CN116096747A - 免疫原性降低型低分子抗体及其制造法 - Google Patents
免疫原性降低型低分子抗体及其制造法 Download PDFInfo
- Publication number
- CN116096747A CN116096747A CN202180056535.9A CN202180056535A CN116096747A CN 116096747 A CN116096747 A CN 116096747A CN 202180056535 A CN202180056535 A CN 202180056535A CN 116096747 A CN116096747 A CN 116096747A
- Authority
- CN
- China
- Prior art keywords
- ser
- gly
- ala
- val
- thr
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000000034 method Methods 0.000 title claims abstract description 82
- 230000005847 immunogenicity Effects 0.000 title description 20
- 230000002829 reductive effect Effects 0.000 title description 11
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims abstract description 75
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 56
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 55
- 150000008574 D-amino acids Chemical class 0.000 claims abstract description 53
- 239000004471 Glycine Substances 0.000 claims abstract description 37
- 238000012216 screening Methods 0.000 claims abstract description 21
- 238000004519 manufacturing process Methods 0.000 claims abstract description 17
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 122
- 150000001413 amino acids Chemical group 0.000 claims description 95
- 108010003723 Single-Domain Antibodies Proteins 0.000 claims description 84
- 238000003786 synthesis reaction Methods 0.000 claims description 81
- 230000015572 biosynthetic process Effects 0.000 claims description 73
- 239000012634 fragment Substances 0.000 claims description 41
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 20
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 claims description 17
- 229920001184 polypeptide Polymers 0.000 claims description 15
- 125000003295 alanine group Chemical group N[C@@H](C)C(=O)* 0.000 claims description 12
- 238000010532 solid phase synthesis reaction Methods 0.000 claims description 9
- 230000002194 synthesizing effect Effects 0.000 claims description 7
- ZRALSGWEFCBTJO-UHFFFAOYSA-N Guanidine Chemical compound NC(N)=N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 description 54
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 49
- 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 48
- 235000001014 amino acid Nutrition 0.000 description 42
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 42
- 108010067216 glycyl-glycyl-glycine Proteins 0.000 description 41
- 108010090333 leucyl-lysyl-proline Proteins 0.000 description 40
- 238000005259 measurement Methods 0.000 description 40
- 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 39
- 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 39
- 108010008355 arginyl-glutamine Proteins 0.000 description 39
- 238000004128 high performance liquid chromatography Methods 0.000 description 39
- 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 38
- 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 38
- 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 37
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 37
- 239000011347 resin Substances 0.000 description 33
- 229920005989 resin Polymers 0.000 description 33
- 239000012043 crude product Substances 0.000 description 30
- 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 29
- ZTLGVASZOIKNIX-DCAQKATOSA-N Leu-Gln-Glu Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N ZTLGVASZOIKNIX-DCAQKATOSA-N 0.000 description 29
- 108010020755 prolyl-glycyl-glycine Proteins 0.000 description 29
- 239000000243 solution Substances 0.000 description 28
- 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 27
- AXZGZMGRBDQTEY-SRVKXCTJSA-N Leu-Gln-Met Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCSC)C(O)=O AXZGZMGRBDQTEY-SRVKXCTJSA-N 0.000 description 27
- CHJJGSNFBQVOTG-UHFFFAOYSA-N N-methyl-guanidine Natural products CNC(N)=N CHJJGSNFBQVOTG-UHFFFAOYSA-N 0.000 description 27
- SWSQBOPZIKWTGO-UHFFFAOYSA-N dimethylaminoamidine Natural products CN(C)C(N)=N SWSQBOPZIKWTGO-UHFFFAOYSA-N 0.000 description 27
- 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 26
- 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 26
- AJHCSUXXECOXOY-UHFFFAOYSA-N N-glycyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)CN)C(O)=O)=CNC2=C1 AJHCSUXXECOXOY-UHFFFAOYSA-N 0.000 description 26
- 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 25
- 239000008363 phosphate buffer Substances 0.000 description 25
- QEDMOZUJTGEIBF-FXQIFTODSA-N Ser-Arg-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O QEDMOZUJTGEIBF-FXQIFTODSA-N 0.000 description 24
- 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 23
- 102000036639 antigens Human genes 0.000 description 23
- 108091007433 antigens Proteins 0.000 description 23
- 238000000746 purification Methods 0.000 description 23
- 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 22
- 241001416177 Vicugna pacos Species 0.000 description 22
- 239000000427 antigen Substances 0.000 description 22
- RLSSMJSEOOYNOY-UHFFFAOYSA-N m-cresol Chemical compound CC1=CC=CC(O)=C1 RLSSMJSEOOYNOY-UHFFFAOYSA-N 0.000 description 22
- FGWUALWGCZJQDJ-URLPEUOOSA-N Phe-Thr-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O FGWUALWGCZJQDJ-URLPEUOOSA-N 0.000 description 21
- SLOYNOMYOAOUCX-BVSLBCMMSA-N Trp-Phe-Arg Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O SLOYNOMYOAOUCX-BVSLBCMMSA-N 0.000 description 21
- 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 21
- 230000027455 binding Effects 0.000 description 21
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 21
- CNGOEHJCLVCJHN-SRVKXCTJSA-N Lys-Pro-Glu Chemical compound NCCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O CNGOEHJCLVCJHN-SRVKXCTJSA-N 0.000 description 20
- 230000008569 process Effects 0.000 description 20
- 241000282828 Camelus bactrianus Species 0.000 description 19
- UIDJDMVRDUANDL-BVSLBCMMSA-N Trp-Tyr-Arg Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O UIDJDMVRDUANDL-BVSLBCMMSA-N 0.000 description 19
- 238000005406 washing Methods 0.000 description 19
- 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 18
- 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 18
- 241000880493 Leptailurus serval Species 0.000 description 18
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 18
- 102100035360 Cerebellar degeneration-related antigen 1 Human genes 0.000 description 17
- 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 17
- 235000018102 proteins Nutrition 0.000 description 17
- SLKLLQWZQHXYSV-CIUDSAMLSA-N Asn-Ala-Lys Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O SLKLLQWZQHXYSV-CIUDSAMLSA-N 0.000 description 16
- 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 16
- PZBFGYYEXUXCOF-UHFFFAOYSA-N TCEP Chemical compound OC(=O)CCP(CCC(O)=O)CCC(O)=O PZBFGYYEXUXCOF-UHFFFAOYSA-N 0.000 description 16
- 238000006243 chemical reaction Methods 0.000 description 16
- 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 15
- HHWQMFIGMMOVFK-WDSKDSINSA-N Gln-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(N)=O HHWQMFIGMMOVFK-WDSKDSINSA-N 0.000 description 15
- DKDHTRVDOUZZTP-IFFSRLJSSA-N Thr-Gln-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)[C@@H](C)O)C(O)=O DKDHTRVDOUZZTP-IFFSRLJSSA-N 0.000 description 15
- BPGDJSUFQKWUBK-KJEVXHAQSA-N Thr-Val-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 BPGDJSUFQKWUBK-KJEVXHAQSA-N 0.000 description 15
- 108010076324 alanyl-glycyl-glycine Proteins 0.000 description 15
- 244000303258 Annona diversifolia Species 0.000 description 14
- 235000002198 Annona diversifolia Nutrition 0.000 description 14
- 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 14
- MNYNCKZAEIAONY-XGEHTFHBSA-N Thr-Val-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O MNYNCKZAEIAONY-XGEHTFHBSA-N 0.000 description 14
- 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 14
- 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 14
- 108010059459 arginyl-threonyl-phenylalanine Proteins 0.000 description 14
- BCADFFUQHIMQAA-KKHAAJSZSA-N Asn-Thr-Val 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)C)C(O)=O BCADFFUQHIMQAA-KKHAAJSZSA-N 0.000 description 13
- BUDNAJYVCUHLSV-ZLUOBGJFSA-N Ala-Asp-Ser Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O BUDNAJYVCUHLSV-ZLUOBGJFSA-N 0.000 description 12
- NKBQZKVMKJJDLX-SRVKXCTJSA-N Arg-Glu-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O NKBQZKVMKJJDLX-SRVKXCTJSA-N 0.000 description 12
- 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 12
- HNAUFGBKJLTWQE-IFFSRLJSSA-N Gln-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CCC(=O)N)N)O HNAUFGBKJLTWQE-IFFSRLJSSA-N 0.000 description 12
- MGDFPGCFVJFITQ-CIUDSAMLSA-N Pro-Glu-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O MGDFPGCFVJFITQ-CIUDSAMLSA-N 0.000 description 12
- 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 12
- JXWGBRRVTRAZQA-ULQDDVLXSA-N Val-Tyr-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](C(C)C)N JXWGBRRVTRAZQA-ULQDDVLXSA-N 0.000 description 12
- 239000000872 buffer Substances 0.000 description 12
- 108010038745 tryptophylglycine Proteins 0.000 description 12
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 11
- 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 11
- SVGAWGVHFIYAEE-JSGCOSHPSA-N Trp-Gly-Gln Chemical compound C1=CC=C2C(C[C@H](N)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(O)=O)=CNC2=C1 SVGAWGVHFIYAEE-JSGCOSHPSA-N 0.000 description 11
- 108010047495 alanylglycine Proteins 0.000 description 11
- 238000004458 analytical method Methods 0.000 description 11
- HNKJADCVZUBCPG-UHFFFAOYSA-N thioanisole Chemical compound CSC1=CC=CC=C1 HNKJADCVZUBCPG-UHFFFAOYSA-N 0.000 description 11
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 10
- DTPOVRRYXPJJAZ-FJXKBIBVSA-N Gly-Arg-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCN=C(N)N DTPOVRRYXPJJAZ-FJXKBIBVSA-N 0.000 description 10
- 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 10
- 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 10
- 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 10
- 239000000611 antibody drug conjugate Substances 0.000 description 10
- 229940049595 antibody-drug conjugate Drugs 0.000 description 10
- 238000010511 deprotection reaction Methods 0.000 description 10
- 108010078144 glutaminyl-glycine Proteins 0.000 description 10
- 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 9
- FEZJJKXNPSEYEV-CIUDSAMLSA-N Arg-Gln-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O FEZJJKXNPSEYEV-CIUDSAMLSA-N 0.000 description 9
- TVYMKYUSZSVOAG-ZLUOBGJFSA-N Cys-Ala-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O TVYMKYUSZSVOAG-ZLUOBGJFSA-N 0.000 description 9
- WNZOCXUOGVYYBJ-CDMKHQONSA-N Gly-Phe-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)CN)O WNZOCXUOGVYYBJ-CDMKHQONSA-N 0.000 description 9
- LCRDMSSAKLTKBU-ZDLURKLDSA-N Gly-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)CN LCRDMSSAKLTKBU-ZDLURKLDSA-N 0.000 description 9
- ZLCLYFGMKFCDCN-XPUUQOCRSA-N Gly-Ser-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CO)NC(=O)CN)C(O)=O ZLCLYFGMKFCDCN-XPUUQOCRSA-N 0.000 description 9
- XOGTZOOQQBDUSI-UHFFFAOYSA-M Mesna Chemical compound [Na+].[O-]S(=O)(=O)CCS XOGTZOOQQBDUSI-UHFFFAOYSA-M 0.000 description 9
- 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 9
- AEGUWTFAQQWVLC-BQBZGAKWSA-N Ser-Gly-Arg Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O AEGUWTFAQQWVLC-BQBZGAKWSA-N 0.000 description 9
- 238000001142 circular dichroism spectrum Methods 0.000 description 9
- 238000003776 cleavage reaction Methods 0.000 description 9
- 238000001914 filtration Methods 0.000 description 9
- 229960004635 mesna Drugs 0.000 description 9
- CMWYAOXYQATXSI-UHFFFAOYSA-N n,n-dimethylformamide;piperidine Chemical compound CN(C)C=O.C1CCNCC1 CMWYAOXYQATXSI-UHFFFAOYSA-N 0.000 description 9
- 239000002244 precipitate Substances 0.000 description 9
- 230000007017 scission Effects 0.000 description 9
- 238000010561 standard procedure Methods 0.000 description 9
- 108010073969 valyllysine Proteins 0.000 description 9
- 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 8
- SKTGPBFTMNLIHQ-KKUMJFAQSA-N Arg-Glu-Phe Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O SKTGPBFTMNLIHQ-KKUMJFAQSA-N 0.000 description 8
- 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 8
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 8
- 235000018417 cysteine Nutrition 0.000 description 8
- 239000003814 drug Substances 0.000 description 8
- 108010084389 glycyltryptophan Proteins 0.000 description 8
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 8
- 230000004048 modification Effects 0.000 description 8
- 238000012986 modification Methods 0.000 description 8
- 108010009962 valyltyrosine Proteins 0.000 description 8
- VNYMOTCMNHJGTG-JBDRJPRFSA-N Ala-Ile-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O VNYMOTCMNHJGTG-JBDRJPRFSA-N 0.000 description 7
- 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 7
- JQFJNGVSGOUQDH-XIRDDKMYSA-N Arg-Glu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCN=C(N)N)N)C(O)=O)=CNC2=C1 JQFJNGVSGOUQDH-XIRDDKMYSA-N 0.000 description 7
- SBCYJMOOHUDWDA-NUMRIWBASA-N Glu-Asp-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SBCYJMOOHUDWDA-NUMRIWBASA-N 0.000 description 7
- 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 7
- 150000008575 L-amino acids Chemical class 0.000 description 7
- RTIRBWJPYJYTLO-MELADBBJSA-N Leu-Lys-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@@H]1C(=O)O)N RTIRBWJPYJYTLO-MELADBBJSA-N 0.000 description 7
- 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 7
- HAAQQNHQZBOWFO-LURJTMIESA-N Pro-Gly-Gly Chemical compound OC(=O)CNC(=O)CNC(=O)[C@@H]1CCCN1 HAAQQNHQZBOWFO-LURJTMIESA-N 0.000 description 7
- 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 7
- 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 7
- 238000002835 absorbance Methods 0.000 description 7
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 7
- -1 fluidity promoters Substances 0.000 description 7
- 108010000434 glycyl-alanyl-leucine Proteins 0.000 description 7
- 150000007970 thio esters Chemical class 0.000 description 7
- 108010003137 tyrosyltyrosine Proteins 0.000 description 7
- NXLNNXIXOYSCMB-UHFFFAOYSA-N (4-nitrophenyl) carbonochloridate Chemical compound [O-][N+](=O)C1=CC=C(OC(Cl)=O)C=C1 NXLNNXIXOYSCMB-UHFFFAOYSA-N 0.000 description 6
- UZSQXCMNUPKLCC-FJXKBIBVSA-N Arg-Thr-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O UZSQXCMNUPKLCC-FJXKBIBVSA-N 0.000 description 6
- KABHAOSDMIYXTR-GUBZILKMSA-N Cys-Glu-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CS)N KABHAOSDMIYXTR-GUBZILKMSA-N 0.000 description 6
- 101001065501 Escherichia phage MS2 Lysis protein Proteins 0.000 description 6
- VEPBEGNDJYANCF-QWRGUYRKSA-N Gly-Lys-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCCN VEPBEGNDJYANCF-QWRGUYRKSA-N 0.000 description 6
- 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 6
- XNMYNGDKJNOKHH-BZSNNMDCSA-N Phe-Ser-Tyr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O XNMYNGDKJNOKHH-BZSNNMDCSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- GRIUMVXCJDKVPI-IZPVPAKOSA-N Thr-Thr-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O GRIUMVXCJDKVPI-IZPVPAKOSA-N 0.000 description 6
- BURPTJBFWIOHEY-UWJYBYFXSA-N Tyr-Ala-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 BURPTJBFWIOHEY-UWJYBYFXSA-N 0.000 description 6
- KHPLUFDSWGDRHD-SLFFLAALSA-N Tyr-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=C(C=C3)O)N)C(=O)O KHPLUFDSWGDRHD-SLFFLAALSA-N 0.000 description 6
- 108010008685 alanyl-glutamyl-aspartic acid Proteins 0.000 description 6
- 108010047857 aspartylglycine Proteins 0.000 description 6
- 108010092854 aspartyllysine Proteins 0.000 description 6
- 230000008878 coupling Effects 0.000 description 6
- 238000010168 coupling process Methods 0.000 description 6
- 238000005859 coupling reaction Methods 0.000 description 6
- 229940079593 drug Drugs 0.000 description 6
- 108010089804 glycyl-threonine Proteins 0.000 description 6
- 108010037850 glycylvaline Proteins 0.000 description 6
- 238000002823 phage display Methods 0.000 description 6
- 210000002966 serum Anatomy 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 125000004213 tert-butoxy group Chemical group [H]C([H])([H])C(O*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 6
- 108010061238 threonyl-glycine Proteins 0.000 description 6
- HNICLNKVURBTKV-NDEPHWFRSA-N (2s)-5-[[amino-[(2,2,4,6,7-pentamethyl-3h-1-benzofuran-5-yl)sulfonylamino]methylidene]amino]-2-(9h-fluoren-9-ylmethoxycarbonylamino)pentanoic acid Chemical compound C12=CC=CC=C2C2=CC=CC=C2C1COC(=O)N[C@H](C(O)=O)CCCN=C(N)NS(=O)(=O)C1=C(C)C(C)=C2OC(C)(C)CC2=C1C HNICLNKVURBTKV-NDEPHWFRSA-N 0.000 description 5
- DVWVZSJAYIJZFI-FXQIFTODSA-N Ala-Arg-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(O)=O DVWVZSJAYIJZFI-FXQIFTODSA-N 0.000 description 5
- CWEAKSWWKHGTRJ-BQBZGAKWSA-N Ala-Gly-Met Chemical compound [H]N[C@@H](C)C(=O)NCC(=O)N[C@@H](CCSC)C(O)=O CWEAKSWWKHGTRJ-BQBZGAKWSA-N 0.000 description 5
- OARAZORWIMYUPO-FXQIFTODSA-N Ala-Met-Cys Chemical compound CSCC[C@H](NC(=O)[C@H](C)N)C(=O)N[C@@H](CS)C(O)=O OARAZORWIMYUPO-FXQIFTODSA-N 0.000 description 5
- 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 5
- PGNNQOJOEGFAOR-KWQFWETISA-N Ala-Tyr-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=C(O)C=C1 PGNNQOJOEGFAOR-KWQFWETISA-N 0.000 description 5
- VKKYFICVTYKFIO-CIUDSAMLSA-N Arg-Ala-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCN=C(N)N VKKYFICVTYKFIO-CIUDSAMLSA-N 0.000 description 5
- ZTKHZAXGTFXUDD-VEVYYDQMSA-N Arg-Asn-Thr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ZTKHZAXGTFXUDD-VEVYYDQMSA-N 0.000 description 5
- PBSOQGZLPFVXPU-YUMQZZPRSA-N Arg-Glu-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O PBSOQGZLPFVXPU-YUMQZZPRSA-N 0.000 description 5
- ZEBDYGZVMMKZNB-SRVKXCTJSA-N Arg-Met-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CCCN=C(N)N)N ZEBDYGZVMMKZNB-SRVKXCTJSA-N 0.000 description 5
- 238000002965 ELISA Methods 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- JRCUFCXYZLPSDZ-ACZMJKKPSA-N Glu-Asp-Ser Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O JRCUFCXYZLPSDZ-ACZMJKKPSA-N 0.000 description 5
- PXXGVUVQWQGGIG-YUMQZZPRSA-N Glu-Gly-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCN=C(N)N PXXGVUVQWQGGIG-YUMQZZPRSA-N 0.000 description 5
- IWAXHBCACVWNHT-BQBZGAKWSA-N Gly-Asp-Arg Chemical compound NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N IWAXHBCACVWNHT-BQBZGAKWSA-N 0.000 description 5
- TZOVVRJYUDETQG-RCOVLWMOSA-N Gly-Asp-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CN TZOVVRJYUDETQG-RCOVLWMOSA-N 0.000 description 5
- FHQRLHFYVZAQHU-IUCAKERBSA-N Gly-Lys-Gln Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O FHQRLHFYVZAQHU-IUCAKERBSA-N 0.000 description 5
- IEGFSKKANYKBDU-QWHCGFSZSA-N Gly-Phe-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)NC(=O)CN)C(=O)O IEGFSKKANYKBDU-QWHCGFSZSA-N 0.000 description 5
- KSOBNUBCYHGUKH-UWVGGRQHSA-N Gly-Val-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)CN KSOBNUBCYHGUKH-UWVGGRQHSA-N 0.000 description 5
- WIPAMEKBSHNFQE-IUCAKERBSA-N Pro-Met-Gly Chemical compound CSCC[C@@H](C(=O)NCC(=O)O)NC(=O)[C@@H]1CCCN1 WIPAMEKBSHNFQE-IUCAKERBSA-N 0.000 description 5
- DMNANGOFEUVBRV-GJZGRUSLSA-N Pro-Trp-Gly Chemical compound N([C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)NCC(=O)O)C(=O)[C@@H]1CCCN1 DMNANGOFEUVBRV-GJZGRUSLSA-N 0.000 description 5
- 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 5
- RQXDSYQXBCRXBT-GUBZILKMSA-N Ser-Met-Arg Chemical compound OC[C@H](N)C(=O)N[C@@H](CCSC)C(=O)N[C@H](C(O)=O)CCCN=C(N)N RQXDSYQXBCRXBT-GUBZILKMSA-N 0.000 description 5
- HHJFMHQYEAAOBM-ZLUOBGJFSA-N Ser-Ser-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O HHJFMHQYEAAOBM-ZLUOBGJFSA-N 0.000 description 5
- OLKICIBQRVSQMA-SRVKXCTJSA-N Ser-Ser-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O OLKICIBQRVSQMA-SRVKXCTJSA-N 0.000 description 5
- WFUAUEQXPVNAEF-ZJDVBMNYSA-N Thr-Arg-Thr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)O)C(O)=O)CCCN=C(N)N WFUAUEQXPVNAEF-ZJDVBMNYSA-N 0.000 description 5
- YJCVECXVYHZOBK-KNZXXDILSA-N Thr-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H]([C@@H](C)O)N YJCVECXVYHZOBK-KNZXXDILSA-N 0.000 description 5
- YOPQYBJJNSIQGZ-JNPHEJMOSA-N Thr-Tyr-Tyr Chemical compound C([C@H](NC(=O)[C@@H](N)[C@H](O)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 YOPQYBJJNSIQGZ-JNPHEJMOSA-N 0.000 description 5
- HKYTWJOWZTWBQB-AVGNSLFASA-N Tyr-Glu-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 HKYTWJOWZTWBQB-AVGNSLFASA-N 0.000 description 5
- UDLYXGYWTVOIKU-QXEWZRGKSA-N Val-Asn-Arg Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N UDLYXGYWTVOIKU-QXEWZRGKSA-N 0.000 description 5
- 108010068265 aspartyltyrosine Proteins 0.000 description 5
- 238000010494 dissociation reaction Methods 0.000 description 5
- 230000005593 dissociations Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 230000002163 immunogen Effects 0.000 description 5
- 238000002347 injection Methods 0.000 description 5
- 239000007924 injection Substances 0.000 description 5
- 108010031719 prolyl-serine Proteins 0.000 description 5
- 108010053725 prolylvaline Proteins 0.000 description 5
- 238000006467 substitution reaction Methods 0.000 description 5
- 210000004881 tumor cell Anatomy 0.000 description 5
- 108010047303 von Willebrand Factor Proteins 0.000 description 5
- 102100036537 von Willebrand factor Human genes 0.000 description 5
- HLCTYBOTPCIHTG-QMMMGPOBSA-N (2r)-3-(acetamidomethylsulfanyl)-2-[(2-methylpropan-2-yl)oxycarbonylamino]propanoic acid Chemical compound CC(=O)NCSC[C@@H](C(O)=O)NC(=O)OC(C)(C)C HLCTYBOTPCIHTG-QMMMGPOBSA-N 0.000 description 4
- GVIXTVCDNCXXSH-AWEZNQCLSA-N (2s)-2-amino-5-[[amino-[(2,2,4,6,7-pentamethyl-3h-1-benzofuran-5-yl)sulfonylamino]methylidene]amino]pentanoic acid Chemical compound OC(=O)[C@@H](N)CCCN=C(N)NS(=O)(=O)C1=C(C)C(C)=C2OC(C)(C)CC2=C1C GVIXTVCDNCXXSH-AWEZNQCLSA-N 0.000 description 4
- 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 4
- FRBAHXABMQXSJQ-FXQIFTODSA-N Arg-Ser-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O FRBAHXABMQXSJQ-FXQIFTODSA-N 0.000 description 4
- QHUOOCKNNURZSL-IHRRRGAJSA-N Arg-Tyr-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(O)=O QHUOOCKNNURZSL-IHRRRGAJSA-N 0.000 description 4
- 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 4
- NAPNAGZWHQHZLG-ZLUOBGJFSA-N Asp-Asp-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)O)N NAPNAGZWHQHZLG-ZLUOBGJFSA-N 0.000 description 4
- 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 4
- CGYDXNKRIMJMLV-GUBZILKMSA-N Glu-Arg-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O CGYDXNKRIMJMLV-GUBZILKMSA-N 0.000 description 4
- OGCIHJPYKVSMTE-YUMQZZPRSA-N Gly-Arg-Glu Chemical compound [H]NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O OGCIHJPYKVSMTE-YUMQZZPRSA-N 0.000 description 4
- OMOZPGCHVWOXHN-BQBZGAKWSA-N Gly-Met-Ser Chemical compound CSCC[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)CN OMOZPGCHVWOXHN-BQBZGAKWSA-N 0.000 description 4
- NXRNRBOKDBIVKQ-CXTHYWKRSA-N Ile-Tyr-Tyr Chemical compound CC[C@H](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 NXRNRBOKDBIVKQ-CXTHYWKRSA-N 0.000 description 4
- 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 4
- QUCDKEKDPYISNX-HJGDQZAQSA-N Lys-Asn-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QUCDKEKDPYISNX-HJGDQZAQSA-N 0.000 description 4
- CHLJXFMOQGYDNH-SZMVWBNQSA-N Met-Arg-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](N)CCSC)C(O)=O)=CNC2=C1 CHLJXFMOQGYDNH-SZMVWBNQSA-N 0.000 description 4
- MHQXIBRPDKXDGZ-ZFWWWQNUSA-N Met-Gly-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)CNC(=O)[C@@H](N)CCSC)C(O)=O)=CNC2=C1 MHQXIBRPDKXDGZ-ZFWWWQNUSA-N 0.000 description 4
- 241000699670 Mus sp. Species 0.000 description 4
- WUGMRIBZSVSJNP-UHFFFAOYSA-N N-L-alanyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C)C(O)=O)=CNC2=C1 WUGMRIBZSVSJNP-UHFFFAOYSA-N 0.000 description 4
- 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 4
- 206010028980 Neoplasm Diseases 0.000 description 4
- 101150003085 Pdcl gene Proteins 0.000 description 4
- ZAUHSLVPDLNTRZ-QXEWZRGKSA-N Pro-Val-Asn Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O ZAUHSLVPDLNTRZ-QXEWZRGKSA-N 0.000 description 4
- JFWDJFULOLKQFY-QWRGUYRKSA-N Ser-Gly-Phe Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O JFWDJFULOLKQFY-QWRGUYRKSA-N 0.000 description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 4
- 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 4
- ABWNZPOIUJMNKT-IXOXFDKPSA-N Thr-Phe-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O ABWNZPOIUJMNKT-IXOXFDKPSA-N 0.000 description 4
- UGFOSENEZHEQKX-PJODQICGSA-N Trp-Val-Ala Chemical compound CC(C)[C@H](NC(=O)[C@@H](N)Cc1c[nH]c2ccccc12)C(=O)N[C@@H](C)C(O)=O UGFOSENEZHEQKX-PJODQICGSA-N 0.000 description 4
- VTFWAGGJDRSQFG-MELADBBJSA-N Tyr-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC2=CC=C(C=C2)O)N)C(=O)O VTFWAGGJDRSQFG-MELADBBJSA-N 0.000 description 4
- SMLCYZYQFRTLCO-UWJYBYFXSA-N Tyr-Cys-Ala Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CS)C(=O)N[C@@H](C)C(O)=O SMLCYZYQFRTLCO-UWJYBYFXSA-N 0.000 description 4
