Efficient Lead Finding, Activity Enhancement and Preliminary Selectivity Control of Nuclear Receptor Ligands Bearing a Phenanthridinone Skeleton.

Yuko Nishiyama,Minoru Ishikawa,Makoto Makishima,Shinya Fujii,Yuichi Hashimoto

doi:10.3390/ijms19072090

Abstract

Background: Nuclear receptors (NRs) are considered as potential drug targets because they control diverse biological functions. However, steroidal ligands for NRs have the potential to cross-react with other nuclear receptors, so development of non-steroidal NR ligands is desirable to obtain safer agents for clinical use. We anticipated that efficient lead finding and enhancement of activity toward nuclear receptors recognizing endogenous steroidal ligands might be achieved by exhaustive evaluation of a steroid surrogate library coupled with examination of structure-activity relationships (SAR). Method: We evaluated our library of RORs (retinoic acid receptor-related orphan receptors) inverse agonists and/or PR (progesterone receptor) antagonists based on the phenanthridinone skeleton for antagonistic activities toward liver X receptors (LXRs), androgen receptor (AR) and glucocorticoid receptor (GR) and examined their SAR. Results: Potent LXRβ, AR, and GR antagonists were identified. SAR studies led to a potent AR antagonist (IC50: 0.059 μM). Conclusions: Our approach proved effective for efficient lead finding, activity enhancement and preliminary control of selectivity over other receptors. The phenanthridinone skeleton appears to be a promising steroid surrogate.

Highlights

Nuclear receptors (NRs) are ligand-dependent transcription factors that regulate DNA transcription by binding small molecular agonists such as hormones
Sci. 2018, 19, 2090 transporter 4 (GLUT4), are involved in lipid metabolism, reverse cholesterol transport, and glucose transport, so liver X receptors (LXRs) agonists are likely to be of therapeutic value in the treatment of atherosclerosis, hyperlipidemia, and metabolic syndrome
They are involved in the upregulation of sterol regulatory element-binding protein-1c (SREBP-1c) and fatty acid synthase (FAS), so LXRs antagonists might have therapeutic value for hepatic steatosis [13]

