CN114904015A - 包含抗cldn18.2的抗体或其抗原结合片段的抗体药物偶联物及其用途 - Google Patents
包含抗cldn18.2的抗体或其抗原结合片段的抗体药物偶联物及其用途 Download PDFInfo
- Publication number
- CN114904015A CN114904015A CN202210133787.3A CN202210133787A CN114904015A CN 114904015 A CN114904015 A CN 114904015A CN 202210133787 A CN202210133787 A CN 202210133787A CN 114904015 A CN114904015 A CN 114904015A
- Authority
- CN
- China
- Prior art keywords
- seq
- antibody
- cdr
- ser
- gly
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000000611 antibody drug conjugate Substances 0.000 title claims abstract description 232
- 229940049595 antibody-drug conjugate Drugs 0.000 title claims abstract description 224
- 239000012634 fragment Substances 0.000 title claims abstract description 56
- 230000027455 binding Effects 0.000 title claims abstract description 26
- 238000009739 binding Methods 0.000 title claims abstract description 24
- 239000000427 antigen Substances 0.000 title claims description 20
- 102000036639 antigens Human genes 0.000 title claims description 19
- 108091007433 antigens Proteins 0.000 title claims description 19
- 239000003814 drug Substances 0.000 claims abstract description 112
- 229940079593 drug Drugs 0.000 claims abstract description 107
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 12
- 238000002360 preparation method Methods 0.000 claims abstract description 12
- 238000000034 method Methods 0.000 claims description 88
- 206010028980 Neoplasm Diseases 0.000 claims description 69
- 230000002829 reductive effect Effects 0.000 claims description 31
- 238000005859 coupling reaction Methods 0.000 claims description 24
- 230000008878 coupling Effects 0.000 claims description 23
- 238000010168 coupling process Methods 0.000 claims description 23
- 241001529936 Murinae Species 0.000 claims description 18
- 201000011510 cancer Diseases 0.000 claims description 16
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 15
- 150000001875 compounds Chemical class 0.000 claims description 12
- 108010047041 Complementarity Determining Regions Proteins 0.000 claims description 10
- 229940044665 STING agonist Drugs 0.000 claims description 10
- 231100000433 cytotoxic Toxicity 0.000 claims description 7
- 230000001472 cytotoxic effect Effects 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- -1 camptothecin compound Chemical class 0.000 claims description 6
- KLWPJMFMVPTNCC-UHFFFAOYSA-N Camptothecin Natural products CCC1(O)C(=O)OCC2=C1C=C3C4Nc5ccccc5C=C4CN3C2=O KLWPJMFMVPTNCC-UHFFFAOYSA-N 0.000 claims description 5
- 239000007853 buffer solution Substances 0.000 claims description 5
- 229940127093 camptothecin Drugs 0.000 claims description 5
- 125000004122 cyclic group Chemical group 0.000 claims description 5
- 201000010099 disease Diseases 0.000 claims description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 5
- VSJKWCGYPAHWDS-UHFFFAOYSA-N dl-camptothecin Natural products C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)C5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-UHFFFAOYSA-N 0.000 claims description 5
- 230000001024 immunotherapeutic effect Effects 0.000 claims description 5
- 229940126586 small molecule drug Drugs 0.000 claims description 5
- FBFJOZZTIXSPPR-UHFFFAOYSA-N 1-(4-aminobutyl)-2-(ethoxymethyl)imidazo[4,5-c]quinolin-4-amine Chemical compound C1=CC=CC2=C(N(C(COCC)=N3)CCCCN)C3=C(N)N=C21 FBFJOZZTIXSPPR-UHFFFAOYSA-N 0.000 claims description 3
- 229940123780 DNA topoisomerase I inhibitor Drugs 0.000 claims description 3
- 229940124613 TLR 7/8 agonist Drugs 0.000 claims description 3
- 239000000365 Topoisomerase I Inhibitor Substances 0.000 claims description 3
- 229940122429 Tubulin inhibitor Drugs 0.000 claims description 3
- 108010044540 auristatin Proteins 0.000 claims description 3
- 239000003153 chemical reaction reagent Substances 0.000 claims description 3
- 239000003960 organic solvent Substances 0.000 claims description 3
- 239000004472 Lysine Substances 0.000 claims description 2
- 108010003723 Single-Domain Antibodies Proteins 0.000 claims description 2
- 238000004132 cross linking Methods 0.000 claims description 2
- 235000018417 cysteine Nutrition 0.000 claims description 2
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 229940124669 imidazoquinoline Drugs 0.000 claims description 2
- 239000011259 mixed solution Substances 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 33
- 230000001588 bifunctional effect Effects 0.000 claims 1
- 210000004027 cell Anatomy 0.000 description 98
- 239000000243 solution Substances 0.000 description 85
- 238000002474 experimental method Methods 0.000 description 67
- 150000001413 amino acids Chemical class 0.000 description 48
- 230000000694 effects Effects 0.000 description 38
- 230000014509 gene expression Effects 0.000 description 34
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 32
- 238000004458 analytical method Methods 0.000 description 31
- 230000009467 reduction Effects 0.000 description 30
- 108010010147 glycylglutamine Proteins 0.000 description 23
- 239000000523 sample Substances 0.000 description 22
- 241000699666 Mus <mouse, genus> Species 0.000 description 21
- 206010017758 gastric cancer Diseases 0.000 description 19
- 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 18
- PDIDTSZKKFEDMB-UWVGGRQHSA-N Lys-Pro-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O PDIDTSZKKFEDMB-UWVGGRQHSA-N 0.000 description 18
- WJZLEENECIOOSA-WDSKDSINSA-N Gly-Asn-Gln Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)O WJZLEENECIOOSA-WDSKDSINSA-N 0.000 description 17
- 108010008685 alanyl-glutamyl-aspartic acid Proteins 0.000 description 17
- 108010044292 tryptophyltyrosine Proteins 0.000 description 17
- TYYLDKGBCJGJGW-UHFFFAOYSA-N L-tryptophan-L-tyrosine Natural products C=1NC2=CC=CC=C2C=1CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 TYYLDKGBCJGJGW-UHFFFAOYSA-N 0.000 description 16
- 241000699670 Mus sp. Species 0.000 description 16
- 239000013642 negative control Substances 0.000 description 16
- 238000000746 purification Methods 0.000 description 16
- JXFLPKSDLDEOQK-JHEQGTHGSA-N Gln-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCC(N)=O JXFLPKSDLDEOQK-JHEQGTHGSA-N 0.000 description 15
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 15
- UIGMAMGZOJVTDN-WHFBIAKZSA-N Ser-Gly-Ser Chemical compound OC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O UIGMAMGZOJVTDN-WHFBIAKZSA-N 0.000 description 15
- 208000005718 Stomach Neoplasms Diseases 0.000 description 15
- 108010037850 glycylvaline Proteins 0.000 description 15
- ZDILXFDENZVOTL-BPNCWPANSA-N Ala-Val-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZDILXFDENZVOTL-BPNCWPANSA-N 0.000 description 14
- NVTPVQLIZCOJFK-FOHZUACHSA-N Gly-Thr-Asp Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O NVTPVQLIZCOJFK-FOHZUACHSA-N 0.000 description 14
- KLYYKKGCPOGDPE-OEAJRASXSA-N Phe-Thr-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O KLYYKKGCPOGDPE-OEAJRASXSA-N 0.000 description 14
- QOIKZODVIPOPDD-AVGNSLFASA-N Tyr-Cys-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(O)=O QOIKZODVIPOPDD-AVGNSLFASA-N 0.000 description 14
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 14
- 108010050848 glycylleucine Proteins 0.000 description 14
- 201000011549 stomach cancer Diseases 0.000 description 14
- KYQNAIMCTRZLNP-QSFUFRPTSA-N Asp-Ile-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O KYQNAIMCTRZLNP-QSFUFRPTSA-N 0.000 description 13
- NIXHTNJAGGFBAW-CIUDSAMLSA-N Cys-Lys-Ser Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CS)N NIXHTNJAGGFBAW-CIUDSAMLSA-N 0.000 description 13
- RFTVTKBHDXCEEX-WDSKDSINSA-N Glu-Ser-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RFTVTKBHDXCEEX-WDSKDSINSA-N 0.000 description 13
- BYYNJRSNDARRBX-YFKPBYRVSA-N Gly-Gln-Gly Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O BYYNJRSNDARRBX-YFKPBYRVSA-N 0.000 description 13
- SCWYHUQOOFRVHP-MBLNEYKQSA-N Gly-Ile-Thr Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SCWYHUQOOFRVHP-MBLNEYKQSA-N 0.000 description 13
- RNYLNYTYMXACRI-VFAJRCTISA-N Leu-Thr-Trp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O RNYLNYTYMXACRI-VFAJRCTISA-N 0.000 description 13
- JARJPEMLQAWNBR-GUBZILKMSA-N Pro-Asp-Arg Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O JARJPEMLQAWNBR-GUBZILKMSA-N 0.000 description 13
- PCWLNNZTBJTZRN-AVGNSLFASA-N Pro-Pro-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 PCWLNNZTBJTZRN-AVGNSLFASA-N 0.000 description 13
- RXUOAOOZIWABBW-XGEHTFHBSA-N Ser-Thr-Arg Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N RXUOAOOZIWABBW-XGEHTFHBSA-N 0.000 description 13
- BIBYEFRASCNLAA-CDMKHQONSA-N Thr-Phe-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CC=CC=C1 BIBYEFRASCNLAA-CDMKHQONSA-N 0.000 description 13
- QUILOGWWLXMSAT-IHRRRGAJSA-N Tyr-Gln-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O QUILOGWWLXMSAT-IHRRRGAJSA-N 0.000 description 13
- ZYVAAYAOTVJBSS-GMVOTWDCSA-N Tyr-Trp-Ala Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C)C(O)=O ZYVAAYAOTVJBSS-GMVOTWDCSA-N 0.000 description 13
- 108010081404 acein-2 Proteins 0.000 description 13
- 108010013835 arginine glutamate Proteins 0.000 description 13
- PXKACEXYLPBMAD-JBDRJPRFSA-N Ile-Ser-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)O)N PXKACEXYLPBMAD-JBDRJPRFSA-N 0.000 description 12
- SKRGVGLIRUGANF-AVGNSLFASA-N Lys-Leu-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SKRGVGLIRUGANF-AVGNSLFASA-N 0.000 description 12
- BIWVVOHTKDLRMP-ULQDDVLXSA-N Tyr-Pro-Leu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O BIWVVOHTKDLRMP-ULQDDVLXSA-N 0.000 description 12
- 230000021615 conjugation Effects 0.000 description 12
- FSXRLASFHBWESK-UHFFFAOYSA-N dipeptide phenylalanyl-tyrosine Natural products C=1C=C(O)C=CC=1CC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FSXRLASFHBWESK-UHFFFAOYSA-N 0.000 description 12
- 238000000338 in vitro Methods 0.000 description 12
- FEHQLKKBVJHSEC-SZMVWBNQSA-N Leu-Glu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC(C)C)C(O)=O)=CNC2=C1 FEHQLKKBVJHSEC-SZMVWBNQSA-N 0.000 description 11
- 101710196623 Stimulator of interferon genes protein Proteins 0.000 description 11
- 238000003556 assay Methods 0.000 description 11
- 238000011156 evaluation Methods 0.000 description 11
- 238000004128 high performance liquid chromatography Methods 0.000 description 11
- 230000005764 inhibitory process Effects 0.000 description 11
- AVZHGSCDKIQZPQ-CIUDSAMLSA-N Glu-Arg-Ala Chemical compound C[C@H](NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](N)CCC(O)=O)C(O)=O AVZHGSCDKIQZPQ-CIUDSAMLSA-N 0.000 description 10
- PMGDADKJMCOXHX-UHFFFAOYSA-N L-Arginyl-L-glutamin-acetat Natural products NC(=N)NCCCC(N)C(=O)NC(CCC(N)=O)C(O)=O PMGDADKJMCOXHX-UHFFFAOYSA-N 0.000 description 10
- VQPPIMUZCZCOIL-GUBZILKMSA-N Leu-Gln-Ala Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O VQPPIMUZCZCOIL-GUBZILKMSA-N 0.000 description 10
- BPCLGWHVPVTTFM-QWRGUYRKSA-N Phe-Ser-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)NCC(O)=O BPCLGWHVPVTTFM-QWRGUYRKSA-N 0.000 description 10
- FUMGHWDRRFCKEP-CIUDSAMLSA-N Ser-Leu-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O FUMGHWDRRFCKEP-CIUDSAMLSA-N 0.000 description 10
- PJIQEIFXZPCWOJ-FXQIFTODSA-N Ser-Pro-Asp Chemical compound [H]N[C@@H](CO)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O PJIQEIFXZPCWOJ-FXQIFTODSA-N 0.000 description 10
- PAXANSWUSVPFNK-IUKAMOBKSA-N Thr-Ile-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H]([C@@H](C)O)N PAXANSWUSVPFNK-IUKAMOBKSA-N 0.000 description 10
- MBLJBGZWLHTJBH-SZMVWBNQSA-N Trp-Val-Arg Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O)=CNC2=C1 MBLJBGZWLHTJBH-SZMVWBNQSA-N 0.000 description 10
- VHIZXDZMTDVFGX-DCAQKATOSA-N Val-Ser-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](C(C)C)N VHIZXDZMTDVFGX-DCAQKATOSA-N 0.000 description 10
- 108010008355 arginyl-glutamine Proteins 0.000 description 10
- 238000004191 hydrophobic interaction chromatography Methods 0.000 description 10
- 238000005259 measurement Methods 0.000 description 10
- 238000007920 subcutaneous administration Methods 0.000 description 10
- 239000000126 substance Substances 0.000 description 10
- OTUQSEPIIVBYEM-IHRRRGAJSA-N Arg-Asn-Tyr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O OTUQSEPIIVBYEM-IHRRRGAJSA-N 0.000 description 9
- OYTPNWYZORARHL-XHNCKOQMSA-N Gln-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)N)N OYTPNWYZORARHL-XHNCKOQMSA-N 0.000 description 9
- JVSBYEDSSRZQGV-GUBZILKMSA-N Glu-Asp-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CCC(O)=O JVSBYEDSSRZQGV-GUBZILKMSA-N 0.000 description 9
- 241000880493 Leptailurus serval Species 0.000 description 9
- IAJFFZORSWOZPQ-SRVKXCTJSA-N Leu-Leu-Asn Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O IAJFFZORSWOZPQ-SRVKXCTJSA-N 0.000 description 9
- SPSSJSICDYYTQN-HJGDQZAQSA-N Met-Thr-Gln Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCC(N)=O SPSSJSICDYYTQN-HJGDQZAQSA-N 0.000 description 9
- QYSFWUIXDFJUDW-DCAQKATOSA-N Ser-Leu-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O QYSFWUIXDFJUDW-DCAQKATOSA-N 0.000 description 9
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 9
- 230000004540 complement-dependent cytotoxicity Effects 0.000 description 9
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 9
- 230000002147 killing effect Effects 0.000 description 9
- 108090000623 proteins and genes Proteins 0.000 description 9
- PXHCFKXNSBJSTQ-KKUMJFAQSA-N Lys-Asn-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCCCN)N)O PXHCFKXNSBJSTQ-KKUMJFAQSA-N 0.000 description 8
- FMDHKPRACUXATF-ACZMJKKPSA-N Ser-Gln-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O FMDHKPRACUXATF-ACZMJKKPSA-N 0.000 description 8
- LGIMRDKGABDMBN-DCAQKATOSA-N Ser-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CO)N LGIMRDKGABDMBN-DCAQKATOSA-N 0.000 description 8
- SZEIFUXUTBBQFQ-STQMWFEESA-N Tyr-Pro-Gly Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O SZEIFUXUTBBQFQ-STQMWFEESA-N 0.000 description 8
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 8
- 210000004408 hybridoma Anatomy 0.000 description 8
- 210000002865 immune cell Anatomy 0.000 description 8
- 108010073969 valyllysine Proteins 0.000 description 8
- QUCCLIXMVPIVOB-BZSNNMDCSA-N Asn-Tyr-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CC(=O)N)N QUCCLIXMVPIVOB-BZSNNMDCSA-N 0.000 description 7
- ZYRXTRTUCAVNBQ-GVXVVHGQSA-N Glu-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N ZYRXTRTUCAVNBQ-GVXVVHGQSA-N 0.000 description 7
- BPQYBFAXRGMGGY-LAEOZQHASA-N Gly-Gln-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)CN BPQYBFAXRGMGGY-LAEOZQHASA-N 0.000 description 7
- UIQGJYUEQDOODF-KWQFWETISA-N Gly-Tyr-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)CN)CC1=CC=C(O)C=C1 UIQGJYUEQDOODF-KWQFWETISA-N 0.000 description 7
- 101000800483 Homo sapiens Toll-like receptor 8 Proteins 0.000 description 7
- ZLFNNVATRMCAKN-ZKWXMUAHSA-N Ile-Ser-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)NCC(=O)O)N ZLFNNVATRMCAKN-ZKWXMUAHSA-N 0.000 description 7
- FAELBUXXFQLUAX-AJNGGQMLSA-N Leu-Leu-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(C)C FAELBUXXFQLUAX-AJNGGQMLSA-N 0.000 description 7
- PPGBXYKMUMHFBF-KATARQTJSA-N Leu-Ser-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PPGBXYKMUMHFBF-KATARQTJSA-N 0.000 description 7
- UWKNTTJNVSYXPC-CIUDSAMLSA-N Lys-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN UWKNTTJNVSYXPC-CIUDSAMLSA-N 0.000 description 7
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 7
- YMIZSYUAZJSOFL-SRVKXCTJSA-N Phe-Ser-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O YMIZSYUAZJSOFL-SRVKXCTJSA-N 0.000 description 7
- UQFYNFTYDHUIMI-WHFBIAKZSA-N Ser-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H](N)CO UQFYNFTYDHUIMI-WHFBIAKZSA-N 0.000 description 7
- WSTIOCFMWXNOCX-YUMQZZPRSA-N Ser-Gly-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CO)N WSTIOCFMWXNOCX-YUMQZZPRSA-N 0.000 description 7
- 229940124614 TLR 8 agonist Drugs 0.000 description 7
- KKPOGALELPLJTL-MEYUZBJRSA-N Thr-Lys-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 KKPOGALELPLJTL-MEYUZBJRSA-N 0.000 description 7
- 102100033110 Toll-like receptor 8 Human genes 0.000 description 7
- PZTZYZUTCPZWJH-FXQIFTODSA-N Val-Ser-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)O)N PZTZYZUTCPZWJH-FXQIFTODSA-N 0.000 description 7
- 238000002835 absorbance Methods 0.000 description 7
- 235000001014 amino acid Nutrition 0.000 description 7
- 229940024606 amino acid Drugs 0.000 description 7
- 230000010056 antibody-dependent cellular cytotoxicity Effects 0.000 description 7
- 230000022534 cell killing Effects 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 7
- 108010031719 prolyl-serine Proteins 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 238000004366 reverse phase liquid chromatography Methods 0.000 description 7
- 239000006228 supernatant Substances 0.000 description 7
- 210000004881 tumor cell Anatomy 0.000 description 7
- HTSSXFASOUSJQG-IHPCNDPISA-N Asp-Tyr-Trp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O HTSSXFASOUSJQG-IHPCNDPISA-N 0.000 description 6
- RXGLHDWAZQECBI-SRVKXCTJSA-N Leu-Leu-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O RXGLHDWAZQECBI-SRVKXCTJSA-N 0.000 description 6
- KIZIOFNVSOSKJI-CIUDSAMLSA-N Leu-Ser-Cys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O)N KIZIOFNVSOSKJI-CIUDSAMLSA-N 0.000 description 6
- WVJNGSFKBKOKRV-AJNGGQMLSA-N Lys-Leu-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WVJNGSFKBKOKRV-AJNGGQMLSA-N 0.000 description 6
- DIBZLYZXTSVGLN-CIUDSAMLSA-N Lys-Ser-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O DIBZLYZXTSVGLN-CIUDSAMLSA-N 0.000 description 6
- CGBYDGAJHSOGFQ-LPEHRKFASA-N Pro-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@@H]2CCCN2 CGBYDGAJHSOGFQ-LPEHRKFASA-N 0.000 description 6
- IHCXPSYCHXFXKT-DCAQKATOSA-N Pro-Arg-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O IHCXPSYCHXFXKT-DCAQKATOSA-N 0.000 description 6
- YMTLKLXDFCSCNX-BYPYZUCNSA-N Ser-Gly-Gly Chemical compound OC[C@H](N)C(=O)NCC(=O)NCC(O)=O YMTLKLXDFCSCNX-BYPYZUCNSA-N 0.000 description 6
- BMKNXTJLHFIAAH-CIUDSAMLSA-N Ser-Ser-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O BMKNXTJLHFIAAH-CIUDSAMLSA-N 0.000 description 6
- FQPDRTDDEZXCEC-SVSWQMSJSA-N Thr-Ile-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O FQPDRTDDEZXCEC-SVSWQMSJSA-N 0.000 description 6
- 108010092854 aspartyllysine Proteins 0.000 description 6
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 6
- 229940127121 immunoconjugate Drugs 0.000 description 6
- 238000011534 incubation Methods 0.000 description 6
- 108010054155 lysyllysine Proteins 0.000 description 6
- 230000000770 proinflammatory effect Effects 0.000 description 6
- 239000011535 reaction buffer Substances 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
- 238000002054 transplantation Methods 0.000 description 6
- 108010038745 tryptophylglycine Proteins 0.000 description 6
- 239000003643 water by type Substances 0.000 description 6
- YWWATNIVMOCSAV-UBHSHLNASA-N Ala-Arg-Phe Chemical compound NC(=N)NCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 YWWATNIVMOCSAV-UBHSHLNASA-N 0.000 description 5
- LBYMZCVBOKYZNS-CIUDSAMLSA-N Ala-Leu-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O LBYMZCVBOKYZNS-CIUDSAMLSA-N 0.000 description 5
- AWNAEZICPNGAJK-FXQIFTODSA-N Ala-Met-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CO)C(O)=O AWNAEZICPNGAJK-FXQIFTODSA-N 0.000 description 5
- QBQVKUNBCAFXSV-ULQDDVLXSA-N Arg-Lys-Tyr Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 QBQVKUNBCAFXSV-ULQDDVLXSA-N 0.000 description 5
- SYZWMVSXBZCOBZ-QXEWZRGKSA-N Asn-Val-Met Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CC(=O)N)N SYZWMVSXBZCOBZ-QXEWZRGKSA-N 0.000 description 5
- JSHWXQIZOCVWIA-ZKWXMUAHSA-N Asp-Ser-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O JSHWXQIZOCVWIA-ZKWXMUAHSA-N 0.000 description 5
- MNQMTYSEKZHIDF-GCJQMDKQSA-N Asp-Thr-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O MNQMTYSEKZHIDF-GCJQMDKQSA-N 0.000 description 5
