1 or 0.3 μg/kg) and its corresponding OVX-vehicle control group VE-821 data. Unless stated otherwise, all the procedures described
below were the same for both experiments. Our study used female cynomolgus monkeys, at least 9 years of age from China. Animals were housed in pairs, and were fed Certified Hi-Fiber Primate Diet 5 K91 (PMI Nutrition International Inc., St. Louis, MO, USA) or 2050C Teklad Certified Global 20% Protein Primate Diet (Harlan Laboratories, Indianapolis, IN, USA) twice daily, with food supplements of Prima-Treats (Bio Serv, Frenchtown, NJ, USA) and/or fresh fruit, and had free access to water. The animal room environment was controlled, with settings targeted at a temperature of 22 ± 3 °C, humidity of 50 ± 20%, 12 h light and 12 h dark photoperiod, and 12 air changes per hour. Animals were acclimated for a period of approximately 5 to 6 weeks after receipt prior to the baseline monitoring period. Only animals considered in good health, with established menses, minimal skeletal abnormalities, closed epiphyseal plates, and normal baseline serum/urine chemistry panels were included (data not presented). A total of 40 animals were used in the study. For each experiment, 20 animals were randomly assigned to either the OVX-vehicle control group or the eldecalcitol group. Following completion of baseline bone mineral density (BMD) measurements by dual-energy
X-ray absorptiometry (DXA), groups were balanced to ensure that age, body IPI-145 weight, whole body bone mineral content (BMC), and lumbar spine BMD were equivalent across groups within each experiment. The surgical procedures were performed under general anesthesia which included an injection of glycopyrrolate, ketamine hydrochloride, and xylazine, followed by isoflurane gas. Daily oral gavage treatment commenced the day following ovariectomy, with either OVX-vehicle control (OVX-Veh1 Thymidine kinase or OVX-Veh2) or eldecalcitol (at 0.1 μg/kg in the first experiment or 0.3 μg/kg in the second experiment), and continued for 6 months.
Body weights were monitored weekly and food consumption assessed twice daily (data not presented). Animals were fasted overnight prior to sample collection. Blood samples were collected prior to surgery and prior to termination after 6 months of treatment. For the second experiment, samples were also collected at month 3. After serum separation, serum calcium and phosphorus were analyzed with a Roche Hitachi 717 Chemistry Analyzer (Hitachi High-Tech Corp., Tokyo, Japan). The serum bone-specific alkaline phosphatase (BAP) was measured with a Metra BAP ELISA kit (Quidel Corporation, San Diego, CA, USA) or Tandem-R-Ostase IRMA assay kit (Hybritech Inc., San Diego, CA, USA), and the serum collagen C-telopeptide (CTX) was measured with a CrossLaps ELISA kit (Nordic Bioscience Diagnostics, Herlev, Denmark).