Summary

Introduction

Nuclear receptors (NRs) are ligand-dependent transcription factors that regulate DNA transcription by binding small molecular agonists such as hormones. Forty-eight structurally conserved kinds of NRs have been identified, and many of them recognize endogenous steroidal ligands (Figure 1). Because NRs control diverse biological functions including reproduction, differentiation, homeostasis, and the immune system, they were the targets of approximately 5% of FDA-approved drugs in 2011 [1]. A steroidal ligand, and its metabolites, brings with it the potential to cross-react with other nuclear receptors, which can result in unwanted side effects, potentially limiting the clinical utility of these agents. Surrogates for the steroid skeleton are required to develop selective non-steroidal NR ligands for clinical use [2]. Tcohmerpefoourne,dschoebmtaicinael dmboadsiefdicaotnionthiosfappoplryopahcahrmaraecloikloeglyicatol lpeoasdsecsosmppoolyupnhdasr, maaimcoeldogaictaol pchtiamriazcitnegr. sTehleecrteifvoirtye,fcohr eampiacratlicmuloadr itfiacrgaetito, nis oref qpuoirlyedp.hSairnmceactohleofgoilcdalstlreuacdtucroems opfoNunRdlsi,gaanimd‐ebdinadtinogpdtiommizaiinngs (sLeBleDctsi)viatryefsoirmailpaar,rtwiceulhaarvtearugseetd, isthriesqsutirraetdeg. ySitnocecrtehaetefollidgasntrduscbtueraersinogf aNdRiplihgeannydl-mbientdhiannge dskoemleationns (foLrBsDesv)eararel NsimRsi,lainr,cwluedhinagvefaurnseedsotihdisXsrtreacteepgtyorto[3c],reliavteerliXgarnecdespbtoerar(iLnXgRa) d[4ip],hveintaymlminetDharneceespkteolret[o5]n, afonrdsreovgeernalreNceRpst,oirn(cAluRd)in[6g],faanrndeessotirdogXernerceecpetpotro[r3[]7, ]l.iver X receptor (LXR) [4], vitamin D receptor [5], androWgeenhraevceepptroerv(iAouRs)l[y6]d, eavnedloepsterdogLeXnRresceapnttoagr o[7n]i.st 1 [8], retinoic acid receptor‐related orphan recepWtoersha(vReOpRrse)viionuvselrysedeavgeolonpisetd 2LX[9R]s, apnrtoaggeosnteisrton1e[8r]e, creeptitnooric(PaRci)d arnectaegpotonri-srtela3te[d10o],rpahnadn prehcaerpmtoarcso(lRoOgiRcsa)lincvhearpseeraognoensist42 [a9n],dpro5gefsotreroNnieemrecaenpnt‐oPri(cPkR)dainsetaagsoenitsytp3e[10C],1an(dNpPhCa1r)ma[1co1l]ogaincadl Nchiaepmearnonn‐ePsi4ckandty5peforCN1‐ielimkean1n-P(iNckPCd1isLe1a)se(tFyipgeurCe1 (1N) P[C112)].[11T]haensed Ncoiemmpaonunn-Pdisckatlyl pecoCn1ta-liinke a1 (pNhPenCa1nLt1h)r(iFdiignu‐6re(51H) [)1‐o2n].eTshkeesleectoomn paosuandcyscallilzceodntcaairnbaa‐pahneanloangthorfidthine-6s(k5eHle)t-oonneosfkeLlXetRosn pasana caygcolinziesdt Tca0r9b0a1-3a1n7al(o6g).oBfetchaeusskeeleentdonogoefnLoXuRs slipgaanndasgofonristthTe09a0b1o3v1e7 p(6r)o.teBiencsau(LsXe Rens,dRogOeRnso,uPsRli,gNanPdLsCf1oratnhde NabPoCve1Lp1ro) taelilnhsa(LvXe Rass,tReOroRids, sPkRe,leNtoPnL,Cw1 eanhdypNoPtChe1sLi1z)eadllthhaavt eifatshteerpohidenskaenltehtorind,iwn‐e6(h5yHp)o‐tohneesiszceadfftohladt aifctthseapsheanasntethroriiddins-u6r(5roHg)a-oten,e pschaefnfoalndtahcrtisdainsoanseterdoeirdivsautrirvoegsatwe,opuhlednaanlsthoribdiinndonteodeortihveartivNesRws otuhladt raelscoogbninizdeteonodtohgeernNoRuss sthteartoriedcaolglingiaznedesn.dTohgeenapopursosatcehrodideascl rliibgeadndhse.rTehiseeaxpppercotaedchtodebsecruisbeefdulhienrethies dexepveeclotepdmteonbteouf sneofvuellinsttehroeiddesvuerlroopgmateenst, osuf cnhovaesltshteerpohidensaunrrthogriadtienso, nsuecshkealsetthone,pehneanbalnintghreidffiincioennet lsekaedletfoinnd, ienngaboflinliggaenffidcsiefnort lteaardgefit npdriontgeinosf ltihgaatnrdescofogrnitzaergeent dporgoetenionussthsatetrroeidcoagl nliigzaenednsd, oagnednothues gsteenreoridatailolnigoafndmso, arendsetlheectgiveneelriagtainondsofbmy ofurertsheelercmtivoelelcigualanrdsmboydfiufirctahteiornms oolef cnuelawr manoddicfiocnavtieonnisenotf snceawffoalndds c(oenasvyentoiensyt nscthafefsoilsd, sea(esaysytotoinstyrondthuecseiss,uebassytittuotiennttr(osd) uacteansuybpstoitsuittieonnt,(se)tca.t),acnoymppoasirteiodnt,oettch.e), sctoemropiadresdkteoletthoens. teHroeirde,skweleetosnh.oHwerteh, awt easphpolwicathtiaotnapopflictahtiisonaopfptrhoiascahpptrooaocuhrtosomuarlsl mliabllralirbyraoryf pofhpenhaenntahnrtihdriindoinneo‐nbea-sbeadseRdORROsRins vinevrseersaegaognoisntisstasnadndPRPRanantatgaognoinsitsstsleldedtotosseevveerraallsseelleeccttiivvee LLXXRRββ,, AARR aanndd gglluuccooccoorrttiiccooiidd rreecceeppttoorr ((GGRR)) aannttaaggoonniissttss

Results and Discussion

T-47D Alkaline Phosphatase Assay

Reporter Gene Assay

PSA Level in LNCaP Cell Line

Conclusions