WO2015095972A1 - Antibodies comprising c-terminal light chain polypeptide extensions and conjugates and methods of use thereof - Google Patents
Antibodies comprising c-terminal light chain polypeptide extensions and conjugates and methods of use thereof Download PDFInfo
- Publication number
- WO2015095972A1 WO2015095972A1 PCT/CA2014/051263 CA2014051263W WO2015095972A1 WO 2015095972 A1 WO2015095972 A1 WO 2015095972A1 CA 2014051263 W CA2014051263 W CA 2014051263W WO 2015095972 A1 WO2015095972 A1 WO 2015095972A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- antibody
- amino acid
- extension
- agent
- light chain
- Prior art date
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 204
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 202
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 202
- 238000000034 method Methods 0.000 title claims abstract description 83
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 148
- 125000001433 C-terminal amino-acid group Chemical group 0.000 claims abstract description 127
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 claims abstract description 125
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 69
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 64
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 64
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 37
- 229940127121 immunoconjugate Drugs 0.000 claims abstract description 34
- 210000004027 cell Anatomy 0.000 claims description 136
- 235000018417 cysteine Nutrition 0.000 claims description 105
- 125000006850 spacer group Chemical group 0.000 claims description 102
- 239000000427 antigen Substances 0.000 claims description 89
- 108091007433 antigens Proteins 0.000 claims description 88
- 102000036639 antigens Human genes 0.000 claims description 88
- 150000001413 amino acids Chemical class 0.000 claims description 78
- 235000001014 amino acid Nutrition 0.000 claims description 72
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims description 63
- 239000003814 drug Substances 0.000 claims description 60
- 230000027455 binding Effects 0.000 claims description 49
- 150000001945 cysteines Chemical class 0.000 claims description 42
- 125000003396 thiol group Chemical group [H]S* 0.000 claims description 33
- 238000006243 chemical reaction Methods 0.000 claims description 30
- 239000012634 fragment Substances 0.000 claims description 29
- 238000002372 labelling Methods 0.000 claims description 26
- 230000002829 reductive effect Effects 0.000 claims description 25
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 24
- 239000013598 vector Substances 0.000 claims description 21
- 229940124597 therapeutic agent Drugs 0.000 claims description 19
- 239000004471 Glycine Substances 0.000 claims description 12
- 230000001268 conjugating effect Effects 0.000 claims description 12
- PEEHTFAAVSWFBL-UHFFFAOYSA-N Maleimide Chemical compound O=C1NC(=O)C=C1 PEEHTFAAVSWFBL-UHFFFAOYSA-N 0.000 claims description 11
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims description 11
- 229940127089 cytotoxic agent Drugs 0.000 claims description 11
- 239000002254 cytotoxic agent Substances 0.000 claims description 11
- 102000016266 T-Cell Antigen Receptors Human genes 0.000 claims description 10
- 238000011503 in vivo imaging Methods 0.000 claims description 10
- 231100000599 cytotoxic agent Toxicity 0.000 claims description 9
- 108091008874 T cell receptors Proteins 0.000 claims description 8
- 239000012216 imaging agent Substances 0.000 claims description 8
- 125000005179 haloacetyl group Chemical group 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 claims description 7
- HAXFWIACAGNFHA-UHFFFAOYSA-N aldrithiol Chemical compound C=1C=CC=NC=1SSC1=CC=CC=N1 HAXFWIACAGNFHA-UHFFFAOYSA-N 0.000 claims description 5
- AFOSIXZFDONLBT-UHFFFAOYSA-N divinyl sulfone Chemical compound C=CS(=O)(=O)C=C AFOSIXZFDONLBT-UHFFFAOYSA-N 0.000 claims description 5
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 5
- 238000004132 cross linking Methods 0.000 claims description 4
- 210000001236 prokaryotic cell Anatomy 0.000 claims description 4
- 210000003527 eukaryotic cell Anatomy 0.000 claims 1
- 210000004899 c-terminal region Anatomy 0.000 abstract description 116
- 239000000562 conjugate Substances 0.000 description 102
- 239000003053 toxin Substances 0.000 description 65
- 229940024606 amino acid Drugs 0.000 description 58
- 125000003275 alpha amino acid group Chemical group 0.000 description 57
- 229960002433 cysteine Drugs 0.000 description 53
- 230000021615 conjugation Effects 0.000 description 46
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 41
- 229940079593 drug Drugs 0.000 description 40
- 229940022353 herceptin Drugs 0.000 description 40
- 206010028980 Neoplasm Diseases 0.000 description 33
- 239000000611 antibody drug conjugate Substances 0.000 description 33
- 229940049595 antibody-drug conjugate Drugs 0.000 description 33
- 238000000338 in vitro Methods 0.000 description 30
- 108090000623 proteins and genes Proteins 0.000 description 30
- 239000000203 mixture Substances 0.000 description 29
- 238000004191 hydrophobic interaction chromatography Methods 0.000 description 28
- 102000004169 proteins and genes Human genes 0.000 description 28
- 235000018102 proteins Nutrition 0.000 description 27
- 238000011068 loading method Methods 0.000 description 25
- -1 cysteine amino acids Chemical class 0.000 description 24
- 238000006722 reduction reaction Methods 0.000 description 24
- 238000001516 cell proliferation assay Methods 0.000 description 22
- 239000007795 chemical reaction product Substances 0.000 description 22
- 230000009467 reduction Effects 0.000 description 21
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 20
- 239000000539 dimer Substances 0.000 description 19
- 239000000872 buffer Substances 0.000 description 17
- 238000009826 distribution Methods 0.000 description 17
- 201000011510 cancer Diseases 0.000 description 15
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 description 14
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 description 14
- 239000003638 chemical reducing agent Substances 0.000 description 14
- 241000894007 species Species 0.000 description 14
- 231100000765 toxin Toxicity 0.000 description 14
- 108700012359 toxins Proteins 0.000 description 14
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 12
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 12
- 230000037396 body weight Effects 0.000 description 12
- 201000010099 disease Diseases 0.000 description 12
- 238000009472 formulation Methods 0.000 description 12
- 238000003752 polymerase chain reaction Methods 0.000 description 12
- 229960000575 trastuzumab Drugs 0.000 description 12
- 108060003951 Immunoglobulin Proteins 0.000 description 11
- 150000001875 compounds Chemical class 0.000 description 11
- 102000018358 immunoglobulin Human genes 0.000 description 11
- 101000874088 Centruroides noxius Toxin Cn3 Proteins 0.000 description 10
- 101000761023 Loxosceles intermedia U2-sicaritoxin-Li1a Proteins 0.000 description 10
- 101000675481 Naja sputatrix Neurotoxin 3 Proteins 0.000 description 10
- 101000723129 Oxyuranus scutellatus scutellatus Short neurotoxin 1 Proteins 0.000 description 10
- 238000010367 cloning Methods 0.000 description 10
- 239000013604 expression vector Substances 0.000 description 10
- 239000000243 solution Substances 0.000 description 10
- 210000001519 tissue Anatomy 0.000 description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 9
- 238000004458 analytical method Methods 0.000 description 9
- 230000000694 effects Effects 0.000 description 9
- 239000000499 gel Substances 0.000 description 9
- 238000011534 incubation Methods 0.000 description 9
- 239000007924 injection Substances 0.000 description 9
- 238000002347 injection Methods 0.000 description 9
- 238000011282 treatment Methods 0.000 description 9
- 230000015572 biosynthetic process Effects 0.000 description 8
- 208000035475 disorder Diseases 0.000 description 8
- 230000006870 function Effects 0.000 description 8
- 239000003550 marker Substances 0.000 description 8
- 230000009870 specific binding Effects 0.000 description 8
- 238000007792 addition Methods 0.000 description 7
- 238000006467 substitution reaction Methods 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 108010044023 Ki-1 Antigen Proteins 0.000 description 6
- 238000003556 assay Methods 0.000 description 6
- 239000002552 dosage form Substances 0.000 description 6
- 238000003384 imaging method Methods 0.000 description 6
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 238000000746 purification Methods 0.000 description 6
- 239000004094 surface-active agent Substances 0.000 description 6
- 208000024891 symptom Diseases 0.000 description 6
- 150000003573 thiols Chemical class 0.000 description 6
- 238000001890 transfection Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 5
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 5
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 5
- 108091028043 Nucleic acid sequence Proteins 0.000 description 5
- 241000235648 Pichia Species 0.000 description 5
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 5
- 230000009471 action Effects 0.000 description 5
- 238000013459 approach Methods 0.000 description 5
- UFULAYFCSOUIOV-UHFFFAOYSA-N cysteamine Chemical compound NCCS UFULAYFCSOUIOV-UHFFFAOYSA-N 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 5
- 229940072221 immunoglobulins Drugs 0.000 description 5
- 238000001727 in vivo Methods 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 230000003211 malignant effect Effects 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 229960003151 mercaptamine Drugs 0.000 description 5
- QPCDCPDFJACHGM-UHFFFAOYSA-K pentetate(3-) Chemical compound OC(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O QPCDCPDFJACHGM-UHFFFAOYSA-K 0.000 description 5
- 239000013612 plasmid Substances 0.000 description 5
- 229920001223 polyethylene glycol Polymers 0.000 description 5
- 239000003755 preservative agent Substances 0.000 description 5
- 235000002639 sodium chloride Nutrition 0.000 description 5
- 239000007790 solid phase Substances 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 4
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 4
- 101001023095 Anemonia sulcata Delta-actitoxin-Avd1a Proteins 0.000 description 4
- 101000641989 Araneus ventricosus Kunitz-type U1-aranetoxin-Av1a Proteins 0.000 description 4
- 101001028691 Carybdea rastonii Toxin CrTX-A Proteins 0.000 description 4
- 101000685083 Centruroides infamatus Beta-toxin Cii1 Proteins 0.000 description 4
- 101000685085 Centruroides noxius Toxin Cn1 Proteins 0.000 description 4
- 101001028688 Chironex fleckeri Toxin CfTX-1 Proteins 0.000 description 4
- 101000644407 Cyriopagopus schmidti U6-theraphotoxin-Hs1a Proteins 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 4
- SBJKKFFYIZUCET-JLAZNSOCSA-N Dehydro-L-ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(=O)C1=O SBJKKFFYIZUCET-JLAZNSOCSA-N 0.000 description 4
- SBJKKFFYIZUCET-UHFFFAOYSA-N Dehydroascorbic acid Natural products OCC(O)C1OC(=O)C(=O)C1=O SBJKKFFYIZUCET-UHFFFAOYSA-N 0.000 description 4
- 101150029707 ERBB2 gene Proteins 0.000 description 4
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 229910019142 PO4 Inorganic materials 0.000 description 4
- 101000679608 Phaeosphaeria nodorum (strain SN15 / ATCC MYA-4574 / FGSC 10173) Cysteine rich necrotrophic effector Tox1 Proteins 0.000 description 4
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 4
- 239000002202 Polyethylene glycol Substances 0.000 description 4
- 108010034634 Repressor Proteins Proteins 0.000 description 4
- 102000009661 Repressor Proteins Human genes 0.000 description 4
- 241000283984 Rodentia Species 0.000 description 4
- 101710084578 Short neurotoxin 1 Proteins 0.000 description 4
- PZBFGYYEXUXCOF-UHFFFAOYSA-N TCEP Chemical compound OC(=O)CCP(CCC(O)=O)CCC(O)=O PZBFGYYEXUXCOF-UHFFFAOYSA-N 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 4
- 238000013270 controlled release Methods 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- 235000020960 dehydroascorbic acid Nutrition 0.000 description 4
- 239000011615 dehydroascorbic acid Substances 0.000 description 4
- 238000000113 differential scanning calorimetry Methods 0.000 description 4
- 229960002989 glutamic acid Drugs 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 239000008101 lactose Substances 0.000 description 4
- RLSSMJSEOOYNOY-UHFFFAOYSA-N m-cresol Chemical compound CC1=CC=CC(O)=C1 RLSSMJSEOOYNOY-UHFFFAOYSA-N 0.000 description 4
- 210000004962 mammalian cell Anatomy 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 235000006109 methionine Nutrition 0.000 description 4
- 239000002773 nucleotide Substances 0.000 description 4
- 230000036961 partial effect Effects 0.000 description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 4
- 239000010452 phosphate Substances 0.000 description 4
- 229920001983 poloxamer Polymers 0.000 description 4
- 229920000136 polysorbate Polymers 0.000 description 4
- 239000011541 reaction mixture Substances 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 239000003381 stabilizer Substances 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- 238000013519 translation Methods 0.000 description 4
- 150000003923 2,5-pyrrolediones Chemical class 0.000 description 3
- OGMADIBCHLQMIP-UHFFFAOYSA-N 2-aminoethanethiol;hydron;chloride Chemical group Cl.NCCS OGMADIBCHLQMIP-UHFFFAOYSA-N 0.000 description 3
- 229920002261 Corn starch Polymers 0.000 description 3
- 102000001301 EGF receptor Human genes 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 108050001049 Extracellular proteins Proteins 0.000 description 3
- 206010056740 Genital discharge Diseases 0.000 description 3
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 3
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 3
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 3
- 229930126263 Maytansine Natural products 0.000 description 3
- 101100018611 Mus musculus Igkc gene Proteins 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 241000589516 Pseudomonas Species 0.000 description 3
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 239000000654 additive Substances 0.000 description 3
- 235000004279 alanine Nutrition 0.000 description 3
- 229960000455 brentuximab vedotin Drugs 0.000 description 3
- 230000004663 cell proliferation Effects 0.000 description 3
- 230000003833 cell viability Effects 0.000 description 3
- 230000001413 cellular effect Effects 0.000 description 3
- 239000008120 corn starch Substances 0.000 description 3
- 229940097265 cysteamine hydrochloride Drugs 0.000 description 3
- 238000007405 data analysis Methods 0.000 description 3
- 238000013480 data collection Methods 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 239000000975 dye Substances 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- 235000013922 glutamic acid Nutrition 0.000 description 3
- 239000004220 glutamic acid Substances 0.000 description 3
- 235000014304 histidine Nutrition 0.000 description 3
- 239000003112 inhibitor Substances 0.000 description 3
- 238000003780 insertion Methods 0.000 description 3
- 230000037431 insertion Effects 0.000 description 3
- 230000000670 limiting effect Effects 0.000 description 3
- WKPWGQKGSOKKOO-RSFHAFMBSA-N maytansine Chemical compound CO[C@@H]([C@@]1(O)C[C@](OC(=O)N1)([C@H]([C@@H]1O[C@@]1(C)[C@@H](OC(=O)[C@H](C)N(C)C(C)=O)CC(=O)N1C)C)[H])\C=C\C=C(C)\CC2=CC(OC)=C(Cl)C1=C2 WKPWGQKGSOKKOO-RSFHAFMBSA-N 0.000 description 3
- 229910052751 metal Inorganic materials 0.000 description 3
- 239000002184 metal Substances 0.000 description 3
- 229930182817 methionine Natural products 0.000 description 3
- 229960004452 methionine Drugs 0.000 description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 125000003729 nucleotide group Chemical group 0.000 description 3
- 229920001993 poloxamer 188 Polymers 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 235000000346 sugar Nutrition 0.000 description 3
- 150000008163 sugars Chemical class 0.000 description 3
- 239000003826 tablet Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 150000003568 thioethers Chemical class 0.000 description 3
- 230000014616 translation Effects 0.000 description 3
- 235000002374 tyrosine Nutrition 0.000 description 3
- 238000004704 ultra performance liquid chromatography Methods 0.000 description 3
- 210000005253 yeast cell Anatomy 0.000 description 3
- KQODQNJLJQHFQV-MKWZWQCGSA-N (e,4s)-4-[[(2s)-3,3-dimethyl-2-[[(2s)-3-methyl-2-(methylamino)-3-(1-methylindol-3-yl)butanoyl]amino]butanoyl]-methylamino]-2,5-dimethylhex-2-enoic acid Chemical compound C1=CC=C2C(C(C)(C)[C@@H](C(=O)N[C@H](C(=O)N(C)[C@H](\C=C(/C)C(O)=O)C(C)C)C(C)(C)C)NC)=CN(C)C2=C1 KQODQNJLJQHFQV-MKWZWQCGSA-N 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 description 2
- VPFUWHKTPYPNGT-UHFFFAOYSA-N 3-(3,4-dihydroxyphenyl)-1-(5-hydroxy-2,2-dimethylchromen-6-yl)propan-1-one Chemical compound OC1=C2C=CC(C)(C)OC2=CC=C1C(=O)CCC1=CC=C(O)C(O)=C1 VPFUWHKTPYPNGT-UHFFFAOYSA-N 0.000 description 2
- PBVAJRFEEOIAGW-UHFFFAOYSA-N 3-[bis(2-carboxyethyl)phosphanyl]propanoic acid;hydrochloride Chemical compound Cl.OC(=O)CCP(CCC(O)=O)CCC(O)=O PBVAJRFEEOIAGW-UHFFFAOYSA-N 0.000 description 2
- CFKMVGJGLGKFKI-UHFFFAOYSA-N 4-chloro-m-cresol Chemical compound CC1=CC(O)=CC=C1Cl CFKMVGJGLGKFKI-UHFFFAOYSA-N 0.000 description 2
- 108010066676 Abrin Proteins 0.000 description 2
- 101000939689 Araneus ventricosus U2-aranetoxin-Av1a Proteins 0.000 description 2
- 239000004475 Arginine Substances 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 244000063299 Bacillus subtilis Species 0.000 description 2
- 235000014469 Bacillus subtilis Nutrition 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 206010006187 Breast cancer Diseases 0.000 description 2
- 208000026310 Breast neoplasm Diseases 0.000 description 2
- 101000633673 Buthacus arenicola Beta-insect depressant toxin BaIT2 Proteins 0.000 description 2
- 101000654318 Centruroides noxius Beta-mammal toxin Cn2 Proteins 0.000 description 2
- 101001028695 Chironex fleckeri Toxin CfTX-2 Proteins 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- 108010053187 Diphtheria Toxin Proteins 0.000 description 2
- 102000016607 Diphtheria Toxin Human genes 0.000 description 2
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 2
- 108060006698 EGF receptor Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- 108700004714 Gelonium multiflorum GEL Proteins 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 102100041003 Glutamate carboxypeptidase 2 Human genes 0.000 description 2
- 102000009465 Growth Factor Receptors Human genes 0.000 description 2
- 108010009202 Growth Factor Receptors Proteins 0.000 description 2
- 101000892862 Homo sapiens Glutamate carboxypeptidase 2 Proteins 0.000 description 2
- 101000896379 Homo sapiens Transmembrane reductase CYB561D2 Proteins 0.000 description 2
- 108090000144 Human Proteins Proteins 0.000 description 2
- 102000003839 Human Proteins Human genes 0.000 description 2
- 241000725303 Human immunodeficiency virus Species 0.000 description 2
- 108010058683 Immobilized Proteins Proteins 0.000 description 2
- 108700005091 Immunoglobulin Genes Proteins 0.000 description 2
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 2
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 2
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 2
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 2
- 125000002707 L-tryptophyl group Chemical group [H]C1=C([H])C([H])=C2C(C([C@](N([H])[H])(C(=O)[*])[H])([H])[H])=C([H])N([H])C2=C1[H] 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- 229930195725 Mannitol Natural products 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 101100202924 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) tsp-2 gene Proteins 0.000 description 2
- 101100202932 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) tsp-4 gene Proteins 0.000 description 2
- 101100202938 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) tsp-5 gene Proteins 0.000 description 2
- ZRWPUFFVAOMMNM-UHFFFAOYSA-N Patulin Chemical compound OC1OCC=C2OC(=O)C=C12 ZRWPUFFVAOMMNM-UHFFFAOYSA-N 0.000 description 2
- XYFCBTPGUUZFHI-UHFFFAOYSA-N Phosphine Chemical compound P XYFCBTPGUUZFHI-UHFFFAOYSA-N 0.000 description 2
- 206010035226 Plasma cell myeloma Diseases 0.000 description 2
- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical compound C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 description 2
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 2
- ATUOYWHBWRKTHZ-UHFFFAOYSA-N Propane Chemical compound CCC ATUOYWHBWRKTHZ-UHFFFAOYSA-N 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 2
- 108090000951 RNA polymerase sigma 70 Proteins 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 101100208249 Rattus norvegicus Thbs4 gene Proteins 0.000 description 2
- 108010039491 Ricin Proteins 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 2
- 241000607142 Salmonella Species 0.000 description 2
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 2
- 241000607768 Shigella Species 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 108010092262 T-Cell Antigen Receptors Proteins 0.000 description 2
- 102100023935 Transmembrane glycoprotein NMB Human genes 0.000 description 2
- 102100021728 Transmembrane reductase CYB561D2 Human genes 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 230000007059 acute toxicity Effects 0.000 description 2
- 231100000403 acute toxicity Toxicity 0.000 description 2
- 239000000443 aerosol Substances 0.000 description 2
- 125000001931 aliphatic group Chemical group 0.000 description 2
- 150000005215 alkyl ethers Chemical class 0.000 description 2
- 150000001412 amines Chemical group 0.000 description 2
- 229960000723 ampicillin Drugs 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 230000000890 antigenic effect Effects 0.000 description 2
- 239000013011 aqueous formulation Substances 0.000 description 2
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 2
- 235000003704 aspartic acid Nutrition 0.000 description 2
- 125000004429 atom Chemical group 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 229960000686 benzalkonium chloride Drugs 0.000 description 2
- CADWTSSKOVRVJC-UHFFFAOYSA-N benzyl(dimethyl)azanium;chloride Chemical compound [Cl-].C[NH+](C)CC1=CC=CC=C1 CADWTSSKOVRVJC-UHFFFAOYSA-N 0.000 description 2
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 2
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 230000022534 cell killing Effects 0.000 description 2
- 238000012054 celltiter-glo Methods 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 125000003636 chemical group Chemical group 0.000 description 2
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 2
- 229960002173 citrulline Drugs 0.000 description 2
- 238000002648 combination therapy Methods 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 239000000356 contaminant Substances 0.000 description 2
- 229920001577 copolymer Polymers 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- KHAVLLBUVKBTBG-UHFFFAOYSA-N dec-9-enoic acid Chemical compound OC(=O)CCCCCCCC=C KHAVLLBUVKBTBG-UHFFFAOYSA-N 0.000 description 2
- 239000003405 delayed action preparation Substances 0.000 description 2
- 230000003111 delayed effect Effects 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 238000011033 desalting Methods 0.000 description 2
- 230000000368 destabilizing effect Effects 0.000 description 2
- 125000002228 disulfide group Chemical group 0.000 description 2
- 150000002019 disulfides Chemical class 0.000 description 2
- AMRJKAQTDDKMCE-UHFFFAOYSA-N dolastatin Chemical compound CC(C)C(N(C)C)C(=O)NC(C(C)C)C(=O)N(C)C(C(C)C)C(OC)CC(=O)N1CCCC1C(OC)C(C)C(=O)NC(C=1SC=CN=1)CC1=CC=CC=C1 AMRJKAQTDDKMCE-UHFFFAOYSA-N 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 239000002158 endotoxin Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- HQPMKSGTIOYHJT-UHFFFAOYSA-N ethane-1,2-diol;propane-1,2-diol Chemical compound OCCO.CC(O)CO HQPMKSGTIOYHJT-UHFFFAOYSA-N 0.000 description 2
- 230000007717 exclusion Effects 0.000 description 2
- 238000013265 extended release Methods 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 108020001507 fusion proteins Proteins 0.000 description 2
- 102000037865 fusion proteins Human genes 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- BZEUYEFVVDTLOD-UHFFFAOYSA-N hex-2-enamide Chemical compound CCCC=CC(N)=O BZEUYEFVVDTLOD-UHFFFAOYSA-N 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 238000004896 high resolution mass spectrometry Methods 0.000 description 2
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 2
- 125000000487 histidyl group Chemical class [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 230000005847 immunogenicity Effects 0.000 description 2
- 230000016784 immunoglobulin production Effects 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 238000001990 intravenous administration Methods 0.000 description 2
- 229910052740 iodine Inorganic materials 0.000 description 2
- 238000010829 isocratic elution Methods 0.000 description 2
- 229960000310 isoleucine Drugs 0.000 description 2
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 2
- 235000014705 isoleucine Nutrition 0.000 description 2
- 230000002147 killing effect Effects 0.000 description 2
- 235000005772 leucine Nutrition 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 238000004020 luminiscence type Methods 0.000 description 2
- 235000018977 lysine Nutrition 0.000 description 2
- 210000003712 lysosome Anatomy 0.000 description 2
- 230000001868 lysosomic effect Effects 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 125000005439 maleimidyl group Chemical group C1(C=CC(N1*)=O)=O 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 239000000594 mannitol Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- IDBIFFKSXLYUOT-UHFFFAOYSA-N netropsin Chemical compound C1=C(C(=O)NCCC(N)=N)N(C)C=C1NC(=O)C1=CC(NC(=O)CN=C(N)N)=CN1C IDBIFFKSXLYUOT-UHFFFAOYSA-N 0.000 description 2
