚文 王淑萍 吴莹 颜益珍 孙凡 刘丽云 于 2019-02-26 设计创作，主要内容包括：本发明公开了一种从棕色扁海绵中提取的环肽类化合物,化学结构式如下：<Image he="642" wi="700" file="DDA0001978188820000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>本发明的环肽类化合物制备方法简单,对HePG2细胞抑制活性显著。本发明为研究和开发新抗肿瘤药物提供了新的先导化合物,为开发利用我国海洋药用资源提供了科学依据。(The invention discloses a cyclic peptide compound extracted from Phakellia fusca, which has the following chemical structural formula: the cyclic peptide compound provided by the invention is simple in preparation method and has obvious inhibitory activity on HePG2 cells. The invention provides a new lead compound for researching and developing new anti-tumor drugs and provides scientific basis for developing and utilizing marine medicinal resources in China.)

1. A cyclic peptide compound extracted from Phakellia fusca is characterized in that: the chemical structural formula is as follows:

2. a process for the preparation of cyclic peptides extracted from Phakellia fusca according to claim 1, comprising the steps of:

crushing Phakellia fusca, percolating and extracting for 8 times by using 95% ethanol, mixing extracting solutions, concentrating under reduced pressure, and concentrating to obtain an extracting solution; the concentration of the Phakellia fusca dissolved in the ethanol is 0.02-0.04 g/mL;

extracting the extracting solution with equal volume of ethyl acetate for three times to obtain ethyl acetate layer extract; suspending the ethyl acetate layer extract in 90% methanol aqueous solution, extracting with isovolumetric petroleum ether for three times, combining the lower layer solutions, diluting with water, extracting with isovolumetric dichloromethane for three times, combining the extracts, and concentrating under reduced pressure to obtain dichloromethane layer extract;

subjecting the dichloromethane layer extractive solution to Sephadex LH-20 gel column chromatography with 50% CH₂Cl₂Eluting with MeOH eluent, enriching with large molecular weight compounds by mass spectrometry, and separating by ODS medium-pressure column chromatography with MeOH/H₂Performing gradient elution, performing mass spectrum tracking and positioning analysis to obtain fine fractions containing the cyclic peptide compounds with large molecular weight, and finally separating the fractions by mass spectrum-guided semi-preparative high performance liquid chromatography to obtain the cyclic peptide compounds.

3. Use of cyclic peptides extracted from Phakellia fusca according to claim 1 in the preparation of an anti-tumor medicament.

4. The cyclic peptide compound extracted from Phakellia fusca according to claim 3, wherein the cyclic peptide compound is extracted from Phakellia fusca, and the cyclic peptide compound is extracted from Phakellia fusca and used for preparing an antitumor drug, wherein the cyclic peptide compound is extracted from Phakellia fusca according to the following formula: the tumor is liver cancer.

5. A pharmaceutical composition characterized by: cyclic peptides extracted from Phakellia fusca according to claim 1 as an active ingredient.

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a cyclic peptide compound extracted from Phakellia fusca, and a preparation method and application thereof.

Background

Sponges are a rich source of bioactive peptides, including linear peptides, depsipeptides, and cyclic peptides. Cyclic peptides of sponge origin are widely studied for their structural diversity, excellent stability and potent biological activity. In recent years, peptides have been receiving increasing attention as therapeutic agents, more than 60 peptide drugs have entered the market, and hundreds of new therapeutic peptides are in preclinical and clinical development stages, such as marine peptide ascotides that have been approved by FDA for pain treatment and dolastatin-derived drugs for cancer treatmentThus, marine-derived peptides have been the focus of research by natural product chemists.

Disclosure of Invention

The first purpose of the invention is to provide a cyclic peptide compound extracted from Phakellia fusca.

The second purpose of the invention is to provide a preparation method of the cyclic peptide compound extracted from the Phakellia fusca.

The third purpose of the invention is to provide the application of the cyclic peptide compound extracted from the Phakellia fusca in preparing the anti-tumor medicine.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

the invention provides a cyclic peptide compound extracted from Phakellia fusca, which is a novel cyclic peptide compound separated from marine invertebrate Phakellia fusca collected from the territorial waters near Hsisha Islands, and has the molecular formula C₃₉H₅₃N₇O₈The chemical structural formula is as follows:

the second aspect of the invention provides a preparation method of the cyclic peptide compound extracted from the Phakellia fusca, which comprises the following steps:

crushing Phakellia fusca, percolating and extracting for 8 times by using 95% ethanol, mixing extracting solutions, concentrating under reduced pressure, and concentrating to obtain an extracting solution; the concentration of the Phakellia fusca dissolved in the ethanol is 0.02-0.04 g/mL;

extracting the extracting solution with equal volume of ethyl acetate for three times to obtain ethyl acetate layer extract; suspending the ethyl acetate layer extract in 90% methanol aqueous solution, extracting with isovolumetric petroleum ether for three times, combining the lower layer solutions, diluting with water, extracting with isovolumetric dichloromethane for three times, combining the extracts, and concentrating under reduced pressure to obtain dichloromethane layer extract;

subjecting the dichloromethane layer extractive solution to Sephadex LH-20 gel column chromatography with 50% CH₂Cl₂Eluting with MeOH eluent, enriching with large molecular weight compounds by mass spectrometry, and separating by ODS medium-pressure column chromatography with MeOH/H₂Performing gradient elution, performing mass spectrum tracking and positioning analysis to obtain fine fractions containing the cyclic peptide compounds with large molecular weight, and finally separating the fractions by mass spectrum-guided semi-preparative high performance liquid chromatography to obtain the cyclic peptide compounds.

The third aspect of the invention provides application of the cyclic peptide compound extracted from the Phakellia fusca in preparing an anti-tumor medicament, wherein the tumor is liver cancer.

In a fourth aspect, the invention provides a pharmaceutical composition comprising the cyclic peptide compound as an active ingredient.

The cyclic peptide compound of the invention can show strong inhibitory activity to human liver cancer cell HePG2, has close relation with tumor proliferation and apoptosis, and can be used for preparing antitumor drugs.

Due to the adoption of the technical scheme, the invention has the following advantages and beneficial effects:

the medicine source material Phakellia fusca (Phakellia fusca) of Demospongiae of Halichondridae of Axinellidae, the cyclic peptide compound provided by the invention is separated from marine invertebrate Phakellia fusca collected from the territorial waters near Hsisha Islands. The cyclic peptide compound provided by the invention is simple in preparation method and has obvious inhibitory activity on HePG2 cells. The invention provides a new lead compound for researching and developing new anti-tumor drugs and provides scientific basis for developing and utilizing marine medicinal resources in China.

Drawings

FIG. 1 is a MS/MS fragment detection scheme of cyclic peptide compounds of the present invention.

FIG. 2 is a high-grade Marfey's analysis diagram of the cyclic peptide compounds of the present invention by LC-MS.

FIG. 3 is a graph showing the inhibition rate of the cyclic peptide compounds of the present invention against HePG2 cells at various concentrations.

Detailed Description

In order to more clearly illustrate the invention, the invention is further described below in connection with preferred embodiments. It is to be understood by persons skilled in the art that the following detailed description is illustrative and not restrictive, and is not to be taken as limiting the scope of the invention.

The invention relates to a novel cyclic peptide compound which is separated from marine invertebrate Phakelliafusca collected from the sea area near the Hsisha Islands in China and has the cytotoxicity of HePG 2.