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 4
- KPFBUSLHFFWMAI-HYRPPVSQSA-N [(8r,9s,10r,13s,14s,17r)-17-acetyl-6-formyl-3-methoxy-10,13-dimethyl-1,2,7,8,9,11,12,14,15,16-decahydrocyclopenta[a]phenanthren-17-yl] acetate Chemical compound C1C[C@@H]2[C@](CCC(OC)=C3)(C)C3=C(C=O)C[C@H]2[C@@H]2CC[C@](OC(C)=O)(C(C)=O)[C@]21C KPFBUSLHFFWMAI-HYRPPVSQSA-N 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- LBSPZZSGTIBOFG-UHFFFAOYSA-N bis[2-(4,5-dihydro-1h-imidazol-2-yl)propan-2-yl]diazene;dihydrochloride Chemical compound Cl.Cl.N=1CCNC=1C(C)(C)N=NC(C)(C)C1=NCCN1 LBSPZZSGTIBOFG-UHFFFAOYSA-N 0.000 description 4
- 238000009833 condensation Methods 0.000 description 4
- 230000005494 condensation Effects 0.000 description 4
- 238000010276 construction Methods 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 238000001727 in vivo Methods 0.000 description 4
- 230000003993 interaction Effects 0.000 description 4
- 108010009298 lysylglutamic acid Proteins 0.000 description 4
- 108010064235 lysylglycine Proteins 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 230000035772 mutation Effects 0.000 description 4
- 238000010647 peptide synthesis reaction Methods 0.000 description 4
- 239000008194 pharmaceutical composition Substances 0.000 description 4
- 125000006239 protecting group Chemical group 0.000 description 4
- 108010071207 serylmethionine Proteins 0.000 description 4
- 230000008685 targeting Effects 0.000 description 4
- 230000001225 therapeutic effect Effects 0.000 description 4
- 108010044292 tryptophyltyrosine Proteins 0.000 description 4
- VVQIIIAZJXTLRE-QMMMGPOBSA-N (2s)-2-amino-6-[(2-methylpropan-2-yl)oxycarbonylamino]hexanoic acid Chemical compound CC(C)(C)OC(=O)NCCCC[C@H](N)C(O)=O VVQIIIAZJXTLRE-QMMMGPOBSA-N 0.000 description 3
- RLMISHABBKUNFO-WHFBIAKZSA-N Ala-Ala-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O RLMISHABBKUNFO-WHFBIAKZSA-N 0.000 description 3
- NHCPCLJZRSIDHS-ZLUOBGJFSA-N Ala-Asp-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O NHCPCLJZRSIDHS-ZLUOBGJFSA-N 0.000 description 3
- LMFXXZPPZDCPTA-ZKWXMUAHSA-N Ala-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@H](C)N LMFXXZPPZDCPTA-ZKWXMUAHSA-N 0.000 description 3
- OMDNCNKNEGFOMM-BQBZGAKWSA-N Ala-Met-Gly Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)NCC(O)=O OMDNCNKNEGFOMM-BQBZGAKWSA-N 0.000 description 3
- PRLPSDIHSRITSF-UNQGMJICSA-N Arg-Phe-Thr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PRLPSDIHSRITSF-UNQGMJICSA-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
- 241000588724 Escherichia coli Species 0.000 description 3
- QLNKFGTZOBVMCS-JBACZVJFSA-N Glu-Tyr-Trp Chemical compound [H]N[C@@H](CCC(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 QLNKFGTZOBVMCS-JBACZVJFSA-N 0.000 description 3
- NZAFOTBEULLEQB-WDSKDSINSA-N Gly-Asn-Glu Chemical compound C(CC(=O)O)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)CN NZAFOTBEULLEQB-WDSKDSINSA-N 0.000 description 3
- VNNRLUNBJSWZPF-ZKWXMUAHSA-N Gly-Ser-Ile Chemical compound [H]NCC(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VNNRLUNBJSWZPF-ZKWXMUAHSA-N 0.000 description 3
- 108010093488 His-His-His-His-His-His Proteins 0.000 description 3
- CKSXSQUVEYCDIW-AVGNSLFASA-N Lys-Arg-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCCN)N CKSXSQUVEYCDIW-AVGNSLFASA-N 0.000 description 3
- PESQCPHRXOFIPX-UHFFFAOYSA-N N-L-methionyl-L-tyrosine Natural products CSCCC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 PESQCPHRXOFIPX-UHFFFAOYSA-N 0.000 description 3
- 102000035195 Peptidases Human genes 0.000 description 3
- 108091005804 Peptidases Proteins 0.000 description 3
- DFEVBOYEUQJGER-JURCDPSOSA-N Phe-Ala-Ile Chemical compound N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O DFEVBOYEUQJGER-JURCDPSOSA-N 0.000 description 3
- JXQVYPWVGUOIDV-MXAVVETBSA-N Phe-Ser-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JXQVYPWVGUOIDV-MXAVVETBSA-N 0.000 description 3
- GFHXZNVJIKMAGO-IHRRRGAJSA-N Pro-Phe-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O GFHXZNVJIKMAGO-IHRRRGAJSA-N 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 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 3
- WBINSDOPZHQPPM-AVGNSLFASA-N Ser-Glu-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CO)N)O WBINSDOPZHQPPM-AVGNSLFASA-N 0.000 description 3
- 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 3
- BDMWLJLPPUCLNV-XGEHTFHBSA-N Ser-Thr-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O BDMWLJLPPUCLNV-XGEHTFHBSA-N 0.000 description 3
- UBTNVMGPMYDYIU-HJPIBITLSA-N Ser-Tyr-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O UBTNVMGPMYDYIU-HJPIBITLSA-N 0.000 description 3
- BEBVVQPDSHHWQL-NRPADANISA-N Ser-Val-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O BEBVVQPDSHHWQL-NRPADANISA-N 0.000 description 3
- YLXAMFZYJTZXFH-OLHMAJIHSA-N Thr-Asn-Asp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N)O YLXAMFZYJTZXFH-OLHMAJIHSA-N 0.000 description 3
- XPNSAQMEAVSQRD-FBCQKBJTSA-N Thr-Gly-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)NCC(O)=O XPNSAQMEAVSQRD-FBCQKBJTSA-N 0.000 description 3
- NCXVJIQMWSGRHY-KXNHARMFSA-N Thr-Leu-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N)O NCXVJIQMWSGRHY-KXNHARMFSA-N 0.000 description 3
- XIHGJKFSIDTDKV-LYARXQMPSA-N Thr-Phe-Trp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O XIHGJKFSIDTDKV-LYARXQMPSA-N 0.000 description 3
- XJPXTYLVMUZGNW-IHRRRGAJSA-N Tyr-Pro-Asp Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O XJPXTYLVMUZGNW-IHRRRGAJSA-N 0.000 description 3
- MWUYSCVVPVITMW-IGNZVWTISA-N Tyr-Tyr-Ala Chemical compound C([C@@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 MWUYSCVVPVITMW-IGNZVWTISA-N 0.000 description 3
- MDYSKHBSPXUOPV-JSGCOSHPSA-N Val-Gly-Phe Chemical compound CC(C)[C@@H](C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N MDYSKHBSPXUOPV-JSGCOSHPSA-N 0.000 description 3
- 102000009524 Vascular Endothelial Growth Factor A Human genes 0.000 description 3
- 238000007792 addition Methods 0.000 description 3
- 239000000654 additive Substances 0.000 description 3
- 230000000996 additive effect Effects 0.000 description 3
- 239000002246 antineoplastic agent Substances 0.000 description 3
- 230000004071 biological effect Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 238000003018 immunoassay Methods 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 239000003112 inhibitor Substances 0.000 description 3
- 229910052751 metal Inorganic materials 0.000 description 3
- 239000002184 metal Substances 0.000 description 3
- 108010051242 phenylalanylserine Proteins 0.000 description 3
- 229920001223 polyethylene glycol Polymers 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000007790 solid phase Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 description 3
- VYMPLPIFKRHAAC-UHFFFAOYSA-N 1,2-ethanedithiol Chemical compound SCCS VYMPLPIFKRHAAC-UHFFFAOYSA-N 0.000 description 2
- 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 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- YIRHQVZRKURGRG-UHFFFAOYSA-N 3-(9H-fluoren-9-ylmethoxycarbonylamino)-4-(methylamino)benzoic acid Chemical compound CNc1ccc(cc1NC(=O)OCC1c2ccccc2-c2ccccc12)C(O)=O YIRHQVZRKURGRG-UHFFFAOYSA-N 0.000 description 2
- LRLZBKSOHQLVRG-UHFFFAOYSA-N 4-amino-3-(9h-fluoren-9-ylmethoxycarbonylamino)benzoic acid Chemical compound NC1=CC=C(C(O)=O)C=C1NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 LRLZBKSOHQLVRG-UHFFFAOYSA-N 0.000 description 2
- DKJPOZOEBONHFS-ZLUOBGJFSA-N Ala-Ala-Asp Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O DKJPOZOEBONHFS-ZLUOBGJFSA-N 0.000 description 2
- MPLOSMWGDNJSEV-WHFBIAKZSA-N Ala-Gly-Asp Chemical compound [H]N[C@@H](C)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O MPLOSMWGDNJSEV-WHFBIAKZSA-N 0.000 description 2
- IFKQPMZRDQZSHI-GHCJXIJMSA-N Ala-Ile-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O IFKQPMZRDQZSHI-GHCJXIJMSA-N 0.000 description 2
- VNFSAYFQLXPHPY-CIQUZCHMSA-N Ala-Thr-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VNFSAYFQLXPHPY-CIQUZCHMSA-N 0.000 description 2
- KTXKIYXZQFWJKB-VZFHVOOUSA-N Ala-Thr-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O KTXKIYXZQFWJKB-VZFHVOOUSA-N 0.000 description 2
- VVJTWSRNMJNDPN-IUCAKERBSA-N Arg-Met-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)NCC(O)=O VVJTWSRNMJNDPN-IUCAKERBSA-N 0.000 description 2
- 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 2
- JPPLRQVZMZFOSX-UWJYBYFXSA-N Asn-Tyr-Ala Chemical compound NC(=O)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=C(O)C=C1 JPPLRQVZMZFOSX-UWJYBYFXSA-N 0.000 description 2
- YSYTWUMRHSFODC-QWRGUYRKSA-N Asn-Tyr-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O YSYTWUMRHSFODC-QWRGUYRKSA-N 0.000 description 2
- RDRMWJBLOSRRAW-BYULHYEWSA-N Asp-Asn-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O RDRMWJBLOSRRAW-BYULHYEWSA-N 0.000 description 2
- SAKCBXNPWDRWPE-BQBZGAKWSA-N Asp-Met-Gly Chemical compound CSCC[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CC(=O)O)N SAKCBXNPWDRWPE-BQBZGAKWSA-N 0.000 description 2
- MFDPBZAFCRKYEY-LAEOZQHASA-N Asp-Val-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O MFDPBZAFCRKYEY-LAEOZQHASA-N 0.000 description 2
- 241000282832 Camelidae Species 0.000 description 2
- 241000282836 Camelus dromedarius Species 0.000 description 2
- GOKFTBDYUJCCSN-QEJZJMRPSA-N Cys-Glu-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CS)N GOKFTBDYUJCCSN-QEJZJMRPSA-N 0.000 description 2
- GUKYYUFHWYRMEU-WHFBIAKZSA-N Cys-Gly-Asp Chemical compound [H]N[C@@H](CS)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O GUKYYUFHWYRMEU-WHFBIAKZSA-N 0.000 description 2
- KVGPYKUIHZJWGA-BQBZGAKWSA-N Cys-Met-Gly Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCSC)C(=O)NCC(O)=O KVGPYKUIHZJWGA-BQBZGAKWSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- NSORZJXKUQFEKL-JGVFFNPUSA-N Gln-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CCC(=O)N)N)C(=O)O NSORZJXKUQFEKL-JGVFFNPUSA-N 0.000 description 2
- XBWGJWXGUNSZAT-CIUDSAMLSA-N Gln-Met-Asp Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CCC(=O)N)N XBWGJWXGUNSZAT-CIUDSAMLSA-N 0.000 description 2
- YWAQATDNEKZFFK-BYPYZUCNSA-N Gly-Gly-Ser Chemical compound NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O YWAQATDNEKZFFK-BYPYZUCNSA-N 0.000 description 2
- 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 2
- LHYJCVCQPWRMKZ-WEDXCCLWSA-N Gly-Leu-Thr Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LHYJCVCQPWRMKZ-WEDXCCLWSA-N 0.000 description 2
- BXDLTKLPPKBVEL-FJXKBIBVSA-N Gly-Thr-Met Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(O)=O BXDLTKLPPKBVEL-FJXKBIBVSA-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
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- CDGLBYSAZFIIJO-RCOVLWMOSA-N Ile-Gly-Gly Chemical compound CC[C@H](C)[C@H]([NH3+])C(=O)NCC(=O)NCC([O-])=O CDGLBYSAZFIIJO-RCOVLWMOSA-N 0.000 description 2
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 2
- CQQGCWPXDHTTNF-GUBZILKMSA-N Leu-Ala-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCC(O)=O CQQGCWPXDHTTNF-GUBZILKMSA-N 0.000 description 2
- PRZVBIAOPFGAQF-SRVKXCTJSA-N Leu-Glu-Met Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O PRZVBIAOPFGAQF-SRVKXCTJSA-N 0.000 description 2
- GZRABTMNWJXFMH-UVOCVTCTSA-N Leu-Thr-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GZRABTMNWJXFMH-UVOCVTCTSA-N 0.000 description 2
- YIRIDPUGZKHMHT-ACRUOGEOSA-N Leu-Tyr-Tyr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O YIRIDPUGZKHMHT-ACRUOGEOSA-N 0.000 description 2
- PRCHKVGXZVTALR-KKUMJFAQSA-N Lys-His-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)NC(=O)[C@H](CCCCN)N PRCHKVGXZVTALR-KKUMJFAQSA-N 0.000 description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 2
- CAODKDAPYGUMLK-FXQIFTODSA-N Met-Asn-Ser Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O CAODKDAPYGUMLK-FXQIFTODSA-N 0.000 description 2
- GHQFLTYXGUETFD-UFYCRDLUSA-N Met-Tyr-Tyr Chemical compound CSCC[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 GHQFLTYXGUETFD-UFYCRDLUSA-N 0.000 description 2
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 2
- JNRFYJZCMHHGMH-UBHSHLNASA-N Phe-Ala-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=CC=C1 JNRFYJZCMHHGMH-UBHSHLNASA-N 0.000 description 2
- GXDPQJUBLBZKDY-IAVJCBSLSA-N Phe-Ile-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O GXDPQJUBLBZKDY-IAVJCBSLSA-N 0.000 description 2
- 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 2
- 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 2
- 229920001213 Polysorbate 20 Polymers 0.000 description 2
- AFXCXDQNRXTSBD-FJXKBIBVSA-N Pro-Gly-Thr Chemical compound [H]N1CCC[C@H]1C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O AFXCXDQNRXTSBD-FJXKBIBVSA-N 0.000 description 2
- VEUACYMXJKXALX-IHRRRGAJSA-N Pro-Tyr-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(O)=O VEUACYMXJKXALX-IHRRRGAJSA-N 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- INCNPLPRPOYTJI-JBDRJPRFSA-N Ser-Cys-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CO)N INCNPLPRPOYTJI-JBDRJPRFSA-N 0.000 description 2
- 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 2
- VXYQOFXBIXKPCX-BQBZGAKWSA-N Ser-Met-Gly Chemical compound CSCC[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CO)N VXYQOFXBIXKPCX-BQBZGAKWSA-N 0.000 description 2
- XZKQVQKUZMAADP-IMJSIDKUSA-N Ser-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(O)=O XZKQVQKUZMAADP-IMJSIDKUSA-N 0.000 description 2
- 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 2
- BCAVNDNYOGTQMQ-AAEUAGOBSA-N Ser-Trp-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)NCC(O)=O BCAVNDNYOGTQMQ-AAEUAGOBSA-N 0.000 description 2
- ZWSZBWAFDZRBNM-UBHSHLNASA-N Ser-Trp-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CO)C(O)=O ZWSZBWAFDZRBNM-UBHSHLNASA-N 0.000 description 2
- YXGCIEUDOHILKR-IHRRRGAJSA-N Ser-Tyr-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CO)N YXGCIEUDOHILKR-IHRRRGAJSA-N 0.000 description 2
- 210000001744 T-lymphocyte Anatomy 0.000 description 2
- 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 2
- OHAJHDJOCKKJLV-LKXGYXEUSA-N Thr-Asp-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O OHAJHDJOCKKJLV-LKXGYXEUSA-N 0.000 description 2
- ZBKDBZUTTXINIX-RWRJDSDZSA-N Thr-Ile-Gln Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(N)=O)C(O)=O ZBKDBZUTTXINIX-RWRJDSDZSA-N 0.000 description 2
- SJWLQICJOBMOGG-PMVMPFDFSA-N Trp-Tyr-His Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC3=CC=C(C=C3)O)C(=O)N[C@@H](CC4=CN=CN4)C(=O)O)N SJWLQICJOBMOGG-PMVMPFDFSA-N 0.000 description 2
- NMKJPMCEKQHRPD-IRXDYDNUSA-N Tyr-Gly-Tyr Chemical compound C([C@H](N)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 NMKJPMCEKQHRPD-IRXDYDNUSA-N 0.000 description 2
- 108010064997 VPY tripeptide Proteins 0.000 description 2
- WOCYUGQDXPTQPY-FXQIFTODSA-N Val-Ala-Cys Chemical compound C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](C(C)C)N WOCYUGQDXPTQPY-FXQIFTODSA-N 0.000 description 2
- DIOSYUIWOQCXNR-ONGXEEELSA-N Val-Lys-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O DIOSYUIWOQCXNR-ONGXEEELSA-N 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 230000002411 adverse Effects 0.000 description 2
- 230000009824 affinity maturation Effects 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- 235000004279 alanine Nutrition 0.000 description 2
- 108010041407 alanylaspartic acid Proteins 0.000 description 2
- 150000001412 amines Chemical group 0.000 description 2
- 230000030741 antigen processing and presentation Effects 0.000 description 2
- 210000000612 antigen-presenting cell Anatomy 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000010382 chemical cross-linking Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000002983 circular dichroism Methods 0.000 description 2
- 238000000978 circular dichroism spectroscopy Methods 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 239000000562 conjugate Substances 0.000 description 2
- 230000021615 conjugation Effects 0.000 description 2
- 239000003431 cross linking reagent Substances 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 238000006477 desulfuration reaction Methods 0.000 description 2
- 230000023556 desulfurization Effects 0.000 description 2
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 2
- 239000000539 dimer Substances 0.000 description 2
- 238000009509 drug development Methods 0.000 description 2
- 238000011067 equilibration Methods 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 229910052731 fluorine Inorganic materials 0.000 description 2
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 2
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 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
- 108010087823 glycyltyrosine Proteins 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 230000003053 immunization Effects 0.000 description 2
- 238000002649 immunization Methods 0.000 description 2
- 238000003364 immunohistochemistry Methods 0.000 description 2
- 238000001114 immunoprecipitation Methods 0.000 description 2
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- 239000000825 pharmaceutical preparation Substances 0.000 description 2
- 229940127557 pharmaceutical product Drugs 0.000 description 2
- 229910052698 phosphorus Inorganic materials 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 2
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 2
- 229910052700 potassium Inorganic materials 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 108010029020 prolylglycine Proteins 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 229940076372 protein antagonist Drugs 0.000 description 2
- 238000003127 radioimmunoassay Methods 0.000 description 2
- 108010026333 seryl-proline Proteins 0.000 description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 230000006641 stabilisation Effects 0.000 description 2
- 238000011105 stabilization Methods 0.000 description 2
- 229910052717 sulfur Inorganic materials 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 229910052721 tungsten Inorganic materials 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N urea group Chemical group NC(=O)N XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 229910052727 yttrium Inorganic materials 0.000 description 2
- XUDGDVPXDYGCTG-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) 2-[2-(2,5-dioxopyrrolidin-1-yl)oxycarbonyloxyethylsulfonyl]ethyl carbonate Chemical compound O=C1CCC(=O)N1OC(=O)OCCS(=O)(=O)CCOC(=O)ON1C(=O)CCC1=O XUDGDVPXDYGCTG-UHFFFAOYSA-N 0.000 description 1
- JAUKCFULLJFBFN-RUZDIDTESA-N (2r)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-[4-[(2-methylpropan-2-yl)oxy]phenyl]propanoic acid Chemical compound C1=CC(OC(C)(C)C)=CC=C1C[C@H](C(O)=O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 JAUKCFULLJFBFN-RUZDIDTESA-N 0.000 description 1
- HNICLNKVURBTKV-MUUNZHRXSA-N (2r)-5-[[amino-[(2,2,4,6,7-pentamethyl-3h-1-benzofuran-5-yl)sulfonylamino]methylidene]amino]-2-(9h-fluoren-9-ylmethoxycarbonylamino)pentanoic acid Chemical compound C12=CC=CC=C2C2=CC=CC=C2C1COC(=O)N[C@@H](C(O)=O)CCCN=C(N)NS(=O)(=O)C1=C(C)C(C)=C2OC(C)(C)CC2=C1C HNICLNKVURBTKV-MUUNZHRXSA-N 0.000 description 1
- REITVGIIZHFVGU-IBGZPJMESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-[(2-methylpropan-2-yl)oxy]propanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](COC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 REITVGIIZHFVGU-IBGZPJMESA-N 0.000 description 1
- JAUKCFULLJFBFN-VWLOTQADSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-[4-[(2-methylpropan-2-yl)oxy]phenyl]propanoic acid Chemical compound C1=CC(OC(C)(C)C)=CC=C1C[C@@H](C(O)=O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 JAUKCFULLJFBFN-VWLOTQADSA-N 0.000 description 1
- IGXNPQWXIRIGBF-KEOOTSPTSA-N (2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-amino-3-(1h-imidazol-5-yl)propanoyl]amino]-3-(1h-imidazol-5-yl)propanoyl]amino]-3-(1h-imidazol-5-yl)propanoyl]amino]-3-(1h-imidazol-5-yl)propanoyl]amino]-3-(1h-imidazol-5-yl)propanoic acid Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC=1NC=NC=1)C(O)=O)C1=CN=CN1 IGXNPQWXIRIGBF-KEOOTSPTSA-N 0.000 description 1
- DYWUPCCKOVTCFZ-LBPRGKRZSA-N (2s)-2-amino-3-[1-[(2-methylpropan-2-yl)oxycarbonyl]indol-3-yl]propanoic acid Chemical compound C1=CC=C2N(C(=O)OC(C)(C)C)C=C(C[C@H](N)C(O)=O)C2=C1 DYWUPCCKOVTCFZ-LBPRGKRZSA-N 0.000 description 1
- QGKMIGUHVLGJBR-UHFFFAOYSA-M (4z)-1-(3-methylbutyl)-4-[[1-(3-methylbutyl)quinolin-1-ium-4-yl]methylidene]quinoline;iodide Chemical compound [I-].C12=CC=CC=C2N(CCC(C)C)C=CC1=CC1=CC=[N+](CCC(C)C)C2=CC=CC=C12 QGKMIGUHVLGJBR-UHFFFAOYSA-M 0.000 description 1
- VOTJUWBJENROFB-UHFFFAOYSA-N 1-[3-[[3-(2,5-dioxo-3-sulfopyrrolidin-1-yl)oxy-3-oxopropyl]disulfanyl]propanoyloxy]-2,5-dioxopyrrolidine-3-sulfonic acid Chemical compound O=C1C(S(=O)(=O)O)CC(=O)N1OC(=O)CCSSCCC(=O)ON1C(=O)C(S(O)(=O)=O)CC1=O VOTJUWBJENROFB-UHFFFAOYSA-N 0.000 description 1
- WQQBUTMELIQJNY-UHFFFAOYSA-N 1-[4-(2,5-dioxo-3-sulfopyrrolidin-1-yl)oxy-2,3-dihydroxy-4-oxobutanoyl]oxy-2,5-dioxopyrrolidine-3-sulfonic acid Chemical compound O=C1CC(S(O)(=O)=O)C(=O)N1OC(=O)C(O)C(O)C(=O)ON1C(=O)CC(S(O)(=O)=O)C1=O WQQBUTMELIQJNY-UHFFFAOYSA-N 0.000 description 1
- UKPGFKQVRITNFM-KBPBESRZSA-N 2-[[2-[[(2s)-1-[(2s)-2-amino-3-(4-hydroxyphenyl)propanoyl]pyrrolidine-2-carbonyl]amino]acetyl]amino]acetic acid Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)NCC(=O)NCC(O)=O)C1=CC=C(O)C=C1 UKPGFKQVRITNFM-KBPBESRZSA-N 0.000 description 1
- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 description 1
- TVZRAEYQIKYCPH-UHFFFAOYSA-N 3-(trimethylsilyl)propane-1-sulfonic acid Chemical compound C[Si](C)(C)CCCS(O)(=O)=O TVZRAEYQIKYCPH-UHFFFAOYSA-N 0.000 description 1
- QLHLYJHNOCILIT-UHFFFAOYSA-N 4-o-(2,5-dioxopyrrolidin-1-yl) 1-o-[2-[4-(2,5-dioxopyrrolidin-1-yl)oxy-4-oxobutanoyl]oxyethyl] butanedioate Chemical compound O=C1CCC(=O)N1OC(=O)CCC(=O)OCCOC(=O)CCC(=O)ON1C(=O)CCC1=O QLHLYJHNOCILIT-UHFFFAOYSA-N 0.000 description 1
- HBAQYPYDRFILMT-UHFFFAOYSA-N 8-[3-(1-cyclopropylpyrazol-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl]-3-methyl-3,8-diazabicyclo[3.2.1]octan-2-one Chemical class C1(CC1)N1N=CC(=C1)C1=NNC2=C1N=C(N=C2)N1C2C(N(CC1CC2)C)=O HBAQYPYDRFILMT-UHFFFAOYSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- SBGXWWCLHIOABR-UHFFFAOYSA-N Ala Ala Gly Ala Chemical compound CC(N)C(=O)NC(C)C(=O)NCC(=O)NC(C)C(O)=O SBGXWWCLHIOABR-UHFFFAOYSA-N 0.000 description 1
- HHGYNJRJIINWAK-FXQIFTODSA-N Ala-Ala-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N HHGYNJRJIINWAK-FXQIFTODSA-N 0.000 description 1
- LGQPPBQRUBVTIF-JBDRJPRFSA-N Ala-Ala-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O LGQPPBQRUBVTIF-JBDRJPRFSA-N 0.000 description 1
- VBDMWOKJZDCFJM-FXQIFTODSA-N Ala-Ala-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](C)N VBDMWOKJZDCFJM-FXQIFTODSA-N 0.000 description 1
- JBVSSSZFNTXJDX-YTLHQDLWSA-N Ala-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](C)N JBVSSSZFNTXJDX-YTLHQDLWSA-N 0.000 description 1
- KQFRUSHJPKXBMB-BHDSKKPTSA-N Ala-Ala-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](C)NC(=O)[C@@H](N)C)C(O)=O)=CNC2=C1 KQFRUSHJPKXBMB-BHDSKKPTSA-N 0.000 description 1
- DWINFPQUSSHSFS-UVBJJODRSA-N Ala-Arg-Trp Chemical compound N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CNC2=CC=CC=C12)C(=O)O DWINFPQUSSHSFS-UVBJJODRSA-N 0.000 description 1
- WYPUMLRSQMKIJU-BPNCWPANSA-N Ala-Arg-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O WYPUMLRSQMKIJU-BPNCWPANSA-N 0.000 description 1
- WXERCAHAIKMTKX-ZLUOBGJFSA-N Ala-Asp-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O WXERCAHAIKMTKX-ZLUOBGJFSA-N 0.000 description 1
- FVSOUJZKYWEFOB-KBIXCLLPSA-N Ala-Gln-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](C)N FVSOUJZKYWEFOB-KBIXCLLPSA-N 0.000 description 1
- ZDYNWWQXFRUOEO-XDTLVQLUSA-N Ala-Gln-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZDYNWWQXFRUOEO-XDTLVQLUSA-N 0.000 description 1
- 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 1
- OMMDTNGURYRDAC-NRPADANISA-N Ala-Glu-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O OMMDTNGURYRDAC-NRPADANISA-N 0.000 description 1
- DVJSJDDYCYSMFR-ZKWXMUAHSA-N Ala-Ile-Gly Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O DVJSJDDYCYSMFR-ZKWXMUAHSA-N 0.000 description 1
- LXAARTARZJJCMB-CIQUZCHMSA-N Ala-Ile-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LXAARTARZJJCMB-CIQUZCHMSA-N 0.000 description 1
- QQACQIHVWCVBBR-GVARAGBVSA-N Ala-Ile-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O QQACQIHVWCVBBR-GVARAGBVSA-N 0.000 description 1
- 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 1
- PMQXMXAASGFUDX-SRVKXCTJSA-N Ala-Lys-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](C)N)CCCCN PMQXMXAASGFUDX-SRVKXCTJSA-N 0.000 description 1
- CHFFHQUVXHEGBY-GARJFASQSA-N Ala-Lys-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@@H]1C(=O)O)N CHFFHQUVXHEGBY-GARJFASQSA-N 0.000 description 1
- NINQYGGNRIBFSC-CIUDSAMLSA-N Ala-Lys-Ser Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@@H](CO)C(O)=O NINQYGGNRIBFSC-CIUDSAMLSA-N 0.000 description 1
- XUCHENWTTBFODJ-FXQIFTODSA-N Ala-Met-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C)C(O)=O XUCHENWTTBFODJ-FXQIFTODSA-N 0.000 description 1
