US 2OO)4O265809A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2004/0265809 A1 M0ras et al. (43) Pub. Date: Dec. 30, 2004 (54) POLYPEPTIDES DERIVED FROM RETINOIC Publication Classification ACID-RELATED ORPHAN RECEPTOR(ROIR) AND THEIR APPLICATIONS (51) Int. Cl.” ............................ C12Q 1/68; C07H 21/04; C07K 14/705 (76) Inventors: Dino Moras, Lampertheim (FR); (52) U.S. Cl. ......................... 435/6; 435/69.1; 435/320.1; Jean-Paul Renaud, Ostwald (FR); 435/325; 530/350; 536/23.5 Catherine Stehlin, Strasbourg (FR); Jean-Marie Strasbourg, Drusenheim (FR); Roland Schuele, Weisweil (DE); (57) ABSTRACT Eric Friedrich Greiner, Heidelberg (DE) The invention relates to polypeptides derived from the retinoic acid-related orphan receptor (ROR) in mammals, Correspondence Address: characterized in that they are delimited in their N-terminal ?RRET eXtremity by an amino-acid located between positions 1 to 2ND FLOOR 209, and in their C-terminal extremity by an amino-acid located between positions 450 to 452 of the rat RORß, o, or AIRLINGTON, VA 22202 (US) y, or by an amino-acid located at corresponding positions in nuclear receptor ROR of other Subtypes than C, ß and y, (21) Appl. No.: 10/477,116 and/or of the other mammals. The invention also relates to (22) PCT Filed: May 7, 2002 the use of these polypeptides, or of the molecular complexes 39 or the crystals containing them, for carrying out:—a process (86) PCT No.: PCT/EP02/05024 for the screening of a ROR-LBD ligand which is an agonist, or an antagonist of said receptor,—or a process for the (30) Foreign Application Priority Data analysis of the tridimensional structure of the complexes formed with said polypeptides, molecular complexes or May 7, 2001 (EP)........................................ O 1401175.3 crystals and a particular compound. 20 211 221 hRORY . - - - - - a * : K > T PEAPYASILT EIEHILVQSVC KSYRETC.R. RORY . • • • • • • • • • • APEVPYASILT DIEYLVQNVC KSEFRETCS:R hRORO. • • • - - - - - » GETSPTVSMA EILEHLAQNIS KSHLETCSÈL, TRORO. - - - - - - - - - - - - - - - gETSPTrvSMA ELEHLAQNIS KSHLETogg|L hRORß . • • • • • *GQLAPGIITMT EIDRIAQNII KSHLETCIJFET rRORß . GQLAPGITIMS EIDRIAQINII KSHELETCET N H1 hRARY . • • • • • • • • • • . DSYELSP QLEELITKVS KAHQETFPSL 204 ' 23 24. 25 * 261 271 W hRoRy LEDELRQRSN IFFSREEWTGY QRKSMWEMÄE I GA S.