US20020068303A1 - Antigenic polypeptide sequences of factor VIII, and fragments and/or epitopes of these sequences - Google Patents
Antigenic polypeptide sequences of factor VIII, and fragments and/or epitopes of these sequences Download PDFInfo
- Publication number
- US20020068303A1 US20020068303A1 US09/853,080 US85308001A US2002068303A1 US 20020068303 A1 US20020068303 A1 US 20020068303A1 US 85308001 A US85308001 A US 85308001A US 2002068303 A1 US2002068303 A1 US 2002068303A1
- Authority
- US
- United States
- Prior art keywords
- epitope
- ser
- asp
- inclusive
- seq
- 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.)
- Abandoned
Links
- 229960000301 factor viii Drugs 0.000 title claims abstract description 104
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 97
- 108010054218 Factor VIII Proteins 0.000 title claims abstract description 94
- 102000001690 Factor VIII Human genes 0.000 title claims abstract description 94
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 71
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 49
- 230000000890 antigenic effect Effects 0.000 title claims abstract description 43
- 239000012634 fragment Substances 0.000 title claims description 123
- 239000003112 inhibitor Substances 0.000 claims description 112
- 150000001413 amino acids Chemical group 0.000 claims description 91
- 229940024606 amino acid Drugs 0.000 claims description 89
- 235000001014 amino acid Nutrition 0.000 claims description 89
- 238000009739 binding Methods 0.000 claims description 37
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims description 36
- 235000003704 aspartic acid Nutrition 0.000 claims description 35
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims description 35
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims description 34
- 239000004475 Arginine Substances 0.000 claims description 33
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims description 30
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 30
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 claims description 29
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 claims description 29
- 238000000034 method Methods 0.000 claims description 27
- 235000002374 tyrosine Nutrition 0.000 claims description 26
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 25
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims description 25
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims description 24
- 235000004279 alanine Nutrition 0.000 claims description 24
- 238000004587 chromatography analysis Methods 0.000 claims description 20
- 108010016626 Dipeptides Proteins 0.000 claims description 17
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims description 17
- 235000013922 glutamic acid Nutrition 0.000 claims description 17
- 239000004220 glutamic acid Substances 0.000 claims description 17
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 15
- 208000026278 immune system disease Diseases 0.000 claims description 15
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 14
- 239000004471 Glycine Substances 0.000 claims description 12
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 claims description 12
- 239000004472 Lysine Substances 0.000 claims description 12
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 claims description 12
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 claims description 11
- 150000003904 phospholipids Chemical class 0.000 claims description 11
- 210000002966 serum Anatomy 0.000 claims description 11
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 claims description 10
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 claims description 10
- 229930182817 methionine Natural products 0.000 claims description 10
- 239000008194 pharmaceutical composition Substances 0.000 claims description 10
- 239000004474 valine Substances 0.000 claims description 10
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 9
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 claims description 9
- 238000000746 purification Methods 0.000 claims description 9
- 230000001225 therapeutic effect Effects 0.000 claims description 9
- 108010047303 von Willebrand Factor Proteins 0.000 claims description 9
- 102100036537 von Willebrand factor Human genes 0.000 claims description 9
- 229960001134 von willebrand factor Drugs 0.000 claims description 9
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 claims description 8
