Skip to main content
NCBI home page
Springer logoLink to Springer
. 2026 May 8;78(3):762–774. doi: 10.1007/s43440-026-00859-1

The novel third-generation antipsychotic drug brexpiprazole affects hepatic transcription factors, serum hormone levels, and cytochrome P450 enzymes after prolonged treatment: pharmacological significance

Przemysław J Danek 1, Renata Pukło 1, Władysława A Daniel 1,✉
PMCID: PMC13275521  PMID: 42101808

Abstract

Background

Brexpiprazole is a third-generation antipsychotic used for the treatment of schizophrenia or as an adjunctive drug for the treatment of affective and neurological disorders. The combined treatments with brexpiprazole and other drugs that are also cytochrome P450 (CYP) substrates may lead to pharmacokinetic drug-drug interactions. Our present work aimed to investigate the effects of prolonged administration of brexpiprazole on the expression of hepatic transcription factors and CYP drug-metabolizing enzymes.

Methods

Male Wistar rats received brexpiprazole (1 mg/kg ip.) for two weeks. Their livers were excised 24 h after the last dose, and the activities (HPLC), protein levels (Western blotting), and mRNAs (qRT-PCR) of CYP enzymes were measured. In parallel, the expression of hepatic transcription factors (Western blotting, qRT-PCR) and the concentration of serum hormones (ELISA) were assessed.

Results

Brexpiprazole produced a broad-spectrum effect on CYP expression and activity. It enhanced the expression and activity of CYP1A, CYP2A, CYP3A1/2, and the activity of CYP2B, but decreased the expression/activity of CYP2Ds and CYP2E1. The observed changes in CYP enzymes corresponded to alterations in transcription factors: the increased expression of PXR and AhR, and decreased expression of PPARγ, LXR, and FXR. The above modifications in CYP enzymes’ expression were accompanied by enhanced corticosterone and reduced T4 serum levels.

Conclusions

Brexpiprazole affects the expression of hepatic transcription factors and CYP, which may impact its own biotransformation and the metabolism of endogenous substances and concomitantly administered drugs, and lead to drug-drug interactions of pharmacological/clinical importance.

Supplementary information

The online version contains supplementary material available at 10.1007/s43440-026-00859-1.

Keywords: Brexpiprazole, Cytochrome P450 (CYP), Liver, Enzyme expression/activity, Transcription factors, Serum hormones

Introduction

Brexpiprazole is a third-generation antipsychotic drug approved for therapy in 2015. Thanks to its unique receptor profile, the drug is effective in the treatment of both positive and negative symptoms of schizophrenia and shows a low risk of side effects compared with earlier antipsychotics [1–4]. Brexpiprazole is used for the treatment of schizophrenia or as an adjunctive drug for the treatment of depression, posttraumatic stress disorders, and obsessive-compulsive disorders [5–10] and exerts a beneficial effect in the treatment of psychosis and agitation in Alzheimer’s disease [11, 12]. Brexpiprazole affects the brain dopaminergic, noradrenergic, and serotonergic systems, which are involved in those psychiatric diseases. It acts as a partial agonist at serotonin 5-HT1A and dopamine D2/3 receptors, and has a strong antagonistic action at serotonin 5-HT2A and noradrenaline α1B/α2C receptors, which was shown in vitro and confirmed in animal tests in vivo [13, 14].

The pharmacokinetics of brexpiprazole, 7-[4-[4-(1-benzothiophen-4-yl)piperazin-1-yl]butoxy]quinolin-2(1 H)-one, can be described by a two-compartment model. It indicates that the pharmacokinetic processes of this compound occur with varying intensity in individual organs/tissues, resulting in its uneven distribution throughout the body. The drug is highly bound to plasma proteins and distributed widely in the organism, reaching the Vd value around 2 L/kg [15–17]. The main enzymes involved in the metabolism of brexpiprazole are CYP3A4 and CYP2D6 [18–20]. Brexpiprazole is broken down into two metabolites, metabolite DM-3411 (produced by CYP3A4 and CYP2D6) and DM-3412 (primarily formed by CYP2D6) [18]. The major metabolite DM-3411, formed by S-oxidation, shows low affinity for D2 receptor and poorly penetrates the blood-brain barrier (in contrast to other metabolites), so it does not contribute to the pharmacological effect of the antipsychotic. Moreover, brexpiprazole undergoes bioactivation to form reactive intermediate metabolites [21].

Studies to date have not shown any significant effect of the drug on hepatic cytochrome P450 (CYP) in in vitro experimental models. Studies with human liver microsomes showed that brexpiprazole exhibited weak inhibitory effects on CYP2C9, CYP2C19, CYP2D6, and CYP3A4, but had a moderate inhibitory activity on CYP2B6. Moreover, brexpiprazole did not affect the mRNA level of CYP1A2, CYP2C9, and CYP3A4 after 3-day exposure of human hepatocyte culture to therapeutic concentrations of the antipsychotic [15]. However, the effect of prolonged administration of the antipsychotic on the CYP enzymes’ expression in vivo has not been investigated as yet.

The cytochrome P450 families CYP1-3 take part in the oxidative metabolism of drugs, including psychotropics [22, 23], and endogenous substances, such as steroids, fatty acids, or monoaminergic neurotransmitters [24]. The expression of genes coding for CYP enzymes involved in drug metabolism can be transcriptionally activated or hampered by xenobiotics, including drugs, through xenobiotic receptor-dependent mechanisms. Several receptors mediate the induction of these CYP genes, such as the aryl hydrocarbon receptor (AhR), the glucocorticoid receptor (GR), the pregnane X receptor (PXR), the constitutive androstane receptor (CAR), and peroxisome proliferator-activated receptor (PPAR) [25–28]. In addition, farnesoid X receptor (FXR) and liver X receptor (LXR), which mainly regulate CYP enzymes engaged in the metabolism of cholesterol, can directly or indirectly affect the expression of some CYP genes encoding drug-metabolizing enzymes [29–36].

Psychotropic drugs, including antipsychotics, are administered to patients for a long time, and very often in combinations with other clinically important drugs that are substrates of CYP enzymes [37]. Therefore, the inhibition and/or induction of CYP enzymes by those drugs may produce drug-drug interactions of pharmacological and clinical importance. Considering the role of CYP in the metabolism of vital endogenous and exogenous substrates, as well as long-term treatment of schizophrenia, our present work aimed to investigate the effects of prolonged administration of brexpiprazole on the CYP drug-metabolizing enzymes’ expression and activity in the rat liver. Additionally, we have identified the effect of chronic antipsychotic treatment on the expression of transcription factors and on serum levels of hormones regulating hepatic CYP enzymes participating in the metabolism of drugs and endogenous substrates.

