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

Antibody prodrug conjugate and preparation and use thereof

A technology of antibody prodrugs and conjugates, which is applied in the field of preparation of antibody prodrug conjugates, can solve the problems of bystander toxicity, poor metabolic stability, etc., achieve good tumor treatment effect, low toxicity and side effects, and enhance tumor tissue The effect of penetration

Active Publication Date: 2019-06-28
PEKING UNIV
View PDF2 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this tumor-released ADC drug remains problematic
Due to the poor metabolic stability of most tumor microenvironment-responsive linkers, coupled with the heterogeneity of the tumor microenvironment, many release-type ADC drug molecules will release small molecule drugs early
These small molecule drugs diffuse to adjacent normal cells, causing bystander toxicity

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
  • Antibody prodrug conjugate and preparation and use thereof
  • Antibody prodrug conjugate and preparation and use thereof
  • Antibody prodrug conjugate and preparation and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1. Synthesis of doxorubicin prodrug (TCO-Dox-Mal) protected by trans-cyclooctene in response to tumor microenvironment pH The synthetic route of TCO-Dox-Mal is as follows:

[0044]

[0045]

[0046] (1) Synthesis of compound 1

[0047]Add cyclooctene-2-alcohol ((Z)-cyclooct-2-en-1-ol) (8.5g, 67mmol), methyl benzoate (9.2g, 67mmol), 500mL to 1L quartz bottle Anhydrous ether and 1000mL n-hexane. The reaction solution was subjected to light reaction for 24 hours, the wavelength of the light source was 254nm, and the intensity was 300W. During the photoreaction, the reaction system was circulated through a peristaltic pump, and the product was enriched through a silver nitrate silica gel column (the mass fraction ratio of silver nitrate and silica gel was 25:200), and the flow rate of the peristaltic pump was 18 mL / min. After the reaction, the silver nitrate silica gel was taken out, 500 mL of ammonia water was added and stirred for 5 min, and then extracted...

Embodiment 2

[0054] Example 2. Preparation of Antibody Prodrug Conjugate Ab-Dox-TCO and Determination of DAR

[0055] For the preparation process of antibody prodrug conjugates, see figure 1 . Dissolve 10 mg of trastuzumab monoclonal antibody in 10 mL of PBS solution, and then add DTT (33.3 μL, 3.33 μmol) for reduction at 37° C. for 30 min. After the reaction, the reduced trastuzumab monoclonal antibody was desalted with a PD 10 desalting column, and then centrifugally concentrated to 10 mg / mL for use. Prepare 100 mM TCO-Dox-Mal DMSO solution, take 33.3 μL and add it to the above 10 mg / mL reduced monoclonal antibody solution. Then the reaction system was carried out at 4° C. for 1 h. After the end, repeated desalting and concentration were carried out to finally obtain the antibody prodrug conjugate. Wherein, the concentration of the antibody prodrug conjugate was measured by A280 on the Nanodrop (1 mg / mL=1.4AU). The concentration of Dox is obtained by absorbing light at 495nm on the ...

Embodiment 3

[0056] Example 3. SDS-PAGE Characterization of Antibody Prodrug Conjugates

[0057] Take 16 μL of 10 mg / mL antibody prodrug conjugate Ab-Dox-TCO and antibodies of the same volume and concentration, add 4 μL of SDS-PAGE loading buffer respectively, and then boil the sample at 95°C for 30 minutes to fully denature the protein. Then take 10 μL of the boiled sample for SDS-PAGE electrophoresis. Electrophoresis conditions are 12% polyacrylamide gel, 100V voltage for 1h. After the end, the gel was stained with Coomassie Brilliant Blue, and after decolorization, it was developed with ChemiDoc XRS+molecular imager (molecular imager), as shown in figure 2 shown. The channels used are Comassie channel and fluorescence channel(exct:488nm).

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 discloses an antibody prodrug conjugate and preparation and application thereof. The antibody prodrug conjugate is formed by coupling an antibody and a prodrug molecule with a protectedfunctional group through a linker responding to a microenvironment, and has a structure shown in the description, wherein Ab is the antibody, L represents a connecting unit for connecting the antibodyand a drug molecule, D represents the drug molecule, X represents the protected functional group on the drug molecule, and Cage represents a protecting group. The antibody prodrug conjugate has enhanced tissue penetration and lower toxic and side effects, and can be used for treating tumors, immune diseases or infectious diseases. In particular, the antibody prodrug conjugate targeting the tumorsis stimulated by a tumor microenvironment to release the prodrug molecule, the prodrug molecule do not kill normal cells and are specifically activated after penetrating into tumor tissues, and compared with existing ADC drug molecules, the antibody prodrug conjugate improves tumor penetration, reduces the toxic and side effects, and has a good tumor treatment effect.

Description

technical field [0001] The invention relates to the field of biotechnology drugs, in particular to a preparation method and application of an antibody prodrug conjugate (ProADC). Background technique [0002] In the field of tumors, chemotherapy is still one of the most important treatment methods, but its development is greatly limited due to toxic and side effects. The emergence of antibody-drug conjugates (ADCs) provides a new solution to the above problems. By conjugating drug molecules and antibodies, people take advantage of the targeting of antibody molecules to deliver drug molecules around the tumor, thereby achieving tumor-specific killing. So far, 4 ADC drugs have been launched in the world, and more than 40 ADC drugs are in the clinical development stage. Overall, the development prospects of ADC drugs are broad. [0003] However, for ADC drugs, due to their large size and slow cell internalization rate, ADC drugs have poor tissue penetration ability, so they a...

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
IPC IPC(8): A61K47/68A61K47/54A61K31/704A61P35/00A61P37/02A61P31/00
Inventor 陈鹏林锋
Owner PEKING UNIV
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