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Tumor blood vessel and m1 type macrophage targeting peptide and use thereof

A technology of tumor blood vessels and macrophages, applied in the field of molecular biology, can solve problems such as loss, and achieve the effect of prolonging biological half-life and promoting pharmacological activity

Active Publication Date: 2021-05-14
四川正雪生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because there are no obvious and specific symptoms in the early stage of colon cancer, and most patients are often diagnosed when they are in the advanced stage and accompanied by metastatic symptoms, thus losing the opportunity of radical surgery to remove the lesions of colon cancer

Method used

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  • Tumor blood vessel and m1 type macrophage targeting peptide and use thereof
  • Tumor blood vessel and m1 type macrophage targeting peptide and use thereof
  • Tumor blood vessel and m1 type macrophage targeting peptide and use thereof

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preparation example Construction

[0039] The present invention also provides a method for preparing the above tumor blood vessel and M1 macrophage targeting peptide TCP-2, comprising the following steps:

[0040] Phage display technology was used to screen out the TCP-2 phage that specifically binds to colon cancer blood vessels, and the gene sequence encoding the TCP-2 polypeptide displayed by the phage was obtained by gene sequencing. The gene sequence was translated to obtain the TCP-2 polypeptide sequence, and the obtained polypeptide sequence was obtained. Send the peptide synthesis company to synthesize the TCP-2 polypeptide.

[0041] The present invention also provides a use of the above-mentioned tumor blood vessel and M1 macrophage targeting peptide TCP-2 in the preparation of targeted drugs for diagnosing or treating tumors.

[0042] Further, the tumor is a tumor of the digestive system, including one of colorectal cancer, gastric cancer, esophageal cancer, pancreatic cancer or liver cancer.

[0043...

Embodiment 1

[0048] Embodiment 1 prepares TCP-2

[0049] Phage display library: TCP-2 was isolated from Ph.D.TM-C7C phage display library purchased from New England Biolabs (structure shown in 1). The phage library is a combinatorial library based on random heptapeptides fused to the minor capsid protein (pIII) of the M13 phage. The randomized sequence is flanked by a pair of cysteine ​​residues. Under non-reducing conditions, two cysteines spontaneously form a disulfide cross-link, forming a phage library of cyclized peptides.

[0050] The cell culture medium is: RPMI-1640 medium containing 2g / L sodium bicarbonate, 100 units / ml penicillin and streptomycin and 10% fetal bovine serum; the culture conditions are 37°C, 5% CO 2 in the humid air.

[0051] Antibody: Rabbit anti-fd phage antibody was purchased from Sigma, rat anti-mouse CD31 monoclonal antibody, CD68 monoclonal antibody, CD80 monoclonal antibody, CD163 monoclonal antibody and mouse anti-human CD68 monoclonal antibody, CD80 mon...

Embodiment 2

[0053] Embodiment 2 constructs animal model

[0054] Experimental animals: 7-9 week-old male BALB / c mice (orthotopic colon cancer model), C57BL / 6 mice (orthotopic pancreatic cancer model) and 12-week-old ApoE - / - Mice (atherosclerosis model) were kept under conditions of humidity (50±10%), light (12 / 12 hours, light / dark cycle) and temperature (23±2°C), and all animals were divided into plastic cages Medium (four or five mice / cage), free access to drinking water and food.

[0055] Orthotopic colorectal cancer model 1: The model was established using the previously published method (see invention patent CN102471371B "homing peptide for tumor vasculature"). Colitis was induced in male BALB / c mice by continuously drinking tap water containing 3% dextran sodium sulfate (DSS) for 8 days. After fasting for 18 hours, the mice were anesthetized with sodium pentobarbital. Colon 26 cells (3×10 6 cells / 40μl / mouse) in the anus of the mouse at 2 insides. 30 minutes after instillation o...

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Abstract

The invention belongs to the technical field of molecular biology, and specifically relates to a tumor blood vessel and M1 type macrophage targeting peptide and its application. Aiming at the problem of lack of small molecule targeting peptides used for preparing drugs for diagnosis or treatment of tumors, the present invention provides a tumor vessel and M1 type macrophage targeting peptide TCP‑2 and its carrier. The small-molecule targeting peptide has the amino acid sequence shown in Seq ID NO: 1, can specifically target tumor blood vessels and M1 macrophages, and can be used to prepare targeted drugs for diagnosis or treatment of tumors, especially for use in For the treatment of gastrointestinal tumors, it provides a new option for targeted therapy of tumors.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and specifically relates to a tumor blood vessel and M1 type macrophage targeting peptide and its application. Background technique [0002] Colon cancer has become one of the most common malignant tumors worldwide, with the third highest morbidity and second highest fatality rate in the world. The pathogenesis of colon cancer is a long-term complex process caused by multiple factors, including the occurrence of hemorrhagic enteritis, benign adenomatous polyps, and then gradually develop into invasive and metastatic adenocarcinoma. Since there are no obvious and specific symptoms in the early onset of colon cancer, most patients are often diagnosed when they are in the advanced stage and accompanied by metastatic symptoms, thus losing the opportunity for radical surgery to remove the diseased tissue of colon cancer. The 5-year survival rate of stage I patients is 93.2%, while the 5-yea...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C07K1/00C12N15/11A61K47/42A61P29/00A61P35/00A61P1/00A61P1/04A61P9/10A61P19/02
CPCA61K47/42C07K7/06
Inventor 曹之宪李志杰李龙飞李明星
Owner 四川正雪生物科技有限公司
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