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Nano hydroxyapatite-gene-medicament complex as well as preparation method and application thereof

A technology of nano-hydroxyapatite and drug complexes, applied in the field of biomedicine, can solve the problems of large cell toxicity and side effects, low efficiency of hydroxyapatite gene transfection, and patient harm, and achieve good biocompatibility, Favorable effect of endocytosis and good degradability

Active Publication Date: 2015-01-21
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, anti-tumor drugs themselves have defects: huge cytotoxicity and serious side effects, which will bring great harm to patients clinically
However, the existing hydroxyapatite gene transfection efficiency is not high, and it is particularly important to prepare a nano-hydroxyapatite with high transfection efficiency

Method used

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  • Nano hydroxyapatite-gene-medicament complex as well as preparation method and application thereof
  • Nano hydroxyapatite-gene-medicament complex as well as preparation method and application thereof
  • Nano hydroxyapatite-gene-medicament complex as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: Preparation and toxicity analysis of nano-hydroxyapatite

[0054] Calcium nitrate solution is used as calcium salt, and diammonium hydrogen phosphate solution is used as phosphate. Take 50mL of 6.25mmol / L calcium nitrate solution and cool it at 0°C; add 50mL of 3.74mmol / L diammonium hydrogen phosphate solution dropwise into the calcium carbonate solution, and maintain 200rpm / min; after the dropwise addition is completed, continue stirring for 60min ; Slowly add 200 μL of sodium bicarbonate solution, and continue stirring for 60 minutes; slowly adjust the pH of the reaction system to 10-13 with concentrated ammonia water, and continue stirring for 60 minutes after the adjustment is completed; slowly add 200 μL of polyethylene glycol-400 dropwise, and continue stirring for 60 minutes; Raise the temperature of the reaction device to 90°C, and continue stirring for 60 minutes; stop heating, and after the reaction system cools down to room temperature, wash the pr...

Embodiment 2

[0057] Example 2: Preparation of nano-hydroxyapatite-gene-drug complex

[0058] Add 60 μg of the anti-tumor gene p53 into 1 mL of nano-hydroxyapatite suspension, vortex, mix evenly, and let it stand for 30 minutes, then add 10 μg doxorubicin, vortex, mix evenly, let stand for 30 minutes, and set the temperature at 10000rpm / Centrifuge for 5 min, remove the supernatant, and then suspend and mix with 1 mL of deionized water to obtain a complex of nano-hydroxyapatite adsorbed genes and drugs.

Embodiment 3

[0059] Example 3: Determination of drug loading of nano-hydroxyapatite

[0060] The load of nano-hydroxyapatite anti-tumor gene p53 was determined by agarose gel electrophoresis. Add 2, 4, 6, 8, 10 μg p53 to 100 μL nano-hydroxyapatite, vortex, mix well, let stand for 30 minutes, take 20 μL dropwise into each agarose gel well, the voltage is 100V . Pictures were acquired using a gene gel imaging system. From Figure 6 It can be seen that there is a faint band in the 8 μL gel well, but no band in the 6 μL gel well, indicating that the drug loading of nano-hydroxyapatite against the tumor gene p53 is not less than 60 μg / mL.

[0061] The loading capacity of nano-hydroxyapatite antitumor drug doxorubicin was determined by centrifugation-measurement of absorbance value. Add 5, 10, 20, 30 and 40 μg of Dox to 100 μL of nano-hydroxyapatite-gene, vortex, mix evenly, let stand for 30 minutes, centrifuge at 10000 pm / min for 5 minutes, absorb the supernatant, and measure its Absorbanc...

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Abstract

The invention discloses a nano hydroxyapatite-gene-medicament complex as well as a preparation method and application thereof, belonging to the field of biomedicine. The complex disclosed by the invention comprises nano hydroxyapatite, anti-tumor genes and an anti-tumor medicament, wherein the size of nano hydroxyapatite is 20-300nm, the genes are anti-tumor genes and the medicament is an anti-tumor medicament. The preparation method comprises the following steps: mixing a calcium salt with a phosphate solution to prepare nano hydroxyapatite, adsorbing the anti-tumor genes to form a nano hydroxyapatite-gene precursor complex and further adsorbing the anti-tumor medicament by the precursor complex to form the complex, wherein the complex can efficiently pass through cancer cell membranes and promote the apoptosis of cancer cells. Hydroxyapatite with good biocompatibility is used as a co-transfer carrier, thereby having high safety, high loading rate of the genes and the medicament, high transfection efficiency and great effect of killing the cancer cells. Taking nano-hydroxyapatite as the co-transfer carrier of the genes and the medicament has great application prospects in the field of treatment of cancers.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to a nano-hydroxyapatite-gene-drug compound, a preparation method and an application. Background technique [0002] In recent years, the common methods for treating cancer are surgery, radiotherapy and chemotherapy, among which chemotherapy drugs such as doxorubicin, paclitaxel, cisplatin, etc. are being widely used. Taking doxorubicin as an example, as a spectrum non-specific cell cycle anticancer drug, doxorubicin can be embedded in RNA and DNA to inhibit cell cycle and promote tumor cell apoptosis. However, antitumor drugs have their own defects: huge cytotoxicity and serious side effects, which will bring great harm to patients in clinical practice. [0003] Gene therapy has been rapidly developed as a cure for genetically deficient diseases. Mutations of tumor suppressor genes have been found in most cancer cells, and the reintroduction of tumor suppressor gene...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K47/04A61K47/32A61K47/48A61K31/704A61P35/00
Inventor 孔祥东李泽豪陈岑叶婷蔡玉荣李麟涉赵瑞波
Owner ZHEJIANG SCI-TECH UNIV
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