BACKGROUND: Metabolomic analyses from our group and others have shown that tumors treated with glutamine antagonists (GA) exhibit robust accumulation of formylglycinamide ribonucleotide (FGAR), an intermediate in the de novo purine synthesis pathway. The increase in FGAR is attributed to the inhibition of the enzyme FGAR amidotransferase (FGAR-AT) that catalyzes the ATP-dependent amidation of FGAR to formylglycinamidine ribonucleotide (FGAM). While perturbation of this pathway resulting from GA therapy has long been recognized, no study has reported systematic quantitation and analyses of FGAR in plasma and tumors. OBJECTIVE: Herein, we aimed to evaluate the efficacy of our recently discovered tumor-targeted GA prodrug, GA-607 (isopropyl 2-(6-acetamido-2-(adamantane-1-carboxamido)hexanamido)-6-diazo-5-oxohexanoate), and demonstrate its target engagement by quantification of FGAR in plasma and tumors. METHODS: Efficacy and pharmacokinetics of GA-607 were evaluated in a murine EL4 lymphoma model followed by global tumor metabolomic analysis. Liquid chromatography-mass spectrometry (LC-MS) based methods employing the ion-pair chromatography approach were developed and utilized for quantitative FGAR analyses in plasma and tumors. RESULTS: GA-607 showed preferential tumor distribution and robust single-agent efficacy in a murine EL4 lymphoma model. While several metabolic pathways were perturbed by GA-607 treatment, FGAR showed the highest increase qualitatively. Using our newly developed sensitive and selective LC-MS method, we showed a robust >80- and >10- fold increase in tumor and plasma FGAR levels, respectively, with GA-607 treatment. CONCLUSION: These studies describe the importance of FGAR quantification following GA therapy in cancer and underscore its importance as a valuable pharmacodynamic marker in the preclinical and clinical development of GA therapies.

• Klíčová slova
• Glutamine antagonist, LC-MS., biomarker, cancer, formylglycinamide ribonucleotide, formylglycinamidine ribonucleotide, purine synthesis,
• MeSH
• biomarkery farmakologické analýza   metabolismus   MeSH
• chromatografie kapalinová metody   MeSH
• glutamin antagonisté a inhibitory   MeSH
• glycin analogy a deriváty   analýza   metabolismus   MeSH
• hmotnostní spektrometrie metody   MeSH
• metabolické sítě a dráhy účinky léků   MeSH
• myši MeSH
• nádorové biomarkery analýza   metabolismus   MeSH
• nádory * farmakoterapie   metabolismus   MeSH
• ribonukleotidy * analýza   metabolismus   MeSH
• vyvíjení léků metody   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biomarkery farmakologické MeSH
• glutamin MeSH
• glycin MeSH
• nádorové biomarkery MeSH
• phosphoribosyl-N-formylglycineamide MeSH Prohlížeč
• ribonukleotidy * MeSH

Aberrant excitatory neurotransmission associated with overproduction of glutamate has been implicated in the development of HIV-associated neurocognitive disorders (HAND). The glutamine antagonist 6-diazo-5-oxo-l-norleucine (DON, 14) attenuates glutamate synthesis in HIV-infected microglia/macrophages, offering therapeutic potential for HAND. We show that 14 prevents manifestation of spatial memory deficits in chimeric EcoHIV-infected mice, a model of HAND. 14 is not clinically available, however, because its development was hampered by peripheral toxicities. We describe the synthesis of several substituted N-(pivaloyloxy)alkoxy-carbonyl prodrugs of 14 designed to circulate inert in plasma and be taken up and biotransformed to 14 in the brain. The lead prodrug, isopropyl 6-diazo-5-oxo-2-(((phenyl(pivaloyloxy)methoxy)carbonyl)amino)hexanoate (13d), was stable in swine and human plasma but liberated 14 in swine brain homogenate. When dosed systemically in swine, 13d provided a 15-fold enhanced CSF-to-plasma ratio and a 9-fold enhanced brain-to-plasma ratio relative to 14, opening a possible clinical path for the treatment of HAND.

• MeSH
• aminokapronáty aplikace a dávkování   chemická syntéza   farmakologie   MeSH
• azosloučeniny aplikace a dávkování   chemická syntéza   farmakologie   MeSH
• diazooxonorleucin aplikace a dávkování   farmakologie   MeSH
• glutaminasa antagonisté a inhibitory   MeSH
• HIV infekce komplikace   MeSH
• krev metabolismus   MeSH
• kyselina glutamová metabolismus   MeSH
• lidé MeSH
• mozek metabolismus   MeSH
• myši inbrední C57BL MeSH
• neurokognitivní poruchy farmakoterapie   etiologie   MeSH
• nootropní látky aplikace a dávkování   chemická syntéza   farmakologie   MeSH
• prasata MeSH
• prekurzory léčiv aplikace a dávkování   chemická syntéza   farmakologie   MeSH
• stabilita léku MeSH
• virová nálož účinky léků   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aminokapronáty MeSH
• azosloučeniny MeSH
• diazooxonorleucin MeSH
• glutaminasa MeSH
• isopropyl 6-diazo-5-oxo-2-(((phenyl(pivaloyloxy)methoxy)carbonyl)amino)hexanoate MeSH Prohlížeč
• kyselina glutamová MeSH
• nootropní látky MeSH
• prekurzory léčiv MeSH