ŠAI QYV VEFAKRIL mRoRY LEDELRQRTIN LFSREEVTSY QRKSMWEM hRORO REENOOITWQ TEFLQE????? QN KQREVMS, mRORo REEQQITWQ TIFLQE????? QNKQREVM I QYV VEFAKIRI hRORß MEEËHQILAWQ THTYEEIKAY QSKSREA I QYV VEFAKIRI rRORß MEEGEHOLAWQ THTYEEIKAY QS O SI QYV VEFAKIRI 2 H2 * 3 K » hRARY . COLGK. YTTNSSADH RVQLD EGLWID KSEATS. IKIVEFAKR 2. O 9 28 22. 8 38 24 B 2 8 1 29 i 3 0) 31 321 hRORy SGFMELCOND CIVILLIKAGA | A YNADNRIFjºš - GKYGGME. mRORy SGFMELCQND QIILLTAGA. EVVY YNANINH GK YGOWE baRORO. EDGFMELCCND QVLLIKAGS V FDSQ KIYASP mRORO. DGFMELCONID QIVILLKAGS FDSQ GKYK ASP hRORß TOGFMELONO QILILLKSGC FNP GKNYGOM rRORß TGFMELCONID OILLLKSGCPw EV? t A FNP } \ , º?S Q hRARY PGFTGLSAD, QITILLKAAG4 DIKSLESET)H YTPEQDTM)êPGLTL?RTg. M 58 W 268 278 , '. 28 298 3 4 0 350 ' , 3 6 0 370 SSÈDFSHSIL SALHIFSIEDEI ALYTALVIN AHRPGILQEKR Ej , . SSDFSHFIL SALCIFSIEDEI ALYTALVILIN ANRPGILQEKR FGKS CSMHLITEIDEI AILFSAFVILMS ADRSWT QEKV F’GKSIL CSMHILTEDEI ALFSAFVLMS ADRSWILQEKV FAKNIL, CSILQILTE EEI ALFSSAVILIS PIDRAWLIEPR M SLOLIEEE ALFSSAVLISH8 PDRAWILLEPR.- hRARY MHNAGEGPILI3 C 8 DLVFAFAGQL.38 LP EMD?TET GILLSAICES3 3 8 GDRMDLEEPE3 4 8 Patent Application Publication Dec. 30, 2004 Sheet 1 of 10 US 2004/0265809 A1 Patent Application Publication Dec. 30, 2004 Sheet 2 of 10 US 2004/0265809 A1 GGGCAGTTAGCTCCCGGGATAACAATGTCTGAGATCGATCGAATTG CACAGAACACCIAT" - - - - - - - - - -- - - - - - - - - - -- - - - - - - - - - -- - - - - - - - - - -- - - - - - - - - - -- - - - - - - - - - + 6O CCCGGTCAATCGAGGGCCCTATTGTTACAGACTCTAGCTAGCTTAACGTGTCTTGTAGTAA G Q L A P G I T M S E I D R I A Q N I I – AAGTCCCAT"rTrGGAGACGTGCCAGTACACAATGGAAGAGCTCCATCAGCTGGCATGGCAG 65 - - - - - - - - - -+ - - - - - - - - --+- - - - - - - - - - v- - - - - s* - * - ~~ -- - - - - - - ~- - - -- - - - - - - - - - -- 2 0 TTCAAGGGTTAAACCTCTGCACGGTCAATGTGTTACCTTCTTCGAGGTTAGTCGACCGTTACCGTC K S H L E T C Q Y T M E E IL HI Q - L A W Q -- AccCACACCTATGAGGAAATAAAGGCATATCAAAGCAAGTCCAGGGAGGCTCTGTGGCAG 12 li -- - - - - - - - - - -- - - - - - - - - - --- - - - - - - - - - - - - - - - - - - - -- - - - - - - - - - -- 8 0 TGGGTGTGGATACTCCTTTATTTCCGTATAGTTTCGTTCAGGTCCCTCCGAGACACCGTC T H T Y E E I K A Y Q s i K s R E A L W Q – CAGTGTGCCATCCAGATCACCCACGCCATCCAATATGTGGTGGAGTTCGCCAAAGCGGATA L81 - - - - - - - - - -- - - - - - - - - - -- - - - - a - - - - - - - - - - - - - - -- - - - - - - - - - -- ~- - - - ~- - - - - + 24 0 GTCAACACGGTTAGGTCAGTGCGGTGCGGTTAGGTTTATACACCACCTTCAAGCGGTTTCGCCTAT Q C A I Q I T R A I