High expression of the androgen receptor (AR) and the disruption of its regulation are strongly responsible for the development of prostate cancer (PCa). Therapeutically relevant non-steroidal or steroidal antiandrogens are able to block the AR effect by eliminating AR-mediated signalling. Herein we report the synthesis of novel steroidal pyrazoles derived from the natural sex hormone 5α-dihydrotestosterone (DHT). 2-Ethylidene or 2-(hetero)arylidene derivatives of DHT obtained by regioselective Claisen-Schmidt condensation with acetaldehyde or (hetero)aromatic aldehydes in alkaline ethanol were reacted with monosubstituted hydrazines to give A-ring-fused 1,5-disubstituted pyrazoles as main or exclusive products, depending on the reaction conditions applied. Spontaneous or 2,3-dichloro-5,6-dicyanobenzoquinone (DDQ)-induced oxidation of the primarily formed pyrazolines resulted in the desired products in moderate to good yields, while 17-oxidation also occurred by using the Jones reagent as a strong oxidant. Transcriptional activity of the AR in a reporter cell line was examined for all novel compounds, and several previously synthesized similar DHT-based pyrazoles with differently substituted heteroring were also included to obtain information about the structure-activity relationship. Two specific regioisomeric groups of derivatives significantly diminished the transcriptional activity of the AR in reporter cell line in 10 μM concentration, and displayed reasonable antiproliferative activity in AR-positive PCa cell lines. Lead compound (3d) was found to be a potent AR antagonist (IC50 = 1.18 μM), it generally suppressed AR signalling in time and dose dependent manner, moreover, it also led to a sharp decrease in wt-AR protein level probably caused by proteasomal degradation. We confirmed the antiproliferative activity of 3d in AR-positive PCa cell lines (with GI50 in low micromolar ranges), and its cellular, biochemical and in silico binding in AR ligand-binding domain. Moreover, compound 3d was shown to be potent even ex vivo in patient-derived tissues, which highlights the therapeutic potential of A-ring-fused pyrazoles.

Department of Clinical and Molecular Pathology Institute of Molecular and Translational Medicine Palacký University Olomouc and University Hospital Olomouc Hněvotínská 3 77515 Olomouc Czech Republic

Department of Experimental Biology Faculty of Science Palacký University Olomouc Šlechtitelů 27 78371 Olomouc Czech Republic

Department of Organic Chemistry University of Szeged Dóm Tér 8 H 6720 Szeged Hungary

Department of Urology Palacký University Olomouc and University Hospital Olomouc 1 P Pavlova 6 77900 Olomouc Czech Republic

000      

00000naa a2200000 a 4500

001      

bmc23003875

003      

CZ-PrNML

005      

20230425140929.0

007      

ta

008      

230418s2023 fr f 000 0|eng||

009      

AR

024    7_

$a 10.1016/j.ejmech.2023.115086 $2 doi

035    __

$a (PubMed)36682291

040    __

$a ABA008 $b cze $d ABA008 $e AACR2

041    0_

$a eng

044    __

$a fr

100    1_

$a Peřina, Miroslav $u Department of Experimental Biology, Faculty of Science, Palacký University Olomouc, Šlechtitelů 27, 78371, Olomouc, Czech Republic

245    10

$a A-ring-fused pyrazoles of dihydrotestosterone targeting prostate cancer cells via the downregulation of the androgen receptor / $c M. Peřina, MA. Kiss, G. Mótyán, E. Szczyrbová, M. Eliáš, V. Študent, D. Kurfürstová, M. Kovalová, L. Mada, J. Bouchal, É. Frank, R. Jorda

520    9_

$a High expression of the androgen receptor (AR) and the disruption of its regulation are strongly responsible for the development of prostate cancer (PCa). Therapeutically relevant non-steroidal or steroidal antiandrogens are able to block the AR effect by eliminating AR-mediated signalling. Herein we report the synthesis of novel steroidal pyrazoles derived from the natural sex hormone 5α-dihydrotestosterone (DHT). 2-Ethylidene or 2-(hetero)arylidene derivatives of DHT obtained by regioselective Claisen-Schmidt condensation with acetaldehyde or (hetero)aromatic aldehydes in alkaline ethanol were reacted with monosubstituted hydrazines to give A-ring-fused 1,5-disubstituted pyrazoles as main or exclusive products, depending on the reaction conditions applied. Spontaneous or 2,3-dichloro-5,6-dicyanobenzoquinone (DDQ)-induced oxidation of the primarily formed pyrazolines resulted in the desired products in moderate to good yields, while 17-oxidation also occurred by using the Jones reagent as a strong oxidant. Transcriptional activity of the AR in a reporter cell line was examined for all novel compounds, and several previously synthesized similar DHT-based pyrazoles with differently substituted heteroring were also included to obtain information about the structure-activity relationship. Two specific regioisomeric groups of derivatives significantly diminished the transcriptional activity of the AR in reporter cell line in 10 μM concentration, and displayed reasonable antiproliferative activity in AR-positive PCa cell lines. Lead compound (3d) was found to be a potent AR antagonist (IC50 = 1.18 μM), it generally suppressed AR signalling in time and dose dependent manner, moreover, it also led to a sharp decrease in wt-AR protein level probably caused by proteasomal degradation. We confirmed the antiproliferative activity of 3d in AR-positive PCa cell lines (with GI50 in low micromolar ranges), and its cellular, biochemical and in silico binding in AR ligand-binding domain. Moreover, compound 3d was shown to be potent even ex vivo in patient-derived tissues, which highlights the therapeutic potential of A-ring-fused pyrazoles.

