CN117467017A - 双特异性四价抗体及其制备和使用方法 - Google Patents
双特异性四价抗体及其制备和使用方法 Download PDFInfo
- Publication number
- CN117467017A CN117467017A CN202311507433.1A CN202311507433A CN117467017A CN 117467017 A CN117467017 A CN 117467017A CN 202311507433 A CN202311507433 A CN 202311507433A CN 117467017 A CN117467017 A CN 117467017A
- Authority
- CN
- China
- Prior art keywords
- domain
- amino acid
- antibody
- acid sequence
- scfv
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000000034 method Methods 0.000 title description 8
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 claims abstract 3
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 claims abstract 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 41
- 101100314454 Caenorhabditis elegans tra-1 gene Proteins 0.000 claims description 6
- HKZAAJSTFUZYTO-LURJTMIESA-N (2s)-2-[[2-[[2-[[2-[(2-aminoacetyl)amino]acetyl]amino]acetyl]amino]acetyl]amino]-3-hydroxypropanoic acid Chemical compound NCC(=O)NCC(=O)NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O HKZAAJSTFUZYTO-LURJTMIESA-N 0.000 claims description 3
- 125000001433 C-terminal amino-acid group Chemical group 0.000 claims description 3
- 210000004027 cell Anatomy 0.000 description 31
- 206010028980 Neoplasm Diseases 0.000 description 23
- 201000011510 cancer Diseases 0.000 description 18
- 239000003814 drug Substances 0.000 description 10
- 150000001413 amino acids Chemical class 0.000 description 9
- 230000000694 effects Effects 0.000 description 9
- 239000012634 fragment Substances 0.000 description 7
- 229940124597 therapeutic agent Drugs 0.000 description 7
- 239000000203 mixture Substances 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 229960000575 trastuzumab Drugs 0.000 description 6
- 108020004414 DNA Proteins 0.000 description 5
- 230000010056 antibody-dependent cellular cytotoxicity Effects 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 239000008194 pharmaceutical composition Substances 0.000 description 5
- 108090000765 processed proteins & peptides Proteins 0.000 description 5
- 230000035755 proliferation Effects 0.000 description 5
- 239000000427 antigen Substances 0.000 description 4
- 102000036639 antigens Human genes 0.000 description 4
- 108091007433 antigens Proteins 0.000 description 4
- 230000009977 dual effect Effects 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 150000007523 nucleic acids Chemical class 0.000 description 4
- 229960002087 pertuzumab Drugs 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108020003175 receptors Proteins 0.000 description 4
- 102000005962 receptors Human genes 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 3
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 3
- 230000001028 anti-proliverative effect Effects 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 210000004899 c-terminal region Anatomy 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 230000004663 cell proliferation Effects 0.000 description 3
- 229940127089 cytotoxic agent Drugs 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000006471 dimerization reaction Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 229940127121 immunoconjugate Drugs 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 230000010474 transient expression Effects 0.000 description 3
- -1 trastuzumab Chemical compound 0.000 description 3
- 239000013598 vector Substances 0.000 description 3
- 206010006187 Breast cancer Diseases 0.000 description 2
- 208000026310 Breast neoplasm Diseases 0.000 description 2
- 208000000461 Esophageal Neoplasms Diseases 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- 206010030155 Oesophageal carcinoma Diseases 0.000 description 2
- 206010033128 Ovarian cancer Diseases 0.000 description 2
- 206010061535 Ovarian neoplasm Diseases 0.000 description 2
- 239000012980 RPMI-1640 medium Substances 0.000 description 2
- 208000005718 Stomach Neoplasms Diseases 0.000 description 2
- NKANXQFJJICGDU-QPLCGJKRSA-N Tamoxifen Chemical compound C=1C=CC=CC=1C(/CC)=C(C=1C=CC(OCCN(C)C)=CC=1)/C1=CC=CC=C1 NKANXQFJJICGDU-QPLCGJKRSA-N 0.000 description 2
- 230000001270 agonistic effect Effects 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 201000004101 esophageal cancer Diseases 0.000 description 2
- 230000001747 exhibiting effect Effects 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 206010017758 gastric cancer Diseases 0.000 description 2
- 230000014509 gene expression Effects 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 229960003881 letrozole Drugs 0.000 description 2
- HPJKCIUCZWXJDR-UHFFFAOYSA-N letrozole Chemical compound C1=CC(C#N)=CC=C1C(N1N=CN=C1)C1=CC=C(C#N)C=C1 HPJKCIUCZWXJDR-UHFFFAOYSA-N 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- 230000019491 signal transduction Effects 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 201000011549 stomach cancer Diseases 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 230000004614 tumor growth Effects 0.000 description 2
- CLPFFLWZZBQMAO-UHFFFAOYSA-N 4-(5,6,7,8-tetrahydroimidazo[1,5-a]pyridin-5-yl)benzonitrile Chemical compound C1=CC(C#N)=CC=C1C1N2C=NC=C2CCC1 CLPFFLWZZBQMAO-UHFFFAOYSA-N 0.000 description 1
