Biochemical and Biophysical Research Communications 477 (2016) 1005e1010
`
`Contents lists available at ScienceDirect
`
`Biochemical and Biophysical Research Communications
`
`j o u r n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e / y b b r c
`
`Specificity of anti-prostate cancer CYP17A1 inhibitors on androgen
`biosynthesis
`
`Sameer S. Udhane a, b, Bernhard Dick b, c, Qingzhong Hu d, 1, Rolf W. Hartmann d, e,
`Amit V. Pandey a, b, *
`
`a Pediatric Endocrinology, Diabetology and Metabolism, Department of Pediatrics, University Children’s Hospital Bern, 3010 Bern, Switzerland
`b Department of Clinical Research, University of Bern, 3010 Bern, Switzerland
`c Department of Nephrology, Hypertension and Clinical Pharmacology, University Hospital of Bern, Bern, Switzerland
`d Pharmaceutical and Medicinal Chemistry, Saarland University, Campus C2.3, Saarbrücken, Germany
`e Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Campus E 8.1, 66123 Saarbrücken, Germany
`
`a r t i c l e i n f o
`
`a b s t r a c t
`
`Article history:
`Received 26 June 2016
`Accepted 4 July 2016
`Available online 6 July 2016
`
`Keywords:
`CYP17A1
`Orteronel
`Abiraterone
`CYP21A2
`PCOS
`Premature adrenarche
`Castration resistant prostate cancer
`
`The orteronel, abiraterone and galeterone, which were developed to treat castration resistant prostate
`cancer, inhibit 17,20 lyase activity but little is known about their effects on adrenal androgen biosyn-
`thesis. We studied the effect of several inhibitors and found that orteronel was selective towards 17,20
`lyase activity than abiraterone and galeterone. Gene expression analysis showed that galeterone altered
`the expression of HSD3B2 but orteronel did not change the expression of HSD3B2, CYP17A1 and AKR1C3.
`The CYP19A1 activity was not inhibited except by compound IV which lowered activity by 23%. Sur-
`prisingly abiraterone caused complete blockade of CYP21A2 activity. Analysis of steroid metabolome by
`gas chromatography e mass spectrometry revealed changes in steroid levels caused by different in-
`hibitors. We can conclude that orteronel is a highly specific inhibitor of 17,20 lyase activity. The discovery
`of these specific drug actions on steroidogenic enzyme activities would be valuable for understanding
`the regulation of androgens.
`
`© 2016 Elsevier Inc. All rights reserved.
`
`1. Introduction
`
`Androgens are essential for sexual differentiation and repro-
`duction in both male and female. Androgen production in humans
`occurs mainly in the testis, ovaries and the adrenal glands. The
`initial steps of steroidogenesis in both the adrenal and the gonads
`are same and use similar enzymes for steroid biosynthesis path-
`ways [1]. Both the adrenal and the ovary produce dehydroepian-
`drosterone (DHEA) as the key precursor of androgens and
`estrogens. Premature adrenarche and polycystic ovary syndrome
`(PCOS) are common hyperandrogenic disorders. During adrenarche
`(functional activation of adrenal zona reticularis) production of
`adrenal androgens (DHEA and DHEA sulfate) is at a higher level [1].
`
`* Corresponding author. Pediatric Endocrinology, Diabetology & Metabolism,
`KIKL C837, University Children’s Hospital Bern, Freiburgstrasse 15, CH-3010 Bern,
`Switzerland.
`E-mail address: amit@pandeylab.org (A.V. Pandey).
`1 Current address: Department of Chemistry, University of Cambridge, Lensfield
`Road, Cambridge, CB2 1EW, U.K.
`
`http://dx.doi.org/10.1016/j.bbrc.2016.07.019
`0006-291X/© 2016 Elsevier Inc. All rights reserved.
`
`In case of premature adrenarche, the concentration of circulating
`adrenal androgens increases at an earlier age. Several studies link
`premature adrenarche with post pubertal adrenal and ovarian
`hyperandrogenism [2,3]. Premature adrenarche is also linked to the
`hyperandrogenism in PCOS [4]. In hyperandrogenic women with
`PCOS, androgen production from both the ovaries and the adrenals
`is higher [5]. Changes in cytochrome P450 17A1 (P450c17,
`CYP17A1) activities could explain the increase in androgen secre-
`tion that causes hyperandrogenic disorders [1].