- 241000124008 Mammalia Species 0.000 claims description 8
- 210000001744 T-lymphocyte Anatomy 0.000 claims description 8
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 claims description 8
- 239000004473 Threonine Substances 0.000 claims description 8
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 claims description 8
- 229960000310 isoleucine Drugs 0.000 claims description 8
- 230000002265 prevention Effects 0.000 claims description 8
- VAIWPXWHWAPYDF-FXQIFTODSA-N Glu-Asp-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O VAIWPXWHWAPYDF-FXQIFTODSA-N 0.000 claims description 7
- MQGGXGKQSVEQHR-KKUMJFAQSA-N Tyr-Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 MQGGXGKQSVEQHR-KKUMJFAQSA-N 0.000 claims description 7
- 239000012530 fluid Substances 0.000 claims description 7
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 claims description 6
- DTLLNDVORUEOTM-WDCWCFNPSA-N Glu-Thr-Lys Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(O)=O DTLLNDVORUEOTM-WDCWCFNPSA-N 0.000 claims description 6
- 108010021625 Immunoglobulin Fragments Proteins 0.000 claims description 6
- 102000008394 Immunoglobulin Fragments Human genes 0.000 claims description 6
- NFDYGNFETJVMSE-BQBZGAKWSA-N Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](N)CO NFDYGNFETJVMSE-BQBZGAKWSA-N 0.000 claims description 6
- 235000009582 asparagine Nutrition 0.000 claims description 6
- 229960001230 asparagine Drugs 0.000 claims description 6
- 108010047857 aspartylglycine Proteins 0.000 claims description 6
- 108010068265 aspartyltyrosine Proteins 0.000 claims description 6
- 210000003719 b-lymphocyte Anatomy 0.000 claims description 6
- 108010078144 glutaminyl-glycine Proteins 0.000 claims description 6
- 108010015792 glycyllysine Proteins 0.000 claims description 6
- 108010004914 prolylarginine Proteins 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 108010061238 threonyl-glycine Proteins 0.000 claims description 6
- 239000003937 drug carrier Substances 0.000 claims description 5
- 239000012528 membrane Substances 0.000 claims description 5
- 102000014914 Carrier Proteins Human genes 0.000 claims description 4
- 108010078791 Carrier Proteins Proteins 0.000 claims description 4
- JHSRGEODDALISP-XVSYOHENSA-N Phe-Thr-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O JHSRGEODDALISP-XVSYOHENSA-N 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- NKJBKNVQHBZUIX-ACZMJKKPSA-N Ala-Gln-Asp Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O NKJBKNVQHBZUIX-ACZMJKKPSA-N 0.000 claims description 3
- 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 claims description 3
- VBRDBGCROKWTPV-XHNCKOQMSA-N Ala-Glu-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N VBRDBGCROKWTPV-XHNCKOQMSA-N 0.000 claims description 3
- HQJKCXHQNUCKMY-GHCJXIJMSA-N Ala-Ile-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](C)N HQJKCXHQNUCKMY-GHCJXIJMSA-N 0.000 claims description 3
- CKLDHDOIYBVUNP-KBIXCLLPSA-N Ala-Ile-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O CKLDHDOIYBVUNP-KBIXCLLPSA-N 0.000 claims description 3
- MSWSRLGNLKHDEI-ACZMJKKPSA-N Ala-Ser-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O MSWSRLGNLKHDEI-ACZMJKKPSA-N 0.000 claims description 3
- JSHVMZANPXCDTL-GMOBBJLQSA-N Arg-Asp-Ile Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JSHVMZANPXCDTL-GMOBBJLQSA-N 0.000 claims description 3
- JUWQNWXEGDYCIE-YUMQZZPRSA-N Arg-Gln-Gly Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O JUWQNWXEGDYCIE-YUMQZZPRSA-N 0.000 claims description 3
- OBFTYSPXDRROQO-SRVKXCTJSA-N Arg-Gln-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CCCN=C(N)N OBFTYSPXDRROQO-SRVKXCTJSA-N 0.000 claims description 3
- UAOSDDXCTBIPCA-QXEWZRGKSA-N Arg-Ile-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CCCN=C(N)N)N UAOSDDXCTBIPCA-QXEWZRGKSA-N 0.000 claims description 3
- SSZGOKWBHLOCHK-DCAQKATOSA-N Arg-Lys-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCCN=C(N)N SSZGOKWBHLOCHK-DCAQKATOSA-N 0.000 claims description 3
- NYDIVDKTULRINZ-AVGNSLFASA-N Arg-Met-Lys Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N NYDIVDKTULRINZ-AVGNSLFASA-N 0.000 claims description 3
- GXMSVVBIAMWMKO-BQBZGAKWSA-N Asn-Arg-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CCCN=C(N)N GXMSVVBIAMWMKO-BQBZGAKWSA-N 0.000 claims description 3
- LJUOLNXOWSWGKF-ACZMJKKPSA-N Asn-Asn-Glu Chemical compound C(CC(=O)O)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)N)N LJUOLNXOWSWGKF-ACZMJKKPSA-N 0.000 claims description 3
