Transl Clin Pharmacol.  2016 Sep;24(3):147-151. 10.12793/tcp.2016.24.3.147.

Development and validation of a HPLC-UV method for 4-(4-chlorophenyl)-4-hydroxypiperidine (CPHP), a toxic metabolite of haloperidol, in humans: providing in vivo evidence of CYP3A4-mediated CPHP formation

• Park JY
• Shin JG

• KMID: 2351802
• DOI: http://doi.org/10.12793/tcp.2016.24.3.147

Abstract

We developed a high-performance liquid chromatographic procedure for the determination of 4-(4-chlorophenyl)-4-hydroxypiperidine (CPHP), a toxic metabolite of haloperidol, in human. Chromatographic analysis was performed on a reverse-phase C₁₈ column with a mobile phase containing 50 mM potassium phosphate buffer/acetonitrile (75:25, vol/vol) using UV detection with a wavelength of 220 nm. The limits of detection for CPHP were 1 ng/ml in urine and the assay was linear over the concentration range of 2-500 ng/ml for urine. This analytical method was applied to measure CPHP in human. Nineteen healthy subjects were enrolled and all subjects received a single oral dose of 5 mg haloperidol following a treatment of placebo or itraconazole at 200 mg/day for 10 days in a randomized crossover manner. CPHP was detected in urine samples and average recovered amount of CPHP was 81.31 µg/24 hr in the placebo phase and it was significantly reduced to 30.34 µg/24 hr after itraconazole treatment. The finding provides in vivo evidence that CPHP is an in vivo metabolite of haloperidol in human and its formation is mediated by CYP3A4.

Keyword

Haloperidol; CPHP; CYP3A4; HPLC

MeSH Terms

Chromatography, High Pressure Liquid
Cytochrome P-450 CYP3A
Haloperidol*
Healthy Volunteers
Humans*
Itraconazole
Limit of Detection
Methods*
Potassium
Cytochrome P-450 CYP3A
Haloperidol
Itraconazole
Potassium

Figure

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  Figure 1. Main metabolic pathways of haloperidol.

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  Figure 2. Representative chromatogram of urine samples spiked with 100 ng/ml of CPHP and internal standard (A) of blank urine samples, no spiked blank urine samples (B), and collected urine sample from a healthy subject who administered with 5 mg of haloperidol (C). The labeled chromatographic peaks indicate CPHP (I) and the internal standard metoprolol (II), respectively.

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  Figure 3. Comparison of urinary CPHP recovery in the placebo and itraconazole phases after a single oral administration of 5 mg haloperidol in 19 healthy male subjects.

Reference

1.Kudo S., Ishizaki T. Pharmacokinetics of haloperidol: an update. Clin Pharmacokinet. 1999. 37:435–456.

2.Usuki E., Van der Schyf CJ., Castagnoli N Jr. Metabolism of haloperidol and its tetrahydropyridine dehydration product HPTP. Drug Metab Rev. 1998. 30:809–826.

3.Shin JG., Kane K., Flockhart DA. Potent inhibition of CYP2D6 by haloperidol metabolites: stereoselective inhibition by reduced haloperidol. Br J Clin Pharmacol. 2001. 51:45–52.

4.Fang J., Yu PH., Gorrod JW., Boulton AA. Inhibition of monoamine oxidases by haloperidol and its metabolites: pharmacological implications for the chemotherapy of schizophrenia. Psychopharmacology (Berl). 1995. 118:206–212.

5.Rollema H., Skolnik M., D'Engelbronner J., Igarashi K., Usuki E., Castagnoli N Jr. MPP(+)-like neurotoxicity of a pyridinium metabolite derived from haloperidol: in vivo microdialysis and in vitro mitochondrial studies. J Pharmacol Exp Ther. 1994. 268:380–387.

6.Wright AM., Bempong J., Kirby ML., Barlow RL., Bloomquist JR. Effects of haloperidol metabolites on neurotransmitter uptake and release: possible role in neurotoxicity and tardive dyskinesia. Brain Res. 1998. 788:215–222.

7.Igarashi K., Kasuya F., Fukui M., Usuki E., Castagnoli N Jr. Studies on the metabolism of haloperidol (HP): the role of CYP3A in the production of the neurotoxic pyridinium metabolite HPP+ found in rat brain following ip administration of HP. Life Sci. 1995. 57:2439–2446.

8.Petzer JP., Bergh JJ., Mienie LJ., Castagnoli N Jr.., Van der Schyf CJ. Metabolic defects caused by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and by HPTP (the tetrahydropyridinyl analog of haloperidol), in rats. Life Sci. 2000. 66:1949–1954.

9.Pan LP., Wijnant P., De Vriendt C., Rosseel MT., Belpaire FM. Characterization of the cytochrome P450 isoenzymes involved in the in vitro N-dealkylation of haloperidol. Br J Clin Pharmacol. 1997. 44:557–564.

10.Bowen WD., Moses EL., Tolentino PJ., Walker JM. Metabolites of haloperidol display preferential activity at sigma receptors compared to dopamine D-2 receptors. Eur J Pharmacol. 1990. 177:111–118.

11.Fang J., Yu PH. Effect of haloperidol and its metabolites on dopamine and noradrenaline uptake in rat brain slices. Psychopharmacology (Berl). 1995. 121:379–384.

12.Fukuoka T., Nakano M., Kohda A., Okuno Y., Matsuo M. The common marmoset (Callithrix jacchus) as a model for neuroleptic-induced acute dystonia. Pharmacol Biochem Behav. 1997. 58:947–953.

13.Fornai F., Schlüter OM., Lenzi P., Gesi M., Ruffoli R., Ferrucci M, et al. Parkinson-like syndrome induced by continuous MPTP infusion: convergent roles of the ubiquitin-proteasome system and alpha-synuclein. Proc Natl Acad Sci USA. 2005. 102:3413–3418.

14.Arinobu T., Hattori H., Iwai M., Ishii A., Kumazawa T., Suzuki O, et al. Liquid chromatographic-mass spectrometric determination of haloperidol and its metabolites in human plasma and urine. J Chromatogr B Analyt Technol Biomed Life Sci. 2002. 776:107–113.

15.Fang J., Baker GB., Coutts RT. Determination of 4-(4-chlorophenyl)-4-hy-droxypiperidine, a metabolite of haloperidol, by gas chromatography with electron-capture detection. J Chromatogr B Biomed Appl. 1996. 682:283–288.

16.Higashi Y., Nakamura S., Fujii Y. Sensitive determination of 4-(4-chlorophenyl)-4-hydroxypiperidine, a metabolite of haloperidol, in a rat biological sample by HPLC with fluorescence detection after pre-column derivatization using 4-fluoro-7-nitro-2,1,3-benzoxadiazole. Biomed Chromatogr. 2006. 20:964–970.

17.Park JY., Shon JH., Kim KA., Jung HJ., Shim JC., Yoon YR, et al. Combined effects of itraconazole and CYP2D6∗10 genetic polymorphism on the pharmacokinetics and pharmacodynamics of haloperidol in healthy subjects. J Clin Psychopharmacol. 2006. 26:135–142.

18.Pan LP., De Vriendt C., Belpaire FM. In-vitro characterization of the cytochrome P450 isoenzymes involved in the back oxidation and N-dealkylation of reduced haloperidol. Pharmacogenetics. 1998. 8:383–389.

19.von Moltke LL., Greenblatt DJ., Schmider J., Harmatz JS., Shader RI. Metabolism of drugs by cytochrome P450 3A isoforms. Implications for drug interactions in psychopharmacology. Clin Pharmacokinet. 1995. 29:S33–S43. discussion 43-4.