US2017119903A1PendingUtilityA1
Modified antibody in which motif comprising cysteine residue is bound, modified antibody-drug conjugate comprising the modified antibody, and production method for same
Est. expiryFeb 24, 2032(~5.6 yrs left)· nominal 20-yr term from priority
A61K 47/6871A61P 35/00C07K 2317/73A61K 47/6809A61K 31/704C07K 16/00A61K 47/6851A61K 38/07A61K 47/6817A61K 47/6855C07K 2317/52C07K 16/32A61K 39/395C07K 2317/76C07K 2317/24C07K 16/28A61K 47/6849A61K 47/42C07K 16/30C07K 16/3015A61K 47/65A61P 43/00A61K 47/50A61K 47/48584A61K 47/68031
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Claims
Abstract
The present invention relates to an antibody in which a motif composed of an amino acid or peptide sequence including one or more cysteine residues is bound to the terminus of a parent antibody, particularly the terminus of the heavy chain of the parent antibody. Also, the present invention relates to a modified antibody-drug conjugate (mADC) comprising a drug bound to the antibody, and a method for producing the antibody or the modified antibody-drug conjugate. The modified antibody-drug conjugate according to the invention can accurately deliver the drug to a target cell due its high specificity to antigen, and thus can increase the therapeutic effect of the drug. Also, it can increase the usability of drugs, particularly anticancer drugs, the use of which is restricted due to their toxicity, despite their high efficacy. Moreover, the invention relates to a composition for treatment of diseases, particularly cancers, which comprise the modified antibody-drug conjugate.
Claims
exact text as granted — not AI-modified1 . An antibody-drug conjugate comprising a modified antibody comprising a cysteine-containing motif at the terminus of a parent antibody, with a formula 1:
Xa-[(M Cys ) n -Xb n ] n Formula (1)
wherein (M Cys ) n is a metal ion binding motif that contains a cysteine residue; Xa and Xb n each independently is a peptide comprising 0 to 20 amino acid residues other than cysteine; and n is an integer ranging from 1 to 20,
the metal ion binding motif that contains a cysteine residue is any one or more selected from the group consisting of:
(A) a zinc finger motif that contains C 2 H 2 group (Cys 2 His 2 class: Cys-X 2-4 -Cys-X 12 -His-X 3-5 -His), C 4 group (C4 class: Cys-X 2 -Cys-X m -Cys-X 2 -Cys-X m -Cys-X 2 -Cys-X m -Cys-X 2 -Cys) or C 6 group (C6 class: Cys-X 2 -Cys-X 6 -Cys-X 5-12 -Cys-X 2 -Cys-X 6-8 -Cys) of zinc finger protein, wherein each X is independently an amino acid residue other than Cys; m is an integer ranging from 1 to 10; and X m or X m-q of X 2-4 , X 3-5 , X 5-12 or X 6-8 in the C 2 H 2 group, C 4 group, or C 6 group, is m or m to q amino acid residues other than Cys;
(B) a metal ion binding motif artificially designed based on a zinc finger motif or a metal ion binding protein motif;
(C) a Cys-X m -Cys motif, a Met-X-Cys-X-X-Cys motif, or a C-Q-C-Q-C-A-C motif of transcription regulatory proteins, metal chaperone proteins, metal ion transporters, or superoxide dismutase; wherein each X is independently an amino acid residue other than Cys; m is an integer ranging from 1 to 10;
(D) a Ser-Pro-Cys motif of membrane protein ATPase; and
(E) a metal ion binding peptide motif having a circular structure; and
the drug is conjugated to the thiol group of the cysteine residue in the metal ion binding motif.
2 . The antibody-drug conjugate of claim 1 , wherein the cysteine-containing motif is bound to the terminus of the heavy chain of the parent antibody.
3 . The antibody-drug conjugate of claim 2 , wherein the cysteine-containing motif is bound to the C-terminus of the heavy chain of the parent antibody.