- 229920002113 octoxynol Polymers 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 239000007800 oxidant agent Substances 0.000 description 2
- 238000007911 parenteral administration Methods 0.000 description 2
- WVDDGKGOMKODPV-ZQBYOMGUSA-N phenyl(114C)methanol Chemical compound O[14CH2]C1=CC=CC=C1 WVDDGKGOMKODPV-ZQBYOMGUSA-N 0.000 description 2
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 2
- 229960000502 poloxamer Drugs 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 238000002600 positron emission tomography Methods 0.000 description 2
- 229920001592 potato starch Polymers 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 230000002035 prolonged effect Effects 0.000 description 2
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 description 2
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 230000002285 radioactive effect Effects 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 238000009097 single-agent therapy Methods 0.000 description 2
- 238000002603 single-photon emission computed tomography Methods 0.000 description 2
- 239000001488 sodium phosphate Substances 0.000 description 2
- 229910000162 sodium phosphate Inorganic materials 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000002459 sustained effect Effects 0.000 description 2
- 238000013268 sustained release Methods 0.000 description 2
- 239000012730 sustained-release form Substances 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 238000004448 titration Methods 0.000 description 2
- 239000012929 tonicity agent Substances 0.000 description 2
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 2
- 239000012137 tryptone Substances 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 2
- 239000013603 viral vector Substances 0.000 description 2
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- UGJAEDFOKNAMQD-DVQDXYAYSA-N (-)-Falcarinol Natural products CCCCCCC\C=C\CC#CC#C[C@@H](O)C=C UGJAEDFOKNAMQD-DVQDXYAYSA-N 0.000 description 1
- KQODQNJLJQHFQV-UHFFFAOYSA-N (-)-hemiasterlin Natural products C1=CC=C2C(C(C)(C)C(C(=O)NC(C(=O)N(C)C(C=C(C)C(O)=O)C(C)C)C(C)(C)C)NC)=CN(C)C2=C1 KQODQNJLJQHFQV-UHFFFAOYSA-N 0.000 description 1
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 1
- OVJBOPBBHWOWJI-FYNXUGHNSA-N (2S)-2-[[(2S)-1-[(2S)-2-[[(aS,1R,3aS,4S,10S,16S,19R,22S,25S,28S,34S,37S,40R,45R,48S,51S,57S,60S,63S,69S,72S,75S,78S,85R,88S,91R,94S)-40-[[(2S)-6-amino-2-[[(2S)-2-[[(2S)-4-amino-2-[[(2S,3S)-2-[[(2S,3S)-2-[[(2S,3R)-2-amino-3-hydroxybutanoyl]amino]-3-methylpentanoyl]amino]-3-methylpentanoyl]amino]-4-oxobutanoyl]amino]-3-methylbutanoyl]amino]hexanoyl]amino]-25,48,78,88,94-pentakis(4-aminobutyl)-a-(2-amino-2-oxoethyl)-22,63,72-tris(3-amino-3-oxopropyl)-69-benzyl-37-[(1R)-1-hydroxyethyl]-34,60-bis(hydroxymethyl)-51,57,75-trimethyl-16-(2-methylpropyl)-3a-(2-methylsulfanylethyl)-2a,3,5a,9,15,18,21,24,27,33,36,39,47,50,53,56,59,62,65,68,71,74,77,80,87,90,93,96,99-nonacosaoxo-7a,8a,42,43,82,83-hexathia-1a,2,4a,8,14,17,20,23,26,32,35,38,46,49,52,55,58,61,64,67,70,73,76,79,86,89,92,95,98-nonacosazahexacyclo[43.35.25.419,91.04,8.010,14.028,32]nonahectane-85-carbonyl]amino]-3-(4-hydroxyphenyl)propanoyl]pyrrolidine-2-carbonyl]amino]-3-(1H-imidazol-5-yl)propanoic acid Chemical compound CC[C@H](C)[C@H](NC(=O)[C@@H](N)[C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@H]1CSSC[C@@H]2NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](Cc3ccccc3)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CCCCN)NC(=O)[C@@H]3CSSC[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@H](CSSC[C@H](NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H]4CCCN4C(=O)[C@H](CO)NC(=O)[C@@H](NC1=O)[C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N1CCC[C@H]1C(=O)N3)NC(=O)[C@H](CCCCN)NC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCSC)NC2=O)C(=O)N[C@@H](Cc1ccc(O)cc1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O OVJBOPBBHWOWJI-FYNXUGHNSA-N 0.000 description 1
- GTSCNQPMGNAJSH-YHFCJVPQSA-N (2S)-2-[[(2S)-2-[[(4R,7S,10S,16S,19R)-19-[[(3S,6S,9S,12S,15S,21R,26R,29S,32S,35S,38S,41S)-21-[[2-[[(1R,2aR,4S,7S,9aS,10S,12aS,13S,15aS,16S,18aS,19S,22S,25S,28S,31S,34S,37S,40S,43S,46S,52R,57R,60S,63S,66S,69S,72S,75S,81S,84S,87S,90S,93S,96S,99S)-7,63,90-tris(4-aminobutyl)-2a-[[(2S,3S)-2-[[(2S)-2-amino-5-carbamimidamidopentanoyl]amino]-3-methylpentanoyl]amino]-12a-(2-amino-2-oxoethyl)-25-(3-amino-3-oxopropyl)-13-benzyl-16,37,96-tris[(2S)-butan-2-yl]-19,28,46,72-tetrakis(3-carbamimidamidopropyl)-15a,31,43-tris(2-carboxyethyl)-9a,22,60,66-tetrakis[(1R)-1-hydroxyethyl]-40,84-bis(hydroxymethyl)-4,34,99-tris[(4-hydroxyphenyl)methyl]-93-(1H-imidazol-5-ylmethyl)-69,87-dimethyl-81-(2-methylpropyl)-10-(2-methylsulfanylethyl)-1a,2,5,8,8a,11,11a,14,14a,17,17a,20,20a,23,26,29,32,35,38,41,44,47,50,59,62,65,68,71,74,80,83,86,89,92,95,98-hexatriacontaoxo-18a-propan-2-yl-4a,5a,54,55-tetrathia-a,3,6,7a,9,10a,12,13a,15,16a,18,19a,21,24,27,30,33,36,39,42,45,48,51,58,61,64,67,70,73,79,82,85,88,91,94,97-hexatriacontazatricyclo[55.49.14.075,79]icosahectane-52-carbonyl]amino]acetyl]amino]-35-(3-amino-3-oxopropyl)-29-(2-carboxyethyl)-12,32-bis[(1R)-1-hydroxyethyl]-38-[(4-hydroxyphenyl)methyl]-3-(1H-indol-3-ylmethyl)-9-methyl-6-(2-methylsulfanylethyl)-2,5,8,11,14,20,28,31,34,37,40-undecaoxo-23,24-dithia-1,4,7,10,13,19,27,30,33,36,39-undecazatricyclo[39.3.0.015,19]tetratetracontane-26-carbonyl]amino]-16-(4-aminobutyl)-7-(3-carbamimidamidopropyl)-10-(carboxymethyl)-6,9,12,15,18-pentaoxo-1,2-dithia-5,8,11,14,17-pentazacycloicosane-4-carbonyl]amino]-4-amino-4-oxobutanoyl]amino]-6-aminohexanoic acid Chemical compound CC[C@H](C)[C@H](NC(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@H]1CSSC[C@@H]2NC(=O)[C@@H](NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CSSC[C@H](NC(=O)CNC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](Cc3ccc(O)cc3)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](NC(=O)[C@H](Cc3ccccc3)NC(=O)[C@H](CCSC)NC(=O)[C@H](CCCCN)NC(=O)[C@H](Cc3ccc(O)cc3)NC2=O)[C@@H](C)CC)[C@@H](C)O)[C@@H](C)CC)C(=O)NCC(=O)N[C@H]2CSSC[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](Cc3ccc(O)cc3)NC(=O)[C@@H]3CCCN3C(=O)[C@H](Cc3c[nH]c4ccccc34)NC(=O)[C@H](CCSC)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@@H]3CCCN3C2=O)[C@@H](C)O)[C@@H](C)O)C(=O)N[C@H]2CSSC[C@H](NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@H](CCCCN)NC2=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H]2CCCN2C(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](C)NC(=O)[C@H](CCCCN)NC(=O)[C@H](Cc2cnc[nH]2)NC(=O)[C@@H](NC(=O)[C@H](Cc2ccc(O)cc2)NC1=O)[C@@H](C)CC)[C@@H](C)O)[C@@H](C)O)C(C)C)[C@@H](C)O GTSCNQPMGNAJSH-YHFCJVPQSA-N 0.000 description 1
- YLCSLYZPLGQZJS-VDQHJUMDSA-N (2r)-2-acetamido-3-sulfanylpropanoic acid;(2s)-2,6-diaminohexanoic acid Chemical compound CC(=O)N[C@@H](CS)C(O)=O.NCCCC[C@H](N)C(O)=O YLCSLYZPLGQZJS-VDQHJUMDSA-N 0.000 description 1
- LDDMACCNBZAMSG-BDVNFPICSA-N (2r,3r,4s,5r)-3,4,5,6-tetrahydroxy-2-(methylamino)hexanal Chemical compound CN[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO LDDMACCNBZAMSG-BDVNFPICSA-N 0.000 description 1
- LGNCNVVZCUVPOT-FUVGGWJZSA-N (2s)-2-[[(2r,3r)-3-[(2s)-1-[(3r,4s,5s)-4-[[(2s)-2-[[(2s)-2-(dimethylamino)-3-methylbutanoyl]amino]-3-methylbutanoyl]-methylamino]-3-methoxy-5-methylheptanoyl]pyrrolidin-2-yl]-3-methoxy-2-methylpropanoyl]amino]-3-phenylpropanoic acid Chemical compound CC(C)[C@H](N(C)C)C(=O)N[C@@H](C(C)C)C(=O)N(C)[C@@H]([C@@H](C)CC)[C@H](OC)CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 LGNCNVVZCUVPOT-FUVGGWJZSA-N 0.000 description 1
- WOWDZACBATWTAU-FEFUEGSOSA-N (2s)-2-[[(2s)-2-(dimethylamino)-3-methylbutanoyl]amino]-n-[(3r,4s,5s)-1-[(2s)-2-[(1r,2r)-3-[[(1s,2r)-1-hydroxy-1-phenylpropan-2-yl]amino]-1-methoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl]-n,3-dimethylbutanamide Chemical compound CC(C)[C@H](N(C)C)C(=O)N[C@@H](C(C)C)C(=O)N(C)[C@@H]([C@@H](C)CC)[C@H](OC)CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C)[C@@H](O)C1=CC=CC=C1 WOWDZACBATWTAU-FEFUEGSOSA-N 0.000 description 1
- XMQUEQJCYRFIQS-YFKPBYRVSA-N (2s)-2-amino-5-ethoxy-5-oxopentanoic acid Chemical compound CCOC(=O)CC[C@H](N)C(O)=O XMQUEQJCYRFIQS-YFKPBYRVSA-N 0.000 description 1
- UGJAEDFOKNAMQD-MQNTZWLQSA-N (3S,9Z)-1,9-Heptadecadiene-4,6-diyn-3-ol Chemical compound CCCCCCC\C=C/CC#CC#C[C@@H](O)C=C UGJAEDFOKNAMQD-MQNTZWLQSA-N 0.000 description 1
- INAUWOVKEZHHDM-PEDBPRJASA-N (7s,9s)-6,9,11-trihydroxy-9-(2-hydroxyacetyl)-7-[(2r,4s,5s,6s)-5-hydroxy-6-methyl-4-morpholin-4-yloxan-2-yl]oxy-4-methoxy-8,10-dihydro-7h-tetracene-5,12-dione;hydrochloride Chemical compound Cl.N1([C@H]2C[C@@H](O[C@@H](C)[C@H]2O)O[C@H]2C[C@@](O)(CC=3C(O)=C4C(=O)C=5C=CC=C(C=5C(=O)C4=C(O)C=32)OC)C(=O)CO)CCOCC1 INAUWOVKEZHHDM-PEDBPRJASA-N 0.000 description 1
- KJBOIRGICUWVPF-MPFGFTFXSA-N (E,4S)-N-[(4-aminophenyl)methylsulfonyl]-4-[[(2S)-3,3-dimethyl-2-[[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino]butanoyl]-methylamino]-2,5-dimethylhex-2-enamide Chemical compound NC1=CC=C(CS(=O)(=O)NC(C(=C[C@H](C(C)C)N(C([C@H](C(C)(C)C)NC([C@H](C(C)(C2=CC=CC=C2)C)NC)=O)=O)C)C)=O)C=C1 KJBOIRGICUWVPF-MPFGFTFXSA-N 0.000 description 1
- VQGYYBCVUAWDGW-IDZRBWSNSA-N (E,4S)-N-[[4-(aminomethyl)phenyl]methylsulfonyl]-4-[[(2S)-3,3-dimethyl-2-[[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino]butanoyl]-methylamino]-2,5-dimethylhex-2-enamide Chemical compound NCC1=CC=C(CS(=O)(=O)NC(C(=C[C@H](C(C)C)N(C([C@H](C(C)(C)C)NC([C@H](C(C)(C2=CC=CC=C2)C)NC)=O)=O)C)C)=O)C=C1 VQGYYBCVUAWDGW-IDZRBWSNSA-N 0.000 description 1
- CNTMOLDWXSVYKD-PSRNMDMQSA-N (e,4s)-4-[[(2s)-3,3-dimethyl-2-[[(2s)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino]butanoyl]-methylamino]-2,5-dimethylhex-2-enoic acid Chemical compound OC(=O)C(/C)=C/[C@H](C(C)C)N(C)C(=O)[C@H](C(C)(C)C)NC(=O)[C@@H](NC)C(C)(C)C1=CC=CC=C1 CNTMOLDWXSVYKD-PSRNMDMQSA-N 0.000 description 1
- WJHZEAYXVJBOKT-VCXUBWRJSA-N (e,4s)-n-(4-aminophenyl)sulfonyl-4-[[(2s)-3,3-dimethyl-2-[[(2s)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino]butanoyl]-methylamino]-2,5-dimethylhex-2-enamide Chemical compound C(/[C@@H](N(C)C(=O)[C@@H](NC(=O)[C@@H](NC)C(C)(C)C=1C=CC=CC=1)C(C)(C)C)C(C)C)=C(/C)C(=O)NS(=O)(=O)C1=CC=C(N)C=C1 WJHZEAYXVJBOKT-VCXUBWRJSA-N 0.000 description 1
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- VYMHBQQZUYHXSS-UHFFFAOYSA-N 2-(3h-dithiol-3-yl)pyridine Chemical group C1=CSSC1C1=CC=CC=N1 VYMHBQQZUYHXSS-UHFFFAOYSA-N 0.000 description 1
- FBUTXZSKZCQABC-UHFFFAOYSA-N 2-amino-1-methyl-7h-purine-6-thione Chemical compound S=C1N(C)C(N)=NC2=C1NC=N2 FBUTXZSKZCQABC-UHFFFAOYSA-N 0.000 description 1
- MSWZFWKMSRAUBD-GASJEMHNSA-N 2-amino-2-deoxy-D-galactopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@H](O)[C@@H]1O MSWZFWKMSRAUBD-GASJEMHNSA-N 0.000 description 1
- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 description 1
- BFSVOASYOCHEOV-UHFFFAOYSA-N 2-diethylaminoethanol Chemical compound CCN(CC)CCO BFSVOASYOCHEOV-UHFFFAOYSA-N 0.000 description 1
- YIMDLWDNDGKDTJ-QLKYHASDSA-N 3'-deamino-3'-(3-cyanomorpholin-4-yl)doxorubicin Chemical compound N1([C@H]2C[C@@H](O[C@@H](C)[C@H]2O)O[C@H]2C[C@@](O)(CC=3C(O)=C4C(=O)C=5C=CC=C(C=5C(=O)C4=C(O)C=32)OC)C(=O)CO)CCOCC1C#N YIMDLWDNDGKDTJ-QLKYHASDSA-N 0.000 description 1
- JUMKLYDOIUWANQ-UHFFFAOYSA-N 3-[bis(2-bromoethyl)amino]-1-(4-phenylphenyl)propan-1-one Chemical compound C1=CC(C(=O)CCN(CCBr)CCBr)=CC=C1C1=CC=CC=C1 JUMKLYDOIUWANQ-UHFFFAOYSA-N 0.000 description 1
- IXHADCPJRQNDGG-UHFFFAOYSA-N 3-[bis(2-chloroethyl)amino]-1-(4-phenylphenyl)propan-1-one Chemical compound C1=CC(C(=O)CCN(CCCl)CCCl)=CC=C1C1=CC=CC=C1 IXHADCPJRQNDGG-UHFFFAOYSA-N 0.000 description 1
- QUHGSDZVAPFNLV-UHFFFAOYSA-N 4-[(5-acetamidofuran-2-carbonyl)amino]-n-[3-(dimethylamino)propyl]-1-propylpyrrole-2-carboxamide Chemical compound C1=C(C(=O)NCCCN(C)C)N(CCC)C=C1NC(=O)C1=CC=C(NC(C)=O)O1 QUHGSDZVAPFNLV-UHFFFAOYSA-N 0.000 description 1
- JHENVVKNLBODLB-UHFFFAOYSA-N 4-[[1-(2,5-dioxopyrrolidin-1-yl)-2h-pyridin-2-yl]disulfanyl]butanoic acid Chemical compound OC(=O)CCCSSC1C=CC=CN1N1C(=O)CCC1=O JHENVVKNLBODLB-UHFFFAOYSA-N 0.000 description 1
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 description 1
- LGZKGOGODCLQHG-CYBMUJFWSA-N 5-[(2r)-2-hydroxy-2-(3,4,5-trimethoxyphenyl)ethyl]-2-methoxyphenol Chemical compound C1=C(O)C(OC)=CC=C1C[C@@H](O)C1=CC(OC)=C(OC)C(OC)=C1 LGZKGOGODCLQHG-CYBMUJFWSA-N 0.000 description 1
- YXHLJMWYDTXDHS-IRFLANFNSA-N 7-aminoactinomycin D Chemical compound C[C@H]1OC(=O)[C@H](C(C)C)N(C)C(=O)CN(C)C(=O)[C@@H]2CCCN2C(=O)[C@@H](C(C)C)NC(=O)[C@H]1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=C(N)C=C3C(=O)N[C@@H]4C(=O)N[C@@H](C(N5CCC[C@H]5C(=O)N(C)CC(=O)N(C)[C@@H](C(C)C)C(=O)O[C@@H]4C)=O)C(C)C)=C3N=C21 YXHLJMWYDTXDHS-IRFLANFNSA-N 0.000 description 1
- 108700012813 7-aminoactinomycin D Proteins 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical class [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- NGNQZCDZXSOVQU-UHFFFAOYSA-N 8,16,18,26,34,36-hexahydroxyhentetracontane-2,6,10,14,24,28,32-heptone Chemical compound CCCCCC(O)CC(O)CC(=O)CCCC(=O)CC(O)CC(=O)CCCCCC(O)CC(O)CC(=O)CCCC(=O)CC(O)CC(=O)CCCC(C)=O NGNQZCDZXSOVQU-UHFFFAOYSA-N 0.000 description 1
- DBQULWBPIZECMN-UHFFFAOYSA-N 8,16,26,34,36-pentahydroxyhentetracontane-2,6,10,14,18,24,28,32-octone Chemical compound CCCCCC(O)CC(O)CC(=O)CCCC(=O)CC(O)CC(=O)CCCCCC(=O)CC(O)CC(=O)CCCC(=O)CC(O)CC(=O)CCCC(C)=O DBQULWBPIZECMN-UHFFFAOYSA-N 0.000 description 1
- 206010069754 Acquired gene mutation Diseases 0.000 description 1
- 208000031261 Acute myeloid leukaemia Diseases 0.000 description 1
- 241001436112 Aeropyrum coil-shaped virus Species 0.000 description 1
- 229930195730 Aflatoxin Natural products 0.000 description 1
- XWIYFDMXXLINPU-UHFFFAOYSA-N Aflatoxin G Chemical compound O=C1OCCC2=C1C(=O)OC1=C2C(OC)=CC2=C1C1C=COC1O2 XWIYFDMXXLINPU-UHFFFAOYSA-N 0.000 description 1
- 102100036826 Aldehyde oxidase Human genes 0.000 description 1
- 239000012116 Alexa Fluor 680 Substances 0.000 description 1
- 239000012118 Alexa Fluor 750 Substances 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 102100034044 All-trans-retinol dehydrogenase [NAD(+)] ADH1B Human genes 0.000 description 1
- 101710193111 All-trans-retinol dehydrogenase [NAD(+)] ADH4 Proteins 0.000 description 1
- 206010073478 Anaplastic large-cell lymphoma Diseases 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- 241000351920 Aspergillus nidulans Species 0.000 description 1
- 241000228245 Aspergillus niger Species 0.000 description 1
- 240000006439 Aspergillus oryzae Species 0.000 description 1
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 1
- 101000669426 Aspergillus restrictus Ribonuclease mitogillin Proteins 0.000 description 1
- 241000713826 Avian leukosis virus Species 0.000 description 1
- 108090001008 Avidin Proteins 0.000 description 1
- 108050001427 Avidin/streptavidin Proteins 0.000 description 1
- 102100038080 B-cell receptor CD22 Human genes 0.000 description 1
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 1
- 241000193738 Bacillus anthracis Species 0.000 description 1
- 101100404144 Bacillus subtilis (strain 168) nasD gene Proteins 0.000 description 1
- 101100249009 Bacillus subtilis (strain 168) rplK gene Proteins 0.000 description 1
- 241000151861 Barnettozyma salicaria Species 0.000 description 1
- ISNYUQWBWALXEY-UHFFFAOYSA-N Batrachotoxin Natural products C=1CC2(C3=CCC4C5(C)CCC(C4)(O)OC53C(O)C3)OCCN(C)CC32C=1C(C)OC(=O)C=1C(C)=CNC=1C ISNYUQWBWALXEY-UHFFFAOYSA-N 0.000 description 1
- 208000003508 Botulism Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 108700012439 CA9 Proteins 0.000 description 1
- 229960005532 CC-1065 Drugs 0.000 description 1
- 102100025221 CD70 antigen Human genes 0.000 description 1
- AUJXLBOHYWTPFV-BLWRDSOESA-N CS[C@H]1SC[C@H]2N(C)C(=O)[C@@H](C)NC(=O)[C@H](COC(=O)[C@@H](C(C)C)N(C)C(=O)[C@@H]1N(C)C(=O)[C@@H](C)NC(=O)[C@H](COC(=O)[C@@H](C(C)C)N(C)C2=O)NC(=O)c1cnc2ccccc2n1)NC(=O)c1cnc2ccccc2n1 Chemical compound CS[C@H]1SC[C@H]2N(C)C(=O)[C@@H](C)NC(=O)[C@H](COC(=O)[C@@H](C(C)C)N(C)C(=O)[C@@H]1N(C)C(=O)[C@@H](C)NC(=O)[C@H](COC(=O)[C@@H](C(C)C)N(C)C2=O)NC(=O)c1cnc2ccccc2n1)NC(=O)c1cnc2ccccc2n1 AUJXLBOHYWTPFV-BLWRDSOESA-N 0.000 description 1
- 101100327917 Caenorhabditis elegans chup-1 gene Proteins 0.000 description 1
- 241000282836 Camelus dromedarius Species 0.000 description 1
- 241000222122 Candida albicans Species 0.000 description 1
- 101710158575 Cap-specific mRNA (nucleoside-2'-O-)-methyltransferase Proteins 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 102100036369 Carbonic anhydrase 6 Human genes 0.000 description 1
- 102100024423 Carbonic anhydrase 9 Human genes 0.000 description 1
- 102100025475 Carcinoembryonic antigen-related cell adhesion molecule 5 Human genes 0.000 description 1
- 102100027473 Cartilage oligomeric matrix protein Human genes 0.000 description 1
- 102000005600 Cathepsins Human genes 0.000 description 1
- 108010084457 Cathepsins Proteins 0.000 description 1
- 101000997261 Centruroides margaritatus Potassium channel toxin alpha-KTx 2.2 Proteins 0.000 description 1
- 101000997271 Centruroides noxius Potassium channel toxin alpha-KTx 1.11 Proteins 0.000 description 1
- 108010023798 Charybdotoxin Proteins 0.000 description 1
- 241000195597 Chlamydomonas reinhardtii Species 0.000 description 1
- 108010049048 Cholera Toxin Proteins 0.000 description 1
- 102000009016 Cholera Toxin Human genes 0.000 description 1
- 102100039496 Choline transporter-like protein 4 Human genes 0.000 description 1
- 241001674013 Chrysosporium lucknowense Species 0.000 description 1
- FQVNSJQTSOVRKZ-JNRDBWBESA-N Cicutoxin Chemical compound CCC[C@@H](O)\C=C\C=C\C=C\C#CC#CCCCO FQVNSJQTSOVRKZ-JNRDBWBESA-N 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- 102100031162 Collagen alpha-1(XVIII) chain Human genes 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 1
- 241000699802 Cricetulus griseus Species 0.000 description 1
- 108700032819 Croton tiglium crotin II Proteins 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 101100208241 Danio rerio thbs3a gene Proteins 0.000 description 1
- 101100208245 Danio rerio thbs4b gene Proteins 0.000 description 1
- 101000723297 Dendroaspis polylepis polylepis Calciseptin Proteins 0.000 description 1
- 241000702421 Dependoparvovirus Species 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 239000004338 Dichlorodifluoromethane Substances 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- 235000017274 Diospyros sandwicensis Nutrition 0.000 description 1
- OFDNQWIFNXBECV-UHFFFAOYSA-N Dolastatin 10 Natural products CC(C)C(N(C)C)C(=O)NC(C(C)C)C(=O)N(C)C(C(C)CC)C(OC)CC(=O)N1CCCC1C(OC)C(C)C(=O)NC(C=1SC=CN=1)CC1=CC=CC=C1 OFDNQWIFNXBECV-UHFFFAOYSA-N 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 108010009858 Echinomycin Proteins 0.000 description 1
- 101001003194 Eleusine coracana Alpha-amylase/trypsin inhibitor Proteins 0.000 description 1
- 108010079505 Endostatins Proteins 0.000 description 1
- 241000991587 Enterovirus C Species 0.000 description 1
- 101000686777 Escherichia phage T7 T7 RNA polymerase Proteins 0.000 description 1
- 101710082714 Exotoxin A Proteins 0.000 description 1
- UGJAEDFOKNAMQD-UHFFFAOYSA-N Falcarinol Natural products CCCCCCCC=CCC#CC#CC(O)C=C UGJAEDFOKNAMQD-UHFFFAOYSA-N 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- UXDPXZQHTDAXOZ-UHFFFAOYSA-N Fumonisin B2 Natural products OC(=O)CC(C(O)=O)CC(=O)OC(C(C)CCCC)C(OC(=O)CC(CC(O)=O)C(O)=O)CC(C)CCCCCCC(O)CC(O)C(C)N UXDPXZQHTDAXOZ-UHFFFAOYSA-N 0.000 description 1
- 241000223218 Fusarium Species 0.000 description 1
- 241001149959 Fusarium sp. Species 0.000 description 1
- 241000567178 Fusarium venenatum Species 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 101150094690 GAL1 gene Proteins 0.000 description 1
- 102100028501 Galanin peptides Human genes 0.000 description 1
- 102100024637 Galectin-10 Human genes 0.000 description 1
- 102100039555 Galectin-7 Human genes 0.000 description 1
- 101001011019 Gallus gallus Gallinacin-10 Proteins 0.000 description 1
- 101001011021 Gallus gallus Gallinacin-12 Proteins 0.000 description 1
- 102100040004 Gamma-glutamylcyclotransferase Human genes 0.000 description 1
- JRZJKWGQFNTSRN-UHFFFAOYSA-N Geldanamycin Natural products C1C(C)CC(OC)C(O)C(C)C=C(C)C(OC(N)=O)C(OC)CCC=C(C)C(=O)NC2=CC(=O)C(OC)=C1C2=O JRZJKWGQFNTSRN-UHFFFAOYSA-N 0.000 description 1
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 1
- 101100100937 Giardia intestinalis TSP11 gene Proteins 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- HSRJKNPTNIJEKV-UHFFFAOYSA-N Guaifenesin Chemical compound COC1=CC=CC=C1OCC(O)CO HSRJKNPTNIJEKV-UHFFFAOYSA-N 0.000 description 1
- 101150009006 HIS3 gene Proteins 0.000 description 1
- 102100030595 HLA class II histocompatibility antigen gamma chain Human genes 0.000 description 1
- 108010072471 HTI-286 Proteins 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 208000017604 Hodgkin disease Diseases 0.000 description 1
- 208000021519 Hodgkin lymphoma Diseases 0.000 description 1
- 208000010747 Hodgkins lymphoma Diseases 0.000 description 1
- 101000928314 Homo sapiens Aldehyde oxidase Proteins 0.000 description 1
- 101000884305 Homo sapiens B-cell receptor CD22 Proteins 0.000 description 1
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 1
- 101000934356 Homo sapiens CD70 antigen Proteins 0.000 description 1
- 101000714525 Homo sapiens Carbonic anhydrase 6 Proteins 0.000 description 1
- 101000914324 Homo sapiens Carcinoembryonic antigen-related cell adhesion molecule 5 Proteins 0.000 description 1
- 101000725508 Homo sapiens Cartilage oligomeric matrix protein Proteins 0.000 description 1
- 101000851181 Homo sapiens Epidermal growth factor receptor Proteins 0.000 description 1
- 101100121078 Homo sapiens GAL gene Proteins 0.000 description 1
- 101000608772 Homo sapiens Galectin-7 Proteins 0.000 description 1
- 101000886680 Homo sapiens Gamma-glutamylcyclotransferase Proteins 0.000 description 1
- 101001082627 Homo sapiens HLA class II histocompatibility antigen gamma chain Proteins 0.000 description 1
- 101000934338 Homo sapiens Myeloid cell surface antigen CD33 Proteins 0.000 description 1
- 101000581981 Homo sapiens Neural cell adhesion molecule 1 Proteins 0.000 description 1
- 101000874179 Homo sapiens Syndecan-1 Proteins 0.000 description 1
- 101000634853 Homo sapiens T cell receptor alpha chain constant Proteins 0.000 description 1
- 101000633605 Homo sapiens Thrombospondin-2 Proteins 0.000 description 1
- 101000904724 Homo sapiens Transmembrane glycoprotein NMB Proteins 0.000 description 1
- 101000851376 Homo sapiens Tumor necrosis factor receptor superfamily member 8 Proteins 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- 101100273566 Humulus lupulus CCL10 gene Proteins 0.000 description 1
- 206010061598 Immunodeficiency Diseases 0.000 description 1
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 1
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 1
- 102000012745 Immunoglobulin Subunits Human genes 0.000 description 1
- 108010079585 Immunoglobulin Subunits Proteins 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- 241001138401 Kluyveromyces lactis Species 0.000 description 1
- 241000170280 Kluyveromyces sp. Species 0.000 description 1
- 241000235058 Komagataella pastoris Species 0.000 description 1
- LKDRXBCSQODPBY-AMVSKUEXSA-N L-(-)-Sorbose Chemical compound OCC1(O)OC[C@H](O)[C@@H](O)[C@@H]1O LKDRXBCSQODPBY-AMVSKUEXSA-N 0.000 description 1
- AHLPHDHHMVZTML-BYPYZUCNSA-N L-Ornithine Chemical compound NCCC[C@H](N)C(O)=O AHLPHDHHMVZTML-BYPYZUCNSA-N 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- 241000186610 Lactobacillus sp. Species 0.000 description 1
- 241000282838 Lama Species 0.000 description 1
- 208000032004 Large-Cell Anaplastic Lymphoma Diseases 0.000 description 1
- 108090001090 Lectins Proteins 0.000 description 1
- 102000004856 Lectins Human genes 0.000 description 1
- 101001049894 Leiurus hebraeus Potassium channel toxin alpha-KTx 5.1 Proteins 0.000 description 1
- 240000007472 Leucaena leucocephala Species 0.000 description 1
- 235000010643 Leucaena leucocephala Nutrition 0.000 description 1
- WEWBWVMTOYUPHH-QHAQEBJBSA-N Lotaustralin Chemical compound CC[C@](C)(C#N)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O WEWBWVMTOYUPHH-QHAQEBJBSA-N 0.000 description 1
- WEWBWVMTOYUPHH-HQPKYVJOSA-N Lotaustralin Natural products O([C@@](C#N)(CC)C)[C@H]1[C@@H](O)[C@@H](O)[C@H](O)[C@H](CO)O1 WEWBWVMTOYUPHH-HQPKYVJOSA-N 0.000 description 1
- 101150068888 MET3 gene Proteins 0.000 description 1
- 239000012515 MabSelect SuRe Substances 0.000 description 1
- 108700018351 Major Histocompatibility Complex Proteins 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 108090000015 Mesothelin Proteins 0.000 description 1
- 102000003735 Mesothelin Human genes 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 101100200099 Methanopyrus kandleri (strain AV19 / DSM 6324 / JCM 9639 / NBRC 100938) rps13 gene Proteins 0.000 description 1
- 244000302512 Momordica charantia Species 0.000 description 1
- 235000009811 Momordica charantia Nutrition 0.000 description 1
- 208000034578 Multiple myelomas Diseases 0.000 description 1
- 241000714177 Murine leukemia virus Species 0.000 description 1
- 208000033776 Myeloid Acute Leukemia Diseases 0.000 description 1
- 102100025243 Myeloid cell surface antigen CD33 Human genes 0.000 description 1
- 241000713883 Myeloproliferative sarcoma virus Species 0.000 description 1
- ZDZOTLJHXYCWBA-VCVYQWHSSA-N N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 ZDZOTLJHXYCWBA-VCVYQWHSSA-N 0.000 description 1
- GHAZCVNUKKZTLG-UHFFFAOYSA-N N-ethyl-succinimide Natural products CCN1C(=O)CCC1=O GHAZCVNUKKZTLG-UHFFFAOYSA-N 0.000 description 1
- HDFGOPSGAURCEO-UHFFFAOYSA-N N-ethylmaleimide Chemical compound CCN1C(=O)C=CC1=O HDFGOPSGAURCEO-UHFFFAOYSA-N 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 102100035486 Nectin-4 Human genes 0.000 description 1
