Methylphenidate is stereoselectively hydrolyzed by human carboxylesterase CES1A1

Zejin Sun, Daryl J. Murry, Sonal P. Sanghani, Wilhelmina I. Davis, Natalia Y. Kedishvili, Qin Zou, Thomas Hurley, William F. Bosron

Research output: Contribution to journalArticle

140 Citations (Scopus)

Abstract

Methylphenidate is an important stimulant prescribed to treat attention-deficit hyperactivity disorder. It has two chiral centers, but most current commercial formulations consist of the racemic mixture of the threo pair of methylphenidate isomers (d-, l-threo-methylphenidate). The d-isomer is the pharmacologically active component. Numerous studies reported that oral administration of the methylphenidate racemate undergoes first-pass, stereoselective clearance in humans with l-methylphenidate being eliminated faster than d-methylphenidate. Accordingly, the kinetics of hydrolysis of individual enantiomers by purified native and recombinant human liver carboxylesterases CES1A1 and CES2 and a colon isoenzyme CES3 were examined with a liquid chromatography/mass spectrometry assay. The expression of CES1A1, CES2, and CES3 in Sf9 cells and the methods for purification of the three isoenzymes are reported. CES1A1 has a high catalytic efficiency for both d- and l-enantiomers of methylphenidate. No catalytic activity was detected with CES2 and CES3 for either enantiomer. The catalytic efficiency of CES1A1 for l-methylphenidate (kcat/Km = 7.7 mM-1 min -1) is greater than that of d-methylphenidate (kcat/K m = 1.3-2.1 mM-1 min-1). Hence, the catalytic efficiency of CES1A1 for methylphenidate enantiomers agrees with stereoselective clearance of methylphenidate reported in human subjects. Both enantiomers of methylphenidate can be fit into the three-dimensional model of CES1A1 to form productive complexes in the active site. We conclude that CES1A1 is the major enzyme responsible for the first-pass, stereoselective metabolism of methylphenidate.

Original languageEnglish
Pages (from-to)469-476
Number of pages8
JournalJournal of Pharmacology and Experimental Therapeutics
Volume310
Issue number2
DOIs
StatePublished - Aug 2004

Fingerprint

Carboxylesterase
Methylphenidate
Isoenzymes
Carboxylic Ester Hydrolases
Sf9 Cells
Attention Deficit Disorder with Hyperactivity
Liquid Chromatography
Oral Administration
Mass Spectrometry
Catalytic Domain
Colon

ASJC Scopus subject areas

  • Pharmacology

Cite this

Sun, Z., Murry, D. J., Sanghani, S. P., Davis, W. I., Kedishvili, N. Y., Zou, Q., ... Bosron, W. F. (2004). Methylphenidate is stereoselectively hydrolyzed by human carboxylesterase CES1A1. Journal of Pharmacology and Experimental Therapeutics, 310(2), 469-476. https://doi.org/10.1124/jpet.104.067116

Methylphenidate is stereoselectively hydrolyzed by human carboxylesterase CES1A1. / Sun, Zejin; Murry, Daryl J.; Sanghani, Sonal P.; Davis, Wilhelmina I.; Kedishvili, Natalia Y.; Zou, Qin; Hurley, Thomas; Bosron, William F.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 310, No. 2, 08.2004, p. 469-476.

Research output: Contribution to journalArticle

Sun, Z, Murry, DJ, Sanghani, SP, Davis, WI, Kedishvili, NY, Zou, Q, Hurley, T & Bosron, WF 2004, 'Methylphenidate is stereoselectively hydrolyzed by human carboxylesterase CES1A1', Journal of Pharmacology and Experimental Therapeutics, vol. 310, no. 2, pp. 469-476. https://doi.org/10.1124/jpet.104.067116
Sun, Zejin ; Murry, Daryl J. ; Sanghani, Sonal P. ; Davis, Wilhelmina I. ; Kedishvili, Natalia Y. ; Zou, Qin ; Hurley, Thomas ; Bosron, William F. / Methylphenidate is stereoselectively hydrolyzed by human carboxylesterase CES1A1. In: Journal of Pharmacology and Experimental Therapeutics. 2004 ; Vol. 310, No. 2. pp. 469-476.
@article{5fc65bcbf5594094b5249e3d03bdaddc,
title = "Methylphenidate is stereoselectively hydrolyzed by human carboxylesterase CES1A1",
abstract = "Methylphenidate is an important stimulant prescribed to treat attention-deficit hyperactivity disorder. It has two chiral centers, but most current commercial formulations consist of the racemic mixture of the threo pair of methylphenidate isomers (d-, l-threo-methylphenidate). The d-isomer is the pharmacologically active component. Numerous studies reported that oral administration of the methylphenidate racemate undergoes first-pass, stereoselective clearance in humans with l-methylphenidate being eliminated faster than d-methylphenidate. Accordingly, the kinetics of hydrolysis of individual enantiomers by purified native and recombinant human liver carboxylesterases CES1A1 and CES2 and a colon isoenzyme CES3 were examined with a liquid chromatography/mass spectrometry assay. The expression of CES1A1, CES2, and CES3 in Sf9 cells and the methods for purification of the three isoenzymes are reported. CES1A1 has a high catalytic efficiency for both d- and l-enantiomers of methylphenidate. No catalytic activity was detected with CES2 and CES3 for either enantiomer. The catalytic efficiency of CES1A1 for l-methylphenidate (kcat/Km = 7.7 mM-1 min -1) is greater than that of d-methylphenidate (kcat/K m = 1.3-2.1 mM-1 min-1). Hence, the catalytic efficiency of CES1A1 for methylphenidate enantiomers agrees with stereoselective clearance of methylphenidate reported in human subjects. Both enantiomers of methylphenidate can be fit into the three-dimensional model of CES1A1 to form productive complexes in the active site. We conclude that CES1A1 is the major enzyme responsible for the first-pass, stereoselective metabolism of methylphenidate.",
author = "Zejin Sun and Murry, {Daryl J.} and Sanghani, {Sonal P.} and Davis, {Wilhelmina I.} and Kedishvili, {Natalia Y.} and Qin Zou and Thomas Hurley and Bosron, {William F.}",
year = "2004",
month = "8",
doi = "10.1124/jpet.104.067116",
language = "English",
volume = "310",
pages = "469--476",
journal = "Journal of Pharmacology and Experimental Therapeutics",
issn = "0022-3565",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "2",