4 . The antibody-drug conjugate of claim 1 , comprising the metal ion binding motif (A), wherein the C 2 H 2 group, the C 4 group or the C 6 group of the zinc finger protein is any one selected from the group consisting of
(SEQ ID NO: 1)
YKCKQCGKAFGCPSNLRRHGRTH,
(SEQ ID NO: 2)
YQCNICGGKCFSCNSNLHRHQRTH,
(SEQ ID NO: 3)
YSCGICGKSFSDSSAKRRHCILH,
(SEQ ID NO: 4)
YTCSDCGKAFRDKSCLNRHRRTH,
(SEQ ID NO: 5)
YRCKYCDRSFSDSSNLQRHVRNIH,
(SEQ ID NO: 6)
YKCKECGKAFNHSSNFNKHHRIH,
(SEQ ID NO: 7)
FKCPVCGKAFRHSSSLVRHQRTH,
(SEQ ID NO: 8)
YRCKYCCDRSFSISSNLQRHVRNIH,
(SEQ ID NO: 9)
YECDHCGKAFSIGSNLNVHRRIH,
(SEQ ID NO: 10)
YGCHLCCKAFSKSSNLRRHEMIH,
(SEQ ID NO: 11)
YKCKECGQAFRQRAHLIRFIHKLH,
(SEQ ID NO: 12)
YKCHQCGKAFIQSFNLRRHERTH,
(SEQ ID NO: 13)
FQCNQCGASFTQKGNLNRHIKLH,
(SEQ ID NO: 14)
YTCSYCGKSFTQSNTLKQHTRIH,
(SEQ ID NO: 15)
YACHLCGKAFTQSSHRRHEKTH,
(SEQ ID NO: 16)
YKCGQCGKFYSQVSHLTRHQKIH,
(SEQ ID NO: 17)
YACHLCGKAFTQCSHLRRHEKTH,
(SEQ ID NO: 18)
YACHLCAKAFIQCSHLRRHEKTH,
(SEQ ID NO: 19)
YVCRECGRGFRQHSHLVRHKRTH,
(SEQ ID NO: 20)
YKCEECEGKAFRQSSHLTTHKIIH,
(SEQ ID NO: 21)
YECDHCGKSFSQSSHLNVHKRTH,
(SEQ ID NO: 22)
YMCSECGRGFSQKSNLTIHQRTH,
(SEQ ID NO: 23)
YKCEECGKAFTQSSNLTKHKKIH,
(SEQ ID NO: 24)
FECKDCGKAFIQKSNLIRHQRTH,
(SEQ ID NO: 25)
YVCRECRRGFSQKSNLIRHQRTH,
(SEQ ID NO: 26)
YECEKCGKAFNQSSNLTRHKKSH,
(SEQ ID NO: 27)
YECVQCGKSYSQSSNLFRHQRRH,
(SEQ ID NO: 28)
YECVQCGKGFTQSSNLITHQRVH,
(SEQ ID NO: 29)
YECNTCRKTFSQKSNLIVHQRTH,
(SEQ ID NO: 30)
YVCSKCGKAFTQSSNLTVHQKIH,
(SEQ ID NO: 31)
YKCDECGKNFTQSSNLIVHKRIH,
(SEQ ID NO: 32)
YECDVCGKTFTQKSNLGVHQRTH,
(SEQ ID NO: 33)
YKCPDCGKSFSQSSSLIRHQRTH,
(SEQ ID NO: 34)
YECQDCGRAFNQNSSLGRHKRTH,
(SEQ ID NO: 35)
YECNECGKFFSQSSSLIRHRRSH,
(SEQ ID NO: 36)
YKCEECGKAFNQSSTLTRHKIVH,
(SEQ ID NO: 37)
YECNECGKAFAQNSTLRVHQRIH,
(SEQ ID NO: 38)
YEVHDCGKSFRQSTHTLTQHRRIH,
(SEQ ID NO: 39)
YECHDCGKSFRQSTHLTRHRRIH,
(SEQ ID NO: 40)
HKCLECGKCFSQNTHLTRHQRTH,
(SEQ ID NO: 41)
YVCDVEGCTWKFARSDELNRHKKRH,
(SEQ ID NO: 42)
YHCDWDGCGWKFARSDELTRHYRKH,