- APRZHQXAAWPYHS-UHFFFAOYSA-N 4-[5-[3-(carboxymethoxy)phenyl]-3-(4,5-dimethyl-1,3-thiazol-2-yl)tetrazol-3-ium-2-yl]benzenesulfonate Chemical compound S1C(C)=C(C)N=C1[N+]1=NC(C=2C=C(OCC(O)=O)C=CC=2)=NN1C1=CC=C(S([O-])(=O)=O)C=C1 APRZHQXAAWPYHS-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 108010012934 Albumin-Bound Paclitaxel Proteins 0.000 description 1
- 239000012103 Alexa Fluor 488 Substances 0.000 description 1
- KHOITXIGCFIULA-UHFFFAOYSA-N Alophen Chemical compound C1=CC(OC(=O)C)=CC=C1C(C=1N=CC=CC=1)C1=CC=C(OC(C)=O)C=C1 KHOITXIGCFIULA-UHFFFAOYSA-N 0.000 description 1
- BFYIZQONLCFLEV-DAELLWKTSA-N Aromasine Chemical compound O=C1C=C[C@]2(C)[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CC(=C)C2=C1 BFYIZQONLCFLEV-DAELLWKTSA-N 0.000 description 1
- 102100029822 B- and T-lymphocyte attenuator Human genes 0.000 description 1
- MLDQJTXFUGDVEO-UHFFFAOYSA-N BAY-43-9006 Chemical compound C1=NC(C(=O)NC)=CC(OC=2C=CC(NC(=O)NC=3C=C(C(Cl)=CC=3)C(F)(F)F)=CC=2)=C1 MLDQJTXFUGDVEO-UHFFFAOYSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 102100027207 CD27 antigen Human genes 0.000 description 1
- 101150013553 CD40 gene Proteins 0.000 description 1
- 101100510617 Caenorhabditis elegans sel-8 gene Proteins 0.000 description 1
- GAGWJHPBXLXJQN-UORFTKCHSA-N Capecitabine Chemical compound C1=C(F)C(NC(=O)OCCCCC)=NC(=O)N1[C@H]1[C@H](O)[C@H](O)[C@@H](C)O1 GAGWJHPBXLXJQN-UORFTKCHSA-N 0.000 description 1
- GAGWJHPBXLXJQN-UHFFFAOYSA-N Capecitabine Natural products C1=C(F)C(NC(=O)OCCCCC)=NC(=O)N1C1C(O)C(O)C(C)O1 GAGWJHPBXLXJQN-UHFFFAOYSA-N 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- ZBNZXTGUTAYRHI-UHFFFAOYSA-N Dasatinib Chemical compound C=1C(N2CCN(CCO)CC2)=NC(C)=NC=1NC(S1)=NC=C1C(=O)NC1=C(C)C=CC=C1Cl ZBNZXTGUTAYRHI-UHFFFAOYSA-N 0.000 description 1
- 102000001301 EGF receptor Human genes 0.000 description 1
- 108060006698 EGF receptor Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- VWUXBMIQPBEWFH-WCCTWKNTSA-N Fulvestrant Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3[C@H](CCCCCCCCCS(=O)CCCC(F)(F)C(F)(F)F)CC2=C1 VWUXBMIQPBEWFH-WCCTWKNTSA-N 0.000 description 1
- 208000017891 HER2 positive breast carcinoma Diseases 0.000 description 1
- 102100034458 Hepatitis A virus cellular receptor 2 Human genes 0.000 description 1
- 101000864344 Homo sapiens B- and T-lymphocyte attenuator Proteins 0.000 description 1
- 101000914511 Homo sapiens CD27 antigen Proteins 0.000 description 1
- 101500025419 Homo sapiens Epidermal growth factor Proteins 0.000 description 1
- 101001068133 Homo sapiens Hepatitis A virus cellular receptor 2 Proteins 0.000 description 1
- 101000831007 Homo sapiens T-cell immunoreceptor with Ig and ITIM domains Proteins 0.000 description 1
- 101000801234 Homo sapiens Tumor necrosis factor receptor superfamily member 18 Proteins 0.000 description 1
- 101000955999 Homo sapiens V-set domain-containing T-cell activation inhibitor 1 Proteins 0.000 description 1
- 101000666896 Homo sapiens V-type immunoglobulin domain-containing suppressor of T-cell activation Proteins 0.000 description 1
- 108091006905 Human Serum Albumin Proteins 0.000 description 1
- 102000008100 Human Serum Albumin Human genes 0.000 description 1
- 229940076838 Immune checkpoint inhibitor Drugs 0.000 description 1
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 1
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 239000005517 L01XE01 - Imatinib Substances 0.000 description 1
- 239000005411 L01XE02 - Gefitinib Substances 0.000 description 1
- 239000005551 L01XE03 - Erlotinib Substances 0.000 description 1
- 239000002147 L01XE04 - Sunitinib Substances 0.000 description 1
- 239000005511 L01XE05 - Sorafenib Substances 0.000 description 1
- 239000002067 L01XE06 - Dasatinib Substances 0.000 description 1
- 239000002136 L01XE07 - Lapatinib Substances 0.000 description 1
- 239000005536 L01XE08 - Nilotinib Substances 0.000 description 1
- 239000003798 L01XE11 - Pazopanib Substances 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 108010061593 Member 14 Tumor Necrosis Factor Receptors Proteins 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 101710100969 Receptor tyrosine-protein kinase erbB-3 Proteins 0.000 description 1
- 102100029986 Receptor tyrosine-protein kinase erbB-3 Human genes 0.000 description 1
- 239000012505 Superdex™ Substances 0.000 description 1
- 102100024834 T-cell immunoreceptor with Ig and ITIM domains Human genes 0.000 description 1
- 102100028785 Tumor necrosis factor receptor superfamily member 14 Human genes 0.000 description 1
- 102100033728 Tumor necrosis factor receptor superfamily member 18 Human genes 0.000 description 1
- 101710165473 Tumor necrosis factor receptor superfamily member 4 Proteins 0.000 description 1
- 102100022153 Tumor necrosis factor receptor superfamily member 4 Human genes 0.000 description 1
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 description 1
- 102100038929 V-set domain-containing T-cell activation inhibitor 1 Human genes 0.000 description 1
- 102100038282 V-type immunoglobulin domain-containing suppressor of T-cell activation Human genes 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000008351 acetate buffer Substances 0.000 description 1
- 229960001686 afatinib Drugs 0.000 description 1
- ULXXDDBFHOBEHA-CWDCEQMOSA-N afatinib Chemical compound N1=CN=C2C=C(O[C@@H]3COCC3)C(NC(=O)/C=C/CN(C)C)=CC2=C1NC1=CC=C(F)C(Cl)=C1 ULXXDDBFHOBEHA-CWDCEQMOSA-N 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 229960003437 aminoglutethimide Drugs 0.000 description 1