`The CYP17A1 plays a vital role in regulating adrenal androgen
`production (Suppl Fig. 1). CYP17A1 is localized in the endoplasmic
`reticulum and responsible for 17a-hydroxylase and 17,20 lyase re-
`actions. The 17,20 lyase activity regulates androgen production and
`is dependent on cytochrome P450 oxidoreductase (POR) [6], cyto-
`chrome b5 (CYB5) [7] and phosphorylation [7,8]. Treatment of
`androgen dependent prostate cancer is by androgen deprivation
`therapy, androgen receptor (AR) antagonists or anti androgen
`agents and CYP17A1 inhibitors that block androgen production [9].
`Hormonal therapy such as using a gonadotrophin hormonal
`analogue for treatment of hyperandrogenic disorders may not be
`
`Amerigen Exhibit 1107
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`specific enough to control the extra-gonadal and the gonadal
`androgen production. Drugs targeting androgen production are
`being used for treatment of prostate cancer and to decrease the
`androgen levels in hyperandrogenic women [10,11]. But anti
`androgen treatment can have side effects on sexuality of women
`with hyperandrogenism [12]. One example is flutamide, a non-
`steroidal anti androgen drug used to treat hyperandrogenic
`women [13e16]. Flutamide has many side effects like diarrhea,
`gynecomastia, muscle cramps, hematological alterations, and
`hepatotoxicity [17,18].
`CYP17A1 inhibitors are designed for the treatment of the
`androgen dependent prostate cancer patients (Suppl Fig. 2). Abir-
`aterone prolonged the survival of castration-resistant prostate
`cancer (CRPC) patients in phase III clinical trials [19]. Abiraterone
`also has modest affinity towards the androgen receptor [20]. In the
`hyperandrogenic disorders like premature or exaggerated adre-
`narche and PCOS, use of total CYP17A1 inhibitors will also block the
`17a-hydroxylase activity. New inhibitors of CYP17A1, orteronel and
`galeterone were also designed for treatment of castration-resistant
`prostate cancer [21,22]. Effect of abiraterone, orteronel and gale-
`terone on adrenal androgen production is not clear.
`Here we are reporting the effects of CYP17A1 inhibitors on the
`adrenal androgen production. We compared CYP17A1 inhibitors
`orteronel, galeterone, abiraterone and recently reported com-
`pounds I and IV and studied the specificity of their action. We found
`that orteronel is a potent inhibitor of 17,20 lyase activity with no
`impact on other cytochrome P450 enzymes involved in steroid
`hormone biosynthesis. We also analyzed the impact of these in-
`hibitors on the steroid metabolome, focusing on C19 steroids. Use
`of specific 17,20 lyase inhibitors can be a treatment option for
`hyperandrogenic disorders for the control of androgen production.
`
`2. Materials and methods
`
`2.1. Cell culture and treatment
`
`Human adrenocortical NCI-H295R cells obtained from American
`Type Culture Collection (ATCC; CRL-2128) were cultivated in
`DMEM/Ham’s F-12 medium with additives as described in
`supplementary material. Orteronel, galeterone and abiraterone
`were purchased from the Selleckchem (Houston, TX, USA).
`CYP17A1 inhibitors compound I and IV were described earlier [23].
`
`2.2. Steroid profiling
`
`Steroid profiling was performed with the cells grown on 6 well
`plates by adding 100,000 cpm [3H] pregnenolone (preg) to the
`culture medium for 90 min. To study specific CYP17A1 enzyme
`activities, cells were first treated with 1 mM or 2 mM CYP17A1 in-
`hibitors. After 24 h of inhibitor treatment, cells were treated with
`1 mM trilostane (a specific blocker of HSD3B) for 90 min before
`adding [3H] preg. For CYP21A2 activity, cells were treated with
`1 mM of different inhibitors for 24 h and then labeled with [3H] 17a-
`hydroxyprogesterone (17-OH Prog) for 60 min to monitor the
`conversion of 11-deoxycortisol. Steroids were extracted from cell
`supernatants and separated by thin layer chromatography (TLC) as
`previously described [24]. Steroid conversion was calculated as
`percentage of
`total
`radioactivity incorporated into specific
`products.
`
`2.3. Quantitative real time PCR (qRT-PCR)
`
`2.4. CYP19A1 enzyme activity using titrated water release assay
`
`Human Placental JEG3 cells were grown on 12 well plates and
`treated with CYP17A1 inhibitors for 24 h before aromatase activity
`was assayed. Aromatase activity was assayed with 3H labeled an-
`drostenedione ([1b-3H(N)]-androstene-3,17-dione; ~100,000 cpm/
`well). Androstenedione was added to the treated cells for 6 h.
