William DEMAIO - Academia.edu (original) (raw)
Papers by William DEMAIO
Bazedoxifene (BZA) acetate, a novel estrogen receptor modulator being developed for the preventio... more Bazedoxifene (BZA) acetate, a novel estrogen receptor modulator being developed for the prevention and treatment of post-menopausal osteoporosis, undergoes extensive metabolism in women following oral administration. In this study, the in vitro metabolism of [ 14 C]BZA was determined in human hepatocytes and hepatic and intestinal microsomes, and the UGT isozymes involved in the glucuronidation of BZA were identified. In addition, BZA was evaluated for its potential as a substrate of Pglycoprotein (P-gp) transporter in Caco-2 cell monolayers. BZA was metabolized to two mono-glucuronides, BZA-4'-glucuronide and BZA-5-glucuronide in hepatocytes and in liver and intestinal microsomes including jejunum, duodenum and ileum. Both BZA-4'glucuronide and BZA-5-glucuronide were major metabolites in the intestinal microsomes, while BZA-4'-glucuronide was the predominant metabolite in liver microsomes and hepatocytes. The kinetic parameters of BZA-4'-glucuronide formation were determined in liver, duodenum, and jejunum microsomes and with UGT 1A1, 1A8, 1A10, the most active UGT isoforms involved in the glucuronidation of BZA, while those of BZA-5-glucuronide were determined with all the enzyme systems except in liver microsomes and in UGT 1A1 as the formation of the BZA-5-glucuronide was too low. K m values in liver, duodenum, and jejunum microsomes and UGT 1A1, 1A8, 1A10, were similar and ranged from 5.1 to 33.1 µM for BZA-4'-glucuronide formation and 2.5 to 11.1 µM for BZA-5-glucuronide formation. V max values ranged from 0.8 to 2.9 nmol/min/mg protein for BZA-4'-glucuronide and from 0.1 to 1.2 nmol/min/mg protein for BZA-5-glucuronide. In Caco-2 cells, BZA appeared to be a P-gp substrate.
dm d.aspetjournals.org D ow nloaded from DMD #31179
PloS one, 2016
Parkinson's disease (PD) is a neurodegenerative aging disorder in which postmortem PD brain e... more Parkinson's disease (PD) is a neurodegenerative aging disorder in which postmortem PD brain exhibits neuroinflammation, as well as synucleinopathy-associated protein phosphatase 2A (PP2A) enzymatic activity loss. Based on our translational research, we began evaluating the PD-repurposing-potential of an anti-inflammatory, neuroprotective, and PP2A stimulatory oral drug that is FDA-approved for multiple sclerosis, FTY720 (fingolimod, Gilenya®). We also designed two new FTY720 analogues, FTY720-C2 and FTY720-Mitoxy, with modifications that affect drug potency and mitochondrial localization, respectively. Herein, we describe the metabolic stability and metabolic profiling of FTY720-C2 and FTY720-Mitoxy in liver microsomes and hepatocytes. Using mouse, rat, dog, monkey, and human liver microsomes the intrinsic clearance of FTY720-C2 was 22.5, 79.5, 6.0, 20.2 and 18.3 μL/min/mg; and for FTY720-Mitoxy was 1.8, 7.8, 1.4, 135.0 and 17.5 μL/min/mg, respectively. In hepatocytes, both FTY7...
Desvenlafaxine succinate (DVS) is an orally active serotonin and norepinephrine reuptake inhibito... more Desvenlafaxine succinate (DVS) is an orally active serotonin and norepinephrine reuptake inhibitor in development for treatment of major depressive disorder and vasomotor symptoms. Desvenlafaxine (DV) is a racemic mixture of the major active metabolite of venlafaxine. The metabolite profiles of 14C-DVS were investigated in plasma (1, 4 and 8 hr post-dose) and excreta (up to 24 hr) collected from laboratory animals (mice, rats and dogs) following a single oral administration. In plasma samples collected at 1 and 4 hr post-dose, parent compound represented ≤12% of the radioactivity in mice and rats, and ≤22% of the radioactivity in dogs. For all species, parent compound was not detected by radiochromatography in plasma samples collected at 8 hr post-dose. The major metabolite in plasma from each species was the O-glucuronide conjugate of DV, which represented greater than 76% of the circulating radioactivity. Additional minor oxidative and N-desmethyl metabolites represented less than...