- ROBVIMPUHSLWNV-UHFFFAOYSA-N aminoglutethimide Chemical compound C=1C=C(N)C=CC=1C1(CC)CCC(=O)NC1=O ROBVIMPUHSLWNV-UHFFFAOYSA-N 0.000 description 1
- 229960002932 anastrozole Drugs 0.000 description 1
- YBBLVLTVTVSKRW-UHFFFAOYSA-N anastrozole Chemical compound N#CC(C)(C)C1=CC(C(C)(C#N)C)=CC(CN2N=CN=C2)=C1 YBBLVLTVTVSKRW-UHFFFAOYSA-N 0.000 description 1
- 229940046836 anti-estrogen Drugs 0.000 description 1
- 230000001833 anti-estrogenic effect Effects 0.000 description 1
- 229940124691 antibody therapeutics Drugs 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 208000014581 breast ductal adenocarcinoma Diseases 0.000 description 1
- 201000010983 breast ductal carcinoma Diseases 0.000 description 1
- 230000009702 cancer cell proliferation Effects 0.000 description 1
- 230000005907 cancer growth Effects 0.000 description 1
- 229960004117 capecitabine Drugs 0.000 description 1
- 150000001735 carboxylic acids Chemical class 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000009087 cell motility Effects 0.000 description 1
- 238000001516 cell proliferation assay Methods 0.000 description 1
- 229960004316 cisplatin Drugs 0.000 description 1
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 1
- 239000007979 citrate buffer Substances 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 239000000824 cytostatic agent Substances 0.000 description 1
- 230000001085 cytostatic effect Effects 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 239000002254 cytotoxic agent Substances 0.000 description 1
- 231100000599 cytotoxic agent Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 229960002448 dasatinib Drugs 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 230000000431 effect on proliferation Effects 0.000 description 1
- 229960001433 erlotinib Drugs 0.000 description 1
- AAKJLRGGTJKAMG-UHFFFAOYSA-N erlotinib Chemical compound C=12C=C(OCCOC)C(OCCOC)=CC2=NC=NC=1NC1=CC=CC(C#C)=C1 AAKJLRGGTJKAMG-UHFFFAOYSA-N 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 239000000328 estrogen antagonist Substances 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 229960000255 exemestane Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 229950011548 fadrozole Drugs 0.000 description 1
- IJJVMEJXYNJXOJ-UHFFFAOYSA-N fluquinconazole Chemical compound C=1C=C(Cl)C=C(Cl)C=1N1C(=O)C2=CC(F)=CC=C2N=C1N1C=NC=N1 IJJVMEJXYNJXOJ-UHFFFAOYSA-N 0.000 description 1
- 229960004421 formestane Drugs 0.000 description 1
- OSVMTWJCGUFAOD-KZQROQTASA-N formestane Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CCC2=C1O OSVMTWJCGUFAOD-KZQROQTASA-N 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 229960002258 fulvestrant Drugs 0.000 description 1
- 229960002584 gefitinib Drugs 0.000 description 1
- XGALLCVXEZPNRQ-UHFFFAOYSA-N gefitinib Chemical compound C=12C=C(OCCCN3CCOCC3)C(OC)=CC2=NC=NC=1NC1=CC=C(F)C(Cl)=C1 XGALLCVXEZPNRQ-UHFFFAOYSA-N 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 201000010536 head and neck cancer Diseases 0.000 description 1
- 208000014829 head and neck neoplasm Diseases 0.000 description 1
- 229940116978 human epidermal growth factor Drugs 0.000 description 1
- 229960002411 imatinib Drugs 0.000 description 1
- KTUFNOKKBVMGRW-UHFFFAOYSA-N imatinib Chemical compound C1CN(C)CCN1CC1=CC=C(C(=O)NC=2C=C(NC=3N=C(C=CN=3)C=3C=NC=CC=3)C(C)=CC=2)C=C1 KTUFNOKKBVMGRW-UHFFFAOYSA-N 0.000 description 1
- 239000012274 immune-checkpoint protein inhibitor Substances 0.000 description 1
- 230000016784 immunoglobulin production Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000001361 intraarterial administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 206010073095 invasive ductal breast carcinoma Diseases 0.000 description 1
- 229960004891 lapatinib Drugs 0.000 description 1
- BCFGMOOMADDAQU-UHFFFAOYSA-N lapatinib Chemical compound O1C(CNCCS(=O)(=O)C)=CC=C1C1=CC=C(N=CN=C2NC=3C=C(Cl)C(OCC=4C=C(F)C=CC=4)=CC=3)C2=C1 BCFGMOOMADDAQU-UHFFFAOYSA-N 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 208000037819 metastatic cancer Diseases 0.000 description 1
- 208000011575 metastatic malignant neoplasm Diseases 0.000 description 1
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 1
- GVUGOAYIVIDWIO-UFWWTJHBSA-N nepidermin Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H](CS)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CS)NC(=O)[C@H](C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C(C)C)C(C)C)C1=CC=C(O)C=C1 GVUGOAYIVIDWIO-UFWWTJHBSA-N 0.000 description 1
- 201000002120 neuroendocrine carcinoma Diseases 0.000 description 1
- 229960001346 nilotinib Drugs 0.000 description 1
- HHZIURLSWUIHRB-UHFFFAOYSA-N nilotinib Chemical compound C1=NC(C)=CN1C1=CC(NC(=O)C=2C=C(NC=3N=C(C=CN=3)C=3C=NC=CC=3)C(C)=CC=2)=CC(C(F)(F)F)=C1 HHZIURLSWUIHRB-UHFFFAOYSA-N 0.000 description 1
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 238000011275 oncology therapy Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000002611 ovarian Effects 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 229960000639 pazopanib Drugs 0.000 description 1
- CUIHSIWYWATEQL-UHFFFAOYSA-N pazopanib Chemical compound C1=CC2=C(C)N(C)N=C2C=C1N(C)C(N=1)=CC=NC=1NC1=CC=C(C)C(S(N)(=O)=O)=C1 CUIHSIWYWATEQL-UHFFFAOYSA-N 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 229940068965 polysorbates Drugs 0.000 description 1
