HUE030079T2 - Cisztein-manipulált antitestek és konjugátumok - Google Patents

Cisztein-manipulált antitestek és konjugátumok Download PDF

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HUE030079T2
HUE030079T2 HUE05811919A HUE05811919A HUE030079T2 HU E030079 T2 HUE030079 T2 HU E030079T2 HU E05811919 A HUE05811919 A HU E05811919A HU E05811919 A HUE05811919 A HU E05811919A HU E030079 T2 HUE030079 T2 HU E030079T2
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antibody
seq
frac acc
cysteine
receptor
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HUE05811919A
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Charles W Eigenbrot
Jagath Reddy Junutula
Henry Lowman
Helga E Raab
Richard Vandlen
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Genentech Inc
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Claims (6)

  1. CISZTEiN-MANl^ULÁLT ANTITESTEK ÉS KONJUGATONIOR Szabadalmi ifêny pontok
    1, Eljárás oiszieírwimnipuláli (aiszMinmnginaared} antitest előállítására» amely antitest tartalmai egy vagy több olyan szabad elszteín amlnoeavai amelynek a tícslreaktívstási értéke a 0,6 -1,0 tartományba esik. és a nehézfáncában van olyan szekvencia, amelyet a SEQ ID NO; 10,11, 13, 14, 31,34, 37, 38 és 42 közül választunk: NSLRCEDTAV (SEQ ID NO: 10) LVTVCSASTKGPS (SEQ ID NO: 11} LVTVSSCSTK.G PS (SEQ ID NO. 13} LVTYSSAC-TKGPS (SEQ ID NO: 14} HEDPEVKFNVWVDGCEV'HNAKTKPR (SEQ ID NO:31) KGFYPCDÍAVE (SEQ ID NO:37) PPYLDCDG5FF (SEQ ID Nö;38) GQaTLVTYSACSTKQPSVEPL (SEQ IP Nö:42) ahol a szabad cisztáin amlnosav a SEQ lö NO: 10, 11, 13, 14, 31, 34, 37. 38 és 42 szerinti szekvenciákban lévő cisztein; vagy a kénnyűlánoában van olyan szekvencia, amelyet a SEQ ID NO; 17, 19, 29, 21,22, 24,25 és 27 közül választünk: SLSASCGDRVT (SEQ ID NO;17) ElKRTCAAFSV (SEQ ID NO: 1S) TCAAPCYFIFRP (SEQ ID NO:20) FIFPPCOEQLK (SEQ ID NO:21) DEQLKCQTASV (SEQ ID NO;22) WKV&NQiÄqN- (SEQ ID Nö'24) ALö8CIQSQES¥ (SEQ ID NO:25) C3LSSPCTKSFN (SEQ ID NO:27) iicsl a szabasl olsztein amtnosav a SEQ ID NO: 17, 19, 20, 21., 22, 24, 25 is 27 szerinti szekvenciákban lévő cisztáin; ahol az eljárás sarán egy kiindulási („szülő”) antitest egy vagy több áminösiy-maradékát áeípitesítjök szabad cisztein aminosavmiaíadék{ök}kaí, abet a kiindulási antitest szelektíven képes kötődni egy antigénhez, és a oiszteim szerkesztett antitest szelektíven képes kötődni ugyanahhoz az antigénhez, mint a kiindulási antitest, és ahol a eíszteimszerkesztett antitest humán, kimére vagy humanizál antiest. t, Az 1. igénypont szerinti eljárás eiszteimrnanipyiált antitest előállítására, amely antitestet ofyen eprással állítunk élő, amelynek a során: II mutégenizáiUhk nukleinsav-szekveneiát, hogy kódolja a cisztein-szerkesztett antitestet; (ii) exprésszáitatjuk a císzteimszerkesztett áhtltestet; és (Ili) izoiáijuk és tisztítjuk a cisztein-szerkesztett antitestet.
  2. 3, A ti. Igénypont szerinti eljárás cisztein-manipuíáit sotiást előállítására, amelynek során továbbá: (i) reagáltatok a dsztein-szerkesztett antitestet tioíreaktiv affinításreagenssei, affinitásjelöft, cisztein-szerkesztett antitest előállításához; és (ii) megmérjük az atfinitásjeiölt, císzteín-szerkesztett antitest kötődését egy kikötő közeghez. 4 A 3. igénypont szerinti eljárás eiszteímmanipuláít antitest előállítására, ahol a tioiréaktlv affinításreagens tartalmaz biotin molekularészt és maíeímíd molekularészt, ä 3, igénypont szerinti eljárás olszfein-manlpy Iáit Ältest előállítására, ahol a kiköt# kőze$ ta?talmáz sztrepíavidint S< Az előző Igénypontok bármelyike szerinti eljárás oiszteínmianipulátí antitest etőálliásirá, ahol a kiindulási antitest fúziós fehérje, amely tartalmaz a következők közöl választott albuminkotő pepiidet (ABFj: üümHTúmBmiMwicí^múm^mw cseq id no· i>; (seq ip » 2), QRUEOlCtPRWGCLWEDDF (SEQ ID NO: 3), RLIEDICLPRVVGCLWEDD (SEQ IP NO: 41 és DICLPRWG.CLW (SEQ ID NO: 5).
