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Protein-based nanoparticles for delivery of multispecific antibodies and their applications and preparation methods

A multi-specific antibody and nanoparticle technology, applied in the field of medicine, to achieve the effect of enhanced therapeutic effect, excellent stability, and huge clinical application potential

Active Publication Date: 2022-04-19
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] How to realize a simple, convenient, stable and biocompatible antibody delivery system, and realize the efficient delivery of therapeutic antibodies in vivo under low-cost conditions, especially the efficient delivery of multispecific antibodies, still needs further research breakthroughs

Method used

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  • Protein-based nanoparticles for delivery of multispecific antibodies and their applications and preparation methods
  • Protein-based nanoparticles for delivery of multispecific antibodies and their applications and preparation methods
  • Protein-based nanoparticles for delivery of multispecific antibodies and their applications and preparation methods

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preparation example Construction

[0067] This embodiment also provides a method for preparing the above-mentioned protein-type nanoparticles, comprising the following steps:

[0068] (1) mixing the protein with water or an aqueous solution to obtain a water phase; mixing the polyester with an organic solvent to obtain an oil phase;

[0069] (2) preparing the water phase and the oil phase described in step (1) into an oil-in-water emulsion;

[0070] (3) Separating and purifying the emulsion to obtain protein nanoparticles.

[0071] In some of these embodiments, in step (1), the mass volume concentration of the protein in the aqueous phase is 5-20 mg / mL.

[0072] In some embodiments, in step (1), the concentration of the polyester in the oil phase is 1-10 mg / ml.

[0073] In some of these embodiments, in step (1), the organic solvent is at least one of chloroform, dichloromethane and ethyl acetate. More specifically, when the molecular weight of the polymer is small (7200-137000 Daltons), the mixed system of e...

Embodiment 1

[0127] Embodiment 1, preparation of albumin aliphatic polyester nanoparticles

[0128] Prepare a chloroform solution of ester-capped L-polylactide (PLLA-COOR-1100K) at a concentration of 5 mg / mL, dissolve 10 mg of human serum albumin in 1 mL of 0.9% sodium chloride aqueous solution, and prepare 1% (w / v) human serum albumin solution, take 200 μL of ester-capped L-polylactide chloroform solution in a 15mL centrifuge tube, add 1mL of 1% (w / v) human serum albumin solution, and use an ultrasonic cell disruptor Emulsification: Ultrasonic power is 130W, amplitude is 50%, ultrasonic 5s stop 2s, the total ultrasonic time is 2min, after the ultrasonic is finished, the emulsion is washed out of the centrifuge tube with ultrapure water, transferred to a 250mL round bottom flask, using a rotary evaporator, According to the order of vacuum degree of 300 / 120 / 70 / 40mbar, the chloroform was kept for 5 minutes to remove, and the albumin aliphatic polyester nanoparticles were collected after rot...

Embodiment 2

[0130] Embodiment 2, purification method of albumin aliphatic polyester nanoparticles

[0131] Method 1 (centrifugation method): Take the granule 1 obtained in Example 1, and use a desktop microrefrigerated centrifuge to centrifuge at low speed (3000rpm, 4°C) for 10min to remove unassembled water-insoluble polyester, and transfer the supernatant to a new The EP tube was centrifuged at high speed (15000rpm, 4°C) for 60 minutes to separate free albumin and nanoparticles, and the free albumin in the supernatant was removed, and the lower pellet was resuspended in 1×PBS for later use.

[0132] Method 2 (dialysis method): Take the granule 1 obtained in Example 1, and use a dialysis bag (Shanghai Green Bird) with a molecular weight cut-off of 14000 Da for dialysis. The dialysate is 1×PBS. Replace them all, and take the albumin aliphatic polyester nanoparticles in the dialysis bag for later use.

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Abstract

The invention relates to a protein-type nanoparticle for multispecific antibody delivery and its application and preparation method. The protein-type nanoparticles include polyester and a protein having a hydrophobic domain, and the hydrophobic domain of the protein is combined with the polyester through hydrophobic interactions; the protein is at least one of albumin, globulin and cell wall protein A sort of. The protein-type nanoparticles of the present invention have excellent stability and biocompatibility, and can prepare a multispecific antibody delivery platform (αFc-NP) by bonding anti-IgG-Fc antibodies or anti-IgG-Fc antibody fragments, αFc-NP It can stably, quickly and easily bind to multiple specific antibodies through antigen-antibody interaction and enhance the therapeutic effect of specific antibodies.

Description

technical field [0001] The invention relates to the field of medical technology, in particular to a protein-type nanoparticle used for multispecific antibody delivery and its application and preparation method. Background technique [0002] Monoclonal antibodies (mAbs) are the largest class of therapeutic proteins and can be used for a variety of diseases, such as tumors, inflammation, and autoimmune diseases. Most therapeutic antibodies achieve therapeutic purposes by blocking receptors or ligands, thereby reducing the activity of certain signaling pathways. Since the first monoclonal antibody used to block CD3 in the treatment of renal transplant rejection was approved by the FDA in 1986, after more than 30 years of rapid development, monoclonal antibody drugs include therapeutic antibodies and antibody derivatives (including antigen-binding fragment Fab , Fc fusion proteins, etc.), has become the fastest growing segment of biopharmaceuticals in the world, and several "su...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/62A61K47/59A61K47/69A61K39/395A61P37/00
CPCA61K47/62A61K47/593A61K47/6921A61K39/395A61P37/00A61K47/6937A61K47/6929
Inventor 王均曹志婷沈松叶倩妮都小姣杨显珠袁友永
Owner SOUTH CHINA UNIV OF TECH
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