- 101710043865 Nectin-4 Proteins 0.000 description 1
- 229930193140 Neomycin Natural products 0.000 description 1
- 108010042309 Netropsin Proteins 0.000 description 1
- 102100027347 Neural cell adhesion molecule 1 Human genes 0.000 description 1
- 241000221961 Neurospora crassa Species 0.000 description 1
- 101100022915 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) cys-11 gene Proteins 0.000 description 1
- 208000015914 Non-Hodgkin lymphomas Diseases 0.000 description 1
- QMGVPVSNSZLJIA-UHFFFAOYSA-N Nux Vomica Natural products C1C2C3C4N(C=5C6=CC=CC=5)C(=O)CC3OCC=C2CN2C1C46CC2 QMGVPVSNSZLJIA-UHFFFAOYSA-N 0.000 description 1
- 241000320412 Ogataea angusta Species 0.000 description 1
- 241001452677 Ogataea methanolica Species 0.000 description 1
- 241000489470 Ogataea trehalophila Species 0.000 description 1
- 241000826199 Ogataea wickerhamii Species 0.000 description 1
- AHLPHDHHMVZTML-UHFFFAOYSA-N Orn-delta-NH2 Natural products NCCCC(N)C(O)=O AHLPHDHHMVZTML-UHFFFAOYSA-N 0.000 description 1
- UTJLXEIPEHZYQJ-UHFFFAOYSA-N Ornithine Natural products OC(=O)C(C)CCCN UTJLXEIPEHZYQJ-UHFFFAOYSA-N 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 108010081690 Pertussis Toxin Proteins 0.000 description 1
- 241000530350 Phaffomyces opuntiae Species 0.000 description 1
- 241000529953 Phaffomyces thermotolerans Species 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- 101100413173 Phytolacca americana PAP2 gene Proteins 0.000 description 1
- 241000235062 Pichia membranifaciens Species 0.000 description 1
- 241000235061 Pichia sp. Species 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 101000762949 Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1) Exotoxin A Proteins 0.000 description 1
- 241001453299 Pseudomonas mevalonii Species 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- MUPFEKGTMRGPLJ-RMMQSMQOSA-N Raffinose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 MUPFEKGTMRGPLJ-RMMQSMQOSA-N 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 208000006265 Renal cell carcinoma Diseases 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- OWPCHSCAPHNHAV-UHFFFAOYSA-N Rhizoxin Natural products C1C(O)C2(C)OC2C=CC(C)C(OC(=O)C2)CC2CC2OC2C(=O)OC1C(C)C(OC)C(C)=CC=CC(C)=CC1=COC(C)=N1 OWPCHSCAPHNHAV-UHFFFAOYSA-N 0.000 description 1
- 241000191023 Rhodobacter capsulatus Species 0.000 description 1
- 241000191043 Rhodobacter sphaeroides Species 0.000 description 1
- 241000187562 Rhodococcus sp. Species 0.000 description 1
- 101100394989 Rhodopseudomonas palustris (strain ATCC BAA-98 / CGA009) hisI gene Proteins 0.000 description 1
- 241000190984 Rhodospirillum rubrum Species 0.000 description 1
- 241000714474 Rous sarcoma virus Species 0.000 description 1
- 108091007561 SLC44A4 Proteins 0.000 description 1
- 241000235070 Saccharomyces Species 0.000 description 1
- 101100434411 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) ADH1 gene Proteins 0.000 description 1
- 101100386089 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) MET17 gene Proteins 0.000 description 1
- 241000235088 Saccharomyces sp. Species 0.000 description 1
- 241000607149 Salmonella sp. Species 0.000 description 1
- 101100406813 Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720) pagC gene Proteins 0.000 description 1
- 206010039491 Sarcoma Diseases 0.000 description 1
- 101100022918 Schizosaccharomyces pombe (strain 972 / ATCC 24843) sua1 gene Proteins 0.000 description 1
- 101001049892 Scorpio palmatus Potassium channel toxin alpha-KTx 6.2 Proteins 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 101710142113 Serine protease inhibitor A3K Proteins 0.000 description 1
- 241000607720 Serratia Species 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- 108010079723 Shiga Toxin Proteins 0.000 description 1
- 241000607762 Shigella flexneri Species 0.000 description 1
- 241000607760 Shigella sonnei Species 0.000 description 1
- 241000607758 Shigella sp. Species 0.000 description 1
- 241000700584 Simplexvirus Species 0.000 description 1
- 241000713896 Spleen necrosis virus Species 0.000 description 1
- 102100035721 Syndecan-1 Human genes 0.000 description 1
- 102100029452 T cell receptor alpha chain constant Human genes 0.000 description 1
- 102100032272 T cell receptor delta constant Human genes 0.000 description 1
- 101710205572 T cell receptor delta constant Proteins 0.000 description 1
- 101150053966 THBS4 gene Proteins 0.000 description 1
- 229940123237 Taxane Drugs 0.000 description 1
- 108010055044 Tetanus Toxin Proteins 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- 101150116166 Thbs3 gene Proteins 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- 102100029529 Thrombospondin-2 Human genes 0.000 description 1
- 102100029524 Thrombospondin-3 Human genes 0.000 description 1
- 102100029219 Thrombospondin-4 Human genes 0.000 description 1
- UGPMCIBIHRSCBV-XNBOLLIBSA-N Thymosin beta 4 Chemical compound N([C@@H](CC(O)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O)C(=O)[C@@H]1CCCN1C(=O)[C@H](CCCCN)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(C)=O UGPMCIBIHRSCBV-XNBOLLIBSA-N 0.000 description 1
- 102100035000 Thymosin beta-4 Human genes 0.000 description 1
- 101710126507 Toxin 5 Proteins 0.000 description 1
- 101710170091 Transmembrane glycoprotein NMB Proteins 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 241000499912 Trichoderma reesei Species 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 102100036857 Tumor necrosis factor receptor superfamily member 8 Human genes 0.000 description 1
- MUPFEKGTMRGPLJ-UHFFFAOYSA-N UNPD196149 Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC2C(C(O)C(O)C(CO)O2)O)O1 MUPFEKGTMRGPLJ-UHFFFAOYSA-N 0.000 description 1
- 241000700618 Vaccinia virus Species 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 240000001866 Vernicia fordii Species 0.000 description 1
- 229940122803 Vinca alkaloid Drugs 0.000 description 1
- 241000370136 Wickerhamomyces pijperi Species 0.000 description 1
- 239000008351 acetate buffer Substances 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 125000003647 acryloyl group Chemical group O=C([*])C([H])=C([H])[H] 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 101150102866 adc1 gene Proteins 0.000 description 1
- 239000005409 aflatoxin Substances 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 229940100198 alkylating agent Drugs 0.000 description 1
- 239000002168 alkylating agent Substances 0.000 description 1
- 230000029936 alkylation Effects 0.000 description 1
- 238000005804 alkylation reaction Methods 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 108010001818 alpha-sarcin Proteins 0.000 description 1
- 230000006229 amino acid addition Effects 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 239000001166 ammonium sulphate Substances 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000000340 anti-metabolite Effects 0.000 description 1
- 230000002725 anti-mycoplasma Effects 0.000 description 1
- 230000001028 anti-proliverative effect Effects 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 229940100197 antimetabolite Drugs 0.000 description 1
- 239000002256 antimetabolite Substances 0.000 description 1
- 229940034982 antineoplastic agent Drugs 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000003125 aqueous solvent Substances 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 108010044540 auristatin Proteins 0.000 description 1
- 150000001541 aziridines Chemical class 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 238000003236 bicinchoninic acid assay Methods 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- RRKTZKIUPZVBMF-IBTVXLQLSA-N brucine Chemical compound O([C@@H]1[C@H]([C@H]2C3)[C@@H]4N(C(C1)=O)C=1C=C(C(=CC=11)OC)OC)CC=C2CN2[C@@H]3[C@]41CC2 RRKTZKIUPZVBMF-IBTVXLQLSA-N 0.000 description 1
- RRKTZKIUPZVBMF-UHFFFAOYSA-N brucine Natural products C1=2C=C(OC)C(OC)=CC=2N(C(C2)=O)C3C(C4C5)C2OCC=C4CN2C5C31CC2 RRKTZKIUPZVBMF-UHFFFAOYSA-N 0.000 description 1
- 239000008366 buffered solution Substances 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- DQXBYHZEEUGOBF-UHFFFAOYSA-N but-3-enoic acid;ethene Chemical compound C=C.OC(=O)CC=C DQXBYHZEEUGOBF-UHFFFAOYSA-N 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- HXCHCVDVKSCDHU-LULTVBGHSA-N calicheamicin Chemical compound C1[C@H](OC)[C@@H](NCC)CO[C@H]1O[C@H]1[C@H](O[C@@H]2C\3=C(NC(=O)OC)C(=O)C[C@](C/3=C/CSSSC)(O)C#C\C=C/C#C2)O[C@H](C)[C@@H](NO[C@@H]2O[C@H](C)[C@@H](SC(=O)C=3C(=C(OC)C(O[C@H]4[C@@H]([C@H](OC)[C@@H](O)[C@H](C)O4)O)=C(I)C=3C)OC)[C@@H](O)C2)[C@@H]1O HXCHCVDVKSCDHU-LULTVBGHSA-N 0.000 description 1
- 229930195731 calicheamicin Natural products 0.000 description 1
- 229940095731 candida albicans Drugs 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 230000009920 chelation Effects 0.000 description 1
- 238000012412 chemical coupling Methods 0.000 description 1
- 239000013611 chromosomal DNA Substances 0.000 description 1
- 229960004106 citric acid Drugs 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 239000013599 cloning vector Substances 0.000 description 1
- 201000010989 colorectal carcinoma Diseases 0.000 description 1
- LGZKGOGODCLQHG-UHFFFAOYSA-N combretastatin Natural products C1=C(O)C(OC)=CC=C1CC(O)C1=CC(OC)=C(OC)C(OC)=C1 LGZKGOGODCLQHG-UHFFFAOYSA-N 0.000 description 1
- 230000001447 compensatory effect Effects 0.000 description 1
- 238000013170 computed tomography imaging Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- CNVQLPPZGABUCM-LIGYZCPXSA-N ctx toxin Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](CO)C(=O)N[C@H]1CSSC[C@H]2C(=O)N[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@H]3CSSC[C@@H](C(N[C@@H](CC=4C5=CC=CC=C5NC=4)C(=O)N[C@@H](CO)C(=O)N[C@H](C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CCCNC(N)=N)NC3=O)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CO)C(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=3NC=NC=3)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N2)C(C)C)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H]([C@@H](C)O)NC1=O)=O)CCSC)C(C)C)[C@@H](C)O)NC(=O)[C@H]1NC(=O)CC1)C1=CC=CC=C1 CNVQLPPZGABUCM-LIGYZCPXSA-N 0.000 description 1
- 238000009109 curative therapy Methods 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
- 239000000824 cytostatic agent Substances 0.000 description 1
- JRLHSTVTOOELAF-KRWDZBQOSA-N dehydrofalcarinol Natural products O[C@@H](C=C)C#CC#CCC=CCCCCCC=C JRLHSTVTOOELAF-KRWDZBQOSA-N 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 229930191339 dianthin Natural products 0.000 description 1
- PXBRQCKWGAHEHS-UHFFFAOYSA-N dichlorodifluoromethane Chemical compound FC(F)(Cl)Cl PXBRQCKWGAHEHS-UHFFFAOYSA-N 0.000 description 1
- 235000019404 dichlorodifluoromethane Nutrition 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 1
- 230000003467 diminishing effect Effects 0.000 description 1
- 206010013023 diphtheria Diseases 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 230000006334 disulfide bridging Effects 0.000 description 1
- 229960003668 docetaxel Drugs 0.000 description 1
- 229930188854 dolastatin Natural products 0.000 description 1
- OFDNQWIFNXBECV-VFSYNPLYSA-N dolastatin 10 Chemical compound CC(C)[C@H](N(C)C)C(=O)N[C@@H](C(C)C)C(=O)N(C)[C@@H]([C@@H](C)CC)[C@H](OC)CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C=1SC=CN=1)CC1=CC=CC=C1 OFDNQWIFNXBECV-VFSYNPLYSA-N 0.000 description 1
- 108010045524 dolastatin 10 Proteins 0.000 description 1
- 229960000533 dornase alfa Drugs 0.000 description 1
- 108010067396 dornase alfa Proteins 0.000 description 1
- 229960004679 doxorubicin Drugs 0.000 description 1
- VQNATVDKACXKTF-XELLLNAOSA-N duocarmycin Chemical compound COC1=C(OC)C(OC)=C2NC(C(=O)N3C4=CC(=O)C5=C([C@@]64C[C@@H]6C3)C=C(N5)C(=O)OC)=CC2=C1 VQNATVDKACXKTF-XELLLNAOSA-N 0.000 description 1
- 229960005501 duocarmycin Drugs 0.000 description 1
- 229930184221 duocarmycin Natural products 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- XOPYFXBZMVTEJF-PDACKIITSA-N eleutherobin Chemical compound C(/[C@H]1[C@H](C(=CC[C@@H]1C(C)C)C)C[C@@H]([C@@]1(C)O[C@@]2(C=C1)OC)OC(=O)\C=C\C=1N=CN(C)C=1)=C2\CO[C@@H]1OC[C@@H](O)[C@@H](O)[C@@H]1OC(C)=O XOPYFXBZMVTEJF-PDACKIITSA-N 0.000 description 1
- XOPYFXBZMVTEJF-UHFFFAOYSA-N eleutherobin Natural products C1=CC2(OC)OC1(C)C(OC(=O)C=CC=1N=CN(C)C=1)CC(C(=CCC1C(C)C)C)C1C=C2COC1OCC(O)C(O)C1OC(C)=O XOPYFXBZMVTEJF-UHFFFAOYSA-N 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 210000001163 endosome Anatomy 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 108010028531 enomycin Proteins 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000011067 equilibration Methods 0.000 description 1
- 150000002170 ethers Chemical class 0.000 description 1
- 239000005038 ethylene vinyl acetate Substances 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 102000034287 fluorescent proteins Human genes 0.000 description 1
- 108091006047 fluorescent proteins Proteins 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000003008 fumonisin Substances 0.000 description 1
- UXDPXZQHTDAXOZ-STOIETHLSA-N fumonisin B2 Chemical compound OC(=O)C[C@@H](C(O)=O)CC(=O)O[C@H]([C@H](C)CCCC)[C@@H](OC(=O)C[C@@H](CC(O)=O)C(O)=O)C[C@@H](C)CCCCCC[C@@H](O)C[C@H](O)[C@H](C)N UXDPXZQHTDAXOZ-STOIETHLSA-N 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- QTQAWLPCGQOSGP-GBTDJJJQSA-N geldanamycin Chemical compound N1C(=O)\C(C)=C/C=C\[C@@H](OC)[C@H](OC(N)=O)\C(C)=C/[C@@H](C)[C@@H](O)[C@H](OC)C[C@@H](C)CC2=C(OC)C(=O)C=C1C2=O QTQAWLPCGQOSGP-GBTDJJJQSA-N 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 210000004602 germ cell Anatomy 0.000 description 1
- 229960002442 glucosamine Drugs 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 235000003969 glutathione Nutrition 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 125000005456 glyceride group Chemical group 0.000 description 1
- 125000003630 glycyl group Chemical group [H]N([H])C([H])([H])C(*)=O 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 229960002146 guaifenesin Drugs 0.000 description 1
- 229940093915 gynecological organic acid Drugs 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 108010057806 hemiasterlin Proteins 0.000 description 1
- 229930187626 hemiasterlin Natural products 0.000 description 1
- 239000008240 homogeneous mixture Substances 0.000 description 1
- 238000001794 hormone therapy Methods 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 210000004408 hybridoma Anatomy 0.000 description 1
- 150000007857 hydrazones Chemical class 0.000 description 1
- 239000000017 hydrogel Substances 0.000 description 1
- 229920001600 hydrophobic polymer Polymers 0.000 description 1
- 239000000819 hypertonic solution Substances 0.000 description 1
- 239000000815 hypotonic solution Substances 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 238000001361 intraarterial administration Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- UWKQSNNFCGGAFS-XIFFEERXSA-N irinotecan Chemical compound C1=C2C(CC)=C3CN(C(C4=C([C@@](C(=O)OC4)(O)CC)C=4)=O)C=4C3=NC2=CC=C1OC(=O)N(CC1)CCC1N1CCCCC1 UWKQSNNFCGGAFS-XIFFEERXSA-N 0.000 description 1
- FQVNSJQTSOVRKZ-UHFFFAOYSA-N isocicutoxin Natural products CCCC(O)C=CC=CC=CC#CC#CCCCO FQVNSJQTSOVRKZ-UHFFFAOYSA-N 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 235000007260 kalia Nutrition 0.000 description 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- BQINXKOTJQCISL-GRCPKETISA-N keto-neuraminic acid Chemical compound OC(=O)C(=O)C[C@H](O)[C@@H](N)[C@@H](O)[C@H](O)[C@H](O)CO BQINXKOTJQCISL-GRCPKETISA-N 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 239000002523 lectin Substances 0.000 description 1
- 108700009084 lexitropsin Proteins 0.000 description 1
- 108020001756 ligand binding domains Proteins 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 238000012792 lyophilization process Methods 0.000 description 1
- 239000012931 lyophilized formulation Substances 0.000 description 1
- 230000002132 lysosomal effect Effects 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- FSQQTNAZHBEJLS-UPHRSURJSA-N maleamic acid Chemical compound NC(=O)\C=C/C(O)=O FSQQTNAZHBEJLS-UPHRSURJSA-N 0.000 description 1
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 1
- 238000012531 mass spectrometric analysis of intact mass Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 108010082117 matrigel Proteins 0.000 description 1
- 238000002483 medication Methods 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 206010061289 metastatic neoplasm Diseases 0.000 description 1
- 150000002742 methionines Chemical class 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 108010010621 modeccin Proteins 0.000 description 1
- DASWEROEPLKSEI-UIJRFTGLSA-N monomethyl auristatin e Chemical compound CN[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(=O)N(C)[C@@H]([C@@H](C)CC)[C@H](OC)CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C)[C@@H](O)C1=CC=CC=C1 DASWEROEPLKSEI-UIJRFTGLSA-N 0.000 description 1
- MFRNYXJJRJQHNW-NARUGQRUSA-N monomethyl auristatin f Chemical compound CN[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(=O)N(C)C([C@@H](C)CC)[C@H](OC)CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 MFRNYXJJRJQHNW-NARUGQRUSA-N 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- DUWWHGPELOTTOE-UHFFFAOYSA-N n-(5-chloro-2,4-dimethoxyphenyl)-3-oxobutanamide Chemical compound COC1=CC(OC)=C(NC(=O)CC(C)=O)C=C1Cl DUWWHGPELOTTOE-UHFFFAOYSA-N 0.000 description 1
- 229960004927 neomycin Drugs 0.000 description 1
- 230000001613 neoplastic effect Effects 0.000 description 1
- CERZMXAJYMMUDR-UHFFFAOYSA-N neuraminic acid Natural products NC1C(O)CC(O)(C(O)=O)OC1C(O)C(O)CO CERZMXAJYMMUDR-UHFFFAOYSA-N 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 101150044129 nirB gene Proteins 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000002736 nonionic surfactant Substances 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000010534 nucleophilic substitution reaction Methods 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 238000012634 optical imaging Methods 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 229960003104 ornithine Drugs 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 238000002638 palliative care Methods 0.000 description 1
- 201000002528 pancreatic cancer Diseases 0.000 description 1
- 208000008443 pancreatic carcinoma Diseases 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 230000005298 paramagnetic effect Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 108010076042 phenomycin Proteins 0.000 description 1
- 229910000073 phosphorus hydride Inorganic materials 0.000 description 1
- 229940012957 plasmin Drugs 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920001200 poly(ethylene-vinyl acetate) Polymers 0.000 description 1
- 229920000747 poly(lactic acid) Polymers 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 229920005862 polyol Polymers 0.000 description 1
- 150000003077 polyols Chemical class 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000000244 polyoxyethylene sorbitan monooleate Substances 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920002503 polyoxyethylene-polyoxypropylene Polymers 0.000 description 1
- 229920005606 polypropylene copolymer Polymers 0.000 description 1
- 229940068977 polysorbate 20 Drugs 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 229940068968 polysorbate 80 Drugs 0.000 description 1
- 230000004481 post-translational protein modification Effects 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 150000003141 primary amines Chemical class 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 239000001294 propane Substances 0.000 description 1
- 239000003380 propellant Substances 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 238000000159 protein binding assay Methods 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 231100000654 protein toxin Toxicity 0.000 description 1
- 229950010131 puromycin Drugs 0.000 description 1
- WHMDPDGBKYUEMW-UHFFFAOYSA-N pyridine-2-thiol Chemical compound SC1=CC=CC=N1 WHMDPDGBKYUEMW-UHFFFAOYSA-N 0.000 description 1
- 239000002510 pyrogen Substances 0.000 description 1
- YUOCYTRGANSSRY-UHFFFAOYSA-N pyrrolo[2,3-i][1,2]benzodiazepine Chemical class C1=CN=NC2=C3C=CN=C3C=CC2=C1 YUOCYTRGANSSRY-UHFFFAOYSA-N 0.000 description 1
- AUJXLBOHYWTPFV-UHFFFAOYSA-N quinomycin A Natural products CN1C(=O)C(C)NC(=O)C(NC(=O)C=2N=C3C=CC=CC3=NC=2)COC(=O)C(C(C)C)N(C)C(=O)C2N(C)C(=O)C(C)NC(=O)C(NC(=O)C=3N=C4C=CC=CC4=NC=3)COC(=O)C(C(C)C)N(C)C(=O)C1CSC2SC AUJXLBOHYWTPFV-UHFFFAOYSA-N 0.000 description 1
- UOWVMDUEMSNCAV-WYENRQIDSA-N rachelmycin Chemical compound C1([C@]23C[C@@H]2CN1C(=O)C=1NC=2C(OC)=C(O)C4=C(C=2C=1)CCN4C(=O)C1=CC=2C=4CCN(C=4C(O)=C(C=2N1)OC)C(N)=O)=CC(=O)C1=C3C(C)=CN1 UOWVMDUEMSNCAV-WYENRQIDSA-N 0.000 description 1
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
- 230000008707 rearrangement Effects 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 238000010188 recombinant method Methods 0.000 description 1
- 238000010405 reoxidation reaction Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 230000001177 retroviral effect Effects 0.000 description 1
- OWPCHSCAPHNHAV-LMONGJCWSA-N rhizoxin Chemical compound C/C([C@H](OC)[C@@H](C)[C@@H]1C[C@H](O)[C@]2(C)O[C@@H]2/C=C/[C@@H](C)[C@]2([H])OC(=O)C[C@@](C2)(C[C@@H]2O[C@H]2C(=O)O1)[H])=C\C=C\C(\C)=C\C1=COC(C)=N1 OWPCHSCAPHNHAV-LMONGJCWSA-N 0.000 description 1
- 101150049069 rpsM gene Proteins 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- MXWDLLUGULWYIQ-BFRWRHKQSA-N scyllatoxin Chemical compound C([C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1N=CNC=1)C(N)=O)NC(=O)[C@H](C)N)C1=CC=CC=C1 MXWDLLUGULWYIQ-BFRWRHKQSA-N 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 125000003607 serino group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C(O[H])([H])[H] 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 229940115939 shigella sonnei Drugs 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 238000001542 size-exclusion chromatography Methods 0.000 description 1
- IYKSHBADGOZWIF-UTPLJIOFSA-N slotoxin Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](C(C)C)C(O)=O)NC(=O)[C@@H](N)[C@@H](C)O)C1=CC=CC=C1 IYKSHBADGOZWIF-UTPLJIOFSA-N 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 239000012475 sodium chloride buffer Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 238000010532 solid phase synthesis reaction Methods 0.000 description 1
- 230000037439 somatic mutation Effects 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000008174 sterile solution Substances 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 125000004434 sulfur atom Chemical group 0.000 description 1
- 230000020382 suppression by virus of host antigen processing and presentation of peptide antigen via MHC class I Effects 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- DKPFODGZWDEEBT-QFIAKTPHSA-N taxane Chemical class C([C@]1(C)CCC[C@@H](C)[C@H]1C1)C[C@H]2[C@H](C)CC[C@@H]1C2(C)C DKPFODGZWDEEBT-QFIAKTPHSA-N 0.000 description 1
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 1
- 229940118376 tetanus toxin Drugs 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- UWHCKJMYHZGTIT-UHFFFAOYSA-N tetraethylene glycol Chemical compound OCCOCCOCCOCCO UWHCKJMYHZGTIT-UHFFFAOYSA-N 0.000 description 1
- CFMYXEVWODSLAX-UHFFFAOYSA-N tetrodotoxin Natural products C12C(O)NC(=N)NC2(C2O)C(O)C3C(CO)(O)C1OC2(O)O3 CFMYXEVWODSLAX-UHFFFAOYSA-N 0.000 description 1
- 108010079996 thymosin beta(4) Proteins 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- UCFGDBYHRUNTLO-QHCPKHFHSA-N topotecan Chemical compound C1=C(O)C(CN(C)C)=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 UCFGDBYHRUNTLO-QHCPKHFHSA-N 0.000 description 1
- 229960000303 topotecan Drugs 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000005030 transcription termination Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- LZAJKCZTKKKZNT-PMNGPLLRSA-N trichothecene Chemical compound C12([C@@]3(CC[C@H]2OC2C=C(CCC23C)C)C)CO1 LZAJKCZTKKKZNT-PMNGPLLRSA-N 0.000 description 1
- 229930013292 trichothecene Natural products 0.000 description 1
- ISNYUQWBWALXEY-OMIQOYQYSA-N tsg6xhx09r Chemical compound O([C@@H](C)C=1[C@@]23CN(C)CCO[C@]3(C3=CC[C@H]4[C@]5(C)CC[C@@](C4)(O)O[C@@]53[C@H](O)C2)CC=1)C(=O)C=1C(C)=CNC=1C ISNYUQWBWALXEY-OMIQOYQYSA-N 0.000 description 1
- 230000005748 tumor development Effects 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 150000003668 tyrosines Chemical class 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 210000003501 vero cell Anatomy 0.000 description 1
- 239000003643 water by type Substances 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
- C07K16/32—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products of oncogenes
-
- 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
-
- 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
-
- 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
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
- A61K49/0039—Coumarin dyes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
- A61K49/0058—Antibodies
-
- 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
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
- C07K2317/14—Specific host cells or culture conditions, e.g. components, pH or temperature
-
- 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
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/52—Constant or Fc region; Isotype
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/52—Constant or Fc region; Isotype
- C07K2317/522—CH1 domain
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/52—Constant or Fc region; Isotype
- C07K2317/53—Hinge
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
-
- 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
- C07K2317/90—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
Definitions
- ADCs antibody drug conjugates
- ADCs are generally composed of an antibody chemically or enzymatically coupled to a drug (e.g., a cytotoxic drug), often via a linker.
