Extended Data Fig. 4 ∣. Portimine A mouse pharmacokinetic properties and fast acting in vitro cell-based target engagement properties based on compound wash-out.

(a) Pharmacokinetic studies of mouse intraperitoneal (i.p.) and oral (p.o.) administration for portimine. Data presented as mean ± s.d. (n = 3 biologically independent samples). (b-d) Washout experiment performed in b) Jurkat c) MC38 and d) HT-1080 cells showed exposure-dependent decrease in IC₅₀, reveals PA has a fast-acting cytotoxicity mechanism. Data presented as mean ± s.d. (n = 3 biologically independent samples). (e) FACS analysis of apoptosis using annexin V/ eFluor 780 viability dye after PA and analogs (24 h) treatment in MC38 cell line. Data presented as mean ± s.d. of biological replicated experiments (n = 3 biologically independent samples). Statistical analysis was performed using one-way ANOVA analysis with multiple comparisons test analysis; P-values are shown. (f) Kaplan-Myer survival curve of WT C57BL/6 MC38 tumor-bearing mice (n = 6 mice per group) after treatment with PA 0.3 or 1 mg kg⁻¹ intraperitoneally.