Materials and methods

Drugs and chemicals

Brexpiprazole (BREX) was supplied by TargetMol (Boston, MA, USA; T8690). NADP (N0505), NADPH (N7505), glucose-6-phosphate-dehydrogenase (G7877), glucose-6-phosphate (G7879), Tween 80, methylcellulose, caffeine and its metabolites (C8960), bufuralol and its metabolite 1’-hydroxybufuralol (UC168), chlorzoxazone and its metabolite 6-hydroxychlorzoxazone (C4397) were obtained from Sigma (St. Louis, MO, USA). Steraloids (Newport, KY, USA) provided testosterone and its metabolites. Warfarin was obtained from Merck (Darmstadt, Germany), and 7-hydroxywarfarin was synthesized at the Maj Institute of Pharmacology, Kraków, Poland. For Western-blot analysis, the primary rabbit polyclonal anti-rat CYP2C11 (1:2000; PA3-034), CYP2E1 (1:2000; PA5-52652), PXR (1:1000; PA5-116715), PPARγ (1:1000; PA3-821A), FXR (1:2000; PA5-40755), LXR (1:2000; 14351–1-AP) and CAR (1:1000; PA5-36268) antibodies or mouse anti-rat AhR (1:1000; MA1-514) from Thermo Fisher Scientific (Walthman, MA, USA), anti-rat CYP3A1 (1:2000; Ab1253) and CYP3A2 (1:2000; Ab1276) from Millipore (Temecula, USA), anti-human CYP2D6 (1:2000; FNab02157) and CYP2A6 (1:1000; FNab02165) antibodies from FineTest (Wuhan, China), anti-rat β-actin antibody (1:15000; A1978) from Sigma (St. Louis, MO), USA), the monoclonal mouse anti-rat CYP2B1/2B2 (1:2000; SC-53242), CYP2C6 (1:2000; SC-73484), and CYP1A2 (1:2000; SC53241) antibodies from Santa Cruz Biotechnology (Dallas, TX, USA) were used. Horseradish peroxidase-labeled secondary antibodies: goat anti-mouse antibodies (1:2000; PI-1000–1) were obtained from Jackson ImmunoResearch (West Grove, PA, USA), and goat anti-rabbit antibodies (1:2000; 115–005-003) were from Vector Laboratories (Burlingame, CA, USA). Rat cDNA-expressed CYP1A2, CYP2B1, CYP2C6, CYP2C11, CYP2E1, CYP2D1, CYP3A1, CYP3A2, or human CYP2A6 (Supersomes) were from Gentest Corp. (Woburn, MA, USA). The chemiluminescence reagents SuperSignal West Pico PLUS Chemiluminescent Substrate kit (34077) came from Thermo Fisher Scientific (Walthman, MA, USA). For RNA isolation, a Total RNA Mini kit (031–100, A&A Biotechnology, Gdynia, Poland) was applied, while a High-Capacity cDNA Reverse Transcription Kit (4368814), TaqMan assays, and the TaqMan Gene Expression Master Mix (4369016) from Life Technologies (Carlsbad, CA, USA) were used for mRNA estimation. The ELISA kits provided by Bioassay Technology Laboratory (Shanghai, China) were employed to measure serum concentrations of growth hormone (E1997Ra), corticosterone (EA0036Ra), and thyroid hormones (T3, T4) (EA0027Ra, E0853Ra). All the organic solvents of HPLC purity were provided by Merck (Darmstadt, Germany).

Animals

Adult male Wistar Han rats (3 months old and weighing 280–300 g) from Charles River Laboratories (Sulzfeld, Germany) were used in all experiments. The animals were initially acclimatized for 10 days and housed in environmentally controlled rooms (22 ± 2 °C room temperature, 50 ± 5% room humidity, and 12:12 light: dark cycle) with free access to tap water, standard laboratory food, and environmental enrichment provided to each cage (6 animals per cage). During this time, the rats passively habituated to the experimenter’s presence and handling procedures – once a day before the experiment begins. All experiments were conducted in accordance with the NIH Guide for the Care and Use of Laboratory Animals, and the experimental protocols were approved by the Local Ethics Commission for Experimentation on Animals at the Maj Institute of Pharmacology, Polish Academy of Sciences, Kraków (approval code: 198/2018; date: 7.06.2018).

Animal treatment

The rats (n = 15 for each treatment group) received intraperitoneal injections of brexpiprazole (1 mg/kg) or vehicle control (0.5% methylcellulose and 0.2% Tween 80 in sterile water) once a day for two weeks. The solution of drugs was prepared daily. Based on the literature data, it can be assumed that the applied dosage of brexpiprazole resulted in a concentration in rat blood plasma within the range of therapeutic drug concentrations in patients [15, 38, 39]. Moreover, the selected dose of the tested antipsychotic was active in pharmacological and behavioral experimental models [20]. 24 hours after the last dose, the rats were sacrificed by decapitation. The trunk blood was collected immediately and placed in the fridge. The livers were quickly removed, frozen in dry ice, and stored at −80 °C. Microsomes were prepared from individual livers by differential centrifugation, according to the previously used method [40]. Blood serum was obtained through centrifugation (2500 ×g, 15 min) and stored at −80 °C until analysis.

CYP enzyme activity in rat liver microsomes

The activities of CYP enzymes were assessed in liver microsomal suspensions by measuring the transformation rates of specific metabolic substrates and reactions under linear conditions, as reported previously [40, 41]. Briefly, reactions were carried out in an incubation mixture containing rat liver microsomes (ca. 1 mg of protein/ml), an NADPH or NADPH-generating system, and a CYP-specific substrate. The activity of CYP1A was estimated by measuring the rate of caffeine C-8-hydroxylation and 3-N-demethylation (100 μM). The CYP2C6 activity was studied by estimating the rate of warfarin 7-hydroxylation (60 μM). The CYP2D activity was determined using the 1’-hydroxylation of bufuralol (10 μM). The activity of CYP2E1 was studied by measuring the rate of 6-hydroxylation of chlorzoxazone (200 μM). The activities of CYP2A, CYP2B, CYP2C11, and CYP3A were evaluated by measuring the rates of CYP-specific reactions: the 7α-, 16β-, 2α-, and 6β-hydroxylation of testosterone (100 μM), respectively. Specific substrate and its metabolites were analyzed by HPLC with UV or fluorometric detection.

Determination of CYP enzymes’ and transcription factors’ protein levels in rat liver

Western blot was performed as described previously with some modifications [42, 43]. In brief, samples (10 μg of microsomal protein for CYP enzymes or 30 μg of total protein for transcriptional factors) were separated on a 12% or 8% (for AhR factor) SDS-PAGE gel in a Laemmli buffer system, and electrotransferred (100 V, 90 min) onto a nitrocellulose membrane. Then, the membranes were blocked overnight at 4 °C with 5% nonfat milk in Tris-buffered saline with 0.1% Tween 20. The blots were probed with appropriate primary antibodies and subsequently incubated with the corresponding horseradish peroxidase-conjugated secondary antibodies. The bands were evaluated using a luminescent image analyzer (IBright 1500, Thermo Fisher Scientific Walthman, MA, USA) and relative levels of immunoreactivity were quantified using the Image Gauge software (Fuji Film, USA). The measurement results were normalized to the protein level based on the β-actin levels.

Measurement of CYP enzymes’ and transcriptional factors’ mRNAs in the liver

The RT–qPCR method conducted in this study has been previously described by Danek et al. [42, 43]. In brief, the liver tissue was dissected, quickly frozen in dry ice, and stored at − 80 °C until analysis. The frozen liver tissue fragments (20 mg) were homogenized, and the total RNA was extracted using a Total RNA Mini kit. The total RNA concentration was measured using a Synergy/HTX multi-mode reader (BioTek, Winooski, VT, USA). The isolated RNA was then reverse-transcribed using a Transcriptor High-Fidelity cDNA synthesis kit. Quantitative real-time PCR was carried out using TaqMan Gene Expression Assays, TaqMan Expression Master Mix, species-specific TaqMan probes, and primers for the selected genes: CYP1A1 (Rn00487218_m1), CYP1A2 (Rn1422531_m1), CYP2B1 (Rn01457875_m1), CYP2B2 (Rn02786833_m1), CYP2C6 (Rn00569868_m1), CYP2D1 (Rn01775090_mH), CYP2D2 (Rn00562419_m1), CYP2D3 (Rn00597330_m1), CYP2D4 (Rn00593393_m1), CYP2E1 (Rn00580624_m1), CYP2C11 (Rn01502203_m1), CYP3A1 (Rn03062228_m1), CYP3A2 (Rn00756461_m1), Nr1i2 (PXR, Rn00583887_m1) Nr1i3 (CAR, Rn000576085_m1), AhR (Rn00565750_m1), PPARγ (Rn00440945_m1), Nr1h4 (FXR, Rn00572658_m1), Nr1h3 (LXR, Rn00581185_m1), and the reference gene ACTB (Rn00667869_m1), encoding β-actin. Quantitative real-time PCR was performed in duplicate using the BioRad CFX96 PCR system (BioRad, Hercules, CA, USA). Gene expression was determined using the 2-delta Ct method with the β-actin gene (ACTB) expression serving as the reference.

