Pharmacokinetics of Recombinant Anti-RANKL Humanized Monoclonal Antibody in Cynomolgus Monkey
WANG Meng-jia1, DONG Li-hou2, OU Lun3, WANG Bian-zhen3*, GE Zhi-qiang1*
1. School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China; 2. State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences(Beijing), Beijing Institute of Lifeomics, Beijing 102206, China; 3. United-Power Pharma Tech Co., Ltd., Beijing 102206, China
Abstract：OBJECTIVE To develop and validate the in-house enzyme-linked immunosorbent assay (ELISA) to determine the concentration of recombinant anti-RANKL humanized monoclonal antibody (analyte, T) in cynomolgus serum to evaluate the pharmacokinetic behavior of T in cynomolgus monkey. METHODS A total of 32 healthy cynomolgus monkeys were selected and randomly divided into 4 dosing groups, which were subcutaneously injected 0.5, 3 and 18 mg·kg-1 of T, and intravenously injected 3 mg·kg-1 of T, respectively. The concentrations of T in the serum samples was detected by the validated ELISA, and the measurements were imported into WinNonLin® v6.4 (Pharsight Inc.) to calculate the pharmacokinetic parameters. RESULTS The quantitative range of the in-hourse ELISA assay is 31.3 to 2 000 ng·mL-1, and the intra-and inter-batch precision and accuracy are in the range of 3.3% to 11.6% and -14.4% to 14.8%, respectively. The method was validated with acceptable accuracy, precision, specificity, sensitivity, dilution linearity, hook effect, selectivity, parallelism, and stability required by regulations. After subcutaneous administration of 0.5, 3 and 18 mg·kg-1 of T and intravenous injection of 3 mg·kg-1 T, concentrations in each group reach peak concentrations at 24-72, 10-72, 24-168 and 0.083 3-4 h, respectively; the peak concentrations (ρmax) are (7.57±0.872), (37.9±6.72), (250±35.5) μg·mL-1 and (87.2±13.8) μg·mL-1; areas under concentration-time curve (AUC0-t) are (1 630±496), (8 810±3 800), (94 700±34 900) and (12 000±4 370) μg·h·mL-1. The absolute bioavailability of the 3 mg·kg-1 subcutaneous injection group is 73.4%. CONCLUSION The in-house ELISA method can be used to support the pharmacokinetic study of T; the results of pharmacokinetic parameters indicate a linear pharmacokinetic behavior after subcutaneous injection of 0.5-18 mg·kg-1 of analyte in cynomolgus monkeys.
ABRAHAMSEN B, OSMOND C, COOPER C. Life expectancy in patients treated for osteoporosis: observational cohort study using national danish prescription data[J]. J Bone & Mineral Res, 2015,30(9):1539-1553.
[2]
Chinese Society of Osteoporosis and Bone Mineral Research. Guidelines for diagnosis and treatment of primary osteoporosis (2017)[J]. Chin J Pract Int Med, 2017, 10(5):413-443.
[3]
KONG X H, NIU Y B, LI Y H, et al. Progress of research on the relationship between OPG/RANK/RANKL system and osteoporosis [J]. Life Sci Res, 2011, 15(1):80-85.
[4]
STEVENS J A, RUDD R A. The impact of decreasing US hip fracture rates on future hip fracture estimates[J]. Osteoporosis Int, 2013, 24(10):2725-2728.
[5]
RYSER M D, QU Y, KOMAROVA S V, et al. Osteoprotegerin in bone metastases: mathematical solution to the puzzle[J]. PLoS Comput Biol, 2012, 8(10):e1002703.
[6]
LIU Z P. Osteoprotegerin influences the bone resorption activity of osteoclasts[J]. Int J Mol Med, 2013, 31(6):1411-1417.
[7]
HOFBAUER, LORENZ C. Clinical implications of the osteoprotegerin/RANKL/RANK system for bone and vascular diseases[J]. JAMA, 2004, 292(4):490-495.
[8]
XIONG Q, ZHANG L C, ZHANG L H, et al. Effects of recombinant human osteoprotegerin and recombinant RANK protein on the differentiation of osteoclast precursors[J]. China J Orthop Trauma(中国骨伤), 2013,26(4):324-327.
