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Application of Shikonin and Its Derivatives as Gene Therapy Sensitizers

A gene therapy and derivative technology, applied in the field of gene therapy, can solve the problem of undetermined target liver cancer, and achieve the effect of strong anti-tumor effect

Inactive Publication Date: 2017-01-04
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies using rAAV as a carrier have achieved a certain inhibitory effect on liver cancer, but the best rAAV serotype targeting liver cancer has not been determined; as a lethal therapy, the unique targeting of liver cancer cells is crucial
In addition, in the clinical use of rAAV, it is very important to reduce the dosage and ensure safety. How to maximize the efficiency of rAAV infection with the best targeting of liver cancer, no one has proposed a solution so far

Method used

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  • Application of Shikonin and Its Derivatives as Gene Therapy Sensitizers
  • Application of Shikonin and Its Derivatives as Gene Therapy Sensitizers
  • Application of Shikonin and Its Derivatives as Gene Therapy Sensitizers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Effect of shikonin on rAAV infection efficiency

[0037] 1. Experimental materials and methods

[0038] 1. Compounds, Plasmids, Primers

[0039] High-performance liquid chromatography (HPLC) grade tripterine, pristimerin, shikonin, curcumin, apigenin, chrysin, luteolin, and salvianolic acid B salt were purchased from Sigma-Aldrich Co. LLC, St. Louis, USA, dissolved in DMSO at a concentration of 5 mg / ml as a stock solution. HPLC grade MG132 was purchased from Calbiochem, Merck KGaA, Darmstadt, Germany, 5 mg / ml dissolved in DMSO. HPLC grade bortezomib was purchased from Santa Cruz Biotechnology, Inc. Santa Cruz, USA, dissolved in DMSO at 5 mg / ml.

[0040] The pHelper plasmid was purchased from Agilent Technologies, Santa Clara, USA. pdsAAV-CBAp-EGFP and pAAV-CBAp-Fluc are prepared as follows: using molecular cloning technology, chicken actin promoter (CBAp) driven enhanced green fluorescent protein gene (EGFP) or luciferase gene (Fluc) Embedding into a pla...

Embodiment 2

[0078] Example 2 The mechanism of shikonin enhancing rAAV infection efficiency

[0079] 1. The mechanism of shikonin enhancing rAAV infection efficiency in vitro

[0080] Firstly, the coat protein of rAAV2-CB-Fluc was designed to be labeled with Cy3 dye, and then the transport of the labeled rAAV2-CB-Fluc-Cy3 virus coat in HeLa cells was observed with a Leica TCS SP5 confocal microscope. Before the official observation, the biological activity of the labeled rAAV2-CB-Fluc-Cy3 virus was detected, and HeLa cells were infected. The reporter gene expression in the high-dose group and the low-dose group could be detected, but it was higher than that of the unlabeled virus at the same dose. group significantly decreased [ Figure 5 .A]; In addition, the effect of shikonin on the infection efficiency of rAAV2-CB-Fluc-Cy3 was detected, and the results showed that shikonin could still significantly enhance its infection efficiency[ Figure 5 .B], indicating that Cy3 dye labeling has ...

Embodiment 3

[0085] Example 3 Shikonin Combined with rAAV3 Carrying Killer Genes in the Treatment of Liver Cancer

[0086] Trichosanthin (TCS) is derived from Radicestrichosanthis, a traditional Chinese medicine. Trichosanthin is a perennial climbing herb belonging to Cucurbitaceae Trichosanthes, Trichosanthis ( Trichosanthes kirilowii Maxim) or Bilateral Trichosanthes ( Trichosanthes rosthornii Herms) roots. The effects of trichosanthes are: clearing heat and purging fire, promoting body fluid and quenching thirst, reducing swelling and draining pus. It is mainly used to cure fever, thirst, thirst, jaundice, dryness of the lung and hemoptysis, carbuncle, hemorrhoids, etc. Consisting of 247 amino acid residues, TCS belongs to type I ribosome-inactivating proteins (Ribosome-inactivating proteins, RIP), and has various pharmacological activities such as inhibiting the growth of various tumor cells, anti-virus, immune regulation, and anti-AIDS virus. Shaw PC et al first confirmed the s...

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Abstract

The invention relates to the application of shikonin and its derivatives in the preparation of gene therapy sensitizers. The present invention has the advantages that: shikonin and its derivatives have relatively low toxicity as gene therapy sensitizers; when using shikonin and its derivatives in combination with gene therapy for malignant tumor treatment, shikonin and its derivatives The anti-cancer effect of the drug is superimposed with the enhanced effect of gene therapy, which can maximize the anti-tumor effect.

Description

technical field [0001] The invention relates to the field of gene therapy, specifically, the application of shikonin and its derivatives as gene therapy sensitizers. Background technique [0002] Lithospermum erythrorhizon (English name: Lithospermum erythrorhizon) is a traditional Chinese medicine commonly used in traditional Chinese medicine. Cold in nature, sweet and salty; Guixin, Liver Meridian; Efficacy: Cooling blood to stop bleeding, clearing away heat and detoxification; Indications: Congestion of blood heat, purple and black macule, opaque measles, sores, eczema, water and fire burns. Shikonin is one of the main active ingredients of Zicao, molecular formula: C 16 h 16 o 5 , molecular weight: 288.30 g / mol. Shikonin has anti-inflammatory effects by inhibiting tumor necrosis factor-a (TNF-a), which is an important factor in the formation and persistence of inflammation. In addition, shikonin also has anti-bacterial, anti-fungal, and anti-HIV effects. [0003] G...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K31/122A61P35/00
Inventor 凌昌全王园王丽娜张院辉
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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