- VHEVVUZDDUCAKU-FXQIFTODSA-N Ala-Met-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(O)=O VHEVVUZDDUCAKU-FXQIFTODSA-N 0.000 description 1
- FVNAUOZKIPAYNA-BPNCWPANSA-N Ala-Met-Tyr Chemical compound CSCC[C@H](NC(=O)[C@H](C)N)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 FVNAUOZKIPAYNA-BPNCWPANSA-N 0.000 description 1
- RUXQNKVQSKOOBS-JURCDPSOSA-N Ala-Phe-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O RUXQNKVQSKOOBS-JURCDPSOSA-N 0.000 description 1
- BTRULDJUUVGRNE-DCAQKATOSA-N Ala-Pro-Lys Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(O)=O BTRULDJUUVGRNE-DCAQKATOSA-N 0.000 description 1
- NZGRHTKZFSVPAN-BIIVOSGPSA-N Ala-Ser-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)N1CCC[C@@H]1C(=O)O)N NZGRHTKZFSVPAN-BIIVOSGPSA-N 0.000 description 1
- MUGAESARFRGOTQ-IGNZVWTISA-N Ala-Tyr-Tyr Chemical compound 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 MUGAESARFRGOTQ-IGNZVWTISA-N 0.000 description 1
- VHAQSYHSDKERBS-XPUUQOCRSA-N Ala-Val-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O VHAQSYHSDKERBS-XPUUQOCRSA-N 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- DBKNLHKEVPZVQC-LPEHRKFASA-N Arg-Ala-Pro Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@@H]1C(O)=O DBKNLHKEVPZVQC-LPEHRKFASA-N 0.000 description 1
- GIVATXIGCXFQQA-FXQIFTODSA-N Arg-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCN=C(N)N GIVATXIGCXFQQA-FXQIFTODSA-N 0.000 description 1
- JTKLCCFLSLCCST-SZMVWBNQSA-N Arg-Arg-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCN=C(N)N)N)C(O)=O)=CNC2=C1 JTKLCCFLSLCCST-SZMVWBNQSA-N 0.000 description 1
- NTAZNGWBXRVEDJ-FXQIFTODSA-N Arg-Asp-Asp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O NTAZNGWBXRVEDJ-FXQIFTODSA-N 0.000 description 1
- FBLMOFHNVQBKRR-IHRRRGAJSA-N Arg-Asp-Tyr Chemical compound NC(N)=NCCC[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 FBLMOFHNVQBKRR-IHRRRGAJSA-N 0.000 description 1
- YUGFLWBWAJFGKY-BQBZGAKWSA-N Arg-Cys-Gly Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CS)C(=O)NCC(O)=O YUGFLWBWAJFGKY-BQBZGAKWSA-N 0.000 description 1
- QAODJPUKWNNNRP-DCAQKATOSA-N Arg-Glu-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O QAODJPUKWNNNRP-DCAQKATOSA-N 0.000 description 1
- DJAIOAKQIOGULM-DCAQKATOSA-N Arg-Glu-Met Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O DJAIOAKQIOGULM-DCAQKATOSA-N 0.000 description 1
- WVNFNPGXYADPPO-BQBZGAKWSA-N Arg-Gly-Ser Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O WVNFNPGXYADPPO-BQBZGAKWSA-N 0.000 description 1
- KRQSPVKUISQQFS-FJXKBIBVSA-N Arg-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCN=C(N)N KRQSPVKUISQQFS-FJXKBIBVSA-N 0.000 description 1
- INXWADWANGLMPJ-JYJNAYRXSA-N Arg-Phe-Arg Chemical compound NC(=N)NCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)CC1=CC=CC=C1 INXWADWANGLMPJ-JYJNAYRXSA-N 0.000 description 1
- OVQJAKFLFTZDNC-GUBZILKMSA-N Arg-Pro-Asp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O OVQJAKFLFTZDNC-GUBZILKMSA-N 0.000 description 1
- JJIBHAOBNIFUEL-SRVKXCTJSA-N Arg-Pro-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CCCN=C(N)N)N JJIBHAOBNIFUEL-SRVKXCTJSA-N 0.000 description 1
- JQHASVQBAKRJKD-GUBZILKMSA-N Arg-Ser-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCCN=C(N)N)N JQHASVQBAKRJKD-GUBZILKMSA-N 0.000 description 1
- LOVIQNMIPQVIGT-BVSLBCMMSA-N Arg-Trp-Phe Chemical compound C([C@H](NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CCCN=C(N)N)N)C(O)=O)C1=CC=CC=C1 LOVIQNMIPQVIGT-BVSLBCMMSA-N 0.000 description 1
- BFDDUDQCPJWQRQ-IHRRRGAJSA-N Arg-Tyr-Cys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N)O BFDDUDQCPJWQRQ-IHRRRGAJSA-N 0.000 description 1
- SWLOHUMCUDRTCL-ZLUOBGJFSA-N Asn-Ala-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N SWLOHUMCUDRTCL-ZLUOBGJFSA-N 0.000 description 1
- RZVVKNIACROXRM-ZLUOBGJFSA-N Asn-Ala-Asp Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CC(=O)N)N RZVVKNIACROXRM-ZLUOBGJFSA-N 0.000 description 1
- NTXNUXPCNRDMAF-WFBYXXMGSA-N Asn-Ala-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CC(N)=O)C)C(O)=O)=CNC2=C1 NTXNUXPCNRDMAF-WFBYXXMGSA-N 0.000 description 1
- MFFOYNGMOYFPBD-DCAQKATOSA-N Asn-Arg-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O MFFOYNGMOYFPBD-DCAQKATOSA-N 0.000 description 1
- HYQYLOSCICEYTR-YUMQZZPRSA-N Asn-Gly-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O HYQYLOSCICEYTR-YUMQZZPRSA-N 0.000 description 1
- LVHMEJJWEXBMKK-GMOBBJLQSA-N Asn-Ile-Met Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CC(=O)N)N LVHMEJJWEXBMKK-GMOBBJLQSA-N 0.000 description 1
- NLRJGXZWTKXRHP-DCAQKATOSA-N Asn-Leu-Arg Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NLRJGXZWTKXRHP-DCAQKATOSA-N 0.000 description 1
- GLWFAWNYGWBMOC-SRVKXCTJSA-N Asn-Leu-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O GLWFAWNYGWBMOC-SRVKXCTJSA-N 0.000 description 1
- MYVBTYXSWILFCG-BQBZGAKWSA-N Asn-Met-Gly Chemical compound CSCC[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CC(=O)N)N MYVBTYXSWILFCG-BQBZGAKWSA-N 0.000 description 1
- YRTOMUMWSTUQAX-FXQIFTODSA-N Asn-Pro-Asp Chemical compound NC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O YRTOMUMWSTUQAX-FXQIFTODSA-N 0.000 description 1
- JBDLMLZNDRLDIX-HJGDQZAQSA-N Asn-Thr-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O JBDLMLZNDRLDIX-HJGDQZAQSA-N 0.000 description 1
- DAYDURRBMDCCFL-AAEUAGOBSA-N Asn-Trp-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CC(=O)N)N DAYDURRBMDCCFL-AAEUAGOBSA-N 0.000 description 1
- CGYKCTPUGXFPMG-IHPCNDPISA-N Asn-Tyr-Trp Chemical compound [H]N[C@@H](CC(N)=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 CGYKCTPUGXFPMG-IHPCNDPISA-N 0.000 description 1
- ZAESWDKAMDVHLL-RCOVLWMOSA-N Asn-Val-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O ZAESWDKAMDVHLL-RCOVLWMOSA-N 0.000 description 1
- UQBGYPFHWFZMCD-ZLUOBGJFSA-N Asp-Asn-Asn Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O UQBGYPFHWFZMCD-ZLUOBGJFSA-N 0.000 description 1
- UGKZHCBLMLSANF-CIUDSAMLSA-N Asp-Asn-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O UGKZHCBLMLSANF-CIUDSAMLSA-N 0.000 description 1
- QXHVOUSPVAWEMX-ZLUOBGJFSA-N Asp-Asp-Ser Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O QXHVOUSPVAWEMX-ZLUOBGJFSA-N 0.000 description 1
- GPPIDDWYKJPRES-YDHLFZDLSA-N Asp-Phe-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O GPPIDDWYKJPRES-YDHLFZDLSA-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
- JDDYEZGPYBBPBN-JRQIVUDYSA-N Asp-Thr-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JDDYEZGPYBBPBN-JRQIVUDYSA-N 0.000 description 1
- USENATHVGFXRNO-SRVKXCTJSA-N Asp-Tyr-Asp Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(O)=O)C(O)=O)CC1=CC=C(O)C=C1 USENATHVGFXRNO-SRVKXCTJSA-N 0.000 description 1
- 238000011725 BALB/c mouse Methods 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 241000252506 Characiformes Species 0.000 description 1
- 102000019034 Chemokines Human genes 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- 241000251730 Chondrichthyes Species 0.000 description 1
- 108010047041 Complementarity Determining Regions Proteins 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- HRJLVSQKBLZHSR-ZLUOBGJFSA-N Cys-Asn-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(O)=O HRJLVSQKBLZHSR-ZLUOBGJFSA-N 0.000 description 1
- NDUSUIGBMZCOIL-ZKWXMUAHSA-N Cys-Asn-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CS)N NDUSUIGBMZCOIL-ZKWXMUAHSA-N 0.000 description 1
- VFGADOJXRLWTBU-JBDRJPRFSA-N Cys-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CS)N VFGADOJXRLWTBU-JBDRJPRFSA-N 0.000 description 1
- AFYGNOJUTMXQIG-FXQIFTODSA-N Cys-Met-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)N AFYGNOJUTMXQIG-FXQIFTODSA-N 0.000 description 1
- FCXJJTRGVAZDER-FXQIFTODSA-N Cys-Val-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O FCXJJTRGVAZDER-FXQIFTODSA-N 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 101100015729 Drosophila melanogaster drk gene Proteins 0.000 description 1
- 102100037024 E3 ubiquitin-protein ligase XIAP Human genes 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- LVGKNOAMLMIIKO-UHFFFAOYSA-N Elaidinsaeure-aethylester Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC LVGKNOAMLMIIKO-UHFFFAOYSA-N 0.000 description 1
- 108091006057 GST-tagged proteins Proteins 0.000 description 1
- GYHNNYVSQQEPJS-OIOBTWANSA-N Gallium-67 Chemical compound [67Ga] GYHNNYVSQQEPJS-OIOBTWANSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- JESJDAAGXULQOP-CIUDSAMLSA-N Gln-Arg-Ser Chemical compound C(C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCC(=O)N)N)CN=C(N)N JESJDAAGXULQOP-CIUDSAMLSA-N 0.000 description 1
- HVQCEQTUSWWFOS-WDSKDSINSA-N Gln-Gly-Cys Chemical compound C(CC(=O)N)[C@@H](C(=O)NCC(=O)N[C@@H](CS)C(=O)O)N HVQCEQTUSWWFOS-WDSKDSINSA-N 0.000 description 1
- HWEINOMSWQSJDC-SRVKXCTJSA-N Gln-Leu-Arg Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O HWEINOMSWQSJDC-SRVKXCTJSA-N 0.000 description 1
- 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 1
- WHVLABLIJYGVEK-QEWYBTABSA-N Gln-Phe-Ile Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WHVLABLIJYGVEK-QEWYBTABSA-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
- SYZZMPFLOLSMHL-XHNCKOQMSA-N Gln-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)N)N)C(=O)O SYZZMPFLOLSMHL-XHNCKOQMSA-N 0.000 description 1
- JILRMFFFCHUUTJ-ACZMJKKPSA-N Gln-Ser-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O JILRMFFFCHUUTJ-ACZMJKKPSA-N 0.000 description 1
- ININBLZFFVOQIO-JHEQGTHGSA-N Gln-Thr-Gly Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CCC(=O)N)N)O ININBLZFFVOQIO-JHEQGTHGSA-N 0.000 description 1
- STHSGOZLFLFGSS-SUSMZKCASA-N Gln-Thr-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O STHSGOZLFLFGSS-SUSMZKCASA-N 0.000 description 1
- HGBHRZBXOOHRDH-JBACZVJFSA-N Gln-Tyr-Trp Chemical compound [H]N[C@@H](CCC(N)=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 HGBHRZBXOOHRDH-JBACZVJFSA-N 0.000 description 1
- QGWXAMDECCKGRU-XVKPBYJWSA-N Gln-Val-Gly Chemical compound CC(C)[C@H](NC(=O)[C@@H](N)CCC(N)=O)C(=O)NCC(O)=O QGWXAMDECCKGRU-XVKPBYJWSA-N 0.000 description 1
- FYBSCGZLICNOBA-XQXXSGGOSA-N Glu-Ala-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O FYBSCGZLICNOBA-XQXXSGGOSA-N 0.000 description 1
- CUXJIASLBRJOFV-LAEOZQHASA-N Glu-Gly-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O CUXJIASLBRJOFV-LAEOZQHASA-N 0.000 description 1
- HILMIYALTUQTRC-XVKPBYJWSA-N Glu-Gly-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O HILMIYALTUQTRC-XVKPBYJWSA-N 0.000 description 1
- SJJHXJDSNQJMMW-SRVKXCTJSA-N Glu-Lys-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O SJJHXJDSNQJMMW-SRVKXCTJSA-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
- DXVOKNVIKORTHQ-GUBZILKMSA-N Glu-Pro-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O DXVOKNVIKORTHQ-GUBZILKMSA-N 0.000 description 1
- 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 1
- VJVAQZYGLMJPTK-QEJZJMRPSA-N Glu-Trp-Asp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CCC(=O)O)N VJVAQZYGLMJPTK-QEJZJMRPSA-N 0.000 description 1
- HHSKZJZWQFPSKN-AVGNSLFASA-N Glu-Tyr-Asp Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(O)=O)C(O)=O HHSKZJZWQFPSKN-AVGNSLFASA-N 0.000 description 1
- BKMOHWJHXQLFEX-IRIUXVKKSA-N Glu-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](CCC(=O)O)N)O BKMOHWJHXQLFEX-IRIUXVKKSA-N 0.000 description 1
- LSYFGBRDBIQYAQ-FHWLQOOXSA-N Glu-Tyr-Tyr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O LSYFGBRDBIQYAQ-FHWLQOOXSA-N 0.000 description 1
- KIEICAOUSNYOLM-NRPADANISA-N Glu-Val-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O KIEICAOUSNYOLM-NRPADANISA-N 0.000 description 1
- VIPDPMHGICREIS-GVXVVHGQSA-N Glu-Val-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O VIPDPMHGICREIS-GVXVVHGQSA-N 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- PYTZFYUXZZHOAD-WHFBIAKZSA-N Gly-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)CN PYTZFYUXZZHOAD-WHFBIAKZSA-N 0.000 description 1
- MZZSCEANQDPJER-ONGXEEELSA-N Gly-Ala-Phe Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 MZZSCEANQDPJER-ONGXEEELSA-N 0.000 description 1
- LJPIRKICOISLKN-WHFBIAKZSA-N Gly-Ala-Ser Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O LJPIRKICOISLKN-WHFBIAKZSA-N 0.000 description 1
- KKBWDNZXYLGJEY-UHFFFAOYSA-N Gly-Arg-Pro Natural products NCC(=O)NC(CCNC(=N)N)C(=O)N1CCCC1C(=O)O KKBWDNZXYLGJEY-UHFFFAOYSA-N 0.000 description 1
- XUORRGAFUQIMLC-STQMWFEESA-N Gly-Arg-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)CN)O XUORRGAFUQIMLC-STQMWFEESA-N 0.000 description 1
- XBWMTPAIUQIWKA-BYULHYEWSA-N Gly-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CN XBWMTPAIUQIWKA-BYULHYEWSA-N 0.000 description 1
- PMNHJLASAAWELO-FOHZUACHSA-N Gly-Asp-Thr Chemical compound [H]NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PMNHJLASAAWELO-FOHZUACHSA-N 0.000 description 1
- KMSGYZQRXPUKGI-BYPYZUCNSA-N Gly-Gly-Asn Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CC(N)=O KMSGYZQRXPUKGI-BYPYZUCNSA-N 0.000 description 1
- GDOZQTNZPCUARW-YFKPBYRVSA-N Gly-Gly-Glu Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O GDOZQTNZPCUARW-YFKPBYRVSA-N 0.000 description 1
- XMPXVJIDADUOQB-RCOVLWMOSA-N Gly-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C([O-])=O)NC(=O)CNC(=O)C[NH3+] XMPXVJIDADUOQB-RCOVLWMOSA-N 0.000 description 1
- UPADCCSMVOQAGF-LBPRGKRZSA-N Gly-Gly-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)CNC(=O)CN)C(O)=O)=CNC2=C1 UPADCCSMVOQAGF-LBPRGKRZSA-N 0.000 description 1
- OLPPXYMMIARYAL-QMMMGPOBSA-N Gly-Gly-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)CNC(=O)CN OLPPXYMMIARYAL-QMMMGPOBSA-N 0.000 description 1
- HMHRTKOWRUPPNU-RCOVLWMOSA-N Gly-Ile-Gly Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O HMHRTKOWRUPPNU-RCOVLWMOSA-N 0.000 description 1
- MHXKHKWHPNETGG-QWRGUYRKSA-N Gly-Lys-Leu Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O MHXKHKWHPNETGG-QWRGUYRKSA-N 0.000 description 1
- PCPOYRCAHPJXII-UWVGGRQHSA-N Gly-Lys-Met Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCSC)C(O)=O PCPOYRCAHPJXII-UWVGGRQHSA-N 0.000 description 1
- QVDGHDFFYHKJPN-QWRGUYRKSA-N Gly-Phe-Cys Chemical compound NCC(=O)N[C@@H](Cc1ccccc1)C(=O)N[C@@H](CS)C(O)=O QVDGHDFFYHKJPN-QWRGUYRKSA-N 0.000 description 1
- IGOYNRWLWHWAQO-JTQLQIEISA-N Gly-Phe-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 IGOYNRWLWHWAQO-JTQLQIEISA-N 0.000 description 1
- OOCFXNOVSLSHAB-IUCAKERBSA-N Gly-Pro-Pro Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 OOCFXNOVSLSHAB-IUCAKERBSA-N 0.000 description 1
- IRJWAYCXIYUHQE-WHFBIAKZSA-N Gly-Ser-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)CN IRJWAYCXIYUHQE-WHFBIAKZSA-N 0.000 description 1
- ZZWUYQXMIFTIIY-WEDXCCLWSA-N Gly-Thr-Leu Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O ZZWUYQXMIFTIIY-WEDXCCLWSA-N 0.000 description 1
- TVTZEOHWHUVYCG-KYNKHSRBSA-N Gly-Thr-Thr Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O TVTZEOHWHUVYCG-KYNKHSRBSA-N 0.000 description 1
- RCHFYMASWAZQQZ-ZANVPECISA-N Gly-Trp-Ala Chemical compound C1=CC=C2C(C[C@@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)CN)=CNC2=C1 RCHFYMASWAZQQZ-ZANVPECISA-N 0.000 description 1
- GNNJKUYDWFIBTK-QWRGUYRKSA-N Gly-Tyr-Asp Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(O)=O)C(O)=O GNNJKUYDWFIBTK-QWRGUYRKSA-N 0.000 description 1
- PNUFMLXHOLFRLD-KBPBESRZSA-N Gly-Tyr-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=C(O)C=C1 PNUFMLXHOLFRLD-KBPBESRZSA-N 0.000 description 1
- LYZYGGWCBLBDMC-QWHCGFSZSA-N Gly-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)CN)C(=O)O LYZYGGWCBLBDMC-QWHCGFSZSA-N 0.000 description 1
- JYGYNWYVKXENNE-OALUTQOASA-N Gly-Tyr-Trp Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O JYGYNWYVKXENNE-OALUTQOASA-N 0.000 description 1
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 1
- 206010019851 Hepatotoxicity Diseases 0.000 description 1
- HTZKFIYQMHJWSQ-INTQDDNPSA-N His-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CN=CN2)N HTZKFIYQMHJWSQ-INTQDDNPSA-N 0.000 description 1
- AKEDPWJFQULLPE-IUCAKERBSA-N His-Glu-Gly Chemical compound N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O AKEDPWJFQULLPE-IUCAKERBSA-N 0.000 description 1
- FYTCLUIYTYFGPT-YUMQZZPRSA-N His-Gly-Ser Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)NCC(=O)N[C@@H](CO)C(O)=O FYTCLUIYTYFGPT-YUMQZZPRSA-N 0.000 description 1
- YXASFUBDSDAXQD-UWVGGRQHSA-N His-Met-Gly Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCSC)C(=O)NCC(O)=O YXASFUBDSDAXQD-UWVGGRQHSA-N 0.000 description 1
- NBWATNYAUVSAEQ-ZEILLAHLSA-N His-Thr-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N)O NBWATNYAUVSAEQ-ZEILLAHLSA-N 0.000 description 1
- WSAILOWUJZEAGC-DCAQKATOSA-N His-Val-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N WSAILOWUJZEAGC-DCAQKATOSA-N 0.000 description 1
- 241001125931 Hoplias malabaricus Species 0.000 description 1
- 229920002153 Hydroxypropyl cellulose Polymers 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- ZDNORQNHCJUVOV-KBIXCLLPSA-N Ile-Gln-Ala Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O ZDNORQNHCJUVOV-KBIXCLLPSA-N 0.000 description 1
- MTFVYKQRLXYAQN-LAEOZQHASA-N Ile-Glu-Gly Chemical compound [H]N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O MTFVYKQRLXYAQN-LAEOZQHASA-N 0.000 description 1
- DFFTXLCCDFYRKD-MBLNEYKQSA-N Ile-Gly-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)O)N DFFTXLCCDFYRKD-MBLNEYKQSA-N 0.000 description 1
- UAQSZXGJGLHMNV-XEGUGMAKSA-N Ile-Gly-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N UAQSZXGJGLHMNV-XEGUGMAKSA-N 0.000 description 1
- PFPUFNLHBXKPHY-HTFCKZLJSA-N Ile-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)O)N PFPUFNLHBXKPHY-HTFCKZLJSA-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
- XMYURPUVJSKTMC-KBIXCLLPSA-N Ile-Ser-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N XMYURPUVJSKTMC-KBIXCLLPSA-N 0.000 description 1
- SAEWJTCJQVZQNZ-IUKAMOBKSA-N Ile-Thr-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N SAEWJTCJQVZQNZ-IUKAMOBKSA-N 0.000 description 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- ZCYVEMRRCGMTRW-AHCXROLUSA-N Iodine-123 Chemical compound [123I] ZCYVEMRRCGMTRW-AHCXROLUSA-N 0.000 description 1
- IBMVEYRWAWIOTN-UHFFFAOYSA-N L-Leucyl-L-Arginyl-L-Proline Natural products CC(C)CC(N)C(=O)NC(CCCN=C(N)N)C(=O)N1CCCC1C(O)=O IBMVEYRWAWIOTN-UHFFFAOYSA-N 0.000 description 1
- UGTHTQWIQKEDEH-BQBZGAKWSA-N L-alanyl-L-prolylglycine zwitterion Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O UGTHTQWIQKEDEH-BQBZGAKWSA-N 0.000 description 1
- LHSGPCFBGJHPCY-UHFFFAOYSA-N L-leucine-L-tyrosine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 LHSGPCFBGJHPCY-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- WGNOPSQMIQERPK-UHFFFAOYSA-N Leu-Asn-Pro Natural products CC(C)CC(N)C(=O)NC(CC(=O)N)C(=O)N1CCCC1C(=O)O WGNOPSQMIQERPK-UHFFFAOYSA-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
- 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 1
- VWHGTYCRDRBSFI-ZETCQYMHSA-N Leu-Gly-Gly Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)NCC(O)=O VWHGTYCRDRBSFI-ZETCQYMHSA-N 0.000 description 1
- HYMLKESRWLZDBR-WEDXCCLWSA-N Leu-Gly-Thr Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O HYMLKESRWLZDBR-WEDXCCLWSA-N 0.000 description 1
- VZBIUJURDLFFOE-IHRRRGAJSA-N Leu-His-Arg Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O VZBIUJURDLFFOE-IHRRRGAJSA-N 0.000 description 1
- HRTRLSRYZZKPCO-BJDJZHNGSA-N Leu-Ile-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O HRTRLSRYZZKPCO-BJDJZHNGSA-N 0.000 description 1
- XVZCXCTYGHPNEM-UHFFFAOYSA-N Leu-Leu-Pro Natural products CC(C)CC(N)C(=O)NC(CC(C)C)C(=O)N1CCCC1C(O)=O XVZCXCTYGHPNEM-UHFFFAOYSA-N 0.000 description 1
- XWEVVRRSIOBJOO-SRVKXCTJSA-N Leu-Pro-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(O)=O XWEVVRRSIOBJOO-SRVKXCTJSA-N 0.000 description 1
- IRMLZWSRWSGTOP-CIUDSAMLSA-N Leu-Ser-Ala Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O IRMLZWSRWSGTOP-CIUDSAMLSA-N 0.000 description 1
- 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 1
- 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 1
- WBRJVRXEGQIDRK-XIRDDKMYSA-N Leu-Trp-Ser Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](N)CC(C)C)C(=O)N[C@@H](CO)C(O)=O)=CNC2=C1 WBRJVRXEGQIDRK-XIRDDKMYSA-N 0.000 description 1
- RIHIGSWBLHSGLV-CQDKDKBSSA-N Leu-Tyr-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(O)=O RIHIGSWBLHSGLV-CQDKDKBSSA-N 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- 239000006142 Luria-Bertani Agar Substances 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
- PAMDBWYMLWOELY-SDDRHHMPSA-N Lys-Glu-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CCCCN)N)C(=O)O PAMDBWYMLWOELY-SDDRHHMPSA-N 0.000 description 1
- WGILOYIKJVQUPT-DCAQKATOSA-N Lys-Pro-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O WGILOYIKJVQUPT-DCAQKATOSA-N 0.000 description 1
- HKXSZKJMDBHOTG-CIUDSAMLSA-N Lys-Ser-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CCCCN HKXSZKJMDBHOTG-CIUDSAMLSA-N 0.000 description 1
- JMNRXRPBHFGXQX-GUBZILKMSA-N Lys-Ser-Glu Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O JMNRXRPBHFGXQX-GUBZILKMSA-N 0.000 description 1
- USPJSTBDIGJPFK-PMVMPFDFSA-N Lys-Tyr-Trp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O USPJSTBDIGJPFK-PMVMPFDFSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 238000000585 Mann–Whitney U test Methods 0.000 description 1
- FRWZTWWOORIIBA-FXQIFTODSA-N Met-Asn-Asn Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N FRWZTWWOORIIBA-FXQIFTODSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- MSFSPUZXLOGKHJ-UHFFFAOYSA-N Muraminsaeure Natural products OC(=O)C(C)OC1C(N)C(O)OC(CO)C1O MSFSPUZXLOGKHJ-UHFFFAOYSA-N 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- HSHXDCVZWHOWCS-UHFFFAOYSA-N N'-hexadecylthiophene-2-carbohydrazide Chemical compound CCCCCCCCCCCCCCCCNNC(=O)c1cccs1 HSHXDCVZWHOWCS-UHFFFAOYSA-N 0.000 description 1
- 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 1
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 1
- 108010079364 N-glycylalanine Proteins 0.000 description 1
- 108010002311 N-glycylglutamic acid Proteins 0.000 description 1
- 241000080590 Niso Species 0.000 description 1
- 108010067902 Peptide Library Proteins 0.000 description 1
- 108010013639 Peptidoglycan 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
- XWBJLKDCHJVKAK-KKUMJFAQSA-N Phe-Arg-Gln Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N XWBJLKDCHJVKAK-KKUMJFAQSA-N 0.000 description 1
- ZENDEDYRYVHBEG-SRVKXCTJSA-N Phe-Asp-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 ZENDEDYRYVHBEG-SRVKXCTJSA-N 0.000 description 1
- CUMXHKAOHNWRFQ-BZSNNMDCSA-N Phe-Asp-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 CUMXHKAOHNWRFQ-BZSNNMDCSA-N 0.000 description 1
- HOYQLNNGMHXZDW-KKUMJFAQSA-N Phe-Glu-Arg Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O HOYQLNNGMHXZDW-KKUMJFAQSA-N 0.000 description 1
- MPFGIYLYWUCSJG-AVGNSLFASA-N Phe-Glu-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 MPFGIYLYWUCSJG-AVGNSLFASA-N 0.000 description 1
- UAMFZRNCIFFMLE-FHWLQOOXSA-N Phe-Glu-Tyr Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N UAMFZRNCIFFMLE-FHWLQOOXSA-N 0.000 description 1
- QPVFUAUFEBPIPT-CDMKHQONSA-N Phe-Gly-Thr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O QPVFUAUFEBPIPT-CDMKHQONSA-N 0.000 description 1
- IEOHQGFKHXUALJ-JYJNAYRXSA-N Phe-Met-Arg Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O IEOHQGFKHXUALJ-JYJNAYRXSA-N 0.000 description 1
- WEDZFLRYSIDIRX-IHRRRGAJSA-N Phe-Ser-Arg Chemical compound NC(=N)NCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CC=CC=C1 WEDZFLRYSIDIRX-IHRRRGAJSA-N 0.000 description 1
- XDMMOISUAHXXFD-SRVKXCTJSA-N Phe-Ser-Asp Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O XDMMOISUAHXXFD-SRVKXCTJSA-N 0.000 description 1
- BONHGTUEEPIMPM-AVGNSLFASA-N Phe-Ser-Glu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O BONHGTUEEPIMPM-AVGNSLFASA-N 0.000 description 1
- 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 1
- ILGCZYGFYQLSDZ-KKUMJFAQSA-N Phe-Ser-His Chemical compound N[C@@H](Cc1ccccc1)C(=O)N[C@@H](CO)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O ILGCZYGFYQLSDZ-KKUMJFAQSA-N 0.000 description 1
- GKRCCTYAGQPMMP-IHRRRGAJSA-N Phe-Ser-Met Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCSC)C(O)=O GKRCCTYAGQPMMP-IHRRRGAJSA-N 0.000 description 1
- MCIXMYKSPQUMJG-SRVKXCTJSA-N Phe-Ser-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O MCIXMYKSPQUMJG-SRVKXCTJSA-N 0.000 description 1
- BPIMVBKDLSBKIJ-FCLVOEFKSA-N Phe-Thr-Phe Chemical compound C([C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 BPIMVBKDLSBKIJ-FCLVOEFKSA-N 0.000 description 1
- MHNBYYFXWDUGBW-RPTUDFQQSA-N Phe-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=CC=CC=C2)N)O MHNBYYFXWDUGBW-RPTUDFQQSA-N 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 108700011066 PreScission Protease Proteins 0.000 description 1
- FUVBEZJCRMHWEM-FXQIFTODSA-N Pro-Asn-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O FUVBEZJCRMHWEM-FXQIFTODSA-N 0.000 description 1
- ILMLVTGTUJPQFP-FXQIFTODSA-N Pro-Asp-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O ILMLVTGTUJPQFP-FXQIFTODSA-N 0.000 description 1
- DXTOOBDIIAJZBJ-BQBZGAKWSA-N Pro-Gly-Ser Chemical compound [H]N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CO)C(O)=O DXTOOBDIIAJZBJ-BQBZGAKWSA-N 0.000 description 1
