title: Antitumor efficacy of intermittent treatment schedules with the rapamycin derivative RAD001 correlates with prolonged inactivation of ribosomal protein S6 kinase 1 in peripheral blood mononuclear cells.
abstract: The orally bioavailable rapamycin derivative RAD001 (everolimus) targets the mammalian target of rapamycin pathway and possesses potent immunosuppressive and anticancer activities. Here, the antitumor activity of RAD001 was evaluated in the CA20948 syngeneic rat pancreatic tumor model. RAD001 demonstrated dose-dependent antitumor activity with daily and weekly administration schedules; statistically significant antitumor effects were observed with 2.5 and 0.5 mg/kg RAD001 administered daily [treated tumor versus control tumor size (T/C), 23% and 23-30%, respectively], with 3-5 mg/kg RAD001 administered once weekly (T/C, 14-36%), or with 5 mg/kg RAD001 administered twice weekly (T/C, 36%). These schedules were well tolerated and exhibited antitumor potency similar to that of the cytotoxic agent 5-fluorouracil (T/C, 23%). Moreover, the efficacy of intermittent treatment schedules suggests a therapeutic window allowing differentiation of antitumor activity from the immunosuppressive properties of this agent. Detailed biochemical profiling of mammalian target of rapamycin signaling in tumors, skin, and peripheral blood mononuclear cells (PBMCs), after a single administration of 5 mg/kg RAD001, indicated that RAD001 treatment blocked phosphorylation of the translational repressor eukaryotic initiation factor 4E-binding protein 1 and inactivated the translational activator ribosomal protein S6 kinase 1 (S6K1). The efficacy of intermittent treatment schedules was associated with prolonged inactivation of S6K1 in tumors and surrogate tissues (> or =72 h). Furthermore, detailed analysis of the dose dependency of weekly treatment schedules demonstrated a correlation between antitumor efficacy and prolonged effects (> or =7 days) on PBMC-derived S6K1 activity. Analysis of human PBMCs revealed that S6K1 also underwent a concentration-dependent inactivation after RAD001 treatment ex vivo (>95% inactivation with 20 nM RAD001). In contrast, human PBMC-derived eukaryotic initiation factor 4E-binding protein 1 was present predominantly in the hypophosphorylated form and was unaffected by RAD001 treatment. Taken together, these results demonstrate a correlation between the antitumor efficacy of intermittent RAD001 treatment schedules and prolonged S6K1 inactivation in PBMCs and suggest that long-term monitoring of PBMC-derived S6K1 activity levels could be used for assessing RAD001 treatment schedules in cancer patients.
authors.affiliation: Novartis Institutes for BioMedical Research Basel, Novartis Pharma AG, Basel, Switzerland.
authors.forename: Anne
authors.initials: A
authors.lastname: Boulay
authors.forename: Sabine
authors.initials: S
authors.lastname: Zumstein-Mecker
authors.forename: Christine
authors.initials: C
authors.lastname: Stephan
authors.forename: Iwan
authors.initials: I
authors.lastname: Beuvink
authors.forename: Frederic
authors.initials: F
authors.lastname: Zilbermann
authors.forename: Roland
authors.initials: R
authors.lastname: Haller
authors.forename: Sonja
authors.initials: S
authors.lastname: Tobler
authors.forename: Christoph
authors.initials: C
authors.lastname: Heusser
authors.forename: Terence
authors.initials: T
authors.lastname: O'Reilly
authors.forename: Barbara
authors.initials: B
authors.lastname: Stolz
authors.forename: Andreas
authors.initials: A
authors.lastname: Marti
authors.forename: George
authors.initials: G
authors.lastname: Thomas
authors.forename: Heidi A
authors.initials: HA
authors.lastname: Lane
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004334:Drug Administration Schedule; D016903:Drug Monitoring / Q000379:methods; D000068338:Everolimus; D006801:Humans; D007166:Immunosuppressive Agents / Q000008:administration & dosage / Q000627:therapeutic use*; D007963:Leukocytes, Mononuclear / Q000201:enzymology*; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008297:Male; D010190:Pancreatic Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000276:immunology; D051381:Rats; D011917:Rats, Inbred Lew; D038744:Ribosomal Protein S6 Kinases, 90-kDa / Q000037:antagonists & inhibitors* / Q000097:blood; D020123:Sirolimus / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000627:therapeutic use*; D013997:Time Factors; D014407:Tumor Cells, Cultured
chemical_list: D007166:Immunosuppressive Agents; D000068338:Everolimus; C069329:RPS6KA1 protein, human; D038744:Ribosomal Protein S6 Kinases, 90-kDa; C512963:Rps6ka1 protein, rat; D020123:Sirolimus