- ADCs are typically designed to be stable in circulation and to effect intracellular drug release following antigen-specific binding and, in some instances, internalization of the ADC. Because ADCs may be designed to deliver a "payload" (such as a cytotoxic drug) to the cellular target, the efficiency of target cell modulation by the agent (e.g., target cell killing) may be much greater in the context of an ADC as compared to the
- ADCs that provide for conjugation of a drug payload at selected site(s) in the antibody are of interest, for a number of reasons, including the desire for homogeneity of product in an antibody drug conjugate preparation.
- some groups have explored amino acid substitution at specific sites within antibodies in an attempt to facilitate site-specific payload attachment while maintaining antibody structure and function. For example, Shen et al., have systematically examined cysteine substitution at various positions within antibody heavy and light chains to reveal the impact of site selection on conjugate stability (e.g., Nat. Biotech., 30: 184-189, 2012). Notably, these studies have revealed that solvent accessibility of the site of attachment on an antibody can negatively impact the stability of a resulting ADC.
- the antigen specificity of antibodies has also been exploited to provide diagnostic and imaging tools that incorporate labeled agents and recognize epitopes and/or cells that inform diagnostic or prognostic determinations and the course of therapy.
- diagnostic and prognostic tools rely on the affinity of tool antibodies for antigen detection and rely on the retention of labeled agents by tool antibodies for specific signaling. Accordingly, as with ADCs, locating or creating sites amenable to label attachment in a site-specific manner without compromising antibody affinity or the stability of resultant conjugates is highly desired.
- the present disclosure provides antibody light chain polypeptides that include a C- terminal amino acid extension, as well as antibodies and antibody conjugates containing such modified light chain polypeptides, where the C-terminal extension includes one or more cysteine residues.
- Conjugates that include an antibody of the present disclosure conjugated to an agent via the cysteine residue of the C-terminal amino acid extension are also provided.
- the present disclosure further provides nucleic acids encoding an antibody light chain polypeptide that includes a C-terminal amino acid extension including a cysteine residue.
- Pharmaceutical compositions including the antibodies or conjugates of the present disclosure are also provided, as are methods of making and use of the antibodies and conjugates of the present disclosure.
- the present disclosure provides an antibody including a light chain polypeptide that includes a C-terminal amino acid extension including a cysteine residue.
- the present disclosure provides an antibody that includes a light chain polypeptide including a C-terminal amino acid extension that includes a cysteine residue, where the C-terminal amino acid extension does not specifically bind antigen (e.g., the extension does not include an antigen-binding portion of an antibody or an antigen-binding portion of an antibody fragment).
- the present disclosure provides an antibody that includes at least one monoepitopic antigen-binding dimer, where the monoepitopic antigen-binding dimer includes a heavy chain polypeptide and a light chain polypeptide that includes a C-terminal amino acid extension, which extension includes a cysteine residue.
- Each of the two monoepitopic dimers may bind to the same epitope. In other aspects, each of the two monoepitopic dimers binds a different epitope.
- the C-terminal amino acid extension of any of the antibodies summarized above includes an amino acid spacer that does not include a cysteine residue.
- the spacer is from 1 to 30 amino acids, from 3 to 20 amino acids, or from 4 to 17 amino acids.
- the spacer includes a glycine (G) residue and a serine (S) residue.
- the spacer may consist of one or more glycine (G) residues and one or more serine (S) residues.
- Such a spacer optionally has the sequence GGGS.
- the extension includes endogenous human amino acid sequences or modified human amino acid sequences. These may include human antibody hinge region sequences, T-cell receptor sequences or other human sequences.
- the extension may include extracellular protein amino acid sequences and/or amino acid sequences of extracellular domains of proteins present on a cell surface.
- the extension includes an endogenous human amino acid sequence that includes one or more naturally occurring cysteine residues. These may include human antibody hinge region sequences, T-cell receptor sequences or other human protein cysteine containing sequences.
- the extension may include extracellular protein amino acid sequences and/or amino acid sequences of extracellular domains of proteins present on a cell surface.
- the extension includes a modified human amino acid sequence wherein one or more cysteines has been introduced into the endogenous human amino acid sequence by insertion or substitution.
- a cysteine is present between each of the spacers.
- at least two of the spacers may be contiguous, e.g., at least two of the spacers in the C-terminal amino acid extension do not include any amino acids (e.g., any cysteines) between the spacers.
- the C-terminal amino acid extension terminates in a cysteine.
- a cysteine within the C-terminal amino acid extension of an antibody of the present disclosure includes a reduced sulfhydryl group.
- the antibody includes an agent conjugated to the cysteine residue of the C-terminal amino acid extension.
- the agent is directly conjugated to the cysteine residue of the C- terminal amino acid extension.
- the agent is indirectly conjugated to the cysteine residue of the C-terminal amino acid extension via a linker.
- the agent is preferentially conjugated to the cysteine of the C-terminal amino acid extension of the light chain over a cysteine residue outside the C-terminal amino acid extension.
- the agent is exclusively conjugated to the antibody via the cysteine of the C- terminal amino acid extension of the light chain of the antibody.
- the agent is a therapeutic agent (e.g., a cytotoxic agent) or a labeling agent (e.g., an in vivo imaging agent).
- the C-terminal amino acid extension includes two or more cysteines each conjugated to an agent independently selected from a therapeutic agent and a labeling agent.
- two or more agents are independently directly or indirectly conjugated to two or more cysteine residues of the C-terminal amino acid extension.
- the agents are preferentially conjugated to the cysteines of the C-terminal amino acid extension of the light chain over a cysteine residue outside the C-terminal amino acid extension.
- the agents are exclusively conjugated to the antibody via the cysteines of the C-terminal amino acid extension of the light chain of the antibody.
- Antibodies of the present disclosure may be an antibody or binding fragment thereof.
- the antibody may be an IgG, Fab, F(ab')2, Fab', Fv, ScFv, bispecific antibody, or the like.
- conjugates include an antibody including a light chain polypeptide that includes a C-terminal amino acid extension including a cysteine residue.
- the conjugates include an antibody that includes a light chain polypeptide including a C-terminal amino acid extension that includes a cysteine residue, where the C-terminal amino acid extension does not specifically bind antigen (e.g., the extension does not include an antigen-binding portion of an antibody or an antigen-binding portion of an antibody fragment).
- the conjugates include an antibody that includes at least one monoepitopic antigen-binding dimer, where the monoepitopic antigen-binding dimer includes a heavy chain polypeptide and a light chain polypeptide that includes a C-terminal amino acid extension, which extension includes a cysteine residue.
- the antibodies of such conjugates may include two monoepitopic antigen-binding dimers. Each of the two monoepitopic dimers may bind to the same epitope. In other aspects, each of the two monoepitopic dimers binds a different epitope.
- the conjugates further include an agent conjugated to the antibody via the cysteine residue of the C-terminal amino acid extension.
- the antibody and agent of the conjugates of the present disclosure may have any of the antibody and agent features summarized above or described in the detailed description and examples section hereinbelow, and may be conjugated using the conjugation strategies described herein or any other suitable strategy that provides for the same conjugation results.
- nucleic acids that encode all or a portion of the antibodies of the present disclosure.
- a nucleic acid that encodes an antibody light chain polypeptide including a C-terminal amino acid extension including a cysteine residue.
- the C-terminal amino acid extension may include any of the features summarized above with respect to the antibodies of the present disclosure, and as described in the detailed description and examples section hereinbelow.
- Vectors that include such nucleic acids, and host cells e.g., prokaryotic or eukaryotic host cells
- host cells e.g., prokaryotic or eukaryotic host cells
- the present disclosure provides pharmaceutical compositions.
- the pharmaceutical compositions include any of the antibodies or conjugates summarized above with respect to the antibodies and conjugates of the present disclosure, and as described in the detailed description and examples section
- methods that include administering to a patient in need thereof a therapeutically effective amount of any of the pharmaceutical compositions, the antibodies or conjugates summarized above with respect to the antibodies and conjugates of the present disclosure, and as described in the detailed description and examples section hereinbelow.
- Methods of making a light chain of an antibody are also provided. Such methods include expressing in a host cell a nucleic acid encoding an antibody light chain polypeptide including a C-terminal amino acid extension that includes a cysteine residue. In certain aspects, the methods further include reducing the sulfhydryl group of the cysteine residue in the C-terminal amino acid extension.
- the methods comprise the preferential (or "biased") reduction of the sulfhydryl group of the cysteine residue in the C-terminal amino acid extension over the reduction of cysteine residues outside the C-terminal amino acid extension. In one embodiment, the methods comprise the exclusive reduction of the sulfhydryl group of the cysteine residue in the C-terminal amino acid extension over the reduction of cysteine residues outside the C-terminal amino acid extension.
- the C-terminal amino acid extension includes two or more cysteine residues.
- the methods further include reducing the sulfhydryl groups of the cysteine residues in the C-terminal amino acid extension.
- the methods comprise the preferential (or "biased") reduction of the sulfhydryl groups of the cysteine residues in the C-terminal amino acid extension over the reduction of cysteine residues outside the C- terminal amino acid extension.
- the methods comprise the exclusive reduction of the sulfhydryl groups of the cysteine residues in the C-terminal amino acid extension over the reduction of cysteine residues outside the C-terminal amino acid extension.
- aspects of the present disclosure include methods of making antibody conjugates.
- the methods include conjugating an agent to an antibody including a light chain polypeptide that includes a C-terminal amino acid extension including a cysteine residue, where the agent is conjugated to the cysteine residue of the C-terminal amino acid extension.
- the methods of making antibody conjugates may further include reducing the sulfhydryl group of the cysteine in the C-terminal amino acid extension prior to conjugating the agent to the antibody.
- the conjugating includes crosslinking the agent to the reduced sulfhydryl group using maleimide reaction chemistry, haloacetyl reaction chemistry, vinyl sulfone reaction chemistry or pyridyl disulfide reaction chemistry.
- the agent that is conjugated to the cysteine residue of the C-terminal amino acid extension is a therapeutic agent or a labeling agent.
- the agent is conjugated to the cysteine residue of the C-terminal amino acid extension preferentially over cysteine residues outside the C-terminal amino acid extension.
- the agent is conjugated to the cysteine residue of the C-terminal amino acid extension and not to any cysteine residues outside the C-terminal amino acid extension.
- the C-terminal amino acid extension includes two or more cysteine residues, and two or more agents are conjugated to the cysteine residues of the C-terminal amino acid extension.
- the methods of making such antibody conjugates may further include reducing the sulfhydryl groups of the cysteines in the C-terminal amino acid extension prior to conjugating the agents to the antibody.
- the conjugating includes crosslinking the agents to the reduced sulfhydryl groups using maleimide reaction chemistry, haloacetyl reaction chemistry, vinyl sulfone reaction chemistry or pyridyl disulfide reaction chemistry.
- the agents that are conjugated to the cysteine residues of the C- terminal amino acid extensions are therapeutic agents and/or labeling agents. In one
- the agents are conjugated to the cysteine residues of the C-terminal amino acid extension preferentially over cysteine residues outside the C-terminal amino acid extension. In one embodiment, the agents are conjugated to the cysteine residues of the C-terminal amino acid extension and not to any cysteine residues outside the C-terminal amino acid extension.
- FIG. 1 is a schematic illustration of an antibody that includes a C-terminal light chain extension according to one embodiment of the present disclosure.
- FIG. 2 is a schematic illustration of a conjugate according to one embodiment of the present disclosure.
- FIG. 3 provides cancer cell viability data for two example antibody conjugates according to embodiments of the present disclosure.
- FIG. 4 Panels A and B show antibody binding data for unconjugated antibodies according to certain embodiments of the present disclosure.
- FIG. 5 Panels A and B show antibody binding data for antibody-drug conjugates according to certain embodiments of the present disclosure.
- FIG. 8 shows in vivo tumor volume change over time in mice administered antibodies or antibody conjugates according to certain aspects of the present disclosure.
- Panels A-C provide size exclusion chromatography-mass spectrometry (SEC-
- Panels A-C provide SEC-MS data for an antibody (T-VLcys2) having a C- terminal light chain extension according to an embodiment of the present disclosure.
- Panels A and B provide SEC-MS data for an antibody (T-VLcys4) having a C- terminal light chain extension according to an embodiment of the present disclosure.
- Panels A and B provide SEC-MS data for an antibody (T-SP2) having a C- terminal light chain extension according to an embodiment of the present disclosure.
- Panels A-C provide SEC-MS data for an antibody (T-SP3) having a C-terminal light chain extension according to an embodiment of the present disclosure.
- Panels A and B provide SEC-MS data for an antibody (T-SP5) having a C- terminal light chain extension according to an embodiment of the present disclosure.
- Panels A-C provide SEC-MS data for an antibody (T-SP6) having a C-terminal light chain extension according to an embodiment of the present disclosure.
- Panels A-C provide SEC-MS data for an antibody (T-SP7) having a C-terminal light chain extension according to an embodiment of the present disclosure.
- Panels A-C provide SEC-MS data for an antibody (T-SP10) having a C-terminal light chain extension according to an embodiment of the present disclosure.
- Panels A-C provide SEC-MS data for an antibody (T-SP11) having a C-terminal light chain extension according to an embodiment of the present disclosure.
- FIG. 20 shows an HIC chromatograph of conjugation reaction products for Tsp2-Toxin 3.
- the average drug loading value was 1.92.
- FIG. 21 shows an HIC chromatograph of conjugation reaction products for Tsp3-Toxin 3.
- FIG. 22 shows an HIC chromatograph of conjugation reaction products for Tsp4-Toxin 3.
- the average drug loading value was 1.16.
- FIG. 23 shows an HIC chromatograph of conjugation reaction products for Tsp5-Toxin 3.
- the average drug loading value was 1.46.
- FIG. 24 shows an HIC chromatograph of conjugation reaction products for Tsp6-Toxin 3.
- the average drug loading value was 0.98.
- FIG. 25 shows an HIC chromatograph of conjugation reaction products for Tsp9-Toxin 3.
- the average drug loading value was 1.64.
- FIG. 26 shows an HIC chromatograph of conjugation reaction products for TsplO-Toxin 3 (larger scale). The average drug loading value was 2.0.
- FIG. 27 shows an HIC chromatograph of conjugation reaction products for Tspl 1 -Toxin 3.
- the average drug loading value was 2.66.
- FIG. 28 shows an HIC chromatograph of conjugation reaction products for TVLCysl- Toxin3.
- the average drug loading value was 2.66.
- FIG. 29 shows an HIC chromatograph of conjugation reaction products for TVLCys2-
- the average drug loading value was 0.22.
- FIG. 30 shows an HIC chromatograph of conjugation reaction products for TVLCys4- Toxin3.
- the average drug loading value was 0.70.
- FIG. 31 shows an HIC chromatograph of conjugation reaction products for TsplO-Toxin 3 (larger scale). The average drug loading value was 2.12.
- FIG. 32 shows an HIC chromatograph of conjugation reaction products for TsplO-Toxin 4 (larger scale).
- the average drug loading value was 3.76, where average attachments was 1.88.
- FIG. 33 shows an HIC chromatograph of conjugation reaction products for TsplO-Toxin 1 (larger scale). The average drug loading value was 1.94.
- FIG. 34 shows an HIC chromatograph of conjugation reaction products for Tsp4-Toxin 3
- FIG. 35 shows an HIC chromatograph of conjugation reaction products for Tsp6-Toxin 3 (larger scale). The average drug loading value was 1.82.
- FIG. 36 shows an HIC chromatograph of conjugation reaction products for T-Toxin 3.
- the average drug loading value was 0.16.
- FIG. 37 shows an HIC chromatograph of conjugation reaction products for BsplO- MCvcPABC-MMAE, where "B” is an abbreviation for Brentuximab anti-CD30 antibody.
- the average drug loading value was 2.12.
- FIG. 38 shows an HIC chromatograph of conjugation reaction products for BsplO-Toxin 4.
- the average drug loading value was 1.96.
- FIG. 39 shows an HIC chromatograph of conjugation reaction products for BsplO-Toxin
- the average drug loading value was 2.18.
- FIG. 40 shows an HIC chromatograph of conjugation reaction products for BsplO-Toxin 3.
- the average drug loading value was 1.98.
- FIG. 41 shows an HIC chromatograph of conjugation reaction products for BsplO-Toxin
- FIG. 42 shows a plot of in vitro cell proliferation assay results with Her2 expressing HCC1954 cells treated with Tsp4-Toxin3, Tsp3-Toxin3, Tsp2-Toxin3, and Free Toxinl .
- FIG. 43 shows a plot of in vitro cell proliferation assay results with Her2 expressing HCC1954 cells treated with Tsp5-Toxin3, Tsp6-Toxin3, Tsp9-Toxin3, and Free Toxinl .
- FIG. 44 shows a plot of in vitro cell proliferation assay results with Her2 expressing HCC1954 cells treated with Tspl0-Toxin3, Tspl l-Toxin3, TVLCysl-Toxin3, and Free Toxinl .
- FIG. 45 shows a plot of in vitro cell proliferation assay results with HER2 antigen negative Jurkat cells treated with Tsp4-Toxin3, Tsp3-Toxin3, Tsp2-Toxin3, and Free Toxinl .
- FIG. 46 shows a plot of in vitro cell proliferation assay results with HER2 antigen negative Jurkat cells treated with Tsp5-Toxin3, Tsp6-Toxin3, Tsp9-Toxin3, and Free Toxinl .
- FIG. 47 shows a plot of in vitro cell proliferation assay results with HER2 antigen positive HCC1954 cells treated with Tspl0-Toxin3, Tspl l-Toxin3, TVLCysl-Toxin3, and Free Toxinl .
- FIG. 48 shows a plot of in vitro cell proliferation assay results with HER2 antigen negative Jurkat cells treated with Tspl0-Toxin3, Tspl l-Toxin3, TVLCysl-Toxin3, and Free Toxinl .
- FIG. 49 shows a plot of in vitro cell proliferation assay results with HER2 antigen positive N87 cells treated with Tspl0-Toxin3, Tspl l-Toxin3, and TVLCysl-Toxin3.
- FIG. 50 shows a plot of in vitro cell proliferation assay results with HER2 antigen positive N87 cells treated with TsplO-Toxinl and Tspl0-Toxin4.
- FIG. 51 shows a plot of in vitro cell proliferation assay results with HER2 antigen negative Jurkat cells treated with TsplO-Toxinl, and Tspl0-Toxin4.
- FIG. 52 shows a plot of in vitro cell proliferation assay results with HER2 antigen positive N87 cells treated with Tsp5-Toxin3, Tsp6-Toxin3, and Tsp9-Toxin3.
- FIG. 53 shows a plot of in vitro cell proliferation assay results with HER2 antigen positive N87 cells treated with Tsp2-Toxin3, Tsp3-Toxin3, and Tsp4-Toxin3.
- FIG. 54 shows a plot of in vitro cell proliferation assay results with CD30 antigen positive Karpas 299 cells treated with Brentuximab, and BsplO.
- FIG. 55 shows a plot of in vitro cell proliferation assay results with CD30 antigen positive Karpas 299 cells treated with Bspl0-Toxin5.
- FIG. 56 shows a plot of in vitro cell proliferation assay results with CD30 antigen positive Karpas 299 cells treated with Bspl0-Toxin3 and Bspl0-Toxin4.
- FIG. 57 shows a plot of in vitro cell proliferation assay results with CD30 antigen positive Karpas 299 cells treated with Bspl0-Toxin6.
- FIG. 58 shows a plot of in vitro cell proliferation assay results with CD30 antigen positive Karpas 299 cells treated with BsplO- MCvcPABC-MMAE.
- FIG. 59 is a gel image showing non-reducing denaturing polyacrylamide gel
- PAGE electrophoresis of trastuzumab light chain extension variants after purification on immobilized protein A.
- lanes 1-12 molecular size marker; TSp2; TSp3; TSp4; TSp5; TSp6; TSp9; TSplO; TSpl 1; TVLCysl; TVLCys2; TVLCys4.
- the size marker ladder in lane 1 indicates the intact proteins are about 150 kDa.
- FIG. 61 is a gel image showing non-reducing denaturing PAGE of trastuzumab light chain extension antibody drug conjugates.
- lanes 1-12 molecular size marker; Tsp2- Toxin3 (DAR 1.92); Tsp3-Toxin3 (DAR 1.88); Tsp4-Toxin3 (DAR 2.06); Tsp5-Toxin3 (DAR 1.46); Tsp6-Toxin3 (DAR 1.80); Tsp9-Toxin3 (DAR 1.32); Tspl0-Toxin3 (DAR 2.12); TsplO- Toxin4 (DAR 1.66); TsplO-Toxin 1 (DAR 2.04); Tspl l-Toxin3 (DAR 2.02); TVLCysl-Toxin3 (DAR 1.06).
- FIG. 63 provides stability data for trastuzumab light chain extension antibody drug conjugates as determined using a thermal stability assay.
- the present invention derives in part from the surprising finding that a C-terminal amino acid extension (also referred to herein as a "payload adaptor") covalently linked to the C-terminus of an antibody light chain as an extension thereof can provide a stable point of attachment for payload, resulting in antibody payload conjugates that are stable and retain affinity for antigen.
- a C-terminal amino acid extension also referred to herein as a "payload adaptor”
- the present disclosure provides antibody light chain polypeptides that include a C-terminal amino acid extension, as well as antibodies and antibody conjugates containing such modified light chain polypeptides, where the C-terminal extension includes one or more cysteine residues.
- Conjugates that include an antibody of the present disclosure conjugated to an agent via a cysteine residue of the C-terminal amino acid extension are also provided.
- the present disclosure further provides nucleic acids encoding an antibody light chain polypeptide that includes a C-terminal amino acid extension including a cysteine residue.
- compositions including the antibodies or conjugates of the present disclosure are also provided, as are methods of making and use of the modified antibodies and conjugates of the present disclosure.
- antibody and “immunoglobulin” include antibodies or immunoglobulins of any isotype, whole antibodies (e.g., antibodies composed of a tetramer which in turn is composed of two heterodimers of a heavy and light chain polypeptide, including whole IgG, IgA, IgD, IgE, or IgM antibodies); half antibodies (e.g., antibodies that include a single dimer of a heavy and light chain polypeptide); antibody fragments (e.g., fragments of whole antibodies, such as fragments of IgG, IgA, IgD, IgE, or IgM antibodies) which retain specific binding to an antigen of interest, including, but not limited to Fab, F(ab')2, Fab', Fv, scFv, bispecific antibodies and diabodies; chimeric antibodies; monoclonal antibodies; humanized antibodies (e.g., humanized monoclonal antibodies, or humanized antibody fragments); or fully human antibodies (an antibody that comprises
- Fv comprises the minimum antibody fragment which contains a complete antigen- recognition and antigen-binding site. This region consists of a dimer of one heavy- and one light- chain variable domain in tight, non-covalent association. It is in this configuration that the three CDRs of each variable domain interact to define an antigen-binding site on the surface of the V H - V L dimer. Collectively, the six CDRs confer antigen-binding specificity to the antibody.
- variable domain or half of an Fv comprising only three CDRs specific for an antigen
- the "Fab” fragment also contains the constant domain of the light chain and the first constant domain (CHi) of the heavy chain.
- Fab fragments differ from Fab' fragments by the addition of a few residues at the carboxyl terminus of the heavy chain CHi domain including one or more cysteines from the antibody hinge region.
- Fab'-SH is the designation herein for Fab' in which the cysteine residue(s) of the constant domains bear a free thiol group.
- F(ab') 2 antibody fragments originally were produced as pairs of Fab' fragments which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.
- Single-chain Fv or “sFv” antibody fragments comprise the V H and V L domains of an antibody, where these domains are present in a single polypeptide chain.
- the Fv polypeptide further comprises a polypeptide linker between the V H and V L domains, which enables the sFv to form the desired structure for antigen binding.
- diabodies refers to small antibody fragments with two antigen-binding sites, which fragments comprise a heavy-chain variable domain (V H ) connected to a light-chain variable domain (V L ) in the same polypeptide chain (V H -V L ).
- V H heavy-chain variable domain
- V L light-chain variable domain
- a “light chain polypeptide” as used herein refers to a polypeptide having at least an antibody light chain variable region (V L ).
- a light chain polypeptide may include a partial or full- length antibody light chain constant region (C L ).
- a "full-length light chain polypeptide” includes a full-length light chain variable region (V L ) and a full-length light chain constant region (C L ).
- the light chain polypeptide may be from any vertebrate species (e.g., mammalian, e.g., human, rodent, and the like).
- a “heavy chain polypeptide” or “heavy chain” as used herein refers to a polypeptide having at least an antibody heavy chain variable region (V H ).