(SEQ ID NO :43)
YRCSWEGCEWRFARSDELTRHFRKH,
(SEQ ID NO: 44)
FSCSWKGCERRFARSDELSRHRRTH,
(SEQ ID NO: 45)
FACSWQDCNKKFARSDELARHYRTH,
(SEQ ID NO: 46)
YHCNWDGCGWKFARSDELTRHYRKH,
(SEQ ID NO: 47)
FLCQYCAQRFGRKDHLTRHMKHSH,
(SEQ ID NO: 48)
CRCNECGKSFSRRDHLVRHQRTH,
(SEQ ID NO: 49)
FQCKTCQRKFSRSDHLKTHTRTH,
(SEQ ID NO: 50)
FACEVCGVRFTRNDKLKIHMRKH,
(SEQ ID NO: 51)
YVCDVEGCTWKFARSDKLNRHKKRH,
(SEQ ID NO: 52)
YKCMECGKAFNRRSHLTRHQRIH,
(SEQ ID NO: 53)
YICRKCGRGFSRKSNLIRHQRTH,
(SEQ ID NO: 54)
YECKECGKAFSSGSNFTRHQRIH,
(SEQ ID NO: 55)
FHCGYCEKSFSVKDYLTKHIRTH,
(SEQ ID NO: 56)
YECDHCGKAFSVSSNLNVHRRIH,
(SEQ ID NO: 57)
YTCKQCGKAFSVSSSLRRHETTH,
(SEQ ID NO: 58)
YECNYCGKTFSVSSTLIRHQRIH,
(SEQ ID NO: 59)
YRCEECGKAFRWPSNLTRHKRIH.
(SEQ ID NO: 60)
FACDICGRKFARSDERKRHTKIH,
(SEQ ID NO: 61)
CPVESCDRRFSRSDELTRHIRIH,
and
(SEQ ID NO: 62)
CDICGRKFARSDERKRHTKIH.
5 . The antibody-drug conjugate of claim 1 , comprising the metal ion binding motif (B), wherein the metal ion binding motif artificially designed based on a zinc finger motif or the metal ion binding protein motif is any one selected from the group consisting of
(SEQ ID NO: 86)
PYKCPECGKSFSQKSALVKHQRTHTC,
(SEQ ID NO: 87)
PYKCPECGKSFSQKSALVKHQRTHTM,
(SEQ ID NO: 88)
PYKCPECGKSFSQKSALVKHQRTHT(Me-C),
(SEQ ID NO: 89)
PYK(Me-C)PECGKSFSQKSALVKHQRTHTH,
(SEQ ID NO: 90)
PYKCPE(Me-C)GKSFSQKSALVKHQRTHTH,
(SEQ ID NO: 91)
PYKCPE(Me-C)GKSFSQKSALVKHQR,
(SEQ ID NO: 92)
GLKALEEKCKALEEKLKALEEKLKALEEKG,
(SEQ ID NO: 93)
GLKALEEKLKALEEKLKACEEKLKALEEKG,
(SEQ ID NO: 94)
GLKALEEKCKALEEKLKACEEKLKALEEKG,
(SEQ ID NO: 95)
GLKALEEKLKALEEKCKALEEKLKALEEKG,
(SEQ ID NO: 96)
GLKALEEKLKALEEKLKALEEKCKALEEKG,
(SEQ ID NO: 97)
GLKALEEKLKALEEKLKALEEKLKAAEEKCKALEEKG,
(SEQ ID NO: 98)
GLKALEEKLKALEEKCKALEEKLKAAEEKCKALEEKG,
(SEQ ID NO: 99)
ELYALEKELGALEKELACLEKELGALEKELYALEK,
(SEQ ID NO: 100)
KLYALKEKLGALKEKLACLKEKLGALKEKLYALKE,
(SEQ ID NO: 101)
ELYALEKELGALEKELACLKEKLGALKEKLYALKE,
and
(SEQ ID NO: 102)
KLYALKEKLGALKEKLACLEKELGALEKELYALEK.