- ZFBBMCKQSNJZSN-AUTRQRHGSA-N Gln-Val-Gln Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O ZFBBMCKQSNJZSN-AUTRQRHGSA-N 0.000 description 5
- YQPFCZVKMUVZIN-AUTRQRHGSA-N Glu-Val-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O YQPFCZVKMUVZIN-AUTRQRHGSA-N 0.000 description 5
- MIIVFRCYJABHTQ-ONGXEEELSA-N Gly-Leu-Val Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O MIIVFRCYJABHTQ-ONGXEEELSA-N 0.000 description 5
- DNAZKGFYFRGZIH-QWRGUYRKSA-N Gly-Tyr-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=C(O)C=C1 DNAZKGFYFRGZIH-QWRGUYRKSA-N 0.000 description 5
- VBOFRJNDIOPNDO-YUMQZZPRSA-N His-Gly-Asn Chemical compound C1=C(NC=N1)C[C@@H](C(=O)NCC(=O)N[C@@H](CC(=O)N)C(=O)O)N VBOFRJNDIOPNDO-YUMQZZPRSA-N 0.000 description 5
- BRTVHXHCUSXYRI-CIUDSAMLSA-N Leu-Ser-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O BRTVHXHCUSXYRI-CIUDSAMLSA-N 0.000 description 5
- LCNASHSOFMRYFO-WDCWCFNPSA-N Leu-Thr-Gln Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCC(N)=O LCNASHSOFMRYFO-WDCWCFNPSA-N 0.000 description 5
- VUBIPAHVHMZHCM-KKUMJFAQSA-N Leu-Tyr-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CO)C(O)=O)CC1=CC=C(O)C=C1 VUBIPAHVHMZHCM-KKUMJFAQSA-N 0.000 description 5
- AIMGJYMCTAABEN-GVXVVHGQSA-N Leu-Val-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O AIMGJYMCTAABEN-GVXVVHGQSA-N 0.000 description 5
- QESXLSQLQHHTIX-RHYQMDGZSA-N Leu-Val-Thr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QESXLSQLQHHTIX-RHYQMDGZSA-N 0.000 description 5
- VTFXTWDFPTWNJY-RHYQMDGZSA-N Pro-Leu-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VTFXTWDFPTWNJY-RHYQMDGZSA-N 0.000 description 5
- 239000012980 RPMI-1640 medium Substances 0.000 description 5
- KJMOINFQVCCSDX-XKBZYTNZSA-N Ser-Gln-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KJMOINFQVCCSDX-XKBZYTNZSA-N 0.000 description 5
- HNDMFDBQXYZSRM-IHRRRGAJSA-N Ser-Val-Phe Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O HNDMFDBQXYZSRM-IHRRRGAJSA-N 0.000 description 5
- NJEMRSFGDNECGF-GCJQMDKQSA-N Thr-Ala-Asp Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O NJEMRSFGDNECGF-GCJQMDKQSA-N 0.000 description 5
- RCMHSGRBJCMFLR-BPUTZDHNSA-N Trp-Met-Asn Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(O)=O)=CNC2=C1 RCMHSGRBJCMFLR-BPUTZDHNSA-N 0.000 description 5
- NUQZCPSZHGIYTA-HKUYNNGSSA-N Tyr-Trp-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CC3=CC=C(C=C3)O)N NUQZCPSZHGIYTA-HKUYNNGSSA-N 0.000 description 5
- ANHVRCNNGJMJNG-BZSNNMDCSA-N Tyr-Tyr-Cys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)N[C@@H](CS)C(=O)O)N)O ANHVRCNNGJMJNG-BZSNNMDCSA-N 0.000 description 5
- WYOBRXPIZVKNMF-IRXDYDNUSA-N Tyr-Tyr-Gly Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)NCC(O)=O)C1=CC=C(O)C=C1 WYOBRXPIZVKNMF-IRXDYDNUSA-N 0.000 description 5
- HZDQUVQEVVYDDA-ACRUOGEOSA-N Tyr-Tyr-Leu Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 HZDQUVQEVVYDDA-ACRUOGEOSA-N 0.000 description 5
- JQTYTBPCSOAZHI-FXQIFTODSA-N Val-Ser-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O)N JQTYTBPCSOAZHI-FXQIFTODSA-N 0.000 description 5
- HTONZBWRYUKUKC-RCWTZXSCSA-N Val-Thr-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O HTONZBWRYUKUKC-RCWTZXSCSA-N 0.000 description 5
- OWFGFHQMSBTKLX-UFYCRDLUSA-N Val-Tyr-Tyr Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N OWFGFHQMSBTKLX-UFYCRDLUSA-N 0.000 description 5
- 108010068265 aspartyltyrosine Proteins 0.000 description 5
- 230000012202 endocytosis Effects 0.000 description 5
- 208000010749 gastric carcinoma Diseases 0.000 description 5
- 108010067216 glycyl-glycyl-glycine Proteins 0.000 description 5
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 5
- 108010017391 lysylvaline Proteins 0.000 description 5
- 229950001460 sacituzumab Drugs 0.000 description 5
- 201000000498 stomach carcinoma Diseases 0.000 description 5
- 230000008685 targeting Effects 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- 108010020532 tyrosyl-proline Proteins 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- LMFAFFIRCNUJJO-UHFFFAOYSA-N 5-(4-azidophenyl)-8-iodo-3-methyl-1,2,4,5-tetrahydro-3-benzazepin-7-ol Chemical compound C1N(C)CCC2=CC(I)=C(O)C=C2C1C1=CC=C(N=[N+]=[N-])C=C1 LMFAFFIRCNUJJO-UHFFFAOYSA-N 0.000 description 4
- KXEVYGKATAMXJJ-ACZMJKKPSA-N Ala-Glu-Asp Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O KXEVYGKATAMXJJ-ACZMJKKPSA-N 0.000 description 4
- NBTGEURICRTMGL-WHFBIAKZSA-N Ala-Gly-Ser Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O NBTGEURICRTMGL-WHFBIAKZSA-N 0.000 description 4
- AJBVYEYZVYPFCF-CIUDSAMLSA-N Ala-Lys-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O AJBVYEYZVYPFCF-CIUDSAMLSA-N 0.000 description 4
- 101100330725 Arabidopsis thaliana DAR4 gene Proteins 0.000 description 4
- PQWTZSNVWSOFFK-FXQIFTODSA-N Arg-Asp-Asn Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)CN=C(N)N PQWTZSNVWSOFFK-FXQIFTODSA-N 0.000 description 4
- ISJWBVIYRBAXEB-CIUDSAMLSA-N Arg-Ser-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O ISJWBVIYRBAXEB-CIUDSAMLSA-N 0.000 description 4
- IICZCLFBILYRCU-WHFBIAKZSA-N Asn-Gly-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IICZCLFBILYRCU-WHFBIAKZSA-N 0.000 description 4
- JSNWZMFSLIWAHS-HJGDQZAQSA-N Asp-Thr-Leu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)O)NC(=O)[C@H](CC(=O)O)N)O JSNWZMFSLIWAHS-HJGDQZAQSA-N 0.000 description 4
- OHLLDUNVMPPUMD-DCAQKATOSA-N Cys-Leu-Val Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CS)N OHLLDUNVMPPUMD-DCAQKATOSA-N 0.000 description 4
- FQCILXROGNOZON-YUMQZZPRSA-N Gln-Pro-Gly Chemical compound NC(=O)CC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O FQCILXROGNOZON-YUMQZZPRSA-N 0.000 description 4
- WATXSTJXNBOHKD-LAEOZQHASA-N Glu-Asp-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O WATXSTJXNBOHKD-LAEOZQHASA-N 0.000 description 4
- KRRMJKMGWWXWDW-STQMWFEESA-N Gly-Arg-Phe Chemical compound NC(=N)NCCC[C@H](NC(=O)CN)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 KRRMJKMGWWXWDW-STQMWFEESA-N 0.000 description 4
- PDUHNKAFQXQNLH-ZETCQYMHSA-N Gly-Lys-Gly Chemical compound NCCCC[C@H](NC(=O)CN)C(=O)NCC(O)=O PDUHNKAFQXQNLH-ZETCQYMHSA-N 0.000 description 4
- 101001011382 Homo sapiens Interferon regulatory factor 3 Proteins 0.000 description 4
- 102100029843 Interferon regulatory factor 3 Human genes 0.000 description 4
- MVJRBCJCRYGCKV-GVXVVHGQSA-N Leu-Val-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O MVJRBCJCRYGCKV-GVXVVHGQSA-N 0.000 description 4
- SJDQOYTYNGZZJX-SRVKXCTJSA-N Met-Glu-Leu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O SJDQOYTYNGZZJX-SRVKXCTJSA-N 0.000 description 4
- 108010079364 N-glycylalanine Proteins 0.000 description 4
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 4
- 229920002873 Polyethylenimine Polymers 0.000 description 4
- YDTUEBLEAVANFH-RCWTZXSCSA-N Pro-Val-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 YDTUEBLEAVANFH-RCWTZXSCSA-N 0.000 description 4
- MUARUIBTKQJKFY-WHFBIAKZSA-N Ser-Gly-Asp Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O MUARUIBTKQJKFY-WHFBIAKZSA-N 0.000 description 4
- MUJQWSAWLLRJCE-KATARQTJSA-N Ser-Leu-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MUJQWSAWLLRJCE-KATARQTJSA-N 0.000 description 4
- AZWNCEBQZXELEZ-FXQIFTODSA-N Ser-Pro-Ser Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O AZWNCEBQZXELEZ-FXQIFTODSA-N 0.000 description 4
- JCLAFVNDBJMLBC-JBDRJPRFSA-N Ser-Ser-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JCLAFVNDBJMLBC-JBDRJPRFSA-N 0.000 description 4
- XJDMUQCLVSCRSJ-VZFHVOOUSA-N Ser-Thr-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O XJDMUQCLVSCRSJ-VZFHVOOUSA-N 0.000 description 4
- XSLXHSYIVPGEER-KZVJFYERSA-N Thr-Ala-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O XSLXHSYIVPGEER-KZVJFYERSA-N 0.000 description 4
- BMGOFDMKDVVGJG-NHCYSSNCSA-N Val-Asp-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N BMGOFDMKDVVGJG-NHCYSSNCSA-N 0.000 description 4
- WUFHZIRMAZZWRS-OSUNSFLBSA-N Val-Thr-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](C(C)C)N WUFHZIRMAZZWRS-OSUNSFLBSA-N 0.000 description 4
- 230000004913 activation Effects 0.000 description 4
- 108010024078 alanyl-glycyl-serine Proteins 0.000 description 4
- 229940125644 antibody drug Drugs 0.000 description 4
- 108010052670 arginyl-glutamyl-glutamic acid Proteins 0.000 description 4
- 108010047857 aspartylglycine Proteins 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 4
- 108010060199 cysteinylproline Proteins 0.000 description 4
- 239000013604 expression vector Substances 0.000 description 4
- 239000012091 fetal bovine serum Substances 0.000 description 4
- 238000000684 flow cytometry Methods 0.000 description 4
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 4
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 4
- 210000004962 mammalian cell Anatomy 0.000 description 4
- 201000002528 pancreatic cancer Diseases 0.000 description 4
- 208000008443 pancreatic carcinoma Diseases 0.000 description 4
- 108010020755 prolyl-glycyl-glycine Proteins 0.000 description 4
- 108010077112 prolyl-proline Proteins 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 108010069117 seryl-lysyl-aspartic acid Proteins 0.000 description 4
- 230000000638 stimulation Effects 0.000 description 4
- 238000002626 targeted therapy Methods 0.000 description 4
- 230000001225 therapeutic effect Effects 0.000 description 4
- 230000004614 tumor growth Effects 0.000 description 4
- 108010009962 valyltyrosine Proteins 0.000 description 4
- 239000013598 vector Substances 0.000 description 4
- SVUOLADPCWQTTE-UHFFFAOYSA-N 1h-1,2-benzodiazepine Chemical compound N1N=CC=CC2=CC=CC=C12 SVUOLADPCWQTTE-UHFFFAOYSA-N 0.000 description 3
- MNZHHDPWDWQJCQ-YUMQZZPRSA-N Ala-Leu-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O MNZHHDPWDWQJCQ-YUMQZZPRSA-N 0.000 description 3
- WQKAQKZRDIZYNV-VZFHVOOUSA-N Ala-Ser-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O WQKAQKZRDIZYNV-VZFHVOOUSA-N 0.000 description 3
- HGKHPCFTRQDHCU-IUCAKERBSA-N Arg-Pro-Gly Chemical compound NC(N)=NCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O HGKHPCFTRQDHCU-IUCAKERBSA-N 0.000 description 3
- SRUUBQBAVNQZGJ-LAEOZQHASA-N Asn-Gln-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC(=O)N)N SRUUBQBAVNQZGJ-LAEOZQHASA-N 0.000 description 3
- HNXWVVHIGTZTBO-LKXGYXEUSA-N Asn-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O HNXWVVHIGTZTBO-LKXGYXEUSA-N 0.000 description 3
- WLVLIYYBPPONRJ-GCJQMDKQSA-N Asn-Thr-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O WLVLIYYBPPONRJ-GCJQMDKQSA-N 0.000 description 3
- QNNBHTFDFFFHGC-KKUMJFAQSA-N Asn-Tyr-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O QNNBHTFDFFFHGC-KKUMJFAQSA-N 0.000 description 3
- MJIJBEYEHBKTIM-BYULHYEWSA-N Asn-Val-Asn Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N MJIJBEYEHBKTIM-BYULHYEWSA-N 0.000 description 3
- 241000283707 Capra Species 0.000 description 3
- NDNZRWUDUMTITL-FXQIFTODSA-N Cys-Ser-Val Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O NDNZRWUDUMTITL-FXQIFTODSA-N 0.000 description 3
- 102000003915 DNA Topoisomerases Human genes 0.000 description 3
- 108090000323 DNA Topoisomerases Proteins 0.000 description 3
- 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 3
- 208000000461 Esophageal Neoplasms Diseases 0.000 description 3
- ZEEPYMXTJWIMSN-GUBZILKMSA-N Gln-Lys-Ser Chemical compound NCCCC[C@@H](C(=O)N[C@@H](CO)C(O)=O)NC(=O)[C@@H](N)CCC(N)=O ZEEPYMXTJWIMSN-GUBZILKMSA-N 0.000 description 3
- MSHXWFKYXJTLEZ-CIUDSAMLSA-N Gln-Met-Asn Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)N)N MSHXWFKYXJTLEZ-CIUDSAMLSA-N 0.000 description 3
- CKOFNWCLWRYUHK-XHNCKOQMSA-N Glu-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCC(=O)O)N)C(=O)O CKOFNWCLWRYUHK-XHNCKOQMSA-N 0.000 description 3
- HUFCEIHAFNVSNR-IHRRRGAJSA-N Glu-Gln-Tyr Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 HUFCEIHAFNVSNR-IHRRRGAJSA-N 0.000 description 3
- MWMJCGBSIORNCD-AVGNSLFASA-N Glu-Leu-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O MWMJCGBSIORNCD-AVGNSLFASA-N 0.000 description 3
- UQJNXZSSGQIPIQ-FBCQKBJTSA-N Gly-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)CN UQJNXZSSGQIPIQ-FBCQKBJTSA-N 0.000 description 3
- SYOJVRNQCXYEOV-XVKPBYJWSA-N Gly-Val-Glu Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SYOJVRNQCXYEOV-XVKPBYJWSA-N 0.000 description 3
- FULZDMOZUZKGQU-ONGXEEELSA-N Gly-Val-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)CN FULZDMOZUZKGQU-ONGXEEELSA-N 0.000 description 3
- 101000618467 Hypocrea jecorina (strain ATCC 56765 / BCRC 32924 / NRRL 11460 / Rut C-30) Endo-1,4-beta-xylanase 2 Proteins 0.000 description 3
- KAFOIVJDVSZUMD-DCAQKATOSA-N Leu-Gln-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O KAFOIVJDVSZUMD-DCAQKATOSA-N 0.000 description 3
- KAFOIVJDVSZUMD-UHFFFAOYSA-N Leu-Gln-Gln Natural products CC(C)CC(N)C(=O)NC(CCC(N)=O)C(=O)NC(CCC(N)=O)C(O)=O KAFOIVJDVSZUMD-UHFFFAOYSA-N 0.000 description 3
- XOWMDXHFSBCAKQ-SRVKXCTJSA-N Leu-Ser-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC(C)C XOWMDXHFSBCAKQ-SRVKXCTJSA-N 0.000 description 3
- MPGHETGWWWUHPY-CIUDSAMLSA-N Lys-Ala-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN MPGHETGWWWUHPY-CIUDSAMLSA-N 0.000 description 3
- KCXUCYYZNZFGLL-SRVKXCTJSA-N Lys-Ala-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O KCXUCYYZNZFGLL-SRVKXCTJSA-N 0.000 description 3
- NTBFKPBULZGXQL-KKUMJFAQSA-N Lys-Asp-Tyr Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 NTBFKPBULZGXQL-KKUMJFAQSA-N 0.000 description 3
- RFQATBGBLDAKGI-VHSXEESVSA-N Lys-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CCCCN)N)C(=O)O RFQATBGBLDAKGI-VHSXEESVSA-N 0.000 description 3
- LUTDBHBIHHREDC-IHRRRGAJSA-N Lys-Pro-Lys Chemical compound NCCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(O)=O LUTDBHBIHHREDC-IHRRRGAJSA-N 0.000 description 3
- MIFFFXHMAHFACR-KATARQTJSA-N Lys-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CCCCN MIFFFXHMAHFACR-KATARQTJSA-N 0.000 description 3
- FXHOOIRPVKKKFG-UHFFFAOYSA-N N,N-Dimethylacetamide Chemical compound CN(C)C(C)=O FXHOOIRPVKKKFG-UHFFFAOYSA-N 0.000 description 3
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 3
- 206010030155 Oesophageal carcinoma Diseases 0.000 description 3
- 206010033128 Ovarian cancer Diseases 0.000 description 3
- 206010061535 Ovarian neoplasm Diseases 0.000 description 3
- JOXIIFVCSATTDH-IHPCNDPISA-N Phe-Asn-Trp Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)O)N JOXIIFVCSATTDH-IHPCNDPISA-N 0.000 description 3
- QARPMYDMYVLFMW-KKUMJFAQSA-N Phe-Pro-Glu Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(O)=O)C1=CC=CC=C1 QARPMYDMYVLFMW-KKUMJFAQSA-N 0.000 description 3
- BSKMOCNNLNDIMU-CDMKHQONSA-N Phe-Thr-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O BSKMOCNNLNDIMU-CDMKHQONSA-N 0.000 description 3
- SJRQWEDYTKYHHL-SLFFLAALSA-N Phe-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CC3=CC=CC=C3)N)C(=O)O SJRQWEDYTKYHHL-SLFFLAALSA-N 0.000 description 3
- MHHQQZIFLWFZGR-DCAQKATOSA-N Pro-Lys-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O MHHQQZIFLWFZGR-DCAQKATOSA-N 0.000 description 3
- FDMKYQQYJKYCLV-GUBZILKMSA-N Pro-Pro-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 FDMKYQQYJKYCLV-GUBZILKMSA-N 0.000 description 3
- SRSPTFBENMJHMR-WHFBIAKZSA-N Ser-Ser-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SRSPTFBENMJHMR-WHFBIAKZSA-N 0.000 description 3
- SNXUIBACCONSOH-BWBBJGPYSA-N Ser-Thr-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CO)C(O)=O SNXUIBACCONSOH-BWBBJGPYSA-N 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- JMBRNXUOLJFURW-BEAPCOKYSA-N Thr-Phe-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N2CCC[C@@H]2C(=O)O)N)O JMBRNXUOLJFURW-BEAPCOKYSA-N 0.000 description 3
- ZESGVALRVJIVLZ-VFCFLDTKSA-N Thr-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@@H]1C(=O)O)N)O ZESGVALRVJIVLZ-VFCFLDTKSA-N 0.000 description 3
- JJNXZIPLIXIGBX-HJPIBITLSA-N Tyr-Ile-Cys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N JJNXZIPLIXIGBX-HJPIBITLSA-N 0.000 description 3
- PHKQVWWHRYUCJL-HJOGWXRNSA-N Tyr-Phe-Tyr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O PHKQVWWHRYUCJL-HJOGWXRNSA-N 0.000 description 3
- SQUMHUZLJDUROQ-YDHLFZDLSA-N Tyr-Val-Asp Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O SQUMHUZLJDUROQ-YDHLFZDLSA-N 0.000 description 3
- CFSSLXZJEMERJY-NRPADANISA-N Val-Gln-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O CFSSLXZJEMERJY-NRPADANISA-N 0.000 description 3
- HGJRMXOWUWVUOA-GVXVVHGQSA-N Val-Leu-Gln Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N HGJRMXOWUWVUOA-GVXVVHGQSA-N 0.000 description 3
- KISFXYYRKKNLOP-IHRRRGAJSA-N Val-Phe-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)O)N KISFXYYRKKNLOP-IHRRRGAJSA-N 0.000 description 3
- KRAHMIJVUPUOTQ-DCAQKATOSA-N Val-Ser-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N KRAHMIJVUPUOTQ-DCAQKATOSA-N 0.000 description 3
- CEKSLIVSNNGOKH-KZVJFYERSA-N Val-Thr-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](C(C)C)N)O CEKSLIVSNNGOKH-KZVJFYERSA-N 0.000 description 3
- ZLNYBMWGPOKSLW-LSJOCFKGSA-N Val-Val-Asp Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O ZLNYBMWGPOKSLW-LSJOCFKGSA-N 0.000 description 3
- JVGDAEKKZKKZFO-RCWTZXSCSA-N Val-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](C(C)C)N)O JVGDAEKKZKKZFO-RCWTZXSCSA-N 0.000 description 3
- IEDXPSOJFSVCKU-HOKPPMCLSA-N [4-[[(2S)-5-(carbamoylamino)-2-[[(2S)-2-[6-(2,5-dioxopyrrolidin-1-yl)hexanoylamino]-3-methylbutanoyl]amino]pentanoyl]amino]phenyl]methyl N-[(2S)-1-[[(2S)-1-[[(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]-methylamino]-3-methyl-1-oxobutan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]-N-methylcarbamate Chemical compound CC[C@H](C)[C@@H]([C@@H](CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C)[C@@H](O)c1ccccc1)OC)N(C)C(=O)[C@@H](NC(=O)[C@H](C(C)C)N(C)C(=O)OCc1ccc(NC(=O)[C@H](CCCNC(N)=O)NC(=O)[C@@H](NC(=O)CCCCCN2C(=O)CCC2=O)C(C)C)cc1)C(C)C IEDXPSOJFSVCKU-HOKPPMCLSA-N 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 108010047495 alanylglycine Proteins 0.000 description 3
- 108010009111 arginyl-glycyl-glutamic acid Proteins 0.000 description 3
- 229940049706 benzodiazepine Drugs 0.000 description 3
- 238000004364 calculation method Methods 0.000 description 3
- VSJKWCGYPAHWDS-FQEVSTJZSA-N camptothecin Chemical compound C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-FQEVSTJZSA-N 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 239000003638 chemical reducing agent Substances 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 239000012636 effector Substances 0.000 description 3
- 201000004101 esophageal cancer Diseases 0.000 description 3
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 3
- 108010013768 glutamyl-aspartyl-proline Proteins 0.000 description 3
- 108010049041 glutamylalanine Proteins 0.000 description 3
- 108010000434 glycyl-alanyl-leucine Proteins 0.000 description 3
- 108010050475 glycyl-leucyl-tyrosine Proteins 0.000 description 3
- 108010015792 glycyllysine Proteins 0.000 description 3
- 238000009169 immunotherapy Methods 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 239000003112 inhibitor Substances 0.000 description 3
- 108010057821 leucylproline Proteins 0.000 description 3
- 108010003700 lysyl aspartic acid Proteins 0.000 description 3
- 230000036961 partial effect Effects 0.000 description 3
- 108010024654 phenylalanyl-prolyl-alanine Proteins 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 102000005962 receptors Human genes 0.000 description 3
- 108020003175 receptors Proteins 0.000 description 3
- 108010048397 seryl-lysyl-leucine Proteins 0.000 description 3
- 150000003384 small molecules Chemical class 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 108010071097 threonyl-lysyl-proline Proteins 0.000 description 3
- 108010071635 tyrosyl-prolyl-arginine Proteins 0.000 description 3
- 238000000108 ultra-filtration Methods 0.000 description 3
- 108010052774 valyl-lysyl-glycyl-phenylalanyl-tyrosine Proteins 0.000 description 3
- NVKAWKQGWWIWPM-ABEVXSGRSA-N 17-β-hydroxy-5-α-Androstan-3-one Chemical compound C1C(=O)CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CC[C@H]21 NVKAWKQGWWIWPM-ABEVXSGRSA-N 0.000 description 2