`Aromatase activity was then assessed by the titrated water release
`assay as previously described [25,26].
`
`2.5. GC-MS measurement of steroid metabolites
`
`The measurement of steroid hormone metabolites in human
`adrenal H295R cells was performed by gas chromatography e mass
`spectrometry (GC-MS) as previously described [27]. The cells were
`grown in 10 cm plates in normal growth medium for 24 h, then
`medium was replaced, and cells were treated with 1 mM of CYP17A1
`inhibitors in medium without NU-I serum for 24 h. After 12 h of
`treatment, 1 mM of pregnenolone was added. At the end, superna-
`tant was collected and concentrated samples were used for steroid
`analysis by GCeMS. All measurements were performed in the
`steroid laboratory of the Department of Nephrology, Hypertension
`and Clinical Pharmacology at the University Hospital of Bern,
`Switzerland.
`
`3. Results and discussion
`
`3.1. CYP17A1 inhibitors modulate the adrenal androgen production
`
`CYP17A1 is required for androgen production in humans and
`control of its enzyme activity is essential for treatment of hyper-
`androgenic disorders. Abiraterone, galeterone and orteronel are
`inhibitors of CYP17A1 (Suppl Fig. 2). Abiraterone is an effective
`active site-directed inhibitor, designed for the treatment of CRPC
`patients to suppress the androgen production by inhibition of
`CYP17A1 activities [28,29]. The galeterone and orteronel were
`designed to selectively inhibit 17,20 lyase activity of CYP17A1 for
`the treatment of CRPC [21,22]. There have been suggestions that
`galeterone is more specific towards the 17,20 lyase activity of
`CYP17A1 than abiraterone but this effect is not clear [9,30]. Func-
`tional studies on the effects of orteronel in vivo (cynomolgus
`monkey) and in vitro (human adrenal tumor cells) showed that
`orteronel is a potent inhibitor of 17,20 lyase activity [31]. In addition
`to these well-known inhibitors, Hartmann group synthesized
`inhibitors of steroid biosynthesis [23]. They observed
`several
`compound IV was a more potent inhibitor of 17,20 lyase activity
`compared to its effects on 17a-hydroxylase activity [23]. However,
`the effect of these drugs on adrenal androgen production remains
`unknown.
`We used the steroidogenic human adrenal cortex H295R cells to
`study the effect of CYP17A1 inhibitors (orteronel, galeterone,
`compound I and IV and abiraterone). We found that orteronel and
`galeterone drastically decreased the DHEA and androstenedione
`production (Fig. 1A). Compounds I and IV decreased DHEA,
`androstendione and 11-DOC/11-deoxycortisol production. In the
`cells treated with abiraterone, we only observed the conversion of
`preg to prog and other products in the pathway were completely
`blocked (Fig. 1B). This suggests that abiraterone inhibits CYP17A1
`and possibly other enzymes that are involved in steroidogenesis.
`
`3.2. Orteronel is a specific inhibitor of the CYP17A1 17,20 lyase
`activity
`
`The H295R cells were treated with inhibitors for 72 h. After the
`treatment, total RNA was isolated and qRT-PCR was done as pre-
`viously described [24]. Details are in supplementary material.
`
`We studied the effect of CYP17A1 inhibitors in more detail for
`the specificity of the inhibition of CYP17A1 enzyme activities. Cells
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`1007
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`Fig. 1. Effect of the CYP17A1 inhibitors on adrenal steroid profile. For the steroid profiling H295R cells were treated with 1 mm orteronel, galeterone, compound I and IV and
`abiraterone for 24 h. Steroid production was labeled with [3H] preg for 90 min. Steroids were extracted and resolved by TLC. A) Steroid profiling of adrenal H295R cells treated with
`DMSO and 1 mM of orteronel or galeterone B) Steroid profiling of adrenal H295R cells treated with 1 mM of compound I, compound IV and abiraterone. Representative TLCs are
`depicted on the left, summaries and quantifications of the results are given on the right. Prog, progesterone; D4A, androstenedione; Preg, pregnenolone; DHEA, dehydroepian-
`drosterone; 17-OH Preg, 17a-hydroxypregnenolone; 17-OH Prog, 17a-hydroxy progesterone; Data are presented as mean ± SD of three independent experiments. *p < 0.05,
`**p < 0.01.