- a heavy chain polypeptide may include a partial or full-length antibody heavy chain constant region (C H ) comprising CHI, CH2 and CH3 domains.
- a "full-length heavy chain polypeptide” includes a full-length heavy chain variable region (V L ) and a full-length heavy chain constant region (C H ).
- Encompassed are heavy chain polypeptides of antibodies (immunoglobulins) from any vertebrate species (e.g., mammalian, e.g., rodent, human, and the like), and any class of immunoglobulin.
- Heavy chain polypeptides and antibodies containing such heavy chain polypeptides, are categorized into classes based on the amino acid sequence of the constant domain of the heavy chain polypeptide. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2.
- immunoglobulins There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2.
- recombinant antibody as used herein is intended to include all antibodies that are prepared, expressed, created, or isolated by recombinant means, such as (i) antibodies expressed using a recombinant expression vector transfected into a host cell; (ii) antibodies isolated from a recombinant, combinatorial antibody library; (iii) antibodies isolated from an animal (e.g. a mouse) that is transgenic for human immunoglobulin genes; or (iv) antibodies prepared, expressed, created, or isolated by any other means that involves splicing of human immunoglobulin gene sequences to other DNA sequences, including, for example, in-vitro translation technology (see, e.g., Yin et al.
- Such recombinant antibodies include humanized, CDR grafted, chimeric, deimmunized, and in vitro generated antibodies; and can optionally include constant regions derived from human germline immunoglobulin sequences.
- humanized antibody refers to immunoglobulins, half antibodies,
- immunoglobulin chains e.g., a light chain polypeptide
- fragments thereof such as Fv, scFv, Fab, Fab', F(ab')2 or other antigen-binding subsequences of antibodies
- the humanized antibodies may be human immunoglobulins (recipient antibody) in which residues from a complementary determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat, lama, camel or rabbit having the desired specificity, affinity and capacity.
- CDR complementary determining region
- donor antibody such as mouse, rat, lama, camel or rabbit having the desired specificity, affinity and capacity.
- Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues.
- a humanized antibody may comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences.
- treating is meant alleviating or abrogating a disease or disorder and/or at least one of its attendant symptoms.
- to “alleviate” a disease or disorder means reducing the severity and/or occurrence frequency of the symptoms of the disease or disorder. It will be understood that references herein to “treating,” “treat,” or
- treatment include references to curative, palliative and prophylactic treatment.
- terapéuticaally effective amount or “efficacious amount” is meant a dosage sufficient to produce a desired result, e.g., an amount sufficient to effect beneficial or desired therapeutic (including preventative) results, such as a reduction in a symptom of a disease (e.g., cancer or any other disease of interest), as compared to a control.
- the “therapeutically effective amount” will vary depending on the antibody or conjugate, the disease and its severity, and the age, weight, etc., of the patient to be treated.
- the present disclosure provides payload adaptors (also referred to herein as a "C-terminal amino acid extension” or “C-terminal extension”) for the attachment of payloads to antibodies, as well as antibodies comprising such payload adaptors.
- payload adaptors also referred to herein as a "C-terminal amino acid extension” or “C-terminal extension” for the attachment of payloads to antibodies, as well as antibodies comprising such payload adaptors.
- the payload adaptors of the present disclosure are protein modules that serve as substrates for covalent attachment of payloads, with each payload adaptor constituting a C- terminus extension of an antibody light chain and thereby linking one or more payloads to an antibody.
- the payload adaptors comprise at least one cysteine residue for payload attachment.
- Payload adaptors of the present disclosure are capable of being expressed as C-terminus extensions of antibody light chains and are capable of covalent conjugation to a wide variety of payloads with the use of appropriate chemistry such that the antibodies comprising payloads and payload adaptors exhibit stability and retain affinity for antigen.
- an antibody comprising a payload adaptor of the present disclosures does not comprise a cysteine substitution within its native antibody sequence, which might otherwise be introduced to provide a compensatory disulfide bond accommodating addition of a polypeptide to the C-terminus end of a light chain.
- an antibody comprising a payload adaptor of the present disclosures contains all cysteine residues that are present in the parent antibody.
- the payload adaptor comprises multiple cysteine residues that do not form an intramolecular disulfide bond or a disulfide bond with another payload adaptor.
- a payload adaptor does not specifically bind antigen. In one embodiment, the payload adaptor does not contribute an epitope binding activity to the antibodies or antibody conjugates of the invention. In this embodiment, antigen binding by the antibodies and antibody conjugates of the present disclosure is determined by elements other than the payload adaptors. In one embodiment, a payload adaptor does not contain a ligand binding domain of a growth factor receptor, such as an EGF receptor). In another embodiment, the payload adaptor does not contain a ligand of a growth factor receptor (e.g., does not contain a ligand of an EGF receptor).
- the present disclosure provides light chain polypeptides having a C-terminal extension having one or more cysteine residues, and antibodies having at least one of such modified light chain polypeptides.
- the light chain polypeptide having the C-terminal extension can contain an amino acid sequence of a light chain polypeptide of any type (e.g., a lambda ( ⁇ ) or kappa ( ⁇ ) light chain polypeptide) and can contain amino acid sequences of a light chain polypeptide of any origin of interest, e.g., any vertebrate species (e.g., mammalian, e.g., rodent, human, and the like).
- C-terminal light chain polypeptide extension refers to an amino acid (e.g., a cysteine) or a contiguous stretch of two or more amino acids located C-terminal to the residue of the light chain polypeptide that would otherwise constitute the C-terminal residue in a parental light chain polypeptide absent the extension.
- amino acid e.g., a cysteine
- the parental light chain polypeptide only includes a light chain variable region (V L ) (e.g., the parental antibody may be an ScFv), such that the extension is C-terminal to (e.g., extends from) the residue that would otherwise constitute the C-terminus of a V L in a parental light chain polypeptide.
- the parental light chain polypeptide includes a light chain variable region (V L ) and a partial light chain constant region (C L ), such that the extension is C-terminal to (e.g., extends from) the residue that would otherwise constitute the C- terminus of a partial C L in a parental light chain polypeptide.
- the parental light chain polypeptide is a full-length light chain polypeptide that includes a full-length light chain variable region (V L ) and a full- length light chain constant region (C L ), such that the extension is C-terminal to (e.g., extends from) the residue that would otherwise constitute the C-terminus of a full-length C L in a parental light chain polypeptide.
- the N-terminal portion of the extension includes at least a portion of a sequence that would otherwise be present in a full-length parental light chain polypeptide, such that extending the C-terminus of the parental light chain polypeptide includes "adding back" parental sequence as part of the "extension.”
- the parental light chain polypeptide includes a deletion of the terminal cysteine normally present at the C-terminus of a full-length wild-type light chain polypeptide, such that the light chain extension is C-terminal to (e.g., extends from) the residue immediately N-terminal to the position in which the C-terminal cysteine is deleted.
- the parental light chain polypeptide includes a substitution of the terminal cysteine normally present at the C-terminus of a full-length wild-type light chain polypeptide.
- the parental antibody has a truncated heavy chain polypeptide, e.g., a heavy chain polypeptide that only includes a heavy chain variable region (V H ), or a heavy chain polypeptide that includes a heavy chain variable region (V H ) and a portion of heavy chain constant region (C H ).
- the C-terminal light chain polypeptide extension may comprise native (e.g., wild-type) light chain polypeptide sequence unpaired with heavy chain polypeptide sequence (due to the truncation).
- such a C-terminal light chain polypeptide extension may further include one or more non-native amino acids (e.g., one or more cysteines not present in the parental light chain polypeptide), which in certain aspects may be a non-native sequence of two or more amino acids.
- one or more non-native amino acids e.g., one or more cysteines not present in the parental light chain polypeptide
- the present disclosure provides an antibody including a light chain polypeptide that includes a C-terminal amino acid extension including a cysteine residue.
- the present disclosure provides an antibody that includes a light chain polypeptide including a C-terminal amino acid extension that includes a cysteine residue, which C-terminal amino acid extension does not specifically bind antigen (e.g., the extension does not include an antigen-binding portion of an antibody or an antigen-binding portion of an antibody fragment).
- the present disclosure provides an antibody that includes at least one monoepitopic antigen-binding dimer, where the monoepitopic antigen-binding dimer includes a heavy chain polypeptide and a light chain polypeptide that includes a C-terminal amino acid extension, which extension includes a cysteine residue.
- the monoepitopic antigen-binding dimer includes a heavy chain polypeptide and a light chain polypeptide that includes a C-terminal amino acid extension, which extension includes a cysteine residue.
- Each of the two monoepitopic dimers may bind to the same epitope. In other aspects, each of the two monoepitopic dimers binds a different epitope .
- “Monoepitopic antigen-binding domain” indicates an antigen-binding domain formed by interaction of the CDRs of a heavy chain polypeptide and the CDRs of a light chain polypeptide.
- Monoepitopic antigen-binding domains can be defined by, for example, a dimer comprising a heavy chain polypeptide and a light chain polypeptide, or, in the case of a single chain antibody (ScFv) a monomeric fusion protein comprising a heavy chain polypeptide and a light chain polypeptide.
- ScFv single chain antibody
- the amino acid extension of the light chain polypeptide does not specifically bind antigen.
- Antibodies of the present disclosure include antibodies comprising the same or different monoepitopic antigen-binding dimer.
- an antibody comprising a dimer of heterodimers may include: 1) a first monoepitopic antigen-binding domain comprising a heavy chain polypeptide and a light chain polypeptide, and a second monoepitopic antigen-binding domain comprising a heavy chain polypeptide and a light chain polypeptide, wherein one or both of the light chain polypeptides comprises a C-terminal amino acid extension, and wherein the first and second monoepitopic antigen-binding domains bind the same epitope; or 2) first and second monoepitopic antigen binding domains, wherein one or both of the light chain polypeptides of the domains comprises a C-terminal amino acid extension, where the antigen-binding region of the first monoepitopic antigen-binding domain binds a different epitope as that
- the C-terminal extension includes two or more contiguous cysteines.
- the extension may include two adjacent cysteines having non-cysteine-containing spacer sequences N-terminal and C-terminal to the two adjacent cysteines.
- Having contiguous cysteines (e.g., two adjacent cysteines) in the C-terminal extension finds use, e.g., when the conjugation or labeling strategy includes metal chelation, when the conjugation or labeling strategy involves "bridging" (e.g., as is the case with certain dihalo- maleimide conjugation chemistries, and the like).
- the present disclosure provides conjugates and methods of making the same in which the agent (e.g., a drug or labeling agent) is attached to multiple contiguous cysteines (e.g., 2 adjacent cysteines), either directly or through one or more linkers.
- agent e.g., a drug or labeling agent
- multiple contiguous cysteines e.g., 2 adjacent cysteines
- the C-terminal extension includes an N-terminal cysteine that when taken together with the parental light chain terminal cysteine provides two contiguous cysteines that find use as described above.
- the present disclosure provides conjugates and methods of making the same in which the agent (e.g., a drug or labeling agent) is attached to multiple non-contiguous cysteines, either directly or through one or more linkers.
- the cysteines of the C-terminal extension are separated by one or more spacers, such that the cysteines are not contiguous residues of the C-terminal extension.
- spacer is meant one or more consecutive non-cysteine amino acids disposed between two cysteine residues in the extension; between what would otherwise constitute the C-terminal residue of the light chain polypeptide, or fragment thereof containing a light chain variable region and at least a portion of a light chain constant region, and the first cysteine residue in the C-terminal extension; and/or optionally one or more consecutive non-cysteine amino acids disposed C-terminal to the most C-terminal cysteine in the extension. Any number of spacers may be provided in the C-terminal extension.
- the C-terminal extension can include from 1 to 50 spacers, from 1 to 40 spacers, from 1 to 30 spacers, from 1 to 20 spacers, from 1 to 10 spacers (e.g., from 2 to 10 spacers), or 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 spacers.
- each of the spacers may have the same amino acid sequence.
- the amino acid sequence of at least two of the spacers may be different.
- a cysteine may be present between each of the spacers.
- at least two of the spacers are contiguous, e.g., the spacers are not separated by one or more cysteine residues.
- the spacer may include any of the 20 non-cysteine, naturally-occurring, genetically encodable amino acids (alanine (A), arginine (R), asparagine (N), aspartic acid (D), glutamic acid (E), glutamine (Q), glycine (G), histidine (H), isoleucine (I), leucine (L), lysine (K), methionine (M), phenylalanine (F), proline (P), serine (S), threonine (T), tryptophan (W), tyrosine (Y), and/or valine (V)), or variants thereof (e.g., variants that arise as a result of post- translation modification), naturally occurring non-encodable or non-natural amino acids, and may be of any desired sequence and length.
- the spacer includes from 1 to 30 amino acids, such as from 3 to 20 amino acids, 3 to 15 amino acids, 3 to 10 amino acids, 3 to 5 amino acids, and may be, e.g., from 4 to 17 amino acids.
- the spacer may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more amino acids, and may in some instances contain not more than 30 or more than 25 amino acids, may be of any desired amino acid sequence with the proviso the spacer does not include a cysteine residue.
- the spacer includes at least one glycine (G) residue and at least one serine (S) residue.
- the spacer may contain one or more glycine residues and one or more serine residues.
- a spacer of interest is a spacer having the sequence GGGS (SEQ ID NO: l).
- the spacer may include or consist of any of the following amino acid sequences: AKTTPKLEEGEFSEAR (SEQ ID NO:2); AKTTPKLEEGEFSEARV (SEQ ID NO:3); AKTTPKLGG (SEQ ID NO:4); SAKTTPKLGG (SEQ ID NO:5);
- AKTTPKLEEGEFSEARV SEQ ID NO:6; SAKTTP (SEQ ID NO:7); SAKTTPKLGG (SEQ ID NO:8); RADAAP (SEQ ID NO:9); RADAAPTVS (SEQ ID NO: 10); RADAAAAGGPGS (SEQ ID NO: l 1); RADAAAA(G 4 S) 4 (SEQ ID NO: 12), SAKTTP (SEQ ID NO: 13);
- SAKTTPKLGG SEQ ID NO: 14
- S AKTTPKLEEGEFSEARV SEQ ID NO: 15
- ADAAP SEQ ID NO: 16
- ADAAPTVSIFPP SEQ ID NO: 17
- TVAAP SEQ ID NO: 18
- TVAAPSVFIFPP SEQ ID NO: 19
- QPKAAP SEQ ID NO:20
- QPKAAP S VTLFPP SEQ ID NO:21
- AKTTPP SEQ ID NO:22
- AKTTPPSVTPLAP SEQ ID NO:23
- AKTTAP SEQ ID NO:24
- AKTTAP S VYPL AP SEQ ID NO:25
- ASTKGP SEQ ID NO:26
- ASTKGPSVFPLAP SEQ ID NO:27
- GGGGSGGGGSGGGGS SEQ ID NO:28
- GHEAAAVMQVQYPAS (SEQ ID NO:31); AA; GGGGS (SEQ ID NO:128);
- GGGGSSGGGGSS (SEQ ID NO: 131); or variants thereof that include 1, 2, 3, 4, or 5 amino acid substitutions.
- the C-terminal extension of the light chain polypeptide includes an amino acid sequence having endogenous human amino acid sequences or modified human amino acid sequences. These may include human antibody hinge region sequences, T-cell receptor sequences, or any other human protein sequences of interest. Additional amino acid sequences that may be employed include, but are not limited to, extracellular protein amino acid sequences, as well as the sequences of extracellular domains of proteins present on a cell surface.
- the extension includes an endogenous human amino acid sequence that includes one or more naturally occurring cysteine residues.
- the non-cysteine- containing amino acid sequences N-terminal and C-terminal to the cysteine residues constitute spacer sequences of the C-terminal extension.
- the extension includes a modified human amino acid sequence in which one or more cysteines has been introduced into the endogenous human amino acid sequence by insertion or substitution.
- Naturally occurring cysteine residues may also be substituted or deleted in the context of such sequences.
- the immunogenicity of the antibody (or conjugate thereof) when administered to a patient is reduced as compared to a corresponding extension lacking the human amino acid sequence or modified human amino acid sequence.
- Preferred spacers include amino acid sequences with at least 85%, 90%, 95%, 98% or 100%) sequence identity to the wild-type sequence, such as a portion of a hinge region, or fragment thereof, of a wild-type IgM, IgG, IgA, IgE or IgD antibody molecule or T cell receptor.
- the "hinge" region refers to the amino acid sequence of an antibody (such as depicted in the examples of FIGs. 1 and 2) located between the C H I and C H 2 domains of a heavy chain polypeptide, e.g., a heavy chain polypeptide of an IgG, IgA or IgD antibody.
- the hinge region of the constructs of the present disclosure may vary in length and amino acid sequence.
- the hinge regions of human IgGi, IgG 2 and IgG 4 are 12-15 amino acids in length, while human IgG 3 has a 62 amino acid hinge region.
- Human IgD antibody molecules have a 64 amino acid hinge region.
- the immunogenicity of the antibody (or conjugate thereof) when administered to a patient is reduced as compared to a corresponding extension lacking the hinge region sequence, and the flexibility of the extension may be increased relative to an extension lacking the hinge region sequence.
- Non-limiting examples of hinge region amino acid sequences include but are not limited to, ESSCDVKLVEKSFET (SEQ ID NO: 32) (T cell receptor alpha constant); DCGFTS (SEQ ID NO: 33) (T cell receptor beta constant); DVITMDPKDNCSKDAN (SEQ ID NO: 34) (T cell receptor gamma constant);
- DH VKPKETENTKQP SKS CHKPK (SEQ ID NO: 35) (T cell receptor delta constant); EPKSCDKTHTCPPCP (SEQ ID NO: 36) (IgCHGl); ERKCCVECPPCP (SEQ ID NO: 37) (IgCHG2); ELKTPLGDTTHTCPRCP (SEQ ID NO: 38) (IgCH3-Hl); EPKSCDTPPPCPRCP (SEQ ID NO: 39) (IgCH3-H2, IgCH3-H3, and IgCH3-H4); ESKYGPPCPSCP (SEQ ID NO: 40) (IgH4); VPPPPP (SEQ ID NO: 41) (IgA2) and
- Non-limiting examples of non-cysteine-containing amino acid sequences derived from hinge region amino acid sequences, of which all or a portion thereof (e.g., at least 2, 3, 4, 5, 6 or more contiguous residues) may be used as spacers in the C-terminal extensions of the present disclosure include but are not limited to, ESS (SEQ ID NO: 43); DVKLVEKSFET (SEQ ID NO: 44); GFTS (SEQ ID NO: 45); DVITMDPKDN (SEQ ID NO: 46); SKDAN (SEQ ID NO: 47); DH VKPKETENTKQP SKS (SEQ ID NO: 48); HKPK (SEQ ID NO: 49); EPKS (SEQ ID NO: 50); DKTHT (SEQ ID NO: 51); ERK (SEQ ID NO: 52); ELKTPLGDTTHT (SEQ ID NO: 53); DTPPP (SEQ ID NO: 54); VE (SEQ ID NO: 55); PR (SEQ ID NO: 56); PP
- the C-terminal extension of a light chain polypeptide of the present disclosure may be designed to include any desired combination of one or more spacers (or no spacer) and one or more cysteine residues.
- an aspect of the present disclosure is to provide an extension at the C-terminus of a light chain polypeptide having one or more cysteines (e.g., spaced apart from each other or not spaced apart from each other; spaced from the C-terminus of the parental light chain polypeptide or not spaced from the C-terminus of the parental light chain polypeptide; and/or spaced from the C-terminus of the light chain extension or not spaced from the C- terminus of the light chain extension), so that one may control the corresponding number and spacing of agents (e.g., cytotoxic agents, labeling agents, and/or the like) linked to such cysteine(s) in a conjugated product of such an antibody.
- the C-terminal extension comprises an amino acid sequence which may be represented, from N-terminus to C
- X and X' represent a spacer of one or more amino acids, wherein the amino acid sequence of each X and X' is independently selected from any amino acid sequence of interest, including any of the examples of spacer amino acid sequences provided herein;
- C represents a cysteine residue
- a, b, c, d, e and f are integers independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20, wherein the sum of b and e is at least 1, and the sum of c and f is at least 1.
- X and X' may be the same or different. Where c is greater than 1 , then each X of (X a Cb) c may be the same or different amino acid sequence within each repeat unit of (X a Cb) c . Where d is greater than 1, then each X' of (X'dC e ) f may the same or different amino acid sequence within each repeat unit of (X' d C e ) f .
- the present disclosure also provides nucleic acids encoding a C-terminal extension of Formula I, as well as nucleic acids encoding a light chain polypeptide comprising a C-terminal extension of Formula I.
- the C-terminal extension may be represented, from N-terminus to C-terminus, by Formula I above, where b and e are integers independently selected from 0, 1, and 2, wherein the sum of b and e is at least 1, and a, c, d, and f are integers independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20, where the sum of c and f is at least 1.
- the C-terminal extension may be represented, from N-terminus to C-terminus, by Formula I above, where b and e are integers independently selected from 0, 1, and 2, wherein the sum of b and e is at least 2, and a, c, d, and f are integers independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20, where the sum of c and f is at least 1.
- the C-terminal extension may be represented, from N-terminus to C-terminus, by Formula I above, where b and e are integers independently selected from 0, 1, and 2, where the sum of b and e is at least 1, a and d are integers independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10, and c and f are integers independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10, where the sum of c and f is at least 1.
- the C-terminal extension may be represented, from N-terminus to C-terminus, by the formula I above, wherein b and e are integers independently selected from 0, 1, and 2, wherein the sum of b and e is at least 2, a and d are integers independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10, and c and f are integers independently selected from 0, 1,
- the C- terminal extension has the sequence GGGSC (SEQ ID NO:60) (also referred to herein as "Cysl").
- GGGSC SEQ ID NO:60
- FIG. 1 An example of an antibody having a light chain polypeptide with a C-terminal extension according to this embodiment is schematically illustrated in FIG. 1.
- antibody 100 includes two light chain polypeptides that include light chain variable (V L ) and constant (C L ) domains, and C-terminal extensions 102 and 104 having the sequence GGGSC extending from the C-terminal residue of each of the (C L ) domains.
- X is GGGS (SEQ ID NO:l)
- a is 1
- c is 1
- f is 0.
- the C-terminal extension has the sequence GGGSCC (SEQ ID NO:61).
- the C-terminal extension has the sequence GGGSGGGSC (SEQ ID NO:62) when each spacer sequence is the same.
- the C-terminal extension has the sequence GGGSCGGGSC (SEQ ID NO:63) (also referred to herein as "Cys2”) when each spacer sequence is the same.
- GGGSCAKTTPKLEEGEFSEARC SEQ ID NO:89.
- GGGSCAKTTPKLEEGEFSEARCAKTTPKLEEGEFSEARC SEQ ID NO:90.
- the C-terminal extension has the sequence AKTTPKLEEGEF SE ARAKTTPKLEEGEF SE ARC (SEQ ID NO:69) when each spacer sequence is the same.
- the C- terminal extension has the sequence AKTTPKLEEGEF SE ARC AKTTPKLEEGEF SE ARC (SEQ ID NO: 70) when each spacer sequence is the same.
- AKTTPKLEEGEFSEARCAKTTPKLEEGEFSEARCAKTTPKLEEGEFSEARCAKTTPKLEEGEFSEARCAKTTPKLEE GEFSEARC (SEQ ID NO:71) when each spacer sequence is the same.
- AKTTPKLEEGEFSEARCAKTTPKLEEGEFSEARAKTTPKLEEGEFSEARAKTTPKLEEGEF SEARC (SEQ ID NO:72) when each spacer sequence is the same.
- AKTTPKLEEGEFSEARAKTTPKLEEGEFSEARCAKTTPKLEEGEFSEARC (SEQ ID NO:73) when each spacer sequence is the same.
- AKTTPKLEEGEFSEARCAKTTPKLEEGEFSEARAKTTPKLEEGEFSEARC (SEQ ID NO:74) when each spacer sequence is the same.
- a C-terminal extension of the present disclosure may be represented by Formula II:
- X, X', and X" represent a spacer of one or more amino acids, wherein the amino acid sequence of each X, X' and X' ' is independently selected from any amino acid sequence of interest, including any of the examples of spacer amino acid sequences provided herein;
- C is a cysteine residue:
- a, b, c, d, e, f, g, h and i are integers independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20, where the sum of b, e and h is at least 1, and the sum of c, f and i is at least 1.
- X, X', and X' ' may be the same or different. Where c is greater than 1 , then each X of (X a Cb) c may be the same or different amino acid sequence within each repeat unit of (X a C b ) c .
- each X' of (X' d C e ) f may the same or different amino acid sequence within each repeat unit of (X'dC e ) f .
- each X" of (X" g C h )i may the same or different amino acid sequence within each repeat unit
- a C-terminal extension of the present disclosure may include (e.g., consist of) an amino acid sequence selected from: EPKSCDKTHTC (SEQ ID NO:92) (also referred to herein as extension 1); EPKSCDKTHTCPPC (SEQ ID NO:93) (also referred to herein as extension 2); EPKSC (SEQ ID NO:94) (also referred to herein as extension 3); ESKYGPPC (SEQ ID NO:95) (also referred to herein as extension 4); ERKCCVECPPC (SEQ ID NO: 96) (also referred to herein as extension 5); ERKC (SEQ ID NO:97) (also referred to herein as extension 6);
- DVITMDPKDNC (SEQ ID NO:98) (also referred to herein as extension 7);
- DHVKPKETENTKQPSKSCHKPK (SEQ ID NO:99) (also referred to herein as extension 8); ESSC (SEQ ID NO: 100) (also referred to herein as extension 9); ESSCDVKLV (SEQ ID NO: 101) (also referred to herein as extension 10); DHVKPKETENTKQPSKSC (SEQ ID NO: 102) (also referred to herein as extension 11); DVITMDPKDNCSKDAN (SEQ ID NO: 103) (also referred to herein as extension 12); CAA, CCAA (SEQ ID NO: 128), AACAA (SEQ ID NO: 129), and GGGGSCAA (SEQ ID NO:130).
- the present disclosure also provides nucleic acids encoding any of the C-terminal extensions described herein (e.g., C-terminal extension of Formula II, etc.), as well as nucleic acids encoding any of the light chain polypeptides described herein (e.g., light chain
- polypeptides comprising a C-terminal extension of Formula II, etc.
- the present disclosure provides antibodies having at least one light chain polypeptide having a C-terminal extension of the present disclosure.
- the antibody has two light chain polypeptides having a C-terminal extension of the present disclosure.
- such antibodies can be conjugated to an agent/payload (e.g., a drug) through covalent attachment to at least one cysteine in the C-terminal extension present in at least one of the light chain polypeptides of the antibody.
- the payload is directly attached to the cysteine of the C-terminal extension.
- the payload is attached via a linker to the cysteine of the C-terminal extension.
- the payload is preferentially attached to the cysteine of the C-terminal extension over a cysteine outside the C- terminal amino acid extension.
- the payload is exclusively attached to the cysteine of the C-terminal extension.
- the present disclosure provides nucleic acids encoding a light chain polypeptide having a C-terminal amino acid extension (referred to herein for convenience as a "modified light chain polypeptide"), as well as vectors and host cells containing such nucleic acids.
- the modified light chain polypeptide encoded by the nucleic acid may include any of the features described above, in any combination.
- the C-terminal extension portion of the light chain polypeptide may include any of the C-terminal extension features described above with respect to the length of the extension, the amino acid sequence of the extension, the number of spacers in the extension and amino acid sequences thereof, extension configurations based on combinations of one or more spacers and one or more cysteine residues, and any other aspects of the C-terminal extensions described above and elsewhere herein.