- WQSCVMQDZYTFQU-FXQIFTODSA-N Asn-Cys-Arg Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O WQSCVMQDZYTFQU-FXQIFTODSA-N 0.000 claims description 3
- FAEFJTCTNZTPHX-ACZMJKKPSA-N Asn-Gln-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O FAEFJTCTNZTPHX-ACZMJKKPSA-N 0.000 claims description 3
- QIRJQYQOIKBPBZ-IHRRRGAJSA-N Asn-Tyr-Arg 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](CCCNC(N)=N)C(O)=O QIRJQYQOIKBPBZ-IHRRRGAJSA-N 0.000 claims description 3
- JZLFYAAGGYMRIK-BYULHYEWSA-N Asn-Val-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O JZLFYAAGGYMRIK-BYULHYEWSA-N 0.000 claims description 3
- KRXIWXCXOARFNT-ZLUOBGJFSA-N Asp-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC(O)=O KRXIWXCXOARFNT-ZLUOBGJFSA-N 0.000 claims description 3
- JGDBHIVECJGXJA-FXQIFTODSA-N Asp-Asp-Arg Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O JGDBHIVECJGXJA-FXQIFTODSA-N 0.000 claims description 3
- SBHUBSDEZQFJHJ-CIUDSAMLSA-N Asp-Asp-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC(O)=O SBHUBSDEZQFJHJ-CIUDSAMLSA-N 0.000 claims description 3
- XAJRHVUUVUPFQL-ACZMJKKPSA-N Asp-Glu-Asp Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O XAJRHVUUVUPFQL-ACZMJKKPSA-N 0.000 claims description 3
- VFUXXFVCYZPOQG-WDSKDSINSA-N Asp-Glu-Gly Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O VFUXXFVCYZPOQG-WDSKDSINSA-N 0.000 claims description 3
- PZXPWHFYZXTFBI-YUMQZZPRSA-N Asp-Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC(O)=O PZXPWHFYZXTFBI-YUMQZZPRSA-N 0.000 claims description 3
- MYLZFUMPZCPJCJ-NHCYSSNCSA-N Asp-Lys-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O MYLZFUMPZCPJCJ-NHCYSSNCSA-N 0.000 claims description 3
- LIJXJYGRSRWLCJ-IHRRRGAJSA-N Asp-Phe-Arg Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O LIJXJYGRSRWLCJ-IHRRRGAJSA-N 0.000 claims description 3
- RVMXMLSYBTXCAV-VEVYYDQMSA-N Asp-Pro-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O RVMXMLSYBTXCAV-VEVYYDQMSA-N 0.000 claims description 3
- DWBZEJHQQIURML-IMJSIDKUSA-N Asp-Ser Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(O)=O DWBZEJHQQIURML-IMJSIDKUSA-N 0.000 claims description 3
- FIAKNCXQFFKSSI-ZLUOBGJFSA-N Asp-Ser-Cys Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(O)=O FIAKNCXQFFKSSI-ZLUOBGJFSA-N 0.000 claims description 3
- 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 claims description 3
- 102100022641 Coagulation factor IX Human genes 0.000 claims description 3
- SZQCDCKIGWQAQN-FXQIFTODSA-N Cys-Arg-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O SZQCDCKIGWQAQN-FXQIFTODSA-N 0.000 claims description 3
- UCMIKRLLIOVDRJ-XKBZYTNZSA-N Cys-Gln-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CS)N)O UCMIKRLLIOVDRJ-XKBZYTNZSA-N 0.000 claims description 3
- 238000009007 Diagnostic Kit Methods 0.000 claims description 3
- 108010076282 Factor IX Proteins 0.000 claims description 3
- 108010014173 Factor X Proteins 0.000 claims description 3
- LZRMPXRYLLTAJX-GUBZILKMSA-N Gln-Arg-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O LZRMPXRYLLTAJX-GUBZILKMSA-N 0.000 claims description 3
- ZPDVKYLJTOFQJV-WDSKDSINSA-N Gln-Asn-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O ZPDVKYLJTOFQJV-WDSKDSINSA-N 0.000 claims description 3
- BTSPOOHJBYJRKO-CIUDSAMLSA-N Gln-Asp-Arg Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O BTSPOOHJBYJRKO-CIUDSAMLSA-N 0.000 claims description 3
- VSXBYIJUAXPAAL-WDSKDSINSA-N Gln-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H](N)CCC(N)=O VSXBYIJUAXPAAL-WDSKDSINSA-N 0.000 claims description 3
- IKFZXRLDMYWNBU-YUMQZZPRSA-N Gln-Gly-Arg Chemical compound NC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCN=C(N)N IKFZXRLDMYWNBU-YUMQZZPRSA-N 0.000 claims description 3
- CELXWPDNIGWCJN-WDCWCFNPSA-N Gln-Lys-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O CELXWPDNIGWCJN-WDCWCFNPSA-N 0.000 claims description 3
- XQDGOJPVMSWZSO-SRVKXCTJSA-N Gln-Pro-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CCC(=O)N)N XQDGOJPVMSWZSO-SRVKXCTJSA-N 0.000 claims description 3
- ARYKRXHBIPLULY-XKBZYTNZSA-N Gln-Thr-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O ARYKRXHBIPLULY-XKBZYTNZSA-N 0.000 claims description 3
- LKDIBBOKUAASNP-FXQIFTODSA-N Glu-Ala-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O LKDIBBOKUAASNP-FXQIFTODSA-N 0.000 claims description 3
- MLCPTRRNICEKIS-FXQIFTODSA-N Glu-Asn-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O MLCPTRRNICEKIS-FXQIFTODSA-N 0.000 claims description 3