6 . The antibody-drug conjugate of claim 1 , comprising the metal ion binding motif (C), wherein the Cys-X m -Cys motif, the Met-X-Cys-X-X-Cys motif, or the C-Q-C-Q-C-A-C motif of the transcription regulatory proteins, the metal chaperone proteins, the metal ion transporters, or the superoxide dismutase is any one selected from the group consisting of
CEIFCYDEEKVNRIQGDLQTVDISGVSQILKAIADENRAKITYALCQDEELCVC (SEQ ID NO: 63), CDTHLVHLDNVRSSQAQILPTDKAQQMAEIFGVLADTNRIRLLSALASSELCVC (SEQ ID NO: 64)), CDQPLVHLEQVRQVQPEVMSLDQAQQMAEFFSALADPSRLRLMSALARQELCV C (SEQ ID NO: 65), CDRAHLVDCSRVGDIQTQVLNTAKAQRMAEFFSLLGDANRLRVVSVLAKQELC VC (SEQ ID NO: 66), LSDETRLGIVLLLREMGELCVCDLCM (SEQ ID NO: 67), CCTLATGPLSSDESEHYADLFKVLGDPVRLRILSQLAAGGC (SEQ ID NO: 68), YRAAMPVVRALVAYLTENCCHGTRDC (SEQ ID NO: 69), CLRGCGLVVATYEGRQVRYALADSHLARALGELVQVVLAVDTDQPC (SEQ ID NO: 70), CLRDCGLVVTVPDGRRSRYELADERLGHALDDLRAAVVAVDADRTCPDADELE CC (SEQ ID NO: 71), HXXWFYLX 21-28 CXLFMVIGXWFLVIX 18-27 HXXH, wherein X is any amino acid, and X m-q is amino acid residues other than Cys, the number of which is indicated by m to q,
(SEQ ID NO: 72)
VSGMDCAACARKVENAVRQLAGVNQVQVLFA,
(SEQ ID NO: 73)
VPGMTCSACPITVKKAISEVEGVSKVDVTFE,
(SEQ ID NO: 74)
ITGMTCDSCAAHVKEALEKVPGVQSALVSY,
(SEQ ID NO: 75)
VQGFTCANCAGKFEKNVKKIPGVQDAKVNFG,
(SEQ ID NO: 76)
VEGMTCNSCVWTIEQQIGKVNGEHHIKVSLE,
(SEQ ID NO: 77)
VVMTCSGCSGAVNKVLTKLEPDVSKIDIS,
(SEQ ID NO: 78)
GMTCASCVANIERNLRREEGIYSV,
(SEQ ID NO: 79)
YEGMTCQSCVSSIEGKYRKLQGVVRYKVSL,
(SEQ ID NO: 80)
GMSCQGCAGAVRRVLTKMEGVETFDIDME,
(SEQ ID NO: 81)
VPSITCSHCVDKIEKFVGEIEGVSFIDANVE,
(SEQ ID NO: 82)
VTGMTCAACSNSVEAALMNVNGVDVGGMTCGGCSASVKKLLESQPCVASA
SV,
(SEQ ID NO: 83)
VSGMVCAACSTAVENALLSCSGV,
(SEQ ID NO: 84)
DVGGMTCGGCSASVKKILESQP,
and
(SEQ ID NO: 85)
DVGGMKCGGCVEHVKKILEEQFGVTSAS.
7 . The antibody-drug conjugate of claim 1 , comprising the metal ion binding motif (D).
8 . The antibody-drug conjugate of claim 1 , comprising the metal ion binding motif (E), wherein the metal ion binding peptide motif having the circular structure is Cyclo[K 1,12] (QCGVCGKCIACK).
9 . The antibody-drug conjugate of claim 1 , wherein the parent antibody is one or more selected from the group consisting of a monoclonal antibody, a bispecific antibody, a chimeric antibody, a human antibody, and a humanized antibody.
10 . The antibody-drug conjugate of claim 1 , wherein the parent antibody is one or more selected from the group consisting of IgA, IgD, IgE, IgG, and IgM.
11 . The antibody-drug conjugate of claim 1 , wherein the parent antibody has binding affinity and specificity to cancer-specific antigens, cell surface receptor proteins, cell surface proteins, transmembrane proteins, signaling proteins, cell survival regulators, cell proliferation regulators, molecules associated with tissue development or differentiation, lymphokines, cytokines, molecules involved in cell cycle regulation, molecules involved in vasculogenesis, or molecules associated with angiogenesis.