SAX-187 (2-{1-[(6-chloroimidazo[2,1-b][1,3]thiazol-5-yl)sulfonyl]-1H-indol-3-yl}ethanamine hydroc... more SAX-187 (2-{1-[(6-chloroimidazo[2,1-b][1,3]thiazol-5-yl)sulfonyl]-1H-indol-3-yl}ethanamine hydrochloride) is a novel chemical compound with selective agonist activity at the 5-HT6 receptor. After receiving a single oral dose of SAX-187, metabolites M2, M7 and M12 were observed in the plasma of humans. This study was conducted to generate, isolate and characterize metabolites M2, M7 and M12 so that these metabolites could be synthesized. SAX-187 was incubated in rat liver cytosol with acetyl coenzyme A and an acetyl coenzyme A generating system to generate a sufficient amount of M2 for NMR spectroscopic analysis. M7 was produced in rats after receiving a single 100 mg/kg oral dose of M2 and was isolated from the plasma. M12 was generated by incubation of 50 mM SAX-187 with monkey liver microsomes and bicarbonate buffer (pH 8.5) in the presence of UDPGA under a CO2-rich atmosphere. Metabolites M2, M7 and M12 were extracted and isolated using HPLC methods. The purified metabolites were...
SCA-136 is a potent 5 HT2C agonist with an atypical antipsychotic profile that is effective in se... more SCA-136 is a potent 5 HT2C agonist with an atypical antipsychotic profile that is effective in several animal models predictive of antipsychotic activity. A carbamoyl glucuronide (CG) and oxidative metabolites of SCA-136 were formed in liver microsomes of mouse, rat, dog, monkey and human, in bicarbonate buffer and a CO2-enriched environment. CG was the major metabolite in dog and human liver microsomes, while oxidative metabolites were the major metabolites in rat liver microsomes. In human hepatocytes, CG was also the major metabolite. For rats, CG was not detected in plasma or urine, although CG was the major metabolite in rat bile following a single oral 5 mg/kg dose. For dogs, CG was detected in plasma and urine following a single 15 mg/kg oral dose. In mice administered a single oral 50 mg/kg dose, CG was observed in plasma, although not in urine. For monkey, CG was a major metabolite in plasma and urine following a single 25 mg/kg oral dose. CG was one of the major metabolite...
Bazedoxifene (BZA) is a selective estrogen receptor modulator that is being developed for the pre... more Bazedoxifene (BZA) is a selective estrogen receptor modulator that is being developed for the prevention and treatment of postmenopausal osteoporosis. Metabolite profiles of [14C]BZA were investigated in mouse, rat and monkey liver microsomes, as well as in mice, rats and monkeys following a single oral administration of [14C]BZA. In vitro incubations with 50 µM [14C]BZA in the presence of mouse liver microsomes and NADPH produced BZA-N-oxide. CYP450 mediated metabolism of BZA was negligible in rat and monkey liver microsomes. When UDPGA was included in the incubations, BZA was extensively metabolized in all species examined. BZA-4'-glucuronide and BZA-5-glucuronide were observed in all species, while BZA-N-oxide, BZA-4'-glucuronide N-oxide, and BZA-5-glucuronide N-oxide were also generated in mouse liver microsomes. Mouse liver microsomes generated a 2-fold greater amount of the 4'-glucuronide than the 5-glucuronide. Rat liver microsomes generated a 3-fold greater amoun...
Tanaproget (TNPR) is a non-steroidal progesterone receptor agonist which is being investigated fo... more Tanaproget (TNPR) is a non-steroidal progesterone receptor agonist which is being investigated for use as a contraceptive agent. In this study, the in vitro metabolite profiles of TNPR were determined in male and female rat, female dog and human liver microsomes. 14C-TNPR (40 M) was incubated with various liver microsomes (1 mg/mL) in the presence of NADPH and UDPGA in 0.1 M potassium phosphate buffer (pH 7.4) at 37 C for 45 min. Four metabolites were characterized in rats, dogs, and humans; a glucuronide of TNPR (M1), TNPR-N-oxide (M2), a glucuronide of a hydroxy TNPR (M3), and TNPR carbamate (M4). M1 was a major metabolite in all species, while M4 was a predominant metabolite in dogs. Metabolites M2 and M3 were primarily generated in male rat liver microsomes although they were also detected in trace amounts in female rats, dogs, and humans. Metabolites of M1 and M2 were isolated from rat liver microsomal incubations and their structures were characterized by a combination of chro...