- AUTNXSQEVVHSJK-YVNDNENWSA-N Glu-Glu-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CCC(O)=O AUTNXSQEVVHSJK-YVNDNENWSA-N 0.000 claims description 3
- LHIPZASLKPYDPI-AVGNSLFASA-N Glu-Phe-Asp Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(O)=O LHIPZASLKPYDPI-AVGNSLFASA-N 0.000 claims description 3
- 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 claims description 3
- MFVQGXGQRIXBPK-WDSKDSINSA-N Gly-Ala-Glu Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O MFVQGXGQRIXBPK-WDSKDSINSA-N 0.000 claims description 3
- RJIVPOXLQFJRTG-LURJTMIESA-N Gly-Arg-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CCCN=C(N)N RJIVPOXLQFJRTG-LURJTMIESA-N 0.000 claims description 3
- WJZLEENECIOOSA-WDSKDSINSA-N Gly-Asn-Gln Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)O WJZLEENECIOOSA-WDSKDSINSA-N 0.000 claims description 3
- XCLCVBYNGXEVDU-WHFBIAKZSA-N Gly-Asn-Ser Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O XCLCVBYNGXEVDU-WHFBIAKZSA-N 0.000 claims description 3
- LCNXZQROPKFGQK-WHFBIAKZSA-N Gly-Asp-Ser Chemical compound NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O LCNXZQROPKFGQK-WHFBIAKZSA-N 0.000 claims description 3
- QGZSAHIZRQHCEQ-QWRGUYRKSA-N Gly-Asp-Tyr Chemical compound NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 QGZSAHIZRQHCEQ-QWRGUYRKSA-N 0.000 claims description 3
- XXGQRGQPGFYECI-WDSKDSINSA-N Gly-Cys-Glu Chemical compound NCC(=O)N[C@@H](CS)C(=O)N[C@H](C(O)=O)CCC(O)=O XXGQRGQPGFYECI-WDSKDSINSA-N 0.000 claims description 3
- ZKLYPEGLWFVRGF-IUCAKERBSA-N Gly-His-Gln Chemical compound [H]NCC(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(N)=O)C(O)=O ZKLYPEGLWFVRGF-IUCAKERBSA-N 0.000 claims description 3
- HJARVELKOSZUEW-YUMQZZPRSA-N Gly-Pro-Gln Chemical compound [H]NCC(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(O)=O HJARVELKOSZUEW-YUMQZZPRSA-N 0.000 claims description 3
- GLACUWHUYFBSPJ-FJXKBIBVSA-N Gly-Pro-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)CN GLACUWHUYFBSPJ-FJXKBIBVSA-N 0.000 claims description 3
- FNXSYBOHALPRHV-ONGXEEELSA-N Gly-Val-Lys Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN FNXSYBOHALPRHV-ONGXEEELSA-N 0.000 claims description 3
- AFMOTCMSEBITOE-YEPSODPASA-N Gly-Val-Thr Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O AFMOTCMSEBITOE-YEPSODPASA-N 0.000 claims description 3
- HRGGKHFHRSFSDE-CIUDSAMLSA-N His-Asn-Ser Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CO)C(=O)O)N HRGGKHFHRSFSDE-CIUDSAMLSA-N 0.000 claims description 3
- OWYIDJCNRWRSJY-QTKMDUPCSA-N His-Pro-Thr Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O OWYIDJCNRWRSJY-QTKMDUPCSA-N 0.000 claims description 3
- JMSONHOUHFDOJH-GUBZILKMSA-N His-Ser-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CN=CN1 JMSONHOUHFDOJH-GUBZILKMSA-N 0.000 claims description 3
- JATYGDHMDRAISQ-KKUMJFAQSA-N His-Tyr-Ser Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(O)=O JATYGDHMDRAISQ-KKUMJFAQSA-N 0.000 claims description 3
- NULSANWBUWLTKN-NAKRPEOUSA-N Ile-Arg-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(=O)O)N NULSANWBUWLTKN-NAKRPEOUSA-N 0.000 claims description 3
- UASTVUQJMLZWGG-PEXQALLHSA-N Ile-His-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)NCC(=O)O)N UASTVUQJMLZWGG-PEXQALLHSA-N 0.000 claims description 3
- XDUVMJCBYUKNFJ-MXAVVETBSA-N Ile-Lys-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N XDUVMJCBYUKNFJ-MXAVVETBSA-N 0.000 claims description 3
- CKRFDMPBSWYOBT-PPCPHDFISA-N Ile-Lys-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N CKRFDMPBSWYOBT-PPCPHDFISA-N 0.000 claims description 3
- JZNVOBUNTWNZPW-GHCJXIJMSA-N Ile-Ser-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)O)C(=O)O)N JZNVOBUNTWNZPW-GHCJXIJMSA-N 0.000 claims description 3
- PXKACEXYLPBMAD-JBDRJPRFSA-N Ile-Ser-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)O)N PXKACEXYLPBMAD-JBDRJPRFSA-N 0.000 claims description 3
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 claims description 3
- DBVWMYGBVFCRBE-CIUDSAMLSA-N Leu-Asn-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O DBVWMYGBVFCRBE-CIUDSAMLSA-N 0.000 claims description 3
- CSFVADKICPDRRF-KKUMJFAQSA-N Leu-His-Leu Chemical compound CC(C)C[C@H]([NH3+])C(=O)N[C@H](C(=O)N[C@@H](CC(C)C)C([O-])=O)CC1=CN=CN1 CSFVADKICPDRRF-KKUMJFAQSA-N 0.000 claims description 3
- RXGLHDWAZQECBI-SRVKXCTJSA-N Leu-Leu-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O RXGLHDWAZQECBI-SRVKXCTJSA-N 0.000 claims description 3