  3. 7, Az előzd Igénypontok bármelyike szenniteljárás cisztein-manipeláft antitest előállítására, ahol a kiindulási antitest monoklonáiis antitest, bispeciíikus antitest vagy antifestfragmens. 8, A 7. igénypont szerinti eljárás dsztesn-manipulált antitest előállítására., ahol a kiindulási antitestet huy.Ab4D5~8 (trasztuzumab), an?bEphB2R antitest és anti-MUCI # antitest közül választjuk, B. Az előző igénypontok bármelyike szerinti eljárás clsztein-manipuláit antitest előállítására, amely tartalmaz olyan aminosav-szekvenciák amelyet a SEQ SD NO.6 (azaz 6. sz, szekvenoia): EyQLVESGeOLVQPCKkSLMACAASGFNIKDTYmWVROAPOKÔLlWmœrrNlIYllY A0SVKCjRFnSAD7SKNTAYLQSINSLRCEDTÀVYYCSKW(K.sDôFYAMDYWôCpTL¥ï¥SS ASTKGPSVFPl,APSSKSTSCrrTAALCk:EVKDYFPEPWYSÄlGAi,TSGVlfTFPA\^QSS Gl.,YSLSSV¥'rV"PSSSmTQTYÏCm/NHiKPSHTKVDKK¥lPKSCI>KTftTCPK';PAPELLGCs FS¥FlFPFKmtYttM!SRTPHWCV¥¥ri¥SMEDPE¥lMWYVDGVEVHNAKTKPMQyM ^TYRWm.tVLHQDWLNGKEYKCKVSNKAlLPAKEK'nSKAKOQPiyBKîVYTLPPSREE MTKNQVSLTCLVKÔFYPSDWVEWESNGQPBNiîYKTOPVjUgSPOSFEtYSia^mW QQGWFSCSVMMEALMNMYTQKSLSLSPGj, vágy EËQ ID NO:7; WQLVg^SSÍÖLVQP(KíStlU^CAA^FNIK»TV1HWVR0AK#KÖLlW¥ARI'yFrHaY,3Íty AÖSVKGRFUSADTSKNTAYLQMNSLRAJEtyrÄVYYCSRWGöDGFV'AMDYWCIQÖTLWVSS GSTKGMVFFWSSKSÏ$OOTAALGCLVK.DYFPEFY'rVSWNSGAL«WTFRAVLQSS CR^SLSSVVTVFSSSÎ^T^YlCKVNHKPSm’KVDKJK.VÊFKSCDKYHYÊ-RPC^EiXOS PSYFLFFFKFK-DTlMiSRTPKVTCrVWBYSHHOMYKFHWYYBOVEYllMAKTRPREEQYN STYRVVS\aiYTHQDWL^GKEYKCKYSR^RAy>AMEKm3KLAKÖQPliWTTLPRSRRH MTKHQVSI¾E^tóFYPSDïAVBWES^Î<K?FliÄ¥RYYIPÄPSÛCïSFFL·YPÎI^'Y©KSRW «^^NVFSCi^MHEAEH'MHYYQKStJlSPClKi YRgy ÊBêM MQ:B: DÎQMTQSPS$LSASVaDRVTÏTCRASQDVNFAVAWYQQKPGKAPKUJY$A$FLYSGV?*S RFSGSRSGTDFTLTÏSSLQFEDFATYYCQQKYITPPTFGOGI'KVEÎKRÏCAAPSVYIR^P SDFQLKSGlV\SV'VCLLNKFY?R£AKYOWRVDNALQSONSQÈS\TB;>DSKDSTYxRL.SSTI.:t LSKA DYEKHKVYACF VTHQGiJSSPVTKSFMRGEC, vagy SEQ ID NO:28: EVQLVESGCjGLVQPGGSLRLSCAASGYT.FTSYWMMWVRQAFCiKGLEWVGFmPSTGY®¥ NQKFKDRFTISADI'SKNTAYLQMGSI.RAEDTAVGTCTRR.PKÏPRHAN'YFWüiXTrLVTVSS CSTKG'FSYFFLAFSSKSISGCîTAALGCLVKÔYFFEFVWSWNSGALTSGVHTFPAVLQSS Gl,YSLSSWTVFESSLGTQTYlQYVNHKPSNlKYT.>KK¥EPKä€i>K'föT€PP€PAPELLöö PSVFLFFPKFKDîLMIS&amp;'rFBVl'CWVDVSHEDPEVKJFNWYVÏXjVEVMNAKTKPREEQYH SI’YRVVSVLTVl.HGDWLNGKEYKCKVSMIG^I.PAPIERYISKAKCîQPRFPQYYTLFFSREE MTR^QVStTCLVKGFYFSDIAVEWES^GQFENNY'OTPPVLDSiXiSFFLYSKETVDKSRW OíXsNVFSCSYMHEALHHHYTQRSESLSFölC, és SEQ m NO: 39: ÔVQfc#ESGPGLVM>SQSLSLTCTYrGYSITNDYA^m'IRGFFGNKL.EWMGYrNYSGYTTY NFSLRSS.ISITRDTSKNQFF1.HLNSVTTBDTATYYCARWDGGLWWGQGTLWSACSTK <3FSVFPliAPSSKSTSGGTAALGCLYKDYFPEPV,FYSWHSÔAL'fS0VHTFPAVK3®i^jLYS LSSY'VYYFSSSLGTQTYÎCFTV'iNHKFSHTKVDKKVEPKSCDKTHTCPFCFAPELLGGFSVF LFPFKFKBTtMÏSRTFFA'T€YYYI>VSHEDPEVKFM\WTC¥EV,HNAKTKPREBQYNSTYR WSVLWLHQDWLNGKEYKCKVSNKALPAPIEKTISlCAKCiQPREPQVYTLPPSRjEIMTKH QVSlÂIA/KÔFYFSDIAVEWESNCK^FENNYKTtFFYLDSDGSFFÉYSKlTVlDKSRWCJQGN WSCSYMHEALHNHYTQKSESLSPÖIC közül választunk.
