A composition capable of sensitizing NK cells and its application

A technology of NK cells and drugs, which can be used in drug combinations, medical preparations with non-active ingredients, and medical preparations containing active ingredients, etc. It can solve the problems of cytotoxicity of immune cells, inhibit the potential of immunotherapy, etc., and achieve immunity. Inhibitory effect, promote immune killing effect, enhance expression effect

Active Publication Date: 2022-04-29
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition to the efficacy of chemoimmunotherapy, most of these drugs were found to be cytotoxic to infused immune cells, which inhibited the potential of immunotherapy

Method used

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  • A composition capable of sensitizing NK cells and its application
  • A composition capable of sensitizing NK cells and its application
  • A composition capable of sensitizing NK cells and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Preparation of nanoemulsion loaded with selenocystine and TGF-β inhibitor

[0056] 1. Preparation

[0057] Weigh 2.0g of Poloxamer 188, 2.4g of olive oil, 2.4g of absolute ethanol, and 10mg of SeC into a beaker, then add 100μL of the prepared SB505124 stock solution (10mM), and stir at room temperature at 600rpm until the system becomes homogeneous phase, then drop by drop into 10mL of deionized water, and continue to stir for 30min to prepare colostrum. Subsequently, the prepared colostrum was transferred to a high-pressure homogenizer for homogenization step by step: first at a homogenization pressure of 250 Bar, homogenization for 3 minutes, and a sampling frequency of 40 Hz; then high-pressure homogenization, that is, the homogenization pressure was Homogenize for 20 minutes under the condition of 1200Bar, and the sampling frequency is 40Hz, and collect samples after homogenization. No SeC and SB505124 were added during the preparation process, and the o...

Embodiment 2

[0065] Embodiment 2 screening model drug test

[0066] NK cells and T cells extracted from tumor patients: 50 mL of peripheral blood was collected from the peripheral blood of tumor patient volunteers in a tertiary hospital in Guangdong, and peripheral blood mononuclear cells (PBMC) were extracted by Ficoll density gradient method; the cells were suspended in fresh ALYS505NK -In AC medium, adjust the density to 2×10 6 cells / mL, cultivated in a cell culture box for one day; then add 1000 U / mL recombinant human interleukin-2 (rhIL-2) and 10% autologous plasma to the cell culture medium, and continue to incubate for one day; the third day Add 30 mL of fresh ALYS505NK-EX medium, add 1000 U / mL rhIL-2 and rhIL-5, and incubate for two days; add 60 mL of fresh ALYS505NK-EX culture medium on the fifth day, add 1000 U / mL rhIL-2, and continue to incubate for 2 days; On the 7th to 14th day of culture, according to the color change of the culture medium, add double fresh serum-free medium...

Embodiment 3

[0076] Example 3 The Study of SSB NMs Stimulating NK92

[0077] (1) The combined effect of SSB NMs and NK92 and the ratio of NK92 cells added are set as follows:

[0078] Group 1: After pre-incubating different proportions of NK92 cells with different concentrations of SSB NMs for 12 hours, the two were added to MDA-MB-231 cells at the same time; the drug concentrations were 1 μM, 4 μM, 8 μM, 16 μM and 32 μM, and The addition amount was 100 μl / well. Tumor cells were counted according to the number when they were plated, and the amount when plated was 4×10 3 pcs / well (96-well plate). Effector cell:tumor cell (E:T) ratios were 2.5:1, 5:1, 10:1 and 20:1, respectively.

[0079] Group 2: MDA-MB-231 cells were pretreated with different concentrations of SSB NMs for 12 hours, and then NK92 cells were added in different proportions; the concentration of drugs and the ratio of effector cells: tumor cells were set the same as in Group 1.

[0080] Group 3: Different concentrations of...

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Abstract

The invention discloses a composition capable of sensitizing NK cells and its application. The composition includes selenocystine and a TGF-beta inhibitor. The selenocystine and the TGF-β inhibitor have the effect of synergistically sensitizing NK cells, and the composition can be used to prepare NK cell-sensitizing drugs. On the one hand, the nanoemulsion prepared by this composition effectively increases the solubility and stability of SeC; on the other hand, the nano-sized system can enhance the expression of NKG2D ligands on the surface of tumor cells, and relieve the immunosuppressive effect in the tumor microenvironment , effectively promoting the immune killing effect of NK cells on metastatic MDA‑MB‑231 cells.

Description

technical field [0001] The invention relates to a composition, in particular to a composition capable of sensitizing NK cells and its application. Background technique [0002] In recent years, cancer immunotherapy has provided an important breakthrough in the treatment of cancer. Strategies that stimulate the body's innate immune system to eliminate tumors and that enhance the immune response through immunomodulators (such as anti-PD-1 / PDL-1 antibodies and CTLA-4 therapy) have been shown to improve treatment outcomes for many types of cancer. However, most of these approaches focus on the delivery of engineered tumor vaccines or exogenous antigens due to the immunosuppression of the tumor microenvironment, such as inactivation of antigen-presenting cells, hyperactivation of regulatory T cells, immunosuppressive cytokine-induced cd 8+ T / NK cell dysfunction and loss of HLA expression in tumor cells, these approaches are highly variable and limited. In addition, immune chec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/107A61K47/44A61K47/10A61K45/06A61K31/198A61K31/4439A61P35/00A61P37/04
CPCA61K9/1075A61K47/44A61K47/10A61K45/06A61K31/198A61K31/4439A61P35/00A61P37/04A61K2300/00
Inventor 陈填烽刘畅李晓玲
Owner JINAN UNIVERSITY
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