Limits...
P7170, a novel inhibitor of mTORC1/mTORC2 and Activin receptor-like Kinase 1 (ALK1) inhibits the growth of non small cell lung cancer.

Venkatesha VA, Joshi A, Venkataraman M, Sonawane V, Bhatia D, Tannu P, Bose J, Choudhari S, Srivastava A, Pandey PK, Lad VJ, Sangana R, Ahmed T, Damre A, Deore V, Sahu B, Kumar S, Sharma S, Agarwal VR - Mol. Cancer (2014)

Bottom Line: P7170 at a well-tolerated daily dose of 20 mg/kg significantly inhibited the growth of NSCLC xenografts independent of different mutations (EGFR, KRAS, or PIK3CA) or sensitivity to erlotinib.Pharmacokinetic-pharmacodynamic (PK-PD) analysis showed sub-micro molar tumor concentrations along with mTORC1/C2 inhibition.Our results provide evidence of antitumor activity of P7170 in the erlotinib -sensitive and -insensitive models of NSCLC.

View Article: PubMed Central - PubMed

Affiliation: Piramal Life Sciences Ltd, # 1 Nirlon Complex, Off: Western Express Highway, Goregaon (East), Mumbai, Maharashtra 400063, India. veena_828@yahoo.com.

ABSTRACT

Background: Lung cancer is the major cause of cancer-related deaths and many cases of Non Small Cell Lung Cancer (NSCLC), a common type of lung cancer, have frequent genetic/oncogenic activation of EGFR, KRAS, PIK3CA, BRAF, and others that drive tumor growth. Some patients though initially respond, but later develop resistance to erlotinib/gefitinib with no option except for cytotoxic therapy. Therefore, development of novel targeted therapeutics is imperative to provide improved survival benefit for NSCLC patients. The mTOR cell survival pathway is activated in naïve, or in response to targeted therapies in NSCLC.

Methods: We have discovered P7170, a small molecule inhibitor of mTORC1/mTORC2/ALK1 and investigated its antitumor efficacy using various in vitro and in vivo models of human NSCLC.

Results: P7170 inhibited the phosphorylation of AKT, S6 and 4EBP1 (substrates for mTORC2 and mTORC1) levels by 80-100% and growth of NSCLC cells. P7170 inhibited anchorage-independent colony formation of NSCLC patient tumor-derived cells subsistent of disease sub-types. The compound also induced apoptosis in NSCLC cell lines. P7170 at a well-tolerated daily dose of 20 mg/kg significantly inhibited the growth of NSCLC xenografts independent of different mutations (EGFR, KRAS, or PIK3CA) or sensitivity to erlotinib. Pharmacokinetic-pharmacodynamic (PK-PD) analysis showed sub-micro molar tumor concentrations along with mTORC1/C2 inhibition.

Conclusions: Our results provide evidence of antitumor activity of P7170 in the erlotinib -sensitive and -insensitive models of NSCLC.

Show MeSH

Related in: MedlinePlus

Anti-tumor efficacy of P7170 correlated with tumor pharmacokinetic and pharmacodynamic parameters. (A) Tumor volume (mm3) of H460 xenografts in SCID mice was measured in P7170 treatment groups twice weekly. (B) For pharmacodynamic studies, P7170 was administered into tumor bearing mice once daily consecutively for 3 days. Plasma and tumors samples were collected at 1, 4, 8, 24 h post-P7170-last dose (minimum 3 mice per time point) for estimating P7170 concentrations by LC-MS/MS. P7170 concentrations for each dose (1, 5, 10, or 20 mg/kg) were calculated based on the AUC (area under curve) constructed using concentrations at 1, 4, 8, 24 h post-P7170-last dose. Statistical significance: *p < 0.05 (C) Representative protein western blots showing response to the treatment in the experiment (B). (D) In the same experiment as in (B), part of tumor tissues was processed for western blotting analyses of proteins using specific antibodies; protein band intensities were normalized to respective loading controls. The band intensities of pS6 were plotted against the absolute plasma or tumor P7170 concentrations. The correlations of pS6 (S235/236) levels to plasma or tumor P7170 concentrations were performed using the non-compartmental analysis tool of WinNonlin Professional version 6.1: Predicted curve represents the trend in protein level derived from each tumors sample (closed circles) with respect to tumor concentrations of P7170.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4289333&req=5