- CXRCVCURMBFFOL-FXQIFTODSA-N Ala-Ala-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O CXRCVCURMBFFOL-FXQIFTODSA-N 0.000 description 2
- WCBVQNZTOKJWJS-ACZMJKKPSA-N Ala-Cys-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(O)=O WCBVQNZTOKJWJS-ACZMJKKPSA-N 0.000 description 2
- OBVSBEYOMDWLRJ-BFHQHQDPSA-N Ala-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@H](C)N OBVSBEYOMDWLRJ-BFHQHQDPSA-N 0.000 description 2
- SUHLZMHFRALVSY-YUMQZZPRSA-N Ala-Lys-Gly Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)C)C(=O)NCC(O)=O SUHLZMHFRALVSY-YUMQZZPRSA-N 0.000 description 2
- MDNAVFBZPROEHO-UHFFFAOYSA-N Ala-Lys-Val Natural products CC(C)C(C(O)=O)NC(=O)C(NC(=O)C(C)N)CCCCN MDNAVFBZPROEHO-UHFFFAOYSA-N 0.000 description 2
- ARHJJAAWNWOACN-FXQIFTODSA-N Ala-Ser-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O ARHJJAAWNWOACN-FXQIFTODSA-N 0.000 description 2
- YJHKTAMKPGFJCT-NRPADANISA-N Ala-Val-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O YJHKTAMKPGFJCT-NRPADANISA-N 0.000 description 2
- 101100330723 Arabidopsis thaliana DAR2 gene Proteins 0.000 description 2
- ZUVMUOOHJYNJPP-XIRDDKMYSA-N Arg-Trp-Gln Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(N)=O)C(O)=O ZUVMUOOHJYNJPP-XIRDDKMYSA-N 0.000 description 2
- ULBHWNVWSCJLCO-NHCYSSNCSA-N Arg-Val-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCN=C(N)N ULBHWNVWSCJLCO-NHCYSSNCSA-N 0.000 description 2
- WONGRTVAMHFGBE-WDSKDSINSA-N Asn-Gly-Gln Chemical compound C(CC(=O)N)[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC(=O)N)N WONGRTVAMHFGBE-WDSKDSINSA-N 0.000 description 2
- OOXUBGLNDRGOKT-FXQIFTODSA-N Asn-Ser-Arg Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O OOXUBGLNDRGOKT-FXQIFTODSA-N 0.000 description 2
- PUUPMDXIHCOPJU-HJGDQZAQSA-N Asn-Thr-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O PUUPMDXIHCOPJU-HJGDQZAQSA-N 0.000 description 2
- YNQIDCRRTWGHJD-ZLUOBGJFSA-N Asp-Asn-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC(O)=O YNQIDCRRTWGHJD-ZLUOBGJFSA-N 0.000 description 2
- WMLFFCRUSPNENW-ZLUOBGJFSA-N Asp-Ser-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O WMLFFCRUSPNENW-ZLUOBGJFSA-N 0.000 description 2
- VNXQRBXEQXLERQ-CIUDSAMLSA-N Asp-Ser-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(=O)O)N VNXQRBXEQXLERQ-CIUDSAMLSA-N 0.000 description 2
- 206010009944 Colon cancer Diseases 0.000 description 2
- YMBAVNPKBWHDAW-CIUDSAMLSA-N Cys-Asp-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CS)N YMBAVNPKBWHDAW-CIUDSAMLSA-N 0.000 description 2
- 102000001301 EGF receptor Human genes 0.000 description 2
- 108060006698 EGF receptor Proteins 0.000 description 2
- 208000022072 Gallbladder Neoplasms Diseases 0.000 description 2
- XOKGKOQWADCLFQ-GARJFASQSA-N Gln-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCC(=O)N)N)C(=O)O XOKGKOQWADCLFQ-GARJFASQSA-N 0.000 description 2
- NVEASDQHBRZPSU-BQBZGAKWSA-N Gln-Gln-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O NVEASDQHBRZPSU-BQBZGAKWSA-N 0.000 description 2
- XKBASPWPBXNVLQ-WDSKDSINSA-N Gln-Gly-Asn Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O XKBASPWPBXNVLQ-WDSKDSINSA-N 0.000 description 2
- FGYPOQPQTUNESW-IUCAKERBSA-N Gln-Gly-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CCC(=O)N)N FGYPOQPQTUNESW-IUCAKERBSA-N 0.000 description 2
- LHMWTCWZARHLPV-CIUDSAMLSA-N Gln-Met-Ser Chemical compound CSCC[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCC(=O)N)N LHMWTCWZARHLPV-CIUDSAMLSA-N 0.000 description 2
- NYCVMJGIJYQWDO-CIUDSAMLSA-N Gln-Ser-Arg Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NYCVMJGIJYQWDO-CIUDSAMLSA-N 0.000 description 2
- LPIKVBWNNVFHCQ-GUBZILKMSA-N Gln-Ser-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O LPIKVBWNNVFHCQ-GUBZILKMSA-N 0.000 description 2
- NHMRJKKAVMENKJ-WDCWCFNPSA-N Gln-Thr-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O NHMRJKKAVMENKJ-WDCWCFNPSA-N 0.000 description 2
- GJBUAAAIZSRCDC-GVXVVHGQSA-N Glu-Leu-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O GJBUAAAIZSRCDC-GVXVVHGQSA-N 0.000 description 2
- SUIAHERNFYRBDZ-GVXVVHGQSA-N Glu-Lys-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O SUIAHERNFYRBDZ-GVXVVHGQSA-N 0.000 description 2
- DMYACXMQUABZIQ-NRPADANISA-N Glu-Ser-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O DMYACXMQUABZIQ-NRPADANISA-N 0.000 description 2
- BIRKKBCSAIHDDF-WDSKDSINSA-N Gly-Glu-Cys Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CS)C(O)=O BIRKKBCSAIHDDF-WDSKDSINSA-N 0.000 description 2
- BUEFQXUHTUZXHR-LURJTMIESA-N Gly-Gly-Pro zwitterion Chemical compound NCC(=O)NCC(=O)N1CCC[C@H]1C(O)=O BUEFQXUHTUZXHR-LURJTMIESA-N 0.000 description 2
- GMTXWRIDLGTVFC-IUCAKERBSA-N Gly-Lys-Glu Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O GMTXWRIDLGTVFC-IUCAKERBSA-N 0.000 description 2
- GBYYQVBXFVDJPJ-WLTAIBSBSA-N Gly-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)CN)O GBYYQVBXFVDJPJ-WLTAIBSBSA-N 0.000 description 2
- FYVHHKMHFPMBBG-GUBZILKMSA-N His-Gln-Asp Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N FYVHHKMHFPMBBG-GUBZILKMSA-N 0.000 description 2
- TTYKEFZRLKQTHH-MELADBBJSA-N His-Lys-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CC2=CN=CN2)N)C(=O)O TTYKEFZRLKQTHH-MELADBBJSA-N 0.000 description 2
- HZWWOGWOBQBETJ-CUJWVEQBSA-N His-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N)O HZWWOGWOBQBETJ-CUJWVEQBSA-N 0.000 description 2
- CSTDQOOBZBAJKE-BWAGICSOSA-N His-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CC2=CN=CN2)N)O CSTDQOOBZBAJKE-BWAGICSOSA-N 0.000 description 2
- DFJJAVZIHDFOGQ-MNXVOIDGSA-N Ile-Glu-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N DFJJAVZIHDFOGQ-MNXVOIDGSA-N 0.000 description 2
- QQFSKBMCAKWHLG-UHFFFAOYSA-N Ile-Phe-Pro-Pro Chemical compound C1CCC(C(=O)N2C(CCC2)C(O)=O)N1C(=O)C(NC(=O)C(N)C(C)CC)CC1=CC=CC=C1 QQFSKBMCAKWHLG-UHFFFAOYSA-N 0.000 description 2
- VGSPNSSCMOHRRR-BJDJZHNGSA-N Ile-Ser-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O)N VGSPNSSCMOHRRR-BJDJZHNGSA-N 0.000 description 2
- RCFDOSNHHZGBOY-UHFFFAOYSA-N L-isoleucyl-L-alanine Natural products CCC(C)C(N)C(=O)NC(C)C(O)=O RCFDOSNHHZGBOY-UHFFFAOYSA-N 0.000 description 2
- WSGXUIQTEZDVHJ-GARJFASQSA-N Leu-Ala-Pro Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@@H]1C(O)=O WSGXUIQTEZDVHJ-GARJFASQSA-N 0.000 description 2
- OIARJGNVARWKFP-YUMQZZPRSA-N Leu-Asn-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O OIARJGNVARWKFP-YUMQZZPRSA-N 0.000 description 2
- BTNXKBVLWJBTNR-SRVKXCTJSA-N Leu-His-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(N)=O)C(O)=O BTNXKBVLWJBTNR-SRVKXCTJSA-N 0.000 description 2
- SBANPBVRHYIMRR-UHFFFAOYSA-N Leu-Ser-Pro Natural products CC(C)CC(N)C(=O)NC(CO)C(=O)N1CCCC1C(O)=O SBANPBVRHYIMRR-UHFFFAOYSA-N 0.000 description 2
- ILDSIMPXNFWKLH-KATARQTJSA-N Leu-Thr-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O ILDSIMPXNFWKLH-KATARQTJSA-N 0.000 description 2
- AIQWYVFNBNNOLU-RHYQMDGZSA-N Leu-Thr-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O AIQWYVFNBNNOLU-RHYQMDGZSA-N 0.000 description 2
- YQFZRHYZLARWDY-IHRRRGAJSA-N Leu-Val-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN YQFZRHYZLARWDY-IHRRRGAJSA-N 0.000 description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 2
- MWVUEPNEPWMFBD-SRVKXCTJSA-N Lys-Cys-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@H](C(O)=O)CCCCN MWVUEPNEPWMFBD-SRVKXCTJSA-N 0.000 description 2
- OIQSIMFSVLLWBX-VOAKCMCISA-N Lys-Leu-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OIQSIMFSVLLWBX-VOAKCMCISA-N 0.000 description 2
- SBQDRNOLGSYHQA-YUMQZZPRSA-N Lys-Ser-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SBQDRNOLGSYHQA-YUMQZZPRSA-N 0.000 description 2
- YKBSXQFZWFXFIB-VOAKCMCISA-N Lys-Thr-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CCCCN)C(O)=O YKBSXQFZWFXFIB-VOAKCMCISA-N 0.000 description 2
- QLFAPXUXEBAWEK-NHCYSSNCSA-N Lys-Val-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O QLFAPXUXEBAWEK-NHCYSSNCSA-N 0.000 description 2
- XABXVVSWUVCZST-GVXVVHGQSA-N Lys-Val-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCCN XABXVVSWUVCZST-GVXVVHGQSA-N 0.000 description 2
- UGCIQUYEJIEHKX-GVXVVHGQSA-N Lys-Val-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O UGCIQUYEJIEHKX-GVXVVHGQSA-N 0.000 description 2
- VOOINLQYUZOREH-SRVKXCTJSA-N Met-Gln-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCSC)N VOOINLQYUZOREH-SRVKXCTJSA-N 0.000 description 2
- RKIIYGUHIQJCBW-SRVKXCTJSA-N Met-His-Glu Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(O)=O RKIIYGUHIQJCBW-SRVKXCTJSA-N 0.000 description 2
- FWAHLGXNBLWIKB-NAKRPEOUSA-N Met-Ile-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCSC FWAHLGXNBLWIKB-NAKRPEOUSA-N 0.000 description 2
- IHRFZLQEQVHXFA-RHYQMDGZSA-N Met-Thr-Lys Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCCCN IHRFZLQEQVHXFA-RHYQMDGZSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 206010057249 Phagocytosis Diseases 0.000 description 2
- HXSUFWQYLPKEHF-IHRRRGAJSA-N Phe-Asn-Arg Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N HXSUFWQYLPKEHF-IHRRRGAJSA-N 0.000 description 2
- BIYWZVCPZIFGPY-QWRGUYRKSA-N Phe-Gly-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](CO)C(O)=O BIYWZVCPZIFGPY-QWRGUYRKSA-N 0.000 description 2
- JDMKQHSHKJHAHR-UHFFFAOYSA-N Phe-Phe-Leu-Tyr Natural products C=1C=C(O)C=CC=1CC(C(O)=O)NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)CC1=CC=CC=C1 JDMKQHSHKJHAHR-UHFFFAOYSA-N 0.000 description 2
- ZJPGOXWRFNKIQL-JYJNAYRXSA-N Phe-Pro-Pro Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(O)=O)C1=CC=CC=C1 ZJPGOXWRFNKIQL-JYJNAYRXSA-N 0.000 description 2
- UAYHMOIGIQZLFR-NHCYSSNCSA-N Pro-Gln-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O UAYHMOIGIQZLFR-NHCYSSNCSA-N 0.000 description 2
- KIPIKSXPPLABPN-CIUDSAMLSA-N Pro-Glu-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H]1CCCN1 KIPIKSXPPLABPN-CIUDSAMLSA-N 0.000 description 2
- HWLKHNDRXWTFTN-GUBZILKMSA-N Pro-Pro-Cys Chemical compound C1C[C@H](NC1)C(=O)N2CCC[C@H]2C(=O)N[C@@H](CS)C(=O)O HWLKHNDRXWTFTN-GUBZILKMSA-N 0.000 description 2
- MKGIILKDUGDRRO-FXQIFTODSA-N Pro-Ser-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H]1CCCN1 MKGIILKDUGDRRO-FXQIFTODSA-N 0.000 description 2
- KHRLUIPIMIQFGT-AVGNSLFASA-N Pro-Val-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O KHRLUIPIMIQFGT-AVGNSLFASA-N 0.000 description 2
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 2
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 2
- QPFJSHSJFIYDJZ-GHCJXIJMSA-N Ser-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CO QPFJSHSJFIYDJZ-GHCJXIJMSA-N 0.000 description 2
- XNCUYZKGQOCOQH-YUMQZZPRSA-N Ser-Leu-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O XNCUYZKGQOCOQH-YUMQZZPRSA-N 0.000 description 2
- YUJLIIRMIAGMCQ-CIUDSAMLSA-N Ser-Leu-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YUJLIIRMIAGMCQ-CIUDSAMLSA-N 0.000 description 2
- JWOBLHJRDADHLN-KKUMJFAQSA-N Ser-Leu-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JWOBLHJRDADHLN-KKUMJFAQSA-N 0.000 description 2
- RHAPJNVNWDBFQI-BQBZGAKWSA-N Ser-Pro-Gly Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O RHAPJNVNWDBFQI-BQBZGAKWSA-N 0.000 description 2
- OZPDGESCTGGNAD-CIUDSAMLSA-N Ser-Ser-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CO OZPDGESCTGGNAD-CIUDSAMLSA-N 0.000 description 2
- CUXJENOFJXOSOZ-BIIVOSGPSA-N Ser-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CO)N)C(=O)O CUXJENOFJXOSOZ-BIIVOSGPSA-N 0.000 description 2
- NADLKBTYNKUJEP-KATARQTJSA-N Ser-Thr-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O NADLKBTYNKUJEP-KATARQTJSA-N 0.000 description 2
- SIEBDTCABMZCLF-XGEHTFHBSA-N Ser-Val-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SIEBDTCABMZCLF-XGEHTFHBSA-N 0.000 description 2
- 102100038192 Serine/threonine-protein kinase TBK1 Human genes 0.000 description 2
- 102100023085 Serine/threonine-protein kinase mTOR Human genes 0.000 description 2
- 108010065917 TOR Serine-Threonine Kinases Proteins 0.000 description 2
- DWYAUVCQDTZIJI-VZFHVOOUSA-N Thr-Ala-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O DWYAUVCQDTZIJI-VZFHVOOUSA-N 0.000 description 2
- KGKWKSSSQGGYAU-SUSMZKCASA-N Thr-Gln-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N)O KGKWKSSSQGGYAU-SUSMZKCASA-N 0.000 description 2
- GKWNLDNXMMLRMC-GLLZPBPUSA-N Thr-Glu-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N)O GKWNLDNXMMLRMC-GLLZPBPUSA-N 0.000 description 2
- YOOAQCZYZHGUAZ-KATARQTJSA-N Thr-Leu-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YOOAQCZYZHGUAZ-KATARQTJSA-N 0.000 description 2
- VRUFCJZQDACGLH-UVOCVTCTSA-N Thr-Leu-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VRUFCJZQDACGLH-UVOCVTCTSA-N 0.000 description 2
- IJVNLNRVDUTWDD-MEYUZBJRSA-N Thr-Leu-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O IJVNLNRVDUTWDD-MEYUZBJRSA-N 0.000 description 2
- WRUWXBBEFUTJOU-XGEHTFHBSA-N Thr-Met-Ser Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CO)C(=O)O)N)O WRUWXBBEFUTJOU-XGEHTFHBSA-N 0.000 description 2
- XKWABWFMQXMUMT-HJGDQZAQSA-N Thr-Pro-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O XKWABWFMQXMUMT-HJGDQZAQSA-N 0.000 description 2
- XHWCDRUPDNSDAZ-XKBZYTNZSA-N Thr-Ser-Glu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N)O XHWCDRUPDNSDAZ-XKBZYTNZSA-N 0.000 description 2
- 102000002689 Toll-like receptor Human genes 0.000 description 2
- 108020000411 Toll-like receptor Proteins 0.000 description 2
- UDCHKDYNMRJYMI-QEJZJMRPSA-N Trp-Glu-Ser Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O UDCHKDYNMRJYMI-QEJZJMRPSA-N 0.000 description 2
- XGFGVFMXDXALEV-XIRDDKMYSA-N Trp-Leu-Asn Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N XGFGVFMXDXALEV-XIRDDKMYSA-N 0.000 description 2
- NLWCSMOXNKBRLC-WDSOQIARSA-N Trp-Lys-Val Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O NLWCSMOXNKBRLC-WDSOQIARSA-N 0.000 description 2
- UOXPLPBMEPLZBW-WDSOQIARSA-N Trp-Val-Lys Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(O)=O)=CNC2=C1 UOXPLPBMEPLZBW-WDSOQIARSA-N 0.000 description 2
- QYSBJAUCUKHSLU-JYJNAYRXSA-N Tyr-Arg-Val Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(O)=O QYSBJAUCUKHSLU-JYJNAYRXSA-N 0.000 description 2
- MDXLPNRXCFOBTL-BZSNNMDCSA-N Tyr-Ser-Tyr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O MDXLPNRXCFOBTL-BZSNNMDCSA-N 0.000 description 2
- PWKMJDQXKCENMF-MEYUZBJRSA-N Tyr-Thr-Leu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O PWKMJDQXKCENMF-MEYUZBJRSA-N 0.000 description 2
- CVUDMNSZAIZFAE-UHFFFAOYSA-N Val-Arg-Pro Natural products NC(N)=NCCCC(NC(=O)C(N)C(C)C)C(=O)N1CCCC1C(O)=O CVUDMNSZAIZFAE-UHFFFAOYSA-N 0.000 description 2
- OACSGBOREVRSME-NHCYSSNCSA-N Val-His-Asn Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](CC(N)=O)C(O)=O OACSGBOREVRSME-NHCYSSNCSA-N 0.000 description 2
- HJSLDXZAZGFPDK-ULQDDVLXSA-N Val-Phe-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](C(C)C)N HJSLDXZAZGFPDK-ULQDDVLXSA-N 0.000 description 2
- SSYBNWFXCFNRFN-GUBZILKMSA-N Val-Pro-Ser Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O SSYBNWFXCFNRFN-GUBZILKMSA-N 0.000 description 2
- KSFXWENSJABBFI-ZKWXMUAHSA-N Val-Ser-Asn Chemical compound [H]N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O KSFXWENSJABBFI-ZKWXMUAHSA-N 0.000 description 2
- SDHZOOIGIUEPDY-JYJNAYRXSA-N Val-Ser-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CO)NC(=O)[C@@H](N)C(C)C)C(O)=O)=CNC2=C1 SDHZOOIGIUEPDY-JYJNAYRXSA-N 0.000 description 2
- BZDGLJPROOOUOZ-XGEHTFHBSA-N Val-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](C(C)C)N)O BZDGLJPROOOUOZ-XGEHTFHBSA-N 0.000 description 2
- TVGWMCTYUFBXAP-QTKMDUPCSA-N Val-Thr-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](C(C)C)N)O TVGWMCTYUFBXAP-QTKMDUPCSA-N 0.000 description 2
- PGBMPFKFKXYROZ-UFYCRDLUSA-N Val-Tyr-Phe Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)O)N PGBMPFKFKXYROZ-UFYCRDLUSA-N 0.000 description 2
- ZHWZDZFWBXWPDW-GUBZILKMSA-N Val-Val-Cys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CS)C(O)=O ZHWZDZFWBXWPDW-GUBZILKMSA-N 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- 230000003698 anagen phase Effects 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 208000029742 colonic neoplasm Diseases 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 239000000562 conjugate Substances 0.000 description 2
- 239000013068 control sample Substances 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 238000010494 dissociation reaction Methods 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 201000010175 gallbladder cancer Diseases 0.000 description 2
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 2
- 108010048994 glycyl-tyrosyl-alanine Proteins 0.000 description 2
- 201000010536 head and neck cancer Diseases 0.000 description 2
- 208000014829 head and neck neoplasm Diseases 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 239000002955 immunomodulating agent Substances 0.000 description 2
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 2
- 108010034529 leucyl-lysine Proteins 0.000 description 2
- 108010073472 leucyl-prolyl-proline Proteins 0.000 description 2
- 108010091871 leucylmethionine Proteins 0.000 description 2
- 201000007270 liver cancer Diseases 0.000 description 2
- 208000014018 liver neoplasm Diseases 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 201000005202 lung cancer Diseases 0.000 description 2
- 208000020816 lung neoplasm Diseases 0.000 description 2
- 230000036210 malignancy Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 238000010899 nucleation Methods 0.000 description 2
- 230000008520 organization Effects 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 230000008782 phagocytosis Effects 0.000 description 2
- 230000004481 post-translational protein modification Effects 0.000 description 2
- 108010070643 prolylglutamic acid Proteins 0.000 description 2
- 108010090894 prolylleucine Proteins 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 238000001542 size-exclusion chromatography Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 210000001578 tight junction Anatomy 0.000 description 2
- 229960000575 trastuzumab Drugs 0.000 description 2
- LEAHFJQFYSDGGP-UHFFFAOYSA-K trisodium;dihydrogen phosphate;hydrogen phosphate Chemical compound [Na+].[Na+].[Na+].OP(O)([O-])=O.OP([O-])([O-])=O LEAHFJQFYSDGGP-UHFFFAOYSA-K 0.000 description 2
- 108010079202 tyrosyl-alanyl-cysteine Proteins 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- MFRNYXJJRJQHNW-DEMKXPNLSA-N (2s)-2-[[(2r,3r)-3-methoxy-3-[(2s)-1-[(3r,4s,5s)-3-methoxy-5-methyl-4-[methyl-[(2s)-3-methyl-2-[[(2s)-3-methyl-2-(methylamino)butanoyl]amino]butanoyl]amino]heptanoyl]pyrrolidin-2-yl]-2-methylpropanoyl]amino]-3-phenylpropanoic acid 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(O)=O)CC1=CC=CC=C1 MFRNYXJJRJQHNW-DEMKXPNLSA-N 0.000 description 1
- ZQJHYRVSKHGGJY-YPKJBDGSSA-N (2s,3r)-2-[[(2s)-2-[[(2s)-1-[(2s)-2-amino-3-(4-hydroxyphenyl)propanoyl]pyrrolidine-2-carbonyl]amino]-3-phenylpropanoyl]amino]-3-hydroxybutanoic acid Chemical compound C([C@@H](C(=O)N[C@@H]([C@H](O)C)C(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=CC=C1 ZQJHYRVSKHGGJY-YPKJBDGSSA-N 0.000 description 1
- DQFQCHIDRBIESA-UHFFFAOYSA-N 1-benzazepine Chemical compound N1C=CC=CC2=CC=CC=C12 DQFQCHIDRBIESA-UHFFFAOYSA-N 0.000 description 1
- NFGXHKASABOEEW-UHFFFAOYSA-N 1-methylethyl 11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate Chemical compound COC(C)(C)CCCC(C)CC=CC(C)=CC(=O)OC(C)C NFGXHKASABOEEW-UHFFFAOYSA-N 0.000 description 1
- 101150028074 2 gene Proteins 0.000 description 1
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- FJHBVJOVLFPMQE-QFIPXVFZSA-N 7-Ethyl-10-Hydroxy-Camptothecin Chemical compound C1=C(O)C=C2C(CC)=C(CN3C(C4=C([C@@](C(=O)OC4)(O)CC)C=C33)=O)C3=NC2=C1 FJHBVJOVLFPMQE-QFIPXVFZSA-N 0.000 description 1
- YYSWCHMLFJLLBJ-ZLUOBGJFSA-N Ala-Ala-Ser Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YYSWCHMLFJLLBJ-ZLUOBGJFSA-N 0.000 description 1
- JBGSZRYCXBPWGX-BQBZGAKWSA-N Ala-Arg-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](N)C)CCCN=C(N)N JBGSZRYCXBPWGX-BQBZGAKWSA-N 0.000 description 1