Assessment of hormone levels

The serum concentrations of GH, corticosterone, and thyroid hormones (T3, T4) were measured using ELISA kits. Absorbance was read using a Synergy Mx Monochromator-Based Multi-Mode Microplate Reader (Biotek, Winooski, VT, USA).

Statistical analysis

The statistical significance of drug effect on enzyme activity, protein level, or gene expression was calculated using Student’s t-test compared to the control value (GraphPad Prism 10.0; GraphPad Prism Software Inc., CA, USA). The Kolmogorov–Smirnov test was used to assess normality. The results are reported as the mean ± SEM. The changes produced by brexpiprazole were considered statistically significant when p < 0.05. CYP activity and protein levels were quantified relative to the control group, and results are presented as per cent of control; mRNA levels were quantified relative to the control group, and results are presented as fold change relative to ACTB gene; hormone levels were quantified relative to the control group, and results are presented as per cent of control.

Results

The impact of prolonged administration of brexpiprazole on the CYP activity in the liver

The obtained results indicated significant alterations in the activity of CYP enzymes in rat liver during prolonged treatment with the examined new atypical neuroleptic (Fig. 1A). The activity of CYP1A (measured as the caffeine C-8-hydroxylation), CYP2A (testosterone 7α-hydroxylation), CYP2B (testosterone 16β-hydroxylation), CYP2C11 (testosterone 2α-hydroxylation), CYP3A (testosterone 2β-hydroxylation) significantly increased up to 131% (Student’s t-test; p = 0.0049; t28 = 3.051), 137% (Student’s t-test; p = 0.0086; t25 = 2.854), 134% (Student’s t-test; p = 0.037; t23 = 2.215), 136% (Student’s t-test; p = 0.8423; t28 = 0.2008) and 137% (Student’s t-test; p = 0.016; t28 = 2.547) of the control, respectively, after a two-week administration of brexpiprazole. In contrast, the activity of CYP2D (bufuralol 1′-hydroxylation) and CYP2E1 (chlorzoxazone 6-hydroxylation) was reduced by the examined drug down to 52% (Student’s t-test; p < 0.0001; t26 = 7.459) and 87% (Student’s t-test; p = 0.023; t28 = 2.403) of the control, respectively. The studied neuroleptics did not produce any significant effect on CYP2C6 activity (warfarin 7-hydroxylation) when administered in vivo.

Fig. 1.

Fig. 1

Open in a new tab

The effect of two-week administration of brexpiprazole on the activity (A), protein level (B), and mRNA level (C) of liver CYP enzymes/genes. All values are shown as the mean ± SEM, each data point represents an individual biological sample, CYP activity was quantified relative to the control group and results are presented as per cent of control (n = 15); protein levels were quantified relative to the control group and results are presented as per cent of control (n = 12); mRNA levels were quantified relative to the control group and results are presented as Fold change relative to ACTB gene (n = 8). Statistical significance was assessed by Student’s t-test and indicated as * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, compared to the control. the control values (nmol/mg protein/min) for CYP enzyme activities were: 8.87 ± 2.17 (caffeine 8-hydroxylation); 1.64 ± 0.25 (caffeine 3-N-demethylation); 170.89 ± 20.18, 23.85 ± 9.79, 327.88 ± 156.9, 18.34 ± 5.3, and 218.61 ± 71.63 (7α-, 16β-, 2α-, 2β-, and 6β-hydroxylation of testosterone, respectively); 5.17 ± 1.29 (warfarin 7-hydroxylation), 3.1 ± 0.48 (bufuralol 1-hydroxylation), and 3.3 ± 0.53 (chlorzoxazone 6-hydroxylation). C-8, caffeine 8-hydroxylation; 3-N, caffeine 3-N-demethylation; 7α-, 16β-, 2α-, 2β-, and 6β-hydroxylation of testosterone; WARF 7-OH, warfarin 7-hydroxylation; CHZ 6-OH, chlorzoxazone 6-hydroxylation; BUF 1’-OH, bufuralol 1-hydroxylation. CYP enzymes: 1A subfamily (1A1, 1A2), 2A subfamily (2A1, 2A2), 2B subfamily (2B1, 2B2), 2C11, 3A subfamily (3A1, 3A2), 2C6, 2E1, 2D subfamily (2D1, 2D2, 2D3, 2D4)

The influence of chronic brexpiprazole treatment on the CYP protein level in the liver

Western blot analysis was used to evaluate the protein levels of CYP enzymes in the liver microsomes of the control group and of animals after prolonged treatment with brexpiprazole. In most cases, the observed changes in CYP protein levels corresponded well with the alterations in CYP activities (Figs. 1B, 2, Fig. S1). Brexpiprazole increased the protein level of CYP1A, CYP2A and CYP3A2 up to 142% (Student’s t-test; p = 0.0387; t22 = 2.199), 158% (Student’s t-test; p = 0.0412; t22 = 2.168) and 133% (Student’s t-test; p = 0.0071; t22 = 2.967) of the control, respectively. Such a tendency was also observed for CYP2C11 and CYP3A1. At the same time, the protein level of CYP2E1 and CYP2D decreased to 73% (Student’s t-test; p < 0.0001; t22 = 4.605) and 66% (Student’s t-test; p < 0.0001; t22 = 5.075) of the control, respectively. On the other hand, the protein levels of CYP2B and CYP2C6 were not affected by brexpiprazole.

Fig. 2.

Fig. 2

Open in a new tab

The effect of 2-week treatment with brexpiprazole on the intensity of protein bands of CYP enzymes (CYP1A, CYP2A, CYP2B, CYP2C6, CYP2C11, CYP3A1, CYP3A2, CYP2E1, CYP2D) in rat liver microsomes. Microsomal proteins were subjected to the Western immunoblot analysis. The appropriate cDNA-expressed CYP enzymes were used as standards. The representative CYP protein bands are shown. The presented results are typical of six separate rats per group. The mean values ± SEM (n = 12) are shown in Fig. 1. The bands were evaluated using a luminescent image analyzer (IBright 1500, Thermo Fisher Scientific Walthman, MA, USA) and relative levels of immunoreactivity were quantified using the image gauge software (Fuji Film, USA). STD, standard

The effect of prolonged brexpiprazole treatment on the hepatic CYP mRNA level

The mRNA levels of CYP genes were measured in the liver tissue to find out the molecular mechanism of the observed changes in the activity and protein levels of the investigated CYP enzymes. Most of the observed alterations in CYP protein levels and activity corresponded well with mRNA level (Fig. 1C). Two-week administration of brexpiprazole significantly enhanced the mRNA level of CYP1A1 (Student’s t-test; p = 0.001; t14 = 4.155), CYP1A2 (Student’s t-test; p = 0.034; t14 = 2.368), CYP2A1 (Student’s t-test; p = 0.0157; t14 = 2.748), CYP2A2 (Student’s t-test; p = 0.041; t14 = 2.251), CYP3A1 (Student’s t-test; p = 0.0198; t14 = 2.629) and CYP3A2 (Student’s t-test; p = 0.0269; t14 = 2.471), but diminished that of CYP2D1 (Student’s t-test; p = 0.003; t14 = 3.583), CYP2D2 (Student’s t-test; p = 0.0377; t14 = 2.295), CYP2D3 (Student’s t-test; p = 0.0033; t14 = 3.533), CYP2E1 (Student’s t-test; p = 0.0452; t14 = 2.199), and surprisingly also CYP2C6 (Student’s t-test; p = 0.0482; t14 = 2.164). The studied drug did not affect significantly the mRNA level of CYP2B1, CYP2B2, and CYP2C11.