- BLJMJZOMZRCESA-GUBZILKMSA-N Pro-Met-Asn Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@@H]1CCCN1 BLJMJZOMZRCESA-GUBZILKMSA-N 0.000 description 1
- WLJYLAQSUSIQNH-GUBZILKMSA-N Pro-Met-Ser Chemical compound CSCC[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@@H]1CCCN1 WLJYLAQSUSIQNH-GUBZILKMSA-N 0.000 description 1
- CZCCVJUUWBMISW-FXQIFTODSA-N Pro-Ser-Cys Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O CZCCVJUUWBMISW-FXQIFTODSA-N 0.000 description 1
- FNGOXVQBBCMFKV-CIUDSAMLSA-N Pro-Ser-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O FNGOXVQBBCMFKV-CIUDSAMLSA-N 0.000 description 1
- KWMZPPWYBVZIER-XGEHTFHBSA-N Pro-Ser-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KWMZPPWYBVZIER-XGEHTFHBSA-N 0.000 description 1
- MDAWMJUZHBQTBO-XGEHTFHBSA-N Pro-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@@H]1CCCN1)O MDAWMJUZHBQTBO-XGEHTFHBSA-N 0.000 description 1
- LZHHZYDPMZEMRX-STQMWFEESA-N Pro-Tyr-Gly Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O LZHHZYDPMZEMRX-STQMWFEESA-N 0.000 description 1
- DYJTXTCEXMCPBF-UFYCRDLUSA-N Pro-Tyr-Phe Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)N[C@@H](CC3=CC=CC=C3)C(=O)O DYJTXTCEXMCPBF-UFYCRDLUSA-N 0.000 description 1
- QDDJNKWPTJHROJ-UFYCRDLUSA-N Pro-Tyr-Tyr Chemical compound C([C@@H](C(=O)O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H]1NCCC1)C1=CC=C(O)C=C1 QDDJNKWPTJHROJ-UFYCRDLUSA-N 0.000 description 1
- WWXNZNWZNZPDIF-SRVKXCTJSA-N Pro-Val-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 WWXNZNWZNZPDIF-SRVKXCTJSA-N 0.000 description 1
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 1
- 108010025216 RVF peptide Proteins 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 102000014400 SH2 domains Human genes 0.000 description 1
- 108050003452 SH2 domains Proteins 0.000 description 1
- BTKUIVBNGBFTTP-WHFBIAKZSA-N Ser-Ala-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)NCC(O)=O BTKUIVBNGBFTTP-WHFBIAKZSA-N 0.000 description 1
- WTWGOQRNRFHFQD-JBDRJPRFSA-N Ser-Ala-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WTWGOQRNRFHFQD-JBDRJPRFSA-N 0.000 description 1
- YQHZVYJAGWMHES-ZLUOBGJFSA-N Ser-Ala-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YQHZVYJAGWMHES-ZLUOBGJFSA-N 0.000 description 1
- QGMLKFGTGXWAHF-IHRRRGAJSA-N Ser-Arg-Phe Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O QGMLKFGTGXWAHF-IHRRRGAJSA-N 0.000 description 1
- QVOGDCQNGLBNCR-FXQIFTODSA-N Ser-Arg-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O QVOGDCQNGLBNCR-FXQIFTODSA-N 0.000 description 1
- HBOABDXGTMMDSE-GUBZILKMSA-N Ser-Arg-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(O)=O HBOABDXGTMMDSE-GUBZILKMSA-N 0.000 description 1
- TYYBJUYSTWJHGO-ZKWXMUAHSA-N Ser-Asn-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O TYYBJUYSTWJHGO-ZKWXMUAHSA-N 0.000 description 1
- BTPAWKABYQMKKN-LKXGYXEUSA-N Ser-Asp-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O BTPAWKABYQMKKN-LKXGYXEUSA-N 0.000 description 1
- CDVFZMOFNJPUDD-ACZMJKKPSA-N Ser-Gln-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O CDVFZMOFNJPUDD-ACZMJKKPSA-N 0.000 description 1
- 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 1
- IFPBAGJBHSNYPR-ZKWXMUAHSA-N Ser-Ile-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O IFPBAGJBHSNYPR-ZKWXMUAHSA-N 0.000 description 1
- NNFMANHDYSVNIO-DCAQKATOSA-N Ser-Lys-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NNFMANHDYSVNIO-DCAQKATOSA-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
- SRKMDKACHDVPMD-SRVKXCTJSA-N Ser-Lys-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)N SRKMDKACHDVPMD-SRVKXCTJSA-N 0.000 description 1
- KQNDIKOYWZTZIX-FXQIFTODSA-N Ser-Ser-Arg Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCNC(N)=N KQNDIKOYWZTZIX-FXQIFTODSA-N 0.000 description 1
- 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 1
- NVNPWELENFJOHH-CIUDSAMLSA-N Ser-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CO)N NVNPWELENFJOHH-CIUDSAMLSA-N 0.000 description 1
- XQJCEKXQUJQNNK-ZLUOBGJFSA-N Ser-Ser-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O XQJCEKXQUJQNNK-ZLUOBGJFSA-N 0.000 description 1
- SQHKXWODKJDZRC-LKXGYXEUSA-N Ser-Thr-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O SQHKXWODKJDZRC-LKXGYXEUSA-N 0.000 description 1
- VLMIUSLQONKLDV-HEIBUPTGSA-N Ser-Thr-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VLMIUSLQONKLDV-HEIBUPTGSA-N 0.000 description 1
- PIQRHJQWEPWFJG-UWJYBYFXSA-N Ser-Tyr-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(O)=O PIQRHJQWEPWFJG-UWJYBYFXSA-N 0.000 description 1
- UKKROEYWYIHWBD-ZKWXMUAHSA-N Ser-Val-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O UKKROEYWYIHWBD-ZKWXMUAHSA-N 0.000 description 1
- JGUWRQWULDWNCM-FXQIFTODSA-N Ser-Val-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O JGUWRQWULDWNCM-FXQIFTODSA-N 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 239000012505 Superdex™ Substances 0.000 description 1
- 102000002259 TNF-Related Apoptosis-Inducing Ligand Receptors Human genes 0.000 description 1
- 108010000449 TNF-Related Apoptosis-Inducing Ligand Receptors Proteins 0.000 description 1
- IGROJMCBGRFRGI-YTLHQDLWSA-N Thr-Ala-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O IGROJMCBGRFRGI-YTLHQDLWSA-N 0.000 description 1
- BSNZTJXVDOINSR-JXUBOQSCSA-N Thr-Ala-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O BSNZTJXVDOINSR-JXUBOQSCSA-N 0.000 description 1
- GFDUZZACIWNMPE-KZVJFYERSA-N Thr-Ala-Met Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCSC)C(O)=O GFDUZZACIWNMPE-KZVJFYERSA-N 0.000 description 1
- DGDCHPCRMWEOJR-FQPOAREZSA-N Thr-Ala-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 DGDCHPCRMWEOJR-FQPOAREZSA-N 0.000 description 1
- UNURFMVMXLENAZ-KJEVXHAQSA-N Thr-Arg-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O UNURFMVMXLENAZ-KJEVXHAQSA-N 0.000 description 1
- VIBXMCZWVUOZLA-OLHMAJIHSA-N Thr-Asn-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O VIBXMCZWVUOZLA-OLHMAJIHSA-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
- DCLBXIWHLVEPMQ-JRQIVUDYSA-N Thr-Asp-Tyr Chemical compound C[C@@H](O)[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 DCLBXIWHLVEPMQ-JRQIVUDYSA-N 0.000 description 1
- LYGKYFKSZTUXGZ-ZDLURKLDSA-N Thr-Cys-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)NCC(O)=O LYGKYFKSZTUXGZ-ZDLURKLDSA-N 0.000 description 1
- QQWNRERCGGZOKG-WEDXCCLWSA-N Thr-Gly-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O QQWNRERCGGZOKG-WEDXCCLWSA-N 0.000 description 1
- DJDSEDOKJTZBAR-ZDLURKLDSA-N Thr-Gly-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O DJDSEDOKJTZBAR-ZDLURKLDSA-N 0.000 description 1
- KZSYAEWQMJEGRZ-RHYQMDGZSA-N Thr-Leu-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O KZSYAEWQMJEGRZ-RHYQMDGZSA-N 0.000 description 1
- UJQVSMNQMQHVRY-KZVJFYERSA-N Thr-Met-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C)C(O)=O UJQVSMNQMQHVRY-KZVJFYERSA-N 0.000 description 1
- QHUWWSQZTFLXPQ-FJXKBIBVSA-N Thr-Met-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(=O)NCC(O)=O QHUWWSQZTFLXPQ-FJXKBIBVSA-N 0.000 description 1
- GVMXJJAJLIEASL-ZJDVBMNYSA-N Thr-Pro-Thr Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O GVMXJJAJLIEASL-ZJDVBMNYSA-N 0.000 description 1
- SGAOHNPSEPVAFP-ZDLURKLDSA-N Thr-Ser-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SGAOHNPSEPVAFP-ZDLURKLDSA-N 0.000 description 1
- WPSKTVVMQCXPRO-BWBBJGPYSA-N Thr-Ser-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O WPSKTVVMQCXPRO-BWBBJGPYSA-N 0.000 description 1
- RVMNUBQWPVOUKH-HEIBUPTGSA-N Thr-Ser-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O RVMNUBQWPVOUKH-HEIBUPTGSA-N 0.000 description 1
- GQPQJNMVELPZNQ-GBALPHGKSA-N Thr-Ser-Trp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N)O GQPQJNMVELPZNQ-GBALPHGKSA-N 0.000 description 1
- LECUEEHKUFYOOV-ZJDVBMNYSA-N Thr-Thr-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@@H](N)[C@@H](C)O LECUEEHKUFYOOV-ZJDVBMNYSA-N 0.000 description 1
- NDLHSJWPCXKOGG-VLCNGCBASA-N Thr-Trp-Tyr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CC3=CC=C(C=C3)O)C(=O)O)N)O NDLHSJWPCXKOGG-VLCNGCBASA-N 0.000 description 1
- JAWUQFCGNVEDRN-MEYUZBJRSA-N Thr-Tyr-Leu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC(C)C)C(=O)O)N)O JAWUQFCGNVEDRN-MEYUZBJRSA-N 0.000 description 1
- SJPDTIQHLBQPFO-VLCNGCBASA-N Thr-Tyr-Trp Chemical compound [H]N[C@@H]([C@@H](C)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 SJPDTIQHLBQPFO-VLCNGCBASA-N 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 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 1
- ZWZOCUWOXSDYFZ-CQDKDKBSSA-N Tyr-Ala-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 ZWZOCUWOXSDYFZ-CQDKDKBSSA-N 0.000 description 1
- XHALUUQSNXSPLP-UFYCRDLUSA-N Tyr-Arg-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=C(O)C=C1 XHALUUQSNXSPLP-UFYCRDLUSA-N 0.000 description 1
- NSTPFWRAIDTNGH-BZSNNMDCSA-N Tyr-Asn-Tyr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O NSTPFWRAIDTNGH-BZSNNMDCSA-N 0.000 description 1
- TZXFLDNBYYGLKA-BZSNNMDCSA-N Tyr-Asp-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 TZXFLDNBYYGLKA-BZSNNMDCSA-N 0.000 description 1
- XBWKCYFGRXKWGO-SRVKXCTJSA-N Tyr-Cys-Asn Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(O)=O XBWKCYFGRXKWGO-SRVKXCTJSA-N 0.000 description 1
- RGYDQHBLMMAYNZ-IHRRRGAJSA-N Tyr-Cys-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CC1=CC=C(C=C1)O)N RGYDQHBLMMAYNZ-IHRRRGAJSA-N 0.000 description 1
- GGXUDPQWAWRINY-XEGUGMAKSA-N Tyr-Ile-Gly Chemical compound OC(=O)CNC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 GGXUDPQWAWRINY-XEGUGMAKSA-N 0.000 description 1
- NKUGCYDFQKFVOJ-JYJNAYRXSA-N Tyr-Leu-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 NKUGCYDFQKFVOJ-JYJNAYRXSA-N 0.000 description 1
- KSCVLGXNQXKUAR-JYJNAYRXSA-N Tyr-Leu-Glu 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](CCC(O)=O)C(O)=O KSCVLGXNQXKUAR-JYJNAYRXSA-N 0.000 description 1
- OKDNSNWJEXAMSU-IRXDYDNUSA-N Tyr-Phe-Gly Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)NCC(O)=O)C1=CC=C(O)C=C1 OKDNSNWJEXAMSU-IRXDYDNUSA-N 0.000 description 1
- UUBKSZNKJUJQEJ-JRQIVUDYSA-N Tyr-Thr-Asp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N)O UUBKSZNKJUJQEJ-JRQIVUDYSA-N 0.000 description 1
- NZBSVMQZQMEUHI-WZLNRYEVSA-N Tyr-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](CC1=CC=C(C=C1)O)N NZBSVMQZQMEUHI-WZLNRYEVSA-N 0.000 description 1
- LDKDSFQSEUOCOO-RPTUDFQQSA-N Tyr-Thr-Phe 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](CC1=CC=CC=C1)C(O)=O LDKDSFQSEUOCOO-RPTUDFQQSA-N 0.000 description 1
- GAKBTSMAPGLQFA-JNPHEJMOSA-N Tyr-Thr-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 GAKBTSMAPGLQFA-JNPHEJMOSA-N 0.000 description 1
- QRCBQDPRKMYTMB-IHPCNDPISA-N Tyr-Trp-Ser Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC3=CC=C(C=C3)O)N QRCBQDPRKMYTMB-IHPCNDPISA-N 0.000 description 1
- OJCISMMNNUNNJA-BZSNNMDCSA-N Tyr-Tyr-Asp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C1=CC=C(O)C=C1 OJCISMMNNUNNJA-BZSNNMDCSA-N 0.000 description 1
- AGDDLOQMXUQPDY-BZSNNMDCSA-N Tyr-Tyr-Ser Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(O)=O AGDDLOQMXUQPDY-BZSNNMDCSA-N 0.000 description 1
- DDRBQONWVBDQOY-GUBZILKMSA-N Val-Ala-Arg Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O DDRBQONWVBDQOY-GUBZILKMSA-N 0.000 description 1
- AZSHAZJLOZQYAY-FXQIFTODSA-N Val-Ala-Ser Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O AZSHAZJLOZQYAY-FXQIFTODSA-N 0.000 description 1
- SLLKXDSRVAOREO-KZVJFYERSA-N Val-Ala-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C)NC(=O)[C@H](C(C)C)N)O SLLKXDSRVAOREO-KZVJFYERSA-N 0.000 description 1
- KKHRWGYHBZORMQ-NHCYSSNCSA-N Val-Arg-Glu Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N KKHRWGYHBZORMQ-NHCYSSNCSA-N 0.000 description 1
- QHDXUYOYTPWCSK-RCOVLWMOSA-N Val-Asp-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)NCC(=O)O)N QHDXUYOYTPWCSK-RCOVLWMOSA-N 0.000 description 1
- OVBMCNDKCWAXMZ-NAKRPEOUSA-N Val-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](C(C)C)N OVBMCNDKCWAXMZ-NAKRPEOUSA-N 0.000 description 1
- CXWJFWAZIVWBOS-XQQFMLRXSA-N Val-Lys-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@@H]1C(=O)O)N CXWJFWAZIVWBOS-XQQFMLRXSA-N 0.000 description 1
- VNGKMNPAENRGDC-JYJNAYRXSA-N Val-Phe-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)CC1=CC=CC=C1 VNGKMNPAENRGDC-JYJNAYRXSA-N 0.000 description 1
- XBJKAZATRJBDCU-GUBZILKMSA-N Val-Pro-Ala Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O XBJKAZATRJBDCU-GUBZILKMSA-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
- SVLAAUGFIHSJPK-JYJNAYRXSA-N Val-Trp-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CO)C(=O)O)N SVLAAUGFIHSJPK-JYJNAYRXSA-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
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 1
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 108700031544 X-Linked Inhibitor of Apoptosis Proteins 0.000 description 1
- KRHYYFGTRYWZRS-BJUDXGSMSA-N ac1l2y5h Chemical compound [18FH] KRHYYFGTRYWZRS-BJUDXGSMSA-N 0.000 description 1
- DHKHKXVYLBGOIT-UHFFFAOYSA-N acetaldehyde Diethyl Acetal Natural products CCOC(C)OCC DHKHKXVYLBGOIT-UHFFFAOYSA-N 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 101150063416 add gene Proteins 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 230000001270 agonistic effect Effects 0.000 description 1
- 108010045350 alanyl-tyrosyl-alanine Proteins 0.000 description 1
- 108010044940 alanylglutamine Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 108010070783 alanyltyrosine Proteins 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 239000008365 aqueous carrier Substances 0.000 description 1
- 108010013835 arginine glutamate Proteins 0.000 description 1
- 108010001271 arginyl-glutamyl-arginine Proteins 0.000 description 1
- 108010069926 arginyl-glycyl-serine Proteins 0.000 description 1
- 108010068380 arginylarginine Proteins 0.000 description 1
- 108010062796 arginyllysine Proteins 0.000 description 1
- 108010060035 arginylproline Proteins 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 108010093581 aspartyl-proline Proteins 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 239000012148 binding buffer Substances 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 239000002981 blocking agent Substances 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000003114 blood coagulation factor Substances 0.000 description 1
- 229940019700 blood coagulation factors Drugs 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 229910052792 caesium Inorganic materials 0.000 description 1
- TVFDJXOCXUVLDH-UHFFFAOYSA-N caesium atom Chemical compound [Cs] TVFDJXOCXUVLDH-UHFFFAOYSA-N 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 229920003123 carboxymethyl cellulose sodium Polymers 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 229940105329 carboxymethylcellulose Drugs 0.000 description 1
- 229940084030 carboxymethylcellulose calcium Drugs 0.000 description 1
- 229940063834 carboxymethylcellulose sodium Drugs 0.000 description 1
- 239000004359 castor oil Substances 0.000 description 1
- 235000019438 castor oil Nutrition 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- RYGMFSIKBFXOCR-IGMARMGPSA-N copper-64 Chemical compound [64Cu] RYGMFSIKBFXOCR-IGMARMGPSA-N 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 150000001945 cysteines Chemical class 0.000 description 1
- 108010016616 cysteinylglycine Proteins 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000002059 diagnostic imaging Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- ASXBYYWOLISCLQ-HZYVHMACSA-N dihydrostreptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](CO)(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 ASXBYYWOLISCLQ-HZYVHMACSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 150000002019 disulfides Chemical class 0.000 description 1
- 229940000406 drug candidate Drugs 0.000 description 1
- 239000012149 elution buffer Substances 0.000 description 1
- 230000007515 enzymatic degradation Effects 0.000 description 1
- LVGKNOAMLMIIKO-QXMHVHEDSA-N ethyl oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC LVGKNOAMLMIIKO-QXMHVHEDSA-N 0.000 description 1
- 229940093471 ethyl oleate Drugs 0.000 description 1
- IYBKWXQWKPSYDT-UHFFFAOYSA-L ethylene glycol disuccinate bis(sulfo-N-succinimidyl) ester sodium salt Chemical compound [Na+].[Na+].O=C1C(S(=O)(=O)[O-])CC(=O)N1OC(=O)CCC(=O)OCCOC(=O)CCC(=O)ON1C(=O)C(S([O-])(=O)=O)CC1=O IYBKWXQWKPSYDT-UHFFFAOYSA-L 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 238000001641 gel filtration chromatography Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 108010080575 glutamyl-aspartyl-alanine Proteins 0.000 description 1
- 108010079547 glutamylmethionine Proteins 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 1
- 108010075431 glycyl-alanyl-phenylalanine Proteins 0.000 description 1
- 108010051307 glycyl-glycyl-proline Proteins 0.000 description 1
- 108010010096 glycyl-glycyl-tyrosine Proteins 0.000 description 1
- 108010059898 glycyl-tyrosyl-lysine Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- 108010081551 glycylphenylalanine Proteins 0.000 description 1
- 108010077515 glycylproline Proteins 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 101150098203 grb2 gene Proteins 0.000 description 1
- 230000007686 hepatotoxicity Effects 0.000 description 1
- 231100000304 hepatotoxicity Toxicity 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 150000007857 hydrazones Chemical class 0.000 description 1
- 229920001477 hydrophilic polymer Polymers 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 239000001863 hydroxypropyl cellulose Substances 0.000 description 1
- 235000010977 hydroxypropyl cellulose Nutrition 0.000 description 1
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 1
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 1
- 229940071676 hydroxypropylcellulose Drugs 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000000951 immunodiffusion Effects 0.000 description 1
- APFVFJFRJDLVQX-AHCXROLUSA-N indium-111 Chemical compound [111In] APFVFJFRJDLVQX-AHCXROLUSA-N 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- XMBWDFGMSWQBCA-YPZZEJLDSA-N iodane Chemical compound [125IH] XMBWDFGMSWQBCA-YPZZEJLDSA-N 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 1
- 108010034529 leucyl-lysine Proteins 0.000 description 1
- 108010012058 leucyltyrosine Proteins 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 108010003700 lysyl aspartic acid Proteins 0.000 description 1
- 239000006249 magnetic particle Substances 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 229960001855 mannitol Drugs 0.000 description 1
- 150000004667 medium chain fatty acids Chemical class 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229940125645 monoclonal antibody drug Drugs 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 229910052759 nickel Inorganic materials 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- ZWLPBLYKEWSWPD-UHFFFAOYSA-N o-toluic acid Chemical compound CC1=CC=CC=C1C(O)=O ZWLPBLYKEWSWPD-UHFFFAOYSA-N 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 150000002895 organic esters Chemical class 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 108010084525 phenylalanyl-phenylalanyl-glycine Proteins 0.000 description 1
- 108010084572 phenylalanyl-valine Proteins 0.000 description 1
- 108010012581 phenylalanylglutamate Proteins 0.000 description 1
- 108010083476 phenylalanyltryptophan Proteins 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 108010004914 prolylarginine Proteins 0.000 description 1
- 235000019833 protease Nutrition 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 108010029895 rubimetide Proteins 0.000 description 1
- 239000012146 running buffer Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000003118 sandwich ELISA Methods 0.000 description 1
- 238000007480 sanger sequencing Methods 0.000 description 1
- UQDJGEHQDNVPGU-UHFFFAOYSA-N serine phosphoethanolamine Chemical class [NH3+]CCOP([O-])(=O)OCC([NH3+])C([O-])=O UQDJGEHQDNVPGU-UHFFFAOYSA-N 0.000 description 1
- 108010048818 seryl-histidine Proteins 0.000 description 1
- 108010069117 seryl-lysyl-aspartic acid Proteins 0.000 description 1
- RMAQACBXLXPBSY-UHFFFAOYSA-N silicic acid Chemical compound O[Si](O)(O)O RMAQACBXLXPBSY-UHFFFAOYSA-N 0.000 description 1
- 235000012239 silicon dioxide Nutrition 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 108010005652 splenotritin Proteins 0.000 description 1
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 108010004034 stable plasma protein solution Proteins 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 229940032147 starch Drugs 0.000 description 1
- 238000012916 structural analysis Methods 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 150000005846 sugar alcohols Polymers 0.000 description 1
- 125000004434 sulfur atom Chemical group 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000004797 therapeutic response Effects 0.000 description 1
- 150000003568 thioethers Chemical class 0.000 description 1
- 108010033670 threonyl-aspartyl-tyrosine Proteins 0.000 description 1
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 1
- 108010058119 tryptophyl-glycyl-glycine Proteins 0.000 description 1
- 238000004879 turbidimetry Methods 0.000 description 1
- 108010035534 tyrosyl-leucyl-alanine Proteins 0.000 description 1
- 108010051110 tyrosyl-lysine Proteins 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 229910052720 vanadium Inorganic materials 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 229960001134 von willebrand factor Drugs 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/005—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies constructed by phage libraries
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
-
- 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
- C07K16/36—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against blood coagulation factors
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/46—Hybrid immunoglobulins
- C07K16/468—Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
- C07K2317/14—Specific host cells or culture conditions, e.g. components, pH or temperature
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/22—Immunoglobulins specific features characterized by taxonomic origin from camelids, e.g. camel, llama or dromedary
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
- C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
- C07K2317/35—Valency
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—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
- C07K2317/569—Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Virology (AREA)
- Hematology (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明涉及由D‑氨基酸和非手性甘氨酸构成的低分子抗体及其制造方法,以及利用与靶蛋白对应的镜像型靶蛋白和抗体库的所述低分子抗体的筛选方法。
Description
技术领域
相关申请
本申请要求基于日本专利申请2020-105104(2020年6月18日申请)的优先权,其内容作为参照并入本说明书。
技术领域
本发明涉及免疫原性降低的低分子抗体及其制造方法。更具体地,本发明涉及由D-氨基酸和非手性甘氨酸构成的低分子抗体及其制造方法,以及利用与靶蛋白对应的镜像型靶蛋白和抗体库的所述低分子抗体的筛选。
背景技术
单克隆抗体是治疗领域最具吸引力的模态之一。高亲和力、高选择性和长血半衰期是该模态的主要优点,但其尺寸(~150kDa)和结构的复杂性限制了适用范围。为了解决尺寸和稳定性的问题,已经开发了若干抗体样支架,例如Adnectin、Affibody(亲和体)、Affilin等。来源于这些支架的药物候选分子尽管比以往的抗体小得多,但具有良好的亲和力和稳定性。但是,在这些抗体样支架中,由于抗药物抗体(ADA)的出现导致的效果减弱或意想不到的副作用等,其免疫原性常常成为问题。
免疫原性问题不仅存在于抗体样支架中,也存在于已获批准的单克隆抗体药物中。因此,期望免疫原性的预测以及免疫原性降低的新药物支架的开发。
已知体内的蛋白酶是手性的,能够区分L-及D-蛋白质之间的结构差异,因此D-蛋白质的分解稳定性大幅度提高。报道了将镜像噬菌体展示应用于D-蛋白质支架库,合成具有高亲和力和稳定性的VEGF-A抑制剂。由D-氨基酸构成的该VEGF-A抑制剂不显示免疫原性(非专利文献1和2)。
本发明人等建立了虚拟镜像库的筛选工艺,报道了新的癌症治疗方法的开发(非专利文献3至6)。另外,若干团体报道了噬菌体展示肽库的镜像筛选(上述非专利文献1和2等)。
作为骆驼科动物固有的仅有重链的抗体的单一结构域抗原结合片段的VHH抗体是作为新一代药剂候选的最具吸引力的支架之一(非专利文献7)。尽管其尺寸较小(~15kDa),VHH抗体具有与以往的抗体(~150kDa)同样的高特异性和亲和力。此外,其高稳定性和溶解性适用于药物开发。2019年,第一种基于VHH抗体的药剂被FDA批准,并且关于若干候选分子的临床试验正在进行中。据说VHH抗体免疫原性较低,但详细情况还不清楚。VHH抗体具有重折叠功能,最近还报道了通过化学合成制造VHH抗体(非专利文献8)。然而,对于VHH抗体,也存在因立体结构在免疫细胞中的分解而产生抗药物抗体或中和抗体的风险(非专利文献9和10)。
现有技术文献
非专利文献
非专利文献1:Mandal,K.等人,(2012)Chemical synthesis and X-raystructure of a heterochiral{D-protein antagonist plus vascular endothelialgrowth factor}protein complex by racemic crystallography.Proc.Natl.Acad.Sci.U.S.A.109,14779-14784.
非专利文献2:Uppalapati,M.等人,(2016)A potent D-protein antagonist ofVEGF-A is nonimmunogenic,metabolically stable,and longer-circulating invivo.ACS Chem.Biol.11,1058-1065.
非专利文献3:Noguchi,T.等人,(2016)Screening of a virtual mirror-imagelibrary of natural products.Chem.Commum.52,7653-7656.
非专利文献4:Noguchi,T.等人,(2017)Synthesis of Grb2 SH2 domainproteins for mirror-image screening systems.Bioconjugate Chem.28,609-619.
非专利文献5:Shu,K.等人,(2017)Synthesis of the Src SH2 domain and itsapplication in bioassays for mirror-image screening.RSC Adv.7,38725-38732.
非专利文献6:Shu,K.等人,(2019)Development of mirror-image screeningsystems for XIAP BIR3 domain inhibitors.Bioconjugate Chem.30,1395-1404.
非专利文献7:Muyldermans,S.(2013)Nanobodies:natural single-domainantibodies.Annu.Rev.Biochem.82,775-797.
非专利文献8:Hartmann,L.等人,(2019)VHH characterization.Comparison ofrecombinant with chemically synthesized anti-HER2 VHH.Protein Sci.28,1865-1879.
非专利文献9:Papadopoulos,K.P.,(2015)Unexpected Hepatotoxicity in aPhase I Study of TAS266,a Novel Tetravalent Agonistic Nanobody(R)Targeting theDR5 Receptor.Cancer Chemother Pharmacol.75(5),887-895.