650    _2

$a mužské pohlaví $7 D008297

650    _2

$a lidé $7 D006801

650    12

$a dihydrotestosteron $x farmakologie $x metabolismus $7 D013196

650    _2

$a androgenní receptory $x metabolismus $7 D011944

650    _2

$a pyrazoly $7 D011720

650    _2

$a down regulace $7 D015536

650    _2

$a nádorové buněčné linie $7 D045744

650    12

$a nádory prostaty $x farmakoterapie $x metabolismus $7 D011471

650    _2

$a steroidy $x terapeutické užití $7 D013256

655    _2

$a časopisecké články $7 D016428

700    1_

$a Kiss, Márton A $u Department of Organic Chemistry, University of Szeged, Dóm Tér 8, H-6720, Szeged, Hungary

700    1_

$a Mótyán, Gergő $u Department of Organic Chemistry, University of Szeged, Dóm Tér 8, H-6720, Szeged, Hungary

700    1_

$a Szczyrbová, Eva $u Department of Clinical and Molecular Pathology, Institute of Molecular and Translational Medicine, Palacký University Olomouc and University Hospital Olomouc, Hněvotínská 3, 77515, Olomouc, Czech Republic

700    1_

$a Eliáš, Martin $u Department of Clinical and Molecular Pathology, Institute of Molecular and Translational Medicine, Palacký University Olomouc and University Hospital Olomouc, Hněvotínská 3, 77515, Olomouc, Czech Republic

700    1_

$a Študent, Vladimír $u Department of Urology, Palacký University Olomouc and University Hospital Olomouc, I.P.Pavlova 6, 77900, Olomouc, Czech Republic

700    1_

$a Kurfürstová, Daniela $u Department of Clinical and Molecular Pathology, Institute of Molecular and Translational Medicine, Palacký University Olomouc and University Hospital Olomouc, Hněvotínská 3, 77515, Olomouc, Czech Republic

700    1_

$a Kovalová, Markéta $u Department of Experimental Biology, Faculty of Science, Palacký University Olomouc, Šlechtitelů 27, 78371, Olomouc, Czech Republic

700    1_

$a Mada, Lukáš $u Department of Experimental Biology, Faculty of Science, Palacký University Olomouc, Šlechtitelů 27, 78371, Olomouc, Czech Republic

700    1_

$a Bouchal, Jan $u Department of Clinical and Molecular Pathology, Institute of Molecular and Translational Medicine, Palacký University Olomouc and University Hospital Olomouc, Hněvotínská 3, 77515, Olomouc, Czech Republic

700    1_

$a Frank, Éva $u Department of Organic Chemistry, University of Szeged, Dóm Tér 8, H-6720, Szeged, Hungary. Electronic address: frank@chem.u-szeged.hu

700    1_

$a Jorda, Radek $u Department of Experimental Biology, Faculty of Science, Palacký University Olomouc, Šlechtitelů 27, 78371, Olomouc, Czech Republic. Electronic address: radek.jorda@upol.cz

773    0_

$w MED00001628 $t European journal of medicinal chemistry $x 1768-3254 $g Roč. 249, č. - (2023), s. 115086

856    41

$u https://pubmed.ncbi.nlm.nih.gov/36682291 $y Pubmed

910    __

$a ABA008 $b sig $c sign $y p $z 0

990    __

$a 20230418 $b ABA008

991    __

$a 20230425140925 $b ABA008

999    __

$a ok $b bmc $g 1924501 $s 1190084

BAS    __

$a 3

BAS    __

$a PreBMC-MEDLINE

BMC    __

$a 2023 $b 249 $c - $d 115086 $e 20230113 $i 1768-3254 $m European journal of medicinal chemistry $n Eur J Med Chem $x MED00001628

LZP    __

$a Pubmed-20230418