  4. 19, Az 1-7. igénypontok bármelyiké szerinti eljárás ciszleinmianipüláíi antitest előállítására, ahoi a kiindulási antitest IgA, IgD, IgE, IgG és IgM közül választói Intakt antitest. ^ fénypont szerinti eljárás eísziein-manipuláit antitest előállítására,, ahol a következő alosztályok közül válaszíjuk: igGI, tgG2, lgG3 és !gG4. ^->ÿ y| : ^ ....... ^ . * vaSV 10-ii, igénypontok bármelyike szerinti e Ijá rás eíszfeí n -m a η I pu í á it antitest élöáíiíté$§ra ahol a císztain-manipulált antitest vagy a kiindulási é elitest képes kötődni egy vágy főbb alábbi f;1~36. sorszámul receptorhoz: (1) B&amp;fPR-jii (is típusú caont-moriogenetskus fehérje receptor); (2) £16 (I.AT1 SLC7A5); (¾ STBAP1 (a prosztata hat transzmembrán doménes epiteiiális antigénje): (4) Ö772P (CA125, MUC16); Cí) ypp (MPF. MSLN, SÍVÍR, megskariocita-potencírozé faktor, mezotekn); M Néplap flAPI-GB, N'FTÍÍb, SLG34A2, oldott anyag hordozó család 34 (nátrium-fo$zfát); 2. tag, íí-es típusú nátríumfüggö foszfáttranszporter-Sh); C7| Soma 5b (FU 10372, KIAA1445, Mm.42G15, SEMA58, SEMAG, Szemaforin 5b Hlog, szama donién, hát irombospondin ismétlődő régió (1~ típusú és 1 -eshoz hasonló típusú), transzmerohrán dómén (TM) és róvip oítoplazmás dómén, (szemaforin) 5B); P) PSCA Híg (27ÖööSöC12Rik, C530ööSO16Rik, Rí KEN cDMS 270D0iöGi2t RÍKEN cDNS 2709Ö6ÖC12 gén); (9) ET8R (B típusú endotei in-receptor); (10) MSG783 (RNF124: FLJ20315 hipotetikus fehérje); (11) STEAP2 (HGNC.J639, ÍPCAG, PCANAP1, BTAMP1« STEAP2, STRIP, prosztatarákhoz kapcsolódó gén 1, prosztatarákhoz kapcsolódó fehérje 1, a prosztata számú hát transzmemhrán domines epiteiiális ihtigénle, heltrenszmémbrln domines proszta tafehérje); (12) TpM4 (BR22450, FLJ20>041, ÎRPM4, ÏRFM48, tranziens reoeptorpotencíál kationcsatorna, M aicsaíád, 4. tag): (13) ORíPTO: (CR, CR1, CRGF, CRiPTÖ, TDGF1, teratökarcinóma-eredetű növekedési faktor);: (14) 0P21 (CR2 (2-es komplementreceptor) vagy C3DR (C3d/Epsteín- Barr virus reeeptor) vagy Hs.73792); :(1-5) CD79h (CD79B, CD79jl. H3b (immunglobuíin^asszociélt bétá), 829):: (if) FcRH2 (IF0P4, IRTÄ4, SEAR 1.A (SH2~doment tartalmazó foszfafázhotgony fehérje, la típus), SPAP1B, SPAF1C}; (17) HËR2; (18) MCA; (19) MDF; (20) IL20Rcr; (21) Brevlkán; (22) EphB2R; (23) ÂSLG659; (24) PSCA; (25) GEÖA; (28) BAFF-R (Bmejt aktíváié faktor receptor, 3-as Bty8 receptor, BR3; (27) CD22 (B'-sejt receptor CD22-B izoterma); (28) CD78a (CD79A, CP79o, imrnungiöhuiimasazoüiált alfa, agy S-seji specifikus fehérje, amely kovalensen kölcsénhat az tg-bétával (OD79B) és a felszínen IgM molekulákkal komplexet képezve transzdukál egy, a B~ sejtek differenciálódásában szerepet játszó szignált); (29) CXCR5 (Ims Burkit-téle limféma-receptor, egy G-fehérjéhez kapcsolt receptor, amelyet a CXCL13 nevű kemokín aktivál, közreműködik a llmfoclták migrációjában és a humorális védelemben, szerepet játszik a HIV-2 fertőzésben és esetlég az AIDS, a íímfőma, a mleléma és a leukémia kialakulásában Is); (38) HLA-DOB (peptideket kötő és a GD4* T-llmfociták számára azokat preze?itáló II, osztályú MHO molekula (la antigén) béta alegysége); (31) P2X5 (ligandum-kapuzott ioncsatoma P2X purinreoeptor 5, egy ioncsatoma, amelyet extraoelluiárís ATF kapuz. és amely részt vehet szinoptikus transzmisszióban és neurogeoezlshen, hiánya hozzájárulhat az idiopátlás defrúzor-ínstabifitás patöfiziolögiájához)· (32) CD72 (CD72-es