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A kind of polypeptide targeting pd-l1 and its application

A PD-L1, targeting technology, applied in the field of molecular biology and medicine, can solve the problems of cumbersome preparation of antibody drugs, poor in vitro stability, weak penetration, etc., and achieves good application prospects and clinical guiding significance, small molecular weight, low cost effect

Active Publication Date: 2022-02-08
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

[0009] Antibody drugs have problems such as cumbersome preparation, poor in vitro stability, large molecules, difficult labeling, weak penetration, post-translational modification and high cost, which limit their further application

Method used

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  • A kind of polypeptide targeting pd-l1 and its application
  • A kind of polypeptide targeting pd-l1 and its application
  • A kind of polypeptide targeting pd-l1 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1 Construction and screening of PD-L1 targeting polypeptide library

[0067] (1) Experimental instruments and materials

[0068] N-methylmorpholine (NMM), piperidine, trifluoroacetic acid (TFA), dichloromethane (DCM), ninhydrin, vitamin C, phenol, tetramethyluronium hexafluorophosphate (HBTU), hexahydro Pyridine, triisopropylsilane (TIS), ethanedithiol (EDT), N,N dimethylformamide (DMF), anhydrous ether, resin, methanol, various Fmoc protected amino acids, streptavidin Magnetic beads (MB-Streptavidin), biotin labeling kit, peptide synthesis tube, shaker, vacuum water pump, rotary evaporator, laser confocal microscope (ZEISS LSM 710), the above reagents and materials were obtained from commercial sources.

[0069] (2) Solvent preparation

[0070] The preparation of the deprotection solvent is hexahydropyridine: N, N dimethylformamide = 1:4;

[0071] The preparation of the reaction solution is N-methylmorpholine: N, N dimethylformamide = 1:24;

[0072] Prepara...

Embodiment 2

[0091] Example 2 Detection of affinity between PDP-1 polypeptide and PD-L1 protein

[0092] The affinity between the PDP-1 polypeptide and the PD-L1 protein was detected by the surface plasmon resonance (SPRi) method, as follows:

[0093] Spot 1mg / mL PDP-1 polypeptide and 1×PBS on the chip, incubate overnight at 4°C under humid conditions, then wash with 10×PBS for 10 minutes, then wash with 1×PBS for 10 minutes, and finally wash with deionized water for 2 minutes 10 min each time, immersed in 1×PBS containing 5% skimmed milk, incubated overnight at 4°C, then washed with 10×PBS for 10 min, 1×PBS for 10 min, and finally washed twice with deionized water, each time 10min, blow dry with nitrogen, put the chip on the machine (Plexera HT surface plasmon resonance imaging system).

[0094] The mobile phase was sequentially passed through 1×PBS, 2×PBS, 0.625 μg / mL, 1.25 μg / mL, 2.5 μg / mL, 5 μg / mL and 10 μg / mL human PD-L1 purified protein, and the SPRi signal was recorded and analyz...

Embodiment 3

[0096] Example 3 Preparation of PDP-1 Polypeptide Conjugates

[0097] This embodiment takes the preparation of polypeptide fluorescent conjugates as an example to illustrate the preparation of polypeptide PDP-1 conjugates, as follows:

[0098] Weigh 300 mg of wang-Glu resin, cycle according to the solid-phase peptide synthesis procedure, first deprotect, and then add a certain amount of Gln, Asp, Glu, Tyr, Tyr, Thr, Asn and an equal amount of HTBU in sequence to react. After the coupling reaction, deprotect and wash. Add ε-aminocaproic acid for reaction, and react overnight with FITC and peptide beads in a solution with a ratio of 1:5:7 of pyridine / N,N dimethylformamide / dichloromethane. After the reaction, wash. Finally, the lysate was lysed into the above-mentioned resin, the side chain protecting group was removed, and vacuum-dried to obtain the crude polypeptide fluorescent conjugate.

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Abstract

The invention relates to the technical fields of molecular biology and medicine, in particular to a polypeptide targeting PD-L1 and its application. The present invention provides a polypeptide targeting PD-L1, its general formula is NX 5 x 4 x 3 EX 2 x 1 g. The polypeptide of the present invention has high affinity and specificity to PD-L1, can specifically and efficiently target PD-L1 positive tumor cells, and can be used as a polypeptide molecular probe to detect the expression of PD-L1 in tumor cells, and to Real-time monitoring of the efficacy of immunotherapy and screening of patients suitable for immunotherapy; it can also be used as a targeting polypeptide for targeted therapy and imaging of tumors with high expression of PD‑L1, and has broad application prospects.

Description

technical field [0001] The present invention relates to the technical fields of molecular biology and medicine, in particular to a polypeptide targeting programmed death ligand-1 (PD-L1), products prepared from the polypeptide or containing the polypeptide and applications thereof. Background technique [0002] In recent years, tumor immunotherapy has developed into an important anti-tumor treatment method. Different from surgical treatment, chemical drug treatment, radiation therapy and targeted therapy, tumor immunotherapy is a treatment that activates the body's own immune system to fight against tumors. , to provide the possibility to overcome malignant tumors. Tumor immunotherapy has a variety of therapeutic strategies, including non-specific immune stimulants, tumor vaccines, adoptive immune cell therapy, and monoclonal antibody therapy. Due to the great heterogeneity and genetic instability of tumors, its pathogenesis is complex, and it is difficult to achieve the id...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C12N15/11G01N33/574G01N33/576G01N33/68A61K38/08A61K47/42A61K47/64A61K49/00A61K49/14A61K49/18A61P35/00A61P37/02
CPCC07K7/06G01N33/57484G01N33/5767G01N33/6893A61K47/42A61K47/64A61K49/0056A61K49/14A61K49/1812A61P35/00A61P37/02G01N2333/70532G01N2800/102G01N2800/22G01N2800/323G01N2800/324A61K38/00
Inventor 胡志远钱怡霞王月华王蔚芝
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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