BACKGROUND:Efsubaglutide Alfa is a novel long-acting glucagon-like peptide-1 receptor agonist developed to promote weight loss and improve metabolic outcomes. This Phase 2a trial evaluated its efficacy, tolerability, safety, and pharmacokinetics (PK) in overweight and obese individuals unresponsive to lifestyle interventions. METHODS:In this multicenter, randomized, double-blind, placebo-controlled, multiple-ascending dose study, 50 participants were randomized (8:2) across five dose cohorts (5, 7.5, 10, 15, and 20 mg) or placebo. Participants received once-weekly subcutaneous injections of Efsubaglutide Alfa or placebo, with individualized dose escalation every 2 weeks, followed by 4 weeks at target doses. Primary endpoints were percentage change in body weight from baseline and the proportion achieving ≥5% weight loss. Secondary endpoints included changes in body composition and a metabolic composite index (BMI, waist circumference, blood pressure, lipid profiles). Tolerability, safety, immunogenicity, and PK were assessed. FINDINGS:Between 25 April 2024 and 18 November 2024, 50 individuals were randomly assigned. Mean baseline characteristics included age 36.3 years, bodyweight 92.9 kg, and BMI 33.0 kg/m2. Efsubaglutide Alfa produced a mean weight reduction of 7.16% (95% CI: -8.08 to -6.24) versus 0.86% with placebo. Overall, 82.5% of Efsubaglutide-treated participants achieved ≥5% weight loss (vs. 0% placebo). Fat mass decreased by 4.47 kg, and lean mass also declined by 2.00 kg from baseline; however, the lean-to-fat mass ratio improved by 19.73 percentage points. BMI, waist circumference, and systolic blood pressure significantly decreased. Gastrointestinal adverse events were the most common, mostly mild to moderate, occurring primarily during dose escalation. No treatment-related serious adverse events occurred. Efsubaglutide Alfa showed dose-proportional PK. INTERPRETATION:Efsubaglutide Alfa demonstrated significant weight-loss efficacy, metabolic improvements, and a preferable tolerability and safety profile, supporting further clinical development for obesity and related metabolic disorders.

Vebreltinib is a novel, highly selective type I c-Met inhibitor developed for treating non-small cell lung cancer (NSCLC) and glioma. As in vitro studies indicate that vebreltinib is primarily metabolized via cytochrome P450 (CYP) 3A4, this study aimed to characterize the drug-drug interaction (DDI) potential of vebreltinib with strong CYP3A4 modulators. Following a preliminary single ascending dose (SAD) study to establish baseline safety and pharmacokinetics (PK) in healthy participants, a DDI study was designed to evaluate the impact of the strong CYP3A4 inducer rifampin and the strong CYP3A4 inhibitor itraconazole on vebreltinib exposure. In the CYP3A4 induction part of the DDI study, participants (n = 14) received 200 mg vebreltinib on Days 1 and 15, with rifampin (600 mg once daily) administered on Days 8-21. In the inhibition part, participants (n = 14) received 200 mg vebreltinib on Days 1 and 11. Itraconazole was administered as a loading dose (200 mg BID on Day 8) followed by maintenance dosing (200 mg QD from Days 9 to 19). PK blood samples were collected at predetermined time points. Vebreltinib plasma concentrations were assessed using a validated liquid chromatography-tandem mass spectrometry (LC-MS) method. Co-administration with rifampin reduced the AUC0-t and AUC0-∞of vebreltinib by ~65% and ~66%, respectively, whereas the Cmax did not change significantly. In contrast, coadministration with itraconazole led to an increase in the AUC and Cmax of vebreltinib by about 56% and 46%, respectively. Vebreltinib, both alone and in combination with rifampin or itraconazole, was well tolerated in healthy participants.

