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Tumor specific target polypeptide and application thereof

A tumor-specific, peptide-targeting technology, applied to phage-displayed peptides and their application fields, can solve the problems of uncontrollable late-stage metastasis, drug resistance, unacceptability, etc.

Active Publication Date: 2011-05-18
INST OF RADIATION MEDICINE CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Both surgery and radiotherapy are local treatments, which cannot control advanced metastasis, and many patients cannot receive this type of treatment due to their weak constitution; while chemotherapy has some chemotherapeutic drugs with good curative effect, but due to the lack of tumor cell specificity, While killing tumor cells, it also kills a large number of bone marrow and other vigorously proliferating cells, all of which have strong side effects, and drug resistance often occurs after repeated chemotherapy, greatly reducing the efficacy of drugs

Method used

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  • Tumor specific target polypeptide and application thereof
  • Tumor specific target polypeptide and application thereof
  • Tumor specific target polypeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] 1. Establishment of nude mouse model of human A460 large cell lung cancer

[0071] 1) Take four-week-old Balb / C female nude mice (purchased from the Experimental Animal Center of Beijing Academy of Medical Sciences) and put them into an SPF grade animal room for breeding.

[0072] 2) Cultivate human non-small cell lung cancer NCI H460, take the logarithmic growth phase lung cancer cell line, digest with trypsin routinely, and adjust the cell concentration to 1x10 7 pcs / ml, take 200ul that is 2x10 6 Cells were inoculated subcutaneously under the armpit of the forelimb of each nude mouse, and the growth status of the cells after inoculation was observed. After about 5-6 days, a tumor mass was visible. After two weeks, the diameter of the tumor was about 0.3-0.5 cm, which can be used for phage screening in vivo experiment.

[0073] 2. In vivo screening of phage display peptide library in mice

[0074] (1) Culture of E.coli ER2738

[0075] Pick an inoculation loop from ...

Embodiment 2

[0120] Chemical Synthesis and Identification of Cyclic Heptapeptide:

[0121] From the results of the sequencing of the cyclic heptapeptide, the amino acid sequence of the cyclic heptapeptide was deduced to be ACPLSHSLIC. .The PLSHSLI in the middle of CPLSHSLIC plays a targeting role. Synthesized and labeled with FITC (fluorescein isothiocyanate) as the detection material. The cyclic heptapeptide was purified and identified by HPLC. The results showed that the synthetic cyclic heptapeptide had a purity of more than 98%, and its molecular weight was consistent with the theoretical value.

Embodiment 3

[0123] The polypeptide is connected with the nanomaterial polyamide-amine dendrimer (PAMAM) by chemical cross-linking method, and the PAMAM is connected with fluorescein isothiocyanate (FITC) as a detection label.

[0124] 1. Acetylation of fourth-generation PAMAM

[0125] Dissolve 1 g of PAMAM in 120 ml of anhydrous methanol, and add 0.35 g of triethylamine. Dissolve 0.21g of acetyl anhydride in 60ml of anhydrous methanol, and add dropwise to the above solution, and stir overnight at room temperature. Anhydrous methanol was evaporated in rotary vacuum and the residue was dissolved in water. Dialyze through a dialysis bag to remove unreacted small molecular substances, and freeze-dry after dialysis.

[0126] 2. Connect to FITC

[0127] Weigh 0.4 g of the acetylated PAMAM and dissolve in 40 ml of DMSO, dissolve 38.9 mg of FITC in 8 ml of DMSO, add dropwise to the PAMAM solution, and stir overnight. The reaction solution was dialyzed to remove unreacted small molecular subst...

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Abstract

The invention relates to a polypeptide capable of specific tumor-binding. The amino acid sequence of the polypeptide is shown as SEQ ID No.1. According to the invention, phages specifically binding with lung cancer are obtained by means of a phage display in vivo screening method to determine sequence in order to synthesize the specific binding polypeptide. The polypeptide can achieve excellent in vitro binding with a plurality of tumor cells subsequent to the conjunction with poly(amidoamine) dendrimer nano material. As verified by in vitro experiments, the target polypeptide has a very hightumor targeting effect and can be widely used as a targeting agent in diagnostic imaging and targeted treatment of tumors.

Description

technical field [0001] The invention belongs to the technical field of protein polypeptides, and relates to polypeptides displayed by phages and applications thereof. More specifically, it is a polypeptide that can specifically bind to tumor cells and its preparation method and application. Background technique [0002] Tumor is one of the most common cancers in the world. In many advanced countries, the death rate of lung cancer ranks first among all cancer deaths. Currently, lung cancer treatment mainly includes surgery, radiotherapy and chemotherapy. Both surgery and radiotherapy are local treatments, which cannot control advanced metastasis, and many patients cannot receive this type of treatment due to their weak constitution; while chemotherapy has some chemotherapeutic drugs with good curative effect, but due to the lack of tumor cell specificity, While killing tumor cells, it also kills a large number of bone marrow and other vigorously proliferating cells, all of ...

Claims

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Application Information

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IPC IPC(8): C07K7/06C12N15/11A61K38/08G01N33/574A61P35/00
Inventor 刘鉴峰褚丽萍刘金剑王彦王月英吴红英李德冠冯丽娜
Owner INST OF RADIATION MEDICINE CHINESE ACADEMY OF MEDICAL SCI
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