PMX464

PMX464 (AW 464) is a thiol-reactive quinol compound and an inhibitor of the thioredoxin-thioredoxin reductase (Trx/TrxR) system, with an IC₅₀ value of 6.5 μM against TrxR. PMX464 acts by irreversibly inhibiting the thioredoxin (Trx-1) system, blocking HIF-1α transcriptional activation and the NF-κB inflammatory pathway, inducing apoptosis, attenuating platelet function and inhibiting thrombosis, and specifically disrupting the trypanothione antioxidant metabolic network in parasites. PMX464 is used in research concerning malignant tumors (colorectal cancer, breast cancer, renal cancer, and leukemia), pulmonary inflammation, African sleeping sickness, and antithrombotic applications^{[1][2][3][4][5][6][7]}. In Vitro:PMX464 (AW 464) (0.01-250 μM; 16-48 h) inhibits cell viability in MDA-MB-468 (IC₅₀ = 8.2 μM under hypoxia; IC₅₀ = 6.5 μM under normoxia), MDA-MB-231, and MDA-MB-435 cells, as well as RCC4 and 786-0 renal cancer cell lines, under both hypoxic and normoxic conditions; it also inhibits hypoxia-induced vascular endothelial growth factor (VEGF) secretion (IC₅₀ = 2.3 μM)^[1]. PMX464 (AW 464) (1-10 μM; 16 h) upregulates HIF-1α and HIF-2α protein expression in MDA-MB-468 and RCC4 cells (under hypoxic conditions) but downregulates the expression of HIF targets (CA-IX, BNIP3)^[1]. PMX464 (AW464) (30 min) inhibits the activity of the cell-free Trx1/TrxR1 system (IC₅₀ = 23 μM)^[2].
PMX464 (AW 464) (1 μM-100 μM; 6-8 h) induces apoptosis in HL-60 leukemia cells without triggering an oxidative stress burst^[2].
PMX464 (0.001-10 μM; 72 h) inhibits proliferation and colony formation in HT29 colorectal cancer cells (showing greater potency under hypoxic conditions) and induces G1/S cell cycle arrest and mild apoptosis^[3].
PMX464 (0.001-1 μM; 72 h) functionally inhibits Trx1 activity and feedback-upregulates TrxR1 protein expression in HT29 colorectal cancer cells, without altering Trx1 protein levels^[3].
PMX464 (0.5-1 μM; 72 h) inhibits proliferation in proliferating human umbilical vein endothelial cells (HUVECs) but shows weaker effects in quiescent HUVECs and MRCV fibroblasts^[3].
PMX464 (10-30 μM; 30 min pretreatment followed by 24 h stimulation) inhibits the expression of ICAM-1, CXCL8, and GM-CSF, as well as neutrophil adhesion and migration, in IL-1β- or TNF-α-stimulated A549 lung epithelial cells^[4].
PMX464 (30 μM; 30 min pretreatment) inhibits the NF-κB inflammatory pathway in IL-1β-stimulated A549 cells^[4].
PMX464 (0.6-1 μM; 30 min pretreatment followed by 24 h stimulation) inhibits cytokine-induced ICAM-1 expression in human lung microvascular endothelial cells (HLMVECs)^[4].
PMX464 (1-12 h) binds effectively to trypanosome-related targets T(SH)₂ (K_D = 0.43 μM), GSH (K_D = 6.2 μM), and L-cysteine (K_D = 27 μM)^[5].
PMX464 (73 h) effectively kills bloodstream-form Trypanosoma brucei (S427) (EC₅₀ = 77 nM) but has a minimal effect on normal human fibroblasts (MRC-5) (EC₅₀ = 175 nM)^[5].
PMX464 (30-90 min) inhibits the expression of trypanosome-related targets T(SH)₂ (IC₅₀ = 5.8 μM (30 min), IC₅₀ = 1.8 μM (90 min)), TDPX (IC₅₀ = 15.6 μM (30 min), IC₅₀ = 6.0 μM (90 min)), and TryP (IC₅₀ = 14.1 μM)^[5]. PMX464 (50-500 nM; 4-16 h) rapidly kills Trypanosoma brucei cells and depletes intracellular levels of trypanothione (T(SH)₂) and tryparedoxin peroxidase (TryP), without altering the protein levels of TryR, TryX, and TDPX^[5]. PMX464 (48-72 h) potently inhibits cell growth across the NCI-60 human cancer cell line panel, with mean GI₅₀ and LC₅₀ values of 0.23 μM and 3.39 μM, respectively; notably, the GI₅₀ value for HCT 116 cells is 0.11 μM^[6].
PMX464 (0.5-5 μM; 16 h) modulates target activity in a dose-dependent manner in HCT 116 cells^[6].
PMX464 (1-30 μM; 60 min) inhibits TrxR activity in a concentration-dependent manner in an in vitro rat recombinant TrxR enzyme system^[6].
PMX464 (15 μM; 5-60 min) inhibits TrxR activity in a time-dependent manner in an in vitro rat recombinant TrxR enzyme system^[6].
PMX464 (1-50 μM; 30 min) causes NADPH-dependent, irreversible inhibition of TrxR activity in an in vitro rat recombinant TrxR enzyme system; this inhibitory effect is partially attenuated by physiological concentrations of glutathione (GSH) in systems containing 0.5 mM or 1 mM GSH (HY-D0187). PMX464 demonstrates selectivity for mammalian TrxR^[6].
PMX464 (100 μM; 3-60 min) binds to and reduces free selenol groups in an in vitro rat recombinant TrxR enzyme system^[6].
PMX464 (30 µM; 30 min) reduces the extent of cell-surface free thiol labeling in washed human platelets^[7].
PMX464 (0.1-100 µM; 3 min) inhibits CRP-XL-induced and U46619 (HY-108566)-induced (0.1 µM) intracellular Ca2⁺ release in Fura-2-loaded (HY-101897) washed human platelets in a concentration-dependent manner, with IC₅₀ values of 10 µM and 20 µM, respectively^[7].
PMX464 (30 µM; 10-30 min) does not alter GPVI receptor surface levels in human whole blood^[7].
PMX464 (3-100 µM; 30 s-30 min) selectively inhibits CRP-XL-induced platelet aggregation in human platelet-rich plasma (PRP) in a time- and concentration-dependent manner; in human whole blood (under fluid shear conditions), it inhibits type I collagen surface thrombus formation and reduces surface coverage without causing GPIb degradation or surface shedding^[7].
PMX464 (30 µM) attenuates Ristocetin (HY-138053)-induced platelet agglutination in human platelet-rich plasma (PRP)^[7].
PMX464 attenuates CRP-XL-induced reactive oxygen species (ROS) generation in washed human platelets and inhibits clot retraction in human platelet-rich plasma (PRP)^[7].