- KUDREHRZRIVKHS-UWJYBYFXSA-N Ala-Asp-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KUDREHRZRIVKHS-UWJYBYFXSA-N 0.000 description 1
- DPNZTBKGAUAZQU-DLOVCJGASA-N Ala-Leu-His Chemical compound C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N DPNZTBKGAUAZQU-DLOVCJGASA-N 0.000 description 1
- MEFILNJXAVSUTO-JXUBOQSCSA-N Ala-Leu-Thr Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MEFILNJXAVSUTO-JXUBOQSCSA-N 0.000 description 1
- OINVDEKBKBCPLX-JXUBOQSCSA-N Ala-Lys-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OINVDEKBKBCPLX-JXUBOQSCSA-N 0.000 description 1
- DYXOFPBJBAHWFY-JBDRJPRFSA-N Ala-Ser-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](C)N DYXOFPBJBAHWFY-JBDRJPRFSA-N 0.000 description 1
- IOFVWPYSRSCWHI-JXUBOQSCSA-N Ala-Thr-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](C)N IOFVWPYSRSCWHI-JXUBOQSCSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 101100330727 Arabidopsis thaliana DAR6 gene Proteins 0.000 description 1
- OZNSCVPYWZRQPY-CIUDSAMLSA-N Arg-Asp-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O OZNSCVPYWZRQPY-CIUDSAMLSA-N 0.000 description 1
- HPKSHFSEXICTLI-CIUDSAMLSA-N Arg-Glu-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O HPKSHFSEXICTLI-CIUDSAMLSA-N 0.000 description 1
- PNQWAUXQDBIJDY-GUBZILKMSA-N Arg-Glu-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O PNQWAUXQDBIJDY-GUBZILKMSA-N 0.000 description 1
- AOHKLEBWKMKITA-IHRRRGAJSA-N Arg-Phe-Ser Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N AOHKLEBWKMKITA-IHRRRGAJSA-N 0.000 description 1
- ZJBUILVYSXQNSW-YTWAJWBKSA-N Arg-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N)O ZJBUILVYSXQNSW-YTWAJWBKSA-N 0.000 description 1
- XRNXPIGJPQHCPC-RCWTZXSCSA-N Arg-Thr-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CCCNC(N)=N)[C@@H](C)O)C(O)=O XRNXPIGJPQHCPC-RCWTZXSCSA-N 0.000 description 1
- XYOVHPDDWCEUDY-CIUDSAMLSA-N Asn-Ala-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O XYOVHPDDWCEUDY-CIUDSAMLSA-N 0.000 description 1
- GXMSVVBIAMWMKO-BQBZGAKWSA-N Asn-Arg-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CCCN=C(N)N GXMSVVBIAMWMKO-BQBZGAKWSA-N 0.000 description 1
- QNJIRRVTOXNGMH-GUBZILKMSA-N Asn-Gln-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CC(N)=O QNJIRRVTOXNGMH-GUBZILKMSA-N 0.000 description 1
- WQLJRNRLHWJIRW-KKUMJFAQSA-N Asn-His-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CC(=O)N)N)O WQLJRNRLHWJIRW-KKUMJFAQSA-N 0.000 description 1
- RBOBTTLFPRSXKZ-BZSNNMDCSA-N Asn-Phe-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O RBOBTTLFPRSXKZ-BZSNNMDCSA-N 0.000 description 1
- SONUFGRSSMFHFN-IMJSIDKUSA-N Asn-Ser Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(O)=O SONUFGRSSMFHFN-IMJSIDKUSA-N 0.000 description 1
- JWQWPRCDYWNVNM-ACZMJKKPSA-N Asn-Ser-Gln Chemical compound C(CC(=O)N)[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(=O)N)N JWQWPRCDYWNVNM-ACZMJKKPSA-N 0.000 description 1
- HPBNLFLSSQDFQW-WHFBIAKZSA-N Asn-Ser-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O HPBNLFLSSQDFQW-WHFBIAKZSA-N 0.000 description 1
- MKJBPDLENBUHQU-CIUDSAMLSA-N Asn-Ser-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O MKJBPDLENBUHQU-CIUDSAMLSA-N 0.000 description 1
- UGXYFDQFLVCDFC-CIUDSAMLSA-N Asn-Ser-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O UGXYFDQFLVCDFC-CIUDSAMLSA-N 0.000 description 1
- VLDRQOHCMKCXLY-SRVKXCTJSA-N Asn-Ser-Phe Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O VLDRQOHCMKCXLY-SRVKXCTJSA-N 0.000 description 1
- KZYSHAMXEBPJBD-JRQIVUDYSA-N Asn-Thr-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KZYSHAMXEBPJBD-JRQIVUDYSA-N 0.000 description 1
- XYBJLTKSGFBLCS-QXEWZRGKSA-N Asp-Arg-Val Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H](C(C)C)C(O)=O)NC(=O)[C@@H](N)CC(O)=O XYBJLTKSGFBLCS-QXEWZRGKSA-N 0.000 description 1
- HSWYMWGDMPLTTH-FXQIFTODSA-N Asp-Glu-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O HSWYMWGDMPLTTH-FXQIFTODSA-N 0.000 description 1
- SNDBKTFJWVEVPO-WHFBIAKZSA-N Asp-Gly-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(O)=O SNDBKTFJWVEVPO-WHFBIAKZSA-N 0.000 description 1
- QNFRBNZGVVKBNJ-PEFMBERDSA-N Asp-Ile-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)O)N QNFRBNZGVVKBNJ-PEFMBERDSA-N 0.000 description 1
- DPNWSMBUYCLEDG-CIUDSAMLSA-N Asp-Lys-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O DPNWSMBUYCLEDG-CIUDSAMLSA-N 0.000 description 1
- DONWIPDSZZJHHK-HJGDQZAQSA-N Asp-Lys-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(=O)O)N)O DONWIPDSZZJHHK-HJGDQZAQSA-N 0.000 description 1
- CUQDCPXNZPDYFQ-ZLUOBGJFSA-N Asp-Ser-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O CUQDCPXNZPDYFQ-ZLUOBGJFSA-N 0.000 description 1
- YIDFBWRHIYOYAA-LKXGYXEUSA-N Asp-Ser-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O YIDFBWRHIYOYAA-LKXGYXEUSA-N 0.000 description 1
- XYPJXLLXNSAWHZ-SRVKXCTJSA-N Asp-Ser-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O XYPJXLLXNSAWHZ-SRVKXCTJSA-N 0.000 description 1
- CZIVKMOEXPILDK-SRVKXCTJSA-N Asp-Tyr-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(O)=O CZIVKMOEXPILDK-SRVKXCTJSA-N 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- 229940124291 BTK inhibitor Drugs 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 208000003174 Brain Neoplasms Diseases 0.000 description 1
- PLXLYXLUCNZSAA-QLXKLKPCSA-N CC[C@@]1(O)C(=O)OCC2=C1C=C1N(CC3=C1N=C1C=C(F)C(C)=C4CC[C@H](NC(=O)CO)C3=C14)C2=O Chemical compound CC[C@@]1(O)C(=O)OCC2=C1C=C1N(CC3=C1N=C1C=C(F)C(C)=C4CC[C@H](NC(=O)CO)C3=C14)C2=O PLXLYXLUCNZSAA-QLXKLKPCSA-N 0.000 description 1
- 102100040835 Claudin-18 Human genes 0.000 description 1
- VIRYODQIWJNWNU-NRPADANISA-N Cys-Glu-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CS)N VIRYODQIWJNWNU-NRPADANISA-N 0.000 description 1
- OZHXXYOHPLLLMI-CIUDSAMLSA-N Cys-Lys-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O OZHXXYOHPLLLMI-CIUDSAMLSA-N 0.000 description 1
- KSMSFCBQBQPFAD-GUBZILKMSA-N Cys-Pro-Pro Chemical compound SC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 KSMSFCBQBQPFAD-GUBZILKMSA-N 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 239000012624 DNA alkylating agent Substances 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 108090000331 Firefly luciferases Proteins 0.000 description 1
- 206010061968 Gastric neoplasm Diseases 0.000 description 1
- SOBBAYVQSNXYPQ-ACZMJKKPSA-N Gln-Asn-Asn Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O SOBBAYVQSNXYPQ-ACZMJKKPSA-N 0.000 description 1
- ZPDVKYLJTOFQJV-WDSKDSINSA-N Gln-Asn-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O ZPDVKYLJTOFQJV-WDSKDSINSA-N 0.000 description 1
- DHNWZLGBTPUTQQ-QEJZJMRPSA-N Gln-Asp-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCC(=O)N)N DHNWZLGBTPUTQQ-QEJZJMRPSA-N 0.000 description 1
- CITDWMLWXNUQKD-FXQIFTODSA-N Gln-Gln-Asn Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N CITDWMLWXNUQKD-FXQIFTODSA-N 0.000 description 1
- SMLDOQHTOAAFJQ-WDSKDSINSA-N Gln-Gly-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CO)C(O)=O SMLDOQHTOAAFJQ-WDSKDSINSA-N 0.000 description 1
- OOLCSQQPSLIETN-JYJNAYRXSA-N Gln-His-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CCC(=O)N)N)O OOLCSQQPSLIETN-JYJNAYRXSA-N 0.000 description 1
- IULKWYSYZSURJK-AVGNSLFASA-N Gln-Leu-Lys Chemical compound NC(=O)CC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O IULKWYSYZSURJK-AVGNSLFASA-N 0.000 description 1
- XUMFMAVDHQDATI-DCAQKATOSA-N Gln-Pro-Arg Chemical compound NC(=O)CC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(O)=O XUMFMAVDHQDATI-DCAQKATOSA-N 0.000 description 1
- HMIXCETWRYDVMO-GUBZILKMSA-N Gln-Pro-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O HMIXCETWRYDVMO-GUBZILKMSA-N 0.000 description 1
- UKKNTTCNGZLJEX-WHFBIAKZSA-N Gln-Ser Chemical compound NC(=O)CC[C@H](N)C(=O)N[C@@H](CO)C(O)=O UKKNTTCNGZLJEX-WHFBIAKZSA-N 0.000 description 1
- OKARHJKJTKFQBM-ACZMJKKPSA-N Gln-Ser-Asn Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)N)C(=O)O)N OKARHJKJTKFQBM-ACZMJKKPSA-N 0.000 description 1
- OSCLNNWLKKIQJM-WDSKDSINSA-N Gln-Ser-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)NCC(O)=O OSCLNNWLKKIQJM-WDSKDSINSA-N 0.000 description 1
- VLOLPWWCNKWRNB-LOKLDPHHSA-N Gln-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O VLOLPWWCNKWRNB-LOKLDPHHSA-N 0.000 description 1
- QZQYITIKPAUDGN-GVXVVHGQSA-N Gln-Val-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CCC(=O)N)N QZQYITIKPAUDGN-GVXVVHGQSA-N 0.000 description 1
- JJKKWYQVHRUSDG-GUBZILKMSA-N Glu-Ala-Lys Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O JJKKWYQVHRUSDG-GUBZILKMSA-N 0.000 description 1
- WOMUDRVDJMHTCV-DCAQKATOSA-N Glu-Arg-Arg Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O WOMUDRVDJMHTCV-DCAQKATOSA-N 0.000 description 1
- PAQUJCSYVIBPLC-AVGNSLFASA-N Glu-Asp-Phe Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 PAQUJCSYVIBPLC-AVGNSLFASA-N 0.000 description 1
- CLROYXHHUZELFX-FXQIFTODSA-N Glu-Gln-Asp Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O CLROYXHHUZELFX-FXQIFTODSA-N 0.000 description 1
- PVBBEKPHARMPHX-DCAQKATOSA-N Glu-Gln-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CCC(O)=O PVBBEKPHARMPHX-DCAQKATOSA-N 0.000 description 1
- IVGJYOOGJLFKQE-AVGNSLFASA-N Glu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N IVGJYOOGJLFKQE-AVGNSLFASA-N 0.000 description 1
- YGLCLCMAYUYZSG-AVGNSLFASA-N Glu-Lys-His Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 YGLCLCMAYUYZSG-AVGNSLFASA-N 0.000 description 1
- QDMVXRNLOPTPIE-WDCWCFNPSA-N Glu-Lys-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QDMVXRNLOPTPIE-WDCWCFNPSA-N 0.000 description 1
- AAJHGGDRKHYSDH-GUBZILKMSA-N Glu-Pro-Gln Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CCC(=O)O)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O AAJHGGDRKHYSDH-GUBZILKMSA-N 0.000 description 1
- JWNZHMSRZXXGTM-XKBZYTNZSA-N Glu-Ser-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O JWNZHMSRZXXGTM-XKBZYTNZSA-N 0.000 description 1
- DLISPGXMKZTWQG-IFFSRLJSSA-N Glu-Thr-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O DLISPGXMKZTWQG-IFFSRLJSSA-N 0.000 description 1
- BPCLDCNZBUYGOD-BPUTZDHNSA-N Glu-Trp-Glu Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)N)C(=O)N[C@@H](CCC(O)=O)C(O)=O)=CNC2=C1 BPCLDCNZBUYGOD-BPUTZDHNSA-N 0.000 description 1
- NTNUEBVGKMVANB-NHCYSSNCSA-N Glu-Val-Met Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCSC)C(O)=O NTNUEBVGKMVANB-NHCYSSNCSA-N 0.000 description 1
- WGYHAAXZWPEBDQ-IFFSRLJSSA-N Glu-Val-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O WGYHAAXZWPEBDQ-IFFSRLJSSA-N 0.000 description 1
- UXJHNZODTMHWRD-WHFBIAKZSA-N Gly-Asn-Ala Chemical compound [H]NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(O)=O UXJHNZODTMHWRD-WHFBIAKZSA-N 0.000 description 1
- BGVYNAQWHSTTSP-BYULHYEWSA-N Gly-Asn-Ile Chemical compound [H]NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BGVYNAQWHSTTSP-BYULHYEWSA-N 0.000 description 1
- XCLCVBYNGXEVDU-WHFBIAKZSA-N Gly-Asn-Ser Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O XCLCVBYNGXEVDU-WHFBIAKZSA-N 0.000 description 1
- GRIRDMVMJJDZKV-RCOVLWMOSA-N Gly-Asn-Val Chemical compound [H]NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O GRIRDMVMJJDZKV-RCOVLWMOSA-N 0.000 description 1
- NNCSJUBVFBDDLC-YUMQZZPRSA-N Gly-Leu-Ser Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O NNCSJUBVFBDDLC-YUMQZZPRSA-N 0.000 description 1
- ICUTTWWCDIIIEE-BQBZGAKWSA-N Gly-Met-Asn Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)CN ICUTTWWCDIIIEE-BQBZGAKWSA-N 0.000 description 1
- HAOUOFNNJJLVNS-BQBZGAKWSA-N Gly-Pro-Ser Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O HAOUOFNNJJLVNS-BQBZGAKWSA-N 0.000 description 1
- JSLVAHYTAJJEQH-QWRGUYRKSA-N Gly-Ser-Phe Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 JSLVAHYTAJJEQH-QWRGUYRKSA-N 0.000 description 1
- CQMFNTVQVLQRLT-JHEQGTHGSA-N Gly-Thr-Gln Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(O)=O CQMFNTVQVLQRLT-JHEQGTHGSA-N 0.000 description 1
- XHVONGZZVUUORG-WEDXCCLWSA-N Gly-Thr-Lys Chemical compound NCC(=O)N[C@@H]([C@H](O)C)C(=O)N[C@H](C(O)=O)CCCCN XHVONGZZVUUORG-WEDXCCLWSA-N 0.000 description 1
- WTUSRDZLLWGYAT-KCTSRDHCSA-N Gly-Trp-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)CN WTUSRDZLLWGYAT-KCTSRDHCSA-N 0.000 description 1
- 208000017891 HER2 positive breast carcinoma Diseases 0.000 description 1
- MAABHGXCIBEYQR-XVYDVKMFSA-N His-Asn-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CN=CN1)N MAABHGXCIBEYQR-XVYDVKMFSA-N 0.000 description 1
- HIAHVKLTHNOENC-HGNGGELXSA-N His-Glu-Ala Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O HIAHVKLTHNOENC-HGNGGELXSA-N 0.000 description 1
- TVMNTHXFRSXZGR-IHRRRGAJSA-N His-Lys-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O TVMNTHXFRSXZGR-IHRRRGAJSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000749329 Homo sapiens Claudin-18 Proteins 0.000 description 1
- 101000665442 Homo sapiens Serine/threonine-protein kinase TBK1 Proteins 0.000 description 1
- 101000653510 Homo sapiens TATA box-binding protein-like 2 Proteins 0.000 description 1
- MKWSZEHGHSLNPF-NAKRPEOUSA-N Ile-Ala-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)O)N MKWSZEHGHSLNPF-NAKRPEOUSA-N 0.000 description 1
- NZGTYCMLUGYMCV-XUXIUFHCSA-N Ile-Lys-Arg Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N NZGTYCMLUGYMCV-XUXIUFHCSA-N 0.000 description 1
- JHNJNTMTZHEDLJ-NAKRPEOUSA-N Ile-Ser-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O JHNJNTMTZHEDLJ-NAKRPEOUSA-N 0.000 description 1
- FBGXMKUWQFPHFB-JBDRJPRFSA-N Ile-Ser-Cys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O)N FBGXMKUWQFPHFB-JBDRJPRFSA-N 0.000 description 1
- ANTFEOSJMAUGIB-KNZXXDILSA-N Ile-Thr-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@@H]1C(=O)O)N ANTFEOSJMAUGIB-KNZXXDILSA-N 0.000 description 1
- DGTOKVBDZXJHNZ-WZLNRYEVSA-N Ile-Thr-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N DGTOKVBDZXJHNZ-WZLNRYEVSA-N 0.000 description 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 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
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- 239000005411 L01XE02 - Gefitinib Substances 0.000 description 1
- 239000005551 L01XE03 - Erlotinib Substances 0.000 description 1
- 239000002147 L01XE04 - Sunitinib Substances 0.000 description 1
- 239000002118 L01XE12 - Vandetanib Substances 0.000 description 1
- 239000002177 L01XE27 - Ibrutinib Substances 0.000 description 1
- DBVWMYGBVFCRBE-CIUDSAMLSA-N Leu-Asn-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O DBVWMYGBVFCRBE-CIUDSAMLSA-N 0.000 description 1
- OGCQGUIWMSBHRZ-CIUDSAMLSA-N Leu-Asn-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O OGCQGUIWMSBHRZ-CIUDSAMLSA-N 0.000 description 1
- PVMPDMIKUVNOBD-CIUDSAMLSA-N Leu-Asp-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O PVMPDMIKUVNOBD-CIUDSAMLSA-N 0.000 description 1
- QLQHWWCSCLZUMA-KKUMJFAQSA-N Leu-Asp-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 QLQHWWCSCLZUMA-KKUMJFAQSA-N 0.000 description 1
- RSFGIMMPWAXNML-MNXVOIDGSA-N Leu-Gln-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O RSFGIMMPWAXNML-MNXVOIDGSA-N 0.000 description 1
- CQGSYZCULZMEDE-UHFFFAOYSA-N Leu-Gln-Pro Natural products CC(C)CC(N)C(=O)NC(CCC(N)=O)C(=O)N1CCCC1C(O)=O CQGSYZCULZMEDE-UHFFFAOYSA-N 0.000 description 1
- GPICTNQYKHHHTH-GUBZILKMSA-N Leu-Gln-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O GPICTNQYKHHHTH-GUBZILKMSA-N 0.000 description 1
- HPBCTWSUJOGJSH-MNXVOIDGSA-N Leu-Glu-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HPBCTWSUJOGJSH-MNXVOIDGSA-N 0.000 description 1
- VCHVSKNMTXWIIP-SRVKXCTJSA-N Leu-Lys-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O VCHVSKNMTXWIIP-SRVKXCTJSA-N 0.000 description 1
- FIICHHJDINDXKG-IHPCNDPISA-N Leu-Lys-Trp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O FIICHHJDINDXKG-IHPCNDPISA-N 0.000 description 1
- UHNQRAFSEBGZFZ-YESZJQIVSA-N Leu-Phe-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N2CCC[C@@H]2C(=O)O)N UHNQRAFSEBGZFZ-YESZJQIVSA-N 0.000 description 1
- DPURXCQCHSQPAN-AVGNSLFASA-N Leu-Pro-Pro Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 DPURXCQCHSQPAN-AVGNSLFASA-N 0.000 description 1
- RGUXWMDNCPMQFB-YUMQZZPRSA-N Leu-Ser-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RGUXWMDNCPMQFB-YUMQZZPRSA-N 0.000 description 1
- AMSSKPUHBUQBOQ-SRVKXCTJSA-N Leu-Ser-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O)N AMSSKPUHBUQBOQ-SRVKXCTJSA-N 0.000 description 1
- LINKCQUOMUDLKN-KATARQTJSA-N Leu-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC(C)C)N)O LINKCQUOMUDLKN-KATARQTJSA-N 0.000 description 1
- DAYQSYGBCUKVKT-VOAKCMCISA-N Leu-Thr-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(O)=O DAYQSYGBCUKVKT-VOAKCMCISA-N 0.000 description 1
- FBNPMTNBFFAMMH-UHFFFAOYSA-N Leu-Val-Arg Natural products CC(C)CC(N)C(=O)NC(C(C)C)C(=O)NC(C(O)=O)CCCN=C(N)N FBNPMTNBFFAMMH-UHFFFAOYSA-N 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- WSXTWLJHTLRFLW-SRVKXCTJSA-N Lys-Ala-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O WSXTWLJHTLRFLW-SRVKXCTJSA-N 0.000 description 1
- IXHKPDJKKCUKHS-GARJFASQSA-N Lys-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCCN)N IXHKPDJKKCUKHS-GARJFASQSA-N 0.000 description 1
- SWWCDAGDQHTKIE-RHYQMDGZSA-N Lys-Arg-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SWWCDAGDQHTKIE-RHYQMDGZSA-N 0.000 description 1
- LMVOVCYVZBBWQB-SRVKXCTJSA-N Lys-Asp-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN LMVOVCYVZBBWQB-SRVKXCTJSA-N 0.000 description 1
- ODUQLUADRKMHOZ-JYJNAYRXSA-N Lys-Glu-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CCCCN)N)O ODUQLUADRKMHOZ-JYJNAYRXSA-N 0.000 description 1
- GQZMPWBZQALKJO-UWVGGRQHSA-N Lys-Gly-Arg Chemical compound [H]N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O GQZMPWBZQALKJO-UWVGGRQHSA-N 0.000 description 1
- FGMHXLULNHTPID-KKUMJFAQSA-N Lys-His-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CCCCN)C(O)=O)CC1=CN=CN1 FGMHXLULNHTPID-KKUMJFAQSA-N 0.000 description 1
- AIRZWUMAHCDDHR-KKUMJFAQSA-N Lys-Leu-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O AIRZWUMAHCDDHR-KKUMJFAQSA-N 0.000 description 1
- YTJFXEDRUOQGSP-DCAQKATOSA-N Lys-Pro-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O YTJFXEDRUOQGSP-DCAQKATOSA-N 0.000 description 1
- SQXZLVXQXWILKW-KKUMJFAQSA-N Lys-Ser-Phe Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O SQXZLVXQXWILKW-KKUMJFAQSA-N 0.000 description 1
- PSVAVKGDUAKZKU-BZSNNMDCSA-N Lys-Tyr-His Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)NC(=O)[C@H](CCCCN)N)O PSVAVKGDUAKZKU-BZSNNMDCSA-N 0.000 description 1
- GILLQRYAWOMHED-DCAQKATOSA-N Lys-Val-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCCN GILLQRYAWOMHED-DCAQKATOSA-N 0.000 description 1
- HMZPYMSEAALNAE-ULQDDVLXSA-N Lys-Val-Tyr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O HMZPYMSEAALNAE-ULQDDVLXSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- PNDCUTDWYVKBHX-IHRRRGAJSA-N Met-Asp-Tyr Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 PNDCUTDWYVKBHX-IHRRRGAJSA-N 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 206010059282 Metastases to central nervous system Diseases 0.000 description 1
- 102000029749 Microtubule Human genes 0.000 description 1
- 108091022875 Microtubule Proteins 0.000 description 1
- 108010057466 NF-kappa B Proteins 0.000 description 1
- 102000003945 NF-kappa B Human genes 0.000 description 1
- 238000011789 NOD SCID mouse Methods 0.000 description 1
- 102000003940 Occludin Human genes 0.000 description 1
- 108090000304 Occludin Proteins 0.000 description 1
- 239000012661 PARP inhibitor Substances 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 239000012807 PCR reagent Substances 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- LBSARGIQACMGDF-WBAXXEDZSA-N Phe-Ala-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 LBSARGIQACMGDF-WBAXXEDZSA-N 0.000 description 1
- BWTKUQPNOMMKMA-FIRPJDEBSA-N Phe-Ile-Phe Chemical compound C([C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 BWTKUQPNOMMKMA-FIRPJDEBSA-N 0.000 description 1
- JHSRGEODDALISP-XVSYOHENSA-N Phe-Thr-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O JHSRGEODDALISP-XVSYOHENSA-N 0.000 description 1