- MJWVXZABPOKJJF-ACRUOGEOSA-N Leu-Phe-Phe Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O MJWVXZABPOKJJF-ACRUOGEOSA-N 0.000 claims description 3
- RVOMPSJXSRPFJT-DCAQKATOSA-N Lys-Ala-Arg Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O RVOMPSJXSRPFJT-DCAQKATOSA-N 0.000 claims description 3
- ABHIXYDMILIUKV-CIUDSAMLSA-N Lys-Asn-Asn Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O ABHIXYDMILIUKV-CIUDSAMLSA-N 0.000 claims description 3
- 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 claims description 3
- LLSUNJYOSCOOEB-GUBZILKMSA-N Lys-Glu-Asp Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O LLSUNJYOSCOOEB-GUBZILKMSA-N 0.000 claims description 3
- PLDJDCJLRCYPJB-VOAKCMCISA-N Lys-Lys-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PLDJDCJLRCYPJB-VOAKCMCISA-N 0.000 claims description 3
- MGKFCQFVPKOWOL-CIUDSAMLSA-N Lys-Ser-Asp Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)O)C(=O)O)N MGKFCQFVPKOWOL-CIUDSAMLSA-N 0.000 claims description 3
- UIJVKVHLCQSPOJ-XIRDDKMYSA-N Lys-Ser-Trp Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(O)=O UIJVKVHLCQSPOJ-XIRDDKMYSA-N 0.000 claims description 3
- OEYKVQKYCHATHO-SZMVWBNQSA-N Lys-Trp-Gln Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CCCCN)N OEYKVQKYCHATHO-SZMVWBNQSA-N 0.000 claims description 3
- NYTDJEZBAAFLLG-IHRRRGAJSA-N Lys-Val-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(O)=O NYTDJEZBAAFLLG-IHRRRGAJSA-N 0.000 claims description 3
- ULNXMMYXQKGNPG-LPEHRKFASA-N Met-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCSC)N ULNXMMYXQKGNPG-LPEHRKFASA-N 0.000 claims description 3
- WXHHTBVYQOSYSL-FXQIFTODSA-N Met-Ala-Ser Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O WXHHTBVYQOSYSL-FXQIFTODSA-N 0.000 claims description 3
- KQBJYJXPZBNEIK-DCAQKATOSA-N Met-Glu-Arg Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCNC(N)=N KQBJYJXPZBNEIK-DCAQKATOSA-N 0.000 claims description 3
- WTHGNAAQXISJHP-AVGNSLFASA-N Met-Lys-Val Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O WTHGNAAQXISJHP-AVGNSLFASA-N 0.000 claims description 3
- 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 claims description 3
- 108010079364 N-glycylalanine Proteins 0.000 claims description 3
- WLYPRKLMRIYGPP-JYJNAYRXSA-N Phe-Lys-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC1=CC=CC=C1 WLYPRKLMRIYGPP-JYJNAYRXSA-N 0.000 claims description 3
- 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 claims description 3
- RGMLUHANLDVMPB-ULQDDVLXSA-N Phe-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N RGMLUHANLDVMPB-ULQDDVLXSA-N 0.000 claims description 3
- CYQQWUPHIZVCNY-GUBZILKMSA-N Pro-Arg-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O CYQQWUPHIZVCNY-GUBZILKMSA-N 0.000 claims description 3
- OBVCYFIHIIYIQF-CIUDSAMLSA-N Pro-Asn-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O OBVCYFIHIIYIQF-CIUDSAMLSA-N 0.000 claims description 3
- HJSCRFZVGXAGNG-SRVKXCTJSA-N Pro-Gln-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H]1CCCN1 HJSCRFZVGXAGNG-SRVKXCTJSA-N 0.000 claims description 3
- FRKBNXCFJBPJOL-GUBZILKMSA-N Pro-Glu-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O FRKBNXCFJBPJOL-GUBZILKMSA-N 0.000 claims description 3
- FXGIMYRVJJEIIM-UWVGGRQHSA-N Pro-Leu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CCCN1 FXGIMYRVJJEIIM-UWVGGRQHSA-N 0.000 claims description 3
- BGWKULMLUIUPKY-BQBZGAKWSA-N Pro-Ser-Gly Chemical compound OC(=O)CNC(=O)[C@H](CO)NC(=O)[C@@H]1CCCN1 BGWKULMLUIUPKY-BQBZGAKWSA-N 0.000 claims description 3
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims description 3
- NRCJWSGXMAPYQX-LPEHRKFASA-N Ser-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CO)N)C(=O)O NRCJWSGXMAPYQX-LPEHRKFASA-N 0.000 claims description 3
- XVAUJOAYHWWNQF-ZLUOBGJFSA-N Ser-Asn-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(O)=O XVAUJOAYHWWNQF-ZLUOBGJFSA-N 0.000 claims description 3
- OOKCGAYXSNJBGQ-ZLUOBGJFSA-N Ser-Asn-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O OOKCGAYXSNJBGQ-ZLUOBGJFSA-N 0.000 claims description 3
- VGNYHOBZJKWRGI-CIUDSAMLSA-N Ser-Asn-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CO VGNYHOBZJKWRGI-CIUDSAMLSA-N 0.000 claims description 3
- MESDJCNHLZBMEP-ZLUOBGJFSA-N Ser-Asp-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O MESDJCNHLZBMEP-ZLUOBGJFSA-N 0.000 claims description 3
- VAIZFHMTBFYJIA-ACZMJKKPSA-N Ser-Asp-Gln Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCC(N)=O VAIZFHMTBFYJIA-ACZMJKKPSA-N 0.000 claims description 3
