A retrospective analysis of patient-specific factors on voriconazole clearance
© Dote et al. 2016
Received: 18 December 2015
Accepted: 28 March 2016
Published: 19 April 2016
Voriconazole concentrations display a large variability, which cannot completely be explained by known factors. We investigated the relationships of voriconazole concentration with patient-specific variables and concomitant medication to identify clinical factors affecting voriconazole clearance.
A retrospective chart review of voriconazole trough concentration, laboratory data, and concomitant medication in patients was performed. The concentration/dose ratio (C/D-ratio) was assessed as a surrogate marker of total clearance by dividing voriconazole concentration by daily dose per kg of body weight.
A total of 77 samples from 63 patients were obtained. In multiple linear regression analysis, increased C-reactive protein (CRP) level (p < 0.05) and decreased albumin (Alb) level (p < 0.05) were associated with significantly increased C/D-ratio of voriconazole, and coadministration with a glucocorticoid was associated with significantly (p < 0.05) decreased C/D-ratio of voriconazole (adjusted r 2 = 0.31). Regarding CRP and Alb, receiver operating characteristic curve analysis indicated that increased CRP level and decreased Alb level were significant predictors of toxic trough concentration of voriconazole. For CRP, area under the curve (AUC) and cutoff value were 0.71 (95 % confidence interval (CI), 0.57–0.86, p < 0.01) and 4.7 mg/dl, respectively. For Alb, AUC and cutoff value were 0.68 (95 % CI, 0.53–0.82, p < 0.05) and 2.7 g/dl, respectively. A significant difference was seen in voriconazole trough concentration between patients with hepatotoxicity and those without (5.69 μg/ml vs 3.0 μg/ml, p < 0.001).
Coadministration of glucocorticoid and inflammation, reflected by elevated CRP level and hypoalbuminemia, are associated with voriconazole clearance. We propose that early measurement of voriconazole concentration before the plateau phase will lead to avoidance of a toxic voriconazole level in patients with elevated CRP level and hypoalbuminemia, although further studies are needed to confirm our findings.
KeywordsVoriconazole Pharmacokinetics Therapeutic drug monitoring Drug interaction Inflammation
The triazole antifungal voriconazole is widely used in the treatment of invasive fungal infections, such as invasive aspergillosis, candidemia, and pulmonary cryptococcosis, except for zygomycosis [1–4]. Voriconazole exhibits a nonlinear pharmacokinetics profile and is metabolized primarily by the cytochrome P450 family enzyme CYP2C19 and, to lesser extent, CYP3A4 and CYP2C9 [1–4]. Allelic polymorphisms of CYP2C19 have been shown to be the most important determinants of the clearance efficiency of vorizonazole [3, 5]. In Japanese, the rate of poor metabolizers of CYP2C19 has been shown to be 15–20 %, a higher figure than that of other races . This raises the necessity of blood concentration measurement for Japanese to monitor efficacy and toxicity in the clinical setting. There have been some reports on the relationships between efficacy and toxicity of voriconazole and its plasma concentrations. Target trough concentration of therapeutic response is > 1–2 μg/ml [1, 3, 4, 7–9], and prophylaxis is > 1.5 μg/ml . To avoid side effects, such as visual disturbance, neurotoxicity, and hepatotoxicity, the target trough concentration is < 4–6 μg/ml [1, 3, 4, 7–10]. Invasive fungal infections commonly appear among immunocompromised patients receiving concomitant medication for primary disease and/or bacterial infection. Many of them have critically ill conditions and polypharmacy, which possibly alter voriconazole concentration and clearance. Accordingly, to identify clinical factors affecting voriconazole clearance, we investigated the relationships of voriconazole concentration with patient-specific variables and concomitant medication.
