Synthetic peptide sp2 and application thereof

A technology for synthesizing peptides and sp2, which is applied in the field of biomedicine and can solve problems such as the blank of synthetic peptides sp2

Active Publication Date: 2019-01-18
TAIAN CITY QIHANG BIOTECH CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on the application of synthetic peptide sp2 in new anti-tumor drugs is blank

Method used

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  • Synthetic peptide sp2 and application thereof
  • Synthetic peptide sp2 and application thereof
  • Synthetic peptide sp2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Amino acid composition, solid-phase chemical synthesis, chromatographic purification, and identification by mass spectrometry

[0060] 1) Analysis of amino acid composition of sp2: Weigh 10.2mg sample, dissolve it in 7mL of 6N HCl, acid hydrolyze at 110°C under nitrogen protection for 22 hours, transfer to a 10mL volumetric flask after cooling, and constant volume. Take 0.2 mL and dry it with nitrogen at 55°C, add 1 mL of distilled water and dry it again, repeat three times. Fully dissolve with 1.2mL of deionized water (0.02mol / L HCl), and mix well. Filter with a 0.45 μM filter membrane, and inject 20 μL of the sample into the machine for testing (Hitachi L-8900 amino acid analyzer).

[0061] 2) Solid-phase chemical synthesis, purity detection and molecular weight confirmation of sp2:

[0062] The solid-phase chemical synthesis of sp2 adopts the polypeptide Fmoc solid-phase synthesis technology, which is a process of repeatedly adding amino acids, and the sequence is ...

Embodiment 2

[0066] Cell Proliferation Inhibition Assay

[0067] Cells were digested, counted, and prepared at a concentration of 1×10 5 cells / mL of cell suspension, add 100 μL of cell suspension to each well of a 96-well plate (1×10 per well 4 cells); the 96-well plate was placed at 37°C, 5%_CO 2 Cultivate in an incubator for 24 hours; add 100 μL of corresponding drug-containing medium to each well, and set up a negative control group, a vehicle control group, and a positive control group at the same time, with 5 replicate wells in each group; place the 96-well plate at 37°C, 5%_CO 2 After culturing in the incubator for 72 hours; add 10μLCCK-8 solution to each well, incubate the culture plate in the incubator for 4 hours, measure the OD value at 450nm with a microplate reader to calculate the inhibitory rate of sp2 on the BxPC-3 tumor cell line and IC 50value, the evaluation standard draws the tumor cell inhibition rate at different concentrations of the same sample to obtain a dose-ef...

Embodiment 3

[0072] In vivo evaluation of sp2 efficacy against subcutaneous human cervical carcinoma, human ovarian carcinoma, human orthotopic pancreatic carcinoma, human osteosarcoma and human poorly differentiated gastric carcinoma in nude mice

[0073] 1. Cell lines: human cervical cancer cell line SiHa, human ovarian cancer cell line A2780, human orthotopic pancreatic cancer cell line BxPC-3, human poorly differentiated gastric cancer cell line BGC-823 and human osteosarcoma cell line MG-63 All were cultured in RPMI-1640 medium containing 10% fetal bovine serum.

[0074] 2. Modeling method: subcutaneous injection of cell lines, axillary tumor formation

[0075] 3. Experimental steps:

[0076] (1) Cell preparation stage

[0077] 1: MG-63 cell line in a good recovery state, inoculated into T75 cell culture flask, 37°C, 5% CO 2 to cultivate.

[0078] 2: The medium is changed every 2-3 days, and the cell healing degree reaches about 80%. Passaging at 1:3 requires about 20 bottles.

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Abstract

The invention discloses synthetic peptide sp2 and application thereof. The amino acid sequence of the sp2 is shown by SEQ ID NO.1. The efficacy characteristics are that the synthetic peptide sp2 has obvious activity of resisting tumor growth against xenograft tumor such as nude-mouse subcutaneous human pancreatic cancer, human cervical cancer and human ovarian cancer in dose-effect and time-effectrelationships, and realizes the functions of delaying transfer and prolonging the lifetime for the nude-mouse orthotopic transplantation human pancreatic cancer and lung adenocarcinoma. The safety characteristic is that toxic reaction or death does not occur after 2,000 mg / kg BW of sp2 is applied to an animal through intravenous injection. The sp2 is also applied to: (1) the prevention and / or treatment of tumor; (2) inhibition of the proliferation and / or growth and / or invasion of tumor cells; (3) enhancement of anti-tumor immunoreaction; (4) induction of tumor cell differentiation; (5) preparation of class-A anti-tumor drugs; (6) inhibition of tumor telomerase activity; (7) control of tumor cell cycle; (8) preparation of products for regulating and controlling tumor cell cycle.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and relates to a synthetic peptide sp2 (Synthetic Peptide-2) and its application, especially the application in the preparation of new non-cytotoxic antitumor drugs. Background technique [0002] Cancer patients urgently need safe and effective anticancer drugs. Pancreatic cancer has a very high degree of malignancy and insidious onset. Metastasis to peripancreatic organs or lymph nodes, liver metastasis, and distant metastasis have occurred at the first diagnosis. Cytotoxic antineoplastic drugs, such as fluorouracil (5-FU), mitomycin (MMC), and cisplatin (ADM), have strong killing or inhibitory effects on tumor cells, but for pancreatic cancer Treatment, single-drug chemotherapy has poor curative effect and short survival time. Combination chemotherapy is often used, such as FAM (5-FU, ADM, MMC), SMF (STZ, MMC, 5-FU) schemes, etc. Since paclitaxel was launched, TC (paclitaxel + Carboplatin)...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K38/10A61P35/00
CPCA61P35/00C07K7/08A61K38/00C07K14/43522A61P35/02
Inventor 张万琴李荫田吉学文赵丽美
Owner TAIAN CITY QIHANG BIOTECH CO
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