Specific HER2 protein targeted polypeptide and application thereof

A specific and targeted technology, applied in applications, specific peptides, immunoglobulins, etc., can solve problems such as large toxic and side effects, and achieve the effects of small molecular weight, strong selectivity and high purity

Active Publication Date: 2016-06-22
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

While killing tumor cells, it also kills normal ce...

Method used

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  • Specific HER2 protein targeted polypeptide and application thereof
  • Specific HER2 protein targeted polypeptide and application thereof
  • Specific HER2 protein targeted polypeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Synthesis of a polypeptide specifically targeting HER2 protein

[0037] 1. Experimental instruments and materials

[0038] Dimethylformamide (DMF), piperidine, resin, dichloromethane (DCM), ninhydrin reagent (ninhydrin, vitamin C, phenol), tetramethyluronium hexafluorophosphate (HBTU), six Hydropyridine (piperidine), triisopropylsilane (TIS), ethanedithiol (EDT), anhydrous ether, trifluoroacetic acid (TFA), N-methylmorpholine (NMM), methanol, various amino acids , Peptide solid-phase synthesis tube.

[0039] 2. Solution preparation

[0040] Deprotection solution - hexahydropyridine: DMF = 1:4

[0041] Reaction solution——NMM:DMF=1:24

[0042] Lysis solution - TFA (92.5%), TIS (2.5%), EDT (2.5%), H 2 O(2.5%)

[0043] Ninhydrin Test Solution—Ninhydrin: Vitamin C: Phenol = 1:1:1

[0044] 3. Experimental steps

[0045] Weigh the resin and put it into the peptide solid-phase synthesis tube (hereinafter referred to as the reactor), and add an appropriate amou...

Embodiment 2

[0050] Embodiment 2 Immunofluorescence method detects the binding effect of SEQIDNO: 1 and 2 polypeptides to human HER2 positive breast cancer cells

[0051] 1. Experimental method

[0052] The human breast cancer HER2 high-expressing cell line SKBR3 was suspended in RPMI1640 culture medium containing 10% heat-inactivated fetal bovine serum, and seeded in 3 confocol small dishes at a density of 3000-5000 cells / dish. After cultivating for 24 hours, suck the medium in the small dish, and then add respectively 200 μL of medium containing SEQIDNO:1 and SEQIDNO:2 polypeptides labeled with fluorescein isothiocyanate (FITC) (FITC). The culture medium of PBS (phosphate buffer pH 7.4) equal to the amount of the polypeptide, and the nuclei were stained with hoechst reagent, and used in a 1:200 dilution. Incubate in an ice bath protected from light for 30 minutes. After washing with PBS and repeating the washing 3 times, 200 μL PBS was added, and the fluorescent signal was observed usi...

Embodiment 3

[0055] Embodiment 3 Immunofluorescence method detects the binding effect of SEQIDNO: 1 and 2 polypeptides to human HER2-negative breast cancer cells

[0056] 1. Experimental method

[0057] The HER2-negative breast cancer cell line 468 was suspended in H-DMEM medium containing 10% heat-inactivated fetal bovine serum, and seeded in 3 confocol small dishes at a density of 3000-5000 cells / dish. After cultivating for 24 hours, suck the medium in the small dish, and then add respectively 200 μL of medium containing SEQIDNO:1 and SEQIDNO:2 polypeptides labeled with fluorescein isothiocyanate (FITC) (FITC). The culture medium of PBS (phosphate buffer pH 7.4) equal to the amount of the polypeptide, and the nuclei were stained with hoechst reagent, and used in a 1:200 dilution. Incubate in an ice bath protected from light for 30 minutes. After washing with PBS and repeating the washing 3 times, 200 μL PBS was added, and the fluorescent signal was observed using a confocal laser micro...

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Abstract

The invention provides specific HER2 (human epidermal growth factor receptor 2) protein targeted polypeptide. The polypeptide is a mutant obtained with pertuzumab 46-65th amino acid as a template, can be specifically combined with HER2 high-expression positive breast cancer cells, and has the higher affinity than pertuzumab. The invention further provides a derivative of the polypeptide. The derivative is a bivalent or polyvalent formed from the polypeptide. The polypeptide has the character of targeting HER2 positive cancer cells, and is high in selectivity. The polypeptide can be prepared through a chemical synthesis method, is high in purity, small in molecular weight, strong in specificity, free of immunogenicity, safe and reliable, and can be used for cancer targeted therapy or cancer targeted molecular imaging and diagnosis.

Description

technical field [0001] The invention relates to the field of medicinal chemistry, in particular to a polypeptide specifically targeting HER2 protein and its application. Background technique [0002] According to the "World Cancer Report", it is predicted that the global cancer cases will show a rapid growth trend, from 14 million in 2012 to 19 million in 2025, and will reach 24 million in 2035. In 2012, there were 14 million new cancer cases and about 8.2 million cancer deaths worldwide. Among them, 3.07 million new cancer patients and about 2.2 million people died of cancer in China, accounting for 21.9% and 26.8% of the global total respectively. The most common types of cancer causing cancer deaths worldwide are: lung cancer (1.59 million deaths), liver cancer (745,000 deaths), stomach cancer (723,000 deaths), colorectal cancer (694,000 deaths), breast cancer (521,000 deaths) deaths), esophageal cancer (400,000 deaths). Among them, breast cancer is the most common can...

Claims

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Application Information

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IPC IPC(8): C07K16/32C12N15/13C12N15/63A61K47/42A61K47/48A61K51/10A61K49/00A61P35/00G01N33/574
Inventor 杨小亮耿令令方巧君
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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