非专利文献10:Kibria M.G.,等人,(2020)The immunogenicity of an anti-EGFR single domain antibody(V(HH))is enhanced by misfolded amorphousaggregation but not by heat-aggregation.Eur J Pharm Biopharm.152,164-174.
发明内容
发明要解决的问题
本发明的课题是提供一种降低了免疫原性的新的药物产品模态。
用于解决课题的手段
已知由作为天然L-氨基酸的对映体的D-氨基酸构成的镜像型的肽·蛋白质难以受到生物体内的蛋白酶的底物识别。发明人等认为,通过将抗体分子镜像化,或许可以在维持其特异性的同时降低免疫原性。
为了确认上述概念,发明人等着眼于作为具有抗体功能的最小结构域而被熟知的VHH抗体,利用自然化学连接(NCL)法,建立了由D-氨基酸和非手性甘氨酸构成的镜像型VHH抗体的化学合成法。因此,对于得到的镜像型VHH抗体,确认了其高结合特异性和免疫原性的降低。
本发明基于上述认知,提供以下[1]至[14]。
[1]一种低分子抗体,所述低分子抗体由D-氨基酸和非手性甘氨酸构成,并且所述低分子抗体的分子量为60KDa及以下、更优选为24KDa及以下、进一步优选为15KDa及以下。
所述低分子抗体优选为500个氨基酸及以下、更优选为200个氨基酸及以下、进一步优选为120个氨基酸及以下,具体地,包括单一结构域抗体、scFv、dsFV、Fab、Fab’等。
[2]根据[1]所述的低分子抗体,所述低分子抗体是选自单一结构域抗体、scFV和Fab中的任一种、优选是单一结构域抗体或scFV、更优选是单一结构域抗体。
单一结构域抗体包括VHH抗体(包括人化VHH抗体)、IgNAR抗体(VNAR抗体)、骆驼化VH抗体等。
[3]根据[1]所述的低分子抗体,所述低分子抗体是VHH抗体。
[4]根据[1]所述的低分子抗体,所述低分子抗体是具有:
(1)由序列号1~4表示的氨基酸序列或由具有与所述序列至少60%、70%、80%或90%的同一性的氨基酸序列分别表示的4个骨架区;
(2)由序列号5~8表示的氨基酸序列或由具有与所述序列至少60%、70%、80%或90%的同一性的氨基酸序列分别表示的4个骨架区;
(3)由序列号9~12表示的氨基酸序列或由具有与所述序列至少60%、70%、80%或90%的同一性的氨基酸序列分别表示的4个骨架区;
(4)由序列号13~16表示的氨基酸序列或由具有与所述序列至少60%、70%、80%或90%的同一性的氨基酸序列分别表示的4个骨架区;或者
(5)由序列号17~20表示的氨基酸序列或由具有与所述序列至少60%、70%、80%或90%的同一性的氨基酸序列分别表示的4个骨架区;的VHH抗体。
[5]一种由D-氨基酸和非手性甘氨酸构成的低分子抗体的制造方法,所述方法包括:
1)将低分子抗体的氨基酸序列分割为以半胱氨酸残基或丙氨酸残基为N末端的多个片段;
2)通过固相法化学合成各片段;以及
3)通过NCL法将合成的各片段依次连结,合成由D-氨基酸和非手性甘氨酸构成的低分子抗体。
[6]根据[5]所述的方法,其中,所述低分子抗体是选自单一结构域抗体、scFV和Fab中的任一种。
[7]根据[5]所述的方法,其中,所述低分子抗体是VHH抗体。
当所述低分子抗体是VHH抗体时,优选片段被FR1和FR3中存在的2个半胱氨酸和其间存在的适当的半胱氨酸或丙氨酸残基分割为4个片段。例如,可以使用上述[4]所述的作为VHH抗体。
[8]一种针对靶蛋白的、由D-氨基酸和非手性甘氨酸构成的低分子抗体的制造方法,所述方法包括:
1)提供针对靶蛋白的、由D-氨基酸和非手性甘氨酸构成的镜像型靶蛋白;
2)使用所述镜像型靶蛋白筛选抗体库,取得对所述镜像型靶蛋白具有亲和力的抗体;以及
3)基于得到的抗体的氨基酸序列,合成由D-氨基酸和非手性甘氨酸构成的低分子抗体。
优选噬菌体展示库等作为库。另外,根据需要,为了提高工序2)中得到的抗体的亲和力,也可以实施亲合力成熟。
[9]根据[8]所述的方法,其中,由D-氨基酸和非手性甘氨酸构成的低分子抗体的合成包括以下工序:
1)将低分子抗体的氨基酸序列分割为以半胱氨酸残基或丙氨酸残基为N末端的多个片段;
2)通过固相法化学合成各片段;以及
3)通过NCL法将合成的各片段依次连结,合成由D-氨基酸和非手性甘氨酸构成的低分子抗体。
[10]根据[8]所述的方法,其中,所述低分子抗体是选自单一结构域抗体、scFV和Fab中的任一种。
[11]根据[8]所述的方法,其中,所述低分子抗体是VHH抗体。例如,可以使用上述[4]所述的作为VHH抗体。
[12]一种多价或多特异性抗体,所述多价或多特异性抗体通过连结[1]至[4]中任一项所述的低分子抗体而得到。
[13]一种由D-氨基酸构成的多价或多特异性抗体的制造方法,所述制造方法包括连结用[5]至[11]中任一项所述的方法制造的低分子抗体。
[14]一种由D-氨基酸构成的多价或多特异性抗体的制造用试剂盒,所述试剂盒包括分别包含下述(a)或(b)的氨基酸序列的、由D-氨基酸和非手性甘氨酸构成的一个或多个多肽:
(a)由序列号55~57、61及63中任一个表示的氨基酸序列;
(b)具有与由序列号55~57、61及63中任一个表示的氨基酸序列(优选在与所述序列的骨架区对应的部分中)至少60%、70%、80%、90%、95%、98%或99%的同一性的氨基酸序列。
优选上述试剂盒包括具有由序列号55或序列号63表示的氨基酸序列或者具有与所述序列(优选在与所述序列的骨架区对应的部分中)至少60%、70%、80%、90%、95%、98%或99%的同一性的氨基酸序列的多肽。
发明的效果
本发明涉及的由D-氨基酸和非手性甘氨酸构成的低分子抗体,在维持与完整长度的抗体同样的特异性和亲和力的同时,免疫原性降低。另外,与完整长度的抗体相比较,稳定性和溶解性高,也适合药物开发。
本发明的低分子抗体,靶蛋白的结构也与由L-氨基酸组成的天然型低分子抗体呈完全镜像的关系。因此,通过使用镜像型靶蛋白筛选抗体库,可以筛选出靶蛋白特异性的、由D-氨基酸和非手性甘氨酸构成的低分子抗体。
附图说明
【图1】虚拟D-抗体库筛选策略概念图。示出了与使用L-蛋白质的D-抗体库的筛选对应的、使用D-蛋白质的L-抗体库的筛选。
【图2】GFP-VHH抗体、抗HER2 VHH抗体和卡拉西单抗(Caplacizumab)的序列(序列号52~54)。红色:半胱氨酸;橙色:丙氨酸;阴影部分:互补性决定区。
【图3】示出了L-GFP-VHH抗体/D-GFP-VHH抗体的合成工艺。(a)Fmoc-Arg(Pbf)-OH、HBTU、HOBt、(i-Pr)2NEt、DMF、之后是20%哌啶/DMF;(b)Fmoc-Dbz-OH、HBTU、HOBt、(i-Pr)2NEt、DMF、之后是20%哌啶/DMF;(c)Fmoc-Xaa-OH、HBTU、HOBt、(i-Pr)2NEt、DMF、之后是20%哌啶/DMF[重复(c)直到序列完结];(d)Boc2O、(i-Pr)2NEt和DMF;(e)4-硝基苯基氯甲酸酯和DCM;(f)(i-Pr)2NEt和DMF;(g)TFA/H2O/m-甲酚/硫代茴香醚/1,2-乙二硫醇(EDT)(80:5:5:5:5);(h)MESNa、6M胍和200mM磷酸缓冲液(pH 6.0);(i)Boc-Cys(Acm)-OH、HBTU、HOBt、(i-Pr)2NEt和DMF;(j)MPAA、100mM TCEP、6M胍和200mM磷酸缓冲液(pH 7.0);(k)20mM VA-044、100mM MESNa、250mM TCEP、6M胍和100mM磷酸缓冲液(pH 6.5);(l)PdCl2、6M胍、100mM磷酸缓冲液、之后是DTT;以及(m)NaNO2、6M胍、200mM磷酸缓冲液、之后是MPAA和TCEP
【图4】通过重组L-GFP-VHH抗体、合成L-GFP-VHH抗体及合成D-GFP-VHH抗体的CD光谱测定的结构分析。(A)CD光谱。(B)热稳定性评价。
【图5】GST-EGFP或GST-mCherry与重组或合成L-GFP-VHH抗体间的夹心ELISA。示出了3次分析的平均吸光度值±SD。
【图6】针对重组L-GFP-VHH抗体、合成L-GFP-VHH抗体及合成D-GFP-VHH抗体的GST-EGFP及GST-mCherry的QCM分析的代表性数据。Langmuir模型被用于确定3次分析的结合亲和力。
【图7】合成L-GFP-VHH抗体及合成D-GFP-VHH抗体的免疫原性。(A)用于各种抗原浓度下ADA测定的ELISA。分析第28天收集的血清。(B)用于对第0、14、21及28天收集的血清的ADA进行测定的ELISA。被包衣的抗原的浓度为1000ng/mL。示出了关于施用合成L-GFP-VHH抗体及合成D-GFP-VHH抗体的小鼠的ADA产生,通过曼-惠特尼U检验算出的统计显著性差异。
【图8】公知的VHH抗体的序列比对。3个CDR区用阴影表示,FR区的突变位点被高亮。如图所示,FR区最多包含16个突变。各序列从上到下依次如序列表的序列号21~51所示。
【图9】示出了PMP12A2h1/D-PMP12A2h1的合成工艺。(a)Fmoc-Arg(Pbf)-OH、HBTU、HOBt、(i-Pr)2NEt、DMF、之后是20%哌啶/DMF;(b)Fmoc-Dbz-OH或Fmoc-MeDbz-OH或Fmoc-(o-Boc)Dbz-OH、HBTU、HOBt、(i-Pr)2NEt、DMF、之后是20%哌啶/DMF;(c)Fmoc-Xaa-OH、Oxyma Pure、DIC、DMF、之后是20%哌啶/DMF[重复(c)直到序列完结];(d)Boc2O、(i-Pr)2NEt和DMF;(e)4-硝基苯基氯甲酸酯和DCM;(f)(i-Pr)2NEt和DMF;(g)TFA/H2O/m-甲酚/硫代茴香醚/1,2-乙二硫醇(EDT)(80:5:5:5:5);(h)Boc-Cys(Acm)-OH、Oxyma Pure、DIC和DMF;(i)MPAA、TCEP、6M胍和200mM磷酸缓冲液(pH 7.0);(j)VA-044、MESNa、TCEP、6M胍和200mM磷酸缓冲液(pH6.5);(k)PdCl2、6M胍、200mM磷酸缓冲液、之后是DTT;以及(l)NaNO2、6M胍、200mM磷酸缓冲液、之后是MPAA和TCEP
【图10】示出了PMP12A2h1(实线)和D-PMP12A2h1(虚线)的CD光谱。
【图11】示出了通过表面等离子共振分析PMP12A2h1与vWF A1结构域的结合亲和力的结果。图示出了当PMP12A2h1自上在400nM、200nM、100nM、50nM、25nM、13nM、6.3nM、3.1nM、1.6nM、0nM处流动时的数据。解离常数KD=39±2nM
具体实施方式
1.本发明的低分子抗体
“低分子抗体”
本说明书中,“低分子抗体”意指具有比完整长度的抗体(IgG的情况下,分子量约150KDa)更低的分子量、但对抗原具有亲和力和特异性的抗体、抗体片段或抗体样分子。
本发明的低分子抗体的靶抗原(靶蛋白)没有特别限定,但考虑到治疗和诊断性应用,优选为识别肿瘤相关抗原、与炎症或过敏相关的抗原、与病毒或细菌感染相关的抗原、或者与循环器官疾病相关的抗原的低分子抗体。
本发明的低分子抗体的分子量优选为60KDa及以下、更优选为24KDa及以下、进一步优选为15KDa及以下。
本发明的低分子抗体的氨基长度优选为500个氨基酸及以下、更优选为200个氨基酸及以下、进一步优选为120个氨基酸及以下。
作为本发明的低分子抗体,例如可以举出单一结构域抗体、scFv、dsFV、Fab、Fab’等。
低分子抗体优选是单一结构域抗体、scFV和Fab、更优选是单一结构域抗体或scFV、进一步优选是单一结构域抗体。如后所述,将这些低分子抗体连结、多聚体化而得到的双抗体(scFV的二聚体)、F(ab')2(Fab’的二聚体)等多价抗体或多特异性抗体也包含在本发明中。在这些多聚体中,构成该多聚体的单体的平均分子量和氨基酸长度优选在上述范围内。
“单一结构域抗体”
单一结构域抗体(SDAB:Single Domain Antibody)是由具有抗原结合性的单一的结构域(单一可变结构域)构成的抗体分子,例如可以举出由骆驼科动物(骆驼、美洲驼、羊驼等)的重链抗体设计的VHH抗体(包括人化VHH抗体)、由软骨鱼类(鲨鱼等)的重链抗体设计的IgNAR抗体(VNAR抗体)、骆驼化VH抗体等。
“VHH抗体”
单一结构域抗体优选是VHH抗体。VHH(Variable domain of Heavy chain ofHeavy chain antibody)抗体是来源于由骆驼科动物仅有的H链构成的重链抗体的可变区的低分子抗体,也被称为纳米抗体(注册商标)。
由于VHH抗体的平均分子量约为15kDa,氨基酸长度约为110~120个氨基酸左右,因此可以在普通的微生物表达系中进行重组生产,如上所述,也报道了通过化学合成的生产。
VHH抗体具有热稳定性及立体结构可逆性高、热变性后重折叠易恢复原立体结构的这一特性。另外,据称与完整长度的抗体相比较,一般来说免疫原性低、安全。
VHH抗体由对于抗原的结合重要的3个CDR区(CDR1、CDR2、CDR3)和包围这些区的更保守的4个骨架区(FR1、FR2、FR3、FR4)构成。
VHH抗体的结构:
FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4
Muyldermans等人报道了根据Kabat等人的EU编号,在骆驼来源的VHH抗体中,FR1包含在第1~30位、CDR1包含在第31~35位、FR2包含在第36~49位、CDR2包含在第50~65位、FR3包含在第66~94位、CDR3包含在第95~102位、FR4包含在第103~113位(LRiechmann和S Muyldermans,J Immunol Methods.1999Dec 10;231(1-2):25-38)。但是,以上这些位置遵循Kabat等人的EU编号,在其他动物来源的VHH抗体中可能不同,也可能与实际的序列中的位置不一致。另外,在IMGT标记或其他编号中有时可能不同。
在一个实施方案中,FR1、FR2、FR3、FR4具有以下序列(实施例中使用的VHH抗体的骨架区)。
FR1:QVQLVESGGALVQPGGSLRLSCAAS(序列号1)
FR2:WYRQAPGKEREWVA(序列号2)
FR3:YEDSVKGRFTISRDDARNTVYLQMNSLKPEDTAVYYCN(序列号3)
FR4:WGQGTQVTVSS(序列号4)
在另一个实施方案中,FR1、FR2、FR3、FR4具有以下序列(美洲驼VHH抗体(sdAb_7047_Lg)的骨架区)。
FR1:QVQLQESGGGLVQAGGSLRLSCAAA(序列号5)
FR2:WFRQAPGKEREFVG(序列号6)
FR3:YADSVKGRFIISRDNAKNTVYLQMNSLKPEDTAVYYCA(序列号7)
FR4:WGQGTQVTVSS(序列号8)
在又另一个实施方案中,FR1、FR2、FR3、FR4具有以下序列(羊驼VHH抗体(sdAb_6474_Vp)的骨架区)。
FR1:QVQLQESGGGLVQPGGSLRLSCAAS(序列号9)
FR2:WYRQAPGKERELVA(序列号10)
FR3:DSVKGRYTISRDYAKNTVYLQMNSLKPEDTALYYCN(序列号11)
FR4:WGQGTQVTVSS(序列号12)
在又另一个实施方案中,FR1、FR2、FR3、FR4具有以下序列(羊驼VHH抗体(sdAb_0039_Vp)的骨架区)。
FR1:QVQLQESGGGLVQPGGSLRLSCAAS(序列号13)
FR2:WFRQAPGKEREFVA(序列号14)
FR3:YADSVKGRFTISRDNAKNTVYLQMNSLKPEDTAVYYCA(序列号15)
FR4:WGQGTQVTVSS(序列号16)
在又另一个实施方案中,FR1、FR2、FR3、FR4具有以下序列(骆驼VHH抗体(sdAb_5835_Cd)的骨架区)。
FR1:QVQLVESGGGSVQAGGSLRLSCTAS(序列号17)
FR2:WYHQAPGNECELVS(序列号18)
FR3:YADSVKGRFTISRDNAKNTVYLQMNSLKPEDTAMYYCA(序列号19)
FR4:WGQGTQVTVSS(序列号20)
上述序列是一个例子,本领域技术人员能够适当地利用图8所述的公知的VHH抗体或数据库(Single Domain Antibody Database:http://www.sdab-db.ca/?)中注册的其他公知的VHH抗体的骨架区的序列。
与CDR区不同,骨架区能够被变化而不会对保守置换等抗体的结合特性产生不利的影响(Mitchell LS.和Colwell LJ.,Proteins(2018)86(7):697-706)。另外,改变对抗体分子和抗原结合部位的稳定化重要的骨架残基,提高抗体的稳定性和特异性的技术也是本领域公知的。
具有对于某一氨基酸序列来说一个或多个氨基酸残基的缺失、附加和/或通过用其它氨基酸置换而修饰的氨基酸序列的多肽维持其生物学活性是本领域公知的(Mark,D.F.等人,Proc.Natl.Acad.Sci.USA(1984)81,5662-5666、Zoller,M.J.和Smith,M.,Nucleic Acids Research(1982)10,6487-6500、Wang,A.等人,Science 224,1431-1433、Dalbadie-McFarland,G.等人,Proc.Natl.Acad.Sci.USA(1982)79,6409-6413)。
例如,基于侧链的性质进行分类时,在构成某一多肽的氨基酸序列中,被分类为同一组的氨基酸即使相互置换,维持该多肽的活性的可能性也很高,这样的组内的氨基酸间的置换称为保守置换。
疏水性氨基酸(A、I、L、M、F、P、W、Y、V)、
亲水性氨基酸(R、D、N、C、E、Q、G、H、K、S、T)、
具有脂肪族侧链的氨基酸(G、A、V、L、I、P)、
具有含羟基侧链的氨基酸(S、T、Y)、
具有含硫原子侧链的氨基酸(C、M)、
具有含羧酸和酰胺侧链的氨基酸(D、N、E、Q)、
具有含碱基侧链的氨基酸(R、K、H)、
具有含芳香族侧链的氨基酸(H、F、Y、W)
因此,只要具有规定的抗原特异性,骨架区可以是具有与由上述序列号1~4、序列号5~8、序列号9~12、序列号13~16或序列号17~20分别表示的FR1、FR2、FR3及FR4至少60%、70%、80%、90%或95%的同一性的氨基酸序列的组合。
另外,只要具有规定的抗原特异性,对于分别由上述序列号1~4、序列号5~8、序列号9~12、序列号13~16或序列号17~20表示的FR1、FR2、FR3及FR4,骨架区可以是具有1~数个、优选1~20个、1~10个、1~5个、1~4个、1~3个、1或2个缺失、置换、附加或插入的氨基酸序列的组合。
CDR区根据靶蛋白确定。通过将针对靶蛋白的公知的VHH抗体或针对后述筛选中检索到的靶抗原的VHH抗体的CDR区域移植到上述FR区,可以设计新的VHH抗体。
另外,在本发明中,“靶蛋白”是指低分子抗体(VHH抗体等)特异性结合的蛋白,与其大小无关。本说明书中,“靶蛋白”这一表达可以与靶肽、靶多肽、靶抗原这些术语互换使用。
“D-氨基酸”
本发明的低分子抗体由D-氨基酸构成。但是,由于甘氨酸没有不对称碳,不存在D型和L型的立体异构(非手性甘氨酸),因此更准确地说,本发明的低分子抗体由D-氨基酸和非手性甘氨酸构成。本说明书中,也可简略并描述为“由D-氨基酸构成”,但其包括“由D-氨基酸和非手性甘氨酸构成”。
构成生物体的氨基酸为L型,D-氨基酸是细菌的肽聚糖等极其有限的物质。由D-氨基酸构成的蛋白质(D型、D-蛋白质)对于具有由L-氨基酸构成的相同序列的蛋白质(L型、L-蛋白质)具有镜像结构(前述Mandal,K.等人、Uppalapati,M.等人))。据报道,生物体中存在的酶区分L型和D型,立体特异性地作用于L-蛋白质,因此D-蛋白质具有酶促降解抗性,是非免疫原性的。
发明人等确认了由D-氨基酸构成的低分子抗体与由L-氨基酸构成的低分子抗体呈镜像的关系。另外,为了将能够作用于生物体中存在的靶蛋白的D-VHH抗体(图1)应用于制药研究,进行了各种研究。在建立了D-VHH抗体的化学合成工艺的同时,确认了D-VHH抗体是非免疫原性的。
2.本发明的低分子抗体的合成
本发明的低分子抗体可以使用D-氨基酸和非手性甘氨酸进行化学合成。由于可通过固相法(SPPS)合成的氨基酸的长度有限,因此将低分子抗体分割成若干片段,将合成的各片段依次连结进行合成。各片段的连结可以通过公知的肽连结方法来实施,但期望利用自然化学连接(Native Chemical Ligation:NCL)法来进行。
“NCL(自然化学连接)法”
NCL法是将在C末端具有α硫酯的肽和在N末端具有未保护半胱氨酸的肽在温和条件下(pH 7、20℃~37℃)混合就可以进行反应,使2个未保护的肽段结合的方法(Dawson,P.E.;Muir,T.W.;Clark-Lewis,I.;Kent,S.B.H.Science,1994,266,776.)。该方法利用肽中本身就具有的官能团,因此不需要特别的活化试剂,即使是侧链无保护的肽,也能高效反应得到良好的产率。
NCL法中,需要靶氨基酸序列中含有半胱氨酸残基的连接位点以适当的间隔存在,但也开发了在半胱氨酸残基极少的蛋白质或在适当的位置不存半胱氨酸残基的情况下,使用丙氨酸残基代替半胱氨酸残基的方法(Johnson,E.C.B.;Kent,S.B.H.J.Am.Chem.Soc.2006,128,6640)。根据NCL法,可以化学合成高达约200个残基的肽链。
本发明的低分子抗体的制造方法包括以下工序。
1)将低分子抗体的氨基酸序列分割为以半胱氨酸残基或丙氨酸残基为N末端的多个片段;
2)通过固相法化学合成各片段;以及
3)通过NCL法将合成的各片段依次连结,合成由D-氨基酸和非手性甘氨酸构成的低分子抗体。
由于通过固相法的化学合成的限制,各片段一般为50个氨基酸及以下、优选为40个氨基酸及以下、更优选为30个氨基酸及以下。使用的固相法没有特别限定,可以利用Boc法、Fmoc法等公知的方法,但优选Fmoc法。为了通过NCL法进行连结,期望各片段的N末端为半胱氨酸残基,但在适当的位置不存在半胱氨酸残基的情况下,如上所述,使用丙氨酸残基。
合成的各片段,在溶解性低的情况下,期望使可溶性标签结合在末端。作为可溶性标签,例如,优选His标签。
在VHH抗体情况下,2个半胱氨酸残基存在于骨架区FR1和FR3中(图2)。但是,用这2个半胱氨酸残基分割的3个片段,中央片段的长度超过70个氨基酸,很难一次性进行化学合成。因此,除了上述2个半胱氨酸残基之外,通过利用存在于它们之间的适当位置的丙氨酸残基或CDR区中的丙氨酸或半胱氨酸残基,将整体分割成50个氨基酸及以下的4个片段进行合成,通过NCL法进行连结。在后述的实施例中,用Cys22、Cys96、Ala54(数值为序列表中的位置)分割为4个片段,这只是一个例子,特别是第2分割点根据VHH抗体的结构(序列)适当设定。
各片段的连结顺序根据合成的抗体或各片段的结构适当确定。在后述的实施例中,预先连结用Ala残基进行了2分割的片段,但也可以连结两端的片段,最后连结Ala部分。在片段的缩合反应中,优选进行产生的硫酯中间体的分离。
合成的低分子抗体在利用尿素等改性后,通过透析或稀释,自然地折叠,形成正确的立体结构。
3.本发明的低分子抗体的筛选
根据L-蛋白质和D-蛋白质的底物特异性的镜像关系,可以通过虚拟镜像库进行制药筛选。具体而言,(1)首先化学合成靶蛋白分子的D型(D-蛋白质);(2)使用合成D-蛋白质筛选手性天然产物;(3)合成命中化合物的镜像(L-蛋白质);以及(4)通过评价L-蛋白质的生物活性可以选择制药候选。
“基于镜像型靶蛋白的筛选”
由D-氨基酸构成的低分子抗体与由L-氨基酸构成的低分子抗体呈完全镜像的关系,靶抗原也呈完全镜像的关系。因此,通过应用通过上述虚拟镜像库进行的筛选,针对天然靶蛋白的、由D-氨基酸构成的低分子抗体的筛选成为可能。
本发明还提供了一种利用上述策略,针对靶抗原蛋白的、由D-氨基酸构成的低分子抗体的制造方法。具体地,所述方法包括以下工序。
1)提供针对靶蛋白的、由D-氨基酸和非手性甘氨酸构成的镜像型靶蛋白;
2)使用所述镜像型靶蛋白筛选抗体库,取得对所述镜像型靶蛋白具有亲和力的抗体;以及
3)基于得到的抗体的氨基酸序列,合成由D-氨基酸和非手性甘氨酸构成的低分子抗体。
作为抗体库,可以使用噬菌体展示库等本领域公知的抗体库,可以是免疫库、天然库、合成库中的任一种。对于scFv或VHH抗体等低分子抗体,已经构建了噬菌体展示库,它们可以被适当利用,也可以按照常规方法制备。
使用噬菌体展示库时,与使用普通靶抗原时同样,可以使用D-靶蛋白实施噬菌体的选择(生物淘选)。即,使库与固定的靶蛋白反应,洗涤除去不结合的噬菌体后,洗脱结合的噬菌体,重复使其在大肠杆菌内增殖的操作,浓缩靶蛋白特异性的噬菌体。固定可以利用通过吸附的固定,或使用生物素-链霉亲和素的固定。
选出的低分子抗体可以根据需要实施亲和力成熟以提高亲和力。亲和力成熟可以是in vivo或in vitro,但期望是in vitro。在in vitro的情况下,也可以通过chainshuffling、利用易错PCR的随机突变导入法、CDR walking等公知的方法将突变导入抗体基因来制备二次库,从中筛选亲和力高的抗体。
确定所选择的天然型低分子抗体的序列,基于该序列化学合成由D-氨基酸构成的低分子抗体。化学合成可以根据前项“3.本发明的低分子抗体的合成”的记载,通过NCL法实施。
如前所述,CDR区根据靶蛋白确定。因此,即使在筛选对象的抗体不是合成的低分子抗体的情况下,也可以通过将该抗体的CDR区移植到合成的低分子抗体的FR区来设计低分子抗体。
4.本发明的低分子抗体的多聚体化
本发明的低分子抗体可以通过用简单的肽键或接头连结、进行多聚体化,制成多价抗体或多特异性抗体。这种多价抗体或多特异性抗体也在本发明的范围内。
例如,可以用肽键连结scFv以制备(scFv)2。另外,可以将scFV二聚体化而形成双抗体、三聚体化而形成三抗体、四聚体化而形成四抗体。进一步地,可以制备更复杂的串联双抗体或柔性抗体(flexibody)。同样地,VHH等单一结构域抗体也可以用简单的肽键或接头串联连结、进行多聚体化。这样构成的多价抗体通过使构成的单体的可变结构域不同,可以作为对不同的靶蛋白具有结合性的多特异性抗体。
作为接头,可以利用由氨基酸构成的肽接头,例如可以使用由Gly和Ser组成的接头。肽接头的长度为1~50个氨基酸、优选1~30个氨基酸、更优选1~10个氨基酸。
或者,也可以利用化学交联剂(合成化学接头)进行连结。作为可用的化学交联剂可以举出NHS、DSS、BS3、DSP、DTSSP、EGS、磺基-EGS、DST、磺基-DST、BSOCOES、磺基-BSOCOES等。
5.本发明的低分子抗体的修饰
本发明的低分子抗体可以根据使用目的适当修饰。作为修饰可以举出通过荧光标记、磷光标记、化学发光标记、生物发光标记、放射性同位素、金属、金属螯合物、金属阳离子、发色团、酶、脂质、亲水性高分子和糖链进行修饰。
修饰还包括用于与药物偶联的接头或间隔物导入的修饰,以及用于稳定化的化学修饰。因此,只要在不对低分子抗体的特异性或立体结构造成负面影响的范围内,也可以将1个或几个氨基酸置换为D-氨基酸以外的非天然型氨基酸或L-氨基酸。
另外,本发明的低分子抗体可以根据使用目的固定在适当的表面上。作为固相化的载体,可以举出胶体颗粒、磁性颗粒、微板等。
6.低分子抗体的利用
6.1治疗性利用
认为由D-氨基酸构成的蛋白质难以受到生物体内肽酶的底物识别,因此难以引起作为抗原呈递细胞(APC)中的片段化的结果而产生的抗原呈递。因此,与天然型(L-)的低分子抗体相比较,镜像型(D-)的低分子抗体难以受到抗原呈递引起的T细胞的活性化,有望抑制被活性化的T细胞引起的B细胞的分化,降低抗药物抗体(ADA)的产生风险。这样一来,具有与天然型的抗体分子同样的特异性、同时免疫原性进一步降低的本发明的低分子抗体,可以单独或与其他药剂连结作为药物使用。
“药物组合物”
在本发明的低分子抗体与靶蛋白结合并发挥规定的药理作用的情况下,可以将该低分子抗体与药理学上容许的载体或添加物一起制剂化,作为药物组合物使用。
作为药理学上容许的载体或添加物,例如可以举出表面活性剂、赋形剂、着色剂、调味剂、保存剂、抗氧化剂、稳定剂、缓冲剂、悬浮剂、等渗剂、粘合剂、崩解剂、润滑剂、流动性促进剂、矫味剂等,但不限于此。
具体地,作为水性载体包括水、乙醇、多元醇(甘油、丙二醇、聚乙二醇等)、橄榄油等植物油、以及油酸乙酯等有机酯。
作为非水性载体,可以举出轻质无水硅酸、乳糖、结晶纤维素、甘露醇、淀粉、羧甲基纤维素钙、羧甲基纤维素钠、羟丙基纤维素、羟基丙基甲基纤维素、聚乙烯缩醛二乙基氨基乙酸酯、聚乙烯吡咯烷酮、明胶、中链脂肪酸甘油三酯、聚氧乙烯氢化蓖麻油60、蔗糖、羧甲基纤维素、玉米淀粉、无机盐类等。
“抗体药物偶联物(ADC)”
通过抗体的特异性靶向识别功能,经由接头将本发明的低分子抗体与适当的药物偶联,可用作抗体药物偶联物(Antibody Drug Conjugate:ADC)。
例如,如果制备与肿瘤细胞特异性抗原结合的低分子抗体并与抗癌剂偶联,并设计成使接头在肿瘤细胞内裂解,则可以使抗癌剂在肿瘤细胞内特异性地发挥作用(内在化ADC)。另外,通过使用在肿瘤细胞外的肿瘤微环境中裂解的接头,也可以使抗癌剂在肿瘤细胞附近作用(外在性ADC)。在某些情况下,也可以使用不裂解的接头。可裂解接头例如基于二硫化物、腙、肽设计,不可裂解接头可基于硫醚(不可切割)等设计。接头可以根据目的适当设计,这样的技术在本领域也是公知的。
含有本发明的低分子抗体的ADC可以与药理学上容许的载体或添加物一起制剂化,这样的载体或添加物如上所述。
含有本发明的低分子抗体的药物组合物或ADC的给药途径没有特别限定,优选胃肠外给药,具体地,可以举出注射给药、经鼻给药、经肺给药、经皮给药等。作为注射给药,可以例示静脉内注射、肌肉内注射、腹腔内注射、皮下注射。给药方法可以根据患者的年龄、症状适当选择。
含有本发明的低分子抗体的药物组合物或ADC的给药量根据作为目的的治疗效果、给药方法、治疗期限、患者的年龄及体重等进行调整,以提供最佳的应答(例如,治疗应答),但通常成人每日活性成分为10μg/kg~10mg/kg。典型的治疗方法包括:例如,每周给药一次,每隔2周给药一次,每隔3周给药一次,每隔4周给药一次,每月给药一次,每隔3个月给药一次或每隔3~6个月给药一次。
6.2诊断性利用
本发明的低分子抗体在生物体内的稳定性高,免疫原性也低,因此通过用可检测的分子进行修饰,可以用于诊断和成像。
作为可检测的分子,例如可以举出放射性核素:铯99m(99mTc)、碘123(123I)、碘125(125I)、铟111(111In)、氟18(18F)、镓67(67Ga)、镓68(68Ga)、铜64(64Cu)等;荧光分子:荧光素、Alexa、花青等;化学发光分子:鲁米诺等;生物发光分子:荧光素酶、碱性磷酸酶等;等等。
用可检测的分子进行修饰的低分子抗体可以根据修饰分子通过放射免疫分析(RIA)、辅酶免疫分析(EIA)、荧光免疫分析(FIA)、发光免疫分析(LIA)、免疫沉淀法(IP)、免疫比浊法(TIA)、蛋白质免疫印迹(WB)、免疫组织化学(IHC)法、免疫扩散法(SRID)检测。
7.低分子抗体的制造用试剂盒
在VHH抗体中,3个CDR区中CDR3最具多样性,对于与靶的相互作用很重要,其他2个CDR被认为对相互作用的影响很小,可以作为固定序列。因此,可以预先准备到FR3区的半胱氨酸(Cys96)之前的片段(后述的实施例的D-9)或构成所述片段的部分片段,作为VHH抗体试剂盒的制造用试剂盒提供。其余片段可根据靶向设计合成,并通过上述NCL法与试剂盒的片段连结以制造靶向特异性VHH抗体。
例如,本发明的试剂盒包括分别包含下述(a)或(b)的氨基酸序列的、由D-氨基酸和非手性甘氨酸构成的一个或多个多肽。
(a)由序列号55~57、61及63中任一个表示的氨基酸序列;
(b)具有与由序列号55~57、61及63中任一个表示的氨基酸序列(优选在与所述序列的骨架区对应的部分中)至少60%、70%、80%、90%、95%、98%或99%的同一性的氨基酸序列。
作为(b)的氨基酸序列,例如可以举出将相当于骨架区的部分的序列置换为其他公知的VHH抗体的骨架区(例如,参照序列号5~20或序列号21~54(图8))的对应的序列的序列。
优选地,本发明的试剂盒包括:
(1)含有由作为到FR3区的半胱氨酸(Cys96)之前的片段的序列号63(实施例的D-9)表示的氨基酸序列的、由D-氨基酸和非手性甘氨酸构成的多肽,或者
(2)含有由作为不含CDR区的FR1区的一部分的序列号55(实施例的D-1)表示的氨基酸序列的、由D-氨基酸和非手性甘氨酸构成的多肽
或具有具有与上述氨基酸序列、优选在与该骨架区对应的部分中至少60%、70%、80%、90%、95%、98%或99%的同一性的氨基酸序列的多肽。
各多肽可以通过保护基、组氨酸标签或荧光标记等标记适当修饰,也可以固定在珠子或支持体等固相上。关于修饰和固定,参照5的记载。
除了作为必要的构成要素的上述多肽之外,试剂盒还可以包括合成所需的试剂或缓冲液、D-氨基酸或非手性甘氨酸、使用说明书等。
实施例
以下,通过实施例对本发明进行具体地说明,但本发明不限于这些实施例。
实施例1
在本实施例中,通过图3所示的工艺合成了GFP结合VHH抗体(L-GFP-VHH抗体)及其镜像体(D-GFP-VHH抗体)。
(1)HPLC及MS
分析用高效液相色谱(HPLC)是将COSMOSIL 5C18-AR300色谱柱(4.6×250mm、Nacalai Tesque)在流速1mL/min(25℃)、含有0.1%(v/v)TFA的CH3CN的线性梯度下使用。L-10和D-10的分析中,将COSMOSIL 5C4-AR300色谱柱(4.6×150mm、Nacalai Tesque)在流速1mL/min(40℃)、含有0.1%(v/v)TFA的CH3CN的线性梯度下使用。产物在220nm处检测吸光度。
在HPLC纯化中,将COSMOSIL 5C18-AR300色谱柱(20×250mm、Nacalai Tesque)或COSMOSIL 5C4-AR300色谱柱(20×150mm、Nacalai Tesque)在流速8mL/min(室温)、含有0.1%(v/v)TFA的CH3CN的线性梯度下使用。L-10和D-10中,将COSMOSIL5C4-AR300色谱柱(20×150mm、Nacalai Tesque)在流速12mL/min(40℃)下、含有0.1%(v/v)TFA的CH3CN的线性梯度下使用。
(2)Fmoc-SPPS
通过Fmoc法的肽固相合成(Fmoc-SPPS)使用自动肽合成装置(PSSM-8、岛津)进行。侧链保护氨基酸使用以下物质:Arg(Pbf)、Asn(Trt)、Asp(OtBu)、Cys(Trt)、Gln(Trt)、Glu(OtBu)、His(Trt)、Lys(Boc)、Ser(tBu)、Thr(tBu)、Tyr(tBu)。Fmoc保护氨基酸(5当量)使用HBTU(5当量)、HOBt·H2O(5当量)和(i-Pr)2NEt(10当量),在DMF中进行45分钟2次偶联。在20%哌啶/DMF中4分钟2次脱保护Fmoc保护基。
(3)L-1的合成
通过上述Fmoc-SPPS的标准程序,从Fmoc-NH-SAL树脂(45mg、0.02mmol)开始构建肽序列。构建肽链后,在DMF中Boc2O(22mg、0.10mmol)和(i-Pr)2NEt(35μL、0.20mmol)中反应2小时,进行N末端胺保护。洗涤后,将树脂在DCM(2mL)中50mM 4-硝基苯基氯甲酸酯中处理1小时后,在DMF(2mL)中的0.5M(i-Pr)2NEt中处理15分钟。在TFA/H2O/m-甲酚/硫代茴香醚/EDT(80:5:5:5:5)中进行2小时的脱保护和与树脂的切割。通过过滤除去树脂后,使粗品沉淀,用冷Et2O洗涤。将得到的沉淀物溶解在最少量的MESNa缓冲液(500mM MESNa、6M胍、200mM磷酸缓冲液:pH 6.0)中,孵育1小时。通过HPLC纯化粗品,得到所需的肽L-1(9.1mg、产率21%:序列号55)。
MS(ESI):C92H159N27O32S2的计算值:2219.56;测定值(m/z):[M+2H]2+=1110.60。
(4)L-2的合成
通过上述Fmoc-SPPS的标准程序,从Fmoc-NH-SAL树脂(45mg、0.02mmol)开始构建保护肽。肽L-2的N末端半胱氨酸的偶联包括将Boc-Cys(Acm)-OH与HBTU(38mg、0.10mmol)、HOBt·H2O(15mg、0.10mmol)和(i-Pr)2NEt(35μL、0.20mmol)一起使用。洗涤后,将树脂在DCM(2mL)中50mM 4-硝基苯基氯甲酸酯中处理1小时后,在DMF(2mL)中的0.5M(i-Pr)2NEt中处理15分钟。在TFA/H2O/m-甲酚/硫代茴香醚/EDT(80:5:5:5:5)中处理2小时,进行脱保护和与树脂的切割。通过过滤除去树脂后,使粗品沉淀,用冷Et2O洗涤。将得到的沉淀物溶解在最少量的MESNa缓冲液(500mM MESNa、6M胍、200mM磷酸缓冲液:pH 6.0)中,孵育1小时。通过HPLC纯化粗品,得到所需的肽L-2(9.3mg、产率12%:序列号56)。
MS(ESI):C166H250N50O48S5的计算值:3874.43;测定值(m/z):[M+4H]4+=969.56、[M+3H]3+=1292.52、[M+2H]2+=1937.87。
(5)L-3的合成
通过上述Fmoc-SPPS的标准程序,从Fmoc-NH-SALPEG树脂(87mg、0.02mmol)开始构建保护肽。在TFA/H2O/m-甲酚/硫代茴香醚/EDT(80:5:5:5:5)中进行2小时的脱保护和与树脂的切割。通过过滤除去树脂后,使粗品沉淀,用冷Et2O洗涤。将得到的沉淀物溶解在最少量的含有0.1%TFA的50%CH3CN溶液中。通过HPLC纯化粗品,得到所需的肽L-3(11mg、10%产率:序列号57)。
MS(ESI):C220H340N66O73S2的计算值:5141.65;测定值(m/z):[M+6H]6+=858.17、[M+5H]5+=1029.63、[M+4H]4+=1286.57、[M+3H]3+=1714.78。
(6)L-4的合成
