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A kind of genetically engineered polypeptide nano hydrogel loaded with hydrophobic drugs and hydrophilic drugs and its preparation method

A nano-hydrogel, hydrophilic drug technology, applied in the field of genetically engineered polypeptide nano-hydrogel and its preparation, can solve the problems of tumor drug resistance, tissue and organ damage, lack of targeting, etc. Drug properties, long residence time, and the effect of improving loading rate and encapsulation rate

Inactive Publication Date: 2021-04-06
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But regardless of which drug, long-term monotherapy can lead to drug resistance in tumors
At the same time, due to the lack of targeting, only a small part of the directly injected medicinal solution reaches the tumor site, causing serious damage to other normal tissues and organs

Method used

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  • A kind of genetically engineered polypeptide nano hydrogel loaded with hydrophobic drugs and hydrophilic drugs and its preparation method
  • A kind of genetically engineered polypeptide nano hydrogel loaded with hydrophobic drugs and hydrophilic drugs and its preparation method
  • A kind of genetically engineered polypeptide nano hydrogel loaded with hydrophobic drugs and hydrophilic drugs and its preparation method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] PC 10 Preparation of ARGD: Use restriction endonucleases BamHI, NheI and SpeI to cut out the PC containing the target gene fragment 10 A and RGD fragments, and then use T4 DNA ligase to recombine the target gene fragments to obtain PC 10 The ARGD fragment is introduced into Escherichia coli for expression, then denatured and lysed with urea, centrifuged to obtain the protein, purified with a nickel column, dialyzed and freeze-dried to obtain the spongy protein.

[0031] to PC 10 The biosafety of ARGD is verified: PCs with different weights are weighed separately 10 The ARGD polypeptide is dissolved in 1 milliliter of cell culture medium DMEM, and the pH of the solution is adjusted to 7-8 to obtain cell culture fluids containing nano hydrogels with different concentrations. After incubating Hela with the hydrogel at different concentrations for 24 hours, the cell survival rate was examined by MTT method, and the results were as follows: figure 1 As shown: the cells s...

Embodiment 2

[0034] PC 10 Application of ARGD / PTX / DOX·HCL nanohydrogels: PC 10 ARGD polypeptide, DOX, PC 10 ARGD polypeptide / DOX, PC 10 ARGD polypeptide / PTX / DOX·HCL was dissolved in DMMEM cell culture medium and co-incubated with Hela cells for 4 hours, the results were as follows figure 2 Shown: It can be seen under the confocal microscope that individual DOX enters the cytoplasm and nucleus through molecular diffusion, while PC 10 ARGD polypeptide / DOX·HCL nano hydrogel and PC 10 ARGD polypeptide / PTX / DOX·HCL nanohydrogels only entered the cytoplasm, indicating that PC 10 ARGD polypeptide / DOX·HCL nano hydrogel and PC 10 ARGD polypeptide / PTX / DOX·HCL nano hydrogel entered Hela cells through phagocytosis.

Embodiment 3

[0036] PC 10 Slow drug release from ARGD / PTX / DOX·HCL nanohydrogels: mixing PC 10ARGD / PTX / DOX·HCL nanohydrogels were respectively placed in buffers of different pH (pH 7.4 PBS phosphate buffer, pH 5.0 acetate buffer). The release behaviors of DOX·HCL and PTX in different pH buffers were detected by UV absorption and HPLC analysis. It can be seen from the experimental results that the release of DOX·HCL and PTX in the acetate buffer at pH 5.0 is faster than in the neutral pH 7.4 PBS phosphate buffer, and the tumor environment is also acidic, which is conducive to the release of DOX·HCL and PTX in acidic tumors. The release in the environment, and the drug release time lasts for more than 48 hours, achieving an obvious slow release effect.

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Abstract

The invention belongs to the technical field of nano-biomedicine, in particular to a genetically engineered polypeptide nano-hydrogel that simultaneously carries hydrophobic drugs and hydrophilic drugs and a preparation method thereof. The nanohydrogels are made of PC 10 ARGD polypeptide hydrogel was simultaneously loaded with paclitaxel and doxorubicin hydrochloride to prepare nano-hydrogel particles. The present invention obtains PC by gene recombination method 10 ARGD peptide nanohydrogels using PC 10 ARGD polypeptide nano-hydrogels are loaded with hydrophobic drug paclitaxel and hydrophilic drug doxorubicin hydrochloride at the same time. Nano-hydrogel particles loaded with paclitaxel and doxorubicin hydrochloride not only have drug sustained-release and targeting effects, but also drug loading. The nano-hydrogel particles have a longer retention time in the tumor, which is conducive to the synergistic effect of paclitaxel and doxorubicin hydrochloride to achieve the efficacy of 1+1>2. Compared with single administration, the efficacy of paclitaxel and doxorubicin hydrochloride Combination administration can effectively avoid the occurrence of drug resistance.

Description

technical field [0001] The invention belongs to the technical field of nano-biomedicine, and in particular relates to a genetically engineered polypeptide nano hydrogel loaded with hydrophobic drugs and hydrophilic drugs and a preparation method thereof. Background technique [0002] A large number of studies have shown that doxorubicin can inhibit the synthesis of DNA and RNA, and has a killing effect on tumor cells in various growth cycles. PTX (paclitaxel) can promote microtubule polymerization and stabilize the polymerized microtubules, blocking the division of tumor cells. But no matter what kind of drug, long-term monotherapy will lead to drug resistance of tumors. At the same time, due to the lack of targeting, only a small part of the directly injected medicinal solution reaches the tumor site, causing serious damage to other normal tissues and organs. At present, many studies have explored that it can be used as a carrier of tumor drugs to reduce the occurrence of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/06A61K31/337A61K31/704A61K47/42A61P35/00
CPCA61K9/06A61K31/337A61K31/704A61K47/42A61P35/00A61K2300/00
Inventor 刘波金瑞美赵元弟
Owner HUAZHONG UNIV OF SCI & TECH
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