Background:The global obesity crisis requires precision biomarkers to overcome treatment resistance. We investigated circulating multi-omics signatures for predicting and monitoring glucagon-like peptide-1 receptor agonist (GLP-1RA) (GZR18) response, addressing gaps in personalized obesity therapy. Methods:We conducted longitudinal multi-omics profiling (proteomics, metabolomics, and lipidomics) of 221 plasma samples from 25 participants (n=25) treated with GZR18 at nine time points over 30 weeks (four samples could not be tested due to hemolysis). High-resolution mass spectrometry quantified molecular features alongside clinical body mass index (BMI) trajectories. Theil-Sen regression modeled baseline predictors (BMI slope Z), while linear regression analysis identified longitudinal biomarkers (%BMI change). Significant candidates (P<0.05) underwent gene set enrichment analysis (GSEA; Kyoto Encyclopedia of Genes and Genomes [KEGG] pathways) and STRING network integration, with dual-response biomarkers validated through correlation and trajectory analyses. Results:GZR18-treated obese patients exhibited a dose-dependent reduction in BMI, with 48 mg biweekly producing the steepest declines (Z<-0.4 vs. placebo: Z=-0.22, P<0.001). We identified glycogenin 1 (GYG1) as a dual-function biomarker (coefficient=2.0 for prediction, P<0.05 for monitoring) and as a central network hub. Lipidomic (phosphatidylinositol 18:2-18:2) and metabolomic (oxoglutaric acid) markers predicted baseline response, while proteomic (insulin like growth factor binding protein 2 [IGFBP2]) and lipidomic (phosphatidylcholine 18:0-20:3) profiles tracked longitudinal efficacy (P<0.05). Adipocytokine signaling governed the initial response (normalized enrichment score >1.8), while starch/sucrose metabolism modulated ongoing efficacy through integrated molecular networks. Conclusion:This study establishes GYG1 as a clinically actionable, dual-function biomarker for GLP-1RA therapy in obesity, linking baseline prediction and real-time monitoring of treatment response through integrated multi-omics profiling. Our findings highlight convergent metabolic pathways driving therapeutic efficacy and provide a precision-medicine framework for optimizing obesity pharmacotherapy through blood-based molecular signatures.

GZR18 is a glucagon-like peptide-1 receptor agonist under development for overweight/obesity. In this randomized, placebo-controlled phase 1b/2a trial in Chinese adults with overweight or obesity, different dose titration regimens and dosing frequencies (once-weekly [QW] vs. bi-weekly [Q2W]) are evaluated to assess safety and weight-loss efficacy over 26 weeks (Part A) or 35 weeks (Part B). Sixty participants are enrolled, of whom 46 complete the trial. The least-squares mean change in body weight is-9.36% for GZR18 vs. 6.68% for placebo in Part A and-17.8% (QW) and-12.8% (Q2W) for GZR18 vs. 0.7% for placebo in Part B; GZR18 also improves other weight-related and metabolic parameters. GZR18 is generally safe and well tolerated, with mostly mild-to-moderate gastrointestinal adverse events and no investigational-product-related serious adverse events reported. These findings warrant larger and longer trials of GZR18 for body weight management. The trial is registered at ClinicalTrials.gov (NCT06256536).

BACKGROUND:Vebreltinib is a novel, highly selective inhibitor of the hepatocyte growth factor receptor (HGFR, also known as c-MET or MET) tyrosine kinase under development for non-small cell lung cancer. This study aimed to characterize the absorption, metabolism, excretion and mass balance of [14C]-vebreltinib in humans. METHODS:In this open-label study, six healthy Chinese male subjects received a single 200 mg (100 μCi) oral dose of [14C]-vebreltinib. Serial blood was collected for up to 216-h post-dose. Serial urine and faeces were collected for up to 240-h post-dose. Total radioactivity was measured by liquid scintillation counting, and metabolite profiling was conducted using HPLC-radiochromatography coupled with high-resolution mass spectrometry. RESULTS:Vebreltinib was slowly absorbed, with a mean plasma elimination half-life of 20.1 ± 5.3 h. The half-life of total radioactivity was longer, consistent with the formation and persistence of metabolites. The mean total recovery of radioactivity was 93.0% ± 2.1%, with the majority recovered in faeces (79.5% ± 5.5% of the dose) and 13.5% ± 4.3% recovered in urine. Unchanged vebreltinib was the most abundant drug-related component in excreta, with the N-demethylated metabolite M2 as the major circulating metabolite. CONCLUSION:Following oral administration, the predominant radioactive component recovered in faeces was unchanged parent drug. The quantitative metabolic profile confirms significant systemic exposure to the M2 metabolite. These data provide critical insights into the human disposition of vebreltinib, including a MIST assessment, and support its continued clinical development.

