Extended Data Fig. 7: DAIA prodrugs are stable in plasma and liver microsomes, non-toxic to mammalian cells, do not disrupt mitochondrial membrane potential in C2C12 myoblasts and do not disrupt hERG function.
From: RETRACTED ARTICLE: IspH inhibitors kill Gram-negative bacteria and mobilize immune clearance
a, b, Nonlinear regression curves for degradation of prodrugs C23.28–TPP and C23.21–TPP and the appearance of the parent drugs C23.28 and C23.21 in the presence of human, pig and mouse plasma (a) or human, monkey and mouse liver microsomes (b). Drug and prodrug concentration measured by LC–MS and normalized on a standard curve. The half-lives (t 1/2) are calculated from respective curves. Data are mean ± s.e.m. of 3 independent experiments. c, d, Cytotoxicity of prodrug analogues on HepG2, RAW264.7 and Vero cells (c) and C2C12 myoblasts (d) measured at 24, 48 and 72 h by LDH release (n = 3 biological and 4 technical replicates). e, Effect of TPP and prodrugs C23.28–TPP and C23.47–TPP on mitochondrial membrane potential of C2C12 myoblasts, measured by tetramethyl rhodamine methyl ester (TMRM) fluorescence. CCCP, positive control (n = 3 biological and 4 technical replicates). f, Toxicity of C23.28–TPP, the carrier (6-hydroxyhexyl)triphenylphosphonium bromide and Me-TPP to hERG channel measured by automated Q patch assay; the normalized current response is plotted using nonlinear regression curves and the IC50 of respective compounds is calculated. Data are mean of 3 independent experiments ± s.e.m. Verapamil was used as the positive control and DMSO as the negative control.