- a nucleic acid e.g., DNA or RNA
- encoding a C-terminal extension-containing light chain polypeptide can be operably linked to one or more regulatory elements, such as a promoter and enhancer, that allow expression of the nucleic acid in a host cell (e.g., a cell that is genetically modified to synthesize the encoded modified light chain polypeptide).
- promoters include, but are not limited to, a bacteriophage T7 RNA polymerase promoter; a trp promoter; a lac operon promoter; a hybrid promoter, e.g., a lac/tac hybrid promoter, a tac/trc hybrid promoter, a trp/lac promoter, a T7/lac promoter; a trc promoter; a tac promoter, and the like; an araBAD promoter; in vivo regulated promoters, such as an ssaG promoter or a related promoter, a pagC promoter; a nirB promoter; a sigma70 promoter, e.g., a consensus sigma70 promoter; a stationary phase promoter, e.g., a dps promoter, an spv promoter, and the like; a promoter
- Examples of strong promoters for use in prokaryotes such as Escherichia coli include, but are not limited to Trc, Tac, T5, T7, and PLambda.
- operators for use in bacterial host cells include a lactose promoter operator (Lacl repressor protein changes conformation when contacted with lactose, thereby preventing the Lacl repressor protein from binding to the operator), a tryptophan promoter operator (when complexed with tryptophan, TrpR repressor protein has a conformation that binds the operator; in the absence of tryptophan, the TrpR repressor protein has a conformation that does not bind to the operator), and a tac promoter operator.
- the promoter can be a constitutive promoter such as an ADHl promoter, a PGKl promoter, an ENO promoter, a PYKl promoter and the like; or a regulatable promoter such as a GAL1 promoter, a GAL 10 promoter, an ADH2 promoter, a PH05 promoter, a CUP1 promoter, a GAL7 promoter, a MET25 promoter, a MET3 promoter, a CYC1 promoter, a HIS3 promoter, an ADHl promoter, a PGK promoter, a GAPDH promoter, an ADC1 promoter, a TRP1 promoter, a URA3 promoter, a LEU2 promoter, an ENO promoter, a TP1 promoter, and AOX1 (e.g., for use in Pichia).
- a constitutive promoter such as an ADHl promoter, a PGKl promoter, an ENO promoter,
- a nucleotide sequence encoding the modified light chain polypeptide can be present in an expression vector and/or a cloning vector.
- the corresponding nucleotide sequences encoding the two or more polypeptides may be cloned in the same or separate vectors.
- An expression vector can include a selectable marker, an origin of replication, and other features that provide for replication and/or maintenance of the vector.
- Expression vectors generally have convenient restriction sites located near the promoter sequence to provide for the insertion of nucleic acid sequences encoding heterologous proteins.
- expression vectors include, but are not limited to, viral vectors (e.g. viral vectors based on vaccinia virus; poliovirus; adenovirus; adeno-associated virus; SV40; herpes simplex virus; human immunodeficiency virus; a retroviral vector (e.g., Murine Leukemia Virus, spleen necrosis virus, and vectors derived from retroviruses such as Rous Sarcoma Virus, Harvey Sarcoma Virus, avian leukosis virus, human immunodeficiency virus, myeloproliferative sarcoma virus, and mammary tumor virus); and the like.
- viral vectors e.g. viral vectors based on vaccinia virus; poliovirus; adenovirus; adeno-associated virus; SV40; herpes simplex
- a host cell that includes any of the nucleic acids encoding the modified light chain polypeptide, or vectors including the same.
- the host cell is a prokaryotic host cell or a eukaryotic host cell.
- the host cell may be an isolated genetically modified host cell (e.g., an in vitro cell) that is genetically modified (e.g., transformed or transfected) with a nucleic acid of the present disclosure.
- a genetically modified host cell of the present disclosure can produce a modified light chain polypeptide of the present disclosure, which antibody light chain polypeptide can have any of the features described elsewhere herein.
- host cells include eukaryotic host cells, such as a mammalian cell, an insect host cell, a yeast cell; and prokaryotic cells, such as a bacterial cell.
- eukaryotic host cells such as a mammalian cell, an insect host cell, a yeast cell
- prokaryotic cells such as a bacterial cell.
- Introduction of a the nucleic acid into the host cell can be effected, for example by calcium phosphate precipitation, DEAE dextran mediated transfection, liposome-mediated transfection, electroporation, or other known methods.
- mammalian host cells include primary cells and immortalized cell lines.
- Suitable mammalian cell lines include human cell lines, non-human primate cell lines, rodent (e.g., mouse, rat) cell lines, and the like.
- Suitable mammalian cell lines include, but are not limited to, HeLa cells (e.g., American Type Culture Collection (ATCC) No. CCL-2), CHO cells (e.g., ATCC Nos. CRL9618, CCL61, CRL9096), 293 cells (e.g., ATCC No. CRL-1573), Vero cells, NIH 3T3 cells (e.g., ATCC No. CRL-1658), Huh-7 cells, BHK cells (e.g., ATCC No.
- CCL10 PC12 cells
- COS cells COS-7 cells
- RATI cells mouse L cells
- HEK cells ATCC No. CRL1573
- HLHepG2 cells HLHepG2 cells, and the like.
- yeast host cells include, but are not limited to, Pichia pastoris, Pichia finlandica, Pichia trehalophila, Pichia koclamae, Pichia membranaefaciens, Pichia opuntiae, Pichia thermotolerans, Pichia salictaria, Pichia guercuum, Pichia pijperi, Pichia stiptis, Pichia methanolica, Pichia sp., Saccharomyces cerevisiae, Saccharomyces sp., Hansenula polymorpha, Kluyveromyces sp., Kluyveromyces lactis, Candida albicans, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Trichoderma reesei, Chrysosporium lucknowense, Fusarium sp., Fusarium gramineum, Fusarium gram
- prokaryotic cells include, but are not limited to, any of a variety of laboratory strains of Escherichia coli, Lactobacillus sp., Salmonella sp., Shigella sp., and the like.
- Salmonella strains which can be employed in the present invention include, but are not limited to, Salmonella typhi and S. typhimurium.
- Suitable Shigella strains include, but are not limited to, Shigella flexneri, Shigella sonnei, and Shigella disenteriae.
- the laboratory strain is one that is non-pathogenic.
- Non-limiting examples of other suitable bacteria include, but are not limited to, Bacillus subtilis, Pseudomonas pudita, Pseudomonas aeruginosa, Pseudomonas mevalonii, Rhodobacter sphaeroides, Rhodobacter capsulatus, Rhodospirillum rubrum, Rhodococcus sp., and the like.
- modified light chain polypeptides i.e., a light chain polypeptide having a C-terminal extension of the present disclosure
- antibodies containing one or more of such modified light chain polypeptides are also contemplated.
- a method of making a light chain polypeptide of an antibody including expressing in a host cell a nucleic acid encoding a light chain polypeptide that includes a C-terminal amino acid extension as described herein.
- the light chain polypeptide may be produced by any convenient method, e.g., conventional synthetic methods for protein synthesis, recombinant DNA methods, etc.
- the light chain polypeptide having a C-terminal amino acid extension may be produced in combination with the production of a heavy chain polypeptide of interest, or may be combined with a heavy chain polypeptide following production (e.g., by fusion of recombinant cells which separately express a light chain polypeptide of the present disclosure and a heavy chain polypeptide).
- nucleic acid sequences can encode the desired light chain polypeptide.
- the nucleic acid sequence encoding the light chain polypeptide can be produced by de novo solid-phase DNA synthesis, by polymerase chain reaction (PCR) mutagenesis (e.g., overlapping PCR) of a nucleic acid that encodes a light chain polypeptide of a parental antibody that lacks a C-terminal amino acid extension.
- PCR polymerase chain reaction
- mutagenesis e.g., overlapping PCR
- Suitable expression vectors are typically replicable in the host organisms either as episomes or as an integral part of the host chromosomal DNA. Commonly, expression vectors contain selection markers (e.g., ampicillin-resistance, hygromycin-resistance, tetracycline resistance, kanamycin resistance or neomycin resistance) to permit detection of those cells transformed with the desired DNA sequences. Examples of expression vectors and host cells are discussed above.
- the host cells can be prokaryotic cells, (e.g., Escherichia coli, bacilli (such as Bacillus subtilis) Salmonella, Serratia, Pseudomonas species, and the like), yeast cells (e.g., Saccharomyces (e.g., S. cerevisiae), Pichia and the like), and mammalian cells (e.g., CHO cell lines, Cos cell lines, HeLa cells, myeloma cell lines, transformed B-cells, hybridomas and the like).
- prokaryotic cells e.g., Escherichia coli, bacilli (such as Bacillus subtilis) Salmonella, Serratia, Pseudomonas species, and the like
- yeast cells e.g., Saccharomyces (e.g., S. cerevisiae), Pichia and the like
- mammalian cells e.g., CHO cell lines, Cos cell lines, He
- the synthesis may proceed via liquid-phase or solid-phase.
- Solid phase polypeptide synthesis in which the C- terminal amino acid of the sequence is attached to an insoluble support followed by sequential addition of the remaining amino acids in the sequence, is an example of a suitable method for the chemical synthesis of the light chain polypeptide.
- SPPS Solid phase polypeptide synthesis
- Various forms of SPPS such as Fmoc and Boc, are available for synthesizing the light chain polypeptide.
- Techniques for solid phase synthesis are available in the art. For example, small insoluble, porous beads are treated with functional units on which peptide chains are built. After repeated cycling of
- the free N-terminal amine of a solid-phase attached is coupled to a single N-protected amino acid unit. This unit is then deprotected, revealing a new N-terminal amine to which a further amino acid may be attached.
- the peptide remains immobilized on the solid- phase and undergoes a filtration process before being cleaved off.
- the modified light chain polypeptides can be purified according to standard procedures.
- the modified light chain polypeptide, either alone or as part of an antibody can be substantially pure, e.g., at least about 80% to 85% pure, at least about 85% to 90%> pure, at least about 90%) to 95%o pure, or 98%> to 99%, or more, pure, e.g., free from contaminants such as cell debris, macromolecules other than the light chain polypeptide, and the like.
- the antibody may be a recombinant antibody, e.g., a chimeric, humanized, fully human, bispecific, deimmunized, and/or an in vitro generated antibody.
- the present disclosure provides antibodies having at least one light chain polypeptide having a C-terminal extension, where at least one cysteine of the C-terminal extension is conjugated to an agent (e.g., drug).
- an agent e.g., drug
- the light chain polypeptide having a C-terminal extension of the antibody portion of the conjugates of the present disclosure may include any of the features described above, and in any combination.
- the C-terminal extension of the antibody portion of the conjugate may include any of the C-terminal extension features described above with respect to the length of the extension, the amino acid composition of the extension, the number of spacers in the extension and amino acid sequences thereof, extension configurations based on combinations of one or more spacers and one or more cysteine residues, and any other aspects of the C-terminal extensions described above and elsewhere herein.
- the antibody conjugates of the present disclosure have at least one light chain polypeptide having a C-terminal extension, where at least one cysteine residue of the C- terminal extension is conjugated to an agent.
- each of the light chain polypeptides of the antibody conjugates of the present disclosure have a C-terminal extension, where at least one cysteine residue of the C-terminal extensions of each of the modified light chain polypeptides is conjugated to an agent.
- a first agent may be conjugated to a cysteine residue of the C-terminal extension of a first modified light chain polypeptide
- a second agent may be conjugated to a cysteine residue of the C-terminal extension of a second light chain polypeptide.
- the agent is preferentially attached to the cysteine residue of the C-terminal extension rather than a cysteine residue outside the C-terminal amino acid extension. In certain aspects, the agent is exclusively attached to the cysteine residue of the C-terminal extension.
- Antibody conjugates of the present disclosure can include any number of agents conjugated to the C-terminal extension of the modified light chain polypeptide(s), according to the number of cysteines in the C-terminal extension available for conjugation, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or more cysteines.
- the C-terminal extension of a modified light chain polypeptide(s) of the antibody is conjugated to an agent or agents via covalent binding to at least 1, 2, 3, 4, 5, or more cysteines of the C-terminal extension. In some embodiments the C-terminal extension of a modified light chain polypeptide(s) of the antibody is conjugated to an agent or agents via covalent binding to no more than 2, 3, 4, 5, 6, 7, 8, 9 or 10 cysteines of the C-terminal extension.
- the extension includes an agent that is conjugated to two or more adjacent cysteine residues in the extension, either directly or via one or more linkers. For example, the extension may include an agent that is conjugated to two adjacent cysteine residues in the extension.
- the number of agents conjugated to the C-terminal extensions of modified light chain polypeptide(s) of the antibody can be characterized by the drug-to-antibody ratio (DAR).
- DAR drug-to-antibody ratio
- a distribution of a plurality of antibody conjugates may be characterized by measuring the average DAR of the antibody conjugates in the distribution, where the average DAR indicates the average number of agents conjugated to the C-terminal extensions of modified light chain polypeptide(s) of the antibodies in the distribution.
- average is meant the arithmetic mean. In certain cases, average DAR is assayed by
- an average DAR provides an indication of the average drug-to-antibody ratio of the antibody conjugates in a distribution provided by the DAR assay.
- the DAR of an antibody conjugate according to the present disclosure ranges from 0 to 20, such as from 1 to 17, or 1 to 15, or 1 to 12, or 1 to 10, or 1 to 9, or 1 to 8, or 1 to 7, or 1 to 6, or 1 to 5, or 1 to 4, or 1 to 3.
- a DAR of 0 indicates that the antibody is unconjugated; a DAR of 1 indicates that one agent (e.g., drug) is conjugated to a C- terminal extension of a modified light chain polypeptide of the antibody; a DAR of 2 indicates that two agents are conjugated to one or more C-terminal extensions of modified light chain polypeptide(s) of the antibody; a DAR of 3 indicates that three agents are conjugated to one or more C-terminal extensions of modified light chain polypeptide(s) of the antibody; a DAR of 4 indicates that four agents are conjugated to one or more C-terminal extensions of modified light chain polypeptide(s) of the antibody; etc.
- the DAR of an antibody conjugate according to the present disclosure is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.
- the average DAR of a distribution of antibody conjugates according to the present disclosure ranges from 0 to 20, such as from 1 to 17, or 1 to 15, or 1 to 12, or 1 to 10, or 1 to 9, or 1 to 8, or 1 to 7, or 1 to 6, or 1 to 5, or 1 to 4, or 1 to 3.
- an average DAR of 0 indicates that, on average, the antibodies in the distribution are unconjugated; an average DAR of 1 indicates that, on average, one agent (e.g., drug) is conjugated to each antibody in the distribution; an average DAR of 2 indicates that, on average, two agents are conjugated each antibody in the distribution; an average DAR of 3 indicates that, on average, three agents are conjugated to each antibody in the distribution; an average DAR of 4 indicates that, on average, four agents are conjugated to each antibody in the distribution; etc.
- an average DAR of a distribution of antibody conjugates according to the present disclosure is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.
- a sample containing antibody conjugates according to the present disclosure includes a mixture of distributions of unconjugated and conjugated (e.g., mono-conjugated, di-conjugated, tri-conjugated, etc.) antibodies.
- the average DAR of each distribution of antibody conjugates may be assayed (e.g., by HIC).
- An example antibody conjugate according to an embodiment of the present disclosure is schematically illustrated in FIG. 2.
- conjugate 200 includes an antibody having two light chain polypeptides that include light chain variable (V L ) and constant (C L ) domains, and C- terminal extensions having the sequence GGGSC extending from the C-terminal residue of each of the (C L ) domains.
- Conjugate 200 further includes agents 202 and 204 linked to the cysteine residue of the extension via a linker.
- the agent is conjugated to the cysteine residue via a linker.
- Linkers that find use in the conjugates of the present disclosure include maleimide or maleimide-based linkers; valine-citrulline linkers; hydrazone linkers; N-succinimidyl-4-(2-pyridyldithio)butyrate (SPDB) linkers; Succinimidyl-4-(N-maleimidomethyl)cyclohexane-l-carboxylate (SMCC) linkers; vinylsulfone-based linkers; linkers involving metal atom(s) coordinated to cysteine; linkers that include polyethylene glycol (PEG), such as, but not limited to tetraethylene glycol; linkers that include propanoic acid; linker that include caproleic acid; linkers that include Fleximer® polymers (Mersana Therapeutics, Cambridge, MA) or the linkers used to attach drugs to Fleximer polymers (see, e.g., USPN 8,
- the linker is a chemically-labile linker, such as an acid-cleavable linker that is stable at neutral pH (bloodstream pH 7.3-7.5) but undergoes hydrolysis upon
- Chemically-labile linkers include, but are not limited to, hydrazone-based linkers.
- the linker is an enzyme-labile linker, such as an enzyme-labile linker that is stable in the bloodstream but undergoes enzymatic cleavage upon internalization into a target cell, e.g., by a lysosomal protease (such as cathepsin or plasmin) in a lysosome of the target cell (e.g., a cancer cell).
- Enzyme-labile linkers include, but are not limited to, linkers that include peptidic bonds, e.g., valine-citrulline linkers, such as a maleimidocaproyl-valine-citruline-/?-aminobenzyl (MC-vc-PAB) linker, and the like.
- the linker is a non-cleavable linker, such as a linker that includes a non-cleavable thioether bond.
- Chemically-labile linkers, enzyme-labile, and non-cleavable linkers are known and described in detail, e.g., in Ducry & Stump (2010) Bioconjugate Chem. 21 :5-13.
- the linker is (or includes) a sulfhydryl-reactive chemical group capable of reacting with one or more reduced sulfhydryl group (or thiol, -SH) of the cysteine residue(s) of the C-terminal amino acid extension.
- Sulfhydryl-reactive chemical groups that find use in linking the agent to the cysteine of the C-terminal extension include, but are not limited to, maleimides, haloacetyls, pyridyl disulfides, aziridines, acryloyls, arylating agents, vinylsulfones, TNB-thiols, metals, and disulfide reducing agents.
- Such groups may conjugate to sulfhydryls by alkylation (e.g., by the formation of a thioether bond), disulfide exchange (formation of a disulfide bond), or the like.
- the agents which are the payload of the antibody conjugates of the present disclosure can be any suitable agent.
- the agent selected for use in the antibody conjugates of the present disclosure will vary depending on the application for which the conjugate is employed (e.g., killing, prevention of cell proliferation, hormone therapy, target imaging, and/or gene therapy, etc.).
- Agents of interest include, but are not limited to, therapeutic agents (e.g., drugs (e.g., cytotoxic agents)), detectable agents (e.g., in vivo imaging agents), and/or any other agent useful for a particular antibody-based application of interest.
- Non-limiting examples of such agents include toxins, fragments of toxins, lectins, alkylating agents, enzymes, antibiotics such as antibacterials, antifungals, antimycoplasmals, etc., antiviral agents, antimetabolites, antiproliferative or antineoplastic agents, DNA, radioopaque dyes, radioactive isotopes (e.g., I 123 , 1 131 as well as radioactive metal ions), metal ions, fluorogenic compounds, marker compounds, and compounds which alter cell membrane permeability.
- the agent is (or includes) a member of a specific binding pair, e.g., biotin (which forms a specific binding pair with avidin/streptavidin).
- the agent is a therapeutic agent.
- Therapeutic agents of interest include agents capable of affecting the function of a cell/tissue to which the conjugate binds via specific binding of the antibody portion of the conjugate to an antigen on the surface of the cell/tissue.
- the agent may boost the function of the cell/tissue to which the conjugate specifically binds.
- an agent that reduces the function of the cell/tissue may be employed.
- a conjugate of the present disclosure includes an agent that reduces the function of a target cell/tissue by inhibiting cell proliferation and/or killing the cell/tissue.
- agents may vary and include cytostatic agents and cytotoxic agents (e.g., an agent capable of killing a target cell tissue with or without being internalized into a target cell).
- the therapeutic agent is a cytotoxic agent selected from an enediyne, a lexitropsin, a duocarmycin, a taxane, a puromycin, a dolastatin, a maytansinoid, and a vinca alkaloid.
- the cytotoxic agent is paclitaxel, docetaxel, CC-1065, CPT-11 (SN-38), topotecan, doxorubicin, morpholino-doxorubicin, rhizoxin, cyanomorpholino- doxorubicin, dolastatin- 10, echinomycin, combretastatin, calicheamicin, maytansine, maytansine DM1, maytansine DM4, DM-1, an auristatin or other dolastatin derivatives, such as auristatin E or auristatin F, AEB (AEB-071), AEVB (5-benzoylvaleric acid-AE ester), AEFP (antibody- endostatin fusion protein), MMAE (monomethylauristatin E), MMAF (monomethylauristatin F), pyrrolobenzodiazepines (PBDs), eleutherobin, netropsin, or any
- the agent is a protein toxin selected from hemiasterlin and hemiasterlin analogs such as HTI-286 (e.g., see USPN 7,579,323; WO 2004/026293; and USPN 8,129,407, the full disclosures of which are incorporated herein by reference), abrin, brucine, cicutoxin, diphtheria toxin, batrachotoxin, botulism toxin, shiga toxin, endotoxin,
- HTI-286 e.g., see USPN 7,579,323; WO 2004/026293; and USPN 8,129,407, the full disclosures of which are incorporated herein by reference
- HTI-286 e.g., see USPN 7,579,323; WO 2004/026293; and USPN 8,129,407, the full disclosures of which are incorporated herein by reference
- abrin brucine
- cicutoxin diphtheria toxin
- Pseudomonas exotoxin Pseudomonas endotoxin, tetanus toxin, pertussis toxin, anthrax toxin, cholera toxin, falcarinol, fumonisin Bl, fumonisin B2, afla toxin, maurotoxin, agitoxin, charybdotoxin, margatoxin, slotoxin, scyllatoxin, hefutoxin, calciseptine, taicatoxin,
- calcicludine geldanamycin, gelonin, lotaustralin, ocratoxin A, patulin, ricin, strychnine, trichothecene, zearlenone, and tetradotoxin.
- Enzymatically active toxins and fragments thereof which may be employed include diphtheria A chain, non-binding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), Momordica charantia inhibitor, curcin, crotin, Sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin and the tricothecenes.
- diphtheria A chain non-binding active fragments of diphtheria toxin
- exotoxin A chain from Pseudomonas aeruginosa
- ricin A chain abrin A chain
- modeccin A chain alpha
- the agent is a labeling agent.
- labeling agent or “detectable label” is meant the agent detectably labels the antibody, such that the antibody may be detected in an application of interest (e.g., in vitro and/or in vivo research and/or clinical applications).
- Detectable labels of interest include radioisotopes, enzymes that generate a detectable product (e.g., horseradish peroxidase, alkaline phosphatase, etc.), fluorescent proteins, paramagnetic atoms, and the like.
- the antibody is conjugated to a specific binding partner of detectable label (e.g., conjugated to biotin such that detection may occur via a detectable label that includes avidin/stre tavidin).
- the agent is a labeling agent that finds use in in vivo imaging, such as near-infrared (NIR) optical imaging, single-photon emission computed tomography (SPECT)/CT imaging, positron emission tomography (PET), nuclear magnetic resonance (NMR) spectroscopy, or the like.
- NIR near-infrared
- SPECT single-photon emission computed tomography
- PET positron emission tomography
- NMR nuclear magnetic resonance
- Labeling agents that find use in such applications include, but are not limited to, fluorescent labels, radioisotopes, and the like.
- the labeling agent is a multi-modal in vivo imaging agent that permits in vivo imaging using two or more imaging approaches (e.g., see Thorp-Greenwood and Coogan (2011) Dalton Trans. 40:6129-6143).
- the labeling agent is an in vivo imaging agent that finds use in near- infrared (NIR) imaging applications, which agent is selected from a Kodak X-SIGHT dye, Pz 247, DyLight 750 and 800 Fluors, Cy 5.5 and 7 Fluors, Alexa Fluor 680 and 750 Dyes, IRDye 680 and 800CW Fluors.
- NIR near- infrared
- the labeling agent is an in vivo imaging agent that finds use in SPECT imaging applications, which agent is selected from 99m Tc,
- the labeling agent is an in vivo imaging agent that finds use in positron emission tomography (PET) imaging applications, which agent is selected from U C, 13 N, 15 0, 18 F, 64 Cu, 62 Cu, 124 I, 76 Br, 82 Rb and 68 Ga.
- PET positron emission tomography
- the present disclosure also provides methods of making antibody conjugates.
- the methods include conjugating an agent to an antibody that includes a light chain polypeptide including a C-terminal amino acid extension, which extension includes a cysteine residue, where the agent is conjugated to the cysteine residue (directly or indirectly (e.g., via a linker)) of the C- terminal amino acid extension.
- the method involves the preferential (or "biased") conjugation of agent to the cysteine residue of the C-terminal amino acid extension rather than a cysteine residue outside the C-terminal extension.
- the conjugation includes conjugating a linker to a sulfhydryl group of the cysteine residue, e.g., using maleimide reaction chemistry, haloacetyl reaction chemistry, pyridyl disulfide reaction chemistry, or any other suitable reaction chemistry as described herein.
- the methods of making the conjugate may further include reducing the cysteine residue to a sulfhydryl group (i.e., thiol) prior to the conjugating step, e.g., using a suitable reducing agent and reaction conditions as described above.
- Suitable reducing agents include, but are not limited to, DTP A, cysteamine, TCEP (tris(2-carboxyethyl)phosphine hydrochloride), combinations thereof, and the like.
- methods of making the conjugate include contacting the antibody that includes a light chain polypeptide including a C-terminal amino acid extension with a reducing agent.
- methods of making the conjugate include contacting the antibody that includes a light chain polypeptide including a C-terminal amino acid extension with a first reducing agent, followed by contacting the antibody that includes a light chain polypeptide including a C-terminal amino acid extension with a second reducing agent.
- an alternative embodiment of the present disclosure does not require a reduction step as the cysteine within the light chain extension is already in a reduced state as a synthesis product.
- the reduced antibody may be contacted with a suitable oxidizing agent.
- suitable oxidizing agents include, but are not limited to, dehydroascorbic acid (DHAA), and the like.
- the agent conjugated to the antibody may be any useful agent.
- the agent is a therapeutic agent or a labeling agent, which agents are described elsewhere herein.
- the agent is linked to the cysteine of the C-terminal extension using maleimide reaction chemistry.
- the maleimide group may react specifically with sulfhydryl groups when the pH of the reaction mixture is between pH 6.5 and 7.5, resulting in the formation of a stable thioether linkage.
- a maleimidyl-modified agent e.g., drug
- a reduced cysteine e.g., sulfhydryl or thiol group
- primary amines may compete with thiols for reaction with maleimides, and also increase the rate of hydrolysis of the maleimide group to a non-reactive maleamic acid.
- Maleimides do not react with tyrosines, histidines or methionines.
- Bioconjugation approaches that employ maleimide-based linkers are described in, e.g., in Hermanson, G.T., Bioconjugate Techniques, 2nd ed. San Diego, CA Academic Press 2008; Aslam & Dent, Bioconjugation: Protein Coupling Techniques for the Biomedical Sciences, London Macmillan Reference Ltd 1998; Kalia & Raines, Advances in Bioconjugation, Curr.
- the agent is linked to the cysteine of the C-terminal extension using haloacetyl reaction chemistry.
- a haloacetyl linker that includes an iodoacetyl or a bromoacetyl group is employed.
- haloacetyls react with sulfhydryl groups at physiologic pH. The reaction of the iodoacetyl group proceeds by nucleophilic substitution of iodine with a sulfur atom from a sulfhydryl group, resulting in a stable thioether linkage.
- the agent is linked to the cysteine of the C-terminal extension using pyridyl disulfide reaction chemistry.
- pyridyl disulfides react with sulfhydryl groups over a broad pH range (with pH 4 to 5 being optimal) to form disulfide bonds.