BACKGROUND AND PURPOSE:Tyrosine kinase 2 (Tyk2) inhibitors hold therapeutic promise for patients with inflammatory conditions driven by interleukin (IL)-12/23 signalling. ARTS-011 [6-(cyclopropanecarboxamido)-4-((2-methoxy-3-(3-methyl-1H-1,2,4-triazol-1-yl)phenyl)amino)-N-(methyl-d3)pyridazine-3-carboxamide] is a novel chemical agent that acts as a small-molecule allosteric inhibitor of Tyk2, selectively targeting the JH2 domain of the Tyk2 protein. A first-in-human, single and multiple ascending dose (SAD and MAD) clinical study was conducted to evaluate the tolerability, safety, pharmacokinetic profile and food effects of the Tyk2 inhibitor ARTS-011. EXPERIMENTAL APPROACH:This study is registered at ClinicalTrials.gov under the identifier NCT06260527. The study consisted of three parts: - Part A was a SAD cohort and Part B was a MAD cohort both conducted in Chinese healthy subjects, using a randomized, double-blind, placebo-controlled design. Part C was a randomized, open-label, two-period, crossover study designed to investigate the food effects on ARTS-011. KEY RESULTS:Single oral doses ranging from 3 to 60 mg and multiple oral doses ranging from 10 to 40 mg of ARTS-011 demonstrated favourable safety and tolerability profiles in healthy subjects. Furthermore, ARTS-011 was well-tolerated under both fasting and fed conditions following a single 20 mg dose. The 90% confidence interval of the geometric mean ratio of area under the concentration-time curve (AUC) fell entirely within the bioequivalence criteria (80.00%-125.00%), indicating no food effect on AUC. CONCLUSION AND IMPLICATIONS:ARTS-011 is a promising oral Tyk2 inhibitor with an acceptable safety profile and potential for once-daily dosing. These results support further clinical development of ARTS-011 in patients with inflammatory and autoimmune diseases.

ABSTRACT Background Ulcerative colitis (UC) and Helicobacter pylori (H. pylori) infection show an intriguing inverse epidemiological association, but the underlying molecular mechanisms remain unclear, with emerging evidence suggesting H. pylori may modulate colonic inflammation via systemic immune regulation. Methods We used bioinformatics approaches, including gene set enrichment analysis (GSEA), differential expression analysis, functional enrichment (GO/KEGG), protein‐protein interaction (PPI) networks, upstream regulatory molecule prediction, and immune infiltration characterization, to analyze gene expression datasets from UC and H. pylori‐infected samples, aiming to identify the interconnections and regulatory networks between these two conditions. Results GSEA identified 53 shared pathways, primarily innate immune response pathways (e.g., TLR/NLR signaling, NF‐κB/IL‐17 cascades). We found 243 co‐differentially expressed genes enriched in leukocyte chemotaxis, cytokine activity, and extracellular matrix organization. Six hub genes (CXCL8, IL1B, MMP9, CXCL1, IFNG, CXCL9) were validated as robust diagnostic markers (AUC > 0.815 for both conditions). Immune landscape analysis revealed pan‐infiltration of immune cells in H. pylori‐infected tissues and shared dysregulated immune cells in UC tissues, with hub genes positively correlated with immune cell infiltration in both. We also identified regulatory miRNAs (e.g., miR‐204‐5p) and transcription factors (FOXC1, YY1) modulating these hub genes. Conclusion This study uncovers shared immune‐mediated pathways and hub genes linking H. pylori infection to UC, establishing a molecular framework. These hub genes and regulatory networks may serve as diagnostic biomarkers and therapeutic targets, highlighting the need to investigate H. pylori‐driven immune modulation in UC pathogenesis.

GZR18 is a long-acting glucagon-like peptide-1 receptor agonist that is under development for the treatment of type 2 diabetes mellitus (T2DM). In this randomized, double-blind, placebo-controlled phase 1b/2a trial in Chinese adults with T2DM, the safety and efficacy of once-weekly GZR18 are evaluated using different dose-titration regimens in two separate parts: Part A (0.5-4 mg over 26 weeks) and Part B (1.5-13 mg over 23 weeks). Overall, 72 participants are enrolled, of whom 62 complete the trial. GZR18 significantly reduces glycated hemoglobin (HbA1c) in both parts (Part A: -1.32% for GZR18 versus 0.86% for placebo; Part B: -1.81% for GZR18 versus 0.12% for placebo). Improvements in body weight and other metabolic parameters are also observed. GZR18 is safe and well tolerated, with mostly mild-to-moderate gastrointestinal adverse events; no severe hypoglycemia or treatment-related serious adverse events occurred. These findings support further development of GZR18 in larger, longer-term trials. The trial is registered at ClinicalTrials.gov (NCT06256523).

