Biodistribution of oncolytic virus HSV-1 expressing human PD-1 antibody in cynomolgus macaques

doi:10.19803/j.1672-8629.20220181

Abstract

Abstract: Objective To establish a real-time quantitative PCR (qPCR) analysis method to investigate the biodistribution of oncolytic virus product HSV-1/hPD-1 in mice and cynomolgus macaques, and to detect samples collected during the concomitant biodistribution (shedding included) study in the process of investigating the repeat-dose toxicity in cynomolgus macaques. Methods According to the specific genetic sequences of drugs, primers and probes were designed and a proper PCR reaction system was established. Standard curves for detection were constructed and verified methodologically. Tissues and organs collected during repeat-dose toxicity experiments, including blood, gonads, kidneys, livers, lungs, hearts, brains, spleens, lymph nodes (mesenteric and inguinal), injection sites (skeletal muscle),tear gland, salivary gland, dorsal root ganglion, urine and feces, were subjected to qPCR detection. Results A universal standard curve named pMD18T-1093 was constructed, which could be used to quantitatively determine the copy number of the tested drugs in different tissues of mice and cynomolgus macaques. The tested drugs predominately remained at the injection sites and were sparsely distributed to the lung and brain before being cleared away by urine. Drug distribution was not detected in any other tissues of cynomolgus macaques. The amount of drugs located in target organs increased in a dose-dependent manner and decreased gradually with time rather than being accumulated in the body. Drugs detected in the brain might have been due to sample pollution. Conclusion A qPCR analysis method for preclinical biodistribution studies has been established in the study, which is expected to facilitate future studies on biodistribution of other drugs for gene therapy.

Key words: oncolytic virus, gene therapy, biodistribution, real-time quantitative PCR technique, repeat-dose toxicity, safety evaluation, cynomolgus macaques

CLC Number:

R969.1

WANG Xin, SUN Li, WANG Chao, LI Lulu, WANG Sanlong, LIU Jiajia, TIAN Chao, LI Xiaopeng, GENG Xingchao. Biodistribution of oncolytic virus HSV-1 expressing human PD-1 antibody in cynomolgus macaques[J]. Chinese Journal of Pharmacovigilance, 2023, 20(1): 12-18.

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