Cyclodextrin soluble ACE2 as well as preparation method and application thereof

A technology of ACE2 and cyclodextrin, which is applied in the field of cyclodextrin-soluble ACE2 and its preparation, can solve the problems of affecting the atomization effect and poor water solubility of ACE2 protein

Inactive Publication Date: 2020-08-14
首都医科大学附属北京康复医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In order to solve the poor water solubility of ACE2 protein in the above-mentioned background technology, the present invention affects the technical problem of atomization effect when performing atomization treatment, and provides a kind of cyclodextrin-soluble ACE2 and its preparation method and application

Method used

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  • Cyclodextrin soluble ACE2 as well as preparation method and application thereof
  • Cyclodextrin soluble ACE2 as well as preparation method and application thereof
  • Cyclodextrin soluble ACE2 as well as preparation method and application thereof

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Embodiment 1

[0035] A preparation method of cyclodextrin-soluble ACE2, comprising the following steps:

[0036] S1. Dissolve ACE2 and hydroxypropyl-β-cyclodextrin in acetonitrile, then place the solution in a cellulose dialysis bag with a molecular weight cut-off of 500, and dialyze in water for 48 hours to obtain a dialysate containing insoluble matter. The ACE2 The molar ratio to hydroxypropyl-β-cyclodextrin is 1:10;

[0037] S2, the dialysate in step S1 is centrifuged, and the frequency of centrifugation is 1000r / min, and the time of centrifugation is 5min, gets the solution after centrifugation and carries out vacuum freeze-drying to obtain cyclodextrin-soluble ACE2.

[0038] The cDNA sequence of the ACE2: EEQAKTFLDKFNHEAEDLFYQSS-G-LGKGDFR;

[0039] The molecular structural formula of described cyclodextrin is shown in following formula (I):

[0040]

[0041] Specifically, in step S2, the specific operation of the vacuum freeze-drying is to pour the solution into the freeze-drying b...

Embodiment 2

[0044] The cyclodextrin-soluble ACE2 in Example 1 of the present invention can also be prepared by the following method:

[0045] 1. Gradual drop method: Dissolve ACE2 alone in an organic solvent such as acetonitrile, and then add it dropwise to the continuously stirring, uniformly dispersed hydroxypropyl-β-cyclodextrin solution. After vigorous stirring for 16h, it was filtered. The organic solvent was removed by dialysis, and the cyclodextrin-soluble ACE2 was obtained by freeze-drying.

[0046] 2. Grinding method: under normal temperature conditions, add ACE2 and hydroxypropyl-β-cyclodextrin into a mortar at a certain molar ratio, add a small amount of pure water to grind for 4 hours, then filter it, freeze-dry to obtain cyclodextrin Soluble ACE2.

[0047] 3. Ultrasonic method: put ACE2 and hydroxypropyl-β-cyclodextrin into a vial at a certain molar ratio, add an appropriate amount of deionized water, ultrasonicate for 10 minutes, filter, and freeze-dry to obtain cyclodextr...

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Abstract

The invention belongs to the field of prevention of coronavirus infection, and particularly relates to cyclodextrin soluble ACE2 as well as a preparation method and application thereof. The cyclodextrin soluble ACE2 comprises ACE2 and cyclodextrin, and the molar ratio of the cyclodextrin to the ACE2 is 1: (5-15). The preparation method comprises the following steps: the ACE2 and the cyclodextrin are dissolved in an organic solvent, then, the solution is placed in a cellulose dialysis bag with molecular weight cut-off of 100-1000, dialysis is performed in water for 40-50 h, an obtained dialysate containing insoluble matter is centrifuged, and the centrifuged solution is subjected to vacuum freeze-drying to obtain the cyclodextrin soluble ACE2. The cyclodextrin soluble ACE2 can solve the technical problem that existing ACE2 protein has poor water solubility and affects the atomization effect during atomization therapy.

Description

technical field [0001] The invention belongs to the field of preventing coronavirus infection, and in particular relates to a cyclodextrin-soluble ACE2 and its preparation method and application. Background technique [0002] Coronavirus is a relatively common human virus, causing 10% of common colds, but fewer of the more serious coronaviruses. Coronavirus has Spike protein (S-protein), which contains two subunits, S1 and S2. Spike protein is the most important pathogenic protein of coronavirus (Fig. 1), which helps the virus bind to the transmembrane receptor protein (ACE2) on the human cell membrane, thereby releasing the virus RNA into the cell. [0003] Studies have proved that SARS-CoV-2 has the same infection mechanism as SARS-CoV. It enters the cell through the combination of Spike protein and ACE2 protein. ACE2 protein may become a breakthrough in the study of new coronaviruses. ACE2 was identified as an essential receptor for SARS in vivo infection, and ACE2-Sp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/48A61K47/69A61P31/14
CPCA61K47/6951A61K38/4813C12Y304/17023A61P31/14
Inventor 席家宁叶霖王长琛郄淑燕刘宗建
Owner 首都医科大学附属北京康复医院
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