Fig3: Anti-tumor efficacy of P7170 correlated with tumor pharmacokinetic and pharmacodynamic parameters. (A) Tumor volume (mm3) of H460 xenografts in SCID mice was measured in P7170 treatment groups twice weekly. (B) For pharmacodynamic studies, P7170 was administered into tumor bearing mice once daily consecutively for 3 days. Plasma and tumors samples were collected at 1, 4, 8, 24 h post-P7170-last dose (minimum 3 mice per time point) for estimating P7170 concentrations by LC-MS/MS. P7170 concentrations for each dose (1, 5, 10, or 20 mg/kg) were calculated based on the AUC (area under curve) constructed using concentrations at 1, 4, 8, 24 h post-P7170-last dose. Statistical significance: *p < 0.05 (C) Representative protein western blots showing response to the treatment in the experiment (B). (D) In the same experiment as in (B), part of tumor tissues was processed for western blotting analyses of proteins using specific antibodies; protein band intensities were normalized to respective loading controls. The band intensities of pS6 were plotted against the absolute plasma or tumor P7170 concentrations. The correlations of pS6 (S235/236) levels to plasma or tumor P7170 concentrations were performed using the non-compartmental analysis tool of WinNonlin Professional version 6.1: Predicted curve represents the trend in protein level derived from each tumors sample (closed circles) with respect to tumor concentrations of P7170.

Mentions: In order to understand the antitumor efficacy of P7170 in erlotinib-sensitive and insensitive in vivo models, subcutaneous xenografts of NSCLC, using H1975, H1650, or H460 cell lines were established. Although H1650 and H1975 cell lines are equally responsive to erlotinib in vitro, literature [24, 25] suggests that H1975 xenografts are erlotinib-resistant. Treatment with P7170 resulted in a significant growth inhibition (70% TGI, p < 0.001, Table 2) of xenograft derived from H1650 erlotinib-sensitive cells [EGFR del E748-A750 (activating mutation); KRAS mutant; PIK3CA wild type] at 5 mg/kg, a dose much lower than its MTD. However, the same dose of P7170 was not very effective in mice bearing erlotinib-insensitive H1975 [EGFR T790M, L858R; KRAS wild type; PIK3CA wild type] xenografts, but treatment with a15 mg/kg dose resulted in a significant tumor growth inhibition (92% TGI, p < 0.001, Table 2). Interestingly, P7170 was efficacious (88% TGI at 20 mg/kg; 66% TGI at 10 mg/kg with p < 0.001, Table 2 and Figure 3A) in another model of erlotinib -insensitive H460 (EGFR wild type; KRAS mutant; PIK3CA mutant) xenografted animals. P7170 administration was well tolerated at doses up to 20 mg in different xenograft models (Additional file 4: Figure S4). These results provide evidence for the efficacy of P7170 in erlotinib-sensitive, and –insensitive NSCLC irrespective of known genetic mutations in these cells.Table 2


P7170, a novel inhibitor of mTORC1/mTORC2 and Activin receptor-like Kinase 1 (ALK1) inhibits the growth of non small cell lung cancer.

Venkatesha VA, Joshi A, Venkataraman M, Sonawane V, Bhatia D, Tannu P, Bose J, Choudhari S, Srivastava A, Pandey PK, Lad VJ, Sangana R, Ahmed T, Damre A, Deore V, Sahu B, Kumar S, Sharma S, Agarwal VR - Mol. Cancer (2014)

Anti-tumor efficacy of P7170 correlated with tumor pharmacokinetic and pharmacodynamic parameters. (A) Tumor volume (mm3) of H460 xenografts in SCID mice was measured in P7170 treatment groups twice weekly. (B) For pharmacodynamic studies, P7170 was administered into tumor bearing mice once daily consecutively for 3 days. Plasma and tumors samples were collected at 1, 4, 8, 24 h post-P7170-last dose (minimum 3 mice per time point) for estimating P7170 concentrations by LC-MS/MS. P7170 concentrations for each dose (1, 5, 10, or 20 mg/kg) were calculated based on the AUC (area under curve) constructed using concentrations at 1, 4, 8, 24 h post-P7170-last dose. Statistical significance: *p < 0.05 (C) Representative protein western blots showing response to the treatment in the experiment (B). (D) In the same experiment as in (B), part of tumor tissues was processed for western blotting analyses of proteins using specific antibodies; protein band intensities were normalized to respective loading controls. The band intensities of pS6 were plotted against the absolute plasma or tumor P7170 concentrations. The correlations of pS6 (S235/236) levels to plasma or tumor P7170 concentrations were performed using the non-compartmental analysis tool of WinNonlin Professional version 6.1: Predicted curve represents the trend in protein level derived from each tumors sample (closed circles) with respect to tumor concentrations of P7170.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4289333&req=5