B-sejt differenciálódási antigén, tyh~2); (33) LY84 (04-es íimfbcitaantigén (RF1Ö5), Fos típusú membránfehérje a leucin ban gazdag ismétlődő régié (ORR) családból, szabályozza a B-sejtek aktiváeióját és apoptőzisát, funkciókiesése szisztémás lupus erythematosus-han szenvedő betegekben növekvő bétegségaktiuitáshoz kapcsolódik}; (34) FcRH 1 (Fc reoepferszerü fehérje 1, az immunglobulin Fc-domén egy feltételezett receptora, amely €2 típusú lg~szerú és ITÄfe doméneket tartalmai, szerépét jifszbát a Bdimfeeifák dtlfemooiiiédásáöan): (35) I1ÎA2 (Immunglobulin szupercsalád receptor, transzíokáció asszociált 2*es, egy féltéfeiéiett immunreoéptor, amelynek szerepe lehat a B-sejfek fejlődésében és limfőmagenezssben; egyes 8-sejfes mallgpanclákbái a gép trarifzlökáeió révén deregulálódik); és (SS) f®Í2 (feltételezett transzmembrán profeoglikán, kapcsolatban áll a növekedési faktorok EGF/heregulln családjával is a folliszfatinnai). 13« Az előző Igénypontok bifmelylke szerinti apris cJszteln-manlpyiált antitest előállítására, ahol az antitest kovalensen van kapcsolva kifogás! jelöléshez, detektálási jelöléshez vagy szilárd hordozóhoz, 14, A 13, igénypont szerint eprás ciszfein-manipuiált antitest előállítására, ahol az antitest kovalensen van kapcsolva fluoreszcens festékéé detektálási jelöléshez, amelyet fiuoreszceindlpusú, rodaniinóipusú, danzil, Nsszamin, egy cianln, egy íikoeriírín, Texas-vörös és ezek analógja közöl válesztuhk, fi» A 13, igénypont szerinti eljárás eisztein-manipylált antitest előállítására, ahol az antitest kovalensen van kapcsolva redíonyklid detektálási jelöléshez, amelyet 3H, 11C, i4C, 18P\ 3¥, 3S8, S4Cu, 63Ga, æY, δ8Το, ι;>?1!η, m\, 124l, 12Si, 131í, 133Xef 17-Lu, si 'At is S13BI közéi választunk, 13« A 13, igénypont szerinti eljárás císztein-manipulálf antitest előállítására, ahol az antitest kovaiénSéi van kapcsolva detektálási jelöléshez kefátorligandon keresztül, amelyet DOTA, OOTP, ©0TMA, »PA és fim közöl választunk. 1?, Antitest-hatóanyag konjugálum vegyüiet, amely tartalmaz cisztein-szerkeszett antitestet (Ab), amely tartalmaz olyan szabad ciszíein aminosavat, amelynek a tíoireikfivitási értéke a 0,8 - 1,0 tartományba esik; és a konjugátum tartalmaz hatóanyagmolekuiarészf (D), amelyet egy rnajtanzinold, egy aurisztatin, egy dolasztafin és egy kaíikeamsein közül válásztunk, ahol a ciszfeln-minlpuláif antitest egy vagy több szabad dszteín amlnosavön ker&amp;tóői, kapcsoló (linker) molekularészen (L) át kapcsolódik a D~hez; az I. képlet szerinti vegyöíst: Ab-{L-D)i;> î ahol p jelentése 1, 2, 3 vagy 4; és ahol a císztdnmisnipulátt antitest előállítása olyan eljárással történik, amelynek során a kiindulási antitest egy vagy több aniinösav-maradékát egy vagy több szabad essziein aminosevvaí helyettesítjük, ahol a kiindulási antitest: képes szelektíven kötődni egy antigénhez, és a dsztein-manipuláit antitest képes szelektíven kötődni ugyanahhoz az antigénhez, mint a kiindulási antitest, ahol az antitest·· hatóanyag konjugátuni vegyüiet tartalmaz: egy vagy több olyan szekvenciát a nehézláncban, amelyet a