`
`Fig. 2. Effects of CYP17A1 inhibitors on NCI-H295 cells. A and B . Effect of inhibitors on CYP17A1 enzyme activity. The enzyme activities of CYP17A1 were checked in H295R
`cells treated with control (DMSO) and 1 mM of orteronel, galeterone, compound I, compound IV and abiraterone. CYP17A1 activities were assessed by the conversion of [3H] preg to
`17-OH preg for 17a-hydroxylase activity and 17-OH preg to DHEA for 17,20 lyase activity. Representative TLCs are depicted on the left, summaries and quantifications of the results
`are given on the right. Data are given as mean ± SD of three independent experiments. Preg, pregnenolone; DHEA, dehydroepiandrosterone; 17-OH Preg, 17a-hydroxypregnenolone.
`*p < 0.05,**p < 0.01. C. Impact of CYP17A1 inhibitors on genes involved in androgen biosynthesis. H295R cells were treated with control (DMSO) or 1 mM orteronel and
`galeterone for 72 h and then RNA was isolated and transcribed to cDNA. Results of qRT-PCR validation of HSD3B2, CYP17A1 and AKR1C3 genes relative to the housekeeping gene
`cyclophilin A. Expression of the genes was analyzed by SYBR Green real-time PCR. Analysis of relative gene expression was determined by the 2DDCt method. *p < 0.05.
`
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`were treated with inhibitors and then with 1 mM trilostane (a
`specific blocker of HSD3B) for 90 min before labeling with [3H]
`preg. We found that the orteronel decreased only DHEA production,
`suggesting selectivity of inhibition towards the 17,20 lyase activity
`of CYP17A1 (Fig. 2A). Treatments of galeterone, abiraterone, com-
`pound I and IV showed a decrease in the conversion from preg to
`17-OH preg and to DHEA which suggests these compounds
`inhibited both the hydroxylase as well as lyase activity of CYP17A1
`(Fig. 2AeB). These data indicate that orteronel is a specific inhibitor
`of 17,20 lyase activity, as observed in previous studies [31,32].
`
`3.3. Effect of CYP17A1 inhibitors on genes involved in androgen
`biosynthesis
`
`Abiraterone and galeterone are steroidal structures that also act
`as androgen receptor (AR) antagonist. Orteronel is a non-steroidal
`CYP17A1 inhibitor. Abiraterone has been reported to alter the
`androgen regulated gene expression in prostate cancer cells
`through the AR-mediated signaling [33]. Since, both the orteronel
`and galeterone have shown potential as potent CYP17A1 inhibitors,
`we conducted further experiments to find out if these inhibitors
`have any impact on gene transcription. The H295R cells were
`treated with control (DMSO) or 1 mM orteronel and galeterone for
`72 h. Gene expressions were studied by relative quantification PCR
`(qRT-PCR). We studied the expression of three key genes CYP17A1,
`HSD3B2 and AKR1C3 that are involved in androgen biosynthesis.
`Interestingly, we found a slight increase in HSD3B2 gene expression
`upon treatment with galeterone (Fig. 2C). In presence of orteronel
`no significant changes in gene expression were observed (Fig. 2C).
`
`3.4.
`
`Influence of CYP17A1 inhibitors on aromatase activity
`
`To further study the specificity of the CYP17A1 inhibitors, we
`determined their impact on other related steroidogenic enzyme
`activities. First, we studied the effect of inhibitors on aromatase
`(CYP19A1) activity. The activity and expression of aromatase differs
`between different cell types depending on the need of cells for
`estrogens. Aromatase is localized in endoplasmic reticulum and
`catalyzes the last step of estrogen biosynthesis from androgens
`(converts androstenedione to estrone and testosterone to estra-
`diol). The orteronel, galeterone, compound I and abiraterone
`showed no effect of on aromatase activity (Fig. 3A). Compound IV
`treatment resulted in a decrease of aromatase activity by 23%
`(Fig. 3A), which was in accordance with its reported modest inhi-
`bition of aromatase in vitro (IC50 ¼ 228 nM) [34].
`
`3.5. Abiraterone severely inhibits 21-hydroxylase (CYP21A2)
`activity
`
`We then studied the effect of these inhibitors on the 21-
`hydroxylase (CYP21A2) enzyme activity. The 21-hydroxylase plays
`a role in producing glucocorticoids (cortisol) and mineralocorticoid
`(aldosterone). Cortisol helps maintain blood sugar levels, protects
`the body from stress and suppresses inflammation. Maintaining the
`cortisol level is required for the gonadal function, as disruption in
`cortisol production may affect
`fertility by the regulation of
`hypothalamic-pituitary-adrenal (HPA) and gonadal (HPG) axis [35].
