Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

GPC3-targeted antibody-drug conjugate and preparation method and application thereof

A technology of drug conjugates and antibodies, applied in the field of medicine, can solve problems such as side reactions, cluttered electrophoretic images of antibody conjugates, and long synthetic routes of conjugated reagents.

Active Publication Date: 2019-12-17
SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI +1
View PDF11 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the methods in the prior art have long synthesis routes of coupling reagents, poor chemical stability of coupling reagents, and relatively messy electrophoresis of antibody conjugates, suggesting that there may be side reactions in the coupling process. Solve problems such as sulfhydryl exchange (reverse Michael addition reaction) during in vivo circulation

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • GPC3-targeted antibody-drug conjugate and preparation method and application thereof
  • GPC3-targeted antibody-drug conjugate and preparation method and application thereof
  • GPC3-targeted antibody-drug conjugate and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0209] Antibody preparation

[0210] The sequence of the DNA molecule of the antibody or its fragment of the present invention can be obtained by conventional techniques, such as PCR amplification or genomic library screening. In addition, the coding sequences of the light chain and the heavy chain can be fused together to form a single chain antibody.

[0211] Once the relevant sequence is obtained, the recombination method can be used to obtain the relevant sequence in large quantities. This is usually done by cloning it into a vector, then transferring it into a cell, and then isolating the relevant sequence from the proliferated host cell by conventional methods.

[0212] In addition, artificial synthesis methods can also be used to synthesize related sequences, especially when the fragment length is short. Usually, by first synthesizing multiple small fragments, and then ligating to obtain a very long fragment.

[0213] At present, the DNA sequence encoding the antibody (or fra...

Embodiment 1

[0389] Example 1. Synthesis and preparation of the compound represented by formula Ic

[0390] The substituted maleamide linker fragment molecules represented by formula Ia listed in the first aspect of the present invention can be synthesized by the method in Scheme 1. Intermediate A is obtained by reacting n-ethylene glycol with tert-butyl bromoacetate, and then with The substituted nitrofluorobenzene undergoes aromatic nucleophilic substitution to obtain intermediate B. In addition, intermediate B can also be obtained by reacting intermediate F protected by p-toluenesulfonate with substituted nitrofluorophenol. The nitro group in intermediate B is reduced to an amino group to obtain intermediate C, which is then cyclized with 2,3-dibromomaleic anhydride to obtain intermediate D, which is then substituted with arylthiophenol to obtain linker fragments Molecule E. A series of molecules F can be obtained by condensing with a dipeptide / tripeptide-PAB cytotoxic drug linker. Exam...

Embodiment 2

[0435] Example 2. Preparation and identification of anti-GPC3 antibody

[0436] The GPC3 antibody of the present invention is prepared by the following method: using a phage display antibody library, screening against GPC3 to obtain a human anti-GPC3 antibody, and sequencing and identifying the antibody with good affinity. The sequence information is as follows:

[0437] Table 2 Human anti-GPC3 antibody GPC3-6 heavy chain variable region CDR and light chain variable region CDR

[0438]

[0439] Table 3 Human anti-GPC3 antibody GPC3-6 heavy chain and light chain framework region (FR 1-4) sequence

[0440]

[0441]

[0442] Note: In the above table, aa represents the amino acid sequence, and nt represents the nucleotide sequence.

[0443] SEQ ID NO: 30GPC3-6 heavy chain variable region (VH) amino acid sequence 120aa

[0444] QVHLVQSGAEVQKPGSSVKVSCKASGGTFSSYGINWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARGSGLLPRIGGYWGKGTMVTVSS

[0445] SEQ ID NO: 29GPC3-6 heavy chain variab...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention describes an anti-glypican-3 protein (GPC3) antibody-drug conjugate. A strong cytotoxic active substance and a biological macromolecule are coupled through a novel disubstituted maleimide linker. The linker can act simultaneously with a disulfide chain, thereby greatly improving the material homogeneity of the conjugate. The conjugate prepared by the linker has high inhibitory activity on a cell strain expressing the GPC3 antigen. In addition, the invention also provides a preparation method and application of the above conjugate.

Description

Technical field [0001] The present invention relates to the field of medicine, and more specifically, to an antibody-drug conjugate targeted to Glypican 3 (GPC3), and a preparation method and application thereof. Background technique [0002] Hepatocellular carcinoma (HCC) is the fifth most common cancer and the third most common cause of death from cancer in the world. According to the American Cancer Society, hepatocellular carcinoma (HCC) accounts for approximately 75% of liver cancer cases, and often does not have symptoms until advanced liver cancer. Currently, surgery is the most effective treatment for HCC. However, the recurrence rate of tumors after radical hepatectomy is very high, and the 5-year survival rate is only 10%. In addition, because most HCC patients are diagnosed in the later stages of the disease, radical treatment, including chemotherapy, chemoembolization, resection, and proton beam therapy, may usually be ineffective. Sorafenib (Nexavar), the first cl...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/30C07K19/00C12N15/13C12N15/62C12N5/10A61K47/68A61K38/07A61P35/00
CPCA61K38/07A61K47/6851A61P35/00C07K16/303C07K2317/565C07K2319/00
Inventor 沈竞康孟韬康小强赖寿鹏马兰萍彭红丽王昕陈驎杜志彦王英于霆张永良
Owner SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com