- GNRMAQSIROFNMI-IXOXFDKPSA-N Phe-Thr-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O GNRMAQSIROFNMI-IXOXFDKPSA-N 0.000 description 1
- KIQUCMUULDXTAZ-HJOGWXRNSA-N Phe-Tyr-Tyr Chemical compound N[C@@H](Cc1ccccc1)C(=O)N[C@@H](Cc1ccc(O)cc1)C(=O)N[C@@H](Cc1ccc(O)cc1)C(O)=O KIQUCMUULDXTAZ-HJOGWXRNSA-N 0.000 description 1
- 229940122907 Phosphatase inhibitor Drugs 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 229940121906 Poly ADP ribose polymerase inhibitor Drugs 0.000 description 1
- CLNJSLSHKJECME-BQBZGAKWSA-N Pro-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H]1CCCN1 CLNJSLSHKJECME-BQBZGAKWSA-N 0.000 description 1
- UIMCLYYSUCIUJM-UWVGGRQHSA-N Pro-Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 UIMCLYYSUCIUJM-UWVGGRQHSA-N 0.000 description 1
- FMLRRBDLBJLJIK-DCAQKATOSA-N Pro-Leu-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CCCN1 FMLRRBDLBJLJIK-DCAQKATOSA-N 0.000 description 1
- GMJDSFYVTAMIBF-FXQIFTODSA-N Pro-Ser-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O GMJDSFYVTAMIBF-FXQIFTODSA-N 0.000 description 1
- PRKWBYCXBBSLSK-GUBZILKMSA-N Pro-Ser-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O PRKWBYCXBBSLSK-GUBZILKMSA-N 0.000 description 1
- VVAWNPIOYXAMAL-KJEVXHAQSA-N Pro-Thr-Tyr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O VVAWNPIOYXAMAL-KJEVXHAQSA-N 0.000 description 1
- 101710096655 Probable acetoacetate decarboxylase 1 Proteins 0.000 description 1
- 101710096660 Probable acetoacetate decarboxylase 2 Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 1
- 102000001708 Protein Isoforms Human genes 0.000 description 1
- 108010029485 Protein Isoforms Proteins 0.000 description 1
- 108091027981 Response element Proteins 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- YUSRGTQIPCJNHQ-CIUDSAMLSA-N Ser-Arg-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O YUSRGTQIPCJNHQ-CIUDSAMLSA-N 0.000 description 1
- HZWAHWQZPSXNCB-BPUTZDHNSA-N Ser-Arg-Trp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O HZWAHWQZPSXNCB-BPUTZDHNSA-N 0.000 description 1
- VAUMZJHYZQXZBQ-WHFBIAKZSA-N Ser-Asn-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O VAUMZJHYZQXZBQ-WHFBIAKZSA-N 0.000 description 1
- UGJRQLURDVGULT-LKXGYXEUSA-N Ser-Asn-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O UGJRQLURDVGULT-LKXGYXEUSA-N 0.000 description 1
- FTVRVZNYIYWJGB-ACZMJKKPSA-N Ser-Asp-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O FTVRVZNYIYWJGB-ACZMJKKPSA-N 0.000 description 1
- BNFVPSRLHHPQKS-WHFBIAKZSA-N Ser-Asp-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O BNFVPSRLHHPQKS-WHFBIAKZSA-N 0.000 description 1
- ULVMNZOKDBHKKI-ACZMJKKPSA-N Ser-Gln-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O ULVMNZOKDBHKKI-ACZMJKKPSA-N 0.000 description 1
- ZOHGLPQGEHSLPD-FXQIFTODSA-N Ser-Gln-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O ZOHGLPQGEHSLPD-FXQIFTODSA-N 0.000 description 1
- HJEBZBMOTCQYDN-ACZMJKKPSA-N Ser-Glu-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O HJEBZBMOTCQYDN-ACZMJKKPSA-N 0.000 description 1
- BPMRXBZYPGYPJN-WHFBIAKZSA-N Ser-Gly-Asn Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O BPMRXBZYPGYPJN-WHFBIAKZSA-N 0.000 description 1
- SFTZWNJFZYOLBD-ZDLURKLDSA-N Ser-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CO SFTZWNJFZYOLBD-ZDLURKLDSA-N 0.000 description 1
- HBTCFCHYALPXME-HTFCKZLJSA-N Ser-Ile-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HBTCFCHYALPXME-HTFCKZLJSA-N 0.000 description 1
- ZOPISOXXPQNOCO-SVSWQMSJSA-N Ser-Ile-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](CO)N ZOPISOXXPQNOCO-SVSWQMSJSA-N 0.000 description 1
- NLOAIFSWUUFQFR-CIUDSAMLSA-N Ser-Leu-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O NLOAIFSWUUFQFR-CIUDSAMLSA-N 0.000 description 1
- GJFYFGOEWLDQGW-GUBZILKMSA-N Ser-Leu-Gln Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CO)N GJFYFGOEWLDQGW-GUBZILKMSA-N 0.000 description 1
- UBRMZSHOOIVJPW-SRVKXCTJSA-N Ser-Leu-Lys Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O UBRMZSHOOIVJPW-SRVKXCTJSA-N 0.000 description 1
- KCGIREHVWRXNDH-GARJFASQSA-N Ser-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N KCGIREHVWRXNDH-GARJFASQSA-N 0.000 description 1
- PPNPDKGQRFSCAC-CIUDSAMLSA-N Ser-Lys-Asp Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)CO)C(=O)N[C@@H](CC(O)=O)C(O)=O PPNPDKGQRFSCAC-CIUDSAMLSA-N 0.000 description 1
- RRVFEDGUXSYWOW-BZSNNMDCSA-N Ser-Phe-Phe Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O RRVFEDGUXSYWOW-BZSNNMDCSA-N 0.000 description 1
- CKDXFSPMIDSMGV-GUBZILKMSA-N Ser-Pro-Val Chemical compound [H]N[C@@H](CO)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O CKDXFSPMIDSMGV-GUBZILKMSA-N 0.000 description 1
- ZKOKTQPHFMRSJP-YJRXYDGGSA-N Ser-Thr-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZKOKTQPHFMRSJP-YJRXYDGGSA-N 0.000 description 1
- AXKJPUBALUNJEO-UBHSHLNASA-N Ser-Trp-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(N)=O)C(O)=O AXKJPUBALUNJEO-UBHSHLNASA-N 0.000 description 1
- FGBLCMLXHRPVOF-IHRRRGAJSA-N Ser-Tyr-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FGBLCMLXHRPVOF-IHRRRGAJSA-N 0.000 description 1
- BIWBTRRBHIEVAH-IHPCNDPISA-N Ser-Tyr-Trp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O BIWBTRRBHIEVAH-IHPCNDPISA-N 0.000 description 1
- YEDSOSIKVUMIJE-DCAQKATOSA-N Ser-Val-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O YEDSOSIKVUMIJE-DCAQKATOSA-N 0.000 description 1
- 101710106944 Serine/threonine-protein kinase TBK1 Proteins 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 102100030631 TATA box-binding protein-like 2 Human genes 0.000 description 1
- PXQUBKWZENPDGE-CIQUZCHMSA-N Thr-Ala-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C)NC(=O)[C@H]([C@@H](C)O)N PXQUBKWZENPDGE-CIQUZCHMSA-N 0.000 description 1
- UKBSDLHIKIXJKH-HJGDQZAQSA-N Thr-Arg-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O UKBSDLHIKIXJKH-HJGDQZAQSA-N 0.000 description 1
- CEXFELBFVHLYDZ-XGEHTFHBSA-N Thr-Arg-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O CEXFELBFVHLYDZ-XGEHTFHBSA-N 0.000 description 1
- JVTHIXKSVYEWNI-JRQIVUDYSA-N Thr-Asn-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JVTHIXKSVYEWNI-JRQIVUDYSA-N 0.000 description 1
- YOSLMIPKOUAHKI-OLHMAJIHSA-N Thr-Asp-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O YOSLMIPKOUAHKI-OLHMAJIHSA-N 0.000 description 1
- AQAMPXBRJJWPNI-JHEQGTHGSA-N Thr-Gly-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O AQAMPXBRJJWPNI-JHEQGTHGSA-N 0.000 description 1
- CYVQBKQYQGEELV-NKIYYHGXSA-N Thr-His-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N)O CYVQBKQYQGEELV-NKIYYHGXSA-N 0.000 description 1
- SXAGUVRFGJSFKC-ZEILLAHLSA-N Thr-His-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SXAGUVRFGJSFKC-ZEILLAHLSA-N 0.000 description 1
- XSEPSRUDSPHMPX-KATARQTJSA-N Thr-Lys-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O XSEPSRUDSPHMPX-KATARQTJSA-N 0.000 description 1
- OHDXOXIZXSFCDN-RCWTZXSCSA-N Thr-Met-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O OHDXOXIZXSFCDN-RCWTZXSCSA-N 0.000 description 1
- IEZVHOULSUULHD-XGEHTFHBSA-N Thr-Ser-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O IEZVHOULSUULHD-XGEHTFHBSA-N 0.000 description 1
- CYCGARJWIQWPQM-YJRXYDGGSA-N Thr-Tyr-Ser Chemical compound C[C@@H](O)[C@H]([NH3+])C(=O)N[C@H](C(=O)N[C@@H](CO)C([O-])=O)CC1=CC=C(O)C=C1 CYCGARJWIQWPQM-YJRXYDGGSA-N 0.000 description 1
- OGOYMQWIWHGTGH-KZVJFYERSA-N Thr-Val-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O OGOYMQWIWHGTGH-KZVJFYERSA-N 0.000 description 1
- FYBFTPLPAXZBOY-KKHAAJSZSA-N Thr-Val-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O FYBFTPLPAXZBOY-KKHAAJSZSA-N 0.000 description 1
- BKVICMPZWRNWOC-RHYQMDGZSA-N Thr-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)[C@@H](C)O BKVICMPZWRNWOC-RHYQMDGZSA-N 0.000 description 1
- AVYVKJMBNLPWRX-WFBYXXMGSA-N Trp-Ala-Ser Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O)=CNC2=C1 AVYVKJMBNLPWRX-WFBYXXMGSA-N 0.000 description 1
- SCQBNMKLZVCXNX-ZFWWWQNUSA-N Trp-Arg-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)NCC(=O)O)N SCQBNMKLZVCXNX-ZFWWWQNUSA-N 0.000 description 1
- AZBIIKDSDLVJAK-VHWLVUOQSA-N Trp-Ile-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N AZBIIKDSDLVJAK-VHWLVUOQSA-N 0.000 description 1
- 108090000704 Tubulin Proteins 0.000 description 1
- 102000004243 Tubulin Human genes 0.000 description 1
- CDRYEAWHKJSGAF-BPNCWPANSA-N Tyr-Ala-Met Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(=O)N[C@@H](CCSC)C(O)=O CDRYEAWHKJSGAF-BPNCWPANSA-N 0.000 description 1
- ARPONUQDNWLXOZ-KKUMJFAQSA-N Tyr-Gln-Arg Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O ARPONUQDNWLXOZ-KKUMJFAQSA-N 0.000 description 1
- SLCSPPCQWUHPPO-JYJNAYRXSA-N Tyr-Glu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 SLCSPPCQWUHPPO-JYJNAYRXSA-N 0.000 description 1
- BSCBBPKDVOZICB-KKUMJFAQSA-N Tyr-Leu-Asp Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O BSCBBPKDVOZICB-KKUMJFAQSA-N 0.000 description 1
- GZUIDWDVMWZSMI-KKUMJFAQSA-N Tyr-Lys-Cys Chemical compound NCCCC[C@@H](C(=O)N[C@@H](CS)C(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 GZUIDWDVMWZSMI-KKUMJFAQSA-N 0.000 description 1
- LMKKMCGTDANZTR-BZSNNMDCSA-N Tyr-Phe-Asp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C1=CC=C(O)C=C1 LMKKMCGTDANZTR-BZSNNMDCSA-N 0.000 description 1
- AUZADXNWQMBZOO-JYJNAYRXSA-N Tyr-Pro-Arg Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O)C1=CC=C(O)C=C1 AUZADXNWQMBZOO-JYJNAYRXSA-N 0.000 description 1
- YYLHVUCSTXXKBS-IHRRRGAJSA-N Tyr-Pro-Ser Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O YYLHVUCSTXXKBS-IHRRRGAJSA-N 0.000 description 1
- VYQQQIRHIFALGE-UWJYBYFXSA-N Tyr-Ser-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 VYQQQIRHIFALGE-UWJYBYFXSA-N 0.000 description 1
- MQGGXGKQSVEQHR-KKUMJFAQSA-N Tyr-Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 MQGGXGKQSVEQHR-KKUMJFAQSA-N 0.000 description 1
- NHOVZGFNTGMYMI-KKUMJFAQSA-N Tyr-Ser-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 NHOVZGFNTGMYMI-KKUMJFAQSA-N 0.000 description 1
- ZZDYJFVIKVSUFA-WLTAIBSBSA-N Tyr-Thr-Gly Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O ZZDYJFVIKVSUFA-WLTAIBSBSA-N 0.000 description 1
- YOTRXXBHTZHKLU-BVSLBCMMSA-N Tyr-Trp-Met Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(O)=O)C1=CC=C(O)C=C1 YOTRXXBHTZHKLU-BVSLBCMMSA-N 0.000 description 1
- VJOWWOGRNXRQMF-UVBJJODRSA-N Val-Ala-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](C)NC(=O)[C@@H](N)C(C)C)C(O)=O)=CNC2=C1 VJOWWOGRNXRQMF-UVBJJODRSA-N 0.000 description 1
- LHADRQBREKTRLR-DCAQKATOSA-N Val-Cys-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](C(C)C)N LHADRQBREKTRLR-DCAQKATOSA-N 0.000 description 1
- OXVPMZVGCAPFIG-BQFCYCMXSA-N Val-Gln-Trp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N OXVPMZVGCAPFIG-BQFCYCMXSA-N 0.000 description 1
- KZKMBGXCNLPYKD-YEPSODPASA-N Val-Gly-Thr Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O KZKMBGXCNLPYKD-YEPSODPASA-N 0.000 description 1
- XTDDIVQWDXMRJL-IHRRRGAJSA-N Val-Leu-His Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](C(C)C)N XTDDIVQWDXMRJL-IHRRRGAJSA-N 0.000 description 1
- SYSWVVCYSXBVJG-RHYQMDGZSA-N Val-Leu-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C(C)C)N)O SYSWVVCYSXBVJG-RHYQMDGZSA-N 0.000 description 1
- VCIYTVOBLZHFSC-XHSDSOJGSA-N Val-Phe-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N2CCC[C@@H]2C(=O)O)N VCIYTVOBLZHFSC-XHSDSOJGSA-N 0.000 description 1
- ZXYPHBKIZLAQTL-QXEWZRGKSA-N Val-Pro-Asp Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(=O)O)C(=O)O)N ZXYPHBKIZLAQTL-QXEWZRGKSA-N 0.000 description 1
- PGQUDQYHWICSAB-NAKRPEOUSA-N Val-Ser-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](C(C)C)N PGQUDQYHWICSAB-NAKRPEOUSA-N 0.000 description 1
- NZYNRRGJJVSSTJ-GUBZILKMSA-N Val-Ser-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O NZYNRRGJJVSSTJ-GUBZILKMSA-N 0.000 description 1
- GVNLOVJNNDZUHS-RHYQMDGZSA-N Val-Thr-Lys Chemical compound [H]N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(O)=O GVNLOVJNNDZUHS-RHYQMDGZSA-N 0.000 description 1
- QHSSPPHOHJSTML-HOCLYGCPSA-N Val-Trp-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)NCC(=O)O)N QHSSPPHOHJSTML-HOCLYGCPSA-N 0.000 description 1
- QPJSIBAOZBVELU-BPNCWPANSA-N Val-Tyr-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](C(C)C)N QPJSIBAOZBVELU-BPNCWPANSA-N 0.000 description 1
- BGTDGENDNWGMDQ-KJEVXHAQSA-N Val-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](C(C)C)N)O BGTDGENDNWGMDQ-KJEVXHAQSA-N 0.000 description 1
- LLJLBRRXKZTTRD-GUBZILKMSA-N Val-Val-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)O)N LLJLBRRXKZTTRD-GUBZILKMSA-N 0.000 description 1
- 108091022873 acetoacetate decarboxylase Proteins 0.000 description 1
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 108010005233 alanylglutamic acid Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 108010050025 alpha-glutamyltryptophan Proteins 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000009830 antibody antigen interaction Effects 0.000 description 1
- 210000000612 antigen-presenting cell Anatomy 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 108010062796 arginyllysine Proteins 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 108010038633 aspartylglutamate Proteins 0.000 description 1
- 229960002707 bendamustine Drugs 0.000 description 1
- YTKUWDBFDASYHO-UHFFFAOYSA-N bendamustine Chemical compound ClCCN(CCCl)C1=CC=C2N(C)C(CCCC(O)=O)=NC2=C1 YTKUWDBFDASYHO-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 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 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- SQQXRXKYTKFFSM-UHFFFAOYSA-N chembl1992147 Chemical compound OC1=C(OC)C(OC)=CC=C1C1=C(C)C(C(O)=O)=NC(C=2N=C3C4=NC(C)(C)N=C4C(OC)=C(O)C3=CC=2)=C1N SQQXRXKYTKFFSM-UHFFFAOYSA-N 0.000 description 1
- 239000013626 chemical specie Substances 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 229940044683 chemotherapy drug Drugs 0.000 description 1
- 238000009104 chemotherapy regimen Methods 0.000 description 1
- 238000013375 chromatographic separation Methods 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 238000002648 combination therapy Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000011254 conventional chemotherapy Methods 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 108010081447 cytochrophin-4 Proteins 0.000 description 1
- 230000002380 cytological effect Effects 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000006240 deamidation Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 208000018459 dissociative disease Diseases 0.000 description 1
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 229940121647 egfr inhibitor Drugs 0.000 description 1
- 230000001159 endocytotic effect Effects 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 229960001433 erlotinib Drugs 0.000 description 1
- AAKJLRGGTJKAMG-UHFFFAOYSA-N erlotinib Chemical compound C=12C=C(OCCOC)C(OCCOC)=CC2=NC=NC=1NC1=CC=CC(C#C)=C1 AAKJLRGGTJKAMG-UHFFFAOYSA-N 0.000 description 1
- ZVYVPGLRVWUPMP-FYSMJZIKSA-N exatecan Chemical compound C1C[C@H](N)C2=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC3=CC(F)=C(C)C1=C32 ZVYVPGLRVWUPMP-FYSMJZIKSA-N 0.000 description 1
- 229950009429 exatecan Drugs 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 210000001156 gastric mucosa Anatomy 0.000 description 1
- 229960002584 gefitinib Drugs 0.000 description 1
- XGALLCVXEZPNRQ-UHFFFAOYSA-N gefitinib Chemical compound C=12C=C(OCCCN3CCOCC3)C(OC)=CC2=NC=NC=1NC1=CC=C(F)C(Cl)=C1 XGALLCVXEZPNRQ-UHFFFAOYSA-N 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 108010077435 glycyl-phenylalanyl-glycine Proteins 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 108010087823 glycyltyrosine Proteins 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 108010036413 histidylglycine Proteins 0.000 description 1
- 108010025306 histidylleucine Proteins 0.000 description 1
- 239000003276 histone deacetylase inhibitor Substances 0.000 description 1
- 235000003642 hunger Nutrition 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 229960001507 ibrutinib Drugs 0.000 description 1
- XYFPWWZEPKGCCK-GOSISDBHSA-N ibrutinib Chemical compound C1=2C(N)=NC=NC=2N([C@H]2CN(CCC2)C(=O)C=C)N=C1C(C=C1)=CC=C1OC1=CC=CC=C1 XYFPWWZEPKGCCK-GOSISDBHSA-N 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 238000010829 isocratic elution Methods 0.000 description 1
- 238000006317 isomerization reaction Methods 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 238000003670 luciferase enzyme activity assay Methods 0.000 description 1
- 208000037841 lung tumor Diseases 0.000 description 1
- 210000003712 lysosome Anatomy 0.000 description 1
- 230000001868 lysosomic effect Effects 0.000 description 1
- 108010064235 lysylglycine Proteins 0.000 description 1
- 108010038320 lysylphenylalanine Proteins 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 208000037819 metastatic cancer Diseases 0.000 description 1
- 208000011575 metastatic malignant neoplasm Diseases 0.000 description 1
- 210000004688 microtubule Anatomy 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- AMKBTTRWLGVRER-OFVILXPXSA-N n-[(2s)-1-[[(2s)-5-(carbamoylamino)-1-[4-(hydroxymethyl)anilino]-1-oxopentan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]-6-(2,5-dioxopyrrol-1-yl)hexanamide Chemical compound N([C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=O)C(=O)NC=1C=CC(CO)=CC=1)C(=O)CCCCCN1C(=O)C=CC1=O AMKBTTRWLGVRER-OFVILXPXSA-N 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- FAQDUNYVKQKNLD-UHFFFAOYSA-N olaparib Chemical compound FC1=CC=C(CC2=C3[CH]C=CC=C3C(=O)N=N2)C=C1C(=O)N(CC1)CCN1C(=O)C1CC1 FAQDUNYVKQKNLD-UHFFFAOYSA-N 0.000 description 1
- 229960000572 olaparib Drugs 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 238000009522 phase III clinical trial Methods 0.000 description 1
- 108010073101 phenylalanylleucine Proteins 0.000 description 1
- 108010073025 phenylalanylphenylalanine Proteins 0.000 description 1
- 108010051242 phenylalanylserine Proteins 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920006389 polyphenyl polymer Polymers 0.000 description 1
- 238000010837 poor prognosis Methods 0.000 description 1
- 238000000159 protein binding assay Methods 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 229960004641 rituximab Drugs 0.000 description 1
- 238000005464 sample preparation method Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 210000004989 spleen cell Anatomy 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 229960001796 sunitinib Drugs 0.000 description 1
- WINHZLLDWRZWRT-ATVHPVEESA-N sunitinib Chemical compound CCN(CC)CCNC(=O)C1=C(C)NC(\C=C/2C3=CC(F)=CC=C3NC\2=O)=C1C WINHZLLDWRZWRT-ATVHPVEESA-N 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000725 suspension Substances 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
- 229940126585 therapeutic drug Drugs 0.000 description 1
- 229960003989 tocilizumab Drugs 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 108010080629 tryptophan-leucine Proteins 0.000 description 1
- 230000005760 tumorsuppression Effects 0.000 description 1
- 108010051110 tyrosyl-lysine Proteins 0.000 description 1
- 108010003137 tyrosyltyrosine Proteins 0.000 description 1
- 229960000241 vandetanib Drugs 0.000 description 1
- UHTHHESEBZOYNR-UHFFFAOYSA-N vandetanib Chemical compound COC1=CC(C(/N=CN2)=N/C=3C(=CC(Br)=CC=3)F)=C2C=C1OCC1CCN(C)CC1 UHTHHESEBZOYNR-UHFFFAOYSA-N 0.000 description 1
- WAEXFXRVDQXREF-UHFFFAOYSA-N vorinostat Chemical compound ONC(=O)CCCCCCC(=O)NC1=CC=CC=C1 WAEXFXRVDQXREF-UHFFFAOYSA-N 0.000 description 1
- 229960000237 vorinostat Drugs 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Images
Classifications
-
- 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
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
-
- 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
- C07K2317/56—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
- C07K2317/50—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
- C07K2317/565—Complementarity determining region [CDR]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