In vitro metabolism of [14C]vabicaserin, a potent 5‑HT2C agonist, was evaluated in human liver mi... more In vitro metabolism of [14C]vabicaserin, a potent 5‑HT2C agonist, was evaluated in human liver microsomes and human hepatocytes, and the metabolites were characterized by LC/MS and NMR spectroscopy. Human enzymes involved in vabicaserin metabolism were also identified using selective cytochrome P450 (CYP) chemical inhibitors, commercially available recombinant human CYP enzymes, flavin-containing monooxygenases (FMO), monoamine oxidases (MAO), and UDPGA-glucuronosyltransferases (UGT). In the presence of an NADPH regenerating system, vabicaserin was metabolized by human liver microsomes to four hydroxy metabolites (P1-P4), a nitrone (P5) and an imine (P6). Formation of P6 required microsomes, but not NADPH. When [14C]vabicaserin was incubated with human liver microsomes in bicarbonate buffer and a CO2-enriched environment in the presence of both the NADPH regenerating system and UDPGA, a carbamoyl glucuronide (P7) was observed as the major metabolite, along with the aforementioned me...
Lee/Integrated Strategies for Drug Discovery Using Mass Spectrometry, 2005
... JIM WANG, JACK WANG, MARGARET DAVIS, WILLIAM DEMAIO, JOANN SCATINA, AND RASMY TALAAT ... make... more ... JIM WANG, JACK WANG, MARGARET DAVIS, WILLIAM DEMAIO, JOANN SCATINA, AND RASMY TALAAT ... makes use of extensive expertise in certain technolo-gies, in particular, high-performance liquid-chromatographic (HPLC)mass spectrometry (LC-MS) and HPLC ...
Bazedoxifene (BZA) acetate, a novel estrogen receptor modulator being developed for the preventio... more Bazedoxifene (BZA) acetate, a novel estrogen receptor modulator being developed for the prevention and treatment of post-menopausal osteoporosis, undergoes extensive metabolism in women following oral administration. In this study, the in vitro metabolism of [ 14 C]BZA was determined in human hepatocytes and hepatic and intestinal microsomes, and the UGT isozymes involved in the glucuronidation of BZA were identified. In addition, BZA was evaluated for its potential as a substrate of Pglycoprotein (P-gp) transporter in Caco-2 cell monolayers. BZA was metabolized to two mono-glucuronides, BZA-4'-glucuronide and BZA-5-glucuronide in hepatocytes and in liver and intestinal microsomes including jejunum, duodenum and ileum. Both BZA-4'glucuronide and BZA-5-glucuronide were major metabolites in the intestinal microsomes, while BZA-4'-glucuronide was the predominant metabolite in liver microsomes and hepatocytes. The kinetic parameters of BZA-4'-glucuronide formation were determined in liver, duodenum, and jejunum microsomes and with UGT 1A1, 1A8, 1A10, the most active UGT isoforms involved in the glucuronidation of BZA, while those of BZA-5-glucuronide were determined with all the enzyme systems except in liver microsomes and in UGT 1A1 as the formation of the BZA-5-glucuronide was too low. K m values in liver, duodenum, and jejunum microsomes and UGT 1A1, 1A8, 1A10, were similar and ranged from 5.1 to 33.1 µM for BZA-4'-glucuronide formation and 2.5 to 11.1 µM for BZA-5-glucuronide formation. V max values ranged from 0.8 to 2.9 nmol/min/mg protein for BZA-4'-glucuronide and from 0.1 to 1.2 nmol/min/mg protein for BZA-5-glucuronide. In Caco-2 cells, BZA appeared to be a P-gp substrate.