- 230000003651 pro-proliferative effect Effects 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 238000009097 single-agent therapy Methods 0.000 description 1
- 238000001542 size-exclusion chromatography Methods 0.000 description 1
- 229960003787 sorafenib Drugs 0.000 description 1
- 229960001796 sunitinib Drugs 0.000 description 1
- WINHZLLDWRZWRT-ATVHPVEESA-N sunitinib Chemical compound CCN(CC)CCNC(=O)C1=C(C)NC(\C=C/2C3=CC(F)=CC=C3NC\2=O)=C1C WINHZLLDWRZWRT-ATVHPVEESA-N 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 229960001603 tamoxifen Drugs 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 229960001771 vorozole Drugs 0.000 description 1
- XLMPPFTZALNBFS-INIZCTEOSA-N vorozole Chemical compound C1([C@@H](C2=CC=C3N=NN(C3=C2)C)N2N=CN=C2)=CC=C(Cl)C=C1 XLMPPFTZALNBFS-INIZCTEOSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
- C07K16/3015—Breast
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
- A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
- A61K39/39558—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against tumor tissues, cells, antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
- A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
- A61K47/6849—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
- A61K47/6851—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
- A61K47/6875—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody being a hybrid immunoglobulin
- A61K47/6879—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody being a hybrid immunoglobulin the immunoglobulin having two or more different antigen-binding sites, e.g. bispecific or multispecific immunoglobulin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/32—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products of oncogenes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/46—Hybrid immunoglobulins
- C07K16/468—Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/515—Animal cells
- A61K2039/5152—Tumor cells
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
- C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
- C07K2317/35—Valency
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
- C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
- C07K2317/622—Single chain antibody (scFv)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
- C07K2317/64—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising a combination of variable region and constant region components
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
- C07K2317/732—Antibody-dependent cellular cytotoxicity [ADCC]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Cell Biology (AREA)
- Oncology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Endocrinology (AREA)
- Biomedical Technology (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
一种双特异性四价抗体,其包含具有一对重链和一对轻链的IgG,并且两个scFv组分连接至重链或轻链的C端或N端。双特异性四价抗体可以对HER2受体上的两种不同表位具有结合特异性。
Description
本申请是2015年12月19日提交的发明名称为“双特异性四价抗体及其制备和使用方法”(申请号:2015800734309)的中国发明专利申请的分案申请。
相关专利申请的交叉引用
本申请要求于2014年12月22日提交的题为“BISPECIFIC ANTIBODIES(双特异性抗体)”的美国第62095348号临时申请的优先权,其通过引用整体并入本文。
技术领域
本公开总体上涉及抗体的技术领域,更具体地涉及双特异性抗体。
背景技术
HER2是ErbB/HER受体家族的成员,在乳腺和卵巢的几种癌症中过表达和/或失调(King,Kraus and Aaronson,Science 1985;229:974-976;Slamon et al.,Science 1989;244:707-712)。HER2靶向治疗药物已经在临床上成功使用,并已被美国FDA批准。这样的抗体治疗药物包括曲妥珠单抗(Horton,Cancer Control 2001:8(1),103-110)和帕妥珠单抗(Badache and Hynes,Cancer Cell;5(4):299-301)。一些研究表明,与单一抗体单药治疗相比,可以通过组合两种或多种表位不同的抗HER2抗体(诸如曲妥珠单抗和帕妥珠单抗)来实现增强治疗(Kasprzyk et al.,Cancer Res 1992;52:2771-2776,Ben-Kasus et al.,Proc Natl Acad Sci USA,106(9)3294-3329)。结合HER2细胞外结构域4的曲妥珠单抗抑制非配体依赖性信号传导、刺激ADCC、阻断HER2脱落但不抑制HER2二聚化。与细胞外结构域2结合的帕妥珠单抗抑制HER2二聚化和与其他HER家族受体的二聚化,抑制多种配体依赖性HER介导的信号通路并刺激ADCC(O'Sullivan and Connolly,Oncology 2014;28(3):186-194)。
基于单独使用曲妥珠单抗的显著临床益处,FDA已经于2013年批准了曲妥珠单抗和帕妥珠单抗组合治疗HER2阳性转移性癌症作为HER2阳性乳腺癌的新治疗方法,(Baselgaet al.N Engl J Med.2012 Jan 12;366(2):109-19)。然而,使用两种或多种单克隆抗体的简单组合的功效是次优的。此外,单独生产两种或多种单克隆抗体的成本高。
因此,需要通过组合单克隆抗体来改善癌症治疗的功效并降低与单克隆抗体生产相关的成本。
发明内容
本公开提供双特异性四价抗体。双特异性四价抗体可以包括两条IgGl重链;两条kappa轻链;以及两个单链Fv(scFv)结构域。两条IgGl重链和kappa轻链可以形成对HER2的第一结构域具有结合特异性的IgG部分。两个scFv结构域可以对HER2的第二结构域具有结合特异性。IgG部分和两个scFv结构域共价连接,作为双特异性四价抗体起作用。从以下结合附图对其优选实施例的详细描述中,本公开的目的和优点可变得显而易见。
附图说明
现在可以参考附图来描述根据本公开的优选实例,其中相同的附图标记表示相同的元件。
图1示出了根据本发明一实例的四价双特异性抗体结构。
图2示出了根据本发明的实例的示例性四价双特异性抗体4X1、4X2、4X3和4X4的功能框图。
图3示出了示例性单特异性抗体4C1和4C2的功能框图。
图4示出了示例性Fc-scFv抗体4C3、4C4、4C5、4C6、4C7、4C8、4C10和4C11的功能框图。
图5示出了SI-4X和SI-4C抗体对BT-474细胞增殖的作用。
图6示出了将连接体长度从10个氨基酸扩展到30个氨基酸对BT-474细胞增殖的作用。
图7示出了将连接体长度扩展到40个氨基酸对BT-474细胞增殖的作用。
图8示出了SI-4X抗体对BT-474细胞上HER2内化的作用。
具体实施方式
本公开提供了双特异性四价抗体。该抗体可以具有同时靶向HER2的细胞外结构域2和4的优点。
必须注意的是,如本文和所附权利要求中所使用的,除非上下文另有明确规定,否则在整个本说明书和权利要求书中,单数形式“一”、“和”以及“该”,词语“包含”或诸如“包含”或“包含……的”将被理解为暗示包含所述整数或整数组,但不排除任何其他整数或整数组,包括复数对象。
“抗体片段”包含完整抗体的一部分,优选是完整抗体的抗原结合区或可变区。抗体片段的实例包括Fv、Fab、Fab'、F(ab')2、Fab'-SH;双抗体;线性抗体(参见美国专利第5,641,870号,实例2;Zapata et al.,Protein Eng.8(10):1057-1062(1995));单链抗体分子(例如,scFv)。尽管在本说明书中并且在整个说明书中,提及抗体和抗体的各种性质,但是相同的公开内容也适用于功能性抗体片段,例如,双重作用Fab片段。
一方面,双特异性四价抗体可以包括两条IgGl重链;两条kappa轻链;以及两个单链Fv(scFv)结构域。两条IgGl重链和kappa轻链可以形成对HER2的第一结构域具有结合特异性的IgG部分。两个scFv结构域可以对HER2的第二结构域具有结合特异性。每个scFv结构域可以通过具有氨基酸序列(gly-gly-gly-gly-ser)n((G4S)n)的连接体与任一个IgGl重链的C端残基连接。每个scFv结构域可以具有N端-可变重链-接头-可变轻链-C端或N端-可变轻链-接头-可变重链-C端的结构顺序,并且接头可以包括(gly-gly-gly-gly-ser)m((G4S)m)氨基酸序列。n和m均为整数,n可以是至少为2的整数。在一实施例中,n为1至10或2至9。在一些实施例中,n至少为9。在一些实施例中,n为2至20。m可以是至少为2的整数。在一些实施例中,m可以是2、3、4或5。在一些实施例中,m可以是选自2、3、4、5、6、7、8、9和10的整数。
