Abstract
Mucopolysaccharidosis type II, commonly called Hunter syndrome, is a rare X-linked recessive disease caused by the deficiency of the lysosomal enzyme iduronate-2-sulphatase (I2S). A deficiency of I2S causes an abnormal glycosaminoglycans accumulation in the body’s cells. Although enzyme replacement therapy is the standard therapy, adeno-associated viruses (AAV)–based gene therapy could provide a single-dose solution to achieve a prolonged and constant enzyme level to improve patient’s quality of life. Currently, there is no integrated regulatory guidance to describe the bioanalytical assay strategy to support gene therapy products. Herein, we describe the streamlined strategy to validate/qualify the transgene protein and its enzymatic activity assays. The method validation for the I2S quantification in serum and method qualification in tissues was performed to support the mouse GLP toxicological study. Standard curves for I2S quantification ranged from 2.00 to 50.0 μg/mL in serum and 6.25 to 400 ng/mL in the surrogate matrix. Acceptable precision, accuracy, and parallelism in the tissues were demonstrated. To assess the function of the transgene protein, fit-for-purpose method qualification for the I2S enzyme activity in serum was performed. The observed data indicated that the enzymatic activity in serum increased dose-dependently in the lower I2S concentration range. The highest I2S transgene protein was observed in the liver among tissue measured, and its expression level was maintained up to 91 days after the administration of rAAV8 with a codon-optimized human I2S. In conclusion, the multifaceted bioanalytical method for I2S and its enzymatic activity were established to assess gene therapy products in Hunter syndrome.
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Acknowledgements
The authors would like to thank Dr. Vivian Choi at Global Gene Therapy and Dr. Nancy Chen at Global Drug Metabolism and Pharmacokinetics, Takeda Development Center Americas Inc., for their helpful discussion and constructive comments on this manuscript. The authors would like to thank Mr. Robert Crooker, Ms. Kathleen Palmieri at Rare Disease Drug Discovery Unit, and Dr. Bhanu Dasari at Global Gene Therapy for contributing to the method development. The authors would like to thank Dr. Kevin Cawthern, Ms. Amy Turner, Mr. Scott Goldberg, Dr. Ivan Ivanov, Ms. Lourdes Guerra, Mr. Mark Saewert, Dr. Bhanumathy Cunnigaiper, and Mr. Kenneth Tang at Pharmaceutical Development Services, Smithers, for their help with generating study reports or participating in method qualification/validation and sample analysis.
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This study was funded by Takeda Development Center Americas Inc. No writing assistance was utilized in the production of this manuscript.
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Franchi, P.M., Kulagina, N., Ilinskaya, A. et al. Multifaceted Approach for Quantification and Enzymatic Activity of Iduronate-2-Sulfatase to Support Developing Gene Therapy for Hunter Syndrome. AAPS J 25, 61 (2023). https://doi.org/10.1208/s12248-023-00821-6
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DOI: https://doi.org/10.1208/s12248-023-00821-6