The effect of two-week brexpiprazole administration on the expression of transcription factors in the liver

In the next step of our study, we focused on finding the molecular mechanism of changes in the expression of the investigated CYP enzymes in the liver, produced by repeated administration of brexpiprazole. For this purpose, we determined the effect of the studied drug on the protein level and mRNA of transcription factors regulating CYP genes.

Brexpiprazole evoked a distinct increase in the protein level of PXR and AhR up to 162% (Student’s t-test; p = 0.0177; t22 = 2.563) and 142% (Student’s t-test; p = 0.0494; t21 = 2.086) of the control, respectively (Figs. 3A, 4, Fig. S2). Moreover, brexpiprazole significantly reduced the protein level of FXR and LXR down to 58% (Student’s t-test; p < 0.0001; t22 = 5.317) and 79% (Student’s t-test; p = 0.0004; t22 = 4.134) of the control, respectively, and such a tendency was also observed for PPARγ. The examined drug did not affect the protein level of the CAR transcription factor in rat liver after chronic treatment.

Fig. 3.

Fig. 3

Open in a new tab

The effect of prolonged treatment with brexpiprazole on the expression of transcription factors in the liver: protein level (A) and mRNA level (B). All values are shown as the mean ± SEM, each data point represents an individual biological sample. Protein levels were quantified relative to the control group and results are presented as per cent of control (n = 12); mRNA levels were quantified relative to the control group and results are presented as Fold change relative to the ACTB gene (n = 8). Statistical significance was assessed by Student’s t-test and indicated as * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, compared to the control. PXR, pregnane X receptor; CAR, constitutive androstane receptor; AhR, aryl hydrocarbon receptor; PPARγ, peroxisome proliferator-activated receptor γ; LXR, liver X receptor; FXR, farnesoid X receptor

Fig. 4.

Fig. 4

Open in a new tab

The effect of 2-week treatment with brexpiprazole on the intensity of protein bands of transcription factors (PXR, CAR, AhR, PPARγ, LXR, FXR) in the rat liver. Whole liver proteins were subjected to the Western immunoblot analysis. A set of protein markers was used as a standard. The representative protein bands are shown. The presented results are typical of six separate rats per group. The mean values ± SEM (n = 12) are shown in Fig. 3. The bands were evaluated using a luminescent image analyzer (IBright 1500, Thermo Fisher Scientific Walthman, MA, USA), and relative levels of immunoreactivity was quantified using the image gauge software (Fuji Film, USA). PXR, pregnane X receptor; CAR, constitutive androstane receptor; AhR, aryl hydrocarbon receptor; PPARγ, peroxisome proliferator-activated receptor γ; LXR, liver X receptor; FXR, farnesoid X receptor

The above changes in the protein levels of the transcription factor correlated positively with the corresponding mRNA levels. Prolonged treatment with brexpiprazole enhanced the mRNA level of PXR (Student’s t-test; p = 0.0317; t14 = 2.386) and AhR (Student’s t-test; p = 0.0387; t14 = 2.347) (Fig. 3B). At the same time, the antipsychotic reduced the mRNA level of PPARγ (Student’s t-test; p = 0.0199; t14 = 2.628), FXR (Student’s t-test; p = 0.0024; t14 = 3.703) and LXR (Student’s t-test; p = 0.0022; t14 = 3.728). The examined antipsychotic did not affect the mRNA level of CAR in the rat liver after two weeks of treatment.

The effect of chronic administration of brexpiprazole on serum hormone concentrations

The serum concentrations of hormones participating in the physiological regulation of drug-metabolizing CYP enzymes have been measured. Prolonged administration of brexpiprazole increased the serum concentration of corticosterone up to 182% (Student’s t-test; p = 0.0491; t10 = 2.239) and diminished that of T4 to 69% (Student’s t-test; p = 0.009; t11 = 3.166) of the control (Fig. 5). The concentration of growth hormone and T3 remained unchanged after brexpiprazole treatment.

Fig. 5.

Fig. 5

Open in a new tab

The effect of prolonged treatment with brexpiprazole on serum hormone concentrations. All values are shown as the mean ± SEM, each data point represents an individual biological sample. Hormone levels were quantified relative to the control group and results are presented as a percentage of control (n = 8). Statistical significance was assessed by Student’s t-test and indicated as * p < 0.05, ** p < 0.01, compared to the control. The absolute control values were: 144.6 ± 4.7 pg/ml, 292 ± 72.5 ng/ml, 2.26 ± 0.13 ng/ml, 66.3 ± 8.4 ng/ml, for growth hormone (GH), corticosterone (CORT), triiodothyronine (T3), and thyroxine (T4), respectively, in blood serum

Discussion

The obtained results evidence significant effects of the tested antipsychotic drug on the CYP expression and activity, as well as on the expression of CYP-regulating transcriptional factors and serum hormones (Table 1). Brexpiprazole enhanced the expression and activity of CYP1A, CYP2A, and CYP3A, but reduced the expression and activity of CYP2E1 and CYP2D. Moreover, it increased the CYP2B activity with no change in the CYP2B1/2 mRNA and protein levels. The observed changes in the studied CYP enzymes mostly corresponded with the simultaneously occurring alterations in the levels of transcription factors and serum hormones. This is the first study showing the effect of prolonged treatment with brexpiprazole on the expression and activity of CYP enzymes in rat liver.

Table 1.

Summary of the effects of two-week administration of brexpiprazole on the hepatic cytochrome P450 enzymes and transcriptional factors, and serum hormone levels

CYP enzymes
CYP mRNA level Protein level Activity
1A1 ↑ ↑ ↑
1A2 ↑
2A1 ↑ ↑ ↑
2A2 ↑
2B1 − − ↑
2B2 −
2C6 ↓ − −
2C11 − − −
2D1 ↓ ↓ ↓
2D2 ↓
2D3 ↓
2D4 −
2E1 ↓ ↓ ↓
3A1 ↑ − ↑
3A2 ↑ ↑
Transcriptional factors
mRNA level Protein level
PXR ↑ ↑
CAR − −
AhR ↑ ↑
PPARγ − ↓
FXR ↓ ↓
LXR ↓ ↓
Hormone levels
Growth hormone −
Corticosterone ↑
Triiodothyronine (T3) −
Thyroxine (T4) ↓
Open in a new tab

↑ increase, ↓ decrease, (↓) tendency to decrease, − no change

It was previously shown that brexpiprazole had some mild effect on certain CYP enzymes in vitro. Sasabe et al. [16] demonstrated that brexpiprazole exhibited a direct, weak inhibitory effect on CYP2B6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 in human liver microsomes. In the same study, brexpiprazole at a high concentration of 10 μM (far exceeding therapeutic concentrations [38]), acting for three days, increased the mRNA level of CYP1A2 and CYP3A4 (but not their activity), and did not affect CYP2C9 and CYP2B6 in the culture of human hepatocytes. Thus, both in human hepatocytes in vitro [16] and in rat liver in vivo (present study), brexpiprazole induced the expression of CYP1A2 and CYP3A genes. However, in our in vivo studies carried out on rats, the effect of brexpiprazole on the enzyme expression was accompanied by enhancement of enzyme activity, and was achieved at therapeutic concentration provided by a low intraperitoneal dose of 1 mg/kg, acting for two weeks. This suggests that brexpiprazole action in the two species shares some molecular mechanisms, which lead to an increase in enzyme expression; however, simultaneous direct enzyme inhibition by brexpiprazole occurring at a high 10 μM drug concentration masks the molecular effect of induction on enzyme activity in the culture of human hepatocytes. On the other hand, prolonged treatment in vivo may trigger additional mechanisms (e.g., neuroendocrine mechanisms) that evoke enhancement in the activity of CYP1A2 and CYP3A enzymes.