Q Y V V E F A K R I ACAGGCTTCATGGAGCTGTGTCAGAACGATCAGATCTTACTTCTGAAGTCAGGTTGCTTG 241 - - - - - - - - - -+ - - - - - - - - - - -- - - - - ~ is - - - as - - - - s - - - - -- su as as - as ses as - ur -- - - - - - - - - - -- 3 0 0 TGTCCGAAGTTACCTCGACACAGTCTTGCTAGTCTAGAJATGAAGAACTTCAGTCCAACGAAC T G F M E IL ?. Q N - ID Q I L L L K S G C L GAAGTGGTTTTAGTGAGAATGTGCCGTGCCTTCAACCCATTAAACAACACTGTTCTGTTT 30 - - - - - - - - - -- - - - - - - - - - -- - - - - - - - - - -- - - - - - - - - - -+ - - - - - - - - - -- - - - - - - - - - -- 3 60 CTTCACCIAIAAJATCACTTCTTACCACGGCACGGCAAGTTGGGTTAATTGTTGGTGACAAGACAAA E v v L v R M C R A F N P L N N T v L F GAAGGAAAATATGGAGGAATGCAAATGTTCAAAGCCTTAGGTTCTGATGACCTAGTGAAT B 6 - - - - - - - - - -- - - - - - - - - - -- - - - - - - - - - -- - - - - - - - - - -- ~- - - ~- - - - - - -- - - - - - - - - - - -- 420 CTTCC'TTT'ATTACCTCCITTACGTTTTTACAAGTTTTCGGAATCCAAGACTACTGGATCACTTA E G K Y G G M o M - F : K A L. G. » S D D L v N GAAGCATTTGACTITTGCGAAGAATCTGTGTTCCTTGCAGCTGACCGAGGAAGAGATTGCT. 421 - - - - - - - - - -- - - - - - - - - - -- - - - ~- - - - - - -- - - - - -s es a- - - -- - - - - - - - - ---- - - - - -- - - - + 480 CTTCGTAAACTGAAACGCTTCTTAGACACAAGGAACGTCGACTGGCTCCTTCTCTAACGA E A F D F A K N L C S IL Q IL T E E E I A - 4, 8 CTGTTCTCCTCTGCTGTTCTG?TATCCCCAGACCGAGCCTGGCTGTTAGAACCAAGAAAA- - - - - - - - - -- - - - - ~- - - a- ur |- s- i- s- is hab - as - a- -- - - - - - - - - - -- - - - - - ~- - - - +- - - - - - - - - - + 54 0 GACAAGAGGAGGACGACAAGACTATTAGGGGTTCTTGGCTTCGGACCGACAATCTTGGTTTTCTTTT IL F S S A V L. I S P D R A W Lu L E P R K - FIGURE3: 2 Patent Application Publication Dec. 30, 2004 Sheet 3 of 10 US 2004/0265809 A1 GTCCAAGAAGCTTCAAGGAAAAAATTATTTTTTGCACTTCAACAATGTGATTTCAGAAGAATCAC 541 - - - - - - - - - -- - - - - - - - - - +- - - - - - - - - - -- - - - a- - - - ~ ~ -- - ~- - - - - - - - - - - - - - - - - - -- 6 O 0 CAGGTCTTCGAAGTTCCTTTTTTAAVATAAAACGTGAAGTTG”TACACTAAGTCTTCTTAGTG v Q K L Q E K I Y F A IL Q H v I Q K N H - CTGGATGATGAGACCCTGGCAAAGTTAATAGCCAAGATACCAACTATCACGGCAGTCTGC 6 (O) - - - - - - - - - -- - - - - - - - - - -- - - - ~- - - ou - -as -- ~- - - - - - - - - +- - - - --- - - - - -- - - - - - - - - - -- 6 6 0 GACCTACTACTCTGGGACCGTTTTCAATTATCGGTTCTATGGTTGATAGTGCCGTCAGACG AACTTGCATGGGGAGAAGCTACAGGTATTTAAGCAGTCTCATCCAGACATAGTGAATACA 661 - - - - - - - - -+ - - - - - - - - - H - - - - - -a - as am - - - - - - - - - - -- - - - - - - - - - -- - - - - ~- - - - - -- 72 O TTGAACGTACCCCTCTTCGATGTCCATAAATTCGTCAGAGTAGGTCTGTATCACTITATGT N L H G E K L Q v F. K Q S H P D II v N T – CTGTTTTCCTCCATTGTACAAGGAGCTTCTTTAATCCTGACTGTGCTGCGGTCTGCAAATGA 72 L - - - - - - - - - -- - - - - - - - - - -- ~ ~ - - - - - - - -- - - ~- - - - - - - -- - - - - - - ~- - - -- - - - - - - - - - -- 7 8 O GACAAAGGAGGAACAATGTTCCTTCGAAGAAJATTAGGACTGACACGACGCCAGACGTTACT IL F P P IL Y K E IL F N P D C A A V C K * FIGURE 2 (suite 1) Patent Application Publication Dec. 30, 2004 Sheet 4 of 10 US 2004/0265809 A1 2 0 211 221 hRORY . - • • • • • • • • • • TIPEAPYASILT EIEHILVQSVC KSYRETC.R. InRORY . • • • • • • • • • • APEVPYASILT DIEYILVQNVC KSEFRETCOR hRORO. - - - - - • • • * * GETSPTVSMA EILEHLAQNIS KSHLETCSFL mRORO. • • • • • • • • • GETSPTVSMA EILEHLAQNIS KSHLETCgšgL hRORß . • • • • • “GOLAPGITMT EIDRIAQNII KSHLETCSET rRORß . » GQLAPGITIMS EIDRIAONII KSHLETCST N H hRARY . • • • • • • • • • . DSYELSP QLEELITKVS KAHQETFPSL 204 24 25 * 261 271 FIS-re W hRARy . CQILGK. YTTNSSADH RVQLDEGLWD KESEATSI IKIVEFAKR 2 O 9 218 22. 8 ž38 248 281 29 . K hRORy SGFMELCQND QIVILLIKAGAY hRORß TGFMELCOND QILILLKSGC? EV? rRORß TGFMELCONID OILLLKSGCR EVSEV?c * 2 * 4) * * H4 H S ) š> hRARYA PGFTGLSEAD,258 QITILLKAAG* 2 6 8 DIKALEBET)278 YTPEQDTM ". êP GLITLRTQ298 W » 340 » 350 " , 3 6 O 370 SS. DFSHSIL SALHIFSIEDEI ALYTALV?L'IN AHRPGILQEKR V i NEADFAKNIL, CSILOLITIEEEI ALESSAVILIS PDRAWILLEPR 6 7 H8 V hRARy MHNAGÊGPL DLVFAFAGOL LPLEMIDDTET I GILLSAICLIC GDRMDLEEPE 3. O 8 3 18 33 8 3 4 8 Figure 3 Patent Application Publication Dec. 30, 2004 Sheet 5 of 10 US 2004/0265809 A1 3 8 0 3 9 0 4 0 0 4 09 419 . hRORy KVEQILQYNILE LAFHHHOLCKT HR., QS. ILAK LPPK. GKILIRS ESSQIš mRORy RVEHILQYNILE LAFHHHILCKT HR. QG. LLAK LPPK. GKILIRS ÈÈS hRORo. KIEKLQQIKIQ ILALQHVILQKN HREDG. ILTK LICKVSTILRA RES mRORo. KIEKLQQIKIQ ILALQHVILQKN HIREDIG. ILTK LICKVSTILRA È hRORß KVQKLQEKIY FALQHVIQKN HILDDE. TLAK ILIAKIPTITA GAGNL rRORß KVQK LQEKIY FALQHVI QIKN HILDDE . GNIL H Oa H OD H1 hRARY KIVIDKILQEPILL EALRILYARRR RPSQP MFPR MLMIKIITTOLIR STKAERAI 358 3 68 378 3 88 398 429 439 449 459 hRORY IFQHILHPIV V QAAFPPLY mRORY IFQHILHPIVV QAAFPPILÄÄKE LFSTDVESPE GILSK. hRORo. AFKAIYPDIV RILHFPPLIKE ILFTSEFEPAM QIDG. mRORO. AFKAIYPDIV RILHFPPLYKE LIFTSEFEPAM ? QIDG. hRORß VFKQSHPEIV INTILFPPLIKE LFNPDCATAC K. rRORß VFKOSHIPDIV NTT)F PDCAAVIC K. HI " H2 hRARY TEKMEJIPG,-