Effects of new 17α-hydroxylase/C17,20-lyase inhibitors on LNCaP prostate cancer cell growth in vitro and in vivo

D. N. Grigoryev, B. J. Long, I. P. Nnane, V. C O Njar, Y. Liu, A. M H Brodie

Research output: Contribution to journalArticle

Abstract

Our laboratory has been developing new inhibitors of a key regulatory enzyme of testicular and adrenal androgen synthesis 17α-hydroxylase/C17,20-lyase (P450c17), with the aim of improving prostate cancer treatment. We designed and evaluated two groups of azolyl steroids: Δ5-non-competitive inhibitors (Δ5NCIs), VN/63-1, VN/85-1, VN/87-1 and their corresponding Δ4 derivatives (Δ4NCIs), VN/107-1, VN.108-1 and VN/109-1. The human P450c17 gene was transfected into LNCaP human prostate cancer cells, and the resultant LNCaP-CYP17 cells were utilized to evaluate the inhibitory potency of the new azolyl steroids. VN/85-1 and VN/108-1 had the lowest IC50 values of 1.25 ± 0.44 nM and 2.96 ± 0.78 nM respectively, which are much lower than that of the known P450 inhibitor ketoconazole (80.7 ± 1.8 nM). To determine whether the compounds had direct actions on proliferation of wild-type LNCaP cells, cell growth studies were performed. All of the Δ5NCIs and VN/108-1 blocked the growth-stimulating effects of androgens. In steroid-free media, the Δ5NCIs decreased the proliferation of LNCaP cells by 35-40%, while all of the Δ4NCIs stimulated LNCaP cells growth 1.5- to 2-fold. In androgen receptor (AR) binding studies, carried out to determine the mechanism of this effect, all of the Δ4NCIs (5 μM) displaced 77-82% of synthetic androgen R1881 (5 nM) from the LNCaP AR. The anti-androgen flutamide and the Δ5NCIs displaced 53% and 32-51% of R1881 bound to AR respectively. These results suggested that the Δ5NCIs may also be acting as anti-androgens. We further evaluated our inhibitors in male severe combined immunodeficient mice bearing LNCaP tumour xenografts. In this model VN/85-1 was as effective as finasteride at inhibiting tumor growth (26% and 28% inhibition, respectively) and the inhibitory effect of VN/87-1 was similar to that of castration (33% and 36% inhibition respectively). These results suggest that VN/85-1 and VN/87-1 may be potential candidates for treatment of prostate cancer.

Original languageEnglish (US)
Pages (from-to)622-630
Number of pages9
JournalBritish Journal of Cancer
Volume81
Issue number4
DOIs
StatePublished - 1999
Externally publishedYes

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Lyases
Mixed Function Oxygenases
Prostatic Neoplasms
Androgens
Androgen Receptors
Metribolone
Growth
Steroids
Testosterone Congeners
Steroid 17-alpha-Hydroxylase
Finasteride
Flutamide
Ketoconazole
SCID Mice
Castration
Heterografts
Inhibitory Concentration 50
Neoplasms
Cell Proliferation
In Vitro Techniques

Keywords

  • 17α-hydroxylase/C-lyase inhibitors
  • Androgens synthesis
  • Azolyl steroids
  • LNCaP cells
  • SCID mice

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Effects of new 17α-hydroxylase/C17,20-lyase inhibitors on LNCaP prostate cancer cell growth in vitro and in vivo. / Grigoryev, D. N.; Long, B. J.; Nnane, I. P.; Njar, V. C O; Liu, Y.; Brodie, A. M H.

In: British Journal of Cancer, Vol. 81, No. 4, 1999, p. 622-630.

Research output: Contribution to journalArticle

Grigoryev, D. N. ; Long, B. J. ; Nnane, I. P. ; Njar, V. C O ; Liu, Y. ; Brodie, A. M H. / Effects of new 17α-hydroxylase/C17,20-lyase inhibitors on LNCaP prostate cancer cell growth in vitro and in vivo. In: British Journal of Cancer. 1999 ; Vol. 81, No. 4. pp. 622-630.
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abstract = "Our laboratory has been developing new inhibitors of a key regulatory enzyme of testicular and adrenal androgen synthesis 17α-hydroxylase/C17,20-lyase (P450c17), with the aim of improving prostate cancer treatment. We designed and evaluated two groups of azolyl steroids: Δ5-non-competitive inhibitors (Δ5NCIs), VN/63-1, VN/85-1, VN/87-1 and their corresponding Δ4 derivatives (Δ4NCIs), VN/107-1, VN.108-1 and VN/109-1. The human P450c17 gene was transfected into LNCaP human prostate cancer cells, and the resultant LNCaP-CYP17 cells were utilized to evaluate the inhibitory potency of the new azolyl steroids. VN/85-1 and VN/108-1 had the lowest IC50 values of 1.25 ± 0.44 nM and 2.96 ± 0.78 nM respectively, which are much lower than that of the known P450 inhibitor ketoconazole (80.7 ± 1.8 nM). To determine whether the compounds had direct actions on proliferation of wild-type LNCaP cells, cell growth studies were performed. All of the Δ5NCIs and VN/108-1 blocked the growth-stimulating effects of androgens. In steroid-free media, the Δ5NCIs decreased the proliferation of LNCaP cells by 35-40{\%}, while all of the Δ4NCIs stimulated LNCaP cells growth 1.5- to 2-fold. In androgen receptor (AR) binding studies, carried out to determine the mechanism of this effect, all of the Δ4NCIs (5 μM) displaced 77-82{\%} of synthetic androgen R1881 (5 nM) from the LNCaP AR. The anti-androgen flutamide and the Δ5NCIs displaced 53{\%} and 32-51{\%} of R1881 bound to AR respectively. These results suggested that the Δ5NCIs may also be acting as anti-androgens. We further evaluated our inhibitors in male severe combined immunodeficient mice bearing LNCaP tumour xenografts. In this model VN/85-1 was as effective as finasteride at inhibiting tumor growth (26{\%} and 28{\%} inhibition, respectively) and the inhibitory effect of VN/87-1 was similar to that of castration (33{\%} and 36{\%} inhibition respectively). These results suggest that VN/85-1 and VN/87-1 may be potential candidates for treatment of prostate cancer.",
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AU - Long, B. J.

AU - Nnane, I. P.

AU - Njar, V. C O

AU - Liu, Y.

AU - Brodie, A. M H

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