dm d.aspetjournals.org D ow nloaded from DMD #31179
PloS one, 2016
Parkinson's disease (PD) is a neurodegenerative aging disorder in which postmortem PD brain e... more Parkinson's disease (PD) is a neurodegenerative aging disorder in which postmortem PD brain exhibits neuroinflammation, as well as synucleinopathy-associated protein phosphatase 2A (PP2A) enzymatic activity loss. Based on our translational research, we began evaluating the PD-repurposing-potential of an anti-inflammatory, neuroprotective, and PP2A stimulatory oral drug that is FDA-approved for multiple sclerosis, FTY720 (fingolimod, Gilenya®). We also designed two new FTY720 analogues, FTY720-C2 and FTY720-Mitoxy, with modifications that affect drug potency and mitochondrial localization, respectively. Herein, we describe the metabolic stability and metabolic profiling of FTY720-C2 and FTY720-Mitoxy in liver microsomes and hepatocytes. Using mouse, rat, dog, monkey, and human liver microsomes the intrinsic clearance of FTY720-C2 was 22.5, 79.5, 6.0, 20.2 and 18.3 μL/min/mg; and for FTY720-Mitoxy was 1.8, 7.8, 1.4, 135.0 and 17.5 μL/min/mg, respectively. In hepatocytes, both FTY7...
Desvenlafaxine succinate (DVS) is an orally active serotonin and norepinephrine reuptake inhibito... more Desvenlafaxine succinate (DVS) is an orally active serotonin and norepinephrine reuptake inhibitor in development for treatment of major depressive disorder and vasomotor symptoms. Desvenlafaxine (DV) is a racemic mixture of the major active metabolite of venlafaxine. The metabolite profiles of 14C-DVS were investigated in plasma (1, 4 and 8 hr post-dose) and excreta (up to 24 hr) collected from laboratory animals (mice, rats and dogs) following a single oral administration. In plasma samples collected at 1 and 4 hr post-dose, parent compound represented ≤12% of the radioactivity in mice and rats, and ≤22% of the radioactivity in dogs. For all species, parent compound was not detected by radiochromatography in plasma samples collected at 8 hr post-dose. The major metabolite in plasma from each species was the O-glucuronide conjugate of DV, which represented greater than 76% of the circulating radioactivity. Additional minor oxidative and N-desmethyl metabolites represented less than...
SAX-187 (2-{1-[(6-chloroimidazo[2,1-b][1,3]thiazol-5-yl)sulfonyl]-1H-indol-3-yl}ethanamine hydroc... more SAX-187 (2-{1-[(6-chloroimidazo[2,1-b][1,3]thiazol-5-yl)sulfonyl]-1H-indol-3-yl}ethanamine hydrochloride) is a novel chemical compound with selective agonist activity at the 5-HT6 receptor. After receiving a single oral dose of SAX-187, metabolites M2, M7 and M12 were observed in the plasma of humans. This study was conducted to generate, isolate and characterize metabolites M2, M7 and M12 so that these metabolites could be synthesized. SAX-187 was incubated in rat liver cytosol with acetyl coenzyme A and an acetyl coenzyme A generating system to generate a sufficient amount of M2 for NMR spectroscopic analysis. M7 was produced in rats after receiving a single 100 mg/kg oral dose of M2 and was isolated from the plasma. M12 was generated by incubation of 50 mM SAX-187 with monkey liver microsomes and bicarbonate buffer (pH 8.5) in the presence of UDPGA under a CO2-rich atmosphere. Metabolites M2, M7 and M12 were extracted and isolated using HPLC methods. The purified metabolites were...
SCA-136 is a potent 5 HT2C agonist with an atypical antipsychotic profile that is effective in se... more SCA-136 is a potent 5 HT2C agonist with an atypical antipsychotic profile that is effective in several animal models predictive of antipsychotic activity. A carbamoyl glucuronide (CG) and oxidative metabolites of SCA-136 were formed in liver microsomes of mouse, rat, dog, monkey and human, in bicarbonate buffer and a CO2-enriched environment. CG was the major metabolite in dog and human liver microsomes, while oxidative metabolites were the major metabolites in rat liver microsomes. In human hepatocytes, CG was also the major metabolite. For rats, CG was not detected in plasma or urine, although CG was the major metabolite in rat bile following a single oral 5 mg/kg dose. For dogs, CG was detected in plasma and urine following a single 15 mg/kg oral dose. In mice administered a single oral 50 mg/kg dose, CG was observed in plasma, although not in urine. For monkey, CG was a major metabolite in plasma and urine following a single 25 mg/kg oral dose. CG was one of the major metabolite...