在一些实施例中,IgGl重链或kappa轻链中的至少一个是人源化的或人的。在一些实施例中,两条IgGl重链均是人源化的或人的。在一些实施例中,两条kappa轻链均是人源化的或人的。
在一些实施例中,HER2的结构域独立地选自HER2的结构域2和结构域4。在一些实施例中,双特异性四价抗体可以包括具有经由肽接头连接到重链或轻链的C端或N端的scFv的IgG部分。IgG部分可以对HER2(人表皮生长因子2)表达细胞的细胞外结构域2或4具有结合特异性,而scFv结构域可以分别对HER2表达细胞的胞外域4或2具有结合特异性。结合可以是二价的。
肽接头的长度可能不同。在一些实施例中,肽接头可包括约15至约45个氨基酸。在一些实施例中,肽接头可包括约20至约50个氨基酸。在一些实施例中,肽接头可包括约10至约30个氨基酸。
在一些实施例中,IgG部分可以对HER2的结构域2具有结合特异性。在一些实施例中,scFv结构域可以对HER2的结构域4具有结合特异性。在一些实施例中,IgG部分可以对HER2的结构域4具有结合特异性。在一些实施例中,scFv结构域可以对HER2的结构域2具有结合特异性。
在一些实施例中,IgGl重链中的至少一个或两个包含SEQ ID NO 7、15、30、40、50和58的氨基酸序列或与SEQ ID NO 7、15、30、40、50和58具有至少95%、98%或99%相似性的氨基酸序列。在一些实施例中,IgGl重链、连接体以及scFv结构域具有SEQ ID NO 30、50、40和58的氨基酸序列或与SEQ ID NO 30、50、40和58具有至少95%、98%或99%相似性的氨基酸序列。在一些实施例中,kappa轻链中的至少一个或两个包含SEQ ID NO 3、11、25、35、45和53的氨基酸序列或与SEQ ID NO 3、11、25、35、45和53具有至少95%、98%或99%相似性的氨基酸序列。在一些实施例中,可变轻链中的至少一个或两个包含SEQ ID NO 4、12、26、36、46和54的氨基酸序列或与SEQ ID NO 4、12、26、36、46和54具有至少95%、98%或99%相似性的氨基酸序列。在一些实施例中,可变重链中的至少一个或两个包含SEQ ID NO8、16、31、41、79和59的氨基酸序列或与SEQ ID NO 8、16、31、41、79和59具有至少95%、98%或99%相似性的氨基酸序列。在一些实施例中,至少一个或两个scFv结构域包含SEQ ID NO19、22、32、42、80、60、63、66、69、72、75、78的氨基酸序列或与SEQ ID NO 19、22、32、42、80、60、63、66、69、72、75、78具有至少95%、98%或99%相似性的氨基酸序列。
在一些实施例中,IgG部分可以对HER2的结构域2具有结合特异性,并且scFv结构域可以对HER2的结构域4具有结合特异性。在一实施例中,IgGl重链、连接体以及scFv结构域可以具有SEQ ID NO 30的氨基酸序列或与SEQ ID NO 30具有至少95%相似性的氨基酸序列,并且kappa轻链可以具有SEQ ID NO 25的氨基酸序列或与SEQ ID NO 25具有至少95%相似性的氨基酸序列。在一实施例中,IgGl重链、连接体以及scFv结构域可以具有SEQID NO 50的氨基酸序列或与SEQ ID NO 50具有至少95%相似性的氨基酸序列,并且kappa轻链可以具有SEQ ID NO 45的氨基酸序列或与SEQ ID NO 45具有至少95%相似性的氨基酸序列。
在一些实施例中,IgG部分可以对HER2的结构域4具有结合特异性,并且scFv结构域可以对HER2的结构域2具有结合特异性。在一实施例中,IgGl重链、连接体以及scFv结构域可以具有SEQ ID NO 40的氨基酸序列或与SEQ ID NO 40具有至少95%相似性的氨基酸序列,并且kappa轻链可以具有SEQ ID NO 35的氨基酸序列或与SEQ ID NO 35具有至少95%相似性的氨基酸序列。在一实施例中,IgGl重链、连接体以及scFv结构域可以具有SEQID NO 58的氨基酸序列或与SEQ ID NO 58具有至少95%相似性的氨基酸序列,并且kappa轻链可以具有SEQ ID NO 53的氨基酸序列或与SEQ ID NO 53具有至少95%相似性的氨基酸序列。
双特异性四价抗体具有抑制癌细胞生长的活性。在某些实施例中,本发明的抗体对于其靶标EGRF或HER3的解离常数(Kd)≤80nM、≤50nM、≤30nM、≤20nM、≤10nM或≤0.1nM。抗体可以同时与两个靶标结合。在一些实施例中,抗体可以以小于1nM、10nM、20nM、50nM或100nM的Kd与HER2的结构域2结合。在一些实施例中,抗体可以以小于5nM、10nM、20nM、50nM或100nM的Kd与HER2的结构域4结合。在一些实施例中,抗体可以以小于30nM的Kd与HER2的结构域4结合,并以小于30nM的Kd与HER2的结构域2结合。在一些实施例中,抗体可以以小于50nM的Kd与HER2的结构域4结合,并同时以小于20nM的Kd与HER2的结构域2结合。
在一些实施例中,IgG部分可以为scFv结构域提供稳定性。另外可选地,IgG部分可以提供对表位的特异性。在一些实施例中,双特异性抗体可介导对表达HER2的细胞的ADCC(抗体依赖性细胞介导的细胞毒性)。在一些实施例中,抗体可能能够结合HER2抗原上的至少两个结构域(即表位)。在一些实施例中,抗体可以同时结合HER2抗原上的多个结构域。
在一些实施例中,抗体可以在体外和体内增殖试验中提供比单特异性抗体亲本对照或单特异性抗体亲本对照的组合更强的肿瘤抑制。不希望被理论限制,据信通过对两种不同表位的相同抗原作用,本文公开的双特异性四价抗体可以增强受体(HER2)的内化,并且比单独单特异性抗体或两种单特异性抗体的组合之一更有效地下调信号通路。
在一些实施例中,双特异性四价抗体可抑制癌细胞生长。在一些实施例中,癌细胞可以表达HER2。在一些实施例中,双特异性四价抗体可抑制癌细胞生长。在一些实施例中,癌细胞可以表达HER2+。
在另一方面,本公开提供了编码本文公开的双特异性四价抗体、片段或亚组分的分离的核酸。
在另一方面,本公开提供了具有编码本文公开的双特异性四价抗体、片段或亚组分的分离的核酸的表达载体。载体可在宿主细胞中表达。宿主细胞可以是原核的或真核的。
在另一方面,本公开提供具有本文公开的双特异性四价抗体、片段或亚组分的分离的核酸的宿主细胞,或包括该核酸序列的表达载体。
在另一方面,本公开提供了制备双特异性四价抗体的方法。在一实施例中,该方法可以包括培养上述宿主细胞,使得产生抗体。
在另一方面,本公开提供了免疫交联物,包括本文所述的双特异性四价抗体和细胞毒性剂。
在另一方面,本公开提供了药物组合物。药物组合物可以包括双特异性四价抗体或本文所述的免疫交联物和药学上可接受的载体。在一些实施例中,组合物还可包括放射性同位素、放射性核素、毒素、治疗剂、化学治疗剂或其组合。
在另一方面,本公开提供了治疗癌症患者的方法。在一实施例中,该方法包括对患者施用有效量的本文所述的双特异性四价抗体的步骤。癌症可以包括表达HER2、结构域、表位、片段或其衍生物的细胞。癌症可能是HER2+乳腺癌、结肠直肠癌、卵巢癌、胃癌、食管癌、头颈癌以及非小细胞肺癌。
在一实施例中,该方法还可包括共同施用有效量的治疗剂。治疗剂可以是例如抗体、化疗剂、酶或其组合。在一些实施例中,治疗剂可以是抗雌激素剂,受体酪氨酸抑制剂或其组合。在一些实施例中,治疗剂可以是卡培他滨、顺铂、曲妥珠单抗、氟维司群、他莫昔芬、来曲唑、依西美坦、阿那曲唑、氨鲁米特、睾丸酯、伏氯唑、福美斯坦、法倔唑、来曲唑、厄洛替尼、阿法替尼、达沙替尼、吉非替尼、伊马替尼、帕唑帕尼、拉帕替尼、舒尼替尼、尼罗替尼、索拉非尼、白蛋白结合型紫杉醇。在一些实施例中,需要这种治疗的患者是人。在一些实施例中,治疗剂可以是生物制剂。在一些实施例中,治疗剂可以是检查点抑制剂,包括但不限于PDl、PDLl、CTLA4、4-lBB、OX40、GITR、TIM3、LAG 3、TIGIT、CD40、CD27、HVEM、BTLA、VISTA和B7H4。
在一实施例中,本公开提供了通过向患者施用有效量的双特异性四价抗体以抑制HER2受体的生物学活性来治疗患者的方法。
在一实施例中,本公开提供了具有有效浓度的双特异性四价抗体的溶液。在一实施例中,该溶液是患者中的血浆。
双特异性四价抗体的一般结构示意图如图1所示。在一实施例中,双特异性四价抗体包括两条人IgGl重链、两条人kappa轻链以及两个单链Fv(scFv)结构域。两条人IgGl重链和人kappa轻链形成IgG部分。两个scFv结构域分别连接到具有连接体的人IgGl重链的C端残基,该连接体具有gly-gly-gly-gly-ser-重复(也被称为(G4S)n)的氨基酸序列。n可以是整数。在一实施例中,n为2至10。在一些实施例中,n可以是1至15。scFv可以为顺序:N端-可变重链-接头-可变轻链-C端。scFv接头可以包括Gly-gly-gly-gly-ser重复(也被称为(G4S)m)的氨基酸序列。m为整数。例如,m可以是3或4。对于所有构建体,CHI、CH2、CH3和CL氨基酸序列可以相同。有4个双特异性抗体,称为4X1、4X2、4X3和4X4。这些在图2中示出。
每个双特异性四价抗体可以在一端与HER2的细胞外结构域2特异性结合,在另一端与HER2的细胞外结构域4特异性结合。这2种抗-HER2结合结构域分别称为4C1和4C2。结构4X1在双特异性抗体的氨基末端具有常规IgGl/kappa重链和轻链形式的4C1结合结构域,在羧基末端加入4C2作为单链Fv。4X2方向相反,4C2位于氨基末端,4C1为羧基末端单链Fv。存在可以使用这些结合对创建的多种其他类型的双特异性抗体结构,包括接头和连接体序列的改变以及这些结合结构域的可选位置和/或形式。例如,通过将4X1的连接体从(G4S)x2扩展到(G4S)x6可以创建4X3,并且可以通过将4X2的连接体从(G4S)x2扩展到(G4S)x6来创建4X4。
为了研究连接体长度的作用,已经生成了称为4C3、4C4、4C5、4C6、4C7、4C8、4C10和4C11的多个Fc-scFv构建体。4C3包含来自4C1的scFv,而4C4包含来自4C2的scFv。为4C4生成(G4S)x3至(G4S)x8的连接体变体,如图4所示,例如,4C5具有15个氨基酸(G4S)x3的连接体长度,而4C11具有40个氨基酸(G4S)x8的连接体长度。表1示出了不同变体的连接体长度。
表1.变体名称和连接体长度
可变轻链,可变重链和单链Fv(scFv)DNA片段通过基因合成生成。通过基因合成生成人Gamma-1重链和人kappa轻链DNA片段。通过使用限制性位点的DNA连接将片段组装在一起,并克隆到设计用于哺乳动物细胞中瞬时表达的载体中。载体含有强CMV衍生的启动子,以及瞬时表达所需的其他上游和下游元件。通过DNA测序验证所得IgG表达质粒含有预期的DNA序列。按CSH手册所述(CSH Protocols;2008;doi:10.1101/pdb.prot4977),用线性PEI转染悬浮适应的HEK293F细胞来实现抗体构建体的瞬时表达。简言之,将DNA加入每个含有F17表达培养基的管中,该培养基已在37℃预热,然后加入PEI。在室温下孵育15分钟,并将DNA/PEI混合物加入含有HEK293细胞(在F17完全培养基中HEK293细胞密度为约1x106细胞/ml)的烧瓶中。在37℃下振荡孵育5天,然后将样品离心,收集上清液并在4℃下储存以进行纯化。
需要时使用蛋白质亲和层析和尺寸排阻色谱法从所得转染上清液中纯化抗体。蛋白质质量用Superdex 200柱分析。用于所有试验的蛋白质纯度大于90%。
对HER2两个不同表位特异的双特异性抗体可用于治疗许多HER2表达的癌症,诸如乳腺癌、卵巢癌、胃癌、食道癌、前列腺癌、肺癌以及神经内分泌癌。