- KNCJWSPMTFFJII-ZLUOBGJFSA-N Ser-Cys-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(O)=O)C(O)=O KNCJWSPMTFFJII-ZLUOBGJFSA-N 0.000 claims 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 claims description 3
- VDVYTKZBMFADQH-AVGNSLFASA-N Ser-Gln-Tyr Chemical compound OC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 VDVYTKZBMFADQH-AVGNSLFASA-N 0.000 claims description 3
- XXNYYSXNXCJYKX-DCAQKATOSA-N Ser-Leu-Met Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(O)=O XXNYYSXNXCJYKX-DCAQKATOSA-N 0.000 claims description 3
- GZSZPKSBVAOGIE-CIUDSAMLSA-N Ser-Lys-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O GZSZPKSBVAOGIE-CIUDSAMLSA-N 0.000 claims description 3
- PPQRSMGDOHLTBE-UWVGGRQHSA-N Ser-Phe Chemical compound OC[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 PPQRSMGDOHLTBE-UWVGGRQHSA-N 0.000 claims description 3
- FZEUTKVQGMVGHW-AVGNSLFASA-N Ser-Phe-Gln Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O FZEUTKVQGMVGHW-AVGNSLFASA-N 0.000 claims description 3
- FBLNYDYPCLFTSP-IXOXFDKPSA-N Ser-Phe-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O FBLNYDYPCLFTSP-IXOXFDKPSA-N 0.000 claims description 3
- NUEHQDHDLDXCRU-GUBZILKMSA-N Ser-Pro-Arg Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(O)=O NUEHQDHDLDXCRU-GUBZILKMSA-N 0.000 claims description 3
- AZWNCEBQZXELEZ-FXQIFTODSA-N Ser-Pro-Ser Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O AZWNCEBQZXELEZ-FXQIFTODSA-N 0.000 claims description 3
- SRSPTFBENMJHMR-WHFBIAKZSA-N Ser-Ser-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SRSPTFBENMJHMR-WHFBIAKZSA-N 0.000 claims description 3
- VAIWUNAAPZZGRI-IHPCNDPISA-N Ser-Trp-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)NC(=O)[C@H](CO)N VAIWUNAAPZZGRI-IHPCNDPISA-N 0.000 claims description 3
- RTXKJFWHEBTABY-IHPCNDPISA-N Ser-Trp-Tyr Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC3=CC=C(C=C3)O)C(=O)O)NC(=O)[C@H](CO)N RTXKJFWHEBTABY-IHPCNDPISA-N 0.000 claims description 3
- HKHCTNFKZXAMIF-KKUMJFAQSA-N Ser-Tyr-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)CC1=CC=C(O)C=C1 HKHCTNFKZXAMIF-KKUMJFAQSA-N 0.000 claims description 3
- ZVBCMFDJIMUELU-BZSNNMDCSA-N Ser-Tyr-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CO)N ZVBCMFDJIMUELU-BZSNNMDCSA-N 0.000 claims description 3
- 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 claims description 3
- 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 claims description 3
- 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 claims description 3
- 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 claims description 3
- JEDIEMIJYSRUBB-FOHZUACHSA-N Thr-Asp-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O JEDIEMIJYSRUBB-FOHZUACHSA-N 0.000 claims description 3
- 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 claims description 3
- UHBPFYOQQPFKQR-JHEQGTHGSA-N Thr-Gln-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O UHBPFYOQQPFKQR-JHEQGTHGSA-N 0.000 claims description 3
- CQNFRKAKGDSJFR-NUMRIWBASA-N Thr-Glu-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O CQNFRKAKGDSJFR-NUMRIWBASA-N 0.000 claims description 3
- JKGGPMOUIAAJAA-YEPSODPASA-N Thr-Gly-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O JKGGPMOUIAAJAA-YEPSODPASA-N 0.000 claims description 3
- BQBCIBCLXBKYHW-CSMHCCOUSA-N Thr-Leu Chemical compound CC(C)C[C@@H](C([O-])=O)NC(=O)[C@@H]([NH3+])[C@@H](C)O BQBCIBCLXBKYHW-CSMHCCOUSA-N 0.000 claims description 3
- AMXMBCAXAZUCFA-RHYQMDGZSA-N Thr-Leu-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O AMXMBCAXAZUCFA-RHYQMDGZSA-N 0.000 claims description 3
- HOVLHEKTGVIKAP-WDCWCFNPSA-N Thr-Leu-Gln Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O HOVLHEKTGVIKAP-WDCWCFNPSA-N 0.000 claims description 3
- BDGBHYCAZJPLHX-HJGDQZAQSA-N Thr-Lys-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O BDGBHYCAZJPLHX-HJGDQZAQSA-N 0.000 claims description 3
- ZSPQUTWLWGWTPS-HJGDQZAQSA-N Thr-Lys-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(O)=O ZSPQUTWLWGWTPS-HJGDQZAQSA-N 0.000 claims description 3
- JWQNAFHCXKVZKZ-UVOCVTCTSA-N Thr-Lys-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O JWQNAFHCXKVZKZ-UVOCVTCTSA-N 0.000 claims description 3
- MFMGPEKYBXFIRF-SUSMZKCASA-N Thr-Thr-Gln Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(O)=O MFMGPEKYBXFIRF-SUSMZKCASA-N 0.000 claims description 3
- BEZTUFWTPVOROW-KJEVXHAQSA-N Thr-Tyr-Arg Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N)O BEZTUFWTPVOROW-KJEVXHAQSA-N 0.000 claims description 3
