Application of nano-selenium serving as CIK (cytokine-induced killer) cell sensitizer

A technology of nano-selenium and sensitizer, which is applied in the field of medicine, can solve problems such as easy tolerance, and achieve the effect of low toxicity and inhibition of proliferation

Active Publication Date: 2017-05-24
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Liver cancer is prone to resistance t

Method used

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  • Application of nano-selenium serving as CIK (cytokine-induced killer) cell sensitizer
  • Application of nano-selenium serving as CIK (cytokine-induced killer) cell sensitizer
  • Application of nano-selenium serving as CIK (cytokine-induced killer) cell sensitizer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Preparation and characterization of embodiment 1 nano-selenium

[0076] The present embodiment has synthesized nano-selenium by oxidation-reduction method, utilizes vitamin c (Vc, Aladdin) to reduce selenous acid (H 2 SeO 3 ) or sodium selenite (Na 2 SeO 3 ), reuse PEG (COOH-PEG5000), polyvinylpyrrolidone (PVP, Aladdin, P110607), chitosan (CS, Sinopharm Chemical Reagent Co., Ltd.) or pachyphyllin (CMP, Hunan Butian Pharmaceutical Co., Ltd.) Modified nano-selenium, that is, prepared (PEG-SeNPs, PVP-SeNPs, CS-SeNPs, CMP-SeNPs), and detected their influence on the survival rate of CIK cells and the mechanism of co-action with CIK cells.

[0077] 1. Preparation of S-SeNPs:

[0078] Specifically, at 4°C, 1 mL of 10 mM H 2 SeO 3 solution or Na 2 SeO 3 Put the solution (final concentration 1-5mM) into a small beaker, slowly add 1mL 40mM Vc (the molar ratio of Vc to Se is 3-10:1), then dilute to 5mL with ultrapure water and react at 4°C for 0.5-24 Hours, use a dialysis...

Embodiment 2

[0093] Example 2 Study on anti-hepatoma tumor activity of S-SeNPs and CIK cells in vitro

[0094] 1. Culture of HepG2 cells and CIK cells

[0095] (1) HepG2 cells were purchased from the American Standard Cell Collection (ATCC), cultured with complete medium (DMEM, Gibco), and 10% (v / v) fetal bovine serum (FBS, Gibco) and 1% (v / v) double antibody (penicillin-streptomycin mixed solution, Gibco), cultured in a humidified incubator with 5% (v / v) carbon dioxide at 37°C.

[0096] (2) For the cultivation of CIK cells, 50 ml of peripheral blood from normal people was aseptically collected, anticoagulated with heparin, and sent back to the laboratory for subsequent operations in a sterile ultra-clean bench. The blood sample was diluted with PBS buffer (concentration: 0.01M, pH 7.4, the same below) equal to the volume of the blood sample. Add human lymphocyte separation medium (Shenzhen Dakowei Biological Engineering Co., Ltd.) equal to the volume of the undiluted blood sample into a...

Embodiment 3

[0103] Example 3 Inhibitory effect of S-SeNPs and CIK on the growth of human liver cancer cell HepG2 xenograft tumor in nude mice

[0104] 1. Prepare the model

[0105] Collect the human liver cancer cells HepG2 cultured in vitro (the culture method is the same as in Example 2), count, and adjust the concentration of the cell suspension to be 1 × 10 7 cells / ml, inoculate 0.15mL cell suspension subcutaneously in the right axillary of nude mice (BALB / c-nu mice, 28 days old, Beijing Huafukang Biotechnology Co., Ltd.);

[0106] 2. Grouping and administration

[0107] Use a vernier caliper to measure the diameter of the transplanted tumor in nude mice, and wait until the tumor grows to 75-100mm 3 Then the animals were randomly divided into 5 groups, 10 in each group. Start administration at the same time, see group and administration plan for administration plan, use the method of measuring tumor diameter, observe (once every 3 days) the anti-tumor effect of test substance, mice...

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Abstract

The invention discloses an application of nano-selenium serving as a CIK (cytokine-induced killer) cell sensitizer. The application of the nano-selenium serving as the CIK cell sensitizer refers to applying the nano-selenium as the CIK cell sensitizer to assist CIK cells. The nano-selenium provided by the invention is at least one of pure nano-selenium, polyethylene glycol modified nano-selenium, polyvinylpyrrolidone modified nano-selenium, chitosan modified nano-selenium, polysaccharide modified nano-selenium, folic acid modified tumor-targeting nano-selenium and transferrin modified tumor-targeting nano-selenium; and the CIK cells are obtained by performing mononuclear cell separation on human peripheral blood, bone marrow or cord blood, and performing induced culture by using a CD-3 antibody and IFN-gamma and IL-2 in vitro. The nano-selenium sensitizer disclosed by the invention has low-price and easily-available raw materials, synthesis and purification steps are strong in operability, and the synthesis scale can be appropriately enlarged by virtue of process optimization, thereby achieving commercialization and application of the medicine.

Description

technical field [0001] The invention belongs to the technical field of medicine, and particularly relates to the application of nano-selenium as a CIK cell sensitizer. Background technique [0002] In recent years, the good news of tumor immunotherapy has continued. At present, it has demonstrated strong anti-tumor activity in the treatment of some tumor types such as melanoma, gastric cancer and non-small cell lung cancer. FDA (Food and Drug Administration, FDA) approved clinical application. Due to its excellent efficacy and innovation, tumor immunotherapy was named the most important scientific breakthrough of the year by Science in 2013 [Science, 2013, 342:1666]. Tumor immunotherapy is expected to become an innovation in the field of tumor treatment following surgery, chemotherapy, radiotherapy, and targeted therapy [Yao Yang, et al. Nanoscale Research Letters, 2016, 11:285]. CIK cells refer to autologous cytokine-induced killer cells, which are powerful anti-tumor or ...

Claims

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Application Information

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IPC IPC(8): A61K33/04A61K35/17A61P35/00C01B19/02B82Y40/00B82Y30/00
CPCA61K33/04A61K35/17C01B19/02C01P2004/04C01P2004/52A61K2300/00
Inventor 陈填烽刘婷贺利贞
Owner JINAN UNIVERSITY
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