Bofanglutide is a novel biweekly (once every two weeks; Q2W) glucagon-like peptide-1 receptor agonist. We evaluated the efficacy and safety of bofanglutide in Chinese adults with overweight or obesity in a randomized, double-blind, placebo-controlled phase 2b trial (ClinicalTrials.gov, NCT06256562). Adults with overweight (body mass index [BMI] ≥24, <28 kg/m2) and at least one weight-related comorbidity, or obesity (BMI ≥ 28 kg/m2), were randomly assigned to five dose groups: 12 mg Q2W, 18 mg Q2W, 24 mg Q2W, 48 mg Q2W, and 24 mg once weekly (QW), with randomization to bofanglutide or placebo within each dose group. The primary endpoint was the percentage change in body weight from baseline to week 30. Between June 8, 2023, and June 5, 2024, 340 participants (185 [54.4%] male; mean age, 33.1 years; mean body weight, 95.6 kg; mean BMI, 33.2 kg/m²) were randomized into the following groups: bofanglutide 12 mg Q2W (n = 52), 18 mg Q2W (n = 53), 24 mg Q2W (n = 52), 48 mg Q2W (n = 64), 24 mg QW (n = 53), or placebo (n = 66). Overall, 286 participants (84.1%) completed the trial. The mean percentage change in body weight from baseline to week 30 ranged from −9.75% to −16.69% with bofanglutide, compared with −1.15% with placebo (all p < 0.001 versus placebo). Adverse events occurred in 98.9% (271/274) of the bofanglutide group versus 86.4% (57/66) of the placebo group and were mostly grade 1–2 gastrointestinal events (83.9% [230/274] with bofanglutide and 33.3% [22/66] with placebo). Bofanglutide is generally well tolerated and has a robust ability to reduce body weight.

This study aimed to investigate cervical vessel hemodynamics during 7 days of −6° head-down tilt (HDT) bed rest (BR) and 5 days of recovery in 40 healthy males, focusing on bilateral analysis and dynamic temporal monitoring. Diameter, flow velocity, and blood flow volume (BFV) of bilateral common (CCAs), internal (ICAs), external carotid arteries and vertebral arteries (VAs) were measured at eight time points using ultrasound. Cervical vessels exhibited heterogeneous and lateralized responses. Left VA exhibited the most pronounced BFV and velocity fluctuations, with significant reductions during HDT and recovery to baseline post-HDT, while right VA remained stable. CCAs and ICAs had dynamic diameter and velocity changes, but stable BFVs. Left VA BFV reduction was significantly associated with right VA dominance, elevated heart rate, and increased mean arterial pressure at HDT 3 d. Despite stable total BFV, left VA BFV declined during BR, warranting further monitoring.

BackgroundThe bioequivalence of Concor® (Merck Healthcare KGaA, Darmstadt, Germany), a bisoprolol-containing tablet, manufactured in China and Concor® tablets manufactured in Germany has not been previously reported.MethodsThis single-center, open-label, randomized, two-period, two-sequence, crossover trial (28 February 2023–19 May 2023) compared the pharmacokinetics and safety of bisoprolol 5-mg tablets manufactured in China (test product) with those of bisoprolol 5-mg tablets manufactured in Germany (reference product) in healthy Chinese adults under fasted and fed conditions. Primary endpoints were Cmax, AUC0–tlast, and AUC0–∞.ResultThe mean (coefficient of variation percentage) Cmax in the fasted group was 21.2 (15.0) ng/mL (test product) and 22.1 (17.0) ng/mL (reference product). Under fed conditions, the respective Cmax values were 22.7 (18.8) ng/mL and 22.8 (15.2) ng/mL. The mean and coefficient of variation percentage for AUC were also similar between the two products. The geometric least squares mean ratio (90% confidence interval) for the test/reference product was 0.9565 (0.9006–1.0158) ng/mL for Cmax, 0.9761 (0.9370–1.0168) h·ng/mL for AUC0–tlast, and 0.9807 (0.9429–1.0200) h·ng/mL for AUC0–∞ in fasted conditions and 0.9966 (0.9289–1.0691) ng/mL for Cmax, 0.9672 (0.9220–1.0145) h·ng/mL for AUC0–tlast, and 0.9693 (0.9253–1.0155) h·ng/mL for AUC0–∞ in fed conditions, which met the pre-defined criteria for bioequivalence. No serious treatment-emergent adverse events or deaths were observed.ConclusionThis study compared the bioequivalence of bisoprolol 5-mg tablets manufactured in China to that of the tablets manufactured in Germany among healthy Chinese adults.Systematic Review Registrationidentifier CTR20230391