`CYP21A2 activities were assessed by checking the conversion of
`[3H] 17-OH Prog to 11-deoxycortisol. Orteronel, galeterone, com-
`pound I and IV treatment did not change 11-deoxycortisol pro-
`duction, suggesting no effect on CYP21A2 activity. Surprisingly,
`abiraterone was found to completely inhibit the CYP21A2 activity
`(Fig. 3B). This suggests that abiraterone is not a specific inhibitor of
`CYP17A1 and can also inhibit CYP21A2 activity. Protein sequence
`comparison of CYP17A1 and CYP21A2 shows an identity of around
`
`Fig. 3. Influence of CYP17A1 inhibitors on other steroid metabolizing cytochrome
`P450 activity. The effect of
`inhibitors on P450 aromatase (CYP19A1) and 21-
`hydroxylase (CYP21A2). A) Effect of inhibitors on aromatase. Effect of inhibitors
`was studied on aromatase activity in human placental JEG-3 cells. Cells were treated
`with control (DMSO) and 1 mM of orteronel, galeterone, abiraterone, compound I and
`IV for 24 h before aromatase activity was tested. Aromatase activity was assayed with
`androstenedione using 2 mM cold substrate and 3H labeled androstenedione ([1b-
`3H(N)]-androstene-3,17-dione; ~100,000 cpm/well) as radioactive tracer. Andro-
`stenedione was added to the treated cells for 6 h. Aromatase activity was measured by
`observing the conversion of 3H-androstenedione to estrone using the titrated water
`release assay. Data are presented as mean ± SEM of two independent experiments. B)
`Effect on CYP21A1 activity. The 21-hydroxylase activity was checked in H295R cells
`treated with control (DMSO) and 1 mM of orteronel, galeterone, abiraterone, compound
`I and compound IV for 24 h. CYP21A2 activities were measured by observing the
`conversion of [3H] 17-OH prog to 11-deoxycortisol. Representative TLCs are depicted in
`the upper panel, summaries and quantifications of the results are given in the lower
`panel. Data are presented as mean ± SD of three independent experiments. **p < 0.01.
`
`28% but structures of CYP17A1 and CYP21A2 have many similarities
`and may retain similar binding sites. The inhibition of CYP21A2
`could be a serious problem since using this drug inhibits whole
`adrenal androgen biosynthesis. Use of abiraterone for androgen
`lowering effects, where inhibition of CYP21A2 is not desired, will
`create additional complications.
`
`Impact of CYP17A1 inhibitors on the metabolomics of C19
`3.6.
`steroids by GC-MS
`
`The CYP17A1 inhibitors orteronel, galeterone and abiraterone
`showed significant decrease in potent androgens like testosterone
`and dihydrotestosterone (Table 1). Galeterone lowered all classical
`and backdoor pathway metabolites of androgens in human adrenal
`cells. However, orteronel and abiraterone treatment lead to an in-
`crease of androgen metabolite androsterone, which is involved in
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`
`Table 1
`The table shows the GC-MS measurement of secreted steroid metabolites in the human adrenal H295R cells in the untreated vs the treated condition. The change in me-
`tabolites are indicated based on untreated levels which are set as hundred percent compared to CYP17A1 inhibitor treatments. *nd: Not determined; þ: Increased metabolite.
