Alkylaminophenol and GPR17 Agonist for Glioblastoma Therapy: A Combinational Approach for Enhanced Cell Death Activity

Abstract

Drug resistance and tumor heterogeneity limits the therapeutic efficacy in treating glioblastoma, an aggressive infiltrative type of brain tumor. GBM cells develops resistance against chemotherapeutic agent, temozolomide (TMZ), which leads to the failure in treatment strategies. This enduring challenge of GBM drug resistance could be rational by combinatorial targeted therapy. Here, we evaluated the combinatorial effect of phenolic compound (2-(3,4-dihydroquinolin-1(2H)-yl)(p-tolyl)methyl)phenol (THTMP), GPR17 agonist 2-({5-[3-(Morpholine-4-sulfonyl)phenyl]-4-[4-(trifluoromethoxy)phenyl]-4H-1,2,4-triazol-3-yl}sulfanyl)-N-[4-(propan-2-yl)phenyl]acetamide (T0510.3657 or T0) with the frontline drug, TMZ, on the inhibition of GBM cells. Mesenchymal cell lines derived from patients’ tumors, MMK1 and JK2 were treated with the combination of THTMP + T0, THTMP + TMZ and T0 + TMZ. Cellular migration, invasion and clonogenicity assays were performed to check the migratory behavior and the ability to form colony of GBM cells. Mitochondrial membrane permeability (MMP) assay and intracellular calcium, [Ca2+]i, assay was done to comprehend the mechanism of apoptosis. Role of apoptosis-related signaling molecules was analyzed in the induction of programmed cell death. In vivo validation in the xenograft models further validates the preclinical efficacy of the combinatorial drug. GBM cells exert better synergistic effect when exposed to the cytotoxic concentration of THTMP + T0, than other combinations. It also inhibited tumor cell proliferation, migration, invasion, colony-forming ability and cell cycle progression in S phase, better than the other combinations. Moreover, the combination of THTMP + T0 profoundly increased the [Ca2+]i, reactive oxygen species in a time-dependent manner, thus affecting MMP and leading to apoptosis. The activation of intrinsic apoptotic pathway was regulated by the expression of Bcl-2, cleaved caspases-3, cytochrome c, HSP27, cIAP-1, cIAP-2, p53, and XIAP. The combinatorial drug showed promising anti-tumor efficacy in GBM xenograft model by reducing the tumor volume, suggesting it as an alternative drug to TMZ. Our findings indicate the coordinated administration of THTMP + T0 as an efficient therapy for inhibiting GBM cell proliferation.

Intrinsic and Microenvironmental Drivers of Glioblastoma Invasion

“…By coupling it with TMZ, it is also possible to further hamper GBM growth. In 2021, Doan et al [ 207 ] explored the use of a combinatorial drug regimen including TMZ, GPR-17 agonist 2-({5-[3-(Morpholine-4-sulfonyl)phenyl]-4-[4-(trifluoromethoxy)phenyl]-4H-1,2,4-triazol-3-yl}sulfanyl)-N-[4-(propan-2-yl)phenyl]acetamide, and alkylaminophenol against GBM. They saw that the combination of alkylaminophenol and GPR-17 agonist reduced GBM migration, invasion, and proliferation.…”

Anticancer Activity of Mannich Bases: A Review of Recent Literature

“…glioblastoma cells with a combination of Mannich base 67 (R=CH 3 ) (Figure 8), G protein‐coupled receptor 17 (GPR17) agonist T0510.3657 (AKos GmbH, Stuttgart, Germany) and frontline drug temozolomide has shown that the best synergistic effects were obtained with a combination of aminoalkylphenol 67 (R=CH 3 ) and T0510.3657, which inhibited tumor cell proliferation, cell migration, invasion, colony‐forming ability and cell cycle progression in S phase better than any other combination, and also showed promising anti‐tumor efficacy in glioblastoma xenograft model by reducing the tumor volume [84] . In addition, phenolic Mannich base 68 (Figure 8) can act as an agonist activating endogenous GPR17 in LN229 and SNB19 glioblastoma multiforme cells, and it had a cytotoxic effect on these cells in a dose‐dependent manner with IC 50 =85 μM for SNB19 cells [85] .…”