- a disulfide exchange occurs between the sulfhydryl group of the antibody and a 2-pyridyldithiol group of a 2-pyridyldithiol-modified agent.
- the cysteine may be contacted with a suitable reducing agent under conditions sufficient to produce a reduced sulfhydryl group.
- the reducing agent is selected from cysteamine hydrochloride, 2- mercaptoethanol, dithiothreitol (DTT), 2-mercaptoethylamine, tris(2-carboxyl)phosphine (TCEP), cysteine HC1, N-ethylmaleimide, Nacystelyn, dornase alfa, thymosin ⁇ 4, guaifenesin TCEP HC1, and any combination thereof.
- cysteamine hydrochloride 2- mercaptoethanol, dithiothreitol (DTT), 2-mercaptoethylamine, tris(2-carboxyl)phosphine (TCEP), cysteine HC1, N-ethylmaleimide, Nacystelyn, dornase alfa, thymosin ⁇ 4, guaifenesin TCEP HC1, and any combination thereof.
- Reaction conditions for such reducing agents are known in the art and may be optimized, e.g., to promote selectivity or "bias" the reduction of the cysteine(s) present in the C-terminal extension as opposed to the cysteine residues present in the parental antibody (e.g., the cysteine residues that participate in disulfide bonding between C L and C H I of the light and heavy chains, and/or between the hinge regions of the heavy chains).
- An alternative embodiment of the invention does not require a reduction step as the cysteine within the light chain extension is already in a reduced state as a synthesis product.
- Preferential reduction of the cysteine(s) of the C-terminal amino acid extension over one or more cysteine residues outside the C-terminal amino acid extension may be achieved by selection of suitable reduction conditions.
- suitable reduction conditions include suitable selection of one or more of the following: a mild reducing agent and/or a reducing agent having a steric bulk that confers upon the reducing agent a preference for reducing a cysteine of the C-terminal amino acid extension; concentrations of the reducing agent and substrate; the temperature at which the reduction reaction is carried out, the pH of the reduction reaction mixture; the buffer used in the reduction reaction; and/or conditions under which the cells expressing the extended C-terminal light chain polypeptides are cultured (e.g., to obtain free thiol on the C-terminal extension and/or to generate readily reduced intermolecular disulfides).
- compositions may include any of the antibodies, conjugates, nucleic acids, vectors, and/or host cells described above. Aspects of the present disclosure include pharmaceutical
- the pharmaceutical compositions include any of the antibodies described elsewhere herein (e.g., an antibody that includes a light chain polypeptide including a C-terminal amino acid extension that includes a cysteine residue as described above) or any of the conjugates described elsewhere herein (e.g., a conjugate that includes an antibody component having a light chain polypeptide including a C-terminal amino acid extension that includes a cysteine residue as described above), and a pharmaceutically acceptable excipient.
- the antibody or conjugate present in the pharmaceutical compositions may include any of the features described above with respect to the antibodies of the present disclosure or the conjugates of the present disclosure, in any combination.
- the C-terminal extension of the antibody, or antibody portion of the conjugate may include any of the C-terminal extension features described above with respect to the length of the extension, the amino acid makeup of the extension, the number of spacers in the extension and amino acid sequences thereof, extension configurations based on combinations of one or more spacers and one or more cysteine residues, and any other aspects of the C-terminal extensions described above and elsewhere herein.
- compositions generally include a therapeutically effective amount of an antibody or conjugate of the present disclosure.
- An effective amount may be administered in one or more administrations.
- the antibodies or conjugates of the present disclosure may be administered to the patient using any convenient means capable of resulting in the desired therapeutic effect or diagnostic effect.
- the antibody or conjugate can be incorporated into a variety of formulations for therapeutic administration. More particularly, the antibody can be formulated into
- compositions by combination with appropriate, pharmaceutically acceptable carriers or diluents, and may be formulated into preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, injections, inhalants and aerosols.
- Formulations of the antibodies or conjugates of the present disclosure suitable for administration to a patient are generally sterile and may further be free of detectable pyrogens or other contaminants contraindicated for administration to a patient according to a selected route of administration.
- the antibody in pharmaceutical dosage forms, can be administered in the form of their pharmaceutically acceptable salts, or they may also be used alone or in appropriate association, as well as in combination, with other pharmaceutically active compounds.
- the following methods and excipients are merely examples and are in no way limiting.
- the antibodies or conjugates can be used alone or in combination with appropriate additives to make tablets, powders, granules or capsules, for example, with conventional additives, such as lactose, mannitol, corn starch or potato starch; with binders, such as crystalline cellulose, cellulose derivatives, acacia, corn starch or gelatins; with disintegrators, such as corn starch, potato starch or sodium carboxymethylcellulose; with lubricants, such as talc or magnesium stearate; and if desired, with diluents, buffering agents, moistening agents, preservatives and flavoring agents.
- conventional additives such as lactose, mannitol, corn starch or potato starch
- binders such as crystalline cellulose, cellulose derivatives, acacia, corn starch or gelatins
- disintegrators such as corn starch, potato starch or sodium carboxymethylcellulose
- lubricants such as talc or magnesium stearate
- the antibodies or conjugates can be formulated into preparations for injection by dissolving, suspending or emulsifying them in an aqueous or non-aqueous solvent, such as vegetable or other similar oils, synthetic aliphatic acid glycerides, esters of higher aliphatic acids or propylene glycol; and if desired, with conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifying agents, stabilizers and preservatives.
- compositions of the present disclosure may be prepared by mixing the antibody or conjugate having the desired degree of purity with optional physiologically acceptable carriers, excipients, stabilizers, surfactants, buffers and/or tonicity agents.
- Acceptable carriers, excipients and/or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid, glutathione, cysteine, methionine and citric acid; preservatives (such as ethanol, benzyl alcohol, phenol, m-cresol, p-chlor-m-cresol, methyl or propyl parabens, benzalkonium chloride, or combinations thereof); amino acids such as arginine, glycine, ornithine, lysine, histidine, glutamic acid, aspartic acid, isoleucine, leucine, alanine,
- polypeptides such as polypeptides
- proteins such as gelatin or serum albumin
- chelating agents such as EDTA
- sugars such as trehalose, sucrose, lactose, glucose, mannose, maltose, galactose, fructose, sorbose, raffinose, glucosamine, N-methylglucosamine, galactosamine, and neuraminic acid
- non-ionic surfactants such as Tween, Brij Pluronics, Triton-X, or polyethylene glycol (PEG).
- the pharmaceutical composition may be in a liquid form, a lyophilized form or a liquid form reconstituted from a lyophilized form, wherein the lyophilized preparation is to be reconstituted with a sterile solution prior to administration.
- the standard procedure for reconstituting a lyophilized composition is to add back a volume of pure water (typically equivalent to the volume removed during lyophilization); however solutions comprising antibacterial agents may be used for the production of pharmaceutical compositions for parenteral administration.
- Example antibody or conjugate concentrations in a pharmaceutical composition according the present disclosure may range from about 1 mg/rnL to about 200 mg/ml or from about 50 mg/mL to about 200 mg/mL, or from about 150 mg/mL to about 200 mg/rnL.
- An aqueous formulation of the antibody or conjugate may be prepared in a pH-buffered solution, e.g., at pH ranging from about 4.0 to about 7.0, or from about 5.0 to about 6.0, or alternatively about 5.5.
- buffers that are suitable for a pH within this range include phosphate-, histidine-, citrate-, succinate-, acetate-buffers and other organic acid buffers.
- the buffer concentration can be from about 1 mM to about 100 mM, or from about 5 mM to about 50 mM, depending, e.g., on the buffer and the desired tonicity of the formulation.
- a tonicity agent may be included in the antibody or conjugate formulation to modulate the tonicity of the formulation.
- Example tonicity agents include sodium chloride, potassium chloride, glycerin and any component from the group of amino acids, sugars as well as combinations thereof.
- the aqueous formulation is isotonic, although hypertonic or hypotonic solutions may be suitable.
- the term "isotonic" denotes a solution having the same tonicity as some other solution with which it is compared, such as physiological salt solution or serum.
- Tonicity agents may be used in an amount of about 5 mM to about 350 mM, e.g., in an amount of 100 mM to 350 mM.
- a surfactant may also be added to the antibody or conjugate formulation to reduce aggregation of the formulated antibody or conjugate and/or minimize the formation of particulates in the formulation and/or reduce adsorption.
- Example surfactants include
- polyoxyethylensorbitan fatty acid esters Teween
- polyoxyethylene alkyl ethers Brij
- alkylphenylpolyoxyethylene ethers Triton-X
- polyoxyethylene-polyoxypropylene copolymer Polyoxyethylene-polyoxypropylene copolymer
- SDS sodium dodecyl sulfate
- polyoxyethylenesorbitan-fatty acid esters are polysorbate 20, (sold under the trademark Tween 20TM) and polysorbate 80 (sold under the trademark Tween 80TM).
- suitable polyethylene-polypropylene copolymers are those sold under the names Pluronic® F68 or Poloxamer 188TM.
- suitable Polyoxyethylene alkyl ethers are those sold under the trademark BrijTM.
- concentrations of surfactant may range from about 0.001% to about 1% w/v.
- a lyoprotectant may also be added in order to protect the active ingredient (e.g. the antibody or conjugate) against destabilizing conditions during the lyophilization process.
- active ingredient e.g. the antibody or conjugate
- known lyoprotectants include sugars (including glucose and sucrose); polyols
- Lyoprotectants can be included in an amount of about 10 mM to 500 nM.
- the formulation includes an antibody or conjugate of the present disclosure, and one or more of the above-identified agents (e.g., a surfactant, a buffer, a stabilizer, a tonicity agent) and is essentially free of one or more preservatives, such as ethanol, benzyl alcohol, phenol, m-cresol, p-chlor-m-cresol, methyl or propyl parabens, benzalkonium chloride, and combinations thereof.
- a preservative is included in the formulation, e.g., at concentrations ranging from about 0.001 to about 2% (w/v).
- the formulation can be a liquid (e.g., an aqueous solution or emulsion) or lyophilized formulation thereof, suitable for parenteral administration, and can comprise: about 1 mg/mL to about 200 mg/mL of a subject antibody or conjugate; about 0.001 % to about 1 % of at least one surfactant; about 1 mM to about 100 mM of a buffer; optionally about 10 mM to about about 4.0 to about 7.0.
- a liquid e.g., an aqueous solution or emulsion
- lyophilized formulation thereof suitable for parenteral administration, and can comprise: about 1 mg/mL to about 200 mg/mL of a subject antibody or conjugate; about 0.001 % to about 1 % of at least one surfactant; about 1 mM to about 100 mM of a buffer; optionally about 10 mM to about about 4.0 to about 7.0.
- An antibody or conjugate of the present disclosure can be utilized in an aerosol formulation to be administered via inhalation.
- the antibody can be formulated into pressurized acceptable propellants such as dichlorodifluoromethane, propane, nitrogen and the like.
- Unit dosage forms for oral administration such as syrups, elixirs, and suspensions may be provided wherein each dosage unit, for example, teaspoonful, tablespoonful, or tablet, contains a predetermined amount of the composition containing one or more inhibitors.
- unit dosage forms for injection or intravenous administration may comprise the antibody or conjugate in a composition as a solution in sterile water, normal saline or another pharmaceutically acceptable carrier.
- unit dosage form refers to physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of compounds of the present invention calculated in an amount sufficient to produce the desired effect in association with a pharmaceutically acceptable diluent, carrier or vehicle.
- the specifications for the antibody or conjugate of interest may depend on the particular antibody employed and the effect to be achieved, and the pharmacodynamics associated with each antibody in the host.
- the pharmaceutical composition (optionally provided in unit dosage form) includes an antibody or conjugate of the present disclosure present at a concentration of from about 10 mg/rnL to about 1000 mg/rnL, e.g., from about 25 mg/mL to about 500 mg/mL, from about 50 mg/mL to about 250 mg/mL, from about 75 mg/mL to about 200 mg/mL, or from about 100 mg/mL to about 150 mg/mL (e.g., about 125 mg/mL).
- the antibody or conjugate is formulated in a controlled release formulation.
- Sustained-release preparations may be prepared using methods well known in the art. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody in which the matrices are in the form of shaped articles, e.g. films or microcapsules. Examples of sustained-release matrices include polyesters, copolymers of L-glutamic acid and ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, hydrogels, polylactides, degradable lactic acid-glycolic acid copolymers and poly-D-(-)-3- hydroxybutyric acid. Controlled release within the scope of this invention can be taken to mean any one of a number of extended release dosage forms. The following terms may be considered to be substantially equivalent to controlled release, for the purposes of the present invention:
- a suitable dosage can be determined by an attending physician or other qualified medical personnel, based on various clinical factors. As is well known in the medical arts, dosages for any one patient depend upon many factors, including the patient's size, body surface area, age, the particular antibody or conjugate to be administered, sex of the patient, time, and route of administration, general health, and other drugs being administered concurrently.
- An antibody or conjugate of the present disclosure may be administered in amounts between 1 ng/kg body weight and 25 mg/kg body weight per dose, e.g. between 0.1 mg/kg body weight to 10 mg/kg body weight, e.g. between 0.5 mg/kg body weight to 8 mg/kg body weight, e.g. between 1 mg/kg body weight to 6 mg/kg body weight, e.g.
- the regimen is a continuous infusion, it can also be in the range of 1 ⁇ g to 10 mg per kilogram of body weight per minute.
- dose levels can vary as a function of the specific antibody, the severity of the symptoms and the susceptibility of the subject to side effects.
- Preferred dosages for a given compound are readily determinable by those of skill in the art by a variety of means.
- routes of administration include intravenous, intra-arterial, intramuscular, intranasal, intra-tracheal, subcutaneous, intradermal, topical application, nasal, oral, and other enteral and parenteral routes of administration. Routes of administration may be combined, if desired, or adjusted depending upon the antibody or conjugate and/or the desired effect.
- the pharmaceutical composition can be administered in a single dose or in multiple doses. In some embodiments, the composition is administered intravenously. In some embodiments, the composition is administered orally. In some embodiments, the composition is administered via an inhalational route. In some embodiments, the composition is administered intranasally. In some embodiments, the composition is administered locally. In some embodiments, the composition is administered intra-cranially.
- the present disclosure provides methods of treating diseases or disorders.
- the methods may include administering to a patient in need thereof a therapeutically effective amount of any of the antibodies, conjugates, or pharmaceutical compositions described elsewhere herein.
- the antibody or conjugate may be administered alone (e.g., in monotherapy) or in combination (e.g., in combination therapy) with one or more additional therapeutic agents.
- an effective amount of the antibody or conjugate is an amount that, when administered alone (e.g., in monotherapy) or in combination (e.g., in combination therapy) with one or more additional therapeutic agents, in one or more doses, is effective to reduce the symptoms of a disease or disorder in an individual by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to the symptoms in the individual in the absence of treatment with the antibody or conjugate.
- an antibody or conjugate of the present disclosure inhibits growth, metastasis and/or invasiveness of a cancer cell(s) in a host when the antibody or conjugate is administered in an effective amount.
- cancer cell is meant a cell exhibiting a neoplastic cellular phenotype, which may be characterized by one or more of, for example, abnormal cell growth, abnormal cellular proliferation, loss of density dependent growth inhibition, anchorage- independent growth potential, ability to promote tumor growth and/or development in an immunocompromised non-human animal model, and/or any appropriate indicator of cellular transformation.
- Cancer cell may be used interchangeably herein with “tumor cell”, “malignant cell” or “cancerous cell”, and encompasses cancer cells of a solid tumor, a semi-solid tumor, a primary tumor, a metastatic tumor, and the like.
- the antibody or antibody component of the conjugate specifically binds to an antigen on the surface of a cancer cell.
- the terms “antigen” and “epitope” are well understood in the art and refer to the portion of a macromolecule (e.g., a polypeptide) which is specifically recognized by a component of the immune system, e.g., an antibody or a T-cell antigen receptor.
- the term "antigen” encompasses antigenic epitopes, e.g., fragments of an antigen which are antigenic epitopes.
- Haptens are also examples of antigens.
- Epitopes can be recognized by antibodies in solution, e.g. free from other molecules.
- Epitopes can be recognized by T-cell antigen receptor when the epitope is associated with a class I or class II major histocompatibility complex molecule.
- Antigens of interest in the context of cancer treatment include tumor-specific antigens, e.g., antigens present on the surface of malignant cells and not present on non-malignant cells.
- the antigen bound by the antibody is a tumor-associated antigen.
- tumor-associated antigen is meant an antigen expressed on malignant cells with limited expression on cells of normal tissues, antigens that are expressed at much higher density on malignant versus normal cells, or antigens that are developmentally expressed.
- any tumor-associated antigen or tumor-specific antigen may be targeted by an antibody or conjugate of the present disclosure.
- the antigen specifically bound by the antibody or antibody component of a conjugate of the present disclosure may include, but is not limited to, HER2, CD19, CD22, CD30, CD33, CD56, CD66/CEACAM5, CD70, CD74, CD79b, CD138, Nectin-4, Mesothelin, Transmembrane glycoprotein NMB (GPNMB), Prostate-Specific Membrane Antigen (PSMA), SLC44A4, CA6, CA-IX, or any other tumor-associated or tumor-specific antigens of interest.
- GPNMB Transmembrane glycoprotein NMB
- PSMA Prostate-Specific Membrane Antigen
- SLC44A4 CA6, CA-IX, or any other tumor-associated or tumor-specific antigens of interest.
- telomere binding or telomere binding in the context of a characteristic of an antibody refers to the ability of an antibody to preferentially bind to a particular antigen that is present in a homogeneous mixture of different antigens.
- a specific binding interaction will discriminate between desirable and undesirable antigens (or “target” and “non-target” antigens) in a sample or organism (e.g., a human), in some embodiments more than about 10 to 100-fold or more (e.g., more than about 1000- or 10,000-fold).
- the affinity between an antibody and antigen when they are specifically bound in an antibody-antigen complex is characterized by a KD (dissociation constant) of less than 10 "6 M, less than 10 "7 M, less than 10 "8 M, less than 10 "9 M, less than 10 "10 M, less than 10 "11 M, or less than about 10 "12 M or less.
- KD dissociation constant
- Cancers which may be treated using the methods of the present disclosure include, but are not limited to, solid tumors, breast cancer, prostate cancer, pancreatic cancer, colorectal carcinoma, renal cell carcinoma, Hodgkin's lymphoma, Non-Hodgkin's lymphoma, anaplastic large cell lymphoma, acute myelogenous leukemia, multiple myeloma, and any other type of cancer which may be treated using an antibody-based or antibody-conjugate-based therapy.
- kits may include any of the antibodies, conjugates, or pharmaceutical compositions of the present disclosure having any of the features as described elsewhere herein. Alternatively, or
- kits may include any reagents useful for producing an antibody of the present disclosure or light chain polypeptide thereof, or any of the conjugates of the present disclosure.
- the kit may include a nucleic acid that encodes an antibody light chain polypeptide that include a cysteine-containing C-terminal amino acid extension.
- kits may include, e.g., competent cells or cells already harboring nucleic acids encoding one or more antibody light and/or heavy chain polypeptides, a reducing agent for reducing the sulfhydryl group of a cysteine residue in the C-terminal light chain polypeptide extension, a linker for conjugating an agent to a reduced sulfhydryl of a cysteine residue, an agent (which may be attached to a linker or separate from a linker), reagents, buffers, purification columns, etc. that find use in producing an antibody or conjugate of the present disclosure, or any combinations thereof.
- the kits find use, e.g., in enabling one to practice the methods of the present disclosure, such as the methods of treating a disease or disorder, methods of making antibody light chain polypeptides, and/or methods of making antibody conjugates.
- kits for practicing the methods may include one or more pharmaceuticals
- kits may include a single pharmaceutical composition present as one or more unit dosages. In yet other embodiments, the kits may include two or more separate pharmaceutical compositions.
- kits may be present in separate containers, or multiple components may be present in a single container. In certain embodiments, it may be convenient to provide the components in a lyophilized form, so that they are ready to use and can be stored conveniently at room temperature.
- a kit of the present disclosure may further include instructions for using the components of the kit, e.g., to treat a disease or disorder using an antibody or conjugate of the present disclosure, or to make an antibody or conjugate of the present disclosure.
- the instructions are generally recorded on a suitable recording medium.
- the instructions may be printed on a substrate, such as paper or plastic, etc.
- the instructions may be present in the kits as a package insert, in the labeling of the container of the kit or components thereof (i.e., associated with the packaging or subpackaging) etc.
- the instructions are present as an electronic storage data file present on a suitable computer readable storage medium, e.g.
- the actual instructions are not present in the kit, but means for obtaining the instructions from a remote source, e.g. via the internet, are provided.
- An example of this embodiment is a kit that includes a web address where the instructions can be viewed and/or from which the instructions can be downloaded. As with the instructions, this means for obtaining the instructions is recorded on a suitable substrate.
- a restriction site was introduced into the 5' end of the constant region.
- the heavy chain constant region sequence was changed from GCC TCC to GCT AGC which introduced a Nhel restriction site while maintaining the original amino acid sequence.
- the light chain polypeptide constant region sequence was changed from CGA ACT to CGT ACG to generate a BsiWI restriction site while maintaining the original amino acid sequence. The changes were introduced by designing mismatch PCR primers purchased from Operon - Euro fins.
- the IgkC 5' primer was designed with a 5' Nhel overhang to facilitate cloning into the vector.
- the 3' primers were designed with a BamHI restriction site 3 ' of the stop codon.
- the fragments and the vectors were digested using the relevant restriction enzymes and separated on a 1% agarose gel.
- the digested fragments were extracted from the gel using Qiagen gel extraction kit and ligated into the vector using T4 DNA ligase (New England Biolabs). Competent DH5a E-coli (Invitrogen) were transformed and single-cell colonies were grown at 37°C over night on ampicillin selective LB plates.
- Competent DH5a E-coli Competent DH5a E-coli (Invitrogen) were transformed and single-cell colonies were grown at 37°C over night on ampicillin selective LB plates.
- To isolate the plasmids single cells colonies were inoculated into liquid LB-ampicillin media, grown overnight at 37°C, and plasmids were isolated using Qiagen® QIAprep® spin miniprep kit. The clones were screened by miniprep DNA digests, and verified by sequence analysis using Geneious sequence alignment, assembly and analysis software from Biomatters.
- herceptin V-genes were synthesized as gBlocks® gene fragments from IDT® (Integrated DNA Technologies). The sequences synthesized are provided in Table 2.
- the fragments were cloned into the mentioned constant region pTT5 vectors using EcoRI and Nhel digests for the heavy chain and EcoRI and BsiWI digests for the light chain
- HEK293 cells were co-transfected with the Herceptin heavy chain and the Herceptin light chain-cys constructs using 293 fectin (Invitrogen). The cells were grown in Freestyle 293 expression media (Gibco) supplemented with 0.1% Pluronic F68 (Gibco) solution for 5 days. 24 hours post transfection the cells were supplemented with 0,5% tryptone.
- the eluates were assayed using a bicinchoninic acid assay (Pierce, #23225) using Herceptin as a standard to establish protein concentration, and with Ellman's reagent using cysteine as a standard to establish the absence of free thiol groups.
- Herceptin or Herceptin VLCysX (5-30 ⁇ in 100 mM phosphate, 50 mM NaCl, 2 mM DTPA, pH 6.1) was reduced by addition of cysteamine hydrochloride (from 5 to 10 mM ) from a 1.0 M stock in the same buffer and incubation for 40-180 minutes at either room temperature or 37 °C. After cooling to room temperature cysteamine was removed from the reaction mixture by passage over a ZebaTM spin column (40 KDa MWCO) preconditioned with 100 mM phosphate, 50 mM NaCl, 2 mM DTPA, pH 6.1.
- the eluate was assayed with Ellman's reagent employing a standard curve generated by assay of cysteine serial dilutions. This assay also provided some measure of the average thiol content per protein.
- Toxin 3 as used herein is MT-vc-toxin, where the toxin is (S,E)-N-(4- aminophenylsulfonyl)-2,5-dimethyl-4-((S)-N,3,3-trimethyl-2-((S)-3-methyl-2-(methylamino)-3- phenylbutanamido)butanamido)hex-2-enamide .
- Toxin 5" as used herein is MT-vc-P ABC-toxin, where the toxin is (S,E)-N-(4- (aminomethyl)benzylsulfonyl)-2,5-dimethyl-4-((S)-N,3,3-trimethyl-2-((S)-3-methyl-2- (methylamino)-3-phenylbutanamido)butanamido)hex-2-enamide.
- Toxin 6 as used herein is MT-vc-toxin, where the toxin is:
- HCC1954 cells 100 ⁇ were added to opaque-walled clear- bottomed 96-well tissue culture-treated microtiter plates using complete growth medium at a density of 2500 cells/100 ⁇ of medium.
- the HCC1954 cells were incubated for one night at 37° C/5% C02 to allow the cells to attach to the microtiter plate surface.
- Antibody drug conjugates were diluted in complete growth medium at five-times the final maximum
- Herceptin-Igk-Ig extension4 Herceptin VL and hlgk constant region (Km3
- Herceptin-Igk-Ig extension5 Herceptin VL and hlgk constant region (Km3 WKVDNALQSGNSQESVTEQ allotype) + a ERKCCVECPPC tail
- Herceptin-Igk-Ig extension6 Herceptin VL and hlgk constant region (Km3 WKVDNALQSGNSQESVTEQ allotype) + a ERKC tail (sequence from
- Herceptin-Igk-Ig extension Herceptin VL and hlgk constant region (Km3 WKVDNALQSGNSQESVTEQ allotype) + a DVITMDPKDNC tail
- SEQ ID NO: 110 (sequence from human TCRg hinge) DSKDSTYSLSSTLTLSKADY
- Herceptin VL and hlgk constant region Km3
- Herceptin-Igk-Ig extension9 Herceptin VL and hlgk constant region (Km3 WKVDNALQSGNSQESVTEQ allotype) + a ESSC tail (sequence from
- Herceptin-Igk-Ig extension 10 Herceptin VL and hlgk constant region (Km3 WKVDNALQSGNSQESVTEQ allotype) + a ESSCDVKLV tail (sequence
- Herceptin-Igk-Ig extension 11 Herceptin VL and hlgk constant region (Km3 WKVDNALQSGNSQESVTEQ allotype) + a DHVKPKETENTKQPSKSC DSKDSTYSLSSTLTLSKADY SEQ ID NO: 114 tail (sequence from human TCRd hinge) EKHKVYACEVTHQGLS SPVT
- Herceptin-Igk-Ig extension 12 Herceptin VL and hlgk constant region (Km3 WKVDNALQSGNSQESVTEQ allotype) + a DVITMDPKDNCSKDAN tail DSKDSTYSLSSTLTLSKADY SEQ ID NO: 115 (sequence from human TCRg hinge) EKHKVYACEVTHQGLS SPVT
- Herceptin-Igk-Ig extension2 Herceptin VL and hlgk constant region (Km3 GTCACAGAGCAGGACAGCA allotype) + a EPKSCDKTHTCPPC tail
- Herceptin-Igk-Ig extension3 Herceptin VL and hlgk constant region (Km3 GGGTAACTCCCAGGAGAGT allotype) + a EPKSC tail (sequence from GTCACAGAGCAGGACAGCA SEQ ID NO: 118 human IgGl hinge) AGGACAGCACCTACAGCCT
- Herceptin-Igk-Ig extension4 Herceptin VL and hlgk constant region (Km3 GGCCAGGGCACCAAGGTGG allotype) + a ESKYGPPC tail (sequence AGATCAAGCGTACGGTGGC Name Description Sequence (NT)
- Herceptin-Igk-Ig extension5 Herceptin VL and hlgk constant region (Km3
- Herceptin-Igk-Ig extension6 Herceptin VL and hlgk constant region (Km3 AGATCAAGCGTACGGTGGC allotype) + a ERKC tail (sequence from
- Herceptin-Igk-Ig extension Herceptin VL and hlgk constant region (Km3
- Herceptin VL and hlgk constant region Km3
- Herceptin-Igk-Ig extension9 Herceptin VL and hlgk constant region (Km3 GGGTAACTCCCAGGAGAGT allotype) + a ESSC tail (sequence from GTCACAGAGCAGGACAGCA SEQ ID NO: 124 human TCRa hinge) AGGACAGCACCTACAGCCT
- Herceptin-Igk-Ig extension 10 Herceptin VL and hlgk constant region (Km3 GTTGAAATCTGGAACTGCC allotype) + a ESSCDVKLV tail (sequence
- Herceptin-Igk-Ig extension 11 Herceptin VL and hlgk constant region (Km3 GTCACAGAGCAGGACAGCA allotype) + a DHVKPKETENTKQPSKSC AGGACAGCACCTACAGCCT SEQ ID NO: 126 tail (sequence from human TCRd hinge) CAGCAGCACCCTGACGCTG
- Herceptin-Igk-Ig extension 12 Herceptin VL and hlgk constant region (Km3 TCTGTTGTGTGCCTGCTGAA allotype) + a DVITMDPKDNCSKDAN tail
- SEQ ID NO: 127 sequence from human TCRg hinge
- HEK293 cells were co-transfected with the Herceptin heavy chain and the Herceptin light chain-cys constructs using 293 fectin (Invitrogen). The cells were grown in Freestyle 293 expression media (Gibco) supplemented with 0.1% Pluronic F68 (Gibco) solution for 5 days. 24 hours post transfection the cells were supplemented with 0.5% tryptone.