Bazedoxifene (BZA) is a selective estrogen receptor modulator that is being developed for the pre... more Bazedoxifene (BZA) is a selective estrogen receptor modulator that is being developed for the prevention and treatment of postmenopausal osteoporosis. Metabolite profiles of [14C]BZA were investigated in mouse, rat and monkey liver microsomes, as well as in mice, rats and monkeys following a single oral administration of [14C]BZA. In vitro incubations with 50 µM [14C]BZA in the presence of mouse liver microsomes and NADPH produced BZA-N-oxide. CYP450 mediated metabolism of BZA was negligible in rat and monkey liver microsomes. When UDPGA was included in the incubations, BZA was extensively metabolized in all species examined. BZA-4'-glucuronide and BZA-5-glucuronide were observed in all species, while BZA-N-oxide, BZA-4'-glucuronide N-oxide, and BZA-5-glucuronide N-oxide were also generated in mouse liver microsomes. Mouse liver microsomes generated a 2-fold greater amount of the 4'-glucuronide than the 5-glucuronide. Rat liver microsomes generated a 3-fold greater amoun...
Tanaproget (TNPR) is a non-steroidal progesterone receptor agonist which is being investigated fo... more Tanaproget (TNPR) is a non-steroidal progesterone receptor agonist which is being investigated for use as a contraceptive agent. In this study, the in vitro metabolite profiles of TNPR were determined in male and female rat, female dog and human liver microsomes. 14C-TNPR (40 M) was incubated with various liver microsomes (1 mg/mL) in the presence of NADPH and UDPGA in 0.1 M potassium phosphate buffer (pH 7.4) at 37 C for 45 min. Four metabolites were characterized in rats, dogs, and humans; a glucuronide of TNPR (M1), TNPR-N-oxide (M2), a glucuronide of a hydroxy TNPR (M3), and TNPR carbamate (M4). M1 was a major metabolite in all species, while M4 was a predominant metabolite in dogs. Metabolites M2 and M3 were primarily generated in male rat liver microsomes although they were also detected in trace amounts in female rats, dogs, and humans. Metabolites of M1 and M2 were isolated from rat liver microsomal incubations and their structures were characterized by a combination of chro...
In vitro metabolism of [14C]vabicaserin, a potent 5‑HT2C agonist, was evaluated in human liver mi... more In vitro metabolism of [14C]vabicaserin, a potent 5‑HT2C agonist, was evaluated in human liver microsomes and human hepatocytes, and the metabolites were characterized by LC/MS and NMR spectroscopy. Human enzymes involved in vabicaserin metabolism were also identified using selective cytochrome P450 (CYP) chemical inhibitors, commercially available recombinant human CYP enzymes, flavin-containing monooxygenases (FMO), monoamine oxidases (MAO), and UDPGA-glucuronosyltransferases (UGT). In the presence of an NADPH regenerating system, vabicaserin was metabolized by human liver microsomes to four hydroxy metabolites (P1-P4), a nitrone (P5) and an imine (P6). Formation of P6 required microsomes, but not NADPH. When [14C]vabicaserin was incubated with human liver microsomes in bicarbonate buffer and a CO2-enriched environment in the presence of both the NADPH regenerating system and UDPGA, a carbamoyl glucuronide (P7) was observed as the major metabolite, along with the aforementioned me...
Lee/Integrated Strategies for Drug Discovery Using Mass Spectrometry, 2005
... JIM WANG, JACK WANG, MARGARET DAVIS, WILLIAM DEMAIO, JOANN SCATINA, AND RASMY TALAAT ... make... more ... JIM WANG, JACK WANG, MARGARET DAVIS, WILLIAM DEMAIO, JOANN SCATINA, AND RASMY TALAAT ... makes use of extensive expertise in certain technolo-gies, in particular, high-performance liquid-chromatographic (HPLC)mass spectrometry (LC-MS) and HPLC ...