SEC? ID NO. 10, 11, 13,14, 31, 34, 37 és 38 közöl választunk: NSLRCEDTAV (8EQ ID HO: 10} LVTVCSASTKGPS (SEQ ID NO: 11) LVTVSSCSTKGPS {SEQ ID NO: 13) LYTVSSACÏ KG PS (SEQ 10 NO: 14) HEDPE¥KFNWWDGCEVNNAKTKPR (SEQ ID NO: 31} YKCKVCNKAl.P (SEO ID NO: 34} KGFYPCDtAVE ÍSEQ ID NO: 37) PPVLDCDGSFF (SEQ IP NO: 38} GQGTtVTVSACSTKGPSVFPL (SEQ ID NO: 42} ahöl a szabad dszfeln anilnosáv a SEQ 10 NO: 10, 11, 13, 14, 31, 34, 37, 38 és 42 szerinti szekvenciákban lévő eísztein; vagy egy vagy több olyan szekvenciát a könnyöláncban, amelyet a SEQ ID NO: 17, 19, 20, 21, 22, 24, 25 és 27 közöl választunk: SLSASCGDRVT (SEQ IP NO: 17) EIKRTCAAPSV (SEQ ID NŐ I9) TCAAPCVFIFPP (SEQ ID NÜ:29) FIFPPCDEQLK (SEQ ID NÖ:2í) DEQLKOGTASV (SEQ ID NO:22) WKVDNCLQSGN (SEQ ID N0:24) AL0SGCSGESV (SEQ IO NOî26) GtSSPOTKSFN (SEQ ID M0:2?) aho! a. szâbgd ciszteip aminosav a SEQ ID NO: 17, 19, 2Ô, 21, 22, 24,. 25 és 27 szerinti szekvenciákban lévő cisztáin, és ahol a cisztein-manipülllt antitest humán, ksmèra vagy humanizált antitest, 18, A 17. igénypont szerinti antitest-hatóanyag konjugátum vegyulet ahoi a olszteffvmanipulált antitest olyan eljárással karul elesilitásra, amelynek során: (a) egy kiindulási antitest egy vagy több aminosav-maradékái cisztelnnef helyettesítjük; és (b) meghatározzuk a oisztein-manipuiáit antitest tiolreaktivitását oly módon, hogy a cisztein-manípulált antitestet tiolreagenssel reagáltatjuk; ahol a elszieirvmanipulált antitest reaktívabb a thireaganssel, mint a kiindulási antitest, 19. A 17, igénypont szerinti antitesEbatóanyag konjugátom vegyidet, amely tartalmaz továbbá albuminkötö pepiid (ÄBP) szekvenciát, amelyet a következők közül választunk: SEQ ID NO: 1, SEQ IP NO: 2S SEQ ID NO: 3, SEQ ID NO: 4 és SEQ IP NO: 5. 29, A 17. igénypont szerinti antitest-hatóanyag konjugátum vegyulet, aboi a cisztein-manipulált antitest képes kötődni olyan ErhB receptorhoz, amelyet a kővetkezők közöl választunk: EGFR, HER2, HER3 és HER4. 21. A 17. igénypont szerinti arkitest-hatéanyag konjugátum vágyóiét, ahol a eiszteinmiardpuláii antitest vagy a kiindulási antitest képes kötődni egy vagy több alábbi (1-36, sorszámú) receptorhoz: (1) ByPRIB (IB típusú esont-morfogenetikus fehérje receptor); (2) E16 (LATI, 8LC7A6); fi) 8TEAP1 (a prosztata hat transzmembrán deménes epíteliálss antigénje); (4) Ö772P (CA126, MUCI6); (5) MPF (MPF, Μ Sill, S MR, megakariodta-poteneirozo faktor, mezofelin}; (6) NapiSb (NAPI-'38; MPTtlb, SLC34A2, oldott anyag hordozó család 34 (nátrium-foszfát), 2, tag, INs típusú nátriumfüggő foszfáí£fanszpo?tar~3h); (?) Séma 5b (FIJI03?l, KIAA1445, Mm.42015; SEMA58, SEMAG, Szemaforin 5b Hlog, szama dómén, hét trombospondin ismétlődő régió fies típusú és az 1-eshez hasonló típusú), transzmembrén dómén (TM) és rövid cítoplazmás dómén, (szemaforin) 5B); (8) PSGA híg (2790050012 Rík, C53ÖÖ08Ö18Rik, RIKEN cDNS 2706059012, RIKEN pPNS 27ÖÖÖS0G12 gén); (9) ETBR (B típusú endoieltn-receptor); (11) STEAP2 {HGNC_8639, IPGA-1, PCANAP1, STAMP1, STÉAP2, ΒΎΜΡ, prosztatarákhoz kapcsolódé gén 1, prosztatarákhoz kapcsolódó fehérje 1, a prosztata 2. számú