- VIWQOOBRKCGSDK-RYQLBKOJSA-N Trp-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N)C(=O)O VIWQOOBRKCGSDK-RYQLBKOJSA-N 0.000 claims description 3
- BEIGSKUPTIFYRZ-SRVKXCTJSA-N Tyr-Asp-Asp Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N)O BEIGSKUPTIFYRZ-SRVKXCTJSA-N 0.000 claims description 3
- 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 claims description 3
- JLKVWTICWVWGSK-JYJNAYRXSA-N Tyr-Lys-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 JLKVWTICWVWGSK-JYJNAYRXSA-N 0.000 claims description 3
- HRHYJNLMIJWGLF-BZSNNMDCSA-N Tyr-Ser-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=C(O)C=C1 HRHYJNLMIJWGLF-BZSNNMDCSA-N 0.000 claims description 3
- GBESYURLQOYWLU-LAEOZQHASA-N Val-Glu-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N GBESYURLQOYWLU-LAEOZQHASA-N 0.000 claims description 3
- JAKHAONCJJZVHT-DCAQKATOSA-N Val-Lys-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)O)N JAKHAONCJJZVHT-DCAQKATOSA-N 0.000 claims description 3
- UZFNHAXYMICTBU-DZKIICNBSA-N Val-Phe-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N UZFNHAXYMICTBU-DZKIICNBSA-N 0.000 claims description 3
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 claims description 3
- 108010040443 aspartyl-aspartic acid Proteins 0.000 claims description 3
- 108010093581 aspartyl-proline Proteins 0.000 claims description 3
- 108010038633 aspartylglutamate Proteins 0.000 claims description 3
- 108010092854 aspartyllysine Proteins 0.000 claims description 3
- 108010060199 cysteinylproline Proteins 0.000 claims description 3
- 229960004222 factor ix Drugs 0.000 claims description 3
- 108010042598 glutamyl-aspartyl-glycine Proteins 0.000 claims description 3
- 108010055341 glutamyl-glutamic acid Proteins 0.000 claims description 3
- 108010049041 glutamylalanine Proteins 0.000 claims description 3
- 108010079547 glutamylmethionine Proteins 0.000 claims description 3
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 claims description 3
- 108010090037 glycyl-alanyl-isoleucine Proteins 0.000 claims description 3
- 108010020688 glycylhistidine Proteins 0.000 claims description 3
- 108010050848 glycylleucine Proteins 0.000 claims description 3
- 108010044348 lysyl-glutamyl-aspartic acid Proteins 0.000 claims description 3
- 108010054155 lysyllysine Proteins 0.000 claims description 3
- 108010038320 lysylphenylalanine Proteins 0.000 claims description 3
- 108010016686 methionyl-alanyl-serine Proteins 0.000 claims description 3
- 108700042769 prolyl-leucyl-glycine Proteins 0.000 claims description 3
- 108010029384 tryptophyl-histidine Proteins 0.000 claims description 3
- QBQVKUNBCAFXSV-ULQDDVLXSA-N Arg-Lys-Tyr Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 QBQVKUNBCAFXSV-ULQDDVLXSA-N 0.000 claims description 2
- NHMRJKKAVMENKJ-WDCWCFNPSA-N Gln-Thr-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O NHMRJKKAVMENKJ-WDCWCFNPSA-N 0.000 claims description 2
- RAUDKMVXNOWDLS-WDSKDSINSA-N Glu-Gly-Ser Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O RAUDKMVXNOWDLS-WDSKDSINSA-N 0.000 claims description 2
- 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 claims description 2
- KWTVLKBOQATPHJ-SRVKXCTJSA-N Leu-Ala-Lys Chemical compound C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(C)C)N KWTVLKBOQATPHJ-SRVKXCTJSA-N 0.000 claims description 2
- FIXILCYTSAUERA-FXQIFTODSA-N Ser-Ala-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FIXILCYTSAUERA-FXQIFTODSA-N 0.000 claims description 2
- NHUHCSRWZMLRLA-UHFFFAOYSA-N Sulfisoxazole Chemical compound CC1=NOC(NS(=O)(=O)C=2C=CC(N)=CC=2)=C1C NHUHCSRWZMLRLA-UHFFFAOYSA-N 0.000 claims description 2
- 108010008685 alanyl-glutamyl-aspartic acid Proteins 0.000 claims description 2
- 108010009298 lysylglutamic acid Proteins 0.000 claims description 2
- 108010051110 tyrosyl-lysine Proteins 0.000 claims description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims 1
- MKJBPDLENBUHQU-CIUDSAMLSA-N Asn-Ser-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O MKJBPDLENBUHQU-CIUDSAMLSA-N 0.000 claims 1
- BKOIIURTQAJHAT-GUBZILKMSA-N Asp-Pro-Pro Chemical compound OC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 BKOIIURTQAJHAT-GUBZILKMSA-N 0.000 claims 1
- IEWBEPKLKUXQBU-VOAKCMCISA-N Leu-Leu-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O IEWBEPKLKUXQBU-VOAKCMCISA-N 0.000 claims 1
- FHJQROWZEJFZPO-SRVKXCTJSA-N Pro-Val-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 FHJQROWZEJFZPO-SRVKXCTJSA-N 0.000 claims 1
- 108010090894 prolylleucine Proteins 0.000 claims 1