- C07K2317/732—Antibody-dependent cellular cytotoxicity [ADCC]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
- C07K2317/734—Complement-dependent cytotoxicity [CDC]
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Immunology (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Organic Chemistry (AREA)
- Epidemiology (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Genetics & Genomics (AREA)
- General Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Cell Biology (AREA)
- Peptides Or Proteins (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
本发明公开了包含抗CLDN18.2的抗体或其片段的抗体药物偶联物。此外,本发明还公开了包含该抗体药物偶联物的药物组合物,以及该抗体药物偶联物和该药物组合物在制备药物中的用途。
Description
相关申请的交叉引用
本专利申请要求于2021年2月9日提交的申请号为CN202110181327.3的中国发明专利申请的优先权权益,在此将其全部内容引入作为参考。
技术领域
本发明属于生物技术领域,具体地,本发明涉及靶向CLDN18.2的抗体药物偶联物及其应用。
背景技术
近年来全球恶性肿瘤发病率逐年升高,据世界卫生组织(World HealthOrganization)和美国癌症协会(American Cancer Society)统计数据显示,0-74岁患癌症风险已上升至20.2%,其中男性患癌症风险高达22.4%,女性患癌症风险达18.2%。
胃癌是全球第五大最常见癌症和第三大癌症死亡原因。据统计,2018年新增胃癌100多万病例,死亡78.3万例。目前临床上胃癌的治疗以手术、化疗和放疗为主,尽管已有多种治疗方案,但5年生存率仍然很低。
随着靶向治疗手段的出现,针对不同途径的不同分子被开发出来用于胃癌的治疗。但是,目前研制的靶向药物仍存在不足。例如曲妥珠单抗,在Her2阳性乳腺癌中取得了成功,但是对胃癌的治疗效果并不理想;同样,针对表皮生长因子受体(epidermal growthfactor receptor,EGFR)和哺乳动物类雷帕霉素靶蛋白(mammalian target ofrapamycin,mTOR)的抑制剂在胃肿瘤的III期临床试验中也被证实无效。仍需要寻求新靶点来进行胃癌的靶向治疗。
闭合蛋白(CLDN)为一个蛋白质家族,于1998年首次由Shorichiro Tsukita等人发现,该家族的成员具有重要的机体生理作用,如参与细胞旁通透性、电导调节,维持上皮和内皮细胞间分子交换的紧密连接等。CLDN18是CLDN家族包含的至少24个成员之一,具有CLDN18.1和CLDN18.2两种异构体,其中CLDN18.2是一种胃特异性亚型,通常埋藏在胃粘膜的分化上皮细胞中,正常组织中很少或不表达,而在癌细胞中广泛表达,是一种高度选择性的分子。当恶性肿瘤发生时,细胞间分子交换的紧密连接遭到破坏,肿瘤细胞表面的CLDN18.2表位由此暴露出来,使得CLDN18.2可能成为靶向治疗的特异性位点。
已发现CLDN18.2在原发性胃癌及其转移癌中表达,在胃癌患者中,53%的患者具有CLDN18.2的高丰度表达、20%患者具有中丰度表达;而在胃癌脑转移患者的组织切片中,在转移灶中发现45.16%的患者具有CLDN18.2的高丰度表达。事实上,研究表明CLDN18.2在人类多种癌症中出现异位激活,高度选择性地、稳定地表达于特定肿瘤组织,比如在胰腺癌、食管癌、卵巢癌和肺肿瘤中。正是CLDN18.2在多个癌组织中的高度选择性表达,以及在特定癌种如胃癌、胰腺癌等患者中的高阳性率,使得CLDN18.2成为一个颇具潜力的靶向治疗与肿瘤免疫治疗新靶点,引起了国内外研究者的关注。
日本制药企业安斯泰来针对这一靶点开发了抗体药物Claudiximab(IMAB362),其在临床I期/II期显示了较好的治疗效果,2018年7月已经启动了该药物的三期临床试验。目前临床上主推的方案为胃癌EOX+Claudiximab的组合疗法,而Claudiximab的单药使用效果结果不佳。
从目前的治疗情况来看,胃癌、尤其是晚期胃癌恶性程度高、预后差,常规化疗失败后,治疗手段有限;而免疫治疗仅能使部分患者获益,而且可用的免疫治疗抑制剂数量也极为有限,目前患者能够选择的治疗药物并不多,并且客观缓解率不高。因此,仍需要开发治疗效果更为显著、适用性更广的药物,以满足临床的迫切需求。
发明内容
本发明要解决的技术问题是,通过杂交瘤筛选和人源化技术,获得特异性结合人CLDN18.2的高亲和力抗体,其中通过人源化设计,使该抗体最大程度减少鼠源氨基酸的数目,以拥有更好的体内安全性和应用前景;在此基础上,进一步筛选具有更强细胞内吞效应的抗体,将其与小分子化学药物制备成抗体药物偶联物(Antibody-Drug Conjugate,ADC),利用该抗体对CLDN18.2表达细胞的靶向性和经由靶点的强的内吞效应,结合该小分子化学药物的作用,来增强针对CLDN18.2的抗体药物的治疗效果。
因此,本发明的一个目的是提供特异性结合人CLDN18.2的抗体或其片段;其中,所述的抗体的片段涵盖抗体的各种功能性片段,例如其抗原结合部分,如Fab、F(ab’)2或scFv片段。本发明的另一个目的是提供采用该抗体或其片段制备的抗体药物偶联物。
本发明的技术方案如下。
一方面,本发明提供一种抗体药物偶联物,其包含与药物共价连接的抗CLDN18.2的抗体或其片段。
在本发明提供的抗体药物偶联物中,所述抗CLDN18.2的抗体或其片段包含重链和轻链,所述重链和轻链分别包含如下所示的重链互补决定区1至3(CDR-H1、CDR-H2和CDR-H3)和轻链互补决定区1至3(CDR-L1、CDR-L2和CDR-L3):
(i)氨基酸序列分别如SEQ ID NO:23、SEQ ID NO:26和SEQ ID NO:25所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:32、SEQ ID NO:30和SEQ ID NO:33所示的CDR-L1、CDR-L2和CDR-L3;
(ii)氨基酸序列分别如SEQ ID NO:27、SEQ ID NO:28和SEQ ID NO:25所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:32、SEQ ID NO:30和SEQ ID NO:33所示的CDR-L1、CDR-L2和CDR-L3;
(iii)氨基酸序列分别如SEQ ID NO:34、SEQ ID NO:35和SEQ ID NO:36所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:41、SEQ ID NO:30和SEQ IDNO:40所示的CDR-L1、CDR-L2和CDR-L3;
(iv)氨基酸序列分别如SEQ ID NO:37、SEQ ID NO:38和SEQ ID NO:36所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:41、SEQ ID NO:30和SEQ ID NO:43所示的CDR-L1、CDR-L2和CDR-L3;或
(v)氨基酸序列分别如SEQ ID NO:34、SEQ ID NO:35和SEQ ID NO:36所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:41、SEQ ID NO:30和SEQ ID NO:43所示的CDR-L1、CDR-L2和CDR-L3。
优选地,在本发明提供的抗体药物偶联物中,所述抗CLDN18.2的抗体或其片段的重链包含重链可变区(VH),所述重链可变区(VH)包含SEQ ID NO:7或SEQ ID NO:8所示的氨基酸序列或其变体;和/或,所述抗CLDN18.2的抗体或其片段的轻链包含轻链可变区(VL),所述轻链可变区(VL)包含:选自SEQ ID NO:11或SEQ ID NO:12所示的氨基酸序列或其变体。
或者,优选地,在本发明提供的抗体药物偶联物中,所述抗CLDN18.2的抗体或其片段的重链包含重链可变区(VH),所述重链可变区(VH)包含SEQ ID NO:15或SEQ ID NO:16所示的氨基酸序列或其变体;和/或,所述抗CLDN18.2的抗体或其片段的轻链包含轻链可变区(VL),所述轻链可变区(VL)包含选自SEQ ID NO:19或SEQ ID NO:22所示的氨基酸序列或其变体。
更优选地,在本发明提供的抗体药物偶联物中,所述抗CLDN18.2的抗体或其片段的重链和轻链分别包含如下所示的重链可变区(VH)和轻链可变区(VL):
(i)如SEQ ID NO:7所示的氨基酸序列或其变体;和,如SEQ ID NO:11所示的氨基酸序列或其变体;
(ii)如SEQ ID NO:8所示的氨基酸序列或其变体;和,如SEQ ID NO:12所示的氨基酸序列或其变体;
(iii)如SEQ ID NO:15所示的氨基酸序列或其变体;和,如SEQ ID NO:19所示的氨基酸序列或其变体;
(iv)如SEQ ID NO:16所示的氨基酸序列或其变体;和,如SEQ ID NO:22所示的氨基酸序列或其变体。
本发明的上下文中,“氨基酸序列的变体”是指与所述氨基酸序列具有至少75%序列同一性(例如至少80%、优选至少85%、更优选至少90%、进一步优选至少91%、92%、93%、94%、95%、96%、97%、98%或甚至99%同一性等≥75%的任何百分比的同一性)的氨基酸序列。
在本发明提供的抗体药物偶联物中,所述抗CLDN18.2的抗体或其片段可以为针对CLDN18.2的单克隆抗体、单链抗体、双功能抗体、单域抗体、纳米抗体、完全或部分人源化的抗体或者嵌合抗体等任意形式,或者,所述抗体或其片段为针对CLDN18.2的半抗体或半抗体的抗原结合片段,例如scFv、BsFv、dsFv、(dsFv)2、Fab、Fab'、F(ab')2或Fv;关于本发明提供的抗体的片段,优选地,所述片段为抗体的能够特异性结合CLDN18.2的任何片段。并且,就抗CLDN18.2的抗体或其片段而言,优选地,所述CLDN18.2为人CLDN18.2。
特别地,在本发明提供的抗体药物偶联物中,所述抗CLDN18.2的抗体或其片段至少包含抗体的重链可变区和轻链可变区,而这两者均可分别包括上述CDR1至CDR13以及间隔的框架区FR1至FR4,各个区域的排列方式按照FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4的顺序。在这一方面,上文所述的氨基酸序列的变体与氨基酸序列之间的“至少75%序列同一性”可能由重链可变区和轻链可变区中框架区FR1至FR4中任一个存在的差异导致,或者,也可能由重链可变区和轻链可变区以外的任意结构域或序列中存在的差异导致。这种差异可以是任何位置的氨基酸缺失、添加或置换,其中置换可以是保守置换或非保守置换。
在本发明提供的抗体药物偶联物中,优选地,所述抗CLDN18.2的抗体或其片段还包含人或鼠的恒定区,优选包含人或鼠的轻链恒定区(CL)和/或重链恒定区(CH);更优选地,所述抗体或其片段包含选自IgG、IgA、IgM、IgD或IgE的重链恒定区和/或κ或λ型轻链恒定区。
优选地,所述抗体为单克隆抗体,优选为鼠源、嵌合或人源化的单克隆抗体;更优选地,所述单克隆抗体的重链恒定区为IgG1或IgG4亚型,轻链恒定区为κ型。
根据本发明的具体实施方式,所述单克隆抗体的重链恒定区包含如SEQ ID NO:48所示的氨基酸序列或其变体。或者,根据本发明的具体实施方式,所述单克隆抗体的轻链恒定区包含如SEQ ID NO:49所示氨基酸序列或其变体。
具体地,本发明提供的抗体药物偶联物具有分子式Ab-[L-D]n,其中Ab表示本发明提供的抗CLDN18.2的抗体或其片段,L表示接头,D表示药物,n表示相对于每一分子Ab的药物平均连接数(DAR)。
在本发明提供的抗体药物偶联物中,所述药物D为细胞毒性类小分子药物和/或免疫治疗剂类小分子药物。其中,所述细胞毒性类小分子药物可以为微管蛋白抑制剂和/或拓扑异构酶I抑制剂,微管蛋白抑制剂可以为澳瑞他汀类化合物,如MMAE、MMAF等;拓扑异构酶I抑制剂可以为喜树碱类化合物,如SN38、Exatecan以及Dxd。这些化合物的结构示例见下:
所述免疫治疗剂类小分子药物可以为STING激动剂和/或TLR7/8激动剂。其中,STING激动剂可以为环二核苷酸类化合物,例如如下所示的化合物10和11等;TLR7/8激动剂可以为咪唑并喹啉类化合物和/或苯并氮卓类化合物,例如如下所示的化合物1、2、3、4、5、6、7、8和9等。
在本发明提供的抗体药物偶联物中,所述接头L可以为半胱氨酸偶联接头或赖氨酸偶联接头,其释放机制为可切割或不可切割的。可切割的接头如可选自:MC-Val-Cit-PAB、MC-Val-Ala和MC-Gly-Gly-Phe-Gly等;不可切割的接头如可选自MCC、PEG4Mal和mc等。
具体地,在本发明提供的抗体药物偶联物中,细胞毒性类药物接头(即“[L-D]”部分,在本发明的上下文中“药物接头”、“[L-D]”和“含药接头”可互换使用)可以为:
1)喜树碱类拓扑异构酶I抑制剂-ADC接头,MC-GGFG-Dxd(简称为EX)和CL2A-SN38(简称为EX2):
2)澳瑞他汀类微管蛋白抑制剂-ADC接头,MC-VC-MMAE(简称为M):
免疫治疗剂类药物接头(即“[L-D]”部分)可以为:
1)苯并氮卓类TLR8激动剂-ADC接头可以为
2)环二核苷酸类STING激动剂-ADC接头可以为:
根据本发明的具体实施方式,在本发明提供的抗体药物偶联物中的“[L-D]”为实施例部分采用的特定化合物,其中MC-GGFG-Dxd缩写为“EX”、CL2A-SN38缩写为“EX2”、MC-VC-MMAE缩写为“M”、L-A缩写为“TLR8”和L-B缩写为“STING”。相应地,本发明提供的抗体药物偶联物可命名为“抗体简称-[L-D]缩写”。
优选地,在本发明提供的抗体药物偶联物中,n为1至12、优选1至8、更优选3至8,例如4至8、5至8、6至8、乃至7至8。
另一方面,本发明还提供所述抗体药物偶联物的制备方法,所述方法包括以下步骤:
(1)用抗体与还原试剂在缓冲液中反应,得到经还原后的抗体;
(2)用药物接头(连接子-药物缀合物)与步骤(1)得到的经还原后的抗体在缓冲液与有机溶剂的混合液中进行交联,得到抗体药物偶联物。
优选地,所述方法包括以下步骤:
1)还原:将抗体原液用反应缓冲液稀释,加入抗体1.5-2.5倍摩尔比的还原剂如TCEP,反应液于室温搅拌1-4小时;
2)偶联:用反应缓冲液调整抗体还原液的浓度,再加入抗体4.0-6.0倍摩尔比的药物接头的有机溶剂如二甲基亚砜溶液,将该溶液在室温条件下继续搅拌30-60分钟;
3)纯化:将抗体偶联液浓缩,色谱分离收集主成分,超滤。
根据本发明的具体实施方式,本发明提供DAR为4的抗体药物偶联物的制备方法,包括:
1)还原:将抗体原液用反应缓冲液稀释至约10mg/mL,加入抗体1.5-2.5倍摩尔比的三(2-羧乙基)膦盐酸盐(TCEP),反应液于25℃搅动2小时;
2)偶联:将抗体还原液用反应缓冲液调整浓度至5mg/mL,再加入抗体4.0-6.0倍摩尔比的药物接头的二甲基亚砜溶液,将该溶液在室温条件下继续搅拌30-60分钟;
3)纯化:将抗体偶联液浓缩至15mg/mL左右,添加缓冲液调整电导率为100ms/cm,上样至NAP-25色谱柱,分离并收集主成分;最后超滤离心管将最终样品置换至pH值7.4的50mM的磷酸氢二钠-磷酸二氢钠缓冲盐中,除菌过滤。
根据本发明的具体实施方式,本发明提供DAR为8的抗体药物偶联物的制备方法,包括:
1)还原:将抗体原液用反应缓冲液稀释至约10mg/mL,加入抗体9.0-11倍摩尔比的三(2-羧乙基)膦盐酸盐(TCEP),反应液于25℃搅动2小时;
2)偶联:将抗体还原液用反应缓冲液调整浓度至5mg/mL,再加入抗体10-12倍摩尔比的药物接头的二甲基亚砜溶液,将该溶液在室温条件下继续搅拌30-60分钟;
3)纯化:将抗体偶联液浓缩至15mg/mL左右,添加缓冲液调整电导率为100ms/cm。上样至NAP-25色谱柱,分离并收集主成分;最后超滤离心管将最终样品置换至pH值7.4的50mM的磷酸氢二钠-磷酸二氢钠缓冲盐中,除菌过滤。
在本发明的另一方面,本发明提供一种抗CLDN18.2的抗体或其片段。上文就抗体药物偶联物中包含的抗CLDN18.2的抗体或其片段进行的描述与定义均适用于这一方面的抗CLDN18.2的抗体或其片段。
关于具体序列,优选地,本发明提供的抗CLDN18.2的抗体或其片段包含重链和轻链,所述重链和轻链分别包含如下所示的重链互补决定区1至3(CDR-H1、CDR-H2和CDR-H3)和轻链互补决定区1至3(CDR-L1、CDR-L2和CDR-L3):
(i)氨基酸序列分别如SEQ ID NO:23、SEQ ID NO:26和SEQ ID NO:25所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:32、SEQ ID NO:30和SEQ ID NO:33所示的CDR-L1、CDR-L2和CDR-L3;
(ii)氨基酸序列分别如SEQ ID NO:27、SEQ ID NO:28和SEQ ID NO:25所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:32、SEQ ID NO:30和SEQ ID NO:33所示的CDR-L1、CDR-L2和CDR-L3;
(iii)氨基酸序列分别如SEQ ID NO:34、SEQ ID NO:35和SEQ ID NO:36所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:41、SEQ ID NO:30和SEQ IDNO:40所示的CDR-L1、CDR-L2和CDR-L3;
(iv)氨基酸序列分别如SEQ ID NO:37、SEQ ID NO:38和SEQ ID NO:36所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:41、SEQ ID NO:30和SEQ ID NO:43所示的CDR-L1、CDR-L2和CDR-L3;或
(v)氨基酸序列分别如SEQ ID NO:34、SEQ ID NO:35和SEQ ID NO:36所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:41、SEQ ID NO:30和SEQ ID NO:43所示的CDR-L1、CDR-L2和CDR-L3。
优选地,所述抗CLDN18.2的抗体或其片段的重链包含重链可变区(VH),轻链包含轻链可变区(VL),所述重链可变区(VH)和轻链可变区(VL)分别包含SEQ ID NO:8所示的氨基酸序列或其变体和SEQ ID NO:12所示的氨基酸序列或其变体;或者,所述重链可变区(VH)和轻链可变区(VL)分别包含SEQ ID NO:16所示的氨基酸序列或其变体和SEQ ID NO:22所示的氨基酸序列或其变体。
又一方面,本发明还提供一种药物组合物,所述药物组合物包含本发明的抗体药物偶联物或者包含本发明的抗CLDN18.2的抗体或其片段,以及可选的药学上可接受的辅料。其中,所述辅料可以是载体或赋形剂等。
进一步地,本发明提供的药物组合物还可包含能够与本发明的抗体药物偶联物或者抗CLDN18.2的抗体或其片段联合使用的其他药物,例如化疗药物EGFR抑制剂(Gefitinib、Erlotinib、Vandetanib、Sunitinib等)、PARP抑制剂(Olaparib等)、DNA烷化剂(Bendamustine等)、HDAC抑制剂(Vorinostat等)、BTK抑制剂(Ibrutinib等)、C-met抑制剂(Tepotinib等)、紫杉醇等,或者本发明提供的药物组合物还可与其他药物或其他治疗方法如化疗方案联合使用。
又一方面,本发明提供了一种药物组合物。根据本发明的实施例,该药物组合物包含抗体药物偶联物混合物和药学上可接受的载体,所述抗体药物偶联物混合物包含多种如前述的抗体药物偶联物。
任选地,所述抗体药物偶联物混合物的平均n值为2至8。
任选地,所述抗体药物偶联物混合物包含多种所述抗体药物偶联物,每种所述抗体药物偶联物的n值为1至12、优选1至8、更优选3至8,例如4至8、5至8、6至8、乃至7至8。
再一方面,本发明还提供所述抗体药物偶联物、抗CLDN18.2的抗体或其片段或包含它们任意的药物组合物在制备药物中的用途,所述药物用于治疗与CLDN18.2相关的疾病,优选为肿瘤,更优选为癌症。进一步优选地,所述癌症选自胰腺癌、胃癌、结肠癌、食管癌、肝癌、卵巢癌、肺癌、胆囊癌和头颈癌。
又一方面,本发明还提供一种治疗疾病的方法,所述方法包括向有此需要的受试者施用本发明提供的抗体药物偶联物、抗CLDN18.2的抗体或其片段或包含它们任意的药物组合物。其中所述受试者为哺乳类动物,优选地为人。所述疾病为与CLDN18.2相关的疾病,优选为肿瘤,更优选为癌症。进一步优选地,所述癌症选自胰腺癌、胃癌、结肠癌、食管癌、肝癌、卵巢癌、肺癌、胆囊癌和头颈癌。
相对于现有技术,本发明实现了以下有益的技术效果:
首先,在本发明中使用细胞表面稳定表达人CLDN18.2的高CHOK1细胞免疫小鼠,获得分泌抗人CLDN18.2抗体的杂交瘤细胞,然后由杂交瘤细胞分泌的鼠抗构建嵌合抗体,在证明该嵌合抗体保留了鼠抗与抗原的特异性结合,进行重链和轻链可变区的人源化改造,得到了多个针对人CLDN18.2的不同人源化抗体。实验证明,本发明提供的抗CLDN18.2抗体对抗原CLDN18.2具有强亲和力,对靶点表达细胞具有显著的补体依赖性细胞毒(CDC)活性和抗体依赖性细胞毒(ADCC)活性,且活性强于同靶点的IMAB362或与其相当。特别是,与本身作为嵌合抗体的IMAB362相比,本发明提供的抗CLDN18.2抗体最大程度地减少了鼠源氨基酸,由此能够降低在人体内引起排斥免疫反应的可能性。
进一步地,实验证明,相比于IMAB362,本发明的抗CLDN18.2抗体具有更优的内吞活性,与小分子药物制备得到的抗体药物偶联物稳定性也更强。采用本发明的抗CLDN18.2抗体制备得到的ADC,在通过该抗体的靶向作用和/或Fc效应介导免疫细胞吞噬后进入细胞内部,能够有效实现所偶联的药物释放,在示例性的细胞毒性类小分子药物或免疫治疗剂类小分子药物的情况下,实现了细胞毒性类小分子药物对CLDN18.2表达肿瘤细胞的显著的杀伤作用,或实现免疫治疗剂类小分子药物对细胞内对应受体以及下游信号通路的显著的活化作用。因此,本发明提供的抗CLDN18.2抗体更适合与药物等制备成抗体药物偶联物,兼顾安全性和有效性。
附图说明
以下,结合附图来详细说明本发明的实施方案,其中:
图1:本发明的抗体与Isotype抗体的表位研究结果;
图2:本发明的抗体与细胞表面CLDN18.2的结合活性;
图3:本发明的抗体结合细胞表面CLDN18.2的内化活性;
图4:抗体药物偶联物8k13-hz35-TLR8-1103的HIC分析结果;
图5:抗体药物偶联物8k13-hz35-TLR8-1103的SEC分析结果;
图6:抗体药物偶联物8k13-hz35-STING-D4-1203的RPLC分析结果;
图7:抗体药物偶联物8k13-hz35-STING-D4-1203的SEC分析结果;
图8:抗体药物偶联物8k13-hz35-M-20200318-1的HIC分析结果;
图9:抗体药物偶联物8k13-hz35-M-20200318-1的SEC分析结果;
图10:抗体药物偶联物8k13-hz35-EX-DAR4-20200922的RPLC分析结果;
图11:抗体药物偶联物8k13-hz35-EX-DAR4-20200922的SEC分析结果;
图12:抗体药物偶联物8k13-hz35-EX-DAR8-20200421的HIC分析结果;
图13:抗体药物偶联物8k13-hz35-EX-DAR8-20200421的SEC分析结果;
图14:抗体药物偶联物8k13-hz35-EX2-DAR4-20200922的RPLC分析结果;
图15:抗体药物偶联物8k13-hz35-EX2-DAR4-20200922的SEC分析结果;
图16:抗体药物偶联物18k13-hz35-EX2-DAR8-20200922的RPLC分析结果;
图17:抗体药物偶联物18k13-hz35-EX2-DAR8-20200922的SEC分析结果;
图18:抗体药物偶联物16k15-hz35-M-20200318-1的HIC分析结果;
图19:抗体药物偶联物16k15-hz35-M-20200318-1的SEC分析结果;
图20:抗体药物偶联物16k15-hz35-EX-20200421的HIC分析结果;
图21:抗体药物偶联物16k15-hz35-EX-20200421的SEC分析结果;
图22:抗体药物偶联物IMAB362-M-1的HIC分析结果;
图23:抗体药物偶联物IMAB362-M-1的SEC分析结果;
图24:抗体药物偶联物IMAB362-M-2的HIC分析结果;
图25:抗体药物偶联物IMAB362-M-2的SEC分析结果;
图26:本发明的抗体药物偶联物对无CLDN18.2表达的细胞的杀伤效果;
图27:本发明的抗体药物偶联物对CLDN18.2高表达的细胞的杀伤效果;
图28:本发明的抗体药物偶联物对PBMC分泌促炎因子TNFα的刺激作用;
图29:本发明的抗体药物偶联物对pIRF3表达的刺激作用;
图30:本发明的抗体药物偶联物对小鼠皮下移植瘤的疗效;
图31:本发明的抗体药物偶联物对荷瘤小鼠体重的影响;
图32:本发明的抗体药物偶联物对荷瘤小鼠摄食的影响;
图33:本发明的抗体药物偶联物对小鼠皮下移植瘤的疗效;
图34:本发明的抗体药物偶联物对人胃癌KATO III/18.2小鼠皮下移植瘤的疗效(个体肿瘤重量);**P<0.001,***P<0.001,vs溶剂。
具体实施方式
以下参照具体的实施例来说明本发明。本领域技术人员能够理解,这些实施例仅用于说明本发明,其不以任何方式限制本发明的范围。
下述实施例中的实验方法,如无特殊说明,均为常规方法。下述实施例中所用的原料、试剂等,如无特殊说明,均为市售购买产品。
(一)用于制备抗体药物偶联物的抗体的制备与表征
以下实施例中:
抗原人CLDN18.2参见NP_001002026.1,抗原人CLDN18.1参见NP_057453.1。
IMAB362的重链和轻链序列参见SEQ ID NO:44和SEQ ID NO:45。
Sacituzumab的重链和轻链序列参见SEQ ID NO:46和SEQ ID NO:47。
人单克隆抗体IgG1亚类的重链恒定区序列和κ亚类的轻链恒定区序列分别示于SEQ ID NO:48和SEQ ID NO:49。
实施例1 鼠源单克隆抗体的筛选
使用细胞表面稳定表达人CLDN18.2蛋白的CHOK1细胞免疫Balb/c小鼠,1个月后用流式细胞仪术(FACS)分析小鼠血清,取血清抗体滴度高的小鼠取其脾脏,标准方法分离得到的脾细胞与骨髓瘤细胞P3X63Ag8.653使用PEG或者电融合方法进行融合。将融合后的杂交瘤细胞接种于384孔板中,培养10-14天后,取上清用FACS分析杂交瘤细胞分泌的抗体,筛选得到若干克隆,所述克隆能够结合细胞表面稳定表达人CLDN18.2蛋白的CHOK1细胞而不能够结合细胞表面稳定表达人CLDN18.1蛋白的CHOK1细胞。通过有限稀释的方法将筛选到的克隆单细胞化,3轮之后得到的每个杂交瘤细胞克隆只分泌一个抗体。
将分泌抗人CLDN18.2抗体的杂交瘤细胞扩大培养后,按照RNAfast200试剂盒(上海飞捷生物技术有限公司)说明书步骤提取细胞总RNA;利用5×PrimeScript RT MasterMix(Takara)将杂交瘤细胞总RNA反转录成cDNA;使用简并引物(Anke Krebber.1997)和Extaq PCR试剂(Takara)扩增抗体轻链可变区IgVL(κ)和重链可变区VH序列;利用PCRclean-up Gel extraction试剂盒(Macherey-Nagel公司)纯化PCR扩增产物;按照pClone007 Simple Vector Kit试剂盒(擎科生物科技有限公司)说明书将扩增PCR产物连接至T载体并转化大肠杆菌感受态细胞,菌株扩增、抽提质粒后进行DNA测序获得单克隆抗体可变区序列。见表1。
表1.鼠抗的轻重链可变区
鼠抗 | 重链可变区(VH) | 轻链可变区(VL) |
8k13 | SEQ ID NO:1 | SEQ ID NO:2 |
16k15 | SEQ ID NO:3 | SEQ ID NO:4 |
鼠抗8k13
重链可变区
>8K13_vh(SEQ ID NO:1;CDR按顺序为:SEQ ID NO:23、SEQ ID NO:24、SEQ ID NO:25)
QVHLQQSGAELVRPGSSVKISCKASGYAFSNYWMNWVRQRPGQGLEWIGQIYPGNGDTKYSGKFNSKDTLTADKSSNTAYMQLNSLTSEDSAVYFCARFYYGNVMDYWGQGTSVTVSS
轻链可变区
>8K13_vl(SEQ ID NO:2;CDR按顺序为:SEQ ID NO:29、SEQ ID NO:30、SEQ ID NO:31)
DIVLTQSPSSLTVTAGEKVTMSCKSSQTLLNGGNQKNYLTWYQQKSGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNGYSYPLTFGVGTKLELK
鼠抗16k15
重链可变区
>16K15_vh(SEQ ID NO:3;CDR按顺序为:SEQ ID NO:34、SEQ ID NO:35、SEQ IDNO:36)
EVMLVESGGGLVRPGGSLKLSCAGSGITLSTYAMSWVRQTPERRLEWVASIISGGITYYLDSVKGRFTISRDNARNILYLQMSSLRSEDTAIYYCARKYHGNALDYWGQGTSVTVSL
轻链可变区
>16K15_vl(SEQ ID NO:4;CDR按顺序为:SEQ ID NO:39、SEQ ID NO:30、SEQ IDNO:40)
DIVMTQSPSSLPVTAGETVTMRCKSSQSLLNSGNQRNYLTWYQRKPGQPPKKLIYWASTRESGVPDRFTGSGSGTDFTLTISGVQAEDLAVYYCQNNYFYPLTFGAGTKLELK
实施例2 抗人CLDN18.2嵌合抗体的制备
将鼠源抗人CLDN18.2单克隆抗体的重链可变区序列和公开发表的人单克隆抗体IgG1亚类的重链恒定区序列(参见SEQ ID NO:48)拼接在一起,构建到哺乳动物细胞表达载体中;将鼠源抗人CLDN18.2单克隆抗体的轻链可变区序列和公开发表的人单克隆抗体κ亚类的轻链恒定区序列(参见SEQ ID NO:49)拼接在一起,构建到哺乳动物细胞表达载体中。构建好的抗人CLDN18.2嵌合抗体的重链载体和轻链载体配对混合,使用聚乙烯亚胺(PEI)转染HEK293细胞,约7天后收集细胞上清,使用ProteinA纯化得到抗人CLDN18.2嵌合抗体蛋白。
嵌合抗体命名为“鼠抗简称-xiIgG”。
实施例3 抗人CLDN18.2嵌合抗体的体外细胞结合实验
将抗人CLDN18.2嵌合抗体从100nM的起始浓度开始做2倍的梯度倍比稀释,共16个浓度点,各个浓度点的抗体取10ul加入384孔板。100g室温离心5分钟收集细胞表面表达CLDN18.2的CHOK1细胞,用含0.5%BSA的PBS洗涤细胞一次,100g室温离心5分钟,重悬细胞为密度约2x106个细胞/ml,取10ul加入到已加抗体的384孔板的孔中。4℃孵育1小时后,加入荧光标记的羊抗人IgG二抗。继续于4℃孵育1小时后,用流式细胞仪分析细胞群的平均荧光读值。
结果显示嵌合抗体与表达人CLDN18.2细胞有nM级别的特异结合。
实施例4 抗人CLDN18.2鼠抗的人源化
综合Kabat、Chothia的抗体编码方案,确定鼠抗的重链和轻链中6个抗原互补决定簇(CDR)的氨基酸序列区域及支撑抗体保守三维构象的框架区域(framework region)。随后通过分析搜索已知人源抗体序列,选择与鼠抗最为相似接近的人源抗体重链可变区序列,选择其抗体框架区序列作为模板,将鼠抗重链CDR与人源抗体框架区结合,最终生成人源化抗体重链可变区序列。同样过程,生成人源化抗体轻链可变区序列。
鼠抗CDR直接移植至人框架区的抗体常出现结合活性急剧下降,因此需要将框架区个别氨基酸从人源的改回鼠源的。确定回复突变位点,一是对照设计好的人源化抗体序列和原始的鼠抗序列,检查有哪些氨基酸不同;二是检查这些氨基酸是否对支持抗体结构起重要作用或者对与抗原的结合起重要作用。同时需要检查人源化设计后的序列是否有一些潜在的翻译后修饰位点(PTM),如N(天冬酰胺)糖基化位点、N脱酰胺化位点、D(天冬氨酸)异构化位点等。
基于上述8K13_vh、8K13_vl、16K15_vh和16K15_vl的人源化版本示例如下:
鼠抗8k13的人源化
重链可变区
>8K13_vh_hz0(SEQ ID NO:5;CDR按顺序为:SEQ ID NO:23、SEQ ID NO:24、SEQ IDNO:25)
QVQLVQSGAEVKKPGSSVKVSCKASGYAFSNYWMNWVRQAPGQGLEWMGQIYPGNGDTKYSGKFNSRVTITADESTSTAYMELSSLRSEDTAVYYCARFYYGNVMDYWGQGTLVTVSS
>8K13_vh_hz1(SEQ ID NO:6;CDR按顺序为:SEQ ID NO:23、SEQ ID NO:24、SEQ IDNO:25)
QVQLVQSGAEVKKPGSSVKVSCKASGYAFSNYWMNWVRQAPGQGLEWMGQIYPGNGDTKYSGKFNSRVTITADKSTNTAYMELSSLRSEDTAVYYCARFYYGNVMDYWGQGTLVTVSS
>8K13_vh_hz2(SEQ ID NO:7;CDR按顺序为:SEQ ID NO:23、SEQ ID NO:26、SEQ IDNO:25)
QVQLVQSGAEVKKPGSSVKVSCKASGYAFSNYWMNWVRQAPGQGLEWMGQIYPGSGDTKYSGKFQSRVTITADKSTNTAYMELSSLRSEDTAVYYCARFYYGNVMDYWGQGTLVTVSS
>8K13_vh_hz3(SEQ ID NO:8;CDR按顺序为:SEQ ID NO:23、SEQ ID NO:26、SEQ IDNO:25)
EVQLVQSGAEVKKPGSSVKVSCKASGYAFSNYWMNWVRQAPGQGLEWMGQIYPGSGDTKYSGKFQSRVTITADKSTSTAYMELSSLRSEDTAVYYCARFYYGNVMDYWGQGTLVTVSS
轻链可变区
>8K13_vl_hz0(SEQ ID NO:9;CDR按顺序为:SEQ ID NO:29、SEQ ID NO:30、SEQ IDNO:31)
DIVMTQSPDSLAVSLGERATINCKSSQTLLNGGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNGYSYPLTFGQGTKLEIK
>8K13_vl_hz1(SEQ ID NO:10;CDR按顺序为:SEQ ID NO:29、SEQ ID NO:30、SEQID NO:31)
DIVLTQSPDSLAVSLGERATINCKSSQTLLNGGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDLAVYYCQNGYSYPLTFGQGTKLEIK
>8K13_vl_hz4(SEQ ID NO:11;CDR按顺序为:SEQ ID NO:32、SEQ ID NO:30、SEQID NO:33)
DIVLTQSPDSLAVSLGERATINCKSSQTLLSGGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDLAVYYCQQGYSYPLTFGQGTKLEIK
>8K13_vl_hz5(SEQ ID NO:12;CDR按顺序为:SEQ ID NO:32、SEQ ID NO:30、SEQID NO:33)
DIVLTQSPDSLAVSLGERATINCKSSQTLLSGGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQGYSYPLTFGQGTKLEIK
鼠抗16k15的人源化
重链可变区
>16k15_vh_hz0(SEQ ID NO:13;CDR按顺序为:SEQ ID NO:34、SEQ ID NO:35、SEQID NO:36)
EVQLVESGGGLVQPGGSLRLSCAASGITLSTYAMSWVRQAPGKGLEWVSSIISGGITYYLDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARKYHGNALDYWGQGTLVTVSS
>16k15_vh_hz1(SEQ ID NO:14;CDR按顺序为:SEQ ID NO:34、SEQ ID NO:35、SEQID NO:36)
EVQLVESGGGLVQPGGSLRLSCAGSGITLSTYAMSWVRQAPGKGLEWVSSIISGGITYYLDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARKYHGNALDYWGQGTLVTVSS
>16k15_vh_hz2(SEQ ID NO:15;CDR按顺序为:SEQ ID NO:34、SEQ ID NO:35、SEQID NO:36)
EVQLVESGGGLVQPGGSLRLSCAGSGITLSTYAMSWVRQAPGKGLEWVSSIISGGITYYLDSVKGRFTISRDNAKNTLYLQMSSLRAEDTAVYYCARKYHGNALDYWGQGTLVTVSS
>16k15_vh_hz3(SEQ ID NO:16;CDR按顺序为:SEQ ID NO:34、SEQ ID NO:35、SEQID NO:36)
EVQLVESGGGLVQPGGSLRLSCAGSGITLSTYAMSWVRQAPGKGLEWVSSIISGGITYYLDSVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARKYHGNALDYWGQGTLVTVSS
轻链可变区
>16K15_vl_hz0(SEQ ID NO:17;CDR按顺序为:SEQ ID NO:39、SEQ ID NO:30、SEQID NO:40)
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQRNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNNYFYPLTFGQGTKLEIK
>16K15_vl_hz1(SEQ ID NO:18;CDR按顺序为:SEQ ID NO:39、SEQ ID NO:30、SEQID NO:40)
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQRNYLTWYQQKPGQPPKKLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDLAVYYCQNNYFYPLTFGQGTKLEIK
>16K15_vl_hz2(SEQ ID NO:19;CDR按顺序为:SEQ ID NO:41、SEQ ID NO:30、SEQID NO:40)
DIVMTQSPDSLAVSLGERATINCKSSQSLLSSGNQRNYLTWYQQKPGQPPKKLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDLAVYYCQNNYFYPLTFGQGTKLEIK
>16K15_vl_hz3(SEQ ID NO:20;CDR按顺序为:SEQ ID NO:41、SEQ ID NO:30、SEQID NO:42)
DIVMTQSPDSLAVSLGERATINCKSSQSLLSSGNQRNYLTWYQQKPGQPPKKLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDLAVYYCQSNYFYPLTFGQGTKLEIK
>16K15_vl_hz4(SEQ ID NO:21;CDR按顺序为:SEQ ID NO:41、SEQ ID NO:30、SEQID NO:43)
DIVMTQSPDSLAVSLGERATINCKSSQSLLSSGNQRNYLTWYQQKPGQPPKKLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDLAVYYCQQNYFYPLTFGQGTKLEIK
>16k15_vl_hz5(SEQ ID NO:22;CDR按顺序为:SEQ ID NO:41、SEQ ID NO:30、SEQID NO:43)
DIVMTQSPDSLAVSLGERATINCKSSQSLLSSGNQRNYLTWYQQKPGQPPKKLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQNYFYPLTFGQGTKLEIK
将人源化抗体重链可变区基因构建到含人单克隆抗体IgG1亚类的重链恒定区序列(参见SEQ ID NO:48)的哺乳动物细胞表达载体中;轻链可变区基因构建到含人单克隆抗体κ亚类的轻链恒定区序列(参见SEQ ID NO:49)的哺乳动物细胞表达载体中。构建好的抗人CLDN18.2人源化抗体的重链载体和轻链载体配对混合,使用聚乙烯亚胺(PEI)转染HEK293细胞,约7天后收集细胞上清,使用ProteinA纯化得到抗人CLDN18.2人源化抗体蛋白。
本发明的人源化抗体命名为“鼠抗简称-hz”。鼠抗CDR直接移植至人框架区的抗体命名为“鼠抗简称-hz00”。进一步改造得到的抗体根据序列不同进行编号,例如,抗体8k13-hz11的编号来自其重链和轻链可变区的编号8K13_vh_hz1和8K13_vl_hz1。
利用Fortebio(BLITZ pro1.1.0.28)仪器分析嵌合抗体及其人源化抗体与抗原人CLDN18.2的结合动力学参数。测定前先将NTA生物探针浸泡于PBS中10分钟;然后将该探针置于含100nM的抗原的PBS中300秒,捕获带His标签的抗原;进一步将探针与100nM抗体进行结合反应,结合时间400秒;之后将探针转移至PBS中,进行解离反应,时间为600秒。实验完毕,扣除空白对照响应值,用软件进行1:1Langmuir结合模式拟合,计算抗原抗体结合的动力学常数。结果见表2。
表2.鼠抗人源化后的结合动力学参数比较
实施例5 抗人CLDN18.2人源化抗体的体外结合亲和力的动力学实验
采用GE公司BIAcore仪器S200测定抗体抗原相互作用力。
参考GE公司Human antibody capture kit(货号BR-1008-39,Lot10261753)操作说明,首先在传感芯片CM5分析通道和对照样品通道都饱和偶联最大量抗人Fc抗体,然后在分析通道流过含有7.5μg/ml抗人CLDN18.2嵌合抗体、人源化抗体或IMAB362的缓冲液使其均匀分布,最后在分析通道和对照样品通道一起流过梯度稀释的抗原样品(起始浓度20nM,1:3稀释8个浓度点,并且设定0.741nm浓度点重复),测定抗体抗原结合后发生的光反应值。经仪器软件拟合分析,最终得到抗体的结合常数Kon和解离常数Koff,以及亲和力常数KD。
结果表明,抗人CLDN18.2人源化抗体的体外结合亲和力常数与原始鼠抗比没有显著的改变,见表3。
表3.抗体的结合动力学
抗体 | ka(M-1s-1) | kd(s-1) | KD(M) |
8k13-xiIgG | 8.19E+04 | 5.73E-04 | 6.99E-09 |
8k13-hz24 | 7.09E+04 | 2.02E-03 | 2.85E-08 |
16k15-xiIgG | 1.07E+05 | 1.44E-03 | 1.35E-08 |
16k15-hz22 | 5.71E+04 | 5.63E-04 | 9.85E-09 |
IMAB362 | 1.48E+06 | 0.201 | 1.36E-07 |
实施例6 抗人CLDN18.2人源化抗体的体外细胞学试验
6.1补体依赖性细胞毒性(CDC)
使用Quidel公司的商品化人血清全补体对本发明人源化抗体进行分析,分析其对稳定表达人CLDN18.2的CHOK1、BxPC3和NCI-N87细胞诱导补体依赖性细胞毒性(CDC)的能力。
将细胞与终浓度为250μg/ml至3.8ng/ml的待测抗体混匀;加入溶于细胞培养基RPMI-1640中的6.25%人血清全补体,在37℃下孵育3小时;然后通过CCK-8试剂盒进行细胞毒性检测;最终通过MD酶标仪检测450nm吸光度。通过吸光值采用softmax pro7软件进行4参数拟合曲线,计算样品的EC50值。
结果表明,抗人CLDN18.2人源化抗体针对靶点表达细胞有特异的补体依赖性细胞毒(CDC)活性,并且细胞杀伤活性显著优于IMAB362,见表4。
表4.抗体的CDC
靶点表达细胞 | a.CHOK1 | b.BxPC3 | c.NCI-N87 |
抗体 | EC50(nM) | EC50(nM) | EC50(nM) |
8k13-hz24 | 21.27 | 28.53 | 57.7 |
16k15-hz22 | 5.2 | 12.22 | 17.36 |
IMAB362 | 30.45 | 52.23 | 474.2 |
6.2抗体依赖性细胞毒性(ADCC)