Long-term administration of brexpiprazole significantly increased the expression (mRNA and protein levels) of AhR in rat liver, which was consistent with the induction of CYP1A1 and CYP1A2 expression and activity, and with the current knowledge on the mechanism of CYP1A enzyme induction [44]. This may indicate that brexpiprazole indirectly affects CYP1A in the liver (via increased AhR expression), which does not exclude the possibility of a direct agonistic effect of the antipsychotic on AhR. It was shown that β-naphthoflavone, through its effect on AhR, might affect CYP1A induction in various species, including rats and humans [45], which may suggest that brexpiprazole acts in the same way. The observed induction of CYP1A enzymes may act not only on the rate of drug biotransformation (antidepressants, antipsychotics), but also on the metabolism of endogenous substrates, such as steroid hormones (progestogens, estrogens), melatonin, retinol, linoleic acid, and phosphatidylcholines [46].

The observed increase in the expression and activity of CYP3A and CYP2A, involved in the metabolism of most drugs and steroids [22], corresponded well with the rise in the PXR expression, the main transcription factor inducing those CYP genes [27, 47–50]. In addition to the elevated PXR, the enhancement in the expression of CYP3A, detected in the present study, may also be due to the LXR down-regulation by brexpiprazole. It has been shown that the expression of Cyp3a was higher in LXR-deficient mice [30]. On the other hand, FXR was demonstrated to transcriptionally up-regulate the expression of CYP1A1 and CYP3A enzymes in mouse and human cell lines [32, 36], so their down-regulation by brexpiprazole could attenuate the effect of the elevation of AhR and PXR on those CYP genes, respectively. The observed decrease in the expression of FXR and LXR by prolonged brexpiprazole treatment may be of particular physiological importance, since these transcription factors take part in the regulation of CYP enzymes engaged in the metabolism of cholesterol [32].

Interestingly, brexipiprazole down-regulated the expression of PPARγ transcription factor, which might be related to the observed decrease in expression of CYP2D genes (CYP2D1–D3). Zhang et al. demonstrated the involvement of PPARs in the regulation of CYP2D by growth hormone in the brain, with PPARγ activating the CYP2D6 gene promoter while PPARα inhibiting its function [51]. The observed reduction in PPARγ expression may also be important for the expression of CYP4 family enzymes, which are involved in fatty acid metabolism [52]. Moreover, considering that the main enzymes involved in the metabolism of brexpiprazole are CYP3A4 and CYP2D6 [18–20], and the investigated antipsychotic affects these enzymes, the rate of its biotransformation pathways may be changed during prolonged treatment.

It is also worth noting that the transcription factors mentioned above (PXR, CAR, AhR, LXR) are involved not only in regulating the expression of CYP enzymes, but also other proteins important for proper body function. For example, they are involved in regulating the expression of P-glycoprotein, which transports endogenous substrates (cortisol, cholesterol), toxins, and many drugs, including psychotropics [53, 54]. Therefore, changes in their expression levels may have broader physiological and pharmacological implications.

The expression and activity of hepatic CYP enzymes can also be regulated via the neuroendocrine pathways, which remain under control of the central nervous system, including the dopaminergic, noradrenergic, and serotonergic systems [55–58]. By acting with high affinity on serotonin, dopamine, and noradrenaline receptors, brexpiprazole may influence the secretion of hormones regulating the expression of enzymes belonging to the CYP1-3 families [13, 14]. It was shown that corticosterone up-regulated the expression of CYP1A, CYP2B, CYP3A, CYP2C11, and growth hormone did so to the CYP2C11 and CYP3A expression and activity in the rat liver [59]. It seems, therefore, that another possible cause of the brexpiprazole effects on CYP enzymes’ expression may lie in the drug’s ability to interact with different receptors in the brain and, in consequence, in influencing the secretion of CYP-regulating hormones. In our experiment, brexpiprazole increased the serum concentration of corticosterone, which is an agonist of the glucocorticoid receptor (GR), acting directly or indirectly (via increasing PXR and CAR expression) on the CYP3A and CYP1A genes’ expression [48, 60, 61]. Thus, the contribution of the hormonal pathway to the brexpiprazole regulation of liver CYP is also conceivable. Thyroid hormones negatively affect the expression of many CYP enzymes. However, the observed decrease in T4 does not seem to have a high significance for the regulation of CYP genes, because T4 has a much lower affinity for the thyroid receptor than T3 [60]. In the case of CYP2B and CYP2C6 enzymes, there were discrepancies between CYP activity (increase, no change, increase, respectively), enzyme protein (no change of all three), and mRNA level (no change, decrease, no change, respectively), which might be due to some posttranscriptional or posttranslational events occurring during continued treatment with brexpiprazole. It is also not excluded that the reactive metabolites produced by brexpiprazole could contribute to those molecular/biochemical inconsistencies [21].

On the other hand, the mechanism of the inhibition of CYP2E1 expression by the studied antipsychotic requires additional molecular studies, such as determination of HNF-1α, HNF-4α, or WNT/β-catenin pathway [62, 63]. The decline in the CYP2E1 expression and activity found after brexpiprazole treatment may be relevant to the metabolism of fatty acids, including arachidonic acid, as well as several drugs (chlorzoxazone, acetaminophen, isoniazid) and low-molecular-weight xenobiotics, including ethanol and procarcinogens [64].

