Manufacture Of Metal-organic Framework For Transcellular Protein Delivery And Macrophage Transformation For Tumor Immunotherapy

In recent years,nano-scale metal-organic framework materials material(MOF)have been widely used in the biological fields.We developed a pH-responsive MOF material capable of lysosomal escape function to carry natural SOD-1 that is not easily endocytosed by cells to achieve protein transmembrane transport.In addition,we modify the surface groups of the above-mentioned MOF materials to develop a MOF drug with a highly active proton-absorbable lysosomal inhibitor,which causes the successful transformation of M2 tumor-associated macrophages into M1 macrophage cells which suppresses tumor growth to achieve therapeutic effect.A stable nanometer colloid of MOF-PGMA(EA)/SOD-60 is formed and MOF-PGMA(EA)combines a pH-responsive and surface-rich positive charge to integrate with SOD-1.The XRD result shows that the MOF core is successfully formed.Acid incubation experiment proves that SOD-1 has successfully achieved charge reversal and maintained enzyme activity.The results of dynamic light scattering(DLS)and Zeta potential experiment prove that the particle size of the nanometer colloid is about 36.8nm and the potential is+19.8mV.Cell uptake and lysosomal escape experiments have proven that the nanometer collid has a higher cell uptake rate and lysosomal escape ability than the blank group and the control group.The extracellular SOD-1 enzyme activity test has showed that the nano-assembly has a higher ability to inhibit ROS.In summary,we have successfully prepared a nanometer colloid of MOF-PGMA(EA)/SOD-60 that can transmembrane.We then refine the above-mentioned MOF material to synthesize another MOF material that can successfully transform tumor-associated macrophages of M2 phenotype which promote tumor growth into M1 phenotype which suppresses tumor growth and also achieve capacity of lysosomal escape.Eventually,MOF-PGMA(DET)-CDM achieves immunotherapy of tumor cells.The successful synthesis of MOF-PGMA(DET)-CDM is verified by ~1H NMR and Fourier Transform Infrared Spectroscopy(FTIR).The results of dynamic light scattering(DLS)and Zeta potential experiments prove that the particle size of the nano particle is 45.0nm and the potential is-7.8mV.And the transmission electron microscope(TEM)and XRD experiments show that the crystal nuclei of MOF has been successfully formed and has good uniformity.Intracellular experiments confirmed that MOF-PGMA(DET)-CDM has good biocompatibility and capacity of charge reversal in the tumor microenvironment(pH=5.5)which is exposed a positive charge to promote cellular uptake and confirme lysosomal escape ability.Western Blotting,RT-qPCR,Flow Cytometry and ELISA experiments detect that marker cytokines of M2 phenotype and M1 phenotype have demonstrated and MOF-PGMA(DET)-CDM has stronger phenotypic regulation than chloroquine(A drug has been proven to have the ability of transform phenotypes of macrophages),we have successfully synthesized a more efficient proton-absorbent lysosomal inhibitor.