Composite siRNA nanocarrier and preparation method and application thereof

A nanocarrier and autologous technology, applied in the field of biomedicine, can solve the problem that serum albumin cannot target tumor metastatic organs, and achieve the effects of excellent stability, high transfection, and good biocompatibility.

Active Publication Date: 2019-01-04
NANTONG UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] (4) Serum albumin alone cannot target tumor metastases

Method used

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  • Composite siRNA nanocarrier and preparation method and application thereof
  • Composite siRNA nanocarrier and preparation method and application thereof
  • Composite siRNA nanocarrier and preparation method and application thereof

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Experimental program
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Embodiment 1

[0034] A method for preparing a composite siRNA nanocarrier, comprising the following steps:

[0035] Step 1. Preparation of cationic albumin: Accurately weigh 100 mg of bovine serum albumin (BSA), completely dissolve it in 25 mL of ultrapure water, and adjust the rotating speed of magnetic stirring to avoid too fast generation of foam. After it is completely dissolved, add 25 mL of 1 mol / L ethylamine, mix well, adjust the reaction pH to 4.75 with dilute hydrochloric acid, then add 25 mg of carbodiimide hydrochloride (EDC·HCL) and slowly add to BSA In the mixed solution with ethylamine, add N-hydroxysulfosuccinimide (NHS) after 15min, the molar ratio of carbodiimide hydrochloride to N-hydroxysulfosuccinimide is 4:5,25 After reacting at ℃ for two hours, 0.55 mL of 4M acetic acid-sodium acetate buffer solution (PH=4.75) was added to terminate the reaction. Put the final reaction solution into a dialysis bag with a molecular weight cut-off of 8000-14000, and change the water eve...

Embodiment 2

[0041] A method for preparing a composite siRNA nanocarrier, comprising the following steps:

[0042] Step 1. Preparation of cationic albumin: Accurately weigh 100 mg of human serum albumin (HSA), completely dissolve it in 25 mL of ultrapure water, and adjust the speed of magnetic stirring to avoid foaming too quickly. After it is completely dissolved, add 25mL of 1mol / L ethylamine, mix well and adjust the reaction pH to 4.75 with dilute hydrochloric acid, then slowly add 25mg of dissolved carbodiimide hydrochloride (EDC·HCL) Into the mixed solution of BSA and ethylamine, add N-hydroxysulfosuccinimide (NHS) after 15 minutes, the molar ratio of carbodiimide hydrochloride to N-hydroxysulfosuccinimide is 4:5 , react at 37° C. for two hours, and add 0.55 mL of 4M acetic acid-sodium acetate buffer solution (PH=4.75) to terminate the reaction. Put the final reaction solution into a dialysis bag with a molecular weight cut-off of 8000-14000, and change the water every two hours. Af...

Embodiment 3

[0048] A method for preparing a composite siRNA nanocarrier, comprising the following steps:

[0049] Step 1. Preparation of cationic albumin: Accurately weigh 100 mg of bovine serum albumin (BSA), completely dissolve it in 25 mL of ultrapure water, and adjust the rotating speed of magnetic stirring to avoid too fast generation of foam. After it is completely dissolved, add 25mL of 1mol / L ethylamine, mix well and adjust the reaction pH to 4.75 with dilute hydrochloric acid, then slowly add 150mg of dissolved carbodiimide hydrochloride (EDC·HCL) Into the mixed solution of BSA and ethylamine, add N-hydroxysulfosuccinimide (NHS) after 15 minutes, the molar ratio of carbodiimide hydrochloride to N-hydroxysulfosuccinimide is 4:5 , reacted at 25° C. for two hours, and added 0.55 mL of 4M acetic acid-sodium acetate buffer solution (PH=4.75) to terminate the reaction. Put the final reaction solution into a dialysis bag with a molecular weight cut-off of 8000-14000, and change the wat...

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Abstract

The invention discloses a composite siRNA nanocarrier, comprising exosome lipid membrane from autologous tumor cells and siRNA-carrying cationic serum albumin covered in the exosome lipid membrane. The invention also discloses a preparation method and application of the composite siRNA nanocarrier. The composite siRNA nanocarrier has the advantages of high transfection, specific targeting propertyof cancer metastatic tissues, good biocompatibility, good stability, zero immunogenicity and the like.

Description

technical field [0001] The present invention relates to the field of biomedicine. More specifically, the present invention relates to a composite siRNA nanocarrier and its preparation method and application. Background technique [0002] RNA interference refers to post-transcriptional gene silencing due to target mRNA degradation or translational repression. siRNA can effectively and specifically block the expression of homologous genes in vivo through RNA interference, promote the degradation of homologous mRNA, induce cells to show a specific gene deletion phenotype and silence a certain oncogene, that is, RNAi method, which is effective in almost all tumors. It has been proved to have a good tumor suppressor effect in the model. However, naked siRNA is easily degraded by ribozyme (RNase) in vivo, has a short half-life, and low transfection efficiency. Therefore, how to efficiently introduce this specific silencing siRNA into target organs without damaging other normal t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/51A61K47/46A61K47/64A61K31/713A61K48/00A61P35/00
CPCA61K9/5068A61K31/713A61K48/005A61K47/643A61P35/00
Inventor 朱红艳赵刘婉韩伟莉
Owner NANTONG UNIVERSITY
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