LAUSR

ABSTRACT Drug‐induced kidney injury (DIKI) is a major concern in drug risk assessment given its clinical importance and the absence of a sensitive/specific method of diagnosis. Pharmaceutical regulatory agencies have qualified and issued letters of support for new biomarkers to better evaluate DIKI in nonclinical toxicity and clinical studies. Additional efforts have focused on drug‐induced phospholipidosis (DIPL) and its potential link with collateral renal damage. The combined use of urinary biomarkers is an efficient way to evaluate renal safety in nonclinical and clinical studies. Eight FDA/EMA/PMDA qualified (or supported) urinary biomarkers, including kidney injury molecule‐1 (KIM‐1), &bgr;2‐microglobulin (B2M), clusterin (CLU), cystatin C (CysC), trefoil factor 3 (TFF3), neutrophil gelatinase‐associated lipocalin (NGAL), osteopontin (OPN), and alpha‐glutathione S‐transferase (&agr;‐GST), were quantified by multiplex UPLC‐MS/MS in a repeat dose study of gentamicin in rats. Rats administered gentamicin at 100 mg/kg/day for 2 weeks developed renal lesions detected by histopathology. Biomarkers of tubular damage (CLU, KIM‐1, OPN) increased 9.8, 34.7, and 35.6‐fold (relative to concurrent controls), respectively, after 2 weeks of dosing. Biomarkers of glomerular damage and/or impairment of tubular reabsorption (CysC, B2M) increased 11.7 and 22.6‐fold. NGAL and &agr;‐GST increased <3‐fold after 2 weeks of dosing. TFF3 was comparable to concurrent controls. The elevated biomarker concentrations met PSTC threshold criteria and were consistent with mechanisms of gentamicin nephrotoxicity. Increased urinary di‐22:6‐BMP indicated concomitant DIPL as confirmed by TEM. This work provides evidence supporting the combined use of the DIKI biomarker panel and di‐22:6‐BMP as a biomarker of DIPL in drug risk assessment. Graphical abstract Figure. No caption available. HighlightsEvaluate drug‐induced kidney injury and phospholipidosis with safety biomarkersAssess kidney injury with eight FDA/EMA/PMDA qualified/ supported urine proteinsMonitor drug‐induced phospholipidosis with urine di‐22:6‐BMPEffectively monitor the locale/sequence of injury with a combined biomarker approachEvaluate nonclinical and clinical drug candidates with potential safety risks