Anti-tumor pharmaceutical composition based on blocking PD-1/PD-L1 and application of anti-tumor pharmaceutical composition
An anti-tumor drug, PD-L1 technology, applied in the field of biomedicine, can solve the problem of reducing the reactivity and ambiguity of TRIB3 expression, and achieve the effect of increasing recruitment and killing activity and enhancing sensitization and reactivity
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
experiment example 1
[0037] Experimental example 1: Co-immunoprecipitation method proves that C646 can reduce the acetylation and protein expression of TRIB3 in tumor cells
[0038] Co-immunoprecipitation reagents are as follows:
[0039] Lysis solution A: 0.6057g Tris base, 1.7532g NaCl, 0.1017g MgCl2 6H2O, 0.0742g EDTA, 10mL glycerin, 10mL 10% NP40, add deionized water to 150mL, adjust the pH value to 7.6 with HCl, and adjust the volume to 191mL , mixed thoroughly, filtered through a 0.45 μm membrane filter, and stored at 4°C.
[0040] Lysis Solution B: 200 μL 2M β-glycerol phosphate, 4 mL 2.5M NaF, 2 mL 8 mM NaVO3, 2 mL 100 mMPMSF, 200 μL 1M DTT, 200 μL each of 1 mg / mL Leu, Pep and Apr, total volume 9 mL. The mother liquor was stored at -20°C. Before use, thaw the mother liquor of each component in liquid B, add it to liquid A according to the above composition ratio and mix well. Protein A / GPlus-Agarose was purchased from Santa Cruz, USA. The specific operation steps are as follows:
[00...
experiment example 2
[0044] Experimental example 2: The effect of the C646 combined drug composition based on immune checkpoint inhibition and reducing the expression of TRIB3 on tumor growth was studied by a mouse colon cancer MC38 tumor formation experiment.
[0045] First, Trib3 shRNA virus was used to construct MC38 stably knockdown TRIB3 cell line (MC38T3 KD ), and then use the Trib3K197R virus to infect the cell construction (MC38T3 KD / K197R OE ). K197R mutates the 197th lysine of TRIB3 to arginine, which can inhibit the ubiquitination of TRIB3 and maintain the stable and high expression of TRIB3.
[0046] The above mouse experiments were divided into 5 groups: isotype control antibody and control solvent treatment group (MC38), C646 alone treatment group (MC38), immune checkpoint PD-L1 blocking antibody alone treatment group (MC38), C646 / PD-L1 blocking Antibody combined treatment group (MC38) and C646 / PD-L1 blocking antibody combined treatment group (MC38T3 K D / K197R OE ).
[0047]...
experiment example 3
[0056] Experimental example 3: The effect of the C646 combined drug composition based on immune checkpoint inhibition and reducing the expression of TRIB3 on tumor growth was studied by the mouse liver cancer Hepa 1-6 tumor formation experiment.
[0057] Firstly, Trib3 shRNA virus was used to construct Hepa 1-6 stably knocking down TRIB3 cell line (Hepa1-6T3 KD ), and then use the Trib3K197R virus to infect the cell construction (Hepa 1-6T3 KD / K197R OE ). K197R mutates the 197th lysine of TRIB3 to arginine, which can inhibit the ubiquitination of TRIB3 and maintain the stable and high expression of TRIB3.
[0058] The above mouse experiments were divided into 5 groups: isotype control antibody and control solvent treatment group (Hepa 1-6), C646 treatment group alone (Hepa 1-6), immune checkpoint PD-L1 blocking antibody treatment group alone (Hepa 1-6). 6), C646 / PD-L1 blocking antibody combined treatment group (Hepa 1-6) and C646 / PD-L1 blocking antibody combined treatment ...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More