`
`Metabolites
`
`Untreated
`
`Orteronel
`
`Galeterone
`
`Abiraterone
`
`Androgen metabolites
`Androsterone
`Etiocholanolone
`Androstenediol
`11b-Hydroxy-Androsterone
`11b-Hydroxy-Etiocholanolone
`Dehydroepiandrosterone
`5a androstendiol
`16a-Hydroxy-DHEA
`5a androstenetriol
`5-Pregnenetriol
`Testosterone
`5a-Dihydrotestosterone
`Estrogen metabolites
`Estriol
`17-b Estradiol
`Progesterone metabolites
`17a-OH-Pregnanolone
`Pregnanetriol
`Corticosterone metabolites
`Tetrahydrocorticosterone
`5a-Tetrahydrocorticosterone
`Cortisol-metabolites
`Cortisone
`Tetrahydrocortisone
`a-Cortolone
`b-Cortolone
`20a-Dihydrocortisone
`20b-Dihydrocortisone
`Cortisol
`TH-Cortisol
`5a-Tetrahydrocortisol
`a-Cortol
`b-Cortol
`20a-Dihydrocortisol
`
`100
`100
`100
`100
`nd
`100
`100
`100
`100
`100
`100
`100
`
`100
`nd
`
`100
`100
`
`nd
`nd
`
`nd
`100
`100
`nd
`nd
`nd
`100
`100
`100
`nd
`nd
`nd
`
`448
`3915
`328
`104
`
`þ
`48
`80
`74
`108
`36
`50
`nd
`
`103
`
`þ
`
`4162
`802
`
`nd
`nd
`
`nd
`90
`873
`
`þ
`nd
`nd
`22
`nd
`nd
`
`þ
`nd
`120
`
`30
`51
`nd
`94
`nd
`30
`66
`84
`39
`71
`29
`nd
`
`58
`
`þ
`
`56
`49
`
`þ
`nd
`
`nd
`96
`374
`
`þ
`nd
`nd
`22
`83
`83
`
`þ
`
`þ
`162
`
`1593
`3030
`152
`1834
`
`þ
`10
`63
`81
`133
`9
`26
`nd
`
`55
`
`þ
`
`4408
`2011
`
`þ
`
`þ
`
`þ
`6372
`45,600
`
`þ
`
`þ
`
`þ
`40
`2617
`2617
`
`þ
`
`þ
`161
`
`the backdoor pathway of dihydrotestosterone biosynthesis. One
`possible explanation is that abiraterone can influence backdoor
`pathway by elevating CYP17A1 expression and induce progesterone
`accumulation [36]. The progesterone accumulation can compete
`against abiraterone for binding to CYP17A1 leading to induction of
`androgen metabolites involved in backdoor pathway [36]. Orter-
`onel seems to be more specific for inhibiting the 17,20 lyase activity
`in the classical androgen biosynthesis pathway and might not in-
`fluence the androgen metabolism via backdoor pathway. Addi-
`tionally, we found that orteronel and abiraterone treatment also
`lead to an increase in other androgen metabolites like etiochola-
`nolone and 11b-hydroxy-etiocholanolone. For the first time the
`data here show the effect of CYP17A1 inhibitors on the human
`steroid metabolome especially on androgen metabolites, in adrenal
`H295R cells. These data indicate that orteronel, galeterone and
`abiraterone can influence or alter different steroid metabolites in
`human adrenal cells apart from blocking the CYP17A1 activity.
`Based on our results, we can conclude that orteronel is a more
`specific inhibitor for CYP17A1 17,20 lyase activity compared to other
`inhibitors tested here. Orteronel had no effect on other tested en-
`zymes that are involved in steroid hormone biosynthesis. Hence, it
`can be considered as a treatment option for hyperandrogenic dis-
`orders like polycystic ovary syndrome. Using abiraterone for
`lowering androgen production in hyperandrogenic disorders may
`create additional undesired complications in patients because of its
`inhibitory effect on CYP21A2 activities. For the first time we
`showed the effect of CYP17A1 inhibitors abiraterone, orteronel and
`galeterone on steroid metabolome in human adrenal H295R cells.
`Our data provide evidence for the significantly altered steroids
`
`metabolites by CYP17A1 inhibitors. The steroid metabolome model
`may be used as an efficient diagnostic tool for further studies on
`urine or serum samples of hyperandrogenic patients treated with
`CYP17A1 inhibitors. Our observations that abiraterone is also a very
`potent inhibitor of CYP21A2, suggests that abiraterone should not
`be used to treat hyperandrogenic disorders when inhibition of
`CYP21A2 is an undesirable side effect. Orteronel may be tried for a
`more specific inhibition of CYP17A1 activities without affecting
`other steroid metabolizing enzymes.
`
`Acknowledgments
`
`This work was supported by grants to AVP from the Swiss Na-
`tional Science Foundation (31003A-134926). We thank Dr. Nasser
`Dhayat from the Department of Nephrology, Hypertension and
`Clinical Pharmacology, University Hospital of Bern, Bern,
`Switzerland for help with the steroid pathway figure.
`
`Appendix A. Supplementary data
`
`Supplementary data related to this article can be found at http://
`dx.doi.org/10.1016/j.bbrc.2016.07.019.
`
`Transparency document
`
`Transparency document related to this article can be found
`online at http://dx.doi.org/10.1016/j.bbrc.2016.07.019.
`
`

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`S.S. Udhane et al. / Biochemical and Biophysical Research Communications 477 (2016) 1005e1010
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