In conclusion, prolonged administration of brexpiprazole produced a broad-spectrum effect on CYP expression and activity in rat liver, accompanied by alterations in the expression of hepatic transcription factors and in serum hormone concentrations, indicating the involvement of different mechanisms in the enzyme regulation by the investigated antipsychotic. However, our work has some limitations; for example, it is not known whether, in addition to the observed effect on the expression of transcription factors, brexpiprazole can also activate (or inhibit) them. Considering that the investigated CYP1-3 subfamily enzymes are engaged in the endogenous metabolism and biotransformation of most drugs, and that brexpiprazole is administered to patients for a long time, the tested antipsychotic drug may alter not only its own biotransformation, but also the metabolism of vital endogenous substrates and drugs administered concomitantly during long-term treatment (in particular CYP1A, CYP2D, and CYP3A substrates). And this may lead to changes in the pharmacological effect and clinical outcome of applied treatment. Recent pharmacokinetic studies have described a potential drug interaction between brexpiprazole and clozapine in schizophrenic patients [65]. The addition of brexpiprazole to clozapine treatment resulted in a transient decrease in clozapine plasma concentrations and a recurrence of psychotic symptoms after four to six weeks of adjunctive therapy. However, the mechanism of this pharmacokinetic interaction has not yet been elucidated. Because the regulatory mechanisms of the studied genes/proteins differ somewhat between rats and humans, further clinical observations using human experimental models are necessary (e.g., human hepatocyte spheroids, liver organoids, or in vivo metabolic tests) to find out whether prolonged brexpiprazole treatment affects human hepatic transcription factors and CYP enzymes at therapeutic drug concentrations in vivo.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Material 1 (6.9MB, docx)
Supplementary Material 2 (2.9MB, docx)

Abbreviations

AhR

Aryl hydrocarbon receptor

CAR

Constitutive androstane receptor

CORT

Corticosterone

CYP

Cytochrome P450

FXR

Farnesoid X receptor

GH

Growth hormone

HPLC

High-Performance Liquid Chromatography

LXR

Liver X receptor

PPARγ

Peroxisome proliferator-activated receptor γ

PXR

Pregnane X receptor

T3

Triiodothyronine

T4

Thyroxine

Author contributions

P.J.D. and W.A.D. participated in the research design; P.J.D and R.P. conducted the experiments; P.J.D., R.P., and W.A.D. performed data analysis; P.J.D. and W.A.D. wrote the manuscript.

Funding

This work was supported by the grant Preludium 16. 2018/31/N/NZ/0185 from the National Science Centre, Kraków, Poland, and by statutory funds from the Maj Institute of Pharmacology, Polish Academy of Sciences, Kraków, Poland.

Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author upon reasonable request.

Declarations

Competing interests

The authors declare no competing interests.