在一实施例中,双重特异性抗体具有四价双特异性。其包括IgG和两个scFv,其与单特异性抗体IgG相比提供两种不同的结合特异性。IgG组分提供了优于仅使用scFv(诸如BiTE技术(Lutterbuese et al,Proceedings of the National Academy of Sciences ofthe United States of America 107.28(2010):12605-12610.PMC.Web.2Dec.2014)和其他(美国专利第7332585号))的其他双特异性抗体的稳定性。其也能够介导ADCC,而无Fc组分的那些则不能(美国专利第7332585号)。四价双重特异性性质为双特异性抗体提供了优于一些其他双特异性抗体的同时结合能力,其可以一次仅结合一种抗原(Kontermann,MAbs.2012 Mar-Apr;4(2):182-97;Schanzer et al,Antimicrob.AgentsChemother.2011,55(5):2369;EP272942)。
为了便于叙述,双特异性抗体的序列或与双特异性抗体相关的序列总结在下面的表2中。
表2.双特异性抗体或与之相关的核苷酸和氨基酸序列的总结
/>
实例
实例1
为了评估抗HER2抗体的生长抑制潜力,测试了对作为乳腺导管癌肿瘤细胞系的BT-474细胞(ATCC HTB-20,Manassas,Va.)增殖的作用。将细胞用含有1%胎牛血清的100μlRPMI-1640培养基以6000细胞/孔的密度接种到96孔组织培养板中。4小时后,以0.0061nM至400nM的不同浓度加入试验抗体。在试验抗体存在下培养细胞7天。向每个孔中加入20μlMTS试剂(Promega,Madison,Wl),将细胞在37℃下孵育2小时。MTS容易被活跃增殖的细胞吸收,还原成甲腊(其易吸收490nm的光),然后分泌到培养基中。孵育后,使用BioTek(Winooski,VT)ELx800吸光度读数器测量OD490值。在抗体添加时也以这种方式获得对照细胞(仅用培养基处理)的OD490值,以确立基线代谢活性。可以通过从72小时OD490中减去对照基线OD490来计算增殖。来自抗体滴定的数据根据以下公式以对照群体的百分比表示:对照增殖的百分比=(试验增殖/对照增殖)*100。
SI-4X1和SI-4X2对BT-474增殖的作用如图5所示。这两种分子均具有抗增殖作用,但两者均不如对照抗体SI-4C2或对照抗体SI-4C1和SI-4C2的组合有效。从SI-4X3可以看出,将C端scFv与hulgG分开的连接体G4S接头的长度从2个重复增加到6个重复增加了功效。怀疑双特异性抗体的较低功效可能是C端scFv提供的促增殖活性的结果。之前有抗-Her2抗体显示出依赖其结构的激动活性的文献。为了研究这一点,我们创建了一系列含有相同抗-Her2 scFv的对照分子,但是具有逐渐变长的G4S接头。如图6中所示,抗增殖作用与接头中G4S元件的数量直接成正比,其中SI-4C8(6个重复)显示出最高的抗增殖活性,而SI-4C4(2个重复)则表现出激动活性。当接头增加到7(SI-4C10)和8(SI-4C11)个重复时,该作用甚至更明显,在图7中示出。
实例2
测试抗-Her2抗体被BT-474细胞内化的能力。使1mg等分试样的抗体标准PBS溶液与Alexa Fluor 488羧酸、TFP酯(Thermo Fisher#A-10235,Waltham,MA)在室温下反应1小时。使用Bio-Gel P-30柱通过凝胶过滤除去未结合的染料。偶联后,将3x10 5 BT-474细胞的等分试样在37℃或4℃(冰)下,在完全培养基(RPMI-1640+10%FBS)中与50nM每种Alexa488标记的抗体一起孵育1小时。孵育后,将细胞在冷的离心机中用冰冷的PBS洗涤两次。然后将细胞重悬于500nM淬灭兔-抗-Alexa488抗体(Thermo Fisher#A-11094,Waltham,MA)或500nM兔IgG同种型对照抗体(Jackson ImmunoResearch Laboratories#011-000-003,WestGrove,PA)任一者中,并在冰上孵育30分钟。向每种样品中加入两倍体积的2%多聚甲醛,并在室温下孵育10分钟。然后将细胞用1ml冰冷的PBS洗涤一次,重悬于200μl PBS中,并使用FACScalibur流式细胞仪分析。使用每个样品2x10 4个事件的几何平均荧光强度(GMFI)来计算内化抗体的百分比。由于在4℃不会发生内化,所以在冰上孵育随后与抗-Alexa488抗体孵育的样品中测量的荧光被认为是不可淬灭的背景表面荧光,并在淬灭前从37℃孵育的样品所得GMFI值中扣除。内化如下计算:内化百分比=(GMFI淬灭/GMFI未淬灭)*100。
结果在图8中示出。双特异性抗体SI-4X2(39.3%)比单特异性对照抗体SI-4C1(18.29%)和SI-4C2(14.97%)以及SI-4C1+SI-4C2的组合(29.19%)更大程度地内化。
药物组合物
词语“有效量”是指有效实现期望效果的药物的量,例如改善患者的疾病。其中该疾病是癌症,有效量的药物可以抑制(例如,在一定程度上减缓、抑制或停止)以下一个或多个示例性特征,包括但不限于癌细胞生长、癌细胞增殖、癌细胞运动、癌细胞浸润到周围器官、肿瘤转移以及肿瘤生长。其中该疾病是癌症,当对患者施药时,有效量的药物可以可选地进行以下一种或多种:减缓或停止肿瘤生长、减小肿瘤尺寸(例如体积或质量)、在一定程度上缓解一种或多种与癌相关的症状、延长无进展生存期、产生客观疗效(包括例如部分疗效或完全疗效),以及增加卵巢生存时间。在药物可以预防生长和/或杀死现有癌细胞的程度上,其是抑制细胞生长和/或细胞毒性的。
关于给诸如需要治疗的人类患者的患者施用合适的组合物的制剂,本文公开的抗体可以根据所选择的施药途径与本领域已知的药学上可接受的载体混合或组合。对本文公开的抗体的应用模式没有特别限制,并且合适的施药途径和合适的组合物的选择在本领域中是已知的而无需过多的实验。
虽然许多施药形式是可能的,但是示例性施药形式将是注射溶液,具体用于静脉内或动脉内注射。通常,用于注射的合适的药物组合物可以包括药学上合适的载体或赋形剂,诸如但不限于缓冲液、表面活性剂或稳定剂。示例性缓冲液可以包括但不限于乙酸盐,磷酸盐或柠檬酸盐缓冲液。示例性表面活性剂可以包括但不限于聚山梨醇酯。示例性稳定剂可以包括但不限于人白蛋白。
类似地,本领域技术人员有能力确定其中公开的抗体的有效量或浓度以有效治疗诸如癌症的病症。其他参数,诸如药物组合物中各种组分的比例、施药剂量以及频率可由本领域技术人员获得,而无需过多的实验。例如,合适的注射溶液可以包含但不限于约1至约20,约1至约10mg抗体/ml。示例性剂量可以是但不限于约0.1至约20,约1至约5mg/Kg体重。示例性施药频率可以是但不限于每天一次或每周三次。
虽然已经参考特定实施例或示例描述了本公开,但是可以理解,实施例是说明性的,并且公开范围不受此限制。本公开的可选实施例对于本公开所属领域的普通技术人员来说可以变得显而易见。这些可选实施例被认为包含在本公开的范围内。因此,本公开的范围由所附权利要求限定,并且由前述描述支持。
Claims (10)
1.一种双特异性四价抗体,所述双特异性四价抗体包含:
两条IgGl重链;
两条kappa轻链;以及
两个单链Fv(scFv)结构域。
其中所述两条IgGl重链和kappa轻链形成对HER2的第一结构域具有结合特异性的IgG部分;
其中所述两个scFv结构域对HER2的第二结构域具有结合特异性,并且每个scFv结构域通过具有氨基酸序列(gly-gly-gly-gly-ser)n((G4S)n)的连接体与任一个IgGl重链的C端残基连接;其中n是至少为2-20的整数;并且
其中每个scFv结构域具有N端-可变重链结构域-接头-可变轻链结构域-C端或N端-可变轻链结构域-接头-可变重链结构域-C端的结构顺序,并且其中所述接头可以由(gly-gly-gly-gly-ser)m((G4S)m)氨基酸序列组成。其中m是至少为3-30的整数。
2.根据权利要求1所述的双特异性四价抗体,其中,
所述IgGl重链中的至少一个是人源化的或人IgGl重链。
3.根据权利要求1所述的双特异性四价抗体,其中,
两条IgGl重链是人源化的或人IgGl重链。
4.根据权利要求1所述的双特异性四价抗体,其中,
所述kappa轻链中的至少一个是人源化的或人kappa轻链。
5.根据权利要求1所述的双特异性四价抗体,其中,
两条kappa轻链是人源化的或人kappa轻链。
6.根据权利要求1所述的双特异性四价抗体,其中,
可变轻链中的至少一个包含选自SEQ ID NO 36的氨基酸序列。
7.根据权利要求1所述的双特异性四价抗体,其中,
可变重链中的至少一个包含选自SEQ ID NO 41的氨基酸序列。
8.根据权利要求1所述的双特异性四价抗体,其中,scFv结构域中的至少一个包含选自SEQ ID NO 42的氨基酸序列。
9.根据权利要求1所述的双特异性四价抗体,其中,
所述IgGl重链中的至少一个包含SEQ ID NO40的氨基酸序列。
10.根据权利要求1所述的双特异性四价抗体,其中,
所述IgG部分对HER2的结构域4具有结合特异性,并且所述scFv结构域对HER2的结构域2具有结合特异性;其中所述IgGl重链、连接体以及scFv结构域具有SEQ ID NO 40的氨基酸序列,并且所述kappa轻链具有SEQ ID NO 35的氨基酸序列。
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201462095348P | 2014-12-22 | 2014-12-22 | |
US62/095,348 | 2014-12-22 | ||
CN201580073430.9A CN107206074B (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制备和使用方法 |
PCT/US2015/066952 WO2016106158A1 (en) | 2014-12-22 | 2015-12-19 | Bispecific tetravalent antibodies and methods of making and using thereof |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201580073430.9A Division CN107206074B (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制备和使用方法 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN117467017A true CN117467017A (zh) | 2024-01-30 |
Family
ID=56151464
Family Applications (11)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110470781.0A Pending CN113105552A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN201580036408.7A Active CN106659779B (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN201580073430.9A Active CN107206074B (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制备和使用方法 |
CN202110471245.2A Pending CN113512123A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN202110470743.5A Pending CN113105551A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN202110471082.8A Pending CN113512122A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN202110471052.7A Pending CN113512121A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN202110471476.3A Pending CN113150165A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN202110470818.XA Pending CN113512120A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN202110471392.XA Pending CN113105553A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN202311507433.1A Pending CN117467017A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制备和使用方法 |
