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Metabolism of GTI-2040, a phosphorothioate oligonucleotide antisense, using ion-pair reversed phase high performance liquid chromatography (HPLC) coupled with electrospray ion-trap mass spectrometry

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Abstract

GTI-2040 is a 20-mer phosphorothioate oligonucleotide, which is complementary to the messenger ribonucleic acid (mRNA) of the R2 subunit of ribonucleotide reductase. This study characterized both the in vivo and in vitro metabolism of GTI-2040. A highly specific ion-pair reversed-phase electrospray ionization (IP-RP-ESI) liquid chromatographymass spectrometry (LC-MS) method was used for the identification of GTI-2040 and metabolites from a variety of biological samples including exonuclease enzyme solutions, plasma, urine, mouse liver/kidney homogenates, and human liver microsomes. Progressively chain-shortened metabolites trucated from the 3′ terminal of GTI-2040 were detected in all of the evaluated biological samples. GTI-2040 was found to be a good substrate for 3′ but not 5′ exonuclease. While the pattern of n-1 chain-shortened 3′-exonucleolytic degradation was similar in the mouse liver and kidney homogenates, the latter was found to contain a larger number of shortenmers, the kidneys appeared to possess higher enzymatic reactivity toward GTI-2040. Thus, metabolism of GTI-2040 was found to occur in a variety of biological samples, mainly mediated by the 3′ exonuclease.

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Author information

Authors and Affiliations

  1. Division of Pharmaceutics, College of Pharmacy, The Ohio State University, 43210, Columbus, OH

    Xiaohui Wei, Zhongfa Liu, Hao Cheng, Zhiliang Xie & Kenneth K. Chan

  2. Pharmaceutical Research Institute, Bristol-Myers-Squibb Co, 08540, Princeton, NJ

    Guowei Dai

  3. The Comprehensive Cancer Center, The Ohio State University, 43210, Columbus, OH

    Guido Marcucci & Kenneth K. Chan

  4. Division of Hematology-Oncology, College of Medicine, The Ohio State University, 43210, Columbus, OH

    Guido Marcucci

  5. Room 308 OSU CCC, The Ohio State University, 410 West 12th Avenue, 43210, Columbus, OH

    Guido Marcucci

Authors
  1. Xiaohui Wei
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  2. Guowei Dai
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  3. Zhongfa Liu
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  4. Hao Cheng
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  5. Zhiliang Xie
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  6. Guido Marcucci
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  7. Kenneth K. Chan
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Correspondence to Kenneth K. Chan.

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Published: December 15, 2006

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Wei, X., Dai, G., Liu, Z. et al. Metabolism of GTI-2040, a phosphorothioate oligonucleotide antisense, using ion-pair reversed phase high performance liquid chromatography (HPLC) coupled with electrospray ion-trap mass spectrometry. AAPS J 8, 84 (2006). https://doi.org/10.1208/aapsj080484

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  • DOI: https://doi.org/10.1208/aapsj080484

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