HT-101, a liver-targeted N-acetylgalactosamine-conjugated ribonucleic acid interference therapeutic, exhibits promising potential for the treatment of chronic hepatitis B virus infection. This randomized, double-blind, placebo-controlled, and single-ascending-dose Phase Ia study included 50 healthy volunteers. Regarding methods, 2 subjects received a single subcutaneous dose of HT-101 at 25 mg, while 48 volunteers were randomized (6:2 active:placebo) in the remaining 6 cohorts to receive a single subcutaneous dose of HT-101 (50-800 mg) or placebo. Afterward, serial blood samples were obtained for pharmacokinetic determination across a 48-hour postdose period. Safety assessments included clinical laboratory measures, vital signs, and 12-lead electrocardiogram before and after dosing. As a result, plasma pharmacokinetics characterized by functional antisense strand revealed a median time to peak plasma concentration of 2.5-6.0 hours, and a short median plasma half-life of 2.50-6.14 hours. It is underlined that peak and total plasma exposure to HT-101 increased in a slightly greater-than-dose-proportional manner following 25-800 mg administered subcutaneously. Moreover, a single dose of HT-101 at 25-800 mg was safe and well tolerated in healthy Chinese volunteers. These data can support further clinical development of HT-101 for hepatitis B virus infection treatment.

Bruton’s tyrosine kinase (BTK) inhibitors play a critical role in the treatment of mantle cell lymphoma (MCL). pirtobrutinib, a new, highly selective, non-covalent BTK inhibitor, was approved by the FDA for the treatment of MCL, chronic lymphocytic leukemia (CLL), and small lymphocytic lymphoma (SLL). In this study, we established a robust and reliable method for the quantitation of pirtobrutinib in rat plasma using ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Acetonitrile and 0.1

BACKGROUND:Our objective was to evaluate the safety and immunogenicity of the COVID-19 mRNA vaccine (CS-2034) in Chinese adults, and to determine the optimal dosage with a favorable safety and immunogenicity profile. METHODS:This study was a phase II clinical trial conducted across multiple centers, employing a randomized, double-blinded, dose-exploration, placebo-controlled design which was registered at ClinicalTrials.gov under the identifier NCT05373472 (registration date: May 13, 2022). RESULTS:A total of 150 participants were randomized into low-dose vaccine, high-dose vaccine, and placebo groups. The vaccine CS-2034 was well tolerated, with adverse reactions largely mild to moderate and comparable across dose groups. Both dose levels induced strong neutralizing antibody responses against the ancestral SARS-CoV-2 strain, with high seroconversion and evidence of antibody persistence over time. In contrast, neutralizing activity against the Omicron BA.1 variant was limited and declined further after the peak response. CONCLUSIONS:The 0.3 mL dose (low-dose vaccine) was selected for further development based on comparable immunogenicity and lower reactogenicity. CLINICAL TRIAL REGISTRATION:The study was registered at ClinicalTrials.gov under the identifier NCT05373472 (registration date: May 13, 2022).

The number of drug-drug interaction (DDI) clinical trials in China has increased rapidly in recent years. The aim of this study was to summarize and analyze DDI clinical trials in China over the past 10 years. We conducted a cross-sectional study of DDI clinical trials registered in the Chinese Center for Drug Evaluation (CDE) from September 6, 2013 to December 31, 2022. All related registration information disclosed on the CDE website were summarized and analyzed. Although the number of DDI clinical trials conducted before 2017 was relatively low, it increased markedly after 2017. The average duration of DDI clinical trials was 85.83 ± 100.99 days from 2013 to 2019 and 107.16 ± 98.57 days from 2020 to 2022. The duration of rifampicin use was 5–19 days, and the investigational drug was administered after 5–14 days of rifampicin use. Itraconazole was administered for 4–17 days, and the investigational drug was administered after 3–10 days of itraconazole use. Clinical trials of drug-drug interactions have recently increased due to the development of new drugs and the updated policies regulating drug registration and marketing. Although the designs of clinical trials comply with the new guidelines, the duration of the administration of interacting drugs still varies widely. Optimizing protocol designs can shorten the implementation period of clinical trials and reduce the costs of drug marketing.

