Decitabine nano carrier and application thereof in preparing tumor fluorescence imaging agent

A nanocarrier, decitabine technology, applied in the field of materials and biology, can solve problems such as increasing the detoxification effect of active molecules and limiting the effect of iron apoptosis

Active Publication Date: 2020-06-09
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(GY. Liou, P. Storz, Reactive oxygen species cancer. Free Radic Res, 2010, 44(5), 479-496.) However, the abnormally high expression of glutathione (GSH) in RCC increases the reactive molecular detoxification, limiting the efficacy of ferroptosis

Method used

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  • Decitabine nano carrier and application thereof in preparing tumor fluorescence imaging agent
  • Decitabine nano carrier and application thereof in preparing tumor fluorescence imaging agent
  • Decitabine nano carrier and application thereof in preparing tumor fluorescence imaging agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] 1.1 Preparation and characterization of multifunctional decitabine nanocarriers (CNDHNNPs)

[0045] (1) Based on bovine hemoglobin, dissolve it in water to prepare a 10g / L hemoglobin solution, named solution A. (2) Take an appropriate amount of polylactic acid-glycolic acid copolymer PLGA and dissolve it in ethyl acetate to prepare a 18 mg / mL solution, which is named solution B. (3) Take an appropriate amount of Nile Red, prepare a 1g / L solution, and name it as solution C. (4) Take 400uL of solution A and solution C, put them in a 10ml centrifuge tube, add 2ml of solution B dropwise with refrigerated ultrasound at 4°C, mix well with a vortex oscillator, and ultrasonicate in an ice bath for 3mins with an interval of 20s in an ultrasonic breaker. The organic solvent was removed by a decompression freezing rotary evaporator, centrifuged at 10,000 rpm for 10 mins in a centrifuge, discarded the supernatant, washed 3 times with distilled water, freeze-dried using a lyophiliz...

Embodiment 2

[0046] Example 2 Multifunctional decitabine nanocarriers (CNDHNNPs) reverse hypoxia-mediated multidrug resistance

[0047] 2.1 Reduce the expression of hypoxia-inducible factor HIF-1α and its mediated drug resistance gene MDR1

[0048] (1) The hypoxic model of renal cell carcinoma 769-P was co-incubated with multifunctional decitabine nanocarriers (CNDHNNPs) for 48 hours, and the total RNA extraction kit (Tiangen Biochemical Technology (Beijing) Co., Ltd. ) according to the instructions to extract total cellular RNA from frozen human liver tissue, use a nucleic acid protein analyzer to measure the concentration and purity of RNA, and use a reverse transcription kit (Bao Bioengineering (Dalian) Co., Ltd.) to reverse transcribe the total RNA into cDNA.

[0049] (2) Design two rtPCR primers according to the DNA sequences of human HIF-1α and MDR1.

[0050] HIF-1α primer

[0051] 1) Forward: 5'-TCACCACAGGACAGTACAGGATGC-3'24

[0052] 2) Reverse: 5'-CCAGCAAAGTTAAAGCATCAGGTTCC-3'26 ...

Embodiment 3

[0076] Example 3 Multifunctional decitabine nanocarriers (CNDHNNPs) increase the expression of drug uptake transporter OCT2, and sensitize the effect of oxaliplatin against multidrug-resistant renal cell carcinoma

[0077] 3.1 Increased expression of cationic drug transporter OCT2

[0078] (1) The hypoxic model of renal cell carcinoma 769-P was co-incubated with multifunctional decitabine nanocarriers (CNDHNNPs) for 48 hours, and the total RNA extraction kit (Tiangen Biochemical Technology (Beijing) Co., Ltd. ) according to the instructions to extract total cellular RNA from frozen human liver tissue, use a nucleic acid protein analyzer to measure the concentration and purity of RNA, and use a reverse transcription kit (Bao Bioengineering (Dalian) Co., Ltd.) to reverse transcribe the total RNA into cDNA.

[0079] (2) Design two RT-PCR primers according to the DNA sequence of human SSL22A2.

[0080] OCT2 primer

[0081] 7) Forward: 5'-AAGAATGGGGATCACAATGG-3'20

[0082] 8) Re...

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Abstract

The invention provides a decitabine nano carrier and an application thereof in preparing a tumor fluorescence imaging agent. The decitabine nano carrier is prepared by a multiple emulsion method. Bovine hemoglobin, fluorescent dye Nile red, a polylactic acid-glycolic acid copolymer and the like are adopted for emulsification, then wrapping is performed by using chitosan loaded with decitabine andchitosan modified by nitrobenzooxadiazole (NBD), and sterilizing is performed to prepare freeze-dried powder, wherein the freeze-dried powder is dissolved in a water phase for oxygen loading before being used. The decitabine nano carrier is based on 769-P and OS-RC-2 kidney cancer cell lines, and a hypoxia cell model and a tumor-bearing mouse model are constructed. The results show that CNDHNNPs can improve the hypoxia microenvironment and reduce drug efflux, and decitabine can increase uptake of chemotherapeutic drugs and increase sensitization of chemotherapy. Peroxidation of lipid is increased by catalysis, and ferroptosis therapy is combined. The invention provides a novel scheme of hypoxia and hypermethylation combined ferroptosis of targeting tumor for renal cancer treatment, provides a drug delivery system, and realizes tumor fluorescence imaging. The decitabine nano carrier is reasonable in design and high in repeatability.

Description

technical field [0001] The invention belongs to the field of materials and biotechnology, and relates to a decitabine nanocarrier and its application in the preparation of tumor fluorescence imaging agents. The drug oxaliplatin is used in the treatment of hypoxia-mediated drug-resistant renal cancer, and has applications in tumor fluorescence imaging effects and other aspects. [0002] technical background [0003] Multi-drug resistance of kidney cancer is a major factor hindering the treatment of kidney cancer, (R.L.Siegel, K.D.Miller, A.Jemal, Cancer statistics, 2018.CA Cancer J Clin.2018,68(1),7-30.) Abnormal Epigenetic alterations and the hypoxic tumor microenvironment are considered important factors of chemotherapy resistance. (C.Holohan, S.VanSchaeybroeck, D.B. Longley, P.G.Johnston, Cancer drug resistance: an evolving paradigm. Nat Rev Cancer. 2013, 13, 714-726.) One of the epigenetic mechanisms of renal cancer chemotherapy resistance, namely SLC22A2 (coding OCT2 pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00A61K31/706A61K31/555A61P35/00
CPCA61K31/555A61K31/706A61K49/0028A61K49/0056A61K49/0093A61P35/00A61K2300/00
Inventor 曾苏王泽阳余露山
Owner ZHEJIANG UNIV
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