Family Applications Before (10)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110470781.0A Pending CN113105552A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN201580036408.7A Active CN106659779B (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN201580073430.9A Active CN107206074B (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制备和使用方法 |
CN202110471245.2A Pending CN113512123A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN202110470743.5A Pending CN113105551A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN202110471082.8A Pending CN113512122A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN202110471052.7A Pending CN113512121A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN202110471476.3A Pending CN113150165A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN202110470818.XA Pending CN113512120A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
CN202110471392.XA Pending CN113105553A (zh) | 2014-12-22 | 2015-12-19 | 双特异性四价抗体及其制造和使用方法 |
Country Status (12)
Country | Link |
---|---|
US (2) | US10919977B2 (zh) |
EP (2) | EP3237005A4 (zh) |
JP (2) | JP6947639B2 (zh) |
KR (3) | KR20230149328A (zh) |
CN (11) | CN113105552A (zh) |
AU (1) | AU2015369831B2 (zh) |
CA (2) | CA3138083A1 (zh) |
ES (1) | ES2962675T3 (zh) |
IL (3) | IL286835B2 (zh) |
NZ (1) | NZ732628A (zh) |
SG (1) | SG11201704741PA (zh) |
WO (2) | WO2016106158A1 (zh) |
Families Citing this family (24)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA3138083A1 (en) * | 2014-12-22 | 2016-06-30 | Systimmune, Inc. | Bispecific tetravalent antibodies and methods of making and using thereof |
CN106632681B (zh) * | 2016-10-11 | 2017-11-14 | 北京东方百泰生物科技有限公司 | 抗egfr和抗cd3双特异抗体及其应用 |
CN108948195B (zh) * | 2017-05-23 | 2022-05-31 | 胡毅 | 一种抗egfr/pd-l1双靶向抗体、其制备方法及用途 |
CN116769035A (zh) * | 2017-06-25 | 2023-09-19 | 西雅图免疫公司 | 抗4-1bb抗体及其制备和使用方法 |
AU2018358138C1 (en) * | 2017-11-02 | 2022-12-08 | Baili-Bio (Chengdu) Pharmaceutical Co., Ltd. | Bispecific antibodies and methods of making and using thereof |
KR20200109313A (ko) * | 2018-01-15 | 2020-09-22 | 난징 레전드 바이오테크 씨오., 엘티디. | Tigit에 대한 항체 및 이의 변이체 |
CN108220244A (zh) * | 2018-01-18 | 2018-06-29 | 东北农业大学 | 一种含重组人表皮生长因子受体抗体基因的cho细胞株、筛选方法及其生产工艺 |
CN107974461A (zh) * | 2018-01-18 | 2018-05-01 | 东北农业大学 | 一种高效表达anti-hEGFR基因的真核表达载体构建方法 |
CN113817066A (zh) * | 2018-04-13 | 2021-12-21 | 艾菲默德有限责任公司 | 自然杀伤细胞接合抗体融合构建体 |
US20210115152A1 (en) * | 2018-05-16 | 2021-04-22 | Arbele Limited | Composition of bispecific antibodies and method of use thereof |
EP3833690B1 (en) * | 2018-08-08 | 2024-03-27 | ImmuneOnco Biopharmaceuticals (Shanghai) Inc. | Recombinant bifunctional protein targeting cd47 and her2 |
CN110850068B (zh) * | 2018-08-21 | 2023-08-15 | 上海恒润达生生物科技股份有限公司 | 一种嵌合抗原受体亲和力检测方法 |
EP3880247A4 (en) * | 2018-11-13 | 2022-10-26 | JN Biosciences, LLC | BISPECIFIC ANTIBODIES TO ACTIVATE IMMUNE CELLS |
AU2020327000A1 (en) | 2019-08-08 | 2022-03-31 | Regeneron Pharmaceuticals, Inc. | Novel antigen binding molecule formats |
TW202200619A (zh) * | 2020-03-17 | 2022-01-01 | 美商西雅圖免疫公司 | 引導及導航控制(gnc)抗體樣蛋白質及製造與使用其之方法 |
CN111548424A (zh) * | 2020-06-05 | 2020-08-18 | 上海科弈药业科技有限公司 | 一种靶向egfr与cd47的多功能融合蛋白及其应用 |
IL301473A (en) * | 2020-09-21 | 2023-05-01 | Systimmune Inc | EGFR binding complex and how it is created and used |
CN117940459A (zh) * | 2021-08-25 | 2024-04-26 | 西雅图免疫公司 | 靶向egfr和her3的双特异性四价抗体 |
WO2023044483A2 (en) * | 2021-09-20 | 2023-03-23 | Voyager Therapeutics, Inc. | Compositions and methods for the treatment of her2 positive cancer |
AU2022357570A1 (en) * | 2021-10-03 | 2024-04-11 | Systimmune, Inc. | Methods of treating cancer and the pharmaceutical compositions thereof |
CA3237844A1 (en) * | 2021-11-15 | 2023-05-19 | Systimmune, Inc. | Bispecific antibody-camptothecin drug conjugate and pharmaceutical use thereof |
WO2023241480A1 (zh) * | 2022-06-13 | 2023-12-21 | 三优生物医药(上海)有限公司 | 抗pd-l1、vegf和egfr三特异性抗体及其应用 |
WO2024050439A2 (en) * | 2022-08-31 | 2024-03-07 | Systimmune, Inc. | Biepitopic tetravalent antibody targeting egfr |
CN116063564A (zh) * | 2022-10-08 | 2023-05-05 | 盛禾(中国)生物制药有限公司 | 一种纯化融合蛋白的方法 |
Family Cites Families (30)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB2200753B (en) | 1986-12-23 | 1991-01-23 | Brookes & Gatehouse | Speed measurement device |
US5641870A (en) | 1995-04-20 | 1997-06-24 | Genentech, Inc. | Low pH hydrophobic interaction chromatography for antibody purification |
WO2003066662A2 (en) * | 2002-02-05 | 2003-08-14 | Genentech, Inc. | Protein purification |
US7332585B2 (en) | 2002-04-05 | 2008-02-19 | The Regents Of The California University | Bispecific single chain Fv antibody molecules and methods of use thereof |
UA94213C2 (ru) * | 2004-09-17 | 2011-04-26 | Домантис Лимитед | Применение полипептида одноцепочечного антитела, который подавляет активность cd40 или cd40l в приготовлении медикамента для лечения аутоиммунного заболевания |
WO2007147001A2 (en) * | 2006-06-14 | 2007-12-21 | Imclone Systems Incorporated | Lyophilized formulations of anti-egfr antibodies |
