FIGURE 6.

The mTORC1/mTORC2 inhibitor and metformin combination facilitates tissue availability of the drugs: Tissue distribution of Torin 2 (A) and metformin (B) in an orthotopic mouse model injected with mTORC1/2 inhibitor or metformin. KPC cells were implanted into the pancreas in C57/BL6 mice. Once tumors became palpable, the mice were injected intraperitoneally daily for 12 d with Torin 2 (mTORC1 + mTORC2 inhibitor) at a high concentration (TH); Torin 2 at a low concentration (TL), metformin at a low concentration (ML), a combination of Torin 2 and metformin at low concentrations (TLML), or DMSO vehicle (control). At the end of the 12 d, the mice were sacrificed, and tissues were harvested and flash-frozen. Torin 2 and metformin concentrations were analyzed and quantified using LC-MS/MS. The data are reported as the average ± SD. One animal in the low-dose Torin 2 treatment group had a very high concentration of Torin 2 (∼4-fold higher) in all tissues and was considered an outlier and was not included in the statistical analysis. Drug concentrations were measured using LC-MS/MS. ANOVA test was performed using SPSS software (IBM) to determine the overall differences between groups. A post hoc test was employed to identify the pairs of groups that differed. Statistical significance was assigned as P < 0.05 if an overall statistical difference was detected between groups. Means with different letters were significantly different. ML, metformin at low dose; mTORC, mechanistic target of rapamycin complex; ns, nonsignificant results; TH, Torin 2 at high dose; TL, Torin 2 at low dose; TLML, Torin 2 and metformin at low doses.