[9]
MIDORI, NAKAMURA, NOBUYUKI, et al. Osteoporosis and RANKL signal[J]. Clin Calcium, 2011, 21(8):1149-1155.
[10]
KIESEL L, KOHL A. Role of the RANK/RANKL pathway in breast cancer[J]. Maturitas, 2016, 86:10-16.
[11]
LIU W, XU C, ZHAO H, et al. Osteoprotegerin induces apoptosis of osteoclasts and osteoclast precursor cells via the Fas/Fas ligand pathway[J]. PLoS One, 2015, 10(11):e0142519.
[12]
LI X N. Researching progress of OPG-RANK-RANKL signaling pathway [J]. Henan Med Res, 2018(1):1802-1804.
[13]
LACEY D L, BOYLE W J, SIMONET W S, et al. Bench to bedside: elucidation of the OPG-RANK-RANKL pathway and the development of denosumab[J]. Nat Rev Drug Dis, 2012,11(5):401-419.
[14]
LI Z Y, LI Y K. Researching progress and application of OPG-RANK-RANKL signaling pathway in osteoporosis [J]. Chin J Geriatr Orthop Rehabil (Elect Ed), 2017, 3(2):124-128.
[15]
WANG R D, JIA P, TANG H. The role of receptor activator of nuclear factor-KB/receptor activator of nuclear factor-KB ligand/osteoprotegerin signaling pathway in the mechanism of breast cancer bone metastasis[J]. Chin J Exp Surgery, 2019, 36(5):982-986.
[16]
CHOW, CHEUNG L T. Giant cell rich osteosarcoma revisited-diagnostic criteria and histopathologic patterns, Ki67, CDK4, and MDM2 expression, changes in response to bisphosphonate and denosumab treatment[J]. Virchows Archiv, 2016, 468(6):741-755.
[17]
AMGEN INCORPORATED. Highlights of prescrbing information[EB/OL]. 2010. [2020-05-30]. http://pi.amgen.com/united_states/prolia/prolia_pi.pdf.
[18]
EUROPEAN MEDICI NES AGENCY. Guideline on bioanalytical method validation[J]. Drug Eval Res, 2012,35(5):396-398.
[19]
SHENG Y C, HE Y C, YANG J, et al. Evaluation on the methodology of dose proportionality study in clinical pharmacokinetics[J]. Chin J Clin Pharm, 2010, 26(5):376-381.
[20]
WANG C, CHEN L, YANG J. Application of confidence interval in the assessment of dose proportionality[J]. J China Pharm Univ(中药科大学学报), 2008, 39(6):534-537.
[21]
REGINSTER J Y. Antifracture efficacy of currently available therapies for postmenopausal osteoporosis[J]. Drugs, 2011, 71(1): 65-78.
[22]
ZHU R F, DING X L, MIAO L Y. Advances in therapeutic drug monitoring of monoclonal antibody drugs[J]. Chin Pharm J(中国药学杂志), 2018, 53(20):1714-1717.
[23]
JI S M, ZHU J. Application of target-mediated drug disposition (TMDD) models in the development of monoclonal antibody[J]. Chin J Clin Pharm Ther, 2018,23(3):241-246.
[24]
GIDAL B E, CLARK A M, ANDERS B, et al. The application of half-life in clinical decision making: comparison of the pharmacokinetics of extended-release topiramate (USL255) and immediate-release topiramate[J]. Epilepsy Res, 2016,129:26.
[25]
BOXENBAUM H, BATTLE M. Effective half-life in clinical pharmacology[J]. J Clin Pharmacol, 1995, 35(8):763-766.
[26]
SAHIN S, BENET L Z. The operational multiple dosing half-life: a key to defining drug accumulation in patients and to designing extended release dosage forms[J]. Pharm Res, 2008, 25(12):2869-2877.
[27]
ZHANG C F, XIE H T, PAN G Y. Absorption, distribution, metabolism, excretion and toxicity of biologics and its application in pharmacokinetic modeling [J]. Acta Pharm Sin(药学学报), 2016, 51(8):1202-1208.
[28]
MOULD D R, GREEN B. Pharmacokinetics and pharmaco-dynamics of monoclonal antibodies[J]. Bio Drugs, 2010, 24(1):23-39.
2021, Vol. 56 
