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Synthetic peptide sp4 and use thereof

a synthetic peptide and peptide technology, applied in the field of biomedicine, can solve the problems of high malignancy, prone to metastasis, and serious threat to human health and life, and achieve the effects of increasing the inhibitory activity of cdk4, and maintaining cell division

Inactive Publication Date: 2021-06-17
TAIAN CITY QIHANG BIOTECH CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present patent provides a biological product called synthetic peptide sp4, which has been shown to have anti-tumor activity. The peptide can target specific cancer cells and can be used as a single medication. It also inhibits the activity of telomerase, which is important for cell division and prevents tumor cells from infinite proliferation. The peptide can also affect the cell cycle by inhibiting the activity of cyclin dependent kinases and preventing cells from entering the S phase from the G1 phase.

Problems solved by technology

Cancer is a serious threat to human health and life.
Osteosarcoma presents invasive growth locally and grows rapidly, and it is featured by a high degree of malignancy, proneness to metastasis, poor limb salvage treatment effect, high disability and mortality, etc.
At present, in clinical practices, cytotoxic antitumor drugs are mainly used, including combined use of Paclitaxel, Cisplatinum, Methotrexate (MTX), Adriamycin (ADM), Ifosfamide (IFO), etc., and the toxic and side effects bring great harm and pain to patients.

Method used

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  • Synthetic peptide sp4 and use thereof
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  • Synthetic peptide sp4 and use thereof

Examples

Experimental program
Comparison scheme
Effect test

embodiment 1

[0033]Analysis of the Amino Acid Composition of sp4:

[0034]10.2 mg of sample was weighed and dissolved in 7 mL of 6N HCl, and hydrolyzed for 22 hours under nitrogen protection condition at 110° C. After cooling, hydrolysate was transferred to a 10 mL volumetric flask and constant-volumed. 0.2 mL of solution was taken from the flask and blew dry with 55° C. nitrogen, and 1 mL distilled water was added and dried again and repeated three times. The dried product was dissolved in 1.2 mL of deionized water (pH was adjusted with 0.02 mol / L HCl) and mixed uniformity, filtered through a 0.45 μm filter membrane, and 20 μL solution was injected in a machine (Hitachi L-8900 Amino Acid Analyzer) for testing.

[0035]Test Results:

nmol / 20 μLAmino acid ratioSerSerine4.4502.54GlyGlycine1.7531.00AlaAlanine1.7541.00IleIsoleucine3.4011.94LeuLeucine3.6152.06PhePhenylalanine3.4031.94ProProline1.7541.00

[0036]Sp4 is composed of 7 kinds amino acids of SGAILFP, as shown in FIGS. 1 A and B.

embodiment 2

[0037]Solid-phase chemical synthesis, purity testing and molecular weight of sp4:

[0038]Solid-phase chemical synthesis of sp4: The peptide Fmoc solid-phase synthesis technology was adopted, which is a process of repeatedly adding amino acids from the C-terminus to the N-terminus of the known polypeptide amino acid sequence. The process was shown in FIG. 2: firstly, covalently attaching the carboxyl group of a first amino acid at the C-terminus of the target polypeptide to the solid phase carrier (resin), and then using the amino group of this amino acid as the starting point for synthesis to have an acylation reaction with the carboxyl group of adjacent amino acid to form peptide bond. Repeating the above process until the synthesis of target polypeptide is completed, then cleaving the target polypeptide from the resin and removing the side chain protecting group contemporaneously. Finally, adding ice ether to precipitate the crude peptide, which is separated and purified by high-per...

embodiment 3

[0039]Anti-tumor in vitro screening test (method of CCK-8) was used. This technique was provided by Nanjing OGpharmaceutical science and technology Co., Ltd. and cell viability detection kits of EnoGeneCell™ Counting Kit-8 (CCK-8) were used.

[0040]Cells with living cells ratio more than 90% were taken for experiments. Cell proliferation inhibition test adopted a cell suspension at a concentration of 1×105 cells / mL after cell digestion and counting, 100 μL of cell suspension (1×104 cells per well) was added to each well of a 96-well plate; the 96-well plate was cultured at 37° C. in an incubator with 5% CO2 for 24 hours; 100 μL of corresponding drug-containing medium was added to each well, and a negative control group, a menstruum control group, and a positive control group were established contemporaneously, with 5 wells for each group. The 96-well plates were cultured at 37° C. in an incubator with 5% CO2 for 72 hours; then 10 μL of CCK-8 solution was added to each well and incubat...

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Abstract

A synthetic peptide sp4 has an amino acid sequence shown in SEQ ID NO: 1. As the sole effective ingredient in the antitumor efficacy test, it has a significant inhibitory effect on both the tumor volume and tumor weight of human osteosarcoma MG-63 in nude mice and has a significant dose-effect and time-effect relationship. There is no difference between the relative tumor growth rate T / C (%) of sp4 and that of the positive control group of Paclitaxel. The safety of sp4 is that its maximal tolerated dose for intravenous administration is 700 mg / kgBW. Sp4 has clear targets of pharmacological effects, while having efficacy in vivo, it also has an inhibitory effect on tumor telomerase, has an arrest on G1 phase of the tumor cell cycle, and inhibits the high expressions of tumor PD-L1 and CD47. The present disclosure relates to multiple uses of sp4, especially in the preparation of a class of antitumor drugs.

Description

INCORPORATION OF SEQUENCE LISTING[0001]This application contains a sequence listing submitted in Computer Readable Form (CRF). The CFR file containing the sequence listing entitled “PA128-0079-SQ_ST25.txt”, which was created on May 7, 2020, and is 483 bytes in size. The information in the sequence listing is incorporated herein by reference in its entirety.TECHNICAL FIELD[0002]The present disclosure belongs to the technical field of biomedicines, relates to a synthetic peptide sp4 (Synthetic Peptide-4) and uses thereof, in particular to uses in preparing novel anti-tumor drugs.BACKGROUND ART[0003]Cancer is a serious threat to human health and life. Osteosarcoma (OS) is a type of most common primary bone malignancy and a highly malignant mesenchymal tumor. It often occurs in adolescents below the age of 21 and children. Osteosarcoma presents invasive growth locally and grows rapidly, and it is featured by a high degree of malignancy, proneness to metastasis, poor limb salvage treatme...

Claims

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Application Information

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IPC IPC(8): C07K7/08A61P35/00
CPCC07K7/08A61K38/00A61P35/00
Inventor ZHANG, WANQINLI, YINTIANJI, XUEWENZHAO, LIMEI
Owner TAIAN CITY QIHANG BIOTECH CO
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