}

TY - JOUR

T1 - Methylphenidate is stereoselectively hydrolyzed by human carboxylesterase CES1A1

AU - Sun, Zejin

AU - Murry, Daryl J.

AU - Sanghani, Sonal P.

AU - Davis, Wilhelmina I.

AU - Kedishvili, Natalia Y.

AU - Zou, Qin

AU - Hurley, Thomas

AU - Bosron, William F.

PY - 2004/8

Y1 - 2004/8

N2 - Methylphenidate is an important stimulant prescribed to treat attention-deficit hyperactivity disorder. It has two chiral centers, but most current commercial formulations consist of the racemic mixture of the threo pair of methylphenidate isomers (d-, l-threo-methylphenidate). The d-isomer is the pharmacologically active component. Numerous studies reported that oral administration of the methylphenidate racemate undergoes first-pass, stereoselective clearance in humans with l-methylphenidate being eliminated faster than d-methylphenidate. Accordingly, the kinetics of hydrolysis of individual enantiomers by purified native and recombinant human liver carboxylesterases CES1A1 and CES2 and a colon isoenzyme CES3 were examined with a liquid chromatography/mass spectrometry assay. The expression of CES1A1, CES2, and CES3 in Sf9 cells and the methods for purification of the three isoenzymes are reported. CES1A1 has a high catalytic efficiency for both d- and l-enantiomers of methylphenidate. No catalytic activity was detected with CES2 and CES3 for either enantiomer. The catalytic efficiency of CES1A1 for l-methylphenidate (kcat/Km = 7.7 mM-1 min -1) is greater than that of d-methylphenidate (kcat/K m = 1.3-2.1 mM-1 min-1). Hence, the catalytic efficiency of CES1A1 for methylphenidate enantiomers agrees with stereoselective clearance of methylphenidate reported in human subjects. Both enantiomers of methylphenidate can be fit into the three-dimensional model of CES1A1 to form productive complexes in the active site. We conclude that CES1A1 is the major enzyme responsible for the first-pass, stereoselective metabolism of methylphenidate.

AB - Methylphenidate is an important stimulant prescribed to treat attention-deficit hyperactivity disorder. It has two chiral centers, but most current commercial formulations consist of the racemic mixture of the threo pair of methylphenidate isomers (d-, l-threo-methylphenidate). The d-isomer is the pharmacologically active component. Numerous studies reported that oral administration of the methylphenidate racemate undergoes first-pass, stereoselective clearance in humans with l-methylphenidate being eliminated faster than d-methylphenidate. Accordingly, the kinetics of hydrolysis of individual enantiomers by purified native and recombinant human liver carboxylesterases CES1A1 and CES2 and a colon isoenzyme CES3 were examined with a liquid chromatography/mass spectrometry assay. The expression of CES1A1, CES2, and CES3 in Sf9 cells and the methods for purification of the three isoenzymes are reported. CES1A1 has a high catalytic efficiency for both d- and l-enantiomers of methylphenidate. No catalytic activity was detected with CES2 and CES3 for either enantiomer. The catalytic efficiency of CES1A1 for l-methylphenidate (kcat/Km = 7.7 mM-1 min -1) is greater than that of d-methylphenidate (kcat/K m = 1.3-2.1 mM-1 min-1). Hence, the catalytic efficiency of CES1A1 for methylphenidate enantiomers agrees with stereoselective clearance of methylphenidate reported in human subjects. Both enantiomers of methylphenidate can be fit into the three-dimensional model of CES1A1 to form productive complexes in the active site. We conclude that CES1A1 is the major enzyme responsible for the first-pass, stereoselective metabolism of methylphenidate.

UR - http://www.scopus.com/inward/record.url?scp=3342939961&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3342939961&partnerID=8YFLogxK

U2 - 10.1124/jpet.104.067116

DO - 10.1124/jpet.104.067116

M3 - Article

C2 - 15082749

AN - SCOPUS:3342939961

VL - 310

SP - 469

EP - 476

JO - Journal of Pharmacology and Experimental Therapeutics

JF - Journal of Pharmacology and Experimental Therapeutics

SN - 0022-3565

IS - 2

ER -