AC484 pharmacokinetics were evaluated following single oral doses to groups of male C57Bl/6N mice (Charles River Laboratories). Mice were permitted free access to food and water. AC484 was administered as a solution in ethanol: PEG-400: Phosal 50 PG (Lipoid, LLC) (10:30:60, w/w) to mice (10 ml kg–1 dose volume). Groups of mice (3 per group) received a single 3, 10, 30 or 100 mg kg–1 oral dose by gavage. EDTA-preserved blood samples were obtained at 0.25, 0.5, 1, 3, 6, 9, 12 and 24 h after the dose in each mouse. Plasma was analysed as described for the low-dose study above.
Peg 400
Manufactured by Lipoid
PEG-400 is a polyethylene glycol with an average molecular weight of 400 g/mol. It is a clear, viscous liquid at room temperature. PEG-400 is commonly used as a solvent, humectant, and plasticizer in various pharmaceutical and cosmetic applications.
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3 protocols using peg 400
1
Oral Pharmacokinetics of AC484 in Mice
2
PDX Tumor Models for Drug Efficacy
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Tumors were established from dissociated PDX and tumor harboring mice were randomized into treatment and control cohorts of 6 animals when the tumor average size reached about 100 mm3 (single Afuresertib experiment) or individually distributed into different treatment groups when tumors reached about 100 mm3 (combination experiment). For Afuresertib treatment, drug (100 or 20 mg/kg) and vehicle were administered by oral gavage 5 times a week. Afuresertib (Selleck, S7521) was dissolved in 20% PEG-400 (Lipoid), 1% DMSO and 79% H2O. For trametinib (ApexBio, A3018) treatment, drug (0.5 mg/kg) was dissolved in 4% DMSO/ corn oil and administered by i.p. injection five times a week. Tumor size was measured three times a week using a caliper and mouse weight was measured twice a week. No mice needed to be euthanized due to severe body weight loss (>20% than baseline).
3
Combination Therapy for PDX Tumors
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Tumors were established from dissociated PDX and tumor harboring mice were randomized into treatment and control cohorts of 6 animals when the tumor average size reached about 100 mm3. ABT-263 (ApexBio, A3007) was dissolved in a mixture of 10% EtOH, 30% PEG-400 (Lipoid) and 60% phosal 50 PG (Sigma Aldrich) and was given orally (100 mg/kg) alone or 1.5 h after standard chemotherapy in combination experiments. Compound solution was prepared fresh before drug administration and any remaining solution was routinely stored at +4°C for no longer than 1 wk. Vincristine (Teva) was injected intravenously (i.v.) (0.5 mg/kg). Tumor size was measured 3 times a wk using a caliper and mouse weight was measured twice a wk. No mice needed to be euthanized due to severe body weight loss (>20% than baseline). All animal experiments were performed under license of the authorities of the Kt. Zürich (206/15).
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