使用工程改造的Jurkat细胞作为效应细胞,该细胞稳定表达FcγRIIIa-FcεRIaγ杂合受体,由NFAT应答元件驱动表达萤火虫萤光素酶。抗体在ADCC作用机制中的生物活性通过NFAT通路活化产生的萤光素酶定量。将1.5E5个效应细胞与终浓度33μg/ml至85pg/ml的待测抗体混匀,然后将2.5E4个靶细胞加入其中(效应细胞与靶细胞E:T比例6:1),在37℃下孵育16小时;然后通过Promega公司试剂盒Bio-GloTMLuciferase Assay System进行检测;最终通过MD酶标仪检测LUM值。
数据处理方式如下,诱导倍数=(受检孔读值–背景值)/(阴性对照孔读值–背景值),用prism软件进行4参数拟合曲线,计算出样品的EC50值。
结果表明,抗人CLDN18.2人源化抗体针对靶点表达细胞有特异的抗体依赖性细胞毒(ADCC)活性,细胞杀伤活性与IMAB362相当,见表5。
表5.抗体的ADCC
靶点表达细胞 | a.CHOK1 | b.BxPC3 | c.NCI-N87 |
抗体 | EC50(nM) | EC50(nM) | EC50(nM) |
8k13-hz24 | 0.5894 | 0.3045 | 0.1499 |
16k15-hz22 | 0.7122 | 0.1759 | 0.0677 |
IMAB362 | 0.5929 | 0.1056 | 0.0909 |
实施例7 抗人CLDN18.2人源化抗体的表位研究
使用表位竞争法对本发明的抗体及对照抗体的表位进行研究。
制备Rituximab(无关抗体,即对照抗体)、8k13-hz35(SEQ ID NO:8+SEQ ID NO:12)和16k15-hz35(SEQ ID NO:16+SEQ ID NO:22)的Fab形式抗体Isotype-Fab、8k13-hz35-Fab和16k15-hz35-Fab。然后,首先将竞争抗体IMAB362稀释为0.05ug/ml。用含竞争抗体的溶液将待分析的8k13-hz35-Fab、16k15-hz35-Fab和Isotype-Fab稀释到200nM的浓度,做2倍的梯度倍比稀释,共8个浓度点。各个浓度点的抗体取10ul加入384孔板。100g室温离心5分钟收集细胞表面表达CLDN18.2 CHOK1细胞,用含0.5%BSA的PBS洗涤细胞一次,100g室温离心5分钟。重悬细胞为密度约2x106个细胞/毫升,取10ul加入到已加抗体的384孔板的孔中,4℃孵育过夜。第二天加入1:500稀释的荧光标记羊抗人IgG二抗10ul。于4℃孵育1小时后,用流式细胞仪分析细胞群的平均荧光读值。
结果见图1。可知,本发明的抗体8k13-hz35与IMAB362结合不同的抗原表位,而16k15-hz35与IMAB362存在抗原识别表位的竞争关系。
实施例8 FACS检测抗人CLDN18.2人源化抗体与CHO细胞表面人CLDN18.2重组蛋白的结合活性
将重组表达人CLDN18.2的CHO细胞(CHO/CLDN18.2)悬液分别与人源化抗体(8k13-hz35、16k15-hz35、8k13-hz24、16k15-hz22)(浓度为5μg/mL起始3倍连续稀释5个梯度)在4℃孵育60min,并设阴性对照(NC)IgG1同型对照抗体NC-huIgG1。以PBS洗涤细胞3次后,加入1:200稀释的羊抗人IgG-FITC(Cat:F9512,Sigma)并4℃孵育45min。PBS洗涤细胞3次后通过流式细胞仪(型号B49007AD,SNAW31211,BECKMAN COULTER)检测细胞的平均荧光强度(MFI),以检测人源化抗体与CHO细胞表面的人CLDN18.2结合能力。
结果如图2所示,抗体8k13-hz35、16k15-hz35可以有效结合CHO细胞表面的人CLDN18.2重组蛋白,且结合能力与抗体8k13-hz24、16k15-hz22结合能力相当。
实施例9 抗人CLDN18.2人源化抗体结合细胞表面CLDN18.2的内化活性
将重组表达人CLDN18.2的NCI-N87 CLDN18.2细胞(Cat.:KC-1222,康源博创),以2×103个细胞/孔的密度接种于96孔细胞培养板,培养24小时。弃上清,用RPMI 1640(含10%FBS),37℃封闭60min,离心,RPMI1640(含10%FBS)洗涤2遍;将使用Mix-n-StainTMCFTM488A(Cat:MX488AS100,Sigma)标记的人源化抗体8k13-hz35、16k15-hz35、对照抗体IMAB362、内化阳性抗体Sacituzumab以及IgG同型对照抗体NC-huIgG1,用RPMI 1640(含10%FBS)稀释至1μg/mL,加入NCI-N87 CLDN18.2细胞中,100ul/孔。每个样品分为2组,一组在37℃下孵育作为内吞组,一组在4℃下孵育做为阴性对照;阴性对照孵育1小时后,4℃离心并PBS洗涤2次,用荧光显微镜观察并拍照,37℃内吞组孵育4小时后,室温离心并PBS洗涤2次,用荧光显微镜观察并拍照。
结果见图3,表明人源化抗体8k13-hz35和16k15-hz35在37℃条件下都能够被CLDN18.2介导内吞,在细胞质内呈点状分布。提示抗体人源化后仍然能够保持内化活性。
(二)抗体药物偶联物的制备与表征
以下实施例中采用的药物接头包括:
MC-GGFG-Dxd,喜树碱类拓扑异构酶I抑制剂-ADC接头,本发明上下文中缩写为“EX”。
CL2A-SN38,喜树碱类拓扑异构酶I抑制剂-ADC接头,本发明上下文中缩写为“EX2”。
MC-VC-MMAE,MMAE微管抑制剂-ADC接头,本发明上下文中缩写为“M”。
L-A,苯并氮卓类TLR8激动剂-ADC接头,本发明上下文中缩写为“TLR8”。
L-B,环二核苷酸类STING激动剂-ADC接头,本发明上下文中缩写为“STING”。
示例性地,采用了以下实验方法:
1.抗体的还原
用含有50mM氯化钠和2mM EDTA的磷酸盐缓冲液(50mM,pH6.5;简称为PBS6.5/EDTA)制备抗体溶液(抗体浓度为10mg/ml),向其中加入TCEP(Sigma-Aldrich)水溶液,使TCEP为抗体的2-10倍摩尔当量,然后在25℃孵育2h。
2.抗体与药物接头的偶联
使用PBS6.5/EDTA,将还原后抗体浓度调整为5mg/ml。加入N,N-二甲基乙酰胺(DMA),然后加入药物接头的二甲基亚砜溶液(药物接头浓度为10mg/ml),使药物接头为抗体的6-12摩尔当量,然后加入药物接头的二甲基亚砜溶液(药物接头浓度为10mg/ml),使药物接头为抗体的5-12倍摩尔当量,且将DMA在整个反应体系中的体积比控制为10-20%。将该溶液在室温条件下继续搅拌30-60分钟,使药物接头与抗体偶联。
3.抗体药物偶联物的纯化
用磷酸缓冲液(pH7.4,浓度为50mM;简称为PB7.4)平衡NAP-25色谱柱,针对一根该NAP-25柱,装填一定量的抗体药物偶联反应液,然后分离获得用PB7.4洗脱的组分。使用Amicon Ultra-15(30000MWCO,Millipore Corporation)浓缩,除菌过滤。
使用如下抗体药物偶联物浓度测定方法测定偶联样品中抗体药物偶联物的浓度。
4.抗体药物偶联物浓度测定方法
4.1抗体药物偶联物浓度测定方法-EX
样品中的连接药物浓度可通过测定在280nm波长下的UV吸光度,然后进行下述计算而得到。
由于某一波长下的总吸光度等于存在于体系内的所有吸收化学物质种类的吸光度的和(吸光度的加成性),所以,假设在抗体与药物接头偶联前后,抗体及药物接头的摩尔吸光系数不发生变化时,抗体药物偶联物中的抗体浓度及药物浓度如下述的关系式所示。
4.2抗体-药物偶联物浓度测定方法-TLR8
如上,抗体药物偶联物中的抗体浓度及药物浓度如下述的关系式所示。
4.3抗体-药物偶联物浓度测定方法-STING
如上抗体药物偶联物中的抗体浓度及药物浓度如下述的关系式所示。
4.4抗体-药物偶联物浓度测定方法-M
如上,抗体药物偶联物中的抗体浓度及药物浓度如下述的关系式所示。
4.5抗体-药物偶联物浓度测定方法-EX2
如上,抗体药物偶联物中的抗体浓度及药物浓度如下述的关系式所示。
5.抗体药物偶联物中每一分子抗体的药物平均连接数(DAR)的测定方法
5.1疏水作用色谱法
HPLC分析用样品制备:以流动相B为稀释液,将抗体药物偶联物稀释至2mg/ml,0.22μM滤头过滤,用于HPLC分析。
DAR计算公式:
DAR=∑(加权峰面积)/100,即DAR=(D0峰面积比*0+D1峰面积比*1+D2峰面积比*2+D3峰面积比*3+D4峰面积比*4+D5峰面积比*5+D6峰面积比*6+D7峰面积比*7+D8峰面积比*8)/100。
5.1.1在下述的测定条件下进行HIC分析
HPLC仪器:Waters Acquity Arc
流动相A:2.5M硫酸铵+125mM PB,pH 6.8
流动相B:125mM PB,pH 6.8
流动相C:IPA
流动相D:水
分析柱:Tosoh,TSKgel Butyl-NPR;4.6*100mm;2.5μm,PN 0042168
进样体积:20μL
流速:0.5mL/min
柱温:25℃
检测器:PDA检测器
检测波长:280nm
梯度:
时间(min) | 流动相A | 流动相B | 流动相C | 流动相D |
0 | 60 | 0 | 5 | 35 |
15 | 0 | 50 | 5 | 45 |
20 | 0 | 50 | 5 | 45 |
20.1 | 60 | 0 | 5 | 35 |
35 | 60 | 0 | 5 | 35 |
5.1.2在下述的测定条件下进行HIC分析
HPLC仪器:Waters Acquity Arc
流动相A:2.5M硫酸铵+125mM PB,pH 6.8
流动相B:125mM PB,pH 6.8
流动相C:IPA
流动相D:水
分析柱:Tosoh,TSKgel Ether-5PW;7.5*75mm;2.5μm,PN 0008641
进样体积:50μL
流速:0.6mL/min
柱温:25℃
检测器:PDA检测器
检测波长:280nm
梯度:
时间(min) | 流动相A | 流动相B | 流动相C | 流动相D |
0 | 60 | 0 | 5 | 35 |
30 | 0 | 50 | 5 | 45 |
33 | 0 | 50 | 5 | 45 |
33.1 | 60 | 0 | 5 | 35 |
45 | 60 | 0 | 5 | 35 |
5.2反相高效液相色谱法(RPLC)
HPLC分析用样品制备:将抗体药物偶联物(约1mg/mL,60μL)加入至二硫苏糖醇水溶液(100mM,15μL)中,混匀,37℃下孵育30分钟,用于HPLC分析。
DAR计算公式:
DAR=2*(∑轻链加权峰面积+∑重链加权峰面积)/100.即:DAR=2*(L0峰面积比*0+L1峰面积比*1+H0峰面积比*0+H1峰面积比*1+H2峰面积比*2+H3峰面积比*3)/100
5.2.1在下述的测定条件下进行RPLC分析
HPLC仪器:Waters Acquity Arc
流动相A:ACN+0.1%TFA
流动相B:H2O+0.1%TFA
进样体积:15μL
流速:0.25mL/min
柱温:80℃
检测器:PDA检测器
检测波长:280nm
梯度:
5.2.2在下述的测定条件下进行RPLC分析
HPLC仪器:Waters Acquity Arc
流动相A:ACN+0.1%TFA
流动相B:H2O+0.1%TFA
进样体积:5μL
流速:0.25mL/min
柱温:80℃
检测器:PDA检测器
检测波长:280nm
梯度:
时间(min) | ACN | H2O |
0 | 29 | 71 |
3 | 33 | 64 |
12.5 | 42 | 58 |
20 | 55 | 45 |
20.1 | 29 | 71 |
30 | 29 | 71 |
6.抗体药物偶联物分子大小异质性测定方法
在下述的测定条件下进行SEC分析:
HPLC分析用样品制备:以流动相为稀释液,将抗体药物偶联物稀释至2mg/ml,0.22μM滤头过滤,用于HPLC分析。
HPLC仪器:Waters Acquity Arc
流动相:100mM PB+200mM Arg·HCl+5%IPA,pH 6.8
进样体积:10μL
流速:0.6mL/min
柱温:30℃
检测器:PDA检测器
检测波长:280nm
梯度:等度洗脱
实施例组1 药物接头的合成
实施例1 苯并氮卓类TLR8激动剂-ADC接头的合成
按照专利公开文件CN110612104A所述方法合成TLR8,其为白色固体,LC-MS(ESI):Pos(M+1)=767。
实施例2 环二核苷酸类STING激动剂-ADC接头的合成
按照专利公开文件WO2018/100558A2所述方法合成STING,其为淡黄色固体,LC-MS(ESI):Pos(M+1)=1164。
实施例组2 抗体药物偶联物的制备
实施例1 抗体药物偶联物1(8k13-hz35-TLR8-1103)
(1)抗体的还原
采用上文实验方法部分的“1.抗体的还原”中所述操作,将抗体8k13-hz35还原(TCEP用量为抗体的2.0eq),得到还原后的抗体溶液。
(2)抗体与药物接头的偶联
采用上文实验方法部分的“2.抗体与药物接头的偶联”中所述操作,向所得还原液中加入TLR8的二甲基亚砜溶液(TLR8的用量为抗体的5.0eq),反应得到含有标题抗体药物偶联物的溶液。
(3)抗体药物偶联物的纯化
采用上文实验方法部分的“3.抗体药物偶联物的纯化”中所述操作,将上述溶液纯化,得到含有标题抗体药物偶联物的溶液。
(4)抗体药物偶联物分子特性评价
采用上文实验方法部分的“4.2抗体-药物偶联物浓度测定方法-TLR8”中所述操作,测得溶液中抗体药物偶联物的浓度为6.3mg/mL。
采用上文实验方法部分的“5.抗体药物偶联物中每一分子抗体的药物平均连接数(DAR)的测定方法”第5.1.2节中所述操作,测得药物平均连接数(DAR)为4.63(图4)。
采用上文实验方法部分的“6.抗体药物偶联物分子大小异质性测定方法”中所述操作,测得主峰含量为98.72%(图5)。如图5所示,采用分子排阻色谱法分析分子大小异质性,除主峰外,高分子质量物质和抗体偶联药物中的碎片含量也能检测。
实施例2 抗体药物偶联物2(8k13-hz35-STING-D4-1203)
(1)抗体的还原
采用上文实验方法部分的“1.抗体的还原”中所述操作,将抗体8k13-hz35还原(TCEP用量为抗体的2.1eq),得到还原后的抗体溶液。
(2)抗体与药物接头的偶联
采用上文实验方法部分的“2.抗体与药物接头的偶联”中所述操作,向所得还原液中加入STING的二甲基亚砜溶液(STING用量为抗体的5.8eq),反应得到含有标题抗体药物偶联物的溶液。
(3)抗体药物偶联物的纯化
同实施例1。
(4)抗体药物偶联物分子特性评价
采用上文实验方法部分的“4.3抗体-药物偶联物浓度测定方法-STING”中所述操作,测得溶液中抗体药物偶联物的浓度为7.7mg/mL。
采用上文实验方法部分的“5.抗体药物偶联物中每一分子抗体的药物平均连接数(DAR)的测定方法”第5.2.2节中所述操作,测得药物平均连接数(DAR)为4.02(图6)。
采用上文实验方法部分的“6.抗体药物偶联物分子大小异质性测定方法”中所述操作,测得主峰含量为85.15%(图7)。
实施例3 抗体药物偶联物3(8k13-hz35-M-20200318-1)
(1)抗体的还原
采用上文实验方法部分的“1.抗体的还原”中所述操作,将抗体8k13-hz35还原(TCEP用量为抗体的2.2eq),得到还原后的抗体溶液。
(2)抗体与药物接头的偶联
采用上文实验方法部分的“2.抗体与药物接头的偶联”中所述操作,向所得还原液中加入M的二甲基亚砜溶液(M用量为抗体的5.5eq),反应得到含有标题抗体药物偶联物的溶液。
(3)抗体药物偶联物的纯化
同实施例1。
(4)抗体药物偶联物分子特性评价
采用上文实验方法部分的“4.4抗体-药物偶联物浓度测定方法-M”中所述操作,测得溶液中抗体药物偶联物的浓度为7.7mg/mL。
采用上文实验方法部分的“5.抗体药物偶联物中每一分子抗体的药物平均连接数(DAR)的测定方法”第5.1.1节中所述操作,测得药物平均连接数(DAR)为4.04(图8)。
采用上文实验方法部分的“6.抗体药物偶联物分子大小异质性测定方法”中所述操作,测得主峰含量为99.64%(图9)。
实施例4 抗体药物偶联物4(8k13-hz35-EX-DAR4-20200922)
(1)抗体的还原
采用上文实验方法部分的“1.抗体的还原”中所述操作,将抗体8k13-hz35还原(TCEP用量为抗体的2.1eq),得到还原后的抗体溶液。
(2)抗体与药物接头的偶联
采用上文实验方法部分的“2.抗体与药物接头的偶联”中所述操作,向所得还原液中加入EX的二甲基亚砜溶液(EX用量为抗体的5.2eq),反应得到含有标题抗体药物偶联物的溶液。
(3)抗体药物偶联物的纯化
同实施例1。
(4)抗体药物偶联物分子特性评价
采用上文实验方法部分的“4.1抗体-药物偶联物浓度测定方法-EX”中所述操作,测得溶液中抗体药物偶联物的浓度为7.7mg/mL。
采用上文实验方法部分的“5.抗体药物偶联物中每一分子抗体的药物平均连接数(DAR)的测定方法”第5.2.1节中所述操作,测得药物平均连接数(DAR)为3.53(图10)。
采用上文实验方法部分的“6.抗体药物偶联物分子大小异质性测定方法”中所述操作,测得主峰含量为99.80%(图11)。
实施例5 抗体药物偶联物5(8k13-hz35-EX-DAR8-20200421)
(1)抗体的还原
采用上文实验方法部分的“1.抗体的还原”中所述操作,将抗体8k13-hz35还原(TCEP用量为抗体的10eq),得到还原后的抗体溶液。
(2)抗体与药物接头的偶联
采用上文实验方法部分的“2.抗体与药物接头的偶联”中所述操作,向所得还原液中加入EX的二甲基亚砜溶液(EX用量为抗体的11eq),反应得到含有标题抗体药物偶联物的溶液。
(3)抗体药物偶联物的纯化
同实施例1。
(4)抗体药物偶联物分子特性评价
采用上文实验方法部分的“4.1抗体-药物偶联物浓度测定方法-EX”中所述操作,测得溶液中抗体药物偶联物的浓度为7.7mg/mL。
采用上文实验方法部分的“5.抗体药物偶联物中每一分子抗体的药物平均连接数(DAR)的测定方法”第5.1.1节中所述操作,测得药物平均连接数(DAR)为7.83(图12)。
采用上文实验方法部分的“6.抗体药物偶联物分子大小异质性测定方法”中所述操作,测得主峰含量为98.81%(图13)。
实施例6 抗体药物偶联物6(8k13-hz35-EX2-DAR4-20200922)
(1)抗体的还原
采用上文实验方法部分的“1.抗体的还原”中所述操作,将抗体8k13-hz35还原(TCEP用量为抗体的2.2eq),得到还原后的抗体溶液。
(2)抗体与药物接头的偶联
采用上文实验方法部分的“2.抗体与药物接头的偶联”中所述操作,向所得还原液中加入EX2的二甲基亚砜溶液(EX用量为抗体的5.2eq),反应得到含有标题抗体药物偶联物的溶液。
(3)抗体药物偶联物的纯化
同实施例1。
(4)抗体药物偶联物分子特性评价
采用上文实验方法部分的“4.5抗体-药物偶联物浓度测定方法-EX2”中所述操作,测得溶液中抗体药物偶联物的浓度为4.5mg/mL。
采用上文实验方法部分的“5.抗体药物偶联物中每一分子抗体的药物平均连接数(DAR)的测定方法”第5.2.2节中所述操作,测得药物平均连接数(DAR)为3.41(图14)。
采用上文实验方法部分的“6.抗体药物偶联物分子大小异质性测定方法”中所述操作,测得主峰含量为99.85%(图15)。
实施例7 抗体药物偶联物7(8k13-hz35-EX2-DAR8-20200922)
(1)抗体的还原
采用上文实验方法部分的“1.抗体的还原”中所述操作,将抗体8k13-hz35还原(TCEP用量为抗体的10.1eq),得到还原后的抗体溶液。
(2)抗体与药物接头的偶联
采用上文实验方法部分的“2.抗体与药物接头的偶联”中所述操作,向所得还原液中加入EX2的二甲基亚砜溶液(EX用量为抗体的12eq),反应得到含有标题抗体药物偶联物的溶液。
(3)抗体药物偶联物的纯化
同实施例1。
(4)抗体药物偶联物分子特性评价
采用上文实验方法部分的“4.5抗体-药物偶联物浓度测定方法-EX2”中所述操作,测得溶液中抗体药物偶联物的浓度为4.4mg/mL。
采用上文实验方法部分的“5.抗体药物偶联物中每一分子抗体的药物平均连接数(DAR)的测定方法”第5.2.1节中所述操作,测得药物平均连接数(DAR)为7.12(图16)。
采用上文实验方法部分的“6.抗体药物偶联物分子大小异质性测定方法”中所述操作,测得主峰含量为99.71%(图17)。
实施例8 抗体药物偶联物8(16k15-hz35-M-20200318-1)
(1)抗体的还原
采用上文实验方法部分的“1.抗体的还原”中所述操作,将抗体16k15-hz35还原(TCEP用量为抗体的2.3eq),得到还原后的抗体溶液。
(2)抗体与药物接头的偶联
采用上文实验方法部分的“2.抗体与药物接头的偶联”中所述操作,向所得还原液中加入M的二甲基亚砜溶液(M用量为抗体的5.3eq),反应得到含有标题抗体药物偶联物的溶液。
(3)抗体药物偶联物的纯化
同实施例1。
(4)抗体药物偶联物分子特性评价
采用上文实验方法部分的“4.4抗体-药物偶联物浓度测定方法-M”中所述操作,测得溶液中抗体药物偶联物的浓度为7.7mg/mL。
采用上文实验方法部分的“5.抗体药物偶联物中每一分子抗体的药物平均连接数(DAR)的测定方法”第5.1.1节中所述操作,测得药物平均连接数(DAR)为4.03(图18)。
采用上文实验方法部分的“6.抗体药物偶联物分子大小异质性测定方法”中所述操作,测得主峰含量为99.18%(图19)。
实施例9 抗体药物偶联物9(16k15-hz35-EX-20200421)
(1)抗体的还原
采用上文实验方法部分的“1.抗体的还原”中所述操作,将抗体16k15-hz35还原(TCEP用量为抗体的9.9eq),得到还原后的抗体溶液。
(2)抗体与药物接头的偶联
采用上文实验方法部分的“2.抗体与药物接头的偶联”中所述操作,向所得还原液中加入EX的二甲基亚砜溶液(EX用量为抗体的11.5eq),反应得到含有标题抗体药物偶联物的溶液。
(3)抗体药物偶联物的纯化
同实施例1。
(4)抗体药物偶联物分子特性评价
采用上文实验方法部分的“4.1抗体-药物偶联物浓度测定方法-EX”中所述操作,测得溶液中抗体药物偶联物的浓度为7.7mg/mL。
采用上文实验方法部分的“5.抗体药物偶联物中每一分子抗体的药物平均连接数(DAR)的测定方法”第5.1.1节中所述操作,测得药物平均连接数(DAR)为7.92(图20)。
采用上文实验方法部分的“6.抗体药物偶联物分子大小异质性测定方法”中所述操作,测得主峰含量为98.39%(图21)。
实施例10 抗体药物偶联物10(IMAB362-M-1)
(1)抗体的还原
采用上文实验方法部分的“1.抗体的还原”中所述操作,将抗体IMAB362还原(TCEP用量为抗体的2.2eq),得到还原后的抗体溶液。
(2)抗体与药物接头的偶联
采用上文实验方法部分的“2.抗体与药物接头的偶联”中所述操作,向所得还原液中加入M的二甲基亚砜溶液(M用量为抗体的5.2eq),反应得到含有标题抗体药物偶联物的溶液。
(3)抗体药物偶联物的纯化
同实施例1。
(4)抗体药物偶联物分子特性评价
采用上文实验方法部分的“4.4抗体-药物偶联物浓度测定方法-M”中所述操作,测得溶液中抗体药物偶联物的浓度为6.9mg/mL。
采用上文实验方法部分的“5.抗体药物偶联物中每一分子抗体的药物平均连接数(DAR)的测定方法”第5.1.1节中所述操作,测得药物平均连接数(DAR)为3.28(图22)。
采用上文实验方法部分的“6.抗体药物偶联物分子大小异质性测定方法”中所述操作,测得主峰含量为90.13%(图23)。
实施例11 抗体药物偶联物11(IMAB362-M-2)
(1)抗体的还原
采用上文实验方法部分的“1.抗体的还原”中所述操作,将抗体IMAB362还原(TCEP用量为抗体的8eq),得到还原后的抗体溶液。
(2)抗体与药物接头的偶联
采用上文实验方法部分的“2.抗体与药物接头的偶联”中所述操作,向所得还原液中加入M的二甲基亚砜溶液(M用量为抗体的10eq),反应得到含有标题抗体药物偶联物的溶液。
(3)抗体药物偶联物的纯化
同实施例1。
(4)抗体药物偶联物分子特性评价
采用上文实验方法部分的“4.4抗体-药物偶联物浓度测定方法-M”中所述操作,测得溶液中抗体药物偶联物的浓度为6.1mg/mL。
采用上文实验方法部分的“5.抗体药物偶联物中每一分子抗体的药物平均连接数(DAR)的测定方法”第5.1.1节中所述操作,测得药物平均连接数(DAR)为3.31(图24)。
采用上文实验方法部分的“6.抗体药物偶联物分子大小异质性测定方法”中所述操作,测得主峰含量为92.16%(图25)。
根据本发明实施例组2抗体药物偶联物的制备中实施例10和11分子排阻色谱分析结果可知,相比8k13-hz35和16k15-hz35,对照抗体IMAB362偶联得到的两份ADC样品IMAB362-M-1和IMAB362-M-2,聚体含量高(检测到单体含量分别为90.13%和92.16%);即,对照抗体IMAB362偶联得到ADC后,理化性质不稳定,易产生聚体,这可能带来各种安全性和有效性方面的问题。另外,疏水分析实验表明,对照抗体IMAB362偶联得到的两份ADC样品,不论加入低当量还原剂(IMAB362-1,抗体摩尔当量的2.2倍)或高当量还原剂(IMAB362-M-2,抗体摩尔当量的8倍),制备得到ADC样品DAR均在3.3左右,且主要制备得到DAR2和DAR4组分,DAR6和DAR8组分很少,药物负载不成正态分布,这可能带来各种药效方面的问题。综合以上两点,对照抗体IMAB362不合适用于制备ADC。
实施例组3 ADC体外细胞活性测试
实施例1 ADC体外细胞活性测试1
使用CLDN18.2-高表达细胞系NCI-N87(NCI-N87 CLDN18.2)作为体外细胞活性实验模型,研究本发明提供的不同抗体药物偶联物对CLDN18.2高表达水平肿瘤细胞的杀伤作用。
实验步骤:根据如上实施例组2中实施例9样品制备方法,将抗体换成Trastuzumab制备得到Trastuzumab-EX(DAR=8)样品。另外采用来自实施例组2中实施例4和5制备的抗体药物偶联物,作为测试药物。将处于对数生长期的上述细胞,分别以每孔5000个细胞的密度接种至96孔细胞培养板中,然后分别加入不同浓度的抗体药物偶联物,每个药物浓度设置2-4个复孔,及空白对照(仅细胞),作用120h(根据细胞生长速度,保证细胞分裂足够次数),测定EC50。
结果见表6。
表6.不同抗体药物偶联物对高表达CLDN18.2的NCI-N87细胞系的细胞杀伤效果
实施例2 ADC体外细胞活性测试2
使用CLDN18.2-高表达细胞系NCI-N87(NCI-N87 CLDN18.2)和CLDN18.2-无表达细胞系NCI-N87(NCI-N87)作为体外细胞活性实验模型,研究本发明提供的不同抗体药物偶联物对CLDN18.2高表达及低表达水平肿瘤细胞的杀伤作用。
实验步骤:测试药物见表7,各个ADC均参照各自实施例制备,其中SAC为Sacituzumab,序列见上文;IL6为TocilizuMab,购自Target Molecule Corp。测试过程同实施例1。
对无CLDN18.2表达的NCI-N87细胞系的细胞杀伤效果见表7和图26。
表7.抗体药物偶联物对无CLDN18.2表达的NCI-N87细胞系的细胞杀伤效果
对CLDN18.2高表达的NCI-N87细胞系的细胞杀伤效果见表8和图27。
表8.抗体药物偶联物对CLDN18.2高表达的NCI-N87细胞系的细胞杀伤效果
样品类型 | 样品名称 | 杀伤作用 | EC50(pM) | DAR |
本发明的ADC-1 | 16k15-hz35-M | + | 63.55 | 4.1 |
本发明的裸抗-1 | 16k15-hz35 | - | - | N/A |
本发明的ADC-2 | 8k13-hz35 | + | 61.58 | 4 |
本发明的裸抗-2 | 8k13-hz35 | - | - | N/A |
阳性对照ADC | SAC-M | + | 64.25 | 4.5 |
阴性对照ADC | IL-6-M | - | - | N/A |
小分子化药 | MMAE | + | 209.7 | N/A |
如结果所示,两种细胞条件下,阳性对照ADC均有杀伤,阴性对照ADC均无杀伤,正负对照结果正常;小分子化药对两种细胞均有杀伤,但量效略有差异,半效杀伤浓度与文献报道一致。同时,两种裸抗分子(16k15-hz35和hz8k13-hz35)对两种细胞均无杀伤,而两种ADC分子(16k15-hz35-M和8k13-hz35-M)对NCI-N87均无杀伤,对NCI-N87 CLDN18.2均有杀伤,这说明NCI-N87 CLDN18.2细胞的杀伤依赖于抗原识别。
实施例3 ADC体外细胞活性测试3
采用本发明的抗体偶联TLR8激动剂得到的ADC并不能直接杀伤肿瘤细胞,而是通过抗体介导肿瘤微环境中的免疫细胞(主要是髓系抗原呈递细胞)吞噬肿瘤细胞后,在免疫细胞溶酶体处释放出TLR8激动剂,TLR8激动剂结合免疫细胞中的TLR8后可激活免疫细胞,提高促炎细胞因子分泌、招募免疫细胞、增强肿瘤新生抗原呈递、刺激肿瘤特异性T细胞生成,最终达到杀灭肿瘤细胞的目的。
使用表达CLDN18.2的NCI-N87细胞系为靶细胞,与人外周血单核细胞(PBMC)共培养,加入一定量的本发明提供的抗体药物偶联物,测定细胞上清中促炎因子的表达情况。
实验步骤:使用新鲜分离的人PBMC或者冻存的PBMC,按照10~30万/孔的数量种入96孔细胞板,37℃,5%CO2培养过夜。第二天按照1万/孔的数量种入NCI-N87 CLDN18.2细胞,然后再加入一定浓度的ADC(实施例组2的实施例1中制备的8k13-hz35-TLR8-1103)、裸抗(8k13-hz35)、阴性对照(NC IgG-TLR8,NC IgG是一种无关人源抗体)或TLR8激动剂(化合物TLR8),共培养24-48h后,取细胞培养上清,ELISA测定上清中促炎细胞因子TNFα浓度。
结果见图28。如结果所示,本发明提供的ADC能明显刺激PBMC分泌促炎因子TNFα,裸抗和阴性对照无明显刺激效果,小分子药物也可以刺激PBMC分泌促炎因子,但强度明显低于本发明提供的ADC。说明将TLR8激动剂偶联在本发明的抗体上以后能通过抗体的靶向作用和Fc效应介导免疫细胞吞噬,将TLR8激动剂递送至靶标部位,激活免疫细胞。
实施例4 ADC体外细胞活性测试4
采用本发明的抗体偶联STING激动剂得到的ADC与表面表达CLDN18.2的肿瘤细胞(NCI-N87 CLDN18.2)孵育后,ADC可以通过靶点的内吞作用进入细胞内部,在细胞内部蛋白酶的作用下断裂linker,释放出STING激动剂。STING激动剂释放以后可以结合并激活STING受体,进而诱导TBK1(TANK binding kinase 1)磷酸化,激活IRF3(Interferon regulatoryfactor 3)信号通路和NF-κB信号通路,IRF3信号通路被激活后将会提高磷酸化TRF3(pIRF3)的表达量。
使用表达CLDN18.2的NCI-N87细胞系为靶细胞,加入一定量的本发明提供的抗体药物偶联物,通过测定pIRF3的表达量可以测定细胞的激活情况。
实验步骤:体外培养NCI-N87 CLDN18.2至对数生长期,按照20万/孔的数量种入24孔细胞板,培养基中不加血清做饥饿处理,37℃,5%CO2培养过夜。第二天加入一定浓度的ADC(实施例组2的实施例2中制备的8k13-hz35-STING-D4-1203)、裸抗(8k13-hz35)、阴性对照(NC IgG-STING,NC IgG是一种无关人源抗体)或STING激动剂(化合物STING),共培养约6h。培养结束后,收集细胞,加入含有蛋白酶抑制剂和磷酸酶抑制剂的细胞裂解液充分裂解细胞,离心收集细胞裂解后上清,通过Western-blotting分析总IRF3和pIRF3,pIRF3与总IRF3表达量的比值即为pIRF3的相对表达量。
结果见图29。如结果所示,本发明的ADC能明显刺激靶细胞中pIRF3表达,裸抗和阴性对照无明显刺激效果,小分子药物也可以刺激pIRF3,但强度明显低于本发明的ADC。说明将STING激动剂偶联在本发明的抗体上以后能通过抗体的靶向作用,将STING激动剂递送至靶标部位,激活STING信号通路。
实施例组4 ADC体内药效测试
实施例1 ADC体内药效测试
细胞:
人胃癌KATO III细胞购自中国科学院细胞库。使KATO III转染人CLDN18.2基因并高表达CLDN18.2蛋白,命名为KATO III/18.2。KATO III/18.2细胞用10cm培养皿培养,培养条件为RPMI 1640培养基中加10%胎牛血清以及青、链霉素,于37℃、含5%CO2空气的培养箱中培养。一周2次传代,当细胞呈指数生长期时,收集细胞,计数,接种。
实验动物:
NOD-Scid小鼠,35日,♀,购自上海灵畅生物科技有限公司。许可证号:SCXK(沪)2018-0003。合格证编号20180003012511。饲养环境:SPF级。
实验步骤:
每只小鼠皮下接种1×107KATO III/18.2细胞,待肿瘤生长至100-150mm3后,根据肿瘤体积将动物分组(D0)。具体给药剂量和给药方案见表9。每周测2次肿瘤体积,称小鼠体重,记录数据。
表9. 8k13-hz35-EX、8k13-hz35-EX2、对照抗体在人胃癌KATO III/18.2荷瘤小鼠的抗肿瘤作用试验设计
注:IV-静脉注射,给药开始于D0。
实验指标:实验指标为考察药物对肿瘤生长的影响,具体指标为T/C(%)或肿瘤生长抑制率%(TGI%)。
肿瘤体积(V)计算公式为V=1/2×a×b2(其中a、b分别表示长、宽);T/C(%)=(T-T0)/(C-C0)×100(其中T、C为实验结束时的肿瘤体积;T0、C0为实验开始时的肿瘤体积);肿瘤生长抑制率%(TGI%)=100-T/C(%);当肿瘤出现消退时,肿瘤生长抑制率%(TGI%)=100-(T-T0)/T0×100;抑瘤率%=(溶剂组肿瘤重量-治疗组肿瘤重量)/溶剂组肿瘤重量×100%。
如果肿瘤比起始体积缩小,即T<T0或C<C0时,即定义为肿瘤部分消退(PR);如果肿瘤完全消失,即定义为肿瘤完全消退(CR)。实验结束、达到实验终点、或肿瘤体积达到1500mm3,CO2麻醉处死动物,随后解剖取瘤并拍照。
结果见表10、表11以及图30至图34。
表10.CLAUDIXIMAB(IMAB362)、8k13-hz35-EX、8k13-hz35-EX2对人胃癌KATO III/18.2小鼠皮下移植瘤的疗效(根据肿瘤体积计算TGI%)
注:P值指与生理盐水组相比。
表11.CLAUDIXIMAB(IMAB362)、8k13-hz35-EX、8k13-hz35-EX2对人胃癌KATO III/18.2小鼠皮下移植瘤的疗效(根据肿瘤重量计算抑瘤率%)
由实验结果可知,CLAUDIXIMAB(IMAB362)(6mg/kg,IV,D0)抑制人胃癌KATO III/18.2小鼠皮下移植瘤的生长,抑瘤率为63%。相比之下,8k13-hz35-EX(DAR8)(3mg/kg,IV,D0)对KATO III/18.2小鼠皮下移植瘤的抑瘤率为98%,有2/6小鼠肿瘤部分消退;8k13-hz35-EX(DAR4)(6mg/kg,IV,D0)对KATO III/18.2小鼠皮下移植瘤的抑瘤率为115%,有3/6小鼠肿瘤部分消退;8k13-hz35-EX2(DAR8)(3、6mg/kg,IV,D0)对KATO III/18.2小鼠皮下移植瘤的抑瘤率分别为74%和122%,分别有3/6和4/6小鼠肿瘤部分消退;8k13-hz35-EX2(DAR4)(3、6mg/kg,IV,D0)对KATO III/18.2小鼠皮下移植瘤的抑瘤率分别为63%和97%,6mg/kg剂量组1/6小鼠肿瘤部分消退(以上药物抑瘤率均根据肿瘤体积计算肿瘤生长抑制率%)。根据肿瘤重量计算抑瘤率得到相似的结果。荷瘤小鼠对以上药物均能很好耐受。
以上对本发明具体实施方式的描述并不限制本发明,本领域技术人员可以根据本发明作出各种改变或变形,只要不脱离本发明的精神,均应属于本发明所附权利要求的范围。
序列表
<110> 江苏迈威康新药研发有限公司
<120> 包含抗CLDN18.2的抗体或其抗原结合片段的抗体药物偶联物及其用途
<130> LC21110001R
<150> CN202110181327.3
<151> 2021-02-09
<160> 49
<170> PatentIn version 3.3
<210> 1
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> 8K13_vh
<400> 1
Gln Val His Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ser
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Asn Tyr
20 25 30
Trp Met Asn Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Gln Ile Tyr Pro Gly Asn Gly Asp Thr Lys Tyr Ser Gly Lys Phe
50 55 60
Asn Ser Lys Asp Thr Leu Thr Ala Asp Lys Ser Ser Asn Thr Ala Tyr
65 70 75 80
Met Gln Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys
85 90 95
Ala Arg Phe Tyr Tyr Gly Asn Val Met Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Ser Val Thr Val Ser Ser
115
<210> 2
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> 8K13_vl
<400> 2
Asp Ile Val Leu Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly
1 5 10 15
Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Thr Leu Leu Asn Gly
20 25 30
Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Ser Gly Gln
35 40 45
Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn
85 90 95
Gly Tyr Ser Tyr Pro Leu Thr Phe Gly Val Gly Thr Lys Leu Glu Leu
100 105 110
Lys
<210> 3
<211> 117
<212> PRT
<213> 人工序列
<220>
<223> 16K15_vh
<400> 3
Glu Val Met Leu Val Glu Ser Gly Gly Gly Leu Val Arg Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Gly Ser Gly Ile Thr Leu Ser Thr Tyr
20 25 30
Ala Met Ser Trp Val Arg Gln Thr Pro Glu Arg Arg Leu Glu Trp Val
35 40 45
Ala Ser Ile Ile Ser Gly Gly Ile Thr Tyr Tyr Leu Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Arg Asn Ile Leu Tyr Leu
65 70 75 80
Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Arg Lys Tyr His Gly Asn Ala Leu Asp Tyr Trp Gly Gln Gly Thr Ser
100 105 110
Val Thr Val Ser Leu
115
<210> 4
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> 16K15_vl
<400> 4
Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Pro Val Thr Ala Gly
1 5 10 15
Glu Thr Val Thr Met Arg Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser
20 25 30
Gly Asn Gln Arg Asn Tyr Leu Thr Trp Tyr Gln Arg Lys Pro Gly Gln
35 40 45
Pro Pro Lys Lys Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Gly Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn
85 90 95
Asn Tyr Phe Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu
100 105 110
Lys
<210> 5
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> 8K13_vh_hz0
<400> 5
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Asn Tyr
20 25 30
Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gln Ile Tyr Pro Gly Asn Gly Asp Thr Lys Tyr Ser Gly Lys Phe
50 55 60
Asn Ser Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Phe Tyr Tyr Gly Asn Val Met Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 6
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> 8K13_vh_hz1
<400> 6
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Asn Tyr
20 25 30
Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gln Ile Tyr Pro Gly Asn Gly Asp Thr Lys Tyr Ser Gly Lys Phe
50 55 60
Asn Ser Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Asn Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Phe Tyr Tyr Gly Asn Val Met Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 7
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> 8K13_vh_hz2
<400> 7
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Asn Tyr
20 25 30
Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gln Ile Tyr Pro Gly Ser Gly Asp Thr Lys Tyr Ser Gly Lys Phe
50 55 60
Gln Ser Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Asn Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Phe Tyr Tyr Gly Asn Val Met Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 8
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> 8K13_vh_hz3
<400> 8
Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Asn Tyr
20 25 30
Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gln Ile Tyr Pro Gly Ser Gly Asp Thr Lys Tyr Ser Gly Lys Phe
50 55 60
Gln Ser Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Phe Tyr Tyr Gly Asn Val Met Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 9
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> 8K13_vl_hz0
<400> 9
Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Thr Leu Leu Asn Gly
20 25 30
Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Asn
85 90 95