- Supematants were collected and the secreted antibodies were purified from supernatant using the AKTAxpress machine and HiTrap Mab Select SuRe columns (cat# 11-0034-93) followed by buffer exchange into PBS PH 7.4 using an Amicon® Ultra 15 filter with a 30 kDa MW cutoff (Millipore).
- Herceptin VLSpacerX (30 ⁇ in PBS) was reduced by addition of 1.25 mM
- DTPA diethylenetriaminepentaacetate
- maleimide toxin (toxin 1, toxin 3, or toxin 4) was added from a
- TCEP tris(2-carboxyethyl)phosphine hydrochloride
- DTP A a final antibody concentration of 2.5 mg/mL.
- the reduced light chain extension antibody was loaded onto a ZebaTM Spin Desalting Column, 40K MWCO, and eluted with PBS.
- the eluted reduced antibody was treated with 10 equivalents of 10 mM dehydroascorbic acid (DHAA) in PBS at room temperature, for 30 minutes.
- DHAA dehydroascorbic acid
- the reoxidized antibodies from Example 9 were combined with 3.5 molar equivalents relative to the antibody, mixed, and let stand for about an hour at room temperature to effect conjugation and form the light chain extension antibody-drug conjugate (ADC), including Tsp2- Toxin 3, Tsp3-Toxin 3, Tsp4-Toxin 3, Tsp5-Toxin 3, Tsp6-Toxin 3, Tsp9-Toxin 3, TsplO-Toxin 3, TsplO-Toxin 4, TsplO-Toxin 1, Tspl l-Toxin 3, TVLCysl-Toxin 3 (DAR 1.06), BsplO-Toxin 3, BsplO-Toxin 4, BsplO-Toxin 1, BsplO-Toxin 6, and BsplO-MC-vc-PABC-MMAE.
- ADC light chain extension antibody-drug conjugate
- HER2 expressing MDA-MB-231 cells were trypsinized and counted, and 50,000 cells per sample were incubated with unconjugated MAbs or with conjugated ADCs for 24 hours at 4°C in 50 ⁇ 1 total volume.
- Antibodies were applied at 20000, 4000, 800, 160, 32, 6.4, 1.28 and 0.256 ng/ml in Leibovitz's LI 5 media supplemented with 10% Fetal bovine serum. Following incubation the cells were washed twice in ice cold PBS+1% FBS and incubated with Alexa 647 labelled Goat anti-Human IgGFc (2ug/mL) secondary antibodies + 2.5ug/mL 7- Aminoactinomycin D.
- TSP2 is an antibody having the light chain polypeptide of SEQ ID NO: 105.
- TSP3 is an antibody having the light chain polypeptide of SEQ ID NO: 106.
- TSP4 is an antibody having the light chain polypeptide of SEQ ID NO: 107.
- TP5 is an antibody having the light chain polypeptide of SEQ ID NO: 108.
- TP6 is an antibody having the light chain polypeptide of SEQ ID NO: 109.
- TP9 is an antibody having the light chain polypeptide of SEQ ID NO: 112.
- TP10 is an antibody having the light chain polypeptide of SEQ ID NO: 113.
- TP11 is an antibody having the light chain polypeptide of SEQ ID NO: 114.
- VLcysl is trastuzumab having the C-terminal light chain extension GGGSC (SEQ ID NO:60).
- TSP10 Trastsuzumab with extension 10
- T Trastuzumab
- T Trastuzumab
- T Trastuzumab
- T Trastuzumab
- T Trastuzumab
- T Trastuzumab
- T Trastuzumab
- T Trastuzumab
- T Trastuzumab
- T Trastuzumab with extension 10
- T Trastuzumab
- 5-maleimido-Alexa488 was conjugated to MAbs using the reduction/conjugation methods described above.
- the antibodies were analyzed by SDS PAGE using the samples at a dilution between 1 : 15 to 1 :40 from 100 ⁇ g/ml in PBS and loading 20 ⁇ in each lane.
- the gels were imaged using a Typhoon TrioTM imager (GE Healthcare Life Sciences) measuring the fluorescence of Alexa488. Results are shown in FIG. 7.
- mice 7-8 week old Female NOD/SCID gamma (NSG) mice (Jackson) was inoculated with 5* 106 NCI-N87 tumor cells (ATCC Cat # CRL-5822) mixed 1 : 1 with matrigel in a total volume of ⁇ . Tumors were measured every Monday, Wednesday, and Friday. Once tumors reached 150-200 mm 3 in size, animals were assigned to treatment groups as shown in Table 6 below to counterbalance the average tumor size across groups. "T” is an abbreviation for trastuzumab. Table 6 - In vivo study groups
- HIC analysis of antibody drug conjugates were performed on a HP 1100 Series HPLC with a DAD at 280nm using a TSKgel Butyl-NPR column (2.5uM, 4.6mm x 3.5cm). The method was run at 1 mL/min from a linear gradient of 95% to 5% mobile phase A over 12 minutes with re-equilibration back to 95% A for 3 minutes (A: 1.5M ammonium sulphate and 25mM sodium phosphate monobasic at pH 4.4; B: 25% IPA in 25mM sodium phosphate at pH 4.73).
- Chemstation software was used for data collection, analysis and peak area quantification.
- Non-denaturing SEC analysis of recombinant antibodies and antibody drug conjugates were carried out on a HP 1100 Series HPLC with DAD at 280nm using an Acquity UPLC BEH 200 SEC column (1.7uM, 4.6mm x 150cm). The analysis was performed using an isocratic elution over 20 minutes at 0.2mL/min with 25mM sodium phosphate and 150mM sodium chloride buffer at pH 6.8. Chemstation software was used for data collection, analysis and peak area quantification.
- TSplO, TSplO-Toxin 3, TSplO-Toxin 1, TSplO- Toxin 4, TSp6-Toxin 3, TSp4-Toxin 3 in PBS, pH 7, were prepared.
- Non-denaturing SEC-UV analysis (described previously) was performed at an injection volume of 10 ⁇ at time zero prior to incubation at 37 °C. Aliquots of each sample were analyzed by non-denaturing SEC-UV after 191 hours of incubation, and after 330 hours of incubation. Percent monomer peak area of each species was adjusted against their respective zero time point measurement.
- trastuzumab control 99.36% of the protein sample was in the monomeric state, while one different aggregate species was present at 0.63%.
- T-VLCysl 98.39%> of the protein sample was in the monomeric state, while other aggregate species were present at 1.16%, 0.266% and 0.175%.
- T-SP2 96.5% of the protein sample was in the monomeric state, while two different aggregate species were present at 0.3%>, 1.5% and 1.8%.
- T-SP3 98.5% of the protein sample was in the monomeric state, while two different aggregate species were present at 1.17% and 0.35%.
- T-SP4 98% of the protein sample was in the monomeric state, while other different aggregate species were present at 1.4%, 0.39% and 0.22%.
- T-SP5 94% of the protein sample was in the monomeric state, while two different aggregate species were present at 2.1% and 3.2%.
- T-SP6 89% of the protein sample was in the monomeric state, while other different aggregate species were present at 1.5% and 8.8%.
- T-SP7 9.95%) of the protein sample was in the monomeric state, while other different aggregate species were present at 3%, 0.51% and 0.50%.
- T-SP9 94.5% of the protein sample was in the monomeric state, while other different aggregate species were present at 0.6%, 3.8% and 1%.
- T-SP10 97.3% of the protein sample was in the monomeric state, while other different aggregate species were present at 2.2%, 0.27% and 0.22%.
- Results for an antibody (T-VLcysl) having the extension of SEQ ID NO: 60 are shown in FIG. 9, Panels A-C. Results for an antibody (T-VLcys2) having the extension of SEQ ID NO:63 are shown in FIG. 10, Panels A-C. Results for an antibody (T-VLcys4) having the extension of SEQ ID NO:64 are shown in FIG. 11, Panels A and B. Results for an antibody having the light chain of SEQ ID NO: 105 are shown in FIG. 12, Panels A and B. Results for an antibody having the light chain of SEQ ID NO: 106 are shown in FIG. 13, Panels A-C. Results for an antibody having the light chain of SEQ ID NO: 107 are shown in FIG. 14, Panels A-C. Results for an antibody having the light chain of SEQ ID NO: 108 are shown in FIG. 15, Panels A and B.
- Results for an antibody having the light chain of SEQ ID NO: 109 are shown in FIG. 16, Panels A-C.
- Results for an antibody having the light chain of SEQ ID NO: 110 are shown in FIG. 17, Panels A-C.
- Results for an antibody having the light chain of SEQ ID NO: 113 are shown in FIG. 18, Panels A-C.
- Results for an antibody having the light chain of SEQ ID NO: 114 are shown in FIG. 19, Panels A-C.
- In vitro cell proliferation assays were performed using a procedure similar to that described in Example 5 above, by treating HER2 expressing HCC1954 cells, HER2 expressing N87 cells, and HER2 antigen negative Jurkat cells with various trastuzumab ("T")-based ADCs and controls.
- "Free Toxin 1" is Toxin 3 as defined above in its free form (i.e., not conjugated to an antibody). Results are shown in FIGs. 42-53 and summarized below in Tables 7 and 8. Table 7 - In vitro cell proliferation assay results (HCC1954 and Jurkat cells)
- Trastuzumab light chain extension variants were purified on immobilized protein A and subjected to non-reducing denaturing or reducing (+DTT) denaturing polyacrylamide gel electrophoresis (PAGE). Results are shown in FIGs. 59-62.
- ADC stability was assessed using a thermal stability assay. Individual aliquots of Img/mL solutions of TSplO, TSplO-Toxin 3, TSplO-Toxin 1 , TSplO-Toxin 4, TSp6-Toxin 3, TSp9-Toxin 3 in PBS, pH 7, were prepared. Non-denaturing SEC-UV analysis (described previously) was performed at an injection volume of 10 ⁇ at time zero prior to incubation at 37 °C. Aliquots of each sample were analyzed by non-denaturing SEC-UV after 191 hours of incubation, and after 330 hours of incubation. Percent monomer peak area of each species was adjusted against their respective zero time point measurement.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmacology & Pharmacy (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Biomedical Technology (AREA)
- Oncology (AREA)
- Cell Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
Claims
Priority Applications (12)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201480075958.5A CN106459981A (en) | 2013-12-23 | 2014-12-23 | Antibodies comprising C-terminal light chain polypeptide extensions and conjugates and methods of use thereof |
SG11201605093VA SG11201605093VA (en) | 2013-12-23 | 2014-12-23 | Antibodies comprising c-terminal light chain polypeptide extensions and conjugates and methods of use thereof |
AU2014373593A AU2014373593B2 (en) | 2013-12-23 | 2014-12-23 | Antibodies comprising C-terminal light chain polypeptide extensions and conjugates and methods of use thereof |
RU2016129724A RU2016129724A (en) | 2013-12-23 | 2014-12-23 | ANTIBODIES CONTAINING C-END LENGTH OF LIGHT CHAIN POLYPEPTIDE, THEIR CONJUGATES AND METHODS OF APPLICATION |
JP2016543004A JP6585600B2 (en) | 2013-12-23 | 2014-12-23 | Antibodies containing C-terminal light chain polypeptide extensions, and conjugates and methods of use thereof |
MX2016008355A MX2016008355A (en) | 2013-12-23 | 2014-12-23 | Antibodies comprising c-terminal light chain polypeptide extensions and conjugates and methods of use thereof. |
EP14874786.8A EP3087185A4 (en) | 2013-12-23 | 2014-12-23 | Antibodies comprising c-terminal light chain polypeptide extensions and conjugates and methods of use thereof |
BR112016014913A BR112016014913A8 (en) | 2013-12-23 | 2014-12-23 | antibody, or an antigen-binding fragment thereof, conjugate, nucleic acid, vector, host cell, pharmaceutical composition, method of preparing an antibody light chain or antigen-binding fragment, and method of preparing a conjugate |
CA2934818A CA2934818C (en) | 2013-12-23 | 2014-12-23 | Antibodies comprising c-terminal light chain polypeptide extensions and conjugates and methods of use thereof |
US15/107,884 US11208497B2 (en) | 2013-12-23 | 2014-12-23 | Antibodies comprising C-terminal light chain polypeptide extensions and conjugates and methods of use thereof |
KR1020167019769A KR20160122127A (en) | 2013-12-23 | 2014-12-23 | Antibodies comprising c-terminal light chain polypeptide extensions and conjugates and methods of use thereof |
IL246371A IL246371A0 (en) | 2013-12-23 | 2016-06-21 | Antibodies comprising c-terminal light chain polypeptide extensions and conjugates and methods of use thereof |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201361920425P | 2013-12-23 | 2013-12-23 | |
US61/920,425 | 2013-12-23 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2015095972A1 true WO2015095972A1 (en) | 2015-07-02 |
Family
ID=53477259
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/CA2014/051263 WO2015095972A1 (en) | 2013-12-23 | 2014-12-23 | Antibodies comprising c-terminal light chain polypeptide extensions and conjugates and methods of use thereof |
Country Status (13)
Country | Link |
---|---|
US (1) | US11208497B2 (en) |
EP (1) | EP3087185A4 (en) |
JP (1) | JP6585600B2 (en) |
KR (1) | KR20160122127A (en) |
CN (1) | CN106459981A (en) |
AU (1) | AU2014373593B2 (en) |
BR (1) | BR112016014913A8 (en) |
CA (1) | CA2934818C (en) |
IL (1) | IL246371A0 (en) |
MX (1) | MX2016008355A (en) |
RU (1) | RU2016129724A (en) |
SG (1) | SG11201605093VA (en) |
WO (1) | WO2015095972A1 (en) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11834506B2 (en) | 2017-02-08 | 2023-12-05 | Dragonfly Therapeutics, Inc. | Multi-specific binding proteins that bind NKG2D, CD16, and a tumor-associated antigen for activation of natural killer cells and therapeutic uses thereof to treat cancer |
US11884733B2 (en) | 2018-02-08 | 2024-01-30 | Dragonfly Therapeutics, Inc. | Antibody variable domains targeting the NKG2D receptor |
US11884732B2 (en) | 2017-02-20 | 2024-01-30 | Dragonfly Therapeutics, Inc. | Proteins binding HER2, NKG2D and CD16 |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2015164723A1 (en) | 2014-04-25 | 2015-10-29 | The Trustees Of The University Of Pennsylvania | Methods and compositions for treating metastatic breast cancer and other cancers in the brain |
EP3526257A2 (en) * | 2016-10-14 | 2019-08-21 | Dana-Farber Cancer Institute, Inc. | Modular tetravalent bispecific antibody platform |
WO2019014650A1 (en) * | 2017-07-13 | 2019-01-17 | City Of Hope | Anti-cancer phosphorothioate-coupled peptide conjugates and methods of using the same |
US11541121B2 (en) | 2017-07-13 | 2023-01-03 | City Of Hope | Phosphorothioate-conjugated peptides and methods of using the same |
MX2020002626A (en) * | 2017-09-07 | 2020-10-07 | Dragonfly Therapeutics Inc | Proteins binding nkg2d, cd16 and a tumor-associated antigen. |
JP2021522785A (en) * | 2018-05-01 | 2021-09-02 | アンブルックス, インコーポレイテッドAmbrx, Inc. | Methods for optimizing antibody expression |
ES2943474T3 (en) | 2018-11-30 | 2023-06-13 | Bristol Myers Squibb Co | Antibody comprising a carboxy-terminal extension of a glutamine-containing light chain, conjugates thereof, and methods and uses |
CN115947830B (en) * | 2022-12-27 | 2023-09-12 | 优洛生物(上海)有限公司 | Engineered antibodies, methods of making and uses thereof |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2012059882A2 (en) * | 2010-11-05 | 2012-05-10 | Rinat Neuroscience Corporation | Engineered polypeptide conjugates and methods for making thereof using transglutaminase |
Family Cites Families (23)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
RU2170589C2 (en) | 1992-10-28 | 2001-07-20 | Генентек Инк. | Composition for inhibition of angiogenesis, monoclonal antibody, polypeptide, method of inhibition of tumor growth (variants) |
US6652863B1 (en) * | 1992-11-16 | 2003-11-25 | Centocor, Inc. | Method of reducing the immunogenicity of compounds |
US6165476A (en) * | 1997-07-10 | 2000-12-26 | Beth Israel Deaconess Medical Center | Fusion proteins with an immunoglobulin hinge region linker |
US20050159343A1 (en) | 1999-03-26 | 2005-07-21 | Board Of Regents, University Of Texas System | Inhibitors of glycosaminoglycans |
WO2002028408A2 (en) | 2000-10-02 | 2002-04-11 | Arizeke Pharmaceuticals, Inc. | Compositions and methods for the transport of biologically active agents across cellular barriers |
US7834258B2 (en) * | 2003-06-30 | 2010-11-16 | Mu-Hyeon Choe | Dimer of chimeric recombinant binding domain-functional group fusion formed via disulfide-bond-bridge and the processes for producing the same |
WO2007103288A2 (en) * | 2006-03-02 | 2007-09-13 | Seattle Genetics, Inc. | Engineered antibody drug conjugates |
US8409577B2 (en) * | 2006-06-12 | 2013-04-02 | Emergent Product Development Seattle, Llc | Single chain multivalent binding proteins with effector function |
PE20140625A1 (en) * | 2007-07-16 | 2014-05-29 | Genentech Inc | ANTI-CD79b ANTIBODIES AND HUMANIZED IMMUNOCONJUGATES |
EP2626371A1 (en) | 2007-07-31 | 2013-08-14 | MedImmune, LLC | Multispecific epitope binding proteins and uses thereof |
EP2185188B1 (en) * | 2007-08-22 | 2014-08-06 | Medarex, L.L.C. | Site-specific attachment of drugs or other agents to engineered antibodies with c-terminal extensions |
GB2468232B (en) * | 2007-11-30 | 2012-10-24 | Glaxo Group Ltd | Antigen-bindng constructs |
US8454960B2 (en) | 2008-01-03 | 2013-06-04 | The Scripps Research Institute | Multispecific antibody targeting and multivalency through modular recognition domains |
EA021967B1 (en) | 2008-01-03 | 2015-10-30 | Дзе Скриппс Рисерч Инститьют | Antibody targeting through a modular recognition domain |
EA032828B1 (en) * | 2008-10-10 | 2019-07-31 | Аптево Рисёрч Энд Девелопмент Ллс | Tcr complex immunotherapeutics |
CA2889453C (en) * | 2009-03-20 | 2018-11-06 | Amgen Inc. | Carrier immunoglobulins and uses thereof |
ES2744226T3 (en) * | 2011-05-08 | 2020-02-24 | Legochem Biosciences Inc | Protein-active agent conjugates and method to prepare them |
CA2837169C (en) | 2011-05-24 | 2021-11-09 | Zyngenia, Inc. | Multispecific complexes comprising angiopoietin-2-binding peptide and their uses |
ES2667864T3 (en) * | 2011-06-22 | 2018-05-14 | F. Hoffmann-La Roche Ag | Removal of target cells by specific cytotoxic T lymphocytes from circulating viruses using complexes comprising MHC class I |
US9580509B2 (en) | 2011-11-07 | 2017-02-28 | Medimmune, Llc | Multispecific and multivalent binding proteins and uses thereof |
WO2013096829A2 (en) | 2011-12-22 | 2013-06-27 | Arizona Biomedical Research Commission | Activation of cellular assault processes in the treatment of glioblastoma multiforme |
MX2014010495A (en) * | 2012-03-02 | 2014-11-14 | Ablynx Nv | Pseudomonas aeruginosa pcrv binding single variable domain antibodies. |
EP2935589A1 (en) | 2012-12-18 | 2015-10-28 | Novartis AG | Compositions and methods that utilize a peptide tag that binds to hyaluronan |
-
2014
- 2014-12-23 AU AU2014373593A patent/AU2014373593B2/en active Active
- 2014-12-23 WO PCT/CA2014/051263 patent/WO2015095972A1/en active Application Filing
- 2014-12-23 CN CN201480075958.5A patent/CN106459981A/en active Pending
- 2014-12-23 JP JP2016543004A patent/JP6585600B2/en active Active
- 2014-12-23 KR KR1020167019769A patent/KR20160122127A/en active Search and Examination
- 2014-12-23 RU RU2016129724A patent/RU2016129724A/en unknown
- 2014-12-23 EP EP14874786.8A patent/EP3087185A4/en not_active Withdrawn
- 2014-12-23 US US15/107,884 patent/US11208497B2/en active Active
- 2014-12-23 BR BR112016014913A patent/BR112016014913A8/en not_active Application Discontinuation
- 2014-12-23 MX MX2016008355A patent/MX2016008355A/en unknown
- 2014-12-23 CA CA2934818A patent/CA2934818C/en active Active
- 2014-12-23 SG SG11201605093VA patent/SG11201605093VA/en unknown
-
2016
- 2016-06-21 IL IL246371A patent/IL246371A0/en unknown
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2012059882A2 (en) * | 2010-11-05 | 2012-05-10 | Rinat Neuroscience Corporation | Engineered polypeptide conjugates and methods for making thereof using transglutaminase |
Non-Patent Citations (4)
Title |
---|
GOLDMACHER, V.S. ET AL.: "Antibody-drug conjugates: using monoclonal antibodies for delivery of cytotoxic payloads to cancer cells", THERAPEUTIC DELIVERY, vol. 2, no. 3, March 2011 (2011-03-01), pages 397 - 416, XP055055352 * |
ORCUTT, K.D. ET AL.: "A modular IgG-scFv bispecific antibody topology", PROTEIN ENGINEERING, vol. 23, no. 4, 17 December 2009 (2009-12-17), pages 221 - 228, XP007914840 * |
See also references of EP3087185A4 * |
STROP, S. ET AL.: "Location matters: site of conjugation modulates stability and pharmacokinetics of antibody drug conjugates", CHEMISTRY AND BIOLOGY, vol. 20, no. 2, 21 February 2013 (2013-02-21), pages 161 - 167, XP055094948, Retrieved from the Internet <URL:http://dx.doi.org/10.1016/j.chembiol.2013.01.010> * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11834506B2 (en) | 2017-02-08 | 2023-12-05 | Dragonfly Therapeutics, Inc. | Multi-specific binding proteins that bind NKG2D, CD16, and a tumor-associated antigen for activation of natural killer cells and therapeutic uses thereof to treat cancer |
US11884732B2 (en) | 2017-02-20 | 2024-01-30 | Dragonfly Therapeutics, Inc. | Proteins binding HER2, NKG2D and CD16 |
US11884733B2 (en) | 2018-02-08 | 2024-01-30 | Dragonfly Therapeutics, Inc. | Antibody variable domains targeting the NKG2D receptor |
US11939384B1 (en) | 2018-02-08 | 2024-03-26 | Dragonfly Therapeutics, Inc. | Antibody variable domains targeting the NKG2D receptor |
Also Published As
Publication number | Publication date |
---|---|
AU2014373593B2 (en) | 2020-07-16 |
US11208497B2 (en) | 2021-12-28 |
US20170008970A1 (en) | 2017-01-12 |
MX2016008355A (en) | 2016-10-28 |
KR20160122127A (en) | 2016-10-21 |
AU2014373593A1 (en) | 2016-07-14 |
EP3087185A4 (en) | 2017-09-06 |
BR112016014913A8 (en) | 2020-06-09 |
JP2017501728A (en) | 2017-01-19 |
CA2934818A1 (en) | 2015-07-02 |
IL246371A0 (en) | 2016-08-31 |
JP6585600B2 (en) | 2019-10-02 |
CA2934818C (en) | 2022-05-17 |
SG11201605093VA (en) | 2016-07-28 |
BR112016014913A2 (en) | 2017-08-08 |
RU2016129724A3 (en) | 2018-10-18 |
EP3087185A1 (en) | 2016-11-02 |
RU2016129724A (en) | 2018-01-30 |
CN106459981A (en) | 2017-02-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU2014373593B2 (en) | Antibodies comprising C-terminal light chain polypeptide extensions and conjugates and methods of use thereof | |
US10517959B2 (en) | Binding protein drug conjugates comprising anthracycline derivatives | |
AU2015215015B2 (en) | Antibody-drug conjugates and immunotoxins | |
US10112999B2 (en) | Anti-PRLR antibody-drug conjugates (ADC) and uses thereof | |
TWI845724B (en) | Polypeptide complex for conjugation and use thereof | |
WO2022188743A1 (en) | Anti-her2 antibody-immune agonist conjugate and applications thereof | |
JP7547205B2 (en) | Anti-her2 biparatopic antibody-drug conjugates and methods of use | |
CN116115771A (en) | Materials and methods relating to linkers for use in protein drug conjugates | |
AU2020318112C1 (en) | Polypeptide complex for conjugation and use thereof | |
WO2023178451A1 (en) | Anti-folate receptor alpha antibodies and methods of use | |
WO2023141714A1 (en) | Methods of using anti-her2 biparatopic antibody-drug conjugates in the treatment of cancer |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 14874786 Country of ref document: EP Kind code of ref document: A1 |
|
REEP | Request for entry into the european phase |
Ref document number: 2014874786 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 246371 Country of ref document: IL Ref document number: 2014874786 Country of ref document: EP |
|
ENP | Entry into the national phase |
Ref document number: 2934818 Country of ref document: CA Ref document number: 2016543004 Country of ref document: JP Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: MX/A/2016/008355 Country of ref document: MX |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
WWE | Wipo information: entry into national phase |
Ref document number: 15107884 Country of ref document: US |
|
ENP | Entry into the national phase |
Ref document number: 2014373593 Country of ref document: AU Date of ref document: 20141223 Kind code of ref document: A |
|
ENP | Entry into the national phase |
Ref document number: 20167019769 Country of ref document: KR Kind code of ref document: A |
|
ENP | Entry into the national phase |
Ref document number: 2016129724 Country of ref document: RU Kind code of ref document: A |
|
REG | Reference to national code |
Ref country code: BR Ref legal event code: B01A Ref document number: 112016014913 Country of ref document: BR |
|
ENP | Entry into the national phase |
Ref document number: 112016014913 Country of ref document: BR Kind code of ref document: A2 Effective date: 20160623 |