Patient enrollment and data collection
Patients who received voriconazole orally and had at least one voriconazole trough concentration measurement during therapy at Kyoto-Katsura Hospital, Japan, between June 2010 and October 2015 were eligible for inclusion. Patient-specific variables were collected retrospectively including sex, age, body weight, body mass index (BMI), laboratory data, purpose of voriconazole administration, and diagnosis and treatment department. Laboratory data included serum albumin (Alb), creatinine clearance (Ccr), aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (T-Bil), alkaline phosphatase (ALP), γ-glutamyl transpeptitase (γGTP), and C-reactive protein (CRP). Ccr was estimated by using the Cockcroft-Gault Formula . Laboratory data at voriconazole trough concentration measurement were used for analysis. Daily dose, voriconazole trough concentration, the number of days between blood sample collection and initial dose were collected. Patients were excluded if: (a) they received voriconazole intravenously even once to avoid unknown covariates between orally and intravenously; (b) they received concomitant rifabutin, rifampicin, carbamazepine, phenytoin, or antiretroviral drugs; (c) they required dialysis since the uremic substances, which should accumulate in the blood of those patients, may alter the pharmacokinetics of voriconazole; or (d) the number of days between blood sample collection and initial voriconazole administration with loading dose is 2 or less, or that without loading dose is 4 or less, because the concentration did not reach a steady state [3, 4]. Affect of concomitant glucocorticoid (over 5 mg equivalent prednisolone as a daily dose) and macrolide was analyzed in all patients, because those medications can interact with voriconazole pharmacokinetically [9, 12, 13]. Cases in which the voriconazole dose was changed, or the above-mentioned concomitant medications were started or discontinued, were regarded as separate cases. Visual disturbance such as photophobia, blurred vision, neurotoxicity such as drowsiness, insomnia, hallucinations, disturbance of consciousness, and hepatotoxicity were evaluated as adverse events. Hepatotoxicity was defined as ≥ grade 2 in results of liver function tests (AST, ALT, T-Bil, ALP, and γGTP) within 14 days after initiation of voriconazole administration or changing voriconazole dosage, according to Common Terminology Criteria for Adverse Events (CTCAE), version 4.0, since some of ALP and γGTP values in the patients were over normal range (about grade 1) before the voriconazole therapy. We used voriconazole concentration at the time of occurrence of toxicity in the patients with adverse event, whereas the maximum concentration in the patients without adverse event for the analysis of relationship between adverse events and voriconazole concentration. All voriconazole concentration data were collected from a central referral laboratory (SRL Inc., Tokyo, Japan). A validated high-performance liquid chromatography (HPLC) assay was used to measure voriconazole concentrations .
The study was performed in accordance with the Declaration of Helsinki and its amendments, and the protocol was approved by the Ethics Committee of Kyoto-Katsura Hospital.
Analysis of voriconazole pharmacokinetics
The concentration/dose ratio (C/D-ratio) was used as a surrogate marker of total clearance by dividing voriconazole concentration by daily dose per kg of body weight.
Univariate linear regression analysis was used to estimate relationships between C/D-ratio of voriconazole and patient-specific continuous variables, and the unpaired t test was used to compare voriconazole concentration and C/D-ratio of voriconazole between two independent groups. Multiple linear regression analysis was applied to identify factors that contribute to the variability in C/D-ratio of voriconazole; a step-down selection model was obtained using stepwise regression with a p value of 0.25 as the inclusion criterion. The diagnostic properties and cutoff values of independent variables by multiple linear regression analysis were cross-sectionally evaluated by a receiver operating characteristic (ROC) curve. Outcome for diagnostic properties was voriconazole trough concentration exceeding 4 μg/ml, defined as a toxic level according to the results of meta-analysis by Hamada et al. . JMP®9 software was used for all analyses and a p value less than 0.05 was regarded as statistically significant.
