Antitumor activity of nab-sirolimus versus mTOR inhibitors temsirolimus, sirolimus, and everolimus in A549 NSCLC xenografts.

person Shihe Hou Aadi Bioscience, Pacific Palisades, CA info_outline Shihe Hou, Maria Zalath, Andrew Kwon, Edward Spindler

Authors person Shihe Hou Aadi Bioscience, Pacific Palisades, CA info_outline Shihe Hou, Maria Zalath, Andrew Kwon, Edward Spindler Organizations Aadi Bioscience, Pacific Palisades, CA Abstract Disclosures Research Funding No funding sources reported Background: The mTORC1 pathway, often activated by mutations in genes like TSC1, TSC2, PIK3CA, PTEN, STK11, and KEAP1 , plays a pivotal role in cancer progression; in some settings, alterations in STK11 and KEAP1 may be associated with treatment resistance and poor prognosis. Despite the broad importance of the mTORC1 pathway in cancer cell growth and survival, mTOR inhibitors (mTORis) temsirolimus (TEM), sirolimus (SIRO), and everolimus (EVE) have limited clinical application in the cancer setting. nab -Sirolimus, an injectable form of albumin-bound SIRO that leverages unique transport properties of albumin known to increase tumor drug accumulation, is approved for treatment of malignant PEComa. Here, we report the antitumor activity of nab- sirolimus in comparison to other mTORis in A549 NSCLC ( KRAS G12S, STK11 Q37*, and KEAP1 G333C) xenografts, and the correlation of antitumor activity, tumor PK profile, and mTOR pathway suppression. Methods: Athymic mice bearing subcutaneous A549 NSCLC xenografts were treated with either saline or equal weekly doses of nab- sirolimus (administered intravenously [IV]; 5 or 15 mg/kg/week), TEM (IV; 5 mg/kg/week), or SIRO or EVE (both oral; 15 mg/kg/week). Tumors were harvested at different time points after mTORi treatment and analyzed for tumor drug levels (LC-MS/MS) and mTOR pathway biomarkers (pS6 and p4EBP1) via western blot (WB). Results: In A549 xenografts, nab- sirolimus (IV) treatment resulted in significantly greater suppression of tumor growth compared with TEM (IV), SIRO (oral), and EVE (oral) (Table). Average intratumoral drug concentrations 24 hours after IV mTORi treatment were significantly higher with nab- sirolimus (420-539 ng/g) compared with TEM (TEM [parent] 34.9 ng/g; SIRO [active metabolite measured from TEM] 13.2 ng/g) and compared with 7-day steady-state concentrations for oral SIRO (17 ng/g) and EVE (10 ng/g). WB confirmed that nab- sirolimus consistently inhibited mTOR targets pS6 and p4EBP1, whereas TEM, SIRO, and EVE were less effective. Conclusions: nab -Sirolimus resulted in significantly higher intratumoral drug concentration, stronger inhibition of mTOR targets, and greater antitumor activity compared to other IV and oral mTORis in a KRAS/STK11/KEAP1 mutated NSCLC xenograft model. These results support further clinical evaluation of nab -sirolimus as a single agent or in combination with other therapeutic agents in oncology. Dosing and antitumor activity of nab- Sirolimus vs TEM, SIRO, and EVE. Treatment N Weekly dose (mg/kg/wk) Schedule, Route Day vs Saline TGI vs saline (%) Tumor Growth vs nab- Sirolimus (ANOVA), P -value nab- Sirolimus 8 5 2.5 mg/kg, 2x/wk, IV 35 78.3 - TEM 8 5 2.5 mg/kg, 2x/wk, IV 35 45.0 0.01 nab- Sirolimus 8 15 7.5 mg/kg, 2x/wk, IV 35 94.2 - SIRO 8 15 2.14 mg/kg, daily, oral 35 30.0 0.0007 nab- Sirolimus 10 15 7.5 mg/kg, 2x/wk, IV 29 95.4 - EVE 10 15 2.14 mg/kg, daily, oral 29 68.3 0.07

Pre-clinical