12 . The antibody-drug conjugate of claim 1 , wherein the parent antibody has binding affinity to one or more proteins selected from the group consisting of:
(1) BMPRIB (bone morphogenetic protein receptor-type IB; (2) E16 (LAT1, SLC7A5); (3) STEAP1 (six transmembrane epithelial antigen of prostate); (4) 0772P (CA125, MUC16); (5) MPF (MPF, MSLN, SMR, megakaryocyte potentiating factor, mesothelin); (6) Napi3b (NAPI-3B, NPTIIb, SLC34A2, solute carrier family 34 (sodium phosphate), member 2, type II sodium-dependent phosphate transporter 3b); (7) Sema 5b (FLJ10372, KIAA1445, Mm.42015, SEMA5B, SEMAG, Semaphorin 5b Hlog, sema domain, seven thrombospondin repeats (type 1 and type 1-like), transmembrane domain (TM) and short cytoplasmic domain, (semaphorin) 5B); (8) PSCA hlg (2700050C12Rik, C530008016Rik, RIKEN cDNA 2700050C12, RIKEN cDNA 2700050C12); (9) ETBR (Endothelin type B receptor); (10) MSG783 (RNF124, hypothetical protein FLJ20315); (11) STEAP2 (HGNC_8639, IPCA-1, PCANAP1, STAMP1, STEAP2, STMP, prostate cancer associated gene 1, prostate cancer associated protein 1, six-transmembrane epithelial antigen of prostate 2, six-transmembrane prostate protein); (12) TrpM4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel, subfamily M, member 4); (13) CRIPTO (CR, CR1, CRGF, CRIPTO, TDGF1, teratocarcinoma-derived growth factor); (14) CD21 (CR2 (Complement receptor 2) or C3DR (C3d/Epstein Barr virus receptor) or Hs.73792); (15) CD79b (CD79B, CD790, IGb (immunoglobulin-associated beta), B29); (16) FcRH2 (IFGP4, IRTA4, SPAP1A (SH2 domain containing phosphatase anchor protein 1a), SPAP1B, SPAP1C); (17) HER2; (18) ErbB receptor selected from EGFR, HER3, and HER4; (19) NCA; (20) MDP; (21) IL20Rα; (22) Brevican; (23) EphB2R; (24) ASLG659; (25) PSCA; (26)GEDA; (27) BAFF-R (B cell-activating factor receptor, BLyS receptor, BR3); (28) CD22 (B-cell receptor CD22-B isoform); (29) CD79a (CD79A, CD79.alpha., immunoglobulin-associated alpha, a B cell-specific protein that covalently interacts with Ig beta (CD79B) and forms a complex on the surface with IgM molecules, transduces a signal involved in B-cell differentiation); (30) CXCR5 (Burkitt's lymphoma receptor 1, a G protein-coupled receptor that is activated by the CXCL 13 chemokine, functions in lymphocyte migration and humoral defense, plays a role in HIV-2 infection and regarded for development of AIDS, lymphoma, myeloma, and leukemia); (31) HLA-DOB (Beta subunit of MHC class II molecule (1a antigen) that binds peptides and presents them to CD4+T lymphocytes); (32) P2X5 (Purinergic receptor P2X ligand-gated ion channel 5, an ion channel gated by extracellular ATP, may be involved in synaptic transmission and neurogenesis, deficiency may contribute to the pathophysiology of idiopathic detrusor instability); (33) CD72 (B-cell differentiation antigen CD72, Lyb-2); (34) LY64 (Lymphocyte antigen 64 (RP105), type I membrane protein of the leucine rich repeat (LRR) family, regulates B-cell activation and apoptosis, loss of function is associated with increased disease activity in patients with systemic lupus erythematosis); (35) FcRH1 (Fc receptor-like protein 1, a putative receptor for the immunoglobulin Fc domain that contains C2 type Ig-like and ITAM domains); (36) IRTA2 (Immunoglobulin superfamily receptor translocation associated 2; (37) TENB2 (putative transmembrane proteoglycan); (38) MAGE-C1/CT7; (39) androgen receptor, PTEN, human kallikrein-related peptidase 3; (40) CD20; (41) CD30; (42) CD33; (43) CD52; (44) EpCam; (45) CEA; (46) gpA33; (47) Mucins; (48) TAG-72; (49) Carbonic anhydrase IX; (50) PSMA; (51) folate binding protein; (52) gangliosides (GD2, GD3, GM2); (53) hydrate/saccharide Lewis-Y; (54) VEGF; (55) VEGFR; (56) aVb3; (57) a5b1; (58) ERB3; (59) c-MET; (60) EphA3; (61) TRAIL-R1, TRAIL-R2; (62) RANKL; (63) FAP; and (64) Tenascin.