IntroductionGlutathione peroxidase 4 (GPX4) is a key enzyme in ferroptosis. Gaining insight into GPX4’s mechanisms and biological roles could offer valuable therapeutic insights for cancer treatment.MethodsBy integrating multi-omics data from The Cancer Genome Atlas (TCGA), the Genotype-Tissue Expression Project (GTEx), cBioPortal, the Human Protein Atlas (HPA), UALCAN, Xiaotao platform and et al., we applied systematic bioinformatics approaches to evaluate the expression, prognostic significance, mutation profiles, DNA methylation and tumor immune microenvironment (TIME) infiltration of GPX4 across diverse cancer types. Furthermore, the role of GPX4 in cell proliferation was experimentally validated.ResultsGPX4 was upregulated in several cancer types. Its potential as a diagnostic biomarker was confirmed by its high reliability in differentiating cancerous from normal tissues, with AUC values surpassing 0.8 in multiple cancers. Functional studies verified its oncogenic function in colorectal and gastric cancer cell lines. In terms of prognosis, GPX4 expression levels were closely associated with overall survival across various cancers. Furthermore, we detected a correlation between the mutation burden of GPX4 across different types of cancer and patient survival outcomes. Additionally, immune infiltration analysis showed significant correlations between GPX4 expression and immune cell presence, particularly macrophages and M2 type macrophages. GPX4 expression also correlated highly with immune modulator pathways and checkpoints. ConclusionCollectively, these pan-cancer analyses underscore the potential of GPX4 as a therapeutic target and biomarker in multiple cancers. Further indepth studies on GPX4’s regulatory mechanisms and clinicopathological significance are warranted to develop novel therapies for the prevention and treatment of human tumors.

AbstractSKLB1028 is a novel multi‐target protein kinase inhibitor under investigation for the treatment of FLT3‐ITD mutated acute myeloid leukemia. Based on the preclinical characterization of SKLB1028 metabolism, three drug–drug interaction clinical studies were performed to investigate the effects of itraconazole, rifampin (CYP3A4 inhibitor and inducer, respectively), and gemfibrozil (CYP2C8 inhibitor) on the metabolism of SKLB1028. Fourteen healthy Chinese male subjects were enrolled in each study. In Study 1, subjects were administered a single dose of SKLB1028 (100 mg on days 1 and 11) and multiple doses of itraconazole (200 mg twice daily on day 8 and 200 mg once daily from days 9 to 18). Itraconazole was given with a loading dose on Day 8 and the total administration of itraconazole was 11 days. In Study 2, subjects were administered a single dose of SKLB1028 (100 mg on days 1 and 12) and multiple doses of gemfibrozil (600 mg twice daily from days 8 to 19). In Study 3, subjects were administered a single dose of SKLB1028 (150 mg on days 1 and 15) and multiple doses of rifampin (600 mg once daily from day 8 to 22). Itraconazole increased the AUC and Cmax of SKLB1028 by approximately 28% and 41%, respectively. Compared to the single drug, co‐administration with gemfibrozil increased the AUC of SKLB1028 by ~26% and the Cmax by ~21%. Co‐administration with rifampin reduced the AUC of SKLB1028 by ~30%, while the Cmax did not change significantly. All treatments were well tolerated in all three studies.

AbstractA drug–drug interaction (DDI) trial of cytochrome P450 3A (CYP3A) is a necessary part of early‐phase trials of drugs mainly metabolized by this enzyme, but CYP3A DDI clinical trials do not have a standard design, especially for Chinese people. We aimed to offer specific recommendations for CYP3A DDI clinical trial design. This was an open, three‐cycle, self‐controlled study. Healthy subjects were given different administration strategies of CYP3A4 perpetrators. In each cycle, blood samples were collected before and within 24 h after the administration of midazolam, the CYP3A indicator substrate. The plasma concentrations of midazolam and 1‐hydroxymidazolam was obtained using liquid chromatography tandem mass spectrometry assay. For CYP3A inhibition, itraconazole exposure with a loading dose could increase the exposure of midazolam by 3.21‐fold based on maximum plasma concentration (Cmax), 8.37‐fold based on area under the curve Pharmacology Research & Perspectives for review only from zero to the time point (AUC0–t), and 11.22‐fold based on area under the curve from zero to infinity (AUC0–∞). The data were similar for itraconazole pretreatment without a loading dose. For CYP3A induction, the exposure of rifampin for 7 days decreased the plasma concentration of midazolam ~0.27‐fold based on Cmax, ~0.18‐fold based on AUC0–t, and ~0.18‐fold based on AUC0–∞. Midazolam exposure did not significantly change when the pretreatment of rifampin increased to 14 days. This study showed that itraconazole pretreatment for 3 days without a loading dose was enough for CYP3A inhibition, and pretreatment with rifampin for 7 days could induce near‐maximal CYP3A levels.