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Polypeptide-albumin coupling drug and preparing method and application thereof

An albumin and albumin-binding technology, applied in the field of polypeptide-albumin conjugated drug preparation, can solve the problems of toxicity, high production and identification costs, limited molecular weight and passive transport ability to penetrate epithelial cell membranes, etc. Lethal, strong specific effect

Inactive Publication Date: 2016-03-30
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, monoclonal antibody as a targeted drug carrier has the following disadvantages: its large molecular weight limits the passive transport ability to penetrate the epithelial cell membrane; non-specific absorption to the liver and reticuloendothelial system leads to its dose-limiting toxicity; production and identification costs higher

Method used

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  • Polypeptide-albumin coupling drug and preparing method and application thereof
  • Polypeptide-albumin coupling drug and preparing method and application thereof
  • Polypeptide-albumin coupling drug and preparing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Synthesis of Polypeptide PepB Specifically Binding to Epithelial Cell Adhesion Molecules

[0032] The amino acid sequence of the polypeptide PepB: VRRDAPRFSMQGLDACGGNCNK-BMPA, wherein BMPA is a 3-maleimide group, and the amino acid sequence of the polypeptide PepB labeled with fluorescein isothiocyanate (FITC) is FITC-VRRDAPRFSMQGLDACGGNCNK-BMPA, as shown in Table 1 shown. The peptide was synthesized according to the amino acid sequence shown (synthesized by Shanghai Keyept Biotechnology Co., Ltd., with a purity of 98%), and a mother solution of appropriate concentration was prepared with PBS buffer before the experiment.

[0033] Table 1 Peptide sequence

[0034] Peptide name

[0035] It should be noted that the function of FITC is to use the polypeptide for the characterization of fluorescence detection means, which is a commonly used technical means and has no substantial impact on the function of the polypeptide of the present invention.

Embodiment 2

[0036]The verification of embodiment 2EpCAM positive and negative cell lines

[0037] 1. Cell material used

[0038] The SK-BR-3 cell line was used as EpCAM-positive cells to be verified, and the HL-60 cell line was used as EpCAM-negative cells to be verified.

[0039] 2. Specific method

[0040] 1) Peptide dissolution: Dissolve the peptide FITC-Pep7-2 powder in PBS buffer to a concentration of 1 mg / mL mother solution, then vortex for 3 minutes to ensure that the peptide is fully dissolved and dispersed, and store at 4°C for later use.

[0041] 2) Cell counting: culture SK-BR-3 and HL-60 cells according to the cell culture method. For the adherent cell SK-BR-3, when it grows to 80-90% of the culture dish and connects into a network, Digest and centrifuge it, discard the culture medium, add PBS to resuspend, gently pipette and evenly count; for the suspension cell HL-60, when it grows to 80-90% of the culture dish, centrifuge it and discard the culture medium , add PBS to re...

Embodiment 3

[0048] Example 3 Targeting Verification of Bifunctional Polypeptide PepB

[0049] 1. Cell lines used

[0050] The verified SK-BR-3 cell line consistent with the report was regarded as EpCAM-positive cells, and the HL-60 cell line was regarded as EpCAM-negative cells.

[0051] 2. Specific method

[0052] 1) Polypeptide dissolution: Dissolve the peptide FITC-Pep7-2 and peptide FITC-PepB powders in PBS buffer to a concentration of 1 mg / mL mother solution, then vortex and shake for 3 minutes to ensure that the peptides are fully dissolved and dispersed, and store at 4°C for later use.

[0053] 2) Cell counting: culture SK-BR-3 and HL-60 cells according to the cell culture method. For the adherent cell SK-BR-3, when it grows to 80-90% of the culture dish and connects into a network, Digest and centrifuge it, discard the culture medium, add PBS to resuspend, gently pipette and evenly count; for the suspension cell HL-60, when it grows to 80-90% of the culture dish, centrifuge it a...

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Abstract

The invention provides a polypeptide-albumin coupling drug for treating cancer. Polypeptide can be specifically bonded with epithelial cell adhesion molecules (EpCAM), and Michael addition reaction can be conducted between the terminal modification 3-maleimide group (BMPA) and free cysteine of human serum albumin; the polypeptide-albumin coupling drug is formed through chemical coupling of an albumin binding type chemotherapy drug and polypeptide, and preferably, the albumin binding type chemotherapy drug is Abraxane. The polypeptide-albumin coupling drug has high specificity, has extremely high cancerous cell killing ability, provides a feasible method and technical means for targeted therapy of cancer, and has significant value in targeted therapy of tumor.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a polypeptide-albumin conjugate drug, its preparation method and application. Background technique [0002] Tumor is one of the diseases that seriously endanger human health. The incidence and mortality of tumors in my country are increasing year by year. Tumors are divided into two categories, benign tumors and malignant tumors, while malignant tumors of epithelial origin are also called carcinomas. In my country, an average of nearly 8,500 people are diagnosed with cancer every day, 6 people are diagnosed with cancer every minute, and 1 person dies of cancer every 7 to 8 people. The top three cancers in the country are lung cancer, breast cancer and gastric cancer. Statistics show that in the past two years, the incidence of female cancer has increased significantly, especially breast cancer, which ranks first among female malignant tumors, with about 210,000 ...

Claims

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Application Information

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IPC IPC(8): C07K14/00C07K14/76C07K1/34C07K1/02A61K31/337A61K47/48A61P35/00
CPCC07K14/00A61K31/337C07K14/76
Inventor 王琛侯静菲杨延莲于越
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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