13 . The antibody-drug conjugate of claim 1 , wherein the parent antibody comprises one or more selected from the group consisting of trastuzumab, rituximab, bevacizumab, cetuximab, panitumumab, ipilimumab, alemtuzumab, ofatumumab, gemtuzumab, brentuximab, 90 Y-ibritumomab, 131 I-tositumomab, cBR96, cAClO, anti-CD20 antibody, anti-EphB2 antibody, anti-IL-8, E-selectin antibody, anti-MUC16 antibody, anti-CD30 antibody, anti-CD33 antibody, and anti-CD52 antibody.
14 . The antibody-drug conjugate of claim 1 , wherein the drug is conjugated to the cysteine residue in the motif by a linker.
15 . The antibody-drug conjugate of claim 14 , wherein the linker is one or more selected from the group consisting of alkyl halide derivatives containing a haloacetyl group, derivatives containing a maleimide group, aryl halide derivatives containing fluorobenzene, aziridine derivatives and acryloyl derivatives, wherein the derivatives comprise an alkylating reactive group, an arylating reactive group, a maleimide group, an aziridine group, an acryloyl group, or a disulfide exchange reactive group comprising pyridyl disulfide or thionitrobenzoic acid, which reacts with the thiol group of the cysteine residue in the motif and is covalently bound thereto.
16 . The antibody-drug conjugate of claim 1 , wherein the drug is one or more selected from the group consisting of microtubulin inhibitors, mitotic inhibitors, topoisomerase inhibitors, or chemotherapeutic agents capable of functioning as DNA intercalators, anticancer agents, protein toxins capable of functioning as enzymes, micro-RNA (miRNA), siRNA, or shRNA, which can inhibit the expression of a specific oncogene, and radioisotopes.
17 . The antibody-drug conjugate of claim 1 , wherein the drug is one or more selected from the group consisting of maytansinoid, auristatin, aminopterin, actinomycin, bleomycin, talisomycin, camptothecin, N 8 -acetyl spermidine, 1-(2 chloroethyl)-1,2-dimethyl sulfonyl hydrazide, taxol, esperamicin, etoposide, 6-mercaptopurine, dolastatin, trichothecene, CC1065, calicheamicin and other enediyne antibiotics, taxane, anthracycline, methotrexate, adriamycin, vindesine, vinca alkaloids, vincristine, vinblastine, etoposide, doxorubicin, melphalan, mitomycin A, mitomycin C, chlorambucil, daunorubicin, daunomycin and stereoisomers, isosteres, or analogs thereof, duocamycin and stereoisomers, isosteres, or analogs thereof, nucleolytic enzymes, antibiotics, toxins of bacterial, plant or animal origin, cisplatin, CPT-11, doxorubicin, paclitaxel, and docetaxel.
18 . The antibody-drug conjugate of claim 1 , wherein the drug is one or more selected from the group consisting of maytansinoid, auristatin, aminopterin, actinomycin, bleomycin, talisomycin, camptothecin, N 8 -acetyl spermidine, 1-(2 chloroethyl)-1,2-dimethyl sulfonyl hydrazide, taxol, esperamicin, etoposide, 6-mercaptopurine, dolastatin, trichothecene, CC1065, calicheamicin and other enediyne antibiotics, taxane, anthracycline, methotrexate, adriamycin, vindesine, vinca alkaloids, vincristine, vinblastine, etoposide, doxorubicin, melphalan, mitomycin A, mitomycin C, chlorambucil, daunorubicin, daunomycin and stereoisomers, isosteres, or analogs thereof, duocamycin and stereoisomers, isosteres, or analogs thereof, nucleolytic enzymes, antibiotics, toxins of bacterial, plant or animal origin, cisplatin, CPT-11, doxorubicin, paclitaxel, and docetaxel.
19 . A therapeutic composition comprising the antibody-drug conjugate according to claim 1 .Cited by (0)
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