Hepatitis B virus-like particle nano-carrier and drug delivery system

A hepatitis B virus, nano-carrier technology, applied in nano-drugs, viruses/phages, viruses, etc., to achieve the effects of good stability, high encapsulation stability, and not easy to leak

Pending Publication Date: 2022-06-10
河南省生物工程技术研究中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The encapsulation stability of targeted nanocarriers for drugs disclosed in this patent document CN109529044A is relatively poor under conditions close to physiological pH, and needs to be further improved

Method used

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  • Hepatitis B virus-like particle nano-carrier and drug delivery system
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  • Hepatitis B virus-like particle nano-carrier and drug delivery system

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1 Preparation of the hepatitis B virus-like particle nanocarrier of the present invention

[0047] Include the following steps:

[0048] (1) According to the amino acid sequence of the hepatitis B virus-like particle nanocarrier of the present invention, the nucleotide sequence encoding the hepatitis B virus-like particle nanocarrier is obtained by gene synthesis, and the expression vector plasmid pEt43 is constructed by Xho I / Nde I restriction site .1(a)—insert the RGD sequence between amino acids 78 and 82 of the HBc-170 protein (delete amino acids 79-81), and insert a connecting peptide between amino acid 78 and the RGD sequence GTSGTSGSSGSGGT, insert connecting peptide GGSGSSGSTG between RGD and the 82nd amino acid, obtain the amino acid sequence of RGD-HBc-170 protein (as shown in SEQ ID No.1); Add poly Histidine polypeptide (HHHHHH); according to the above description, synthesize the nucleotide sequence encoding the hepatitis B virus-like particle nano...

Embodiment 2

[0056] Embodiment 2 builds the drug delivery system of the present invention

[0057] 2.1 Drug loading

[0058] Take 100mL of the target protein (0.5mg / mL) and incubate with 50mL of pre-prepared dissociation solution at 25°C for 2h, the total volume of the solution is 150mL. At this point, 37.5 mg of doxorubicin was added to the above mixed solution, and the resulting mixed solution was incubated at 4° C. for 30 min with shaking. Afterwards, it was moved to a dialysis bag with a molecular weight of 3500 Da, first placed in 10% polymer solution (1 L) and dialyzed overnight at 4°C, and then replaced with 10% polymer solution after 12h and dialyzed at 4°C, After 2 hours, change to 0% polymer solution and continue dialysis at 4° C., during which the polymer solution is replaced 5 times until the dialysate is clear and transparent, and the dialysis can be stopped when the dialysate no longer turns red, that is, the encapsulation complex (delivery agent of the present invention) is...

Embodiment 3

[0073] Embodiment 3: Verify the purity of complex (drug delivery system of the present invention) by means of HPLC

[0074] (1) The detection wavelength of the detector of the high performance liquid chromatograph is 280nm (hepatitis B virus-like particle nanocarrier absorption peak of the present invention), 480nm (DOX absorption peak);

[0075] (2) the chromatographic column is an Ultrahydrogel 2000 chromatographic column;

[0076] (3) The column length of the chromatographic column is 300mm

[0077] (4) The inner diameter of the chromatographic column is 7.8mm

[0078] (5) The column temperature of the chromatographic column is 25°C

[0079] (6) The mobile phase of the chromatographic column is: 20mmol / L PBS, 150mmol / L NaCl;

[0080] (7) The chromatographic column flow rate: 0.6mL / min;

[0081] (8) By detecting the ultraviolet absorption value of the complex at 280nm and 480nm, it can be seen that there is a single absorption peak at about 14min, without other miscellan...

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Abstract

The invention belongs to the technical field of drug nano-carriers, and particularly relates to a hepatitis B virus-like particle nano-carrier and a drug delivery system. The vector has an amino acid sequence as follows: 79-81st amino acids of hepatitis B core protein HBc-170 are replaced with RGD sequences, and two ends of the RGD are respectively connected with 78th and 82nd amino acids of the HBc-170 by a connecting peptide to construct the vector. After the medicine is encapsulated by the hepatitis B virus-like particle nano-carrier, the encapsulation stability is obviously improved.

Description

technical field [0001] The invention belongs to the technical field of drug nanocarriers, and in particular relates to a hepatitis B virus-like particle nanocarrier and a drug delivery system. Background technique [0002] In the current clinical medicine, nanotechnology has made considerable progress, and is increasingly being used in many fields such as medicine, material science, biology, etc., and plays a pivotal role in various fields. The application of nanotechnology reflects the popularization of a new disease treatment mode, and has achieved good results in many academic research and clinical practice. The method of using nanoparticles as carriers to deliver tumor therapeutic drugs overcomes the disadvantages of traditional chemotherapy drugs, and provides the possibility to improve the efficacy of chemotherapy drugs and reduce their side effects. [0003] Among the various types of VLPs currently existing, virus-like particles based on hepatitis B virus core prote...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/005A61K47/42A61K47/64A61K9/51A61K31/704A61K45/00A61P35/00B82Y5/00B82Y30/00B82Y40/00
CPCC07K14/005A61K9/5169A61K47/645A61K45/00A61K31/704A61P35/00B82Y5/00B82Y30/00B82Y40/00C12N2730/10123C12N2730/10151C12N2730/10142
Inventor 王云龙李玉林王继创张怡青王敏王运从程蕾韩笑
Owner 河南省生物工程技术研究中心
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