Fig3: Anti-tumor efficacy of P7170 correlated with tumor pharmacokinetic and pharmacodynamic parameters. (A) Tumor volume (mm3) of H460 xenografts in SCID mice was measured in P7170 treatment groups twice weekly. (B) For pharmacodynamic studies, P7170 was administered into tumor bearing mice once daily consecutively for 3 days. Plasma and tumors samples were collected at 1, 4, 8, 24 h post-P7170-last dose (minimum 3 mice per time point) for estimating P7170 concentrations by LC-MS/MS. P7170 concentrations for each dose (1, 5, 10, or 20 mg/kg) were calculated based on the AUC (area under curve) constructed using concentrations at 1, 4, 8, 24 h post-P7170-last dose. Statistical significance: *p < 0.05 (C) Representative protein western blots showing response to the treatment in the experiment (B). (D) In the same experiment as in (B), part of tumor tissues was processed for western blotting analyses of proteins using specific antibodies; protein band intensities were normalized to respective loading controls. The band intensities of pS6 were plotted against the absolute plasma or tumor P7170 concentrations. The correlations of pS6 (S235/236) levels to plasma or tumor P7170 concentrations were performed using the non-compartmental analysis tool of WinNonlin Professional version 6.1: Predicted curve represents the trend in protein level derived from each tumors sample (closed circles) with respect to tumor concentrations of P7170.
Mentions: In order to understand the antitumor efficacy of P7170 in erlotinib-sensitive and insensitive in vivo models, subcutaneous xenografts of NSCLC, using H1975, H1650, or H460 cell lines were established. Although H1650 and H1975 cell lines are equally responsive to erlotinib in vitro, literature [24, 25] suggests that H1975 xenografts are erlotinib-resistant. Treatment with P7170 resulted in a significant growth inhibition (70% TGI, p < 0.001, Table 2) of xenograft derived from H1650 erlotinib-sensitive cells [EGFR del E748-A750 (activating mutation); KRAS mutant; PIK3CA wild type] at 5 mg/kg, a dose much lower than its MTD. However, the same dose of P7170 was not very effective in mice bearing erlotinib-insensitive H1975 [EGFR T790M, L858R; KRAS wild type; PIK3CA wild type] xenografts, but treatment with a15 mg/kg dose resulted in a significant tumor growth inhibition (92% TGI, p < 0.001, Table 2). Interestingly, P7170 was efficacious (88% TGI at 20 mg/kg; 66% TGI at 10 mg/kg with p < 0.001, Table 2 and Figure 3A) in another model of erlotinib -insensitive H460 (EGFR wild type; KRAS mutant; PIK3CA mutant) xenografted animals. P7170 administration was well tolerated at doses up to 20 mg in different xenograft models (Additional file 4: Figure S4). These results provide evidence for the efficacy of P7170 in erlotinib-sensitive, and –insensitive NSCLC irrespective of known genetic mutations in these cells.Table 2

Bottom Line: P7170 at a well-tolerated daily dose of 20 mg/kg significantly inhibited the growth of NSCLC xenografts independent of different mutations (EGFR, KRAS, or PIK3CA) or sensitivity to erlotinib.Pharmacokinetic-pharmacodynamic (PK-PD) analysis showed sub-micro molar tumor concentrations along with mTORC1/C2 inhibition.Our results provide evidence of antitumor activity of P7170 in the erlotinib -sensitive and -insensitive models of NSCLC.

View Article: PubMed Central - PubMed

Affiliation: Piramal Life Sciences Ltd, # 1 Nirlon Complex, Off: Western Express Highway, Goregaon (East), Mumbai, Maharashtra 400063, India. veena_828@yahoo.com.

ABSTRACT

Background: Lung cancer is the major cause of cancer-related deaths and many cases of Non Small Cell Lung Cancer (NSCLC), a common type of lung cancer, have frequent genetic/oncogenic activation of EGFR, KRAS, PIK3CA, BRAF, and others that drive tumor growth. Some patients though initially respond, but later develop resistance to erlotinib/gefitinib with no option except for cytotoxic therapy. Therefore, development of novel targeted therapeutics is imperative to provide improved survival benefit for NSCLC patients. The mTOR cell survival pathway is activated in naïve, or in response to targeted therapies in NSCLC.

Methods: We have discovered P7170, a small molecule inhibitor of mTORC1/mTORC2/ALK1 and investigated its antitumor efficacy using various in vitro and in vivo models of human NSCLC.

Results: P7170 inhibited the phosphorylation of AKT, S6 and 4EBP1 (substrates for mTORC2 and mTORC1) levels by 80-100% and growth of NSCLC cells. P7170 inhibited anchorage-independent colony formation of NSCLC patient tumor-derived cells subsistent of disease sub-types. The compound also induced apoptosis in NSCLC cell lines. P7170 at a well-tolerated daily dose of 20 mg/kg significantly inhibited the growth of NSCLC xenografts independent of different mutations (EGFR, KRAS, or PIK3CA) or sensitivity to erlotinib. Pharmacokinetic-pharmacodynamic (PK-PD) analysis showed sub-micro molar tumor concentrations along with mTORC1/C2 inhibition.

Conclusions: Our results provide evidence of antitumor activity of P7170 in the erlotinib -sensitive and -insensitive models of NSCLC.

Show MeSH
Related in: MedlinePlus