Thiopurine Pathway

The thiopurine drugs are purine antimetabolites widely used in the treatment of acute lymphoblastic leukemia, autoimmune disorders (e.g., Crohn's disease and Rheumatoid Arthritis) and of organ transplant recipients. As inactive prodrugs, 6-mercaptopurine (6-MP), 6-thioguanine (6-TG) and azathioprine (AZA) require intracellular activation, catalyzed by multiple enzymes, to exert cytotoxicity. The first step in AZA activation is the release of 6-MP , which involves glutathione transferases and non-enzymatic conversion. Transport of 6-MP into the cell involves SLC28A2, SLC28A3, SLC29A1 and SLC29A2. After uptake, 6-MP is converted into thioinosine monophosphate (TIMP) by hypoxanthine guanine phosphoribosyl transferase (HPRT1), with 5-phospho-D-ribose-1-pyrophosphate (PRPP) as the phosphoribosyl donor. In a similar way, 6-TG can be converted into thioguanosine monophosphate (TGMP). TIMP can be converted into TGMP in two steps: first, thioxanthosine monophosphate (TXMP) is formed by inositol monophosphate dehydrogenase (IMPDH); second, TGMP is formed by guanosine monophosphate synthetase (GMPS). Subsequently, TGMP can be converted into thioguanine nucleotide diphosphates (TGDP) and triphosphates (TGTP). Cytotoxic effects of thiopurine drugs are achieved through incorporation of thio-deoxyguanosine triphosphate (TdGTP) into DNA and of thioguanosine triphosphate (TGTP) into RNA, by inhibition of de novo purine synthesis by methylmercaptopurine nucleotides (MeMPR) and by inhibition of Rac1 (this inhibition induces apoptosis in activated T cells). The TdGTP incorporation inhibits the function of several enzymes involved in DNA replication and repair, and induces DNA damage such as single strand-breaks, DNA-protein cross-links and chromatid exchanges. The pathway that leads to synthesis of active metabolites is in competition with inactivation pathways catalyzed by xanthine oxidase (XDH) or the polymorphic thiopurine methyltransferase (TPMT).

Abbreviations:
6-meMPR: 6-methylmercaptopurine riboside , 8-OHTG: 8-hydroxythioguanine , ABCC4: Multidrug resistance-associated protein 4 , ABCC5: Multidrug resistance-associated protein 5 , ADA: Adenosine deaminase , ADK: Adenosine kinase , AdoHcy: S-adenosylhomocysteine , AdoMet: S-adenosyl-methionine , AHCY: S-adenosylhomocysteine hydrolase , AOX1: Aldehyde oxidase 1 , GAR: Glycinamide ribotide , GART: Phosphoribosylglycinamide formyltransferase , GMPS: Guanosine monophosphate synthetase , GSTA1: glutathione-S-transferase A1 , GSTA2: glutathione-S-transferase A2 , GSTM1: glutathione-S-transferase M1 , HPRT1: Hypoxanthine guanine phosphoribosyltransferase 1 , IMPDH1: Inosine monophosphate dehydrogenase type 1 , ITPA: inosine triphosphatase pyrophosphatase , meMP: methylmercaptopurine , meTGMP: Methyl-thioguanosine monophosphate , meTIMP: Methyl-thioinosine monophosphate , MPR: 6-mercaptopurine riboside , NT5E: Nucleotidase , ecto-5-prime , PPAT: Phosphoribosyl pyrophosphate amidotransferase , PRA: 5-phosphoribosylamine , PRPP: 5-phospho-D-ribose- 1-pyrophosphate , PRPS1: Phosphoribosyl pyrophosphate synthetase 1 , RAC1: ras-related C3 botulinum toxin substrate 1 , SLC28A2: solute carrier family 28 , member 2 , SLC28A3: solute carrier family 28 , member 3 , SLC29A1: solute carrier family 29 , member 1 , SLC29A2: solute carrier family 29 , member 2 , TdGDP: 6-thio-deoxy-guanosine diphosphate , TdGTP: 6-thio-deoxy-guanosine triphosphate , TGDP: 6-thio-guanosine diphosphate , TGMP: Thioguanosine monophosphate , TGTP: 6-thio-guanosine triphosphate , TIMP: Thioinosine monophosphate , TITP: thioinosine triphosphate , TPMT: Thiopurine S-methyltransferase , TXMP: Thioxanthosine monophosphate , XDH: Xanthine dehydrogenase