Body Weight (kg)
Body Mass Index (kg/m2)
Creatinine clearance (ml/min)a
C reactive protein (mg/dl)a
Daily dose (mg/day)a
Trough concentration (μg/ml)a
Trough concentration collection date (after initial dose)a
with loading dose
without loading dose
Purpose of voriconazole administration prophylaxis/treatment
Diagnosis and treatment department
Factors affecting voriconazole concentration and clearance
Factors associated with a significant change in vorizconazole clearance identified from multiple linear regression analysis
95 % Confidence interval
Variance inflation factors
C reactive protein (mg/dl)
Change of voriconazole concentration and clearance about the patient with and without glucocorticoid administration
mPSL 62.5 mg bid
DEX 20 mg
PSL 20 mg
Daily dose (mg/day)
Trough concentration (μg/ml)
Cutoff values of factors causing toxic level of voriconazole concentration
The results of the present study have shown that CRP elevation and hypoalbuminemia decreased voriconazole clearance and that concomitant glucocorticoid administration increased voriconazole clearance. In multiple linear regression analysis, we did not detect the presence of multicolinearity between CRP and glucocorticoid administration (variance inflation factors < 2), suggesting that CRP and concomitant glucocorticoid administration should be independent of each other in affecting voriconazole clearance. Although these two factors are independent, we speculate these factors might be associated with alteration of metabolism of voriconazole; elimination of voriconazole is mainly by metabolism of CYP2C19 dominantly and of CYP3A4 and CYP2C9 being involved to a much lesser extent [1–4].
Michael et al. retrospectively investigated therapeutic drug monitoring of voriconazole in a cohort of 201 patients . They identified by multiple linear regression analysis that the use of a systemic glucocorticoid was associated with significantly reduced voriconazole blood concentrations. They hypothesized that glucocorticoid up-regulated CYP2C19 and CYP3A4 expression since some in vitro [16, 17] and in vivo [18, 19] studies have shown that glucocorticoid can bind to and activate the pregnane X receptor, one of the nuclear receptors which plays a role in inducing cytochrome P450 expression, including CYP2C19 and CYP3A4, resulting in the increase of metabolism by the enzymes.
Another study has shown that voriconazole clearance significantly decreased in a limited number of patients with CYP2C19 poor metabolizer and heterozygous extensive metabolizer when coadministered with erythromycin, a known CYP3A4 inhibitor . In our study, of the three patients who administered concomitant glucocorticoid, C/D-ratio of one patient was considerably increased by discontinuation of glucocorticoid (Table 3, Case 1), and C/D-ratios of the two patients were considerably decreased by start of glucocorticoid (Table 3, Cases 2 and 3). They might be poor metabolizer of CYP2C19, resulting in the prominent increase of voriconazole metabolism due to CYP3A4 induction, although we did not perform the allele analysis of any gene of the patients.
Zordoky et al. reported that nuclear factor-κB plays a crucial role in the regulation of cytochrome P450 through several mechanisms and this role can explain the altered cytochrome P450 regulation in many conditions . They suggested that in inflammation status, nuclear factor-κB-repressing factor is activated and cytochrome P450 expression is decreased. A significant decrease in voriconazole clearance observed in patients with elevated CRP level in our study. Other studies reported the inflammation is associated with higher voriconazole trough concentrations [13, 21], congruent with our results. Therefore, higher voriconazole trough concentrations in inflammatory patients might be related to the above mechanism, since elevated CRP level and hypoalbuminemia are regarded as markers for inflammatory status.
In addition, we undertook an ROC analysis for prediction of risk of toxic voriconazole level on the basis of CRP and Alb. The results suggest that CRP and Alb should be useful for screening of toxic voriconazole level. In particular, in most cases, voriconazole concentrations reached toxic levels when the CRP and Alb values of the patients were over the cutoff values described below (Fig. 3c). Consequently, careful monitoring of voriconazole concentration and side effects are required for patients with both elevated CRP level and hypoalbuminemia. From our analysis, the cutoff values of CRP and Alb were 4.7 and 2.7 respectively. Statistically, these cutoff values are useful for prediction of toxic voriconazole level, although the value of 4.7 of CRP is less clinically important, since the value is common level of infectious diseases. Voriconazole concentrations in patients with hepatotoxicity were significantly higher than those without. In addition, the concentrations of voriconazole in 75 % of patients with hepatotoxicity were over 4 μg/ml. This result suggests that voriconazole trough concentration over 4 μg/ml should be avoided, supported by previous reports [8, 10].
