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Method for preparing stable albumin nano-particles by virtue of thermal denaturation

A technology of nano-albumin and albumin, which is applied in the direction of pharmaceutical formulations, organic active ingredients, and medical preparations containing active ingredients, etc. It can solve the problems of unfavorable production quality stability, and the inability of nanoparticles to withstand the dilution of buffer solutions and serum albumin solutions , endangering the life safety of patients, etc.

Inactive Publication Date: 2015-04-08
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the albumin nanoparticles prepared by the Nab method (patent 200310123461.X) also have some shortcomings: the nanoparticles prepared by this method cannot withstand the dilution of buffer solution and serum albumin solution; the long time of high-pressure homogenization will affect the stability of the drug Sexuality, which can cause drug degradation; drug nanocrystals will aggregate into larger particles within 24 hours, which is not conducive to the production of stable quality nanoparticle preparations
[0004] The preparation methods of albumin nanoparticles also include anti-solvent method, emulsification solidification method, disulfide bond breaking method (CN102048695A), etc., in these preparations, organic solvents such as ethanol, acetone, methylene chloride, chloroform, glutaraldehyde, mercaptoethanol, etc. Or toxic reagents are indispensable, and the use of these organic solvents or toxic reagents may also become a hidden danger that endangers the life safety of patients

Method used

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  • Method for preparing stable albumin nano-particles by virtue of thermal denaturation
  • Method for preparing stable albumin nano-particles by virtue of thermal denaturation
  • Method for preparing stable albumin nano-particles by virtue of thermal denaturation

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Example 1, preparation of stable bovine serum albumin nanoparticles.

[0036] Using bovine serum albumin and vanillin as raw materials, bovine serum albumin nanoparticles stabilized by disulfide bonds and amide bonds were prepared.

[0037] The preparation method is as follows:

[0038] Dissolve 250 mg of bovine serum albumin and 14.4 mg of vanillin in 50 mL of purified water, heat at 70 °C for 2 h and then cool to room temperature, then place the prepared nanoparticle solution in a dialysis bag and dialyze in purified water at 4 °C for 24 h. That is, stable bovine serum albumin nanoparticles are obtained, and the average particle diameter of the albumin nanoparticles is 50-300 nm.

[0039] The stepwise degradation process of the prepared nanoparticles in trypsin solution can be obtained from figure 1 The particle size change curve of a and figure 1 It is confirmed in the SDS-PAGE gel electrophoresis of b.

[0040] The stepwise degradation process of the prepared nano...

Embodiment 2

[0041] Example 2, preparing stable bovine serum albumin nanoparticles.

[0042] Using bovine serum albumin and vanillin as raw materials, bovine serum albumin nanoparticles stabilized by disulfide bonds and amide bonds were prepared.

[0043] The preparation method is as follows:

[0044] Dissolve 250 mg of bovine serum albumin and 14.4 mg of vanillin in 50 mL of purified water, heat at 80°C for 1 hour and cool to room temperature, then place the prepared nanoparticle solution in a dialysis bag and dialyze in purified water at 4°C for 24 hours. That is, stable bovine serum albumin nanoparticles are obtained, and the average particle diameter of the albumin nanoparticles is 50-500 nm.

[0045] The stability of the resulting albumin nanoparticles stored in phosphate buffered saline at 4°C can be obtained from Figure 4 look out.

[0046] The change of the free sulfhydryl groups of the obtained albumin nanoparticles during the preparation process can be obtained from Figure ...

Embodiment 3

[0049] Example 3, preparing a stable doxorubicin hydrochloride-human serum albumin nanoparticle drug delivery system.

[0050] Using human serum albumin, doxorubicin hydrochloride and vanillin as raw materials, a drug delivery system of human serum albumin nanoparticles loaded with doxorubicin hydrochloride stabilized by disulfide bonds and amide bonds was prepared.

[0051] The preparation method is as follows:

[0052] Dissolve 250 mg of human serum albumin and 14.4 mg of vanillin in 50 mL of purified water, and heat at a constant temperature of 70° C. for 2 hours to obtain a human serum albumin nanoparticle solution. After the above-mentioned human serum albumin nanoparticle solution was cooled to room temperature, 2.5 mL of a 5 mg / mL doxorubicin hydrochloride solution was heated therein, and placed overnight in the dark. The obtained nano-suspension of human serum albumin was placed in a dialysis bag, and dialyzed in purified water at 4° C. for 24 hours to obtain human se...

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Abstract

The invention belongs to the field of preparation of biological medical materials, and relates to a method for preparing stable albumin nano-particles by virtue of thermal denaturation. The method comprises the following steps: (1) adding vanillic aldehyde or an analogue thereof to form intermolecular disulfide bonds by virtue of inter-reaction of free sulfhydryl groups on albumin molecules under a heating condition; (2) enabling amino groups inside and among molecules to react with carboxyl so as to form amido bonds; and (3) enabling amino groups on the albumin molecules to react with aldehyde groups on vanillic aldehyde or the analogue thereof to form chemical bonds of Schiff base and the like so as to form stable nano-particles in an aqueous solution. Any organic solvent is not introduced during preparation, so that the prepared nano-particles are safe and nontoxic, and can well entrap antitumor drugs including paclitaxel, doxorubicin hydrochloride and the like. Moreover, the carrier has an oxidation reduction response in a tumor cell internal environment, and can open disulfide bonds to release drugs under the action of reducing glutathione in cells. The method provided by the invention is simple in process, convenient to operate and suitable for industrial mass production.

Description

technical field [0001] The invention relates to a method for thermally denaturing and preparing stable albumin nanoparticles, which belongs to the technical field of biomedical material preparation. Background technique [0002] Chemotherapy is currently the most commonly used tumor treatment strategy in clinical practice. However, due to the lack of selectivity of anticancer drugs, it has strong toxic and side effects on normal cells and tissues. paclitaxel injection Clinically, there is a strong allergic reaction, and doxorubicin hydrochloride injection has strong cardiotoxicity. Selecting an excellent drug delivery carrier can realize the delivery of the drug to the target site, reduce the toxic and side effects of anti-tumor drugs, and improve the chemotherapy effect. [0003] Albumin is the protein with the most content in plasma. It has the advantages of non-toxic, safe, biodegradable, and non-immunogenic. It has been clinically used as a drug delivery carrier and i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/51A61K47/42A61K31/704A61K31/337A61P35/00
Inventor 李芳辛俊勃刘建平朱家壁施秦
Owner CHINA PHARM UNIV
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