Fusion protein and encoding gene and preparation method of fusion protein as well as pharmaceutical composition and preparation method of pharmaceutical composition

A technology of fusion protein and coding gene, which is applied in the field of pharmaceutical composition and its preparation, can solve the problems of unstable drug release rate, poor drug loading stability, difficulty in carrying hydrophobic drugs, etc., and achieve good biocompatibility , High drug loading efficiency, good drug release effect

Active Publication Date: 2014-06-25
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the existing biomaterial carriers are difficult to carry hydrophobic drugs, and even if they can be loaded with hydrophobic drugs, it is difficult to control the release of the hydrophobic drugs carried by them at the target site, and even the stability of the drug loading is not good, and the drug release is difficult. Speed ​​instability and other defects

Method used

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  • Fusion protein and encoding gene and preparation method of fusion protein as well as pharmaceutical composition and preparation method of pharmaceutical composition
  • Fusion protein and encoding gene and preparation method of fusion protein as well as pharmaceutical composition and preparation method of pharmaceutical composition
  • Fusion protein and encoding gene and preparation method of fusion protein as well as pharmaceutical composition and preparation method of pharmaceutical composition

Examples

Experimental program
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preparation example Construction

[0030] The present invention also provides a method for preparing a fusion protein, which comprises expressing the coding gene of the present invention in a strain to obtain the fusion protein.

[0031] In the present invention, there are many methods for expressing the coding gene in the strain, which can be various methods conventionally used in the field. For example, the literature (Setting the chaperonin timer: A two-stroke, two-speed , protein machine. Grason JP, Gresham JS, Lorimer GH, etc., Proceedings of the National Academy of Sciences, 2008, 105(45): 17339-17344).

[0032] In the preparation method of the fusion protein of the present invention, the nucleotide sequence of the gene encoding iRGD peptide and the method of introducing the nucleotide sequence of the gene encoding the molecular chaperone GroEL into the strain can be conventionally used in the field Various methods, the invention of the present invention does not lie in this. Those skilled in the art know from...

Embodiment approach

[0049] According to a preferred embodiment of the present invention, the method for preparing the pharmaceutical composition includes contacting the pharmaceutical compound with the fusion protein of the present invention in the presence of a solvent, and the amount of the pharmaceutical compound is The amount is 0.005-0.1 parts by weight, and the contact conditions include a pH of 6-8, a temperature of 4-50°C, and a time of 12-48 hours. The materials obtained after the contact are then centrifuged, ultrafiltered and washed in sequence.

[0050] The invention also provides a pharmaceutical composition prepared by the above method of the invention.

[0051] Hereinafter, the present invention will be explained in further detail through examples. Among them, unless otherwise specified, the reagents used are all commercially available products.

[0052] In the following examples of the present invention, the used doxorubicin hydrochloride CAS number: 25316-40-9, purchased from sigma;

...

preparation example 1

[0059] This preparation example is used to obtain the iGroEL fusion protein provided by the present invention.

[0060] The amino acid sequence of the iRGD peptide and the amino acid sequence GGG connected by the bridge were inversely deduced into the gene sequence according to the genetic code to obtain the nucleotide sequence of the cDNA sequence of the iRGD peptide and the cDNA sequence of GGG, and the nucleotide sequence was combined with GroEL The primer pair (for example, the forward primer HS43 shown in SEQ ID No: 4 and the reverse primer HS45 shown in SEQ ID No: 5) are connected to obtain the amplification primer, and the amplification primer is used according to the method on molecular cloning The cDNA sequence of GroEL was amplified to obtain the recombinant DNA sequence of iGroEL fusion protein. Among them, the gene containing the iRGD peptide and the bridge-connected gene are connected with the 3'-end of the gene of the molecular chaperone GroEL sequence following PCR...

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Abstract

The invention discloses a fusion protein, the amino acid sequence of which contains an iRGD peptide sequence shown as SEQ ID No:1 and a molecular chaperone GroEL sequence shown as SEQ ID No:2. The invention further discloses an encoding gene of the fusion protein, the sequence of which is a nucleotide sequence capable of encoding the fusion protein. The invention further discloses a preparation method of the fusion protein. The preparation method comprises the following step of expressing the encoding gene in a bacterial strain to obtain the fusion protein. A method of preparing a pharmaceutical composition comprises the following steps of contacting the pharmaceutical compound with the fusion protein disclosed by the invention to obtain a contacted material in the presence of a solvent. The invention further discloses the pharmaceutical composition prepared by the method of preparing the pharmaceutical composition. The fusion protein provided by the invention can stably load a hydrophobic drug, and is high in drug-carrying efficiency and good in drug release effect. The pharmaceutical composition prepared by the fusion protein provided by the invention has the advantages of good biocompatibility and good drug release effect.

Description

Technical field [0001] The present invention relates to the field of biological materials, in particular, to a fusion protein, a fusion protein encoding gene, a preparation method of a fusion protein, a pharmaceutical composition and a preparation method thereof. Background technique [0002] In recent years, the development and research of biomaterial drug carriers has received extensive attention. Biomaterials drug carriers have many advantages, such as: good biosafety and can be modified by genetic engineering. [0003] Because protein biomaterials are biologically active substances related to various cell functions in the organism, they have good biocompatibility and controllable biodegradability. The degraded short peptides and amino acids can also be absorbed by the human body. Therefore, compared with Other drug carrier systems and protein biomaterials have broader application prospects. [0004] However, the existing biomaterial carriers are difficult to carry hydrophobic d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62A61K47/42
Inventor 聂广军袁嫕孙翠骥
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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