A limitation of the present study is that CYP2C19 polymorphism was not available. However, Geist et al. found an enormous (88 %) interpatient variability in voriconazole trough concentrations, although none of the patients were classified as a CYP2C19 poor metabolizer. They noted that the high variability resulted from contributing factors, including changes in drug absorption, co-medication, and disease status . Further studies are needed to clarify contributing factors in patients with known genetic polymorphisms of the cytochrome P450 isoenzymes.
We evaluated patient-specific factors affecting voriconazole clearance and showed that increased CRP level and decreased Alb level were significant predictors of decreased voriconazole clearance, and coadministration of glucocorticoid was found to be a significant predictor of increased voriconazole clearance. It is not practical to examine CYP2C19 alleles due to the cost, so it is difficult to consider CYP2C19 polymorphism in therapeutic drug monitoring in routine practice. Therefore, we propose that early measurement of voriconazole concentration before the plateau phase should lead to the avoidance of toxic voriconazole level in patients with elevated CRP level and hypoalbuminemia.
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
- Dolton MJ, McLachlan AJ. Optimizing azole antifungal therapy in the prophylaxis and treatment of fungal infections. Curr Opin Infect Dis. 2014;27:493–500.View ArticlePubMedGoogle Scholar
- Traunmuller F, Popovic M, Konz KH, Smolle-Juttner FM, Joukhadar C. Efficacy and safety of current drug therapies for invasive aspergillosis. Pharmacology. 2011;88:213–24.View ArticlePubMedGoogle Scholar
- Seyedmousavi S, Mouton JW, Verweij PE, Bruggemann RJ. Therapeutic drug monitoring of voriconazole and posaconazole for invasive aspergillosis. Expert Rev Anti Infect Ther. 2013;11:931–41.View ArticlePubMedGoogle Scholar
- Hamada Y, Tokimatsu I, Mikamo H, Kimura M, Seki M, Takakura S, Ohmagari N, Takahashi Y, Kasahara K, Matsumoto K, et al. Practice guidelines for therapeutic drug monitoring of voriconazole: a consensus review of the Japanese Society of Chemotherapy and the Japanese Society of Therapeutic Drug Monitoring. J Infect Chemother. 2013;19:381–92.View ArticlePubMedPubMed CentralGoogle Scholar
- Lee S, Kim BH, Nam WS, Yoon SH, Cho JY, Shin SG, Jang IJ, Yu KS. Effect of CYP2C19 polymorphism on the pharmacokinetics of voriconazole after single and multiple doses in healthy volunteers. J Clin Pharmacol. 2012;52:195–203.View ArticlePubMedGoogle Scholar
- Shimizu T, Ochiai H, Asell F, Shimizu H, Saitoh R, Hama Y, Katada J, Hashimoto M, Matsui H, Taki K, et al. Bioinformatics research on inter-racial difference in drug metabolism I. Analysis on frequencies of mutant alleles and poor metabolizers on CYP2D6 and CYP2C19. Drug Metab Pharmacokinet. 2003;18:48–70.View ArticlePubMedGoogle Scholar
- Ueda K, Nannya Y, Kumano K, Hangaishi A, Takahashi T, Imai Y, Kurokawa M. Monitoring trough concentration of voriconazole is important to ensure successful antifungal therapy and to avoid hepatic damage in patients with hematological disorders. Int J Hematol. 2009;89:592–9.View ArticlePubMedGoogle Scholar