hat transzmembrán dornénes epsíeliálís antigénje, hat transzmembrán doménes prosztatafehérje); (12) Trpy4 (8R22450, FLJ20Ö41 TRPM4, TRPM4B, tranziens fecepíorpotencíál kationcsatorna, M aicsalád, 4, tag); (13) CRÍPTO (GR, CRI, CRGF, CRÍPTO, TDGF1. teratekarcinóma-eredetű növekedési faktor); (14) CD21 (0R2 (2-es komplernenirecepíor) vagy C3DR {C3d/Epstein-Barr vírus receptor) vagy Hs.73792); (15) CD79b (CD79B, CD79p, IGb (Immunglcbuiín-asszöcíáít béta), 1329); (16) FcRH2 (ÍFGP4, IRTA4, SPAP1A <SH2~doméni tartalmazó foszfofázhorgony fehérje, la típus), SPAF18, SPÄP1G); (1?}BER2; (18) NCA; (20) IL2öRo; (21) Srevikán; (23) ASLG659; (24) P8CA; (25) GEDA, (26) BAFF-R (B-seji aktíváié faktor receptor, 3-as BLyS receptor, BR3; (27) CD22 (B~sejt receptor CD22-B izoforma); (28) CD? 9a (C079A, CD?9ö, immungIohu I irvasszoei âit alfa, agy B-sejt specifikus fehérje, amely kovalensen köfesdnhat az fg~béfával (CP79B) és a felszínen \gM molekulákká) komplexet képezve transzdukái egy, a B-sejtek differenciálódásában szerepet játszó szignált); (29) CXCRS (1-es Burkitt-féié limfomaaeceptor. egy G-fehérjéhez kapcsok receptor, amelyet a CXOL13 nevű kemokin aktivál, közreműködik a limfocitak migrációjában és a Immorális védelemben, szerepet játszik a :HJV~2 fertőzésben és esetleg az AIDS, a iimfónia, a mielőma és a leukémia kialakulásában is); (30) HLA-DOB (pepfideket kötő és a 004+ T-Iimfoclták számára azokat prezentáló Si osztályú SVfHC molekula (la antigén) béta alegysége); (31) P2X5 (Itgandumkkapuzott: ionesatcma R2X purinrecepfor 5, egy ioncsatorna, amelyet extraceiiuSáns ATP kapuz, és amely részt vehet szinaptikus transzmisszióban és a hiánya hozzájárulhat az idiopátiás öefrúzoNnstabilSiás patofizioSégiájához): (32) CD72 (CD72“es B~sají differenciálódási antigén, Lyb-2); (33) LY84 (64-es iimfocifaantigén (RSP1Ö5), i-es típusú membránfehérje a ieucinban gazdag ismétlődő régió (LRR) családból, szabályozza a 6~ sejtek aktiváoióját és apoptózisát, funkciókiesése szisztémás lupus erythematosus-ban szenvedd betegekben növekvő betegaégaktivifáshoz kapcsolódik); (34) FcRHi (Fc receptorszerű febérjé 1, az immunglobulin Fc~domeo egy feltételezett receptora, amely 02 típusú Ig-szeru és ITAM doméneket tartalmaz, szerepet játszhat a Bdimfociták differenciálódásában); (35) SRTA2 (immunglobulin szuperesáiád receptor, transzlokáoio asszociált 2-es, egy feltételezett immuoreceptor, amelynek szerepe lehet a B-sejtek fejlődésében és iimfőmagenezlshen; egyes B--seJtes mailgnanofákban a gén irsnsziokáeiö révén deregulálódlk): és (36) TÊNB2 (feltételezett tranazniembrán proteoglikán, kapcsolatban áll a növekedési faktorok EtiF/hereguiin családjával és a fejlisztatlnnal). 22* A. t?< igénypont eaerlntl abtltésf-hatóanyag konjugatym vegyület, adói p értéke 1 vagy 2, 23. A 17. igénypont szerinti antitest-hatóanyag konjugátum vegyülei ahol L képlete a következő; -ArWw-Yy» ahol; A Jelentése stretcher (nyújtó- vagy feszítő} egység, amely kovalensen kapcsolódik a cisztein-manipulált antitest (Ab) egy clsztein tioíjához; a értéke Ö vagy 1 : mindegyik VY jelentése egymástői függetlenül aminosav-egység, w Jelentése egész szám, értéke 0 -12; Y jelentést távtartó (spacer) egység, amely kovalensen kapcsolódik a hatóanyagmolekularészhez; és y étéke 0,1 vagy 2, 24. A 23. igénypont szerinti antitest-hatóanyag konjugátum vegyüfei amelynek képlete a következő;