Gly Tyr Ser Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
100 105 110
Lys
<210> 10
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> 8K13_vl_hz1
<400> 10
Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Thr Leu Leu Asn Gly
20 25 30
Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn
85 90 95
Gly Tyr Ser Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
100 105 110
Lys
<210> 11
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> 8K13_vl_hz4
<400> 11
Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Thr Leu Leu Ser Gly
20 25 30
Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln
85 90 95
Gly Tyr Ser Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
100 105 110
Lys
<210> 12
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> 8K13_vl_hz5
<400> 12
Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Thr Leu Leu Ser Gly
20 25 30
Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln
85 90 95
Gly Tyr Ser Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
100 105 110
Lys
<210> 13
<211> 117
<212> PRT
<213> 人工序列
<220>
<223> 16k15_vh_hz0
<400> 13
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ile Thr Leu Ser Thr Tyr
20 25 30
Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ser Ile Ile Ser Gly Gly Ile Thr Tyr Tyr Leu Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Lys Tyr His Gly Asn Ala Leu Asp Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser
115
<210> 14
<211> 117
<212> PRT
<213> 人工序列
<220>
<223> 16k15_vh_hz1
<400> 14
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Ile Thr Leu Ser Thr Tyr
20 25 30
Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ser Ile Ile Ser Gly Gly Ile Thr Tyr Tyr Leu Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Lys Tyr His Gly Asn Ala Leu Asp Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser
115
<210> 15
<211> 117
<212> PRT
<213> 人工序列
<220>
<223> 16k15_vh_hz2
<400> 15
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Ile Thr Leu Ser Thr Tyr
20 25 30
Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ser Ile Ile Ser Gly Gly Ile Thr Tyr Tyr Leu Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr Leu
65 70 75 80
Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Lys Tyr His Gly Asn Ala Leu Asp Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser
115
<210> 16
<211> 117
<212> PRT
<213> 人工序列
<220>
<223> 16k15_vh_hz3
<400> 16
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Ile Thr Leu Ser Thr Tyr
20 25 30
Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ser Ile Ile Ser Gly Gly Ile Thr Tyr Tyr Leu Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Lys Tyr His Gly Asn Ala Leu Asp Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser
115
<210> 17
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> 16K15_vl_hz0
<400> 17
Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser
20 25 30
Gly Asn Gln Arg Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Asn
85 90 95
Asn Tyr Phe Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
100 105 110
Lys
<210> 18
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> 16K15_vl_hz1
<400> 18
Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser
20 25 30
Gly Asn Gln Arg Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Lys Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn
85 90 95
Asn Tyr Phe Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
100 105 110
Lys
<210> 19
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> 16K15_vl_hz2
<400> 19
Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Leu Ser Ser
20 25 30
Gly Asn Gln Arg Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Lys Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn
85 90 95
Asn Tyr Phe Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
100 105 110
Lys
<210> 20
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> 16K15_vl_hz3
<400> 20
Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Leu Ser Ser
20 25 30
Gly Asn Gln Arg Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Lys Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Ser
85 90 95
Asn Tyr Phe Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
100 105 110
Lys
<210> 21
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> 16K15_vl_hz4
<400> 21
Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Leu Ser Ser
20 25 30
Gly Asn Gln Arg Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Lys Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln
85 90 95
Asn Tyr Phe Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
100 105 110
Lys
<210> 22
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> 16k15_vl_hz5
<400> 22
Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Leu Ser Ser
20 25 30
Gly Asn Gln Arg Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Lys Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln
85 90 95
Asn Tyr Phe Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
100 105 110
Lys
<210> 23
<211> 10
<212> PRT
<213> 人工序列
<220>
<223> CDR-H1
<400> 23
Gly Tyr Ala Phe Ser Asn Tyr Trp Met Asn
1 5 10
<210> 24
<211> 18
<212> PRT
<213> 人工序列
<220>
<223> CDR-H2
<400> 24
Gly Gln Ile Tyr Pro Gly Asn Gly Asp Thr Lys Tyr Ser Gly Lys Phe
1 5 10 15
Asn Ser
<210> 25
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> CDR-H3
<400> 25
Phe Tyr Tyr Gly Asn Val Met Asp Tyr
1 5
<210> 26
<211> 18
<212> PRT
<213> 人工序列
<220>
<223> CDR-H2
<400> 26
Gly Gln Ile Tyr Pro Gly Ser Gly Asp Thr Lys Tyr Ser Gly Lys Phe
1 5 10 15
Gln Ser
<210> 27
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> CDR-H1
<400> 27
Gly Tyr Ala Phe Ser Asn Tyr
1 5
<210> 28
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> CDR-H2
<400> 28
Tyr Pro Gly Ser Gly Asp
1 5
<210> 29
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> CDR-L1
<400> 29
Lys Ser Ser Gln Thr Leu Leu Asn Gly Gly Asn Gln Lys Asn Tyr Leu
1 5 10 15
Thr
<210> 30
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> CDR-L2
<400> 30
Trp Ala Ser Thr Arg Glu Ser
1 5
<210> 31
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> CDR-L3
<400> 31
Gln Asn Gly Tyr Ser Tyr Pro Leu Thr
1 5
<210> 32
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> CDR-L1
<400> 32
Lys Ser Ser Gln Thr Leu Leu Ser Gly Gly Asn Gln Lys Asn Tyr Leu
1 5 10 15
Thr
<210> 33
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> CDR-L3
<400> 33
Gln Gln Gly Tyr Ser Tyr Pro Leu Thr
1 5
<210> 34
<211> 10
<212> PRT
<213> 人工序列
<220>
<223> CDR-H1
<400> 34
Gly Ile Thr Leu Ser Thr Tyr Ala Met Ser
1 5 10
<210> 35
<211> 16
<212> PRT
<213> 人工序列
<220>
<223> CDR-H2
<400> 35
Ser Ile Ile Ser Gly Gly Ile Thr Tyr Tyr Leu Asp Ser Val Lys Gly
1 5 10 15
<210> 36
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> CDR-H3
<400> 36
Lys Tyr His Gly Asn Ala Leu Asp Tyr
1 5
<210> 37
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> CDR-H1
<400> 37
Gly Ile Thr Leu Ser Thr Tyr
1 5
<210> 38
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> CDR-H2
<400> 38
Ile Ser Gly Gly Ile Thr
1 5
<210> 39
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> CDR-L1
<400> 39
Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asn Gln Arg Asn Tyr Leu
1 5 10 15
Thr
<210> 40
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> CDR-L3
<400> 40
Gln Asn Asn Tyr Phe Tyr Pro Leu Thr
1 5
<210> 41
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> CDR-L1
<400> 41
Lys Ser Ser Gln Ser Leu Leu Ser Ser Gly Asn Gln Arg Asn Tyr Leu
1 5 10 15
Thr
<210> 42
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> CDR-L3
<400> 42
Gln Ser Asn Tyr Phe Tyr Pro Leu Thr
1 5
<210> 43
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> CDR-L3
<400> 43
Gln Gln Asn Tyr Phe Tyr Pro Leu Thr
1 5
<210> 44
<211> 448
<212> PRT
<213> 人工序列
<220>
<223> IMAB362,重链
<400> 44
Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Ala
1 5 10 15
Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Trp Ile Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Asn Ile Tyr Pro Ser Asp Ser Tyr Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Asp Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Pro Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Thr Arg Ser Trp Arg Gly Asn Ser Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
130 135 140
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn
145 150 155 160
Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
165 170 175
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser
180 185 190
Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr
210 215 220
His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser
225 230 235 240
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
245 250 255
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
275 280 285
Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
290 295 300
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
305 310 315 320
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
325 330 335
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350
Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys
355 360 365
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
405 410 415
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
420 425 430
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 45
<211> 220
<212> PRT
<213> 人工序列
<220>
<223> IMAB362,轻链
<400> 45
Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly
1 5 10 15
Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser
20 25 30
Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn
85 90 95
Asp Tyr Ser Tyr Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile
100 105 110
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
115 120 125
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn
130 135 140
Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu
145 150 155 160
Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
165 170 175
Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr
180 185 190
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
195 200 205
Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215 220
<210> 46
<211> 451
<212> PRT
<213> 人工序列
<220>
<223> Sacituzumab,重链
<400> 46
Gln Val Gln Leu Gln Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr
20 25 30
Gly Met Asn Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Lys Trp Met
35 40 45
Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Thr Asp Asp Phe
50 55 60
Lys Gly Arg Phe Ala Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr
65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Gly Gly Phe Gly Ser Ser Tyr Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Ser Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
130 135 140
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
145 150 155 160
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
180 185 190
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys
210 215 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
225 230 235 240
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
275 280 285
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
305 310 315 320
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
340 345 350
Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
355 360 365
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
385 390 395 400
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
420 425 430
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445
Pro Gly Lys
450
<210> 47
<211> 214
<212> PRT
<213> 人工序列
<220>
<223> Sacituzumab,轻链
<400> 47
Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Ile Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln His Tyr Ile Thr Pro Leu
85 90 95
Thr Phe Gly Ala Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 48
<211> 330
<212> PRT
<213> 人工序列
<220>
<223> 重链,恒定区
<400> 48
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
1 5 10 15
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
195 200 205
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
210 215 220
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu
225 230 235 240
Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
245 250 255
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
260 265 270
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
275 280 285
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
290 295 300
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
305 310 315 320
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330
<210> 49
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> 轻链,恒定区
<400> 49
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
1 5 10 15
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
20 25 30
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
35 40 45
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
50 55 60
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
65 70 75 80
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
85 90 95
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
100 105
Claims (11)
1.一种抗体药物偶联物,其包含与药物共价连接的抗CLDN18.2的抗体或其片段,其中所述抗CLDN18.2的抗体或其片段包含重链和轻链,所述重链和轻链分别包含如下所示的重链互补决定区1至3(CDR-H1、CDR-H2和CDR-H3)和轻链互补决定区1至3(CDR-L1、CDR-L2和CDR-L3):
(i)氨基酸序列分别如SEQ ID NO:23、SEQ ID NO:26和SEQ ID NO:25所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:32、SEQ ID NO:30和SEQ ID NO:33所示的CDR-L1、CDR-L2和CDR-L3;
(ii)氨基酸序列分别如SEQ ID NO:27、SEQ ID NO:28和SEQ ID NO:25所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:32、SEQ ID NO:30和SEQ ID NO:33所示的CDR-L1、CDR-L2和CDR-L3;
(iii)氨基酸序列分别如SEQ ID NO:34、SEQ ID NO:35和SEQ ID NO:36所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:41、SEQ ID NO:30和SEQ ID NO:40所示的CDR-L1、CDR-L2和CDR-L3;
(iv)氨基酸序列分别如SEQ ID NO:37、SEQ ID NO:38和SEQ ID NO:36所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:41、SEQ ID NO:30和SEQ ID NO:43所示的CDR-L1、CDR-L2和CDR-L3;或
(v)氨基酸序列分别如SEQ ID NO:34、SEQ ID NO:35和SEQ ID NO:36所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:41、SEQ ID NO:30和SEQ ID NO:43所示的CDR-L1、CDR-L2和CDR-L3。
2.根据权利要求1所述的抗体药物偶联物,其特征在于,所述抗CLDN18.2的抗体或其片段的重链包含重链可变区(VH),所述重链可变区(VH)包含SEQ ID NO:7或SEQ ID NO:8所示的氨基酸序列或其变体;和/或,所述抗CLDN18.2的抗体或其片段的轻链包含轻链可变区(VL),所述轻链可变区(VL)包含:选自SEQ ID NO:11或SEQ ID NO:12所示的氨基酸序列或其变体;
或者,优选地,所述抗CLDN18.2的抗体或其片段的重链包含重链可变区(VH),所述重链可变区(VH)包含SEQ ID NO:15或SEQ ID NO:16所示的氨基酸序列或其变体;和/或,所述抗CLDN18.2的抗体或其片段的轻链包含轻链可变区(VL),所述轻链可变区(VL)包含选自SEQID NO:19或SEQ ID NO:22所示的氨基酸序列或其变体;
更优选地,所述抗CLDN18.2的抗体或其片段的重链和轻链分别包含如下所示的重链可变区(VH)和轻链可变区(VL):
(i)如SEQ ID NO:7所示的氨基酸序列或其变体;和,如SEQ ID NO:11所示的氨基酸序列或其变体;
(ii)如SEQ ID NO:8所示的氨基酸序列或其变体;和,如SEQ ID NO:12所示的氨基酸序列或其变体;
(iii)如SEQ ID NO:15所示的氨基酸序列或其变体;和,如SEQ ID NO:19所示的氨基酸序列或其变体;
(iv)如SEQ ID NO:16所示的氨基酸序列或其变体;和,如SEQ ID NO:22所示的氨基酸序列或其变体。
3.根据权利要求1或2所述的抗体药物偶联物,其特征在于,所述抗CLDN18.2的抗体或其片段为针对CLDN18.2的单克隆抗体、单链抗体、双功能抗体、单域抗体、纳米抗体、完全或部分人源化的抗体或者嵌合抗体等任意形式;或者,所述抗CLDN18.2的抗体或其片段为针对CLDN18.2的半抗体或半抗体的抗原结合片段;
任选地,所述抗CLDN18.2的抗体或其片段为scFv、BsFv、dsFv、(dsFv)2、Fab、Fab'、F(ab')2或Fv;
优选地,所述抗体或其片段还包含人或鼠的恒定区,优选包含人或鼠的轻链恒定区(CL)和/或重链恒定区(CH);
更优选地,所述抗体或其抗原结合片段包含选自IgG、IgA、IgM、IgD或IgE的重链恒定区和/或κ或λ型轻链恒定区。
4.根据权利要求1至3中任一项所述的抗体药物偶联物,其特征在于,所述抗体药物偶联物具有分子式Ab-[L-D]n,其中Ab表示抗CLDN18.2的抗体或其片段,L表示接头,D表示药物,n表示相对于每一分子Ab的药物平均连接数。
5.根据权利要求1至4中任一项所述的抗体药物偶联物,其特征在于,所述药物D为细胞毒性类小分子药物和/或免疫治疗剂类小分子药物;
优选地,所述细胞毒性类小分子药物为微管蛋白抑制剂(如澳瑞他汀类化合物)和/或拓扑异构酶I抑制剂(如喜树碱类化合物);
优选地,所述免疫治疗剂类小分子药物为STING激动剂(如环二核苷酸类化合物)和/或TLR7/8激动剂(如咪唑并喹啉类化合物和/或苯并氮卓类化合物);
优选地,所述接头L为半胱氨酸偶联接头或赖氨酸偶联接头;
优选地,所述接头L为可切割或不可切割的;
优选地,n为1至12、优选1至8、更优选3至8。
6.权利要求1至5中任一项所述的抗体药物偶联物的制备方法,所述方法包括以下步骤:
(1)用抗体与还原试剂在缓冲液中反应,得到经还原后的抗体;
(2)用药物接头与步骤(1)得到的经还原后的抗体在缓冲液与有机溶剂的混合液中进行交联,得到抗体药物偶联物。
7.一种抗CLDN18.2的抗体或其片段,所述抗CLDN18.2的抗体或其片段包含重链和轻链,所述重链和轻链分别包含如下所示的重链互补决定区1至3(CDR-H1、CDR-H2和CDR-H3)和轻链互补决定区1至3(CDR-L1、CDR-L2和CDR-L3):
(i)氨基酸序列分别如SEQ ID NO:23、SEQ ID NO:26和SEQ ID NO:25所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:32、SEQ ID NO:30和SEQ ID NO:33所示的CDR-L1、CDR-L2和CDR-L3;
(ii)氨基酸序列分别如SEQ ID NO:27、SEQ ID NO:28和SEQ ID NO:25所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:32、SEQ ID NO:30和SEQ ID NO:33所示的CDR-L1、CDR-L2和CDR-L3;
(iii)氨基酸序列分别如SEQ ID NO:34、SEQ ID NO:35和SEQ ID NO:36所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:41、SEQ ID NO:30和SEQ ID NO:40所示的CDR-L1、CDR-L2和CDR-L3;
(iv)氨基酸序列分别如SEQ ID NO:37、SEQ ID NO:38和SEQ ID NO:36所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:41、SEQ ID NO:30和SEQ ID NO:43所示的CDR-L1、CDR-L2和CDR-L3;或
(v)氨基酸序列分别如SEQ ID NO:34、SEQ ID NO:35和SEQ ID NO:36所示的CDR-H1、CDR-H2和CDR-H3;和,氨基酸序列分别如SEQ ID NO:41、SEQ ID NO:30和SEQ ID NO:43所示的CDR-L1、CDR-L2和CDR-L3。
8.根据权利要求7所述的抗CLDN18.2的抗体或其片段,其特征在于,所述抗CLDN18.2的抗体或其片段的重链包含重链可变区(VH),轻链包含轻链可变区(VL),所述重链可变区(VH)和轻链可变区(VL)分别包含SEQ ID NO:8所示的氨基酸序列或其变体和SEQ ID NO:12所示的氨基酸序列或其变体;或者,所述重链可变区(VH)和轻链可变区(VL)分别包含SEQID NO:16所示的氨基酸序列或其变体和SEQ ID NO:22所示的氨基酸序列或其变体。
9.一种药物组合物,其包含权利要求1至5中任一项所述的抗体药物偶联物或者权利要求7至8中任一项所述的抗CLDN18.2的抗体或其片段。
10.一种药物组合物,其包含抗体药物偶联物混合物和药学上可接受的载体,所述抗体药物偶联物混合物包含多种如权利要求1至5中任一项所述的抗体药物偶联物;
任选地,所述抗体药物偶联物混合物的平均n值为2至8;
任选地,所述抗体药物偶联物混合物包含多种所述抗体药物偶联物,每种所述抗体药物偶联物的n值为1至12、优选1至8、更优选3至8。
11.权利要求1至5中任一项所述的抗体药物偶联物、权利要求7至8中任一项所述的抗CLDN18.2的抗体或其片段或者权利要求9至10中任一项所述的药物组合物在制备药物中的用途,所述药物用于治疗与CLDN18.2相关的疾病,优选为肿瘤,更优选为癌症。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN2021101813273 | 2021-02-09 | ||
CN202110181327 | 2021-02-09 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN114904015A true CN114904015A (zh) | 2022-08-16 |
Family
ID=82763528
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210133787.3A Pending CN114904015A (zh) | 2021-02-09 | 2022-02-09 | 包含抗cldn18.2的抗体或其抗原结合片段的抗体药物偶联物及其用途 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN114904015A (zh) |
WO (1) | WO2022171134A1 (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN117327182A (zh) * | 2023-09-19 | 2024-01-02 | 上海交通大学医学院附属仁济医院 | Cldn18.2单域抗体探针的制备方法及应用 |
WO2024114667A1 (zh) * | 2022-11-30 | 2024-06-06 | 正大天晴药业集团股份有限公司 | 抗cldn18.2抗体药物偶联物及其药物组合物和用途 |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016165762A1 (en) * | 2015-04-15 | 2016-10-20 | Ganymed Pharmaceuticals Ag | Drug conjugates comprising antibodies against claudin 18.2 |
CN111110862A (zh) * | 2018-11-01 | 2020-05-08 | 上海健信生物医药科技有限公司 | 抗cldn18.2抗体的药物偶联体及其制备方法和用途 |
CN112574307B (zh) * | 2019-09-29 | 2023-11-28 | 迈威(上海)生物科技股份有限公司 | 抗人Claudin18.2抗体及其应用 |
-
2022
- 2022-02-09 WO PCT/CN2022/075689 patent/WO2022171134A1/zh active Application Filing
- 2022-02-09 CN CN202210133787.3A patent/CN114904015A/zh active Pending
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2024114667A1 (zh) * | 2022-11-30 | 2024-06-06 | 正大天晴药业集团股份有限公司 | 抗cldn18.2抗体药物偶联物及其药物组合物和用途 |
CN117327182A (zh) * | 2023-09-19 | 2024-01-02 | 上海交通大学医学院附属仁济医院 | Cldn18.2单域抗体探针的制备方法及应用 |
CN117327182B (zh) * | 2023-09-19 | 2024-06-04 | 上海交通大学医学院附属仁济医院 | Cldn18.2单域抗体探针的制备方法及应用 |
Also Published As
Publication number | Publication date |
---|---|
WO2022171134A1 (zh) | 2022-08-18 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN112543771B (zh) | 抗b7h3抗体-依喜替康类似物偶联物及其医药用途 | |
JP7118117B2 (ja) | 抗体-薬物コンジュゲートの選択的製造方法 | |
CN112351797B (zh) | 抗b7-h4抗体-药物偶联物及其医药用途 | |
CN109963591B (zh) | B7h3抗体-药物偶联物及其医药用途 | |
EP3743081A1 (en) | Antibodies specific to delta 1 chain of t cell receptor | |
TWI744261B (zh) | 抗5t4抗體和抗體-藥物共軛體 | |
CA3075371A1 (en) | Potentiating the effect of atp release | |
US20220356246A1 (en) | Anti-ROR1 antibodies and preparation method and uses thereof | |
WO2021248048A9 (en) | Antibody-drug conjugates containing an anti-mesothelin antibody and uses thereof | |
CN115605510A (zh) | B7h3抗体-依喜替康类似物偶联物及其医药用途 | |
CN112955548B (zh) | 叶酸受体α特异性抗体 | |
CN114904015A (zh) | 包含抗cldn18.2的抗体或其抗原结合片段的抗体药物偶联物及其用途 | |
WO2021190480A1 (zh) | 抗体-药物偶联物及其医药用途 | |
KR20220075393A (ko) | Bcma를 표적으로 하는 항체, 이중 특이성 항체 및 이의 용도 | |
CN114269389B (zh) | 靶向紧密连接蛋白18.2的抗体药物偶联物 | |
CN112390885A (zh) | 一种trop2抗体及其制备方法、其偶联物和应用 | |
CN116261595A (zh) | 抗vegf-抗pd-l1双特异性抗体、其药物组合物及用途 | |
CN112585168B (zh) | 抗bcma抗体、其抗原结合片段及其医药用途 | |
WO2021190564A1 (zh) | 抗体药物偶联物及其医药用途 | |
WO2022228563A1 (zh) | 靶向Nectin-4的抗体药物偶联物及其制备方法和用途 | |
CN117460540A (zh) | 艾日布林衍生物的药物偶联物 | |
CN112500491A (zh) | 一种特异性中和辅助性T细胞TGF-β信号的双特异性抗体、其药物组合及其用途 | |
JP2021521872A (ja) | C−metに特異的に結合する抗体及びその用途 | |
WO2024131835A1 (en) | Ligand-cytotoxicity drug conjugates and pharmaceutical uses thereof | |
RU2785664C2 (ru) | Конъюгат антитело к b7h3-аналог экзатекана и его применение в медицине |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |