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Preparation method of nettle cyclopeptide in cockscomb seeds and application of nettle cyclopeptide in antitumor drugs

A technology of Urticaceae and cyclic peptides, applied in the field of pharmaceutical technology, anti-tumor compounds, and natural medicinal chemistry, can solve problems such as the mechanism of anti-tumor action of Moroidin, and achieve good anti-tumor activity, wide clinical application prospects, and dosage forms and the effect of diversification of medication

Active Publication Date: 2022-05-13
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, there is no report on the isolation of the Urticaceae type cyclic peptide Moroidin from Cockscomb, nor is there any report on the anti-tumor effect of Moroidin and its related mechanism of action

Method used

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  • Preparation method of nettle cyclopeptide in cockscomb seeds and application of nettle cyclopeptide in antitumor drugs
  • Preparation method of nettle cyclopeptide in cockscomb seeds and application of nettle cyclopeptide in antitumor drugs
  • Preparation method of nettle cyclopeptide in cockscomb seeds and application of nettle cyclopeptide in antitumor drugs

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The preparation method of embodiment 1 Urticaceae type cyclic peptide Moroidin (1) (flow chart sees figure 1 shown).

[0030] Take dried cockscomb flower seeds (30kg), heat and reflux extraction with 10 times the volume of 70% ethanol after crushing, extract three times (30L×3), each time for 3 hours, combine the extracts, and concentrate under reduced pressure to obtain the total Extract (1.05kg). Suspend the total extract in water, extract continuously with petroleum ether, ethyl acetate, n-butanol respectively, extract three times with equal volume, reclaim the solvent to obtain petroleum ether part (33g), ethyl acetate part (60g), n-butanol Part (Fr.BuOH, referred to as Fr.B, 413g) and the water part after extraction.

[0031] The fraction of n-butanol (Fr.B, 413g) was chromatographed by D101 macroporous adsorption resin column, with methanol / water system (10:90, 30:70, 50:50, 70:30, 90:10, 0:100 , v / v) gradient elution, wherein 50% of the methanol / water system (...

Embodiment 2

[0040] Cytotoxic effect of the Urticaceae type cyclic peptide Moroidin prepared in Example 2 on tumor cells

[0041] Experimental method: using thiazolyl blue (3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide, MTT) experiment, plant cyclic peptide Moroidin (concentration gradient is 5 μM, 10 μM , 20 μΜ, 40 μΜ, 80 μΜ) to treat lung cancer cells A549, H1299, breast cancer cells MCF7, glioma cells U87, U251, and colorectal cancer cells HCT116, and analyze the in vitro anti-tumor activity of plant cyclic peptide Moroidin. The specific steps are: (1) Cell sorting: Tumor cells in good logarithmic growth phase were inoculated into 96-well plates at an inoculation concentration of 5000 cells / well, and cultured in a conventional cell incubator for 12 hours. (2) Administration: The culture medium was discarded, and the experimental groups were given different concentrations of plant cyclic peptide Moroidin (5 μM, 10 μM, 20 μM, 40 μM, 80 μM), and 5 replicate wells were set for ...

Embodiment 3

[0043] Example 3 Effect of Urticaceae Cyclic Peptide Moroidin on Long-term Proliferation of Tumor Cells

[0044] Experimental method: The plant cyclic peptide Moroidin of the present invention acts on lung cancer cell A549 and breast cancer cell MCF7 at different concentrations, and detects the long-term growth of tumor cells through a clone formation experiment.

[0045] The tumor cells in the logarithmic growth phase were evenly seeded in 6-well plates, with 500 cells per plate. After the cells adhered to the wall, they were treated with different concentrations of Moroidin for 7-10 days, and the cell colonies were stained with 0.1% crystal violet solution for 2 minutes. The background color was eluted with PBS, and then photographed. Use the Image Tool software to calculate the number of clones, and calculate the cloning rate with the following formula: cloning rate=(forming the number of clones / the number of planted cells)×100%

[0046] Experimental results: as attached ...

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Abstract

The invention discloses a preparation method of nettle cyclopeptide in cockscomb seeds and application of the nettle cyclopeptide in antitumor drugs. The nettle cyclic peptide compound Moroidin is extracted from cockscomb seeds for the first time, and a new plant source for obtaining the cyclic peptide and a preparation method are provided. In-vitro anti-tumor experiments show that Moroidin has a toxic effect and an anti-proliferation effect on various malignant tumor cells for the first time and can block tumor cells in a G2 / M phase so as to induce apoptosis of the tumor cells, so that Moroidin has good anti-tumor activity and has potential application value in preparation of drugs for treating tumor-related diseases.

Description

technical field [0001] The invention belongs to the fields of natural medicinal chemistry, pharmaceutical technology and anti-tumor compounds, and in particular relates to a preparation method of extracting Urticaceae type cyclic peptide Moroidin from Cockscomb flower seeds, and its application as a tumor cell proliferation inhibitor and in the preparation of anti-tumor drug application. Background technique [0002] Malignant tumors are a serious disease that seriously endangers human health, and the morbidity and mortality are increasing year by year. Although new treatment methods such as gene therapy and immunotherapy have entered clinical practice with the advancement of science and technology, chemical drugs are still an important means of treating malignant tumors so far. Antineoplastic drugs come from a wide range of sources, and natural products derived from plants have become an important source of antineoplastic drugs because of their diverse structures and defin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K1/14C07K1/22C07K1/20C07K1/16A61K38/08A61P35/00
CPCC07K7/06A61P35/00A61K38/00Y02A50/30
Inventor 张爱霞范君婷徐筱雅刘尚明金阳蒋南都述虎
Owner NANJING MEDICAL UNIV
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