- Hamada Y, Seto Y, Yago K, Kuroyama M. Investigation and threshold of optimum blood concentration of voriconazole: a descriptive statistical meta-analysis. J Infect Chemother. 2012;18:501–7.View ArticlePubMedGoogle Scholar
- Dolton MJ, Ray JE, Chen SC, Ng K, Pont LG, McLachlan AJ. Multicenter study of voriconazole pharmacokinetics and therapeutic drug monitoring. Antimicrob Agents Chemother. 2012;56:4793–9.View ArticlePubMedPubMed CentralGoogle Scholar
- Suzuki Y, Tokimatsu I, Sato Y, Kawasaki K, Goto T, Hashinaga K, Itoh H, Hiramatsu K, Kadota J. Association of sustained high plasma trough concentration of voriconazole with the incidence of hepatotoxicity. Clin Chim Acta. 2013;424:119–22.View ArticlePubMedGoogle Scholar
- Cockcroft DW, Gault MH. Prediction of creatinine clearance from serum creatinine. Nephron. 1976;16:31–41.View ArticlePubMedGoogle Scholar
- Shi HY, Yan J, Zhu WH, Yang GP, Tan ZR, Wu WH, Zhou G, Chen XP, Ouyang DS. Effects of erythromycin on voriconazole pharmacokinetics and association with CYP2C19 polymorphism. Eur J Clin Pharmacol. 2010;66:1131–6.View ArticlePubMedPubMed CentralGoogle Scholar
- Naito T, Yamada T, Mino Y, Kawakami J. Impact of inflammation and concomitant glucocorticoid administration on plasma concentration of triazole antifungals in immunocompromised patients. Clin Chim Acta. 2015;441:127–32.View ArticlePubMedGoogle Scholar
- Usuki H, Inoue T, Ishida T, Miyazaki N. Determination of voriconazole by high-performance liquid chromatography. Igaku To Yakugaku. 2005;53:817–21.Google Scholar
- VFEND® Tablets [package insert]. Pfizer Japan Inc., Tokyo, Japan; Revised October. 2014.
- Chen Y, Ferguson SS, Negishi M, Goldstein JA. Identification of constitutive androstane receptor and glucocorticoid receptor binding sites in the CYP2C19 promoter. Mol Pharmacol. 2003;64:316–24.View ArticlePubMedGoogle Scholar
- Martin P, Riley R, Back DJ, Owen A. Comparison of the induction profile for drug disposition proteins by typical nuclear receptor activators in human hepatic and intestinal cells. Br J Pharmacol. 2008;153:805–19.View ArticlePubMedPubMed CentralGoogle Scholar
- Villikka K, Varis T, Backman JT, Neuvonen PJ, Kivisto KT. Effect of methylprednisolone on CYP3A4-mediated drug metabolism in vivo. Eur J Clin Pharmacol. 2001;57:457–60.View ArticlePubMedGoogle Scholar
- Shimada T, Terada A, Yokogawa K, Kaneko H, Nomura M, Kaji K, Kaneko S, Kobayashi K, Miyamoto K. Lowered blood concentration of tacrolimus and its recovery with changes in expression of CYP3A and P-glycoprotein after high-dose steroid therapy. Transplantation. 2002;74:1419–24.View ArticlePubMedGoogle Scholar
- Zordoky BN, El-Kadi AO. Role of NF-kappaB in the regulation of cytochrome P450 enzymes. Curr Drug Metab. 2009;10:164–78.View ArticlePubMedGoogle Scholar
- van Wanrooy MJ, Span LF, Rodgers MG, van den Heuvel ER, Uges DR, van der Werf TS, Kosterink JG, Alffenaar JW. Inflammation is associated with voriconazole trough concentrations. Antimicrob Agents Chemother. 2014;58:7098–101.View ArticlePubMedPubMed CentralGoogle Scholar
- Geist MJ, Egerer G, Burhenne J, Riedel KD, Weiss J, Mikus G. Steady-state pharmacokinetics and metabolism of voriconazole in patients. J Antimicrob Chemother. 2013;68:2592–9.View ArticlePubMedGoogle Scholar