Footnotes

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

  • 1.Mailman RB, Murthy V. Third generation antipsychotic drugs: partial agonism or receptor functional selectivity? CPD. 2010;16(5):488. 10.2174/138161210790361461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2.Correll CU, Skuban A, Ouyang J, Hobart M, Pfister S, McQuade RD, et al. Efficacy and Safety of brexpiprazole for the treatment of acute schizophrenia: a 6-week Randomized, Double-blind, placebo-controlled Trial. AJP. 2015;172(9):870–80. 10.1176/appi.ajp.2015.14101275. [DOI] [PubMed] [Google Scholar]
  • 3.Orzelska-Górka J, Mikulska J, Wiszniewska A, Biała G. New atypical antipsychotics in the treatment of schizophrenia and depression. IJMS. 2022;23(18):10624. 10.3390/ijms231810624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Ricci V, Sarni A, Martinotti G, Maina G. Comparative analysis of third-generation antipsychotics in first-episode schizophrenia: efficacy, safety, and cognitive impacts. A narrative review. Int Clin Psychopharmacol. 2025;40(4):191–206. 10.1097/YIC.0000000000000559. [DOI] [PubMed] [Google Scholar]
  • 5.Garnock-Jones KP. Brexpiprazole: a review in schizophrenia. CNS Drugs. 2016;30(4):335–42. 10.1007/s40263-016-0325-8. [DOI] [PubMed] [Google Scholar]
  • 6.Watanabe Y, Yamada S, Otsubo T, Kikuchi T. Brexpiprazole for the treatment of schizophrenia in adults: an overview of its clinical efficacy and Safety and a Psychiatrist’s perspective. DDDT 2020, 14: 5559–74. 10.2147/DDDT.S240859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Therrien F, Ward C, Chokka P, Habert J, Ismail Z, McIntyre RS, et al. Adjunctive Brexpiprazole for Patient Life Engagement in Major Depressive Disorder: A Canadian, Phase 4, Open-Label, Interventional Study: Brexpiprazole d’appoint pour l’engagement dans la vie des patients souffrant de trouble dépressif majeur: une étude interventionnelle canadienne ouverte de phase 4. Can J Psychiatry. 2024;69(7):513–23. 10.1177/07067437241233965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Brubaker M, Kapadia S, Maletic V, Chambers JS, Watling CP, Citrome L. Preclinical evidence for the use of brexpiprazole + antidepressant treatment for Major Depressive Disorder and Post-traumatic Stress Disorder: a systematic review. NDT. 2025;21:421–36. 10.2147/NDT.S501207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Giacovelli L, Piccoli E, Landi P, Vismara M, Benatti B, Dell’osso B. Brexpiprazole augmentation in treatment-resistant obsessive-compulsive disorder: a preliminary retrospective observational study. Int Clin Psychopharmacol. 2025;40(6):e14–20. 10.1097/YIC.0000000000000583. [DOI] [PMC free article] [PubMed]
  • 10.Davis LL, Behl S, Lee D, Zeng H, Skubiak T, Weaver S, et al. Brexpiprazole and sertraline combination treatment in posttraumatic Stress Disorder: a Phase 3 Randomized clinical Trial. JAMA Psychiatry. 2025;82(3):218–27. 10.1001/jamapsychiatry.2024.3996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Lee D, Clark ED, Antonsdottir IM, Porsteinsson AP. Brexpiprazole for Agitation Associated with Dementia due to Alzheimer’s Disease. J Am Med Dir Assoc. 2024;25(10):105173. 10.1016/j.jamda.2024.105173. [DOI] [PubMed] [Google Scholar]
  • 12.Imbimbo C, Cotta Ramusino M, Leone S, Mazzacane F, De Franco V, Gatti A, et al. Emerging pharmacological approaches for psychosis and Agitation in Alzheimer’s Disease. CNS Drugs. 2025;39(2):143–60. 10.1007/s40263-024-01133-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Maeda K, Lerdrup L, Sugino H, Akazawa H, Amada N, McQuade RD, et al. Brexpiprazole II: antipsychotic-like and procognitive effects of a novel serotonin-dopamine activity modulator. J Pharmacol Exp Ther. 2014;350(3):605–14. 10.1124/jpet.114.213819. [DOI] [PubMed] [Google Scholar]
  • 14.Maeda K, Sugino H, Akazawa H, Amada N, Shimada J, Futamura T, et al. Brexpiprazole I: in vitro and in vivo characterization of a novel serotonin-dopamine activity modulator. J Pharmacol Exp Ther. 2014;350(3):589–604. 10.1124/jpet.114.213793. [DOI] [PubMed] [Google Scholar]
  • 15.Sasabe H, Koga T, Furukawa M, Matsunaga M, Kaneko Y, Koyama N, et al. Pharmacokinetics and metabolism of brexpiprazole, a novel serotonin-dopamine activity modulator and its main metabolite in rat, monkey and human. Xenobiotica 2021. 2021;51(5):590–604. 10.1080/00498254.2021.1890275. [DOI] [PubMed] [Google Scholar]
  • 16.Sasabe H, Koga T, Furukawa M, Matsunaga M, Sasahara K, Hashizume K, et al. In vitro evaluations for pharmacokinetic drug-drug interactions of a novel serotonin-dopamine activity modulator, brexpiprazole. Xenobiotica 2021. 2021;51(5):522–35. 10.1080/00498254.2021.1897898. [DOI] [PubMed] [Google Scholar]
  • 17.Higashi K, Sasaki T, Aoki K, Sekine D, Maeda K, Shiomi Y, et al. Population pharmacokinetics of brexpiprazole in Japanese healthy subjects and patients with schizophrenia. Drug Metab Pharmacokinet. 2025;62:101057. 10.1016/j.dmpk.2025.101057. [DOI] [PubMed] [Google Scholar]
  • 18.Chen B, Zhang X-D, Wen J, Zhang B, Chen D, Wang S, et al. Effects of 26 recombinant CYP3A4 variants on brexpiprazole metabolism. Chem Res Toxicol. 2020;33(1):172–80. 10.1021/acs.chemrestox.9b00186. [DOI] [PubMed] [Google Scholar]
  • 19.Elmokadem A, Bruno CD, Housand C, Jordie EB, Chow CR, Lesko LJ, et al. Brexpiprazole Pharmacokinetics in CYP2D6 poor metabolizers: using physiologically Based pharmacokinetic modeling to optimize time to effective concentrations. J Sport Hist Clin Pharma. 2022;62(1):66–75. 10.1002/jcph.1946. [DOI] [PubMed] [Google Scholar]
  • 20.Frederiksen T, Areberg J, Raoufinia A, Schmidt E, Stage TB, Brøsen K. Estimating the in vivo function of CYP2D6 Alleles through Population pharmacokinetic modeling of brexpiprazole. Clin Pharma Ther. 2023;113(2):360–69. 10.1002/cpt.2791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.Thakkar D, Kate AS. 1-(Benzo[B]thiophen-4-yl)piperazine ring Induced bioactivation of brexpiprazole in liver microsomes: identification and characterization of reactive conjugates using ultra-high-Performance Liquid Chromatography/Quadrupole time-of-flight Mass spectrometry. Eur J Drug Metab Pharmacokinet. 2020;45(3):393–403. 10.1007/s13318-020-00606-8. [DOI] [PubMed] [Google Scholar]
  • 22.Zanger UM, Schwab M. Cytochrome P450 enzymes in drug metabolism: regulation of gene expression, enzyme activities, and impact of genetic variation. Pharmacol Ther. 2013;138(1):103–41. 10.1016/j.pharmthera.2012.12.007. [DOI] [PubMed] [Google Scholar]
  • 23.Basińska-Ziobroń A, Danek PJ, Daniel WA. The effect of prolonged treatment with antipsychotic drugs on cytochrome P450 − drug metabolizing enzymes. Mechanisms of action and significance for pharmacotherapy. Expert Opin Drug Metab Toxicol. 2025;21(8):921–37. 10.1080/17425255.2025.2517731. [DOI] [PubMed] [Google Scholar]
  • 24.Haduch A, Daniel WA. The engagement of brain cytochrome P450 in the metabolism of endogenous neuroactive substrates: a possible role in mental disorders. Drug Metab Rev. 2018;50(4):415–29. 10.1080/03602532.2018.1554674. [DOI] [PubMed] [Google Scholar]
  • 25.Waxman DJ. P450 gene induction by structurally diverse xenochemicals: central role of nuclear receptors CAR, PXR, and PPAR. Archives Biochem Biophys. 1999;369(1):11–23. 10.1006/abbi.1999.1351. [DOI] [PubMed] [Google Scholar]
  • 26.Handschin C, Meyer UA. Induction of drug metabolism: the role of nuclear receptors. Pharmacol Rev. 2003;55(4):649–73. 10.1124/pr.55.4.2. [DOI] [PubMed] [Google Scholar]
  • 27.Kretschmer XC, Baldwin WS. CAR and PXR: xenosensors of endocrine disrupters? Chemico-Biol Interact. 2005;155(3):111–28. 10.1016/j.cbi.2005.06.003. [DOI] [PubMed] [Google Scholar]
  • 28.Wang H, Negishi M. Transcriptional regulation of cytochrome p450 2B genes by nuclear receptors. CDM. 2003;4(6):515–25. 10.2174/1389200033489262. [DOI] [PubMed] [Google Scholar]
  • 29.Gnerre C, Blättler S, Kaufmann MR, Looser R, Meyer UA. Regulation of CYP3A4 by the bile acid receptor FXR: evidence for functional binding sites in the CYP3A4 gene. Pharmacogenetics. 2004;14(10):635–45. 10.1097/00008571-200410000-00001. [DOI] [PubMed]
  • 30.Gnerre C, Schuster GU, Roth A, Handschin C, Johansson L, Looser R, et al. LXR deficiency and cholesterol feeding affect the expression and phenobarbital-mediated induction of cytochromes P450 in mouse liver. J Lipid Res. 2005;46(8):1633–42. 10.1194/jlr.M400453-JLR200. [DOI] [PubMed] [Google Scholar]
  • 31.Handschin C, Meyer UA. Regulatory network of lipid-sensing nuclear receptors: roles for CAR, PXR, LXR, and FXR. Archives Biochem Biophys. 2005;433(2):387–96. 10.1016/j.abb.2004.08.030. [DOI] [PubMed] [Google Scholar]
  • 32.Hafner M, Rezen T, Rozman D. Regulation of hepatic cytochromes p450 by lipids and cholesterol. CDM. 2011;12(2):173–85. 10.2174/138920011795016890. [DOI] [PubMed] [Google Scholar]