AU2007353412A1 (en) * | 2006-11-21 | 2008-11-20 | Fox Chase Cancer Center | Anti-EGFR family antibodies, bispecific anti-EGFR family antibodies and methods of use thereof |
AR075896A1 (es) * | 2009-03-20 | 2011-05-04 | Genentech Inc | Anticuerpos anti-her (factor de crecimiento epidermico) |
KR20110134494A (ko) | 2009-03-27 | 2011-12-14 | 자이모제네틱스, 인코포레이티드 | 항체-수용체 조합물을 포함하는 다중특이적-결합 단백질을 사용하기 위한 조성물 및 방법 |
CA2756244A1 (en) * | 2009-04-02 | 2010-10-07 | Roche Glycart Ag | Multispecific antibodies comprising full length antibodies and single chain fab fragments |
MX2011010166A (es) * | 2009-04-07 | 2011-10-11 | Roche Glycart Ag | Anticuerpos biespecificos anti-erbb-3/anti-c-met. |
EP2473524A4 (en) * | 2009-09-01 | 2013-05-22 | Abbott Lab | IMMUNOGLOBULINE WITH DOUBLE VARIABLE DOMAIN AND ITS USE |
EP2496598B1 (en) * | 2009-11-04 | 2017-08-02 | Affibody AB | Her3 binding polypeptides |
CA2811747A1 (en) * | 2010-09-30 | 2012-04-05 | Merck Sharp & Dohme Corp. | Generation, characterization and uses thereof of anti-her3 antibodies |
CN103384681B (zh) * | 2010-12-23 | 2018-05-18 | 霍夫曼-拉罗奇有限公司 | 结合剂 |
TWI803876B (zh) | 2011-03-28 | 2023-06-01 | 法商賽諾菲公司 | 具有交叉結合區定向之雙重可變區類抗體結合蛋白 |
US20140170148A1 (en) * | 2011-04-20 | 2014-06-19 | Genmab A/S | Bispecific antibodies against her2 |
CA2832387A1 (en) * | 2011-04-20 | 2012-10-26 | Genmab A/S | Bispecifc antibodies against her2 |
PL2703486T3 (pl) * | 2011-04-25 | 2018-07-31 | Daiichi Sankyo Company, Limited | Przeciwciało anty-b7-h3 |
CN102250246A (zh) * | 2011-06-10 | 2011-11-23 | 常州亚当生物技术有限公司 | 抗VEGF/PDGFRβ双特异性抗体及其应用 |
CN102250247B (zh) | 2011-06-15 | 2013-06-19 | 常州亚当生物技术有限公司 | 一种抗vegf/ang2双特异性抗体及其应用 |
CN104203981A (zh) * | 2011-12-19 | 2014-12-10 | 合成免疫股份有限公司 | 双特异性抗体分子 |
CN104144949B (zh) * | 2011-12-22 | 2016-08-31 | 财团法人生物技术开发中心 | 双特异性t细胞活化剂抗体 |
WO2013148315A1 (en) * | 2012-03-27 | 2013-10-03 | Genentech, Inc. | Diagnosis and treatments relating to her3 inhibitors |
IN2015DN01115A (zh) * | 2012-07-13 | 2015-06-26 | Zymeworks Inc | |
SI2922872T1 (sl) * | 2012-11-21 | 2019-01-31 | Janssen Biotech, Inc., | Bispecifična protitelesa EGFR/C-MET |
CN103333179B (zh) * | 2012-12-21 | 2017-06-16 | 百奥泰生物科技(广州)有限公司 | 类美登素衍生物及其制备方法和用途 |
CN105189557A (zh) | 2013-03-15 | 2015-12-23 | 默克专利有限公司 | 四价双特异性抗体 |
CN104211814A (zh) * | 2013-05-29 | 2014-12-17 | 三星电子株式会社 | 用于消耗靶膜蛋白的组合物 |
CA3138083A1 (en) | 2014-12-22 | 2016-06-30 | Systimmune, Inc. | Bispecific tetravalent antibodies and methods of making and using thereof |
-
2015
- 2015-12-19 CA CA3138083A patent/CA3138083A1/en active Pending
- 2015-12-19 EP EP15874216.3A patent/EP3237005A4/en active Pending
- 2015-12-19 CN CN202110470781.0A patent/CN113105552A/zh active Pending
- 2015-12-19 CN CN201580036408.7A patent/CN106659779B/zh active Active
- 2015-12-19 CN CN201580073430.9A patent/CN107206074B/zh active Active
- 2015-12-19 KR KR1020237034930A patent/KR20230149328A/ko not_active Application Discontinuation
- 2015-12-19 US US15/119,694 patent/US10919977B2/en active Active
- 2015-12-19 ES ES15874217T patent/ES2962675T3/es active Active
- 2015-12-19 CN CN202110471245.2A patent/CN113512123A/zh active Pending
- 2015-12-19 AU AU2015369831A patent/AU2015369831B2/en active Active
- 2015-12-19 WO PCT/US2015/066952 patent/WO2016106158A1/en active Application Filing
- 2015-12-19 CN CN202110470743.5A patent/CN113105551A/zh active Pending
- 2015-12-19 CN CN202110471082.8A patent/CN113512122A/zh active Pending
- 2015-12-19 EP EP15874217.1A patent/EP3237006B1/en active Active
- 2015-12-19 US US15/538,189 patent/US10717783B2/en active Active
- 2015-12-19 KR KR1020237021346A patent/KR102590385B1/ko active IP Right Grant
- 2015-12-19 WO PCT/US2015/066951 patent/WO2016106157A1/en active Application Filing
- 2015-12-19 CN CN202110471052.7A patent/CN113512121A/zh active Pending
- 2015-12-19 CN CN202110471476.3A patent/CN113150165A/zh active Pending
- 2015-12-19 CN CN202110470818.XA patent/CN113512120A/zh active Pending
- 2015-12-19 KR KR1020177020104A patent/KR102548827B1/ko active IP Right Grant
- 2015-12-19 IL IL286835A patent/IL286835B2/en unknown
- 2015-12-19 CN CN202110471392.XA patent/CN113105553A/zh active Pending
- 2015-12-19 IL IL305193A patent/IL305193A/en unknown
- 2015-12-19 SG SG11201704741PA patent/SG11201704741PA/en unknown
- 2015-12-19 CA CA2969867A patent/CA2969867C/en active Active
- 2015-12-19 JP JP2017551580A patent/JP6947639B2/ja active Active
- 2015-12-19 CN CN202311507433.1A patent/CN117467017A/zh active Pending
- 2015-12-19 NZ NZ732628A patent/NZ732628A/en unknown
-
2017
- 2017-06-11 IL IL252811A patent/IL252811B/en unknown
-
2021
- 2021-04-15 JP JP2021068859A patent/JP7335290B2/ja active Active
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US10717783B2 (en) | Bispecific tetravalent antibodies and methods of making and using thereof | |
JP7165265B2 (ja) | Her2/pd1二重特異性抗体 | |
CN116199785A (zh) | 多特异性抗体及其制备和使用方法 | |
CN110799540A (zh) | 多特异性抗体及其制备和使用方法 | |
CN112111008B (zh) | 抗cd73抗体及其应用 | |
US11680102B2 (en) | Anti-BAFF receptor antibodies and uses thereof | |
WO2023098770A1 (zh) | 抗trop-2/pd-l1双特异性抗体 | |
WO2021244553A1 (zh) | 一种抗pd-l1和egfr的四价双特异性抗体 | |
US20210277128A1 (en) | Bispecific Tetravalent Antibodies and Methods of Making and Using Thereof | |
WO2022111535A1 (zh) | 结合her2的多价双特异性抗体、其制备方法和用途 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |