Application of anti-BASIGIN humanized antibody in preparation of drugs for treating COVID-19

A technology of humanized antibody and coronavirus, applied in the direction of antiviral agents, antibodies, drug combinations, etc., can solve the problems of blocking antibodies, allergic and toxic reactions, etc., and achieve the effect of inhibiting chemotaxis

Pending Publication Date: 2020-07-17
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the repeated injection of mouse McAb into the human body will cause the patient to induce human anti mouse anti

Method used

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  • Application of anti-BASIGIN humanized antibody in preparation of drugs for treating COVID-19
  • Application of anti-BASIGIN humanized antibody in preparation of drugs for treating COVID-19
  • Application of anti-BASIGIN humanized antibody in preparation of drugs for treating COVID-19

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: The SPR method measures the interaction between the new coronavirus S protein (COVID-19Spike-RBD) and CD147

[0054] Based on surface plasmon resonance (SPR) technology, BIAcore 3000 system and CM5 sensor chip were used to measure the molecular binding kinetic parameters between the new coronavirus S protein (COVID-19Spike-RBD) and CD147. The detection buffer system is PBS (pH7.4), and the detection temperature is 25°C; 35 μg CD147 is immobilized on the surface of the CM5 chip by amino coupling; the interaction is detected by Kinetic Analysis / Concentration Series / Direct Binding mode, and the flow rate is set to 10 μL / min , the binding time is 10min, and the dissociation time is 10min; 0.4, 0.8, 1.0 and 1.2μM of the new coronavirus S protein (COVID-19Spike-RBD) of the samples to be tested are injected into different channels respectively. The results were fitted by BIAevaluation software to determine the kinetic constants.

[0055] The result is as figure 1S...

Embodiment 2

[0056] Example 2: The SPR method measures the interaction between the N protein of the new coronavirus and cyclophilin A (CyPA)

[0057] Based on surface plasmon resonance (SPR) technology, BIAcore 3000 system and CM5 sensor chip were used to determine the molecular binding kinetic parameters between the new coronavirus N protein and cyclophilin A (CyPA). The detection buffer system is PBS (pH7.4), and the detection temperature is 25°C; CyPA is immobilized on the surface of the CM5 chip by amino coupling; the chip is inactivated with 1M ethanolamine-HCl (Bio-Rad); Buffer (PBS / 0.005% Tween 20) was washed until the baseline stabilized. Inject 0.2, 0.4, and 0.8 μM of the new coronavirus N protein on the horizontal channel respectively, set the flow rate to 10 μL / min, the binding time to 10 minutes, and the dissociation time to 10 minutes; use the Kinetic Analysis / Concentration Series / Direct Binding mode to detect the interaction. The results were fitted by BIAevaluation software...

Embodiment 3

[0059] Embodiment three: SPR method measures the interaction between CD147 and cyclophilin A (CyPA)

[0060] SPR was used to monitor the binding of CD147 to cyclophilin A (CyPA) in real time, and the affinity between CD147 and CyPA was reflected by measuring the affinity constant (KD). Affinity determination of CD147 and cyclophilin A (CyPA) was performed with ProteOn XPR36 (Bio-Rad, XPR36) instrument. A GLC chip (Bio-Rad, 1765011) was activated with 0.04M EDC+0.01M sulfo-NHS (Bio-Rad). Dilute CD147 to 10mM with 10mM NaAc (pH 4.5), and inject it onto the chip at a speed of 30ul / min to couple the antigen to the activated chip through amino groups. Finally, the chip was inactivated with 1M ethanolamine-HCl (Bio-Rad); the chip was rotated 90 degrees and washed with buffer solution (PBS / 0.005% Tween 20) until the baseline was stable. 1 and 30 μM cyclophilin A (CyPA) samples were injected on the horizontal channel, respectively. The injection speed is 30ul / min. The sample bindi...

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Abstract

The invention discloses an application of an anti-human BASIGIN humanized antibody in preparation of drugs for treating COVID-19. The amino acid sequence of a light chain variable region of the anti-human BASIGIN humanized antibody is shown as SEQ ID NO:1, and the amino acid sequence of a heavy chain is shown as SEQ ID NO:3; and the nucleotide sequence of the light chain variable region is shown as SEQ ID NO:2, and the nucleotide sequence of the heavy chain variable region is shown as SEQ ID NO:4. The antibody can specifically recognize and bind a virus invasion host epithelial cell co-receptor CD147, block the interaction of the CD147 and COVID-19 S protein, and interaction of the CD147 and cyclophilin A (CyPA), thereby inhibiting the COVID-19 from infecting host cells.

Description

technical field [0001] The invention belongs to the technical field of novel antibody drugs in the treatment of viral pneumonia. Specifically, the invention relates to the application of an anti-BASIGIN humanized antibody for preparing a drug for treating novel coronavirus (COVID-19) pneumonia. Background technique [0002] Coronaviruses are a class of single-stranded positive-sense RNA viruses that can infect humans and various vertebrates. They are important pathogens that cause common colds and upper respiratory tract infections in humans, and pose a serious threat to human health. The novel coronavirus (COVID-19) that spread at the end of 2019 has the characteristics of rapid spread, severe epidemic and rapid progress. Caused progressive impairment of respiratory function and death in some patients. The current treatment of 2019-nCoV is mainly to support symptomatic treatment and antiviral treatment. There is a lack of specific drugs, and it is urgent to develop new pre...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61P31/14A61P11/00
CPCC07K16/2803A61P31/14A61P11/00A61K2039/505C07K2317/24
Inventor 陈志南朱平边惠洁张征尉丁杨向民
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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