Extended Data Fig. 10: In vivo profiling of TH9028 and TH9619 in mouse models.
(a) Kaplan-Meier curve showing overall survival of NOG mice with HL-60 IV xenograft tumors after treatment with TH9028 (30 mg/kg bid) or TH9619 (30 mg/kg bid) versus vehicle control, on low folate diet, n = 6–7 per group. P = 0.001 calculated using a one-way Mantel-Cox log-rank test. (b) Body weight measurements. Mouse body weight was monitored for two weeks before treatment start and followed throughout the study. At day 37 of treatment the remaining animals regained weight after diet shift from low folate diet to standard chow diet. Data are displayed as means + SEM, n = 7 (TH9028) or 8 (vehicle/TH9619) per group. Dotted line indicates treatment initiation on day 1. (c) Plasma concentration levels at 15 min and 30 min after the last administration of TH9028 (30 mg/kg bid) or TH9619 (30 mg/kg bid) in xenograft mice, n = 3 per group. Data are displayed as means ± SD. (d) Intratumor levels of TH9619, monoglutaminated TH9619 (TH9619-1G), 5-meTHF and thymidine at the time of sacrifice, measured by LC-MS and displayed as means ± SD, n = 2 (vehicle) or 3 (treated). * P < 0.05 calculated by unpaired two-tailed t-test (PTH9619 = 0.0449, tTH9619 = 3.324, DFTH9619 = 1; PTH9619-1G = 0.0168, tTH9619-1G = 4.841, DFTH9619-1G = 1). (e) Target engagement of MTHFD2 in tumor samples analyzed using CETSA. Graphs show the non-denatured target fraction from TH9028- and TH9619-treated animals and vehicle controls, displayed as means ± SD, n = 2 (vehicle) or 3 (treated). MTHFD2 ΔTm (TH9028) = 6.4 °C, MTHFD2 ΔTm (TH9619) = 5.9 °C.