    ahol a RAB jeientése para-arninohenziikarbamoii; R1? jelentése dlvaiens csoport,, amelyét (CH?)?, Ca-Og karhoolkl, O-ÇCM?)?, arllétr, (CHgjranlén, -arilén-(CH2)r*5 (O.Hg)r(C:3“C§· karbocikii!}, {CrCg karboPikJi!HOH2}f> Cg-Og heteroeikiil, (CH2)r(CrCa hetaoeHtfi). "(Ca-Cg hatar00iki)-{CH2)ri -t(.CHâ)vC(0)N.Rè(OHa)r) “(CHjCHgö}?", -{CHsCH20)rCHri -{CH2}fC{0)NRö(CH*CHgÖV, ^CHg)rC(0)NR&amp;(:CH;>CHs0)r-CHr. ~(CHsCH20)rC{0)NRb{CH2CH20)r, *COH2CH20)iO(0)NRîs(CH2CHâO)rCHr és ^CH2CH20)fC(0)NRb(CHs)r mtU választunk; ahol Rb jelentés© H, Οι·Ό§ aíkií, fenil vagy benzil; r jelentése, függetlenöl, egész szám, értéke 1 ~ 10, 25. A 24, Igénypont szerinti antitest-hatóanyag konjugáturo vegyulet, ahol Ww jelentése vailn-dtruin, 26. A 24, igénypont szerinti antitest-hatóanyag konjugátum vegyület, ahol RK? jelentése (CH2)s vagy (CHg)2. 27. A 24. igénypont szerinti antiteshhaíöanyag konjugátum vegyület, amelynek képlete a következő:
    28. A 27. igénypont szerinti antítest-hatóanyag konjugátum vegyület ahol R17 jelentése (CHájs vagy (CH2}2. 28, A 24, Igénypont szerinti antitest-hatóanyag: konjugátum vegyület, amelynek képlete a követkézé;
    30« Ä 17, Igénypont szerinti antitest-hatóanyag konjugiium vegyülei ahol m L az SMCO vagy a BMFEO kapcsoioreagensbői késiül, 31, A 17, Igénypont szerintl antitest-hatóanyag konjugátum vágyóiéi ahol a D hatóanyagmolekulárészt niikrotybutin-inhiPitöf, mitozls-inhibitor, topöizomefáz-lnhibitor é$ DNSantarkaléió szar közül választjuk, 32, Â 17, igénypont szerinti antitest-hatóanyag konjngáturn vagyaiét, ahol a D hatöanyagmotékuiafészt egy majtanzinoick egy aurlaztatln, egy óoiasziatln és agy kalikearnldn közül választjuk, 33» A 17, igénypont szerinti antitest-hatóanyag konjugátum vágyóiéi ahol D jelentése yfvlAÉ, amelynek szerkezete a következő:
    áhöl a hullámos vonal az L linkerhaz való kapesölóöás helyét jelöli. 34, A 17, igénypont szerinti antitest-hatóanyag konjugátum vegyüiet, ahoi D jelentése MíVt&amp;P, amelynek szerkezete a következő:
    ahol a hullámos vonal az L linkéihez való kapcsolódás helyét jelűé 3§, A 17. Igénypont szerinti antitest-hatóanyag konjugátum vég y Ölet, ahol D jelentése DM1 amelynek szerkezete a kővetkező;
    vagy DM4: amelynek szerkezete a következő;
    ahol a hullámos vonal az L linkerhez való kapcsolódás helyét jelöli. 3β, A 17. igénypont szerinti antitesí~hatóanyag konjugáturn vegyület ahoi a kiindulási antitestet niönoklonälls antitest, hispeoifikus antitest és aníitestfragmens közöl választjuk. 37. .4 17. igénypont, szerinti antitest-hatóanyag konjugáturn vegyület ahol a kiindulási antitestet huMAP4D5~3 .(traszíuzumsb)» anti-€rbB2R antitest« antk EphB2R antitest. anti~CD22 antitest és ant^M'UO-16 antitest közöl választjuk, M. A 17, igénypont szerinti antitest-hatóanyag konjugáíum vegyülök ahol a kiindulási antitest IgA, igD, IgE, IgG és így közűi választott intakt antitest, 3§* A 38. igénypont szerinti antitest-őrig konjugáturn vegyület, ahol az IgG-t a következő .alosztályok közül választjuk: IgGI s igG2* lgG3 és igG4, 40, A 17. igénypont szerinti antitest-hatóanyag konjugáturn vegyület, amelynek szerkezeté a következő:
    ahol n értéke 0S 1 vagy 2; Ah jelentése eisztein-manipulalf antitest, 41, A 17, igénypont szerinti antitest-hatóanyag konjugáturn vegyület, amelynek szerkezetét a következők közül választjuk:
    ahol Val jelentése várni: és Ült Jelentése cltriJin.
  5. 42, Gyógyászati készítmény, amely tartalmazza a 17-41, Igénypontok bármelyik© szerinti antitest-hatóanyag konjugáturn vagy 0 letet vagy annak gyógyászati lag elfogadható sóját, továbbá gyógyászattlsg elfogadható hlgltósiert. hordozót vagy exoipienst, 43. A 42, igénypont szerinti gyógyászati készítmény, amely további kemoterápiás ágens terápiásán hatásos mennyiségét Is tartalmazza. 44. A 17-41, igénypontok bármelyike szerinti antitest-hatóanyag konjugâturn vagy ü leinek vagy gyogyászatilag elfogadható séfénél alkalmazása rák kezelésére alkalmazható gyógyszer előállításában. 4Μ.-Ά 17-41. igénypontok bármelyike szerinti antitest- ha tóanyag konjugâturn vegyidet vagy gyógyászaiilag elfogadható slja rák kezelésire szolgáló éfféráshan történd alkalmazásra.
  6. 45. Eljárás cisztaln-manipulálí antitestek tioreaktivítás szerinti szűrésére, amelynek forán: (a) beviszünk egy vagy több oisztein aminosavat egy kiindulási antitestbe annak érdekében, hogy egy oíszteln-mahiputáit antisstet generálnnkf (b) reagélfafjuk a olszteín-manipuléit antitestet tiolreaktiv atíinliásreageossef, affiniiásjeiOít, aszteín-manipulált antitest előállításához, ahol a iiofreakfv affinitásreagens tartalmaz biotin molekularészletet; és (h) megmérjük az aftlnitásjelőli ciszteln-manipulált antitest kötődését egy kifogó közeghez, ahol a kifogó közeg tartalmaz sztreptavidint; és az eljárás során további a tiolreaktiv reagenssel meghatározzuk a cisztein-manipulált a ntitost tiolreáktlviíléát is. 47. A 46. igénypont szerinti szűrést eljárás, ahol az eljárás (a) lépése tartalmaz: (i) a cl szíe I n~m a n i p u Iá if a nt if estet kódoló gén helyiránystott mutagenezisét egy kéttossziiü fos) piazmidbsn; és 01} a óisztein-manipiáit antitest expresszáliafásál 4i> A 47 igénypont szerinti szűrési eljárás, amelynek során izoláljuk és tisztítjuk Is az expresszáit cisztein-manipulált antitestei 49. A 46. igénypont szerinti szűrásl eljárás, ahol a tiolreaktiv reagens tartalmaz ffiáleiffiid molekularészletét. SS* A 46. igénypont szerinti szűrési eljárás, ahol a kiindulási antitest fúziós fehérje, amely tartalmaz albunankdíő peptide! (ABP) . S1, Az 50. igénypont szerinti szűrési eljárás, ahol m ABP tartalmaz a következük közöl választott, szekvenciát: SEQ ID MO: 1. SEQ ID NO: :2., SEQ ID MO; 3, SEQ ID NO; 4 és SEO ID NO: 5.. 52« A 46, Igénypont szenet! szűrési eljárás, ahol a kiindulási antitest egy Fah antitestfragmens.
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