  • 33.Juřica J, Dovrtělová G, Nosková K, Zendulka O. Bile acids, nuclear receptors and cytochrome P450. Physiol Res. 2016;65:S427–40. 10.33549/physiolres.933512. [DOI] [PubMed]
  • 34.Panzitt K, Wagner M. FXR in liver physiology: multiple faces to regulate liver metabolism. Biochim Et Biophys Acta (BBA) - Mol Basis Disease. 2021;1867(7):166133. 10.1016/j.bbadis.2021.166133. [DOI] [PubMed] [Google Scholar]
  • 35.Chen J, Zhao K-N, Chen C. The role of CYP3A4 in the biotransformation of bile acids and therapeutic implication for cholestasis. Ann. Transl. Med. 2014;2:9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36.Chen C, Lin P, Wu Z, Lin Y, Huang M, He L, et al. Farnesoid X receptor regulates CYP1A1 and CYP1B1 and estradiol metabolism in mouse and human cell lines. Chemico-Biol Interact. 2025;412:111471. 10.1016/j.cbi.2025.111471. [DOI] [PubMed] [Google Scholar]
  • 37.Miyamoto S, Miyake N, Jarskog LF, Fleischhacker WW, Lieberman JA. Pharmacological treatment of schizophrenia: a critical review of the pharmacology and clinical effects of current and future therapeutic agents. Mol. Psychiatry. 2012;17(12):1206–27. 10.1038/mp.2012.47. [DOI] [PubMed] [Google Scholar]
  • 38.Schoretsanitis G, Paulzen M, Unterecker S, Schwarz M, Conca A, Zernig G, et al. TDM in psychiatry and neurology: a comprehensive summary of the consensus guidelines for therapeutic drug monitoring in neuropsychopharmacology, update 2017; a tool for clinicians. World J Biol Psychiatry. 2018;19(3):162–74. 10.1080/15622975.2018.1439595. [DOI] [PubMed] [Google Scholar]
  • 39.Ishigooka J, Iwashita S, Higashi K, Liew EL, Tadori Y. Pharmacokinetics and Safety of brexpiprazole following Multiple-dose administration to Japanese patients with schizophrenia. J Sport Hist Clin Pharma. 2018;58(1):74–80. 10.1002/jcph.979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40.Danek PJ, Bromek E, Daniel WA. The influence of long-term treatment with asenapine on liver cytochrome P450 expression and activity in the rat. The Involvement of different mechanisms. Pharmaceuticals (Basel). 2021;14(7):629. 10.3390/ph14070629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 41.Danek PJ, Kuban W, Daniel WA. The effect of chronic iloperidone treatment on cytochrome P450 expression and activity in the rat liver: involvement of neuroendocrine mechanisms. IJMS. 2021;22(16):8447. 10.3390/ijms22168447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 42.Danek PJ, Daniel WA. The novel atypical antipsychotic lurasidone affects cytochrome P450 expression in the liver and peripheral blood lymphocytes. IJMS. 2023;24(23):16796. 10.3390/ijms242316796. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 43.Danek PJ, Daniel WA. The effect of new atypical antipsychotic drugs on the expression of transcription factors regulating cytochrome P450 enzymes in rat liver. Pharmacol Rep. 2024;76(4):895–901. 10.1007/s43440-024-00608-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 44.Nebert DW. Aryl hydrocarbon receptor (AHR): “pioneer member” of the basic-helix/loop/helix per-arnt-sim (bHLH/PAS) family of “sensors” of foreign and endogenous signals. Prog Lipid Res. 2017;67:38–57. 10.1016/j.plipres.2017.06.001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 45.Martignoni M, Groothuis GMM, de Kanter R. Species differences between mouse, rat, dog, monkey and human CYP-mediated drug metabolism, inhibition and induction. Expert Opin Drug Metab Toxicol. 2006;2(6):875–94. 10.1517/17425255.2.6.875. [DOI] [PubMed] [Google Scholar]
  • 46.Lu J, Shang X, Zhong W, Xu Y, Shi R, Wang X. New insights of CYP1A in endogenous metabolism: a focus on single nucleotide polymorphisms and diseases. Acta Pharmaceutica Sin. B. 2020;10(1):91–104. 10.1016/j.apsb.2019.11.016. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 47.Su T. Regulation of the cytochrome P450 2A genes. Toxicol Appl Pharm. 2004;199(3):285–94. 10.1016/j.taap.2003.11.029. [DOI] [PubMed] [Google Scholar]
  • 48.Pascussi J-M, Gerbal-Chaloin S, Duret C, Daujat-Chavanieu M, Vilarem M-J, Maurel P. The tangle of nuclear receptors that controls xenobiotic metabolism and transport: crosstalk and consequences. Annu Rev Pharmacol Toxicol. 2008;48(1):1–32. 10.1146/annurev.pharmtox.47.120505.105349. [DOI] [PubMed] [Google Scholar]
  • 49.Tolson AH, Wang H. Regulation of drug-metabolizing enzymes by xenobiotic receptors: PXR and CAR. Adv Drug Deliv Rev. 2010;62(13):1238–49. 10.1016/j.addr.2010.08.006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 50.Chai SC, Cherian MT, Wang Y-M, Chen T. Small-molecule modulators of PXR and CAR. Biochim Et Biophys Acta (BBA) - Gene Regul Mechanisms. 2016;1859(9):1141–54. 10.1016/j.bbagrm.2016.02.013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 51.Zhang F, Li J, Na S, Wu J, Yang Z, Xie X, et al. The Involvement of PPARs in the selective regulation of brain CYP2D by growth hormone. Neuroscience. 2018;379:115–25. 10.1016/j.neuroscience.2018.03.009. [DOI] [PubMed] [Google Scholar]
  • 52.Ito O, Nakamura Y, Tan L, Ishizuka T, Sasaki Y, Minami N, et al. Expression of cytochrome P-450 4 enzymes in the kidney and liver: regulation by PPAR and species-difference between rat and human. Mol Cell Biochem. 2006;284(1–2):141–48. 10.1007/s11010-005-9038-x. [DOI] [PubMed] [Google Scholar]
  • 53.Paul PR, Mishra MK, Bora S, Kukal S, Singh A, Kukreti S, et al. The impact of P-Glycoprotein on CNS drug efflux and variability in response. J Biochem Mol Tox. 2025;39(3):e70190. 10.1002/jbt.70190. [DOI] [PubMed]
  • 54.Chai AB, Callaghan R, Gelissen IC. Regulation of P-Glycoprotein in the brain. Int J Mol Sci. 2022;23(23):14667. 10.3390/ijms232314667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 55.Wójcikowski J, Daniel WA. The brain dopaminergic system as an important center regulating liver cytochrome P450 in the rat. Expert Opin Drug Metab Toxicol. 2009;5(6):631–45. 10.1517/17425250902973703. [DOI] [PubMed] [Google Scholar]
  • 56.Sadakierska-Chudy A, Haduch A, Rysz M, Gołembiowska K, Daniel WA. The role of brain noradrenergic system in the regulation of liver cytochrome P450 expression. Biochemical Pharmacol. 2013;86(6):800–07. 10.1016/j.bcp.2013.07.017. [DOI] [PubMed] [Google Scholar]
  • 57.Daniel WA, Bromek E, Danek PJ, Haduch A. The mechanisms of interactions of psychotropic drugs with liver and brain cytochrome P450 and their significance for drug effect and drug-drug interactions. Biochemical Pharmacol. 2022;199:115006. 10.1016/j.bcp.2022.115006. [DOI] [PubMed] [Google Scholar]
  • 58.Bromek E, Daniel WA. The regulation of liver cytochrome P450 expression and activity by the brain serotonergic system in different experimental models. Expert Opin Drug Metab Toxicol. 2021;17(4):413–24. 10.1080/17425255.2021.1872543. [DOI] [PubMed] [Google Scholar]
  • 59.Waxman DJ, O’Connor C. Growth hormone regulation of sex-dependent liver gene expression. Mol Endocrinol. 2006;20(11):2613–29. 10.1210/me.2006-0007. [DOI] [PubMed] [Google Scholar]
  • 60.Monostory K, Pascussi J-M, Kóbori L, Dvorak Z. Hormonal regulation of CYP1A expression. Drug Metab Rev. 2009;41(4):547–72. 10.1080/03602530903112284. [DOI] [PubMed] [Google Scholar]
  • 61.Monostory K, Dvorak Z. Steroid regulation of drug-metabolizing cytochromes P450. CDM. 2011;12(2):154–72. 10.2174/138920011795016854. [DOI] [PubMed] [Google Scholar]
  • 62.Kocarek TA, Zangar RC, Novak RF. Post-transcriptional regulation of rat CYP2E1 expression: role of CYP2E1 mRNA untranslated regions in control of translational efficiency and message stability. Archives Biochem Biophys. 2000;376(1):180–90. 10.1006/abbi.2000.1704. [DOI] [PubMed] [Google Scholar]
  • 63.Liu Y, Xu Q, Liu Y, Cao S, Luo J, Zheng Z, et al. Hepatocyte-targeted Lipid nanoparticle delivery of HERC2 plasmid controls drug-Induced hepatotoxicity by limiting β-catenin-regulated CYP2E1 expression. Adv Sci. 2024;11(46):e2401633. 10.1002/advs.202401633. [DOI] [PMC free article] [PubMed]
  • 64.Chen J, Jiang S, Wang J, Renukuntla J, Sirimulla S, Chen J. A comprehensive review of cytochrome P450 2E1 for xenobiotic metabolism. Drug Metab Rev. 2019;51(2):178–95. 10.1080/03602532.2019.1632889. [DOI] [PubMed] [Google Scholar]
  • 65.Cereda G, Lazzaretti M, Colli C, Fontana E, Enrico P, Pigoni A, et al. Pharmacokinetic drug-drug interaction between clozapine and brexpiprazole: a case series. Schizophr Res. 2023;261:15–17. 10.1016/j.schres.2023.08.028. [DOI] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Supplementary Material 1 (6.9MB, docx)
Supplementary Material 2 (2.9MB, docx)

Data Availability Statement

The datasets generated during and/or analysed during the current study are available from the corresponding author upon reasonable request.

Articles from Pharmacological Reports are provided here courtesy of Springer

RESOURCES