通过上述Fmoc-SPPS的标准程序,从Fmoc-NH-SAL树脂(93mg、0.04mmol)开始构建保护肽。在TFA/H2O/m-甲酚/硫代茴香醚/EDT(80:5:5:5:5)中进行2小时的脱保护和树脂的切割。通过过滤除去树脂后,使粗品沉淀,用冷Et2O洗涤。将得到的沉淀物溶解在最少量的含有0.1%TFA的50%CH3CN溶液中。通过HPLC纯化粗品,得到所需的肽L-4(3.6mg、2.8%产率:序列号58)。
MS(ESI):C137H194N46O38S的计算值:3125.40;测定值(m/z):[M+4H]4+=782.59、[M+3H]3+=1043.02、[M+2H]2+=1564.07。
(7)L-5的合成
使硫酯L-2(7.7mg、2.0μmol)和肽L-3(11mg、2.1μmol)在连接溶液(400mM MPAA、100mM TCEP、6M胍、200mM磷酸缓冲液:pH 7.0;1.0mL)中在37℃下反应2小时。通过分析用HPLC监测反应。通过HPLC纯化粗品,得到所需的肽L-5(12mg、70%产率:序列号59)。
MS(ESI):C384H584N116O118S5的计算值:8873.89;测定值(m/z):[M+11H]11+=808.22、[M+10H]10+=888.67、[M+9H]9+=987.29、[M+8H]8+=1110.53、[M+7H]7+=1268.91、[M+6H]6+=1480.28、[M+5H]5+=1775.91。
(8)L-6的合成
使肽L-5(13mg、1.5μmol)在脱硫用溶液(20mM VA-044、100mM MESNa、250mM TCEP、6M胍、100mM磷酸缓冲液:pH 6.5;3.0mL)中在37℃下处理2小时。通过分析用HPLC监测反应。通过HPLC纯化粗品,得到所需的肽L-6(11mg、85%产率:序列号60)。
MS(ESI):C384H584N116O118S4的计算值:8841.83;测定值(m/z):[M+8H]8+=1106.64、[M+7H]7+=1264.47、[M+6H]6+=1474.84、[M+5H]5+=1769.45。
(9)L-7的合成
使肽L-6(6.9mg、0.78μmol)和PdCl2(1.4mg、7.8μmol)在含有6M胍(390μL)的100mM磷酸缓冲液中在37℃下反应30分钟。然后加入少量的DTT。通过分析用HPLC监测反应。通过HPLC纯化粗品,得到所需的肽L-7(3.9mg、产率58%:序列号61)。
MS(ESI):C381H579N115O117S4的计算值:8770.75;测定值(m/z):[M+11H]11+=798.56、[M+10H]10+=878.51、[M+9H]9+=975.89、[M+8H]8+=1097.46、[M+7H]7+=1254.52、[M+6H]6+=1462.83、[M+5H]5+=1755.07。
(10)L-8的合成
使硫酯L-1(1.8mg、0.80μmol)和肽L-7(2.3mg、0.27μmol)在连接缓冲液(400mMMPAA、100mM TCEP、6M胍、200mM磷酸缓冲液:pH 7.0;0.14mL)中在37℃下反应1小时。通过分析用HPLC监测反应。通过HPLC纯化粗品,得到所需的肽L-8(1.8mg、产率62%:序列号62)。
MS(ESI):C471H732N142O146S4的计算值:10848.13;测定值(m/z):[M+13H]13+=835.72、[M+12H]12+=905.33、[M+11H]11+=987.56、[M+10H]10+=1086.31、[M+9H]9+=1206.72、[M+8H]8+=1357.45、[M+7H]7+=1551.12、[M+6H]6+=1809.18。
(11)L-9的合成
使肽L-8(1.1mg、0.10μmol)在活化溶液(133mM NaNO2、6M胍、200mM磷酸缓冲液:pH3.0;37.5μL)中在-20℃下活化30分钟。添加连接溶液(400mM MPAA、100mM TCEP、6M胍、200mM磷酸缓冲液:pH 7.0;12.5μL)并进行硫酯化。通过分析用HPLC监测反应。通过HPLC纯化粗品,得到所需的肽L-9(0.66mg、产率61%:序列号63)。
MS(ESI):C466H719N135O146S5的计算值:10708.98;测定值(m/z):[M+12H]12+=893.56、[M+11H]11+=974.98、[M+10H]10+=1072.25、[M+9H]9+=1191.34、[M+8H]8+=1339.96、[M+7H]7+=1530.98、[M+6H]6+=1785.87。
(12)L-10(合成L-GFP-VHH抗体)的合成
使硫酯L-9(7.6mg、0.71μmol)和肽L-4(5.2mg、1.7μmol)在连接溶液(400mM MPAA、100mM TCEP、6M胍、200mM磷酸缓冲液:pH 7.0;1.1mL)中室温反应10小时。通过分析用HPLC监测反应。通过HPLC纯化粗品,得到所需的肽L-10(6.0mg、产率62%:序列号64)。
MS(ESI):C595H905N181O182S5的计算值:13666.17;测定值(m/z):[M+14H]14+=977.81、[M+13H]13+=1052.81、[M+12H]12+=1140.62、[M+10H]10+=1368.11、[M+9H]9+=1519.85、[M+8H]8+=1709.45、[M+7H]7+=1954.09。
(13)D-1的合成
通过与上述L-1的合成相同的工艺,得到D-1(60.9mg、产率20%:序列号55(氨基酸除了非手性甘氨酸以外全部为D型,以下同样)。
MS(ESI):C92H159N27O32S2的计算值:2219.56;测定值(m/z):[M+2H]2+=1110.97。
(14)D-2的合成
通过与上述L-2的合成相同的工艺,得到D-2(68.6mg、产率13%:序列号56)。
MS(ESI):C166H250N50O48S5的计算值:3874.43;测定值(m/z):[M+4H]4+=969.87、[M+3H]3+=1292.76、[M+2H]2+=1938.16。
(15)D-3的合成
通过与上述L-3的合成相同的工艺,得到D-3(7.2mg、产率7.0%:序列号57)
MS(ESI):C220H340N66O73S2的计算值:5141.65;测定值(m/z):[M+6H]6+=858.33、[M+5H]5+=1029.56、[M+4H]4+=1286.77、[M+3H]3+=1715.34。
(16)D-4的合成
通过与上述L-4的合成相同的工艺,得到D-4(10.2mg、产率2.3%:序列号58)。
MS(ESI):C137H194N46O38S的计算值:3125.40;测定值(m/z):[M+4H]4+=782.66、[M+3H]3+=1043.10、[M+2H]2+=1563.31。
(17)D-5的合成
通过与上述L-5的合成相同的工艺,得到D-5(13.8mg、产率65%:序列号59)。
MS(ESI):C384H584N116O118S5的计算值:8873.89;测定值(m/z):[M+11H]11+=808.08、[M+10H]10+=888.83、[M+9H]9+=987.60、[M+8H]8+=1110.83、[M+7H]7+=1269.51、[M+6H]6+=1480.21、[M+5H]5+=1776.25。
(18)D-6的合成
通过与上述L-6的合成相同的工艺,得到D-6(3.7mg、产率47%:序列号60)。
MS(ESI):C384H584N116O118S4的计算值:8841.83;测定值(m/z):[M+8H]8+=1106.65、[M+7H]7+=1264.88、[M+6H]6+=1474.73、[M+5H]5+=1769.45。
(19)D-7的合成
通过与上述L-7的合成相同的工艺,得到D-7(3.1mg、产率63%:序列号61)。
MS(ESI):C381H579N115O117S4的计算值:8770.75;测定值(m/z):[M+11H]11+=798.58、[M+10H]10+=878.33、[M+9H]9+=975.88、[M+8H]8+=1097.92、[M+7H]7+=1254.38、[M+6H]6+=1463.04、[M+5H]5+=1755.13。
(20)D-8的合成
通过与上述L-8的合成相同的工艺,得到D-8(3.6mg、产率75%:序列号62)。
MS(ESI):C471H732N142O146S4的计算值:10848.13;测定值(m/z):[M+13H]13+=835.71、[M+12H]12+=905.52、[M+11H]11+=987.80、[M+10H]10+=1086.24、[M+9H]9+=1207.04、[M+8H]8+=1357.72、[M+7H]7+=1551.36、[M+6H]6+=1809.89。
(21)D-9的合成
通过与上述L-9的合成相同的工艺,得到D-9(7.1mg、产率66%:序列号63)。
MS(ESI):C466H719N135O146S5的计算值:10708.98;测定值(m/z):[M+11H]11+=975.07、[M+10H]10+=1072.56、[M+9H]9+=1191.18、[M+8H]8+=1340.21、[M+7H]7+=1531.30、[M+6H]6+=1786.13。
(22)D-10(合成D-GFP-VHH抗体)的合成
通过与上述L-10的合成相同的工艺,得到D-10(3.8mg、产率42%:序列号64)。
MS(ESI):C595H905N181O182S5的计算值:13666.17;测定值(m/z):[M+14H]14+=977.48、[M+13H]13+=1052.68、[M+12H]12+=1140.47、[M+9H]9+=1519.70、[M+8H]8+=1710.32、[M+7H]7+=1953.53。
(23)重组蛋白的制备
重组L-GFP-VHH抗体的质粒通过将C末端6×His克隆到pGEX6P1-GFP-Nanobody(Addgene#61838)来制备。将所有质粒转化为BL21(DE3)细胞并在LB培养基中培养。大肠杆菌在含有氨苄西林(最终浓度50μg/mL)的LB培养基中在37℃下培养过夜,添加100mM IPTG并在20℃下表达过夜。大肠杆菌经离心分离(6000rpm、20分钟、4℃)后再悬浮于结合缓冲液(PBS中5mM DTT及蛋白酶抑制剂[Nacalai Tesque])中。超声处理及添加TritionX-100后,将细胞溶解物在4℃下、以12000rpm离心分离30分钟。将上清液与谷胱甘肽-琼脂糖4B(GEHealthcare)一起在4℃下孵育过夜。为了纯化重组L-GFP-VHH抗体,通过在切割缓冲液(50mM Tris-HCl:pH 7.5、150mM NaCl、1mM EDTA、1mM DTT、0.01%TritonX-100)中加入PreScission蛋白酶(GEHelthcare)并在4℃下孵育过夜来除去GST标签。为了纯化GST-EGFP和GST-mCherry,将带GST标签的蛋白通过洗脱缓冲液(含有10mM谷胱甘肽的PBS:pH 8.5)在4℃下洗脱30分钟。GST-mCherry蛋白通过使用PBS的凝胶过滤层析(Superdex75 Increase、GE Healthcare)进一步纯化。蛋白的纯度全部用SDS-PAGE确认。
(24)合成L-GFP-VHH抗体及合成D-GFP-VHH抗体的折叠
使肽L-10或D-10溶解在含有6M胍和40mM DTT的PBS(pH 7.4)中至1.0mg/mL,室温孵育2小时。将该溶液用PBS(pH 7.4)稀释100倍,室温放置过夜。使用MWCO3000离心滤膜(Millipore、Amicon-Ultra3kDa)浓缩溶液。
(25)重组L-GFP-VHH抗体、合成L-GFP-VHH抗体及合成D-GFP-VHH抗体的CD光谱
将重组L-GFP-VHH抗体、合成L-GFP-VHH抗体及合成D-GFP-VHH抗体用PBS(pH 7.4)稀释,将浓度分别调整为10μM。各蛋白质的CD光谱使用圆二色光谱仪(JACSO J-720)在20℃下记录。各蛋白质的热稳定性通过在203nm的波长下测定37℃和95℃的变化来推测。
合成L-GFP-VHH抗体的CD光谱与重组L-GFP-VHH抗体的CD光谱一致,暗示存在β-折叠结构(图4A)。合成D-GFP-VHH抗体的CD光谱与重组体L-GFP-VHH抗体的CD光谱相反。合成L-及D-GFP-VHH抗体的热稳定性与重组L-GFP-VHH抗体的热稳定性相同。暗示D-VHH抗体支架在生理学条件下是稳定的(图4B)。这些结果表明合成L-及D-GFP-VHH抗体两者均被正确折叠。
(26)酶联免疫吸附测定法(ELISA)
ELISA使用含有0.025% Tween20的PBS(pH 7.4)实施所有洗涤和稀释过程。使用50mM碳酸钠缓冲液(pH 9.4)中的GST-EGFP或GST-mCherry在4℃下对96孔微孔板(Greiner、high binding)进行包衣过夜(50μL/孔;30nM)。包衣处理后,将孔洗涤3次,用含有3%BSA的PBS(150μL/孔)封闭2小时。洗涤3次后,添加重组或合成的L-GFP-VHH抗体(50μL/孔,0、0.03、0.1、0.3、1、3、10、30nM),孵育1小时。洗涤3次后,加入抗组氨酸单克隆抗体(28-75)(和光)(50μL/孔),在4℃下孵育1小时。洗涤3次后,添加抗小鼠IgG(H+L)HRP偶联物(Promega)(50μL/孔),在4℃下孵育1小时。洗涤5次后,添加TMB溶液并孵育15分钟,使用1MH2SO4(50μL/孔)停止反应。使用iMark Microplate Absorbance Reader(Bio-Rad)在450nm处测定吸光度。
重组及合成L-GFP-VHH抗体两者都与GST-EGFP结合但不与GST-mCherry结合,这暗示合成L-GFP-VHH抗体具有与重组L-GFP-VHH抗体同样的生物学活性(图5)。
(27)水晶振子微平衡(QCM)分析
QCM分析使用Single-Q(SCINICS、日本)实施。传感器芯片的金表面用水虎鱼(piranha)溶液洗涤,加入溶解在10% EtOH中的0.2mM NTA-SAM形成试剂(Dojindo、日本),孵育过夜。洗涤后,加入40mM NiSO4溶液,孵育1小时。用500μL的PBS(pH 7.4)洗涤及平衡后,添加5μL的1.0mg/mL配体溶液(PBS中的重组L-GFP-VHH抗体、合成L-GFP-VHH抗体或合成D-GFP-VHH抗体)。用500μL的PBS(pH 7.4)洗涤及平衡后,反复添加各浓度的样本溶液(最终浓度:0.1nM、0.3nM、1nM、3nM、10nM、30nM、100n300 nM)。解离常数通过3次分析进行评价。
重组及合成L-GFP-VHH抗体与GST-EGFP相互作用,但完全没有观察到与GST-mCherry的结合(图6)。合成D-GFP-VHH抗体也同样固定在Ni-NTA-SAM表面,但没有观察到合成D-GFP-VHH抗体与GST-EGFP之间的相互作用。
从这些结果确认了合成L-GFP-VHH抗体的GFP结合活性。还发现合成D-GFP-VHH抗体具有L-GFP-VHH抗体的完全镜像结构,具有不同的相互作用面。
(28)免疫化及免疫原性分析
在第0天以50μg/injection向每组6只BALB/c小鼠(雌性、6周龄)腹腔内注射用弗氏完全佐剂(Wako)乳化的合成L-或D-GFP-VHH抗体抗原,在第14天、第21天以50μg/注射注入用弗氏不完全佐剂(Wako)乳化的合成L-或D-GFP-VHH抗体抗原。在第0、14、21及28天收集免疫血清。在ELISA所有洗涤和稀释过程中均使用含有0.025% Tween20的PBS(pH 7.4)实施。使用50mM碳酸钠缓冲液(pH 9.4)中的L-GFP-VHH抗体或D-GFP-VHH抗体(50μL/孔;1.0ng/mL、10ng/mL、100ng/mL或1000ng/mL)中的任一种在4℃下对96孔微孔板(Greiner、high binding)进行包衣过夜。包衣处理后,将孔洗涤3次,用含有3%BSA的PBS(150μL/孔)封闭2小时。洗涤3次后,加入各小鼠免疫化血清的1:1000稀释(50μL/孔、0、14、21及28天),孵育1小时。洗涤3次后,加入1:5000稀释的Anti-Mouse IgG(H+L)HRP Conjugate(Promega)(50μL/孔),孵育30分钟。洗涤4次后,添加TMB溶液并孵育15分钟,使用1M H2SO4(50μL/孔)停止反应。使用iMark Microplate Absorbance Reader(Bio-Rad)在450nm处测定吸光度。
在用L-GFP-VHH抗体免疫的所有小鼠的血清(免疫14天后)中观察到针对L-GFP-VHH抗体的抗体的存在(图7)。ADA的浓度通过反复注入依赖于抗原的包衣浓度而增加。对照地,即使在28天后,在用D-GFP-VHH抗体免疫的所有小鼠的血清中也没有观察到针对D-GFP-VHH抗体的抗体的存在。用相反镜像异构体型的抗原免疫小鼠时,没有观察到ADA的出现。这些结果暗示D-GFP-VHH抗体是非免疫原性的,而L-GFP-VHH抗体是免疫原性的。
上述实施例使用抗GFP VHH抗体进行实施,但本领域技术人员可以理解,这些结果也适用于其他靶蛋白。
实施例2
在本实施例中,通过图9所示的工艺合成了VHH抗体PMP12A2h1及其镜像体(D-PMP12A2h1)。PMP12A2h1是构成以卡拉西单抗一般名称已知的药物产品的VHH抗体,与凝血因子之一的von Willebrand因子的A1结构域(vWF A1结构域)特异性结合,抑制血小板聚集。
(1)HPLC及MS
分析用高效液相色谱(HPLC)是将COSMOSIL 5C18-AR300色谱柱(4.6×250mm、Nacalai Tesque)在流速1mL/min(25℃)、含有0.1%(v/v)TFA的CH3CN的线性梯度下使用。产物在220nm处检测吸光度。
在HPLC纯化中,将COSMOSIL 5C18-AR300色谱柱(20×250mm、Nacalai Tesque)或COSMOSIL 5C4-AR300色谱柱(20×150mm、Nacalai Tesque)在流速8mL/min(室温)、含有0.1%(v/v)TFA的CH3CN的线性梯度下使用。
(2)保护肽树脂的构建
通过Fmoc法的肽固相合成(Fmoc-SPPS)使用自动肽合成装置(PSSM-8、岛津或Liberty BLUE、CEM)进行。在使用PSSM-8的合成中,侧链保护氨基酸使用以下物质:Arg(Pbf)、Asn(Trt)、Asp(OtBu)、Cys(Trt)、Gln(Trt)、Glu(OtBu)、His(Trt)、Lys(Boc)、Ser(tBu)、Thr(tBu)、Tyr(tBu)。Fmoc保护氨基酸(5当量)使用Oxyma Pure(5当量)和DIC(10当量),在DMF中进行60分钟2次偶联。在20%哌啶/DMF中4分钟2次脱保护Fmoc保护基。
(3)伴随微波照射的保护肽树脂的构建
在伴随微波照射的肽合成(Liberty BLUE)中,侧链保护氨基酸使用以下物质:Arg(Pbf)、Asn(Trt)、Asp(OtBu)、Cys(Trt)、Gln(Trt)、Glu(OtBu)、His(Trt)、Lys(Boc)、Ser(tBu)、Thr(tBu)、Trp(Boc)、Tyr(tBu)。Fmoc保护氨基酸(5当量)使用Oxyma Pure(5当量)和DIC(10当量),在DMF中125秒90℃微波照射下进行偶联。对于Arg(Pbf),在DMF中125秒90℃微波照射下进行2次偶联。对于His(Trt),在DMF中10分钟50℃微波照射下进行2次偶联。在20%哌啶/DMF中90秒90℃微波照射下脱保护Fmoc保护基。
(4)L-11的合成
通过上述Fmoc-SPPS的标准程序,从Fmoc-NH-SAL树脂(439mg、0.25mmol)开始构建肽序列。按Fmoc-Arg(Pbf)-OH、Fmoc-Dbz-OH、Fmoc-Ser(tBu)-OH的顺序缩合后,通过微波辅助Fmoc-SPPS延伸肽链。构建肽链后,在DMF中Boc2O(273mg、1.25mmol)和(i-Pr)2NEt(435μL、2.5mmol)中反应2小时,进行N末端胺保护。洗涤后,将树脂在DCM(25mL)中50mM 4-硝基苯基氯甲酸酯中处理1小时后,在DMF(25mL)中的0.5M(i-Pr)2NEt中处理15分钟。在TFA/H2O/m-甲酚/硫代茴香醚/EDT(80:5:5:5:5)中进行2小时的脱保护和与树脂的切割。通过过滤除去树脂后,使粗品沉淀,用冷Et2O洗涤。将得到的沉淀物溶解在含有0.1%TFA的50%CH3CN溶液中后,通过HPLC纯化粗品,得到所需的肽L-11(113mg、产率19%:序列号65)。
MS(ESI):C103H169N33O33的计算值:2397.68;测定值(m/z):[M+3H]3+=799.89、[M+2H]2+=1199.22。
(5)L-12的合成
通过上述Fmoc-SPPS的标准程序,从Fmoc-NH-SAL树脂(439mg、0.25mmol)开始构建肽序列。按Fmoc-Arg(Pbf)-OH、Fmoc-MeDbz-OH、Fmoc-Ala-OH的顺序缩合后,通过微波辅助Fmoc-SPPS延伸肽链。N末端半胱氨酸的偶联使用Boc-Cys(Acm)-OH。将构建肽链的树脂在DCM(25mL)中50mM 4-硝基苯基氯甲酸酯中处理1小时后,在DMF(25mL)中的0.5M(i-Pr)2NEt中处理15分钟。在TFA/H2O/m-甲酚/硫代茴香醚/EDT(80:5:5:5:5)中进行2小时的脱保护和与树脂的切割。通过过滤除去树脂后,使粗品沉淀,用冷Et2O洗涤。将得到的沉淀物溶解在含有0.1%TFA的50%CH3CN溶液中后,通过HPLC纯化粗品,得到所需的肽L-12(255mg、产率29%:序列号66)。
MS(ESI):C153H229N49O40S2的计算值:3458.94;测定值(m/z):[M+5H]5+=692.18、[M+4H]4+=865.48、[M+3H]3+=1153.70。
(6)L-13的合成
通过上述Fmoc-SPPS的标准程序,从Fmoc-NH-SAL PEG树脂(91mg、0.02mmol)开始构建肽序列。按Fmoc-Arg(Pbf)-OH、Fmoc-Dbz-OH、Fmoc-Tyr(tBu)-OH的顺序缩合后,通过Fmoc-SPPS延伸肽链。在TFA/H2O/m-甲酚/硫代茴香醚/EDT(80:5:5:5:5)中进行2小时的脱保护和与树脂的切割。通过过滤除去树脂后,使粗品沉淀,用冷Et2O洗涤。将得到的沉淀物溶解在含有0.1%TFA的50%CH3CN溶液中后,通过HPLC纯化粗品,得到所需的肽L-13(3.1mg、产率2.8%:序列号67)。
MS(ESI):C241H374N72O75S3的计算值:5576.25;测定值(m/z):[M+7H]7+=797.20、[M+6H]6+=930.27、[M+5H]5+=1116.27、[M+4H]4+=1394.60、[M+3H]3+=1858.98。
(7)L-14的合成
通过上述Fmoc-SPPS的标准程序,从Fmoc-NH-SAL PEG树脂(91mg、0.02mmol)开始构建保护肽。在TFA/H2O/m-甲酚/硫代茴香醚/EDT(80:5:5:5:5)中进行2小时的脱保护和树脂的切割。通过过滤除去树脂后,使粗品沉淀,用冷Et2O洗涤。将得到的沉淀物溶解在最少量的含有0.1%TFA的50%CH3CN溶液中。通过HPLC纯化粗品,得到所需的肽L-14(6.1mg、8.7%产率:序列号68)。
MS(ESI):C150H236N46O49S的计算值:3499.87;测定值(m/z):[M+3H]3+=1167.23、[M+2H]2+=1750.12。
(8)L-15的合成
使硫酯L-12(17mg、4.8μmol)和肽L-13(30mg、5.3μmol)在连接溶液(400mM MPAA、100mM TCEP、6M胍、200mM磷酸缓冲液:pH 7.0;2.4mL)中在37℃下处理2小时。通过分析用HPLC监测反应。通过HPLC纯化粗品,得到所需的肽L-15(12mg、30%产率:序列号69)。
MS(ESI):C379H582N114O112S5的计算值:8687.81;测定值(m/z):[M+11H]11+=790.42、[M+10H]10+=869.51、[M+9H]9+=966.20、[M+8H]8+=1087.03、[M+7H]7+=1242.50、[M+6H]6+=1448.39、[M+5H]5+=1738.15。
(9)L-16的合成
使肽L-15(77mg、8.9μmol)在脱硫反应溶液(20mM VA-044、100mM MESNa、250mMTCEP、6M胍、200mM磷酸缓冲液:pH 6.5;18mL)中在37℃下处理2小时。通过分析用HPLC监测反应。通过HPLC纯化粗品,得到所需的肽L-16(63mg、82%产率:序列号70)。
MS(ESI):C379H582N114O112S4的计算值:8655.75;测定值(m/z):[M+11H]11+=787.81、[M+10H]10+=866.47、[M+9H]9+=962.45、[M+8H]8+=1083.00、[M+7H]7+=1237.57、[M+6H]6+=1442.96、[M+5H]5+=1731.32。
(10)L-17的合成
使肽L-16(61mg、7.0μmol)和PdCl2(12mg、70μmol)在含有6M胍的200mM磷酸缓冲液(3.5mL)中在37℃下反应30分钟。然后加入少量DTT,离心后回收上清液。通过分析用HPLC监测反应。通过HPLC纯化粗品,得到所需的肽L-17(29mg、产率47%:序列号71)。
MS(ESI):C376H577N113O111S4的计算值:8584.67;测定值(m/z):[M+11H]11+=781.81、[M+10H]10+=859.64、[M+9H]9+=955.10、[M+8H]8+=1074.37、[M+7H]7+=1227.16、[M+6H]6+=1431.14、[M+5H]5+=1717.18。
(11)L-18的合成
使硫酯L-11(22mg、9.3μmol)和肽L-17(27mg、3.1μmol)在连接缓冲液(100mMMPAA、50mM TCEP、6M胍、200mM磷酸缓冲液:pH 7.0;1.5mL)中在37℃下反应1小时。通过分析用HPLC监测反应。通过HPLC纯化粗品,得到所需的肽L-18(21mg、产率64%:序列号72)。
MS(ESI):C465H727N139O141S4的计算值:10649.00;测定值(m/z):[M+13H]13+=819.98、[M+12H]12+=888.21、[M+11H]11+=968.84、[M+10H]10+=1065.79、[M+9H]9+=1184.43、[M+8H]8+=1331.65、[M+7H]7+=1521.62、[M+6H]6+=1774.94。
(12)L-19的合成
使肽L-18(2.2mg、0.21μmol)在活化溶液(133mM NaNO2、6M胍、200mM磷酸缓冲液:pH 3.0;78μL)中在-20℃下活化30分钟。添加连接溶液(400mM MPAA、100mM TCEP、6M胍、200mM磷酸缓冲液:pH 7.0;26μL)并进行硫酯化。通过分析用HPLC监测反应。通过HPLC纯化粗品,得到所需的肽L-19(1.1mg、产率52%:序列号73)。
MS(ESI):C460H714N132O141S5的计算值:10509.86;测定值(m/z):[M+11H]11+=956.78、[M+10H]10+=1051.81、[M+9H]9+=1168.81、[M+8H]8+=1314.32、[M+7H]7+=1502.25、[M+6H]6+=1751.85。
(13)L-20(PMP12A2h1)的合成
使硫酯L-19(1.1mg、0.11μmol)和肽L-14(0.84mg、0.24μmol)在连接溶液(400mMMPAA、100mM TCEP、6M胍、200mM磷酸缓冲液:pH 7.0;55μL)中在37℃下反应2小时。通过分析用HPLC监测反应。通过HPLC纯化粗品,得到所需的肽L-20(1.0mg、产率68%:序列号74)。
MS(ESI):C602H942N178O188S5的计算值:13841.52;测定值(m/z):[M+15H]15+=923.60、[M+14H]14+=989.55、[M+13H]13+=1065.64、[M+12H]12+=1154.64、[M+11H]11+=1259.18、[M+10H]10+=1384.96、[M+9H]9+=1538.53、[M+8H]8+=1731.24。
(14)D-11的合成
通过与上述L-11的合成相同的工艺,得到D-11(115mg、产率19%:序列号65(氨基酸除了非手性甘氨酸以外全部为D型,以下同样)。
MS(ESI):C103H169N33O33的计算值:2397.68;测定值(m/z):[M+3H]3+=799.96、[M+2H]2+=1199.68。
(15)D-12的合成
通过与上述L-12的合成相同的工艺,得到D-12(268mg、产率31%:序列号66)。
MS(ESI):C153H229N49O40S2的计算值:3458.94;测定值(m/z):[M+5H]5+=692.59、[M+4H]4+=865.69、[M+3H]3+=1153.56。
(16)D-13的合成
通过与上述L-13类似的工艺合成。通过上述Fmoc-SPPS的标准程序,从NovaSynTGR树脂(4.0g、1.0mmol)开始构建肽序列。按Fmoc-D-Arg(Pbf)-OH、Fmoc-(o-Boc)Dbz-OH、Fmoc-D-Tyr(tBu)-OH的顺序缩合后,20个残基通过微波辅助Fmoc-SPPS,然后通过标准Fmoc-SPPS延伸肽链。在TFA/H2O/m-甲酚/硫代茴香醚/EDT(80:5:5:5:5)中进行2小时的脱保护和与树脂的切割。通过过滤除去树脂后,使粗品沉淀,用冷Et2O洗涤。将得到的沉淀物溶解在含有0.1%TFA的50%CH3CN溶液中后,通过HPLC纯化粗品,得到所需的肽D-13(307mg、产率5.5%:序列号67)。
MS(ESI):C241H374N72O75S3的计算值:5576.25;测定值(m/z):[M+6H]6+=930.51、[M+5H]5+=1116.49、[M+4H]4+=1394.47、[M+3H]3+=1859.18。
(17)D-14的合成
通过与上述L-14的合成相同的工艺,得到D-14(12mg、产率6.1%:序列号68)。
MS(ESI):C150H236N46O49S的计算值:3499.87;测定值(m/z):[M+3H]3+=1167.42、[M+2H]2+=1750.46。
(18)D-15的合成
通过与上述L-15的合成相同的工艺,得到D-15(139mg、产率45%:序列号69)。
MS(ESI):C379H582N114O112S5的计算值:8687.81;测定值(m/z):[M+10H]10+=869.77、[M+9H]9+=965.86、[M+8H]8+=1086.95、[M+7H]7+=1241.88、[M+6H]6+=1449.03、[M+5H]5+=1738.06。
(19)D-16的合成
通过与上述L-16的合成相同的工艺,从D-15(95mg、11μmol)得到D-16(71mg、产率75%:序列号70)。
MS(ESI):C379H582N114O112S4的计算值:8655.75;测定值(m/z):[M+11H]11+=787.75、[M+10H]10+=866.42、[M+9H]9+=962.74、[M+8H]8+=1082.79、[M+7H]7+=1237.29、[M+6H]6+=1443.20、[M+5H]5+=1731.76。
(20)D-17的合成
通过与上述L-17的合成相同的工艺,得到D-17(39mg、产率76%:序列号71)。
MS(ESI):C376H577N113O111S4的计算值:8584.67;测定值(m/z):[M+11H]11+=781.13、[M+10H]10+=859.26、[M+9H]9+=954.63、[M+8H]8+=1074.18、[M+7H]7+=1227.17、[M+6H]6+=1431.62、[M+5H]5+=1717.09。
(21)D-18的合成
通过与上述L-18的合成相同的工艺,得到D-18(39mg、产率70%:序列号72)。
MS(ESI):C465H727N139O141S4的计算值:10649.00;测定值(m/z):[M+13H]13+=819.55、[M+12H]12+=888.24、[M+11H]11+=968.96、[M+10H]10+=1065.32、[M+9H]9+=1184.22、[M+8H]8+=1331.80、[M+7H]7+=1522.39、[M+6H]6+=1775.59。
(22)D-19的合成
通过与上述L-19的合成相同的工艺,得到D-19(10mg、产率58%:序列号73)。
MS(ESI):C460H714N132O141S5的计算值:10509.86;测定值(m/z):[M+11H]11+=955.88、[M+10H]10+=1051.94、[M+9H]9+=1168.23、[M+8H]8+=1314.67、[M+7H]7+=1502.28、[M+6H]6+=1751.97。
(23)D-20(D-PMP12A2h1)的合成
通过与上述L-20的合成相同的工艺,得到D-20(2.6mg、产率40%:序列号74)。
MS(ESI):C602H942N178O188S5的计算值:13841.52;测定值(m/z):[M+15H]15+=923.72、[M+14H]14+=989.70、[M+13H]13+=1065.83、[M+12H]12+=1154.30、[M+11H]11+=1259.49、[M+10H]10+=1384.91、[M+9H]9+=1539.04、[M+8H]8+=1731.03。
(24)PMP12A2h1及D-PMP12A2h1的折叠
使肽L-20或D-20溶解在含有6M胍和40mM DTT的PBS(pH 7.4)中至1.0mg/mL,室温孵育2小时。将该溶液用PBS(pH 7.4)稀释100倍,室温放置过夜。使用MWCO3000离心滤膜(Millipore、Amicon-Ultra3kDa)浓缩溶液。接着,添加10倍量的5,5’-二硫代双(2-苯甲酸)(DTNB),在37℃下孵育5小时。使用MWCO3000离心滤膜纯化目标蛋白。
(25)PMP12A2h1及D-PMP12A2h1的CD光谱
用PBS(pH 7.4)稀释折叠后的PMP12A2h1和D-PMP12A2h1,将浓度分别调整为10μM。各蛋白质的CD光谱使用圆二色光谱仪(JACSO J-720)在20℃下记录。
PMP12A2h1的CD光谱是具有216nm的极小值和203nm的极大值的光谱,表明存在β-折叠结构(图10)。D-PMP12A2h1的CD光谱是PMP12A2h1的CD光谱的正负符号相反的光谱,这暗示了D-PMP12A2h1折叠成为与PMP12A2h1对掌的结构。
(26)PMP12A2h1的表面等离子共振(SPR)分析
使用Biacore X100(Cytiva)进行实验。将重组vWF A1结构域(U-Protein ExpressBV,V0002)固定在CM5芯片上,使其成为2500RU。将折叠后的PMP12A2h1用HBS-EP缓冲液稀释并进行添加。以HBS-EP缓冲液为运行缓冲液,1M NaCl为解离溶液,分析物的接触时间为3分钟,解离时间为7分钟,以流速30μL/min进行测定。进行3次分析,解离常数由稳态的结合算出。
表明化学合成的PMP12A2h1对于vWF A1结构域以足够的结合亲和力结合(图11)。这暗示合成PMP12A2h1具有合适的生物活性。
实施例3
使用CDR区随机化的T7噬菌体库并尝试筛选由靶分子特异性D-氨基酸组成的VHH抗体。
(1)噬菌体库
使用卡拉西单抗的VHH抗体(PMP12A2h1)的序列中只有CDR3的随机库或CDR1、CDR2、CDR3这3个区全部随机化的T7噬菌体库。此时,使用CDR3的随机氨基酸的长度为7、10、13、17、20-mer的库。
(2)靶分子
作为靶分子,选择在已报道的协议中可以化学合成的分子(例如细胞因子、趋化因子),用使用D-氨基酸合成的镜像型蛋白质。此时,镜像型蛋白质的N-末端或C-末端附加6×His等标签。
(3)噬菌体筛选
将靶镜像型蛋白质溶解在Tris缓冲液TBS(50mM Tris-HCl、pH 7.5、150mM NaCl)中至1~10μg/ml的浓度,对Nickel板(Thermo Scientific公司)室温包衣30分钟(100μl/孔)。然后,使用封闭剂(3% BSA或4% BlockAce)室温封闭2小时。用PBST洗涤孔3次后,添加上述噬菌体库100μl(109~1010pfu),室温孵育30分钟。用洗涤液(PBST)洗涤孔10次,除去未结合的噬菌体。然后,加入100μl的elution buffer(含有500mM imidazole的TBS),孵育2分钟左右,洗脱靶分子和与其特异性结合的噬菌体。向含有大肠杆菌(BLT5403)的10ml LB培养基(Nacalai Tesque)中加入洗脱液,在37℃下振荡(190r/min)1.5~2小时直至大肠杆菌溶菌。另外,将加入到孔中的噬菌体和回收的噬菌体与4ml top agar(0.75% LB琼脂培养基、Nacalai Tesque)一起撒在部分受体培养皿(As One)中,并计算形成的斑块数。根据该值算出结合效率(浓缩率)。通过反复进行3~4次以上的作业,确认每一回合的结合效率是否上升。
(4)序列确认
从确认浓缩的噬菌体溶菌级分取得克隆,通过Sanger测序分析(ABI公司3130Genetic analyzer)一个一个地取得VHH的随机序列。另外,使用新一代可编程控制器(illumina公司iSeq 100system),全面取得随机区的排列。在随机区中,以具有显示高同源性序列的VHH抗体作为合成的对象。
(5)靶向特异性D-VHH抗体的合成
根据本发明的程序合成具有被确定为对象的CDR序列的D-VHH抗体。使用Biacoresystem等分析与由原始L-氨基酸组成的靶分子的亲和力。
工业实用性
本发明的低分子抗体具有与天然型的抗体分子同样的特异性,同时免疫原性降低。因此,在药物或诊断成像等医疗领域是有用的。
本说明书中引用的所有现有技术文献作为参考并入本文。
序列表自由文本
序列号55-合成肽L-1/D-1
序列号56-合成肽L-2/D-2
序列号57-合成肽L-3/D-3
序列号58-合成肽L-4/D-4
序列号59-合成肽L-5/D-5
序列号60-合成肽L-6/D-6
序列号61-合成肽L-7/D-7
序列号62-合成肽L-8/D-8
序列号63-合成肽L-9/D-9
序列号64-合成肽L-10/D-10
序列号65-合成肽L-11/D-11
序列号66-合成肽L-12/D-12
序列号67-合成肽L-13/D-13
序列号68-合成肽L-14/D-14
序列号69-合成肽L-15/D-15
序列号70-合成肽L-16/D-16
序列号71-合成肽L-17/D-17
序列号72-合成肽L-18/D-18
序列号73-合成肽L-19/D-19
序列号74-合成肽L-20/D-20
序列表
<110> 国立大学法人京都大学
<120> 免疫原性降低型低分子抗体
<130> PKD-9004WO
<150> JP2020-105104
<151> 2020-06-18
<160> 74
<170> PatentIn 3.5版本
<210> 1
<211> 25
<212> PRT
<213> 单峰驼
<400> 1
Gln Val Gln Leu Val Glu Ser Gly Gly Ala Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser
20 25
<210> 2
<211> 14
<212> PRT
<213> 单峰驼
<400> 2
Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val Ala
1 5 10
<210> 3
<211> 38
<212> PRT
<213> 单峰驼
<400> 3
Tyr Glu Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala
1 5 10 15
Arg Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr
20 25 30
Ala Val Tyr Tyr Cys Asn
35
<210> 4
<211> 11
<212> PRT
<213> 单峰驼
<400> 4
Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
1 5 10
<210> 5
<211> 25
<212> PRT
<213> 大羊驼
<400> 5
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ala
20 25
<210> 6
<211> 14
<212> PRT
<213> 大羊驼
<400> 6
Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val Gly
1 5 10
<210> 7
<211> 38
<212> PRT
<213> 大羊驼
<400> 7
Tyr Ala Asp Ser Val Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ala
1 5 10 15
Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr
20 25 30
Ala Val Tyr Tyr Cys Ala
35
<210> 8
<211> 11
<212> PRT
<213> 大羊驼
<400> 8
Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
1 5 10
<210> 9
<211> 25
<212> PRT
<213> 羊驼
<400> 9
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser
20 25
<210> 10
<211> 14
<212> PRT
<213> 羊驼
<400> 10
Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val Ala
1 5 10
<210> 11
<211> 36
<212> PRT
<213> 羊驼
<400> 11
Asp Ser Val Lys Gly Arg Tyr Thr Ile Ser Arg Asp Tyr Ala Lys Asn
1 5 10 15
Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Leu
20 25 30
Tyr Tyr Cys Asn
35
<210> 12
<211> 11
<212> PRT
<213> 羊驼
<400> 12
Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
1 5 10
<210> 13
<211> 25
<212> PRT
<213> 羊驼
<400> 13
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser
20 25
<210> 14
<211> 14
<212> PRT
<213> 羊驼
<400> 14
Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val Ala
1 5 10
<210> 15
<211> 38
<212> PRT
<213> 羊驼
<400> 15
Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala
1 5 10 15
Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr
20 25 30
Ala Val Tyr Tyr Cys Ala
35
<210> 16
<211> 11
<212> PRT
<213> 羊驼
<400> 16
Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
1 5 10
<210> 17
<211> 25
<212> PRT
<213> 单峰驼
<400> 17
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser
20 25
<210> 18
<211> 14
<212> PRT
<213> 单峰驼
<400> 18
Trp Tyr His Gln Ala Pro Gly Asn Glu Cys Glu Leu Val Ser
1 5 10
<210> 19
<211> 38
<212> PRT
<213> 单峰驼
<400> 19
Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala
1 5 10 15
Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr
20 25 30
Ala Met Tyr Tyr Cys Ala
35
<210> 20
<211> 11
<212> PRT
<213> 单峰驼
<400> 20
Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
1 5 10
<210> 21
<211> 128
<212> PRT
<213> 单峰驼
<400> 21
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Ser Val Gln Thr Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Glu Tyr Thr Gln Ser Ser Ala
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Gly Ile Ser Arg Phe Phe Gly Thr Ala Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Asp Lys Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Tyr Cys
85 90 95
Ala Ala Gly Gln Gly Cys Leu Thr Thr Ile Gln Ala Leu Gly Gly Ala
100 105 110
Tyr Gly Tyr Asn Ala Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 22
<211> 128
<212> PRT
<213> 骆驼科
<400> 22
Gln Val Gln Leu Gln 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 Lys Met Ser Ser Arg Arg
20 25 30
Cys Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Arg Val
35 40 45
Ala Lys Leu Leu Thr Thr Ser Gly Ser Thr Tyr Leu Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Asn Asn Ala Lys Ser Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Asp Ser Phe Glu Asp Pro Thr Cys Thr Leu Val Thr Ser Ser
100 105 110
Gly Ala Phe Gln Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 23
<211> 125
<212> PRT
<213> 大羊驼
<400> 23
Gln Val Gln Leu Gln 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 Phe Thr Leu Asp Tyr Tyr
20 25 30
Tyr Ile Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ser Cys Ile Ser Ser Ser His Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Thr Val Ala Val Ala His Phe Arg Gly Cys Gly Val Asp Gly Met
100 105 110
Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 24
<211> 115
<212> PRT
<213> 大羊驼
<400> 24
Gln Val Gln Leu Val Glu Ser Gly Gly Ala Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Pro Val Asn Arg Tyr
20 25 30
Ser Met Arg Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val
35 40 45
Ala Gly Met Ser Ser Ala Gly Asp Arg Ser Ser Tyr Glu Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Arg Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Asn Val Asn Val Gly Phe Glu Tyr Trp Gly Gln Gly Thr Gln Val Thr
100 105 110
Val Ser Ser
115
<210> 25
<211> 128
<212> PRT
<213> 人工序列
<220>
<223> 人化VHH (人-美洲驼)
<400> 25
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 Arg Thr Phe Ser Tyr Asn
20 25 30
Pro Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Arg Glu Leu Val
35 40 45
Ala Ala Ile Ser Arg Thr Gly Gly Ser Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Glu Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Arg Met Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Ala Gly Val Arg Ala Glu Asp Gly Arg Val Arg Thr Leu Pro
100 105 110
Ser Glu Tyr Thr Phe Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 26
<211> 125
<212> PRT
<213> 大羊驼
<400> 26
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Glu Tyr
20 25 30
Ala Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val
35 40 45
Ala Thr Ile Ser Trp Ser Gly Gly Ser Thr Tyr Tyr Thr Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Ala Gly Leu Gly Thr Val Val Ser Glu Trp Asp Tyr Asp Tyr
100 105 110
Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 27
<211> 120
<212> PRT
<213> 大羊驼
<400> 27
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Ser Ile Phe Ser Ile Asn
20 25 30
Ala Met Asp Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val
35 40 45
Ala Gly Ile Thr Ser Gly Gly Ser Thr Asn Tyr Gly Asp Phe Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu
65 70 75 80
Gln Met Asp Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Ala Glu Val Gly Gly Trp Gly Pro Pro Arg Pro Asp Tyr Trp Gly His
100 105 110
Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 28
<211> 123
<212> PRT
<213> 单峰驼
<400> 28
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Leu Ser Thr Tyr
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu His Glu Gly Ala
35 40 45
Ala Phe Ile Asn Ser Gly Gly Gly Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Leu
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Tyr Cys
85 90 95
Ala Ala Gly Arg Tyr Cys Ile Gly Gly Tyr Pro Gly Gly Gly Thr Tyr
100 105 110
Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 29
<211> 117
<212> PRT
<213> 单峰驼
<400> 29
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Gly Val Asp Gly Thr
20 25 30
Asp Met Gly Trp Tyr Arg Gln Ala Pro Gly Asn Glu Cys Glu Leu Val
35 40 45
Ser Ser Ile Ser Ser Ile Gly Ile Gly Tyr Tyr Ser Glu Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Pro Asp Asp Thr Ala Val Tyr Tyr Cys Gly
85 90 95
Arg Arg Trp Ile Gly Tyr Arg Cys Gly Asn Trp Gly Arg Gly Thr Gln
100 105 110
Val Thr Val Ser Ser
115
<210> 30
<211> 118
<212> PRT
<213> 单峰驼
<400> 30
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ile Thr Phe Ser Ile Asn
20 25 30
Thr Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val
35 40 45
Ala Leu Ile Ser Ser Ile Gly Asp Thr Tyr Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Lys
85 90 95
Arg Phe Arg Thr Ala Ala Gln Gly Thr Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Gln Val Thr Val Ser Ser
115
<210> 31
<211> 122
<212> PRT
<213> 大羊驼
<400> 31
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ala Gly Arg Asn Leu Arg Met Tyr
20 25 30
Arg Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val
35 40 45
Gly Thr Met Val Trp Ser Ser Asp Thr Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Gly Ala Gly Trp Ala Gly Thr Met Thr Asp Tyr Asn Tyr Trp
100 105 110
Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 32
<211> 127
<212> PRT
<213> 大羊驼
<400> 32
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Thr Ser Phe
20 25 30
Ala Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val
35 40 45
Ala Ser Ile Ser Arg Ser Gly Thr Leu Thr Arg Tyr Ala Asp Ser Ala
50 55 60
Lys Gly Arg Phe Thr Ile Ser Val Asp Asn Ala Lys Asn Thr Val Ser
65 70 75 80
Leu Gln Met Asp Asn Leu Asn Pro Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Asp Leu His Arg Pro Tyr Gly Pro Gly Thr Gln Arg Ser Asp
100 105 110
Glu Tyr Asp Ser Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 33
<211> 127
<212> PRT
<213> 大羊驼
<400> 33
Gln Val Gln Leu Gln 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 Arg Thr Phe Ser Asn Tyr
20 25 30
Arg Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val
35 40 45
Ala Thr Ile Ser Gln Ser Gly Ala Ala Thr Ala Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Phe Ser Arg Asp Asn Ala Lys Asn Leu Leu Tyr
65 70 75 80
Leu Glu Met Leu Ser Leu Glu Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Ser Ser Arg Val Phe Tyr Thr Glu Val Leu Gln Thr Thr Thr
100 105 110
Gly Tyr Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 34
<211> 125
<212> PRT
<213> 大羊驼
<400> 34
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Asn Asp
20 25 30
His Met Gly Trp Phe Arg Lys Ala Pro Gly Lys Glu Arg Glu Phe Val
35 40 45
Ala Ala Ile Thr Pro Gly Thr Glu Lys Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ala Phe Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Ala Thr Pro Tyr Tyr Arg Gly Ser Tyr Tyr Ala Ala Ser Thr Tyr
100 105 110
Thr Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 35
<211> 117
<212> PRT
<213> 羊驼
<400> 35
Gln Val Gln Leu Ala Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ser Ala Ser Gly Gly Val Phe Ile Ile Tyr
20 25 30
Asn Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val
35 40 45
Ala Ser Ile Asp Ser Tyr Ser Gly Ser Ile Thr Asn Tyr Ala Asp Ser
50 55 60
Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Val Glu Lys Arg Val
65 70 75 80
Tyr Leu Glu Met Asn Asn Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr
85 90 95
Cys Asn Ala Asn Leu Arg Thr Asn Asn Tyr Trp Gly Gln Gly Thr Gln
100 105 110
Val Thr Val Ser Ser
115
<210> 36
<211> 118
<212> PRT
<213> 羊驼
<400> 36
Gln Val Gln Leu Gln 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 Ser Ile Thr Ser Thr Asn
20 25 30
Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val
35 40 45
Ala Gln Ile Thr Asn Tyr Gly Ala Ser Asn Tyr Gly Asp Ser Val Lys
50 55 60
Gly Arg Tyr Thr Ile Ser Arg Asp Tyr Ala Lys Asn Thr Val Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Leu Tyr Tyr Cys Asn
85 90 95
Val Arg Glu Tyr Arg Phe Glu Val Ala Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Gln Val Thr Val Ser Ser
115
<210> 37
<211> 130
<212> PRT
<213> 羊驼
<400> 37
Gln Val Gln Leu Gln 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 Phe Thr Phe Asp Tyr Tyr
20 25 30
Ala Val Gly Trp Phe Arg His Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ser Cys Ile Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Val Lys Gly
50 55 60
Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln
65 70 75 80
Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Tyr Cys Ala Ala
85 90 95
Ile Lys Arg Gly Phe Cys Thr Ser Trp Gly Gly Glu Val Pro Ala Pro
100 105 110
Lys Ser Ala Asp Tyr Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val
115 120 125
Ser Ser
130
<210> 38
<211> 122
<212> PRT
<213> 羊驼
<400> 38
Gln Val Gln Leu Gln 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 Phe Thr Phe Ser Arg Tyr
20 25 30
Pro Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Ala Ile Asn Ser Ser Gly Glu Ala Thr Leu Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Thr Gly Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Asp Asp Gly Lys Leu Leu Pro Gln Ser Thr Thr Thr Trp Tyr Arg
100 105 110
Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 39
<211> 124
<212> PRT
<213> 羊驼
<400> 39
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Thr Leu Ser Cys Thr Arg Thr Gly Phe Thr Ala Ser Thr Asn
20 25 30
Ala Tyr Gly Trp Tyr Arg Gln Gly Pro Gly Lys Lys Cys Glu Trp Val
35 40 45
Ser Tyr Met Thr Ile Pro Ser Gly Arg Thr Thr Tyr Ala Asp Ala Val
50 55 60
Lys Gly Arg Phe Ala Met Ser Arg Asp Lys Ala Lys Ser Thr Val Leu
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Gly Asp Val Pro Phe Ser Thr Leu Pro Ala Met Cys Thr Asn Asp Gly
100 105 110
Pro Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 40
<211> 124
<212> PRT
<213> 羊驼
<400> 40
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Thr Leu Ser Cys Thr Arg Thr Gly Leu Thr Pro Ser Thr Gly
20 25 30
Ala Tyr Gly Trp Tyr Arg Gln Ala Pro Gly Lys Lys Cys Glu Leu Val
35 40 45
Ser Tyr Ile Thr Ile Pro Ser Gly Arg Thr Thr Tyr Thr Asp Ser Val
50 55 60
Lys Gly Arg Phe Ala Ile Ser Arg Asp Lys Ala Lys Asn Thr Val Phe
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Leu Tyr Tyr Cys
85 90 95
Gly Asp Val Pro Tyr Ser Thr Ile Gln Ala Met Cys Thr Asp Asp Gly
100 105 110
Pro Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 41
<211> 124
<212> PRT
<213> 羊驼
<400> 41
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Thr Leu Ser Cys Thr Arg Thr Gly Leu Thr Ala Ser Thr Asn
20 25 30
Ala Tyr Gly Trp Tyr Arg Gln Ala Pro Gly Lys Lys Cys Glu Leu Val
35 40 45
Ser Tyr Ile Thr Ile Pro Ser Gly Arg Thr Thr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ala Ile Ser Arg Asp Lys Ala Lys Asn Thr Val Phe
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Leu Tyr Tyr Cys
85 90 95
Gly Asp Val Pro Tyr Ser Thr Ile Gln Ala Met Cys Thr Asp Asp Gly
100 105 110
Pro Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 42
<211> 114
<212> PRT
<213> 羊驼
<400> 42
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ser Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Arg Ala Ser Gly Phe Thr Phe Ser His Tyr
20 25 30
Pro Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Lys Arg Glu Leu Val
35 40 45
Ala Gly Ile Tyr Thr Asp Gly Ser Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu
65 70 75 80
Gln Met Ser Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Arg
85 90 95
Arg Ser Met Val Ser Val Tyr Trp Gly Gln Gly Thr Gln Val Thr Val
100 105 110
Ser Ser
<210> 43
<211> 120
<212> PRT
<213> 羊驼
<400> 43
Gln Val Gln Leu Gln 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 Arg Thr Phe Ser Tyr Tyr
20 25 30
Thr Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val
35 40 45
Ala Ser Ile Ser Trp Ser Gly Gly Ile Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Arg Lys Glu Pro Gly Ser Ala Tyr Phe Gly Ser Trp Gly Gln
100 105 110
Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 44
<211> 124
<212> PRT
<213> 羊驼
<400> 44
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Thr Leu Ser Cys Thr Arg Thr Gly Phe Thr Ala Ser Thr Asn
20 25 30
Ala Tyr Gly Trp Tyr Arg Gln Ala Pro Gly Lys Lys Cys Glu Leu Val
35 40 45
Ser Tyr Ile Thr Ile Pro Ser Gly Arg Thr Thr Tyr Ala Asp Ala Val
50 55 60
Lys Gly Arg Phe Ala Ile Ser Arg Asp Lys Ala Lys Asn Thr Val Phe
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Gly Asp Val Pro Phe Ser Thr Leu Pro Ala Met Cys Thr Asn Asp Gly
100 105 110
Pro Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 45
<211> 124
<212> PRT
<213> 羊驼
<400> 45
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Thr Leu Ser Cys Thr Arg Thr Gly Phe Thr Ala Ser Thr Asn
20 25 30
Ala Tyr Gly Trp Tyr Arg Gln Gly Pro Gly Lys Lys Cys Glu Trp Val
35 40 45
Ser Tyr Met Thr Ile Pro Ser Gly Arg Thr Thr Tyr Ala Asp Ala Val
50 55 60
Lys Gly Arg Phe Ala Met Ser Arg Asp Lys Ala Lys Ser Thr Val Phe
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Gly Asp Val Pro Phe Ser Thr Leu Pro Ala Met Cys Thr Asn Asp Gly
100 105 110
Pro Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 46
<211> 126
<212> PRT
<213> 单峰驼
<400> 46
Gln Val Gln Leu Gln 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 Tyr Pro Tyr Gly Tyr Thr
20 25 30
Phe Ser Ser Tyr Cys Met Arg Trp Phe Arg Gln Ala Pro Gly Lys Asp
35 40 45
Arg Glu Gly Val Ala Arg Phe Glu Arg Asn Gly Leu Thr Thr Tyr Tyr
50 55 60
Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Val Lys
65 70 75 80
Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala
85 90 95
Thr Tyr Tyr Cys Ala Ala Ala Pro Lys Gln Leu Arg Thr Cys Gly Asp
100 105 110
Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 47
<211> 131
<212> PRT
<213> 单峰驼
<400> 47
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Ser Val Gln Val Gly Gly
1 5 10 15
Ser Leu Arg Val Ala Cys Ala Ala Ser Gly Asp Thr Phe Ser Gly Tyr
20 25 30
Leu Ala Ala Trp Phe Arg Gln Ala Pro Gly Lys Gly Arg Glu Gly Val
35 40 45
Ala Ala Ile Asn Ser Lys Arg His Thr Thr Ser Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Lys Asp Asn Ala Asp Asn Ile Met Tyr
65 70 75 80
Leu Glu Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Tyr Cys
85 90 95
Ala Ala Ala Asp Ala Ile Gly Leu Ala Glu Tyr Trp Ser Thr Pro Thr
100 105 110
Leu Ser Ala Ala Arg Tyr Lys Tyr Trp Gly Gln Gly Thr Gln Val Thr
115 120 125
Val Ser Ser
130
<210> 48
<211> 129
<212> PRT
<213> 单峰驼
<400> 48
Asp Val Gln Leu Gln Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Pro Tyr Phe Ser Leu Arg Ser
20 25 30
Cys Val Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Ala Ile Phe Thr Ser Thr Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Asn Thr Val Ser Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Ala Ala Met Tyr Tyr Cys Ala
85 90 95
Ile Glu Gly Arg Pro Met Ile Gly Gly Pro Ser Cys Ser Met Gly Ser
100 105 110
Pro Asn Ser Tyr Thr Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser
115 120 125
Ser
<210> 49
<211> 127
<212> PRT
<213> 单峰驼
<400> 49
Asp Val Gln Leu Gln Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ser Val Ser Gly Tyr Thr Phe Ser Met Leu
20 25 30
Ala Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val
35 40 45
Ala Val Ile Ser Pro Gly Ser Gly Phe Thr Tyr Tyr Asp Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Arg Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ser Ala Gly Leu Trp Ser Glu Gly Ile Ser Asn Val Arg Ala Pro Asp
100 105 110
Gly Tyr Lys Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 50
<211> 116
<212> PRT
<213> 单峰驼
<400> 50
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Thr Ser Gly Ser Ile Phe Ser Ile Ile
20 25 30
Ser Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val
35 40 45
Ala Leu Val Phe Arg Gly Gly Ser Thr Val Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Gly Asp Ile Ala Lys Ser Thr Val Tyr Leu
65 70 75 80
Gln Met Asp Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn
85 90 95
Ala Lys Pro Ile Gly Thr Ala Gln Tyr Trp Gly Gln Gly Thr Gln Val
100 105 110
Thr Val Ser Ser
115
<210> 51
<211> 125
<212> PRT
<213> 单峰驼
<400> 51
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Asp Asp Ser
20 25 30
Asp Met Gly Trp Tyr His Gln Ala Pro Gly Asn Glu Cys Glu Leu Val
35 40 45
Ser Ala Ile Phe Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala
85 90 95
Ala Ala Thr Thr Thr Val Ala Ser Pro Pro Val Arg His Val Cys Asn
100 105 110
Gly Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 52
<211> 122
<212> PRT
<213> 单峰驼
<220>
<221> MOD_RES
<222> (116)..(122)
<223> 具有His-标签的赖氨酸
<400> 52
Gln Val Gln Leu Val Glu Ser Gly Gly Ala Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Pro Val Asn Arg Tyr
20 25 30
Ser Met Arg Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val
35 40 45
Ala Gly Met Ser Ser Ala Gly Asp Arg Ser Ser Tyr Glu Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Arg Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Asn Val Asn Val Gly Phe Glu Tyr Trp Gly Gln Gly Thr Gln Val Thr
100 105 110
Val Ser Ser Lys His His His His His His
115 120
<210> 53
<211> 126
<212> PRT
<213> 骆驼科
<220>
<221> MOD_RES
<222> (121)..(126)
<223> His标签
<220>
<221> MOD_RES
<222> (121)..(126)
<223> His-标签
<400> 53
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Thr Ser Gly Ile Thr Phe Met Arg Tyr
20 25 30
Ala Leu Gly Trp Tyr Arg Gln Ser Pro Gly Lys Gln Arg Glu Met Val
35 40 45
Ala Ser Ile Asn Ser Gly Gly Thr Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn
85 90 95
Ala Arg Trp Val Lys Pro Gln Phe Ile Asp Asn Asn Tyr Trp Gly Gln
100 105 110
Gly Thr Gln Val Thr Val Ser Ser His His His His His His
115 120 125
<210> 54
<211> 128
<212> PRT
<213> 人工序列
<220>
<223> 人化VHH (人-美洲驼)
<400> 54
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 Arg Thr Phe Ser Tyr Asn
20 25 30
Pro Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Arg Glu Leu Val
35 40 45
Ala Ala Ile Ser Arg Thr Gly Gly Ser Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Glu Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Arg Met Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Ala Gly Val Arg Ala Glu Asp Gly Arg Val Arg Thr Leu Pro
100 105 110
Ser Glu Tyr Thr Phe Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 55
<211> 21
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-1/D-1
<400> 55
Gln Val Gln Leu Val Glu Ser Gly Gly Ala Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser
20
<210> 56
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-2/D-2
<400> 56
Cys Ala Ala Ser Gly Phe Pro Val Asn Arg Tyr Ser Met Arg Trp Tyr
1 5 10 15
Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val Ala Gly Met Ser Ser
20 25 30
<210> 57
<211> 42
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-3/D-3
<400> 57
Cys Gly Asp Arg Ser Ser Tyr Glu Asp Ser Val Lys Gly Arg Phe Thr
1 5 10 15
Ile Ser Arg Asp Asp Ala Arg Asn Thr Val Tyr Leu Gln Met Asn Ser
20 25 30
Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr
35 40
<210> 58
<211> 27
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-4/D-4
<400> 58
Cys Asn Val Asn Val Gly Phe Glu Tyr Trp Gly Gln Gly Thr Gln Val
1 5 10 15
Thr Val Ser Ser Lys His His His His His His
20 25
<210> 59
<211> 74
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-5/D-5
<400> 59
Cys Ala Ala Ser Gly Phe Pro Val Asn Arg Tyr Ser Met Arg Trp Tyr
1 5 10 15
Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val Ala Gly Met Ser Ser
20 25 30
Cys Gly Asp Arg Ser Ser Tyr Glu Asp Ser Val Lys Gly Arg Phe Thr
35 40 45
Ile Ser Arg Asp Asp Ala Arg Asn Thr Val Tyr Leu Gln Met Asn Ser
50 55 60
Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr
65 70
<210> 60
<211> 74
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-6/D-6
<400> 60
Cys Ala Ala Ser Gly Phe Pro Val Asn Arg Tyr Ser Met Arg Trp Tyr
1 5 10 15
Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val Ala Gly Met Ser Ser
20 25 30
Ala Gly Asp Arg Ser Ser Tyr Glu Asp Ser Val Lys Gly Arg Phe Thr
35 40 45
Ile Ser Arg Asp Asp Ala Arg Asn Thr Val Tyr Leu Gln Met Asn Ser
50 55 60
Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr
65 70
<210> 61
<211> 74
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-7/D-7
<400> 61
Cys Ala Ala Ser Gly Phe Pro Val Asn Arg Tyr Ser Met Arg Trp Tyr
1 5 10 15
Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val Ala Gly Met Ser Ser
20 25 30
Ala Gly Asp Arg Ser Ser Tyr Glu Asp Ser Val Lys Gly Arg Phe Thr
35 40 45
Ile Ser Arg Asp Asp Ala Arg Asn Thr Val Tyr Leu Gln Met Asn Ser
50 55 60
Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr
65 70
<210> 62
<211> 95
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-8/D-8
<400> 62
Gln Val Gln Leu Val Glu Ser Gly Gly Ala Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Pro Val Asn Arg Tyr
20 25 30
Ser Met Arg Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val
35 40 45
Ala Gly Met Ser Ser Ala Gly Asp Arg Ser Ser Tyr Glu Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Arg Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr
85 90 95
<210> 63
<211> 95
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-9/D-9
<400> 63
Gln Val Gln Leu Val Glu Ser Gly Gly Ala Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Pro Val Asn Arg Tyr
20 25 30
Ser Met Arg Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val
35 40 45
Ala Gly Met Ser Ser Ala Gly Asp Arg Ser Ser Tyr Glu Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Arg Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr
85 90 95
<210> 64
<211> 122
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-10/D-10
<400> 64
Gln Val Gln Leu Val Glu Ser Gly Gly Ala Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Pro Val Asn Arg Tyr
20 25 30
Ser Met Arg Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val
35 40 45
Ala Gly Met Ser Ser Ala Gly Asp Arg Ser Ser Tyr Glu Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Arg Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Asn Val Asn Val Gly Phe Glu Tyr Trp Gly Gln Gly Thr Gln Val Thr
100 105 110
Val Ser Ser Lys His His His His His His
115 120
<210> 65
<211> 21
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-11/D-11
<400> 65
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
20
<210> 66
<211> 28
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-12/D-12
<400> 66
Cys Ala Ala Ser Gly Arg Thr Phe Ser Tyr Asn Pro Met Gly Trp Phe
1 5 10 15
Arg Gln Ala Pro Gly Lys Gly Arg Glu Leu Val Ala
20 25
<210> 67
<211> 46
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-13/D-13
<400> 67
Cys Ile Ser Arg Thr Gly Gly Ser Thr Tyr Tyr Pro Asp Ser Val Glu
1 5 10 15
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Arg Met Val Tyr Leu
20 25 30
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr
35 40 45
<210> 68
<211> 33
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-14/D-14
<400> 68
Cys Ala Ala Ala Gly Val Arg Ala Glu Asp Gly Arg Val Arg Thr Leu
1 5 10 15
Pro Ser Glu Tyr Thr Phe Trp Gly Gln Gly Thr Gln Val Thr Val Ser
20 25 30
Ser
<210> 69
<211> 74
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-15/D-15
<400> 69
Cys Ala Ala Ser Gly Arg Thr Phe Ser Tyr Asn Pro Met Gly Trp Phe
1 5 10 15
Arg Gln Ala Pro Gly Lys Gly Arg Glu Leu Val Ala Cys Ile Ser Arg
20 25 30
Thr Gly Gly Ser Thr Tyr Tyr Pro Asp Ser Val Glu Gly Arg Phe Thr
35 40 45
Ile Ser Arg Asp Asn Ala Lys Arg Met Val Tyr Leu Gln Met Asn Ser
50 55 60
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr
65 70
<210> 70
<211> 74
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-16/D-16
<400> 70
Cys Ala Ala Ser Gly Arg Thr Phe Ser Tyr Asn Pro Met Gly Trp Phe
1 5 10 15
Arg Gln Ala Pro Gly Lys Gly Arg Glu Leu Val Ala Ala Ile Ser Arg
20 25 30
Thr Gly Gly Ser Thr Tyr Tyr Pro Asp Ser Val Glu Gly Arg Phe Thr
35 40 45
Ile Ser Arg Asp Asn Ala Lys Arg Met Val Tyr Leu Gln Met Asn Ser
50 55 60
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr
65 70
<210> 71
<211> 74
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-17/D-17
<400> 71
Cys Ala Ala Ser Gly Arg Thr Phe Ser Tyr Asn Pro Met Gly Trp Phe
1 5 10 15
Arg Gln Ala Pro Gly Lys Gly Arg Glu Leu Val Ala Ala Ile Ser Arg
20 25 30
Thr Gly Gly Ser Thr Tyr Tyr Pro Asp Ser Val Glu Gly Arg Phe Thr
35 40 45
Ile Ser Arg Asp Asn Ala Lys Arg Met Val Tyr Leu Gln Met Asn Ser
50 55 60
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr
65 70
<210> 72
<211> 95
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-18/D-18
<400> 72
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 Arg Thr Phe Ser Tyr Asn
20 25 30
Pro Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Arg Glu Leu Val
35 40 45
Ala Ala Ile Ser Arg Thr Gly Gly Ser Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Glu Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Arg Met Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr
85 90 95
<210> 73
<211> 95
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-19/D-19
<400> 73
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 Arg Thr Phe Ser Tyr Asn
20 25 30
Pro Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Arg Glu Leu Val
35 40 45
Ala Ala Ile Ser Arg Thr Gly Gly Ser Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Glu Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Arg Met Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr
85 90 95
<210> 74
<211> 128
<212> PRT
<213> 人工序列
<220>
<223> 合成肽 L-20/D-20
<400> 74
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 Arg Thr Phe Ser Tyr Asn
20 25 30
Pro Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Arg Glu Leu Val
35 40 45
Ala Ala Ile Ser Arg Thr Gly Gly Ser Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Glu Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Arg Met Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Ala Gly Val Arg Ala Glu Asp Gly Arg Val Arg Thr Leu Pro
100 105 110
Ser Glu Tyr Thr Phe Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
Claims (14)
1.一种低分子抗体,所述低分子抗体由D-氨基酸和非手性甘氨酸构成,并且所述低分子抗体的分子量为60KDa及以下。
2.根据权利要求1所述的低分子抗体,所述低分子抗体是选自单一结构域抗体、scFV和Fab中的任一种。
3.根据权利要求1所述的低分子抗体,所述低分子抗体是VHH抗体。
4.根据权利要求1所述的低分子抗体,所述低分子抗体是具有:
(1)由序列号1~4表示的氨基酸序列或由具有与所述序列至少60%的同一性的氨基酸序列分别表示的4个骨架区;
(2)由序列号5~8表示的氨基酸序列或由具有与所述序列至少60%的同一性的氨基酸序列分别表示的4个骨架区;
(3)由序列号9~12表示的氨基酸序列或由具有与所述序列至少60%的同一性的氨基酸序列分别表示的4个骨架区;
(4)由序列号13~16表示的氨基酸序列或由具有与所述序列至少60%的同一性的氨基酸序列分别表示的4个骨架区;或者
(5)由序列号17~20表示的氨基酸序列或由具有与所述序列至少60%的同一性的氨基酸序列分别表示的4个骨架区;的VHH抗体。
5.一种由D-氨基酸和非手性甘氨酸构成的低分子抗体的制造方法,所述方法包括:
1)将低分子抗体的氨基酸序列分割为以半胱氨酸残基或丙氨酸残基为N末端的多个片段;
2)通过固相法化学合成各片段;以及
3)通过NCL法将合成的各片段依次连结,合成由D-氨基酸和非手性甘氨酸构成的低分子抗体。
6.根据权利要求5所述的方法,其中,所述低分子抗体是选自单一结构域抗体、scFV和Fab中的任一种。
7.根据权利要求5所述的方法,其中,所述低分子抗体是VHH抗体。
8.一种针对靶蛋白的、由D-氨基酸和非手性甘氨酸构成的低分子抗体的制造方法,所述方法包括:
1)提供针对靶蛋白的、由D-氨基酸和非手性甘氨酸构成的镜像型靶蛋白;
2)使用所述镜像型靶蛋白筛选抗体库,取得对所述镜像型靶蛋白具有亲和力的抗体;以及
3)基于得到的抗体的氨基酸序列,合成由D-氨基酸和非手性甘氨酸构成的低分子抗体。
9.根据权利要求8所述的方法,其中,由D-氨基酸和非手性甘氨酸构成的低分子抗体的合成包括以下工序:
1)将低分子抗体的氨基酸序列分割为以半胱氨酸残基或丙氨酸残基为N末端的多个片段;
2)通过固相法化学合成各片段;以及
3)通过NCL法将合成的各片段依次连结,合成由D-氨基酸和非手性甘氨酸构成的低分子抗体。
10.根据权利要求8所述的方法,其中,所述低分子抗体是选自单一结构域抗体、scFV和Fab中的任一种。
11.根据权利要求8所述的方法,其中,所述低分子抗体是VHH抗体。
12.一种多价或多特异性抗体,所述多价或多特异性抗体通过连结权利要求1至4中任一项所述的低分子抗体而得到。
13.一种由D-氨基酸构成的多价或多特异性抗体的制造方法,所述制造方法包括连结用权利要求5至11中任一项所述的方法制造的低分子抗体。
14.一种由D-氨基酸构成的多价或多特异性抗体的制造用试剂盒,所述试剂盒包括分别包含下述(a)或(b)的氨基酸序列的、由D-氨基酸和非手性甘氨酸构成的一个或多个多肽:
(a)由序列号55~57、61及63中任一个表示的氨基酸序列;
(b)具有与由序列号55~57、61及63中任一个表示的氨基酸序列至少60%的同一性的氨基酸序列。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2020-105104 | 2020-06-18 | ||
JP2020105104 | 2020-06-18 | ||
PCT/JP2021/022968 WO2021256524A1 (ja) | 2020-06-18 | 2021-06-17 | 免疫原性低減型低分子抗体とその製造法 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN116096747A true CN116096747A (zh) | 2023-05-09 |
Family
ID=79268044
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202180056535.9A Pending CN116096747A (zh) | 2020-06-18 | 2021-06-17 | 免疫原性降低型低分子抗体及其制造法 |
Country Status (5)
Country | Link |
---|---|
US (1) | US20230227536A1 (zh) |
EP (1) | EP4169940A1 (zh) |
JP (1) | JPWO2021256524A1 (zh) |
CN (1) | CN116096747A (zh) |
WO (1) | WO2021256524A1 (zh) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20240093179A1 (en) * | 2022-02-17 | 2024-03-21 | Specifica, Inc. | Single domain antibody libraries with maximized antibody developability characteristics |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2016506720A (ja) * | 2013-01-21 | 2016-03-07 | アッヴィ・インコーポレイテッド | 炎症性疾患のための抗tnf及び抗il17併用治療薬バイオマーカー |
US20170107541A1 (en) * | 2014-06-17 | 2017-04-20 | Poseida Therapeutics, Inc. | A method for directing proteins to specific loci in the genome and uses thereof |
JP7224585B2 (ja) | 2018-12-27 | 2023-02-20 | 山田化学工業株式会社 | フォトクロミック化合物 |
-
2021
- 2021-06-17 CN CN202180056535.9A patent/CN116096747A/zh active Pending
- 2021-06-17 EP EP21826416.6A patent/EP4169940A1/en not_active Withdrawn
- 2021-06-17 WO PCT/JP2021/022968 patent/WO2021256524A1/ja unknown
- 2021-06-17 JP JP2022531894A patent/JPWO2021256524A1/ja active Pending
- 2021-06-17 US US18/001,946 patent/US20230227536A1/en active Pending
Also Published As
Publication number | Publication date |
---|---|
US20230227536A1 (en) | 2023-07-20 |
JPWO2021256524A1 (zh) | 2021-12-23 |
WO2021256524A1 (ja) | 2021-12-23 |
EP4169940A1 (en) | 2023-04-26 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP7221259B2 (ja) | 改変した抗体組成物、それを作製および使用する方法 | |
JP2020186248A5 (zh) | ||
JP6165713B2 (ja) | インスリン様増殖因子1に特異的に結合する抗体 | |
JP2022023181A (ja) | C末端リジンで結合体化された免疫グロブリン | |
US12043663B2 (en) | Anti-TNF-alpha-antibodies and functional fragments thereof | |
JP2005535301A5 (zh) | ||
CN106459216A (zh) | 多特异性抗体构建体 | |
JP2005528914A5 (zh) | ||
JP2019506140A5 (zh) | ||
JP2015535523A (ja) | アプロチニン由来ポリペプチド−抗体コンジュゲート | |
JP2020515277A5 (zh) | ||
CN116096747A (zh) | 免疫原性降低型低分子抗体及其制造法 | |
US11236386B2 (en) | Method for labeling of aldehyde containing target molecules | |
US11104737B2 (en) | ACVR2A-specific antibody and method of treatment of muscle atrophy | |
US20230265187A1 (en) | Anti-tigit antibody and methods of use thereof | |
US20240294654A1 (en) | Anti-gipr antibody and methods of use thereof | |
US20220298246A1 (en) | Anti-b7-h3 antibody and methods of use thereof | |
CN116322770A (zh) | 对粘蛋白-1特异的抗体及其使用方法 | |
EP4007604A1 (en) | Antibody specific for gpc3 and methods of use thereof | |
TW201102086A (en) | Antibodies against human CCN1 and uses thereof | |
AU2020299600A1 (en) | Anti-CD38 antibody and methods of use thereof | |
CN114846030A (zh) | 催化抗体38c2的新型缀合化学 | |
WO2024074762A1 (en) | Ultrastable antibody fragments with a novel disuldide bridge | |
JP2024534964A (ja) | 抗ang2抗体、その調製方法及び使用 |
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 |