- 102100026735 Coagulation factor VIII Human genes 0.000 description 115
- 101000911390 Homo sapiens Coagulation factor VIII Proteins 0.000 description 115
- 241000283973 Oryctolagus cuniculus Species 0.000 description 27
- 230000000694 effects Effects 0.000 description 26
- 210000002381 plasma Anatomy 0.000 description 20
- 238000002360 preparation method Methods 0.000 description 20
- 229960004072 thrombin Drugs 0.000 description 16
- 108090000190 Thrombin Proteins 0.000 description 15
- 108090000623 proteins and genes Proteins 0.000 description 14
- 108060003951 Immunoglobulin Proteins 0.000 description 12
- 229920002684 Sepharose Polymers 0.000 description 12
- 102000018358 immunoglobulin Human genes 0.000 description 12
- 235000018102 proteins Nutrition 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 12
- 238000002965 ELISA Methods 0.000 description 10
- 230000002788 anti-peptide Effects 0.000 description 10
- 238000003556 assay Methods 0.000 description 10
- 230000002401 inhibitory effect Effects 0.000 description 10
- 229940072221 immunoglobulins Drugs 0.000 description 9
- 230000005764 inhibitory process Effects 0.000 description 9
- 230000015572 biosynthetic process Effects 0.000 description 8
- 238000003119 immunoblot Methods 0.000 description 8
- 208000009292 Hemophilia A Diseases 0.000 description 7
- 238000012512 characterization method Methods 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 7
- 239000012141 concentrate Substances 0.000 description 7
- 230000002163 immunogen Effects 0.000 description 7
- 230000009257 reactivity Effects 0.000 description 7
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 6
- 102100022851 Rab5 GDP/GTP exchange factor Human genes 0.000 description 6
- 101710203837 Replication-associated protein Proteins 0.000 description 6
- 230000004913 activation Effects 0.000 description 6
- 230000035602 clotting Effects 0.000 description 6
- 230000017854 proteolysis Effects 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- 102000002110 C2 domains Human genes 0.000 description 5
- 108050009459 C2 domains Proteins 0.000 description 5
- 206010053567 Coagulopathies Diseases 0.000 description 5
- 108010032597 Cohn fraction II Proteins 0.000 description 5
- 125000000539 amino acid group Chemical group 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 108010045069 keyhole-limpet hemocyanin Proteins 0.000 description 5
- 229940092253 ovalbumin Drugs 0.000 description 5
- 108020003175 receptors Proteins 0.000 description 5
- 238000011084 recovery Methods 0.000 description 5
- 238000002560 therapeutic procedure Methods 0.000 description 5
- 238000001262 western blot Methods 0.000 description 5
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- 201000003542 Factor VIII deficiency Diseases 0.000 description 4
- 101001130441 Homo sapiens Ras-related protein Rap-2a Proteins 0.000 description 4
- 108010058846 Ovalbumin Proteins 0.000 description 4
- 102100031420 Ras-related protein Rap-2a Human genes 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 238000001042 affinity chromatography Methods 0.000 description 4
- 229940098773 bovine serum albumin Drugs 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 230000028993 immune response Effects 0.000 description 4
- 230000005847 immunogenicity Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 229940126619 mouse monoclonal antibody Drugs 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 102100040197 Apolipoprotein A-V Human genes 0.000 description 3
- 102100037373 DNA-(apurinic or apyrimidinic site) endonuclease Human genes 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 229940124135 Factor VIII inhibitor Drugs 0.000 description 3
- 101710088570 Flagellar hook-associated protein 1 Proteins 0.000 description 3
- 208000031220 Hemophilia Diseases 0.000 description 3
- 101000889974 Homo sapiens Apolipoprotein A-V Proteins 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 239000002671 adjuvant Substances 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000000427 antigen Substances 0.000 description 3
- 108091007433 antigens Proteins 0.000 description 3
- 102000036639 antigens Human genes 0.000 description 3
- 229960002685 biotin Drugs 0.000 description 3
- 235000020958 biotin Nutrition 0.000 description 3
- 239000011616 biotin Substances 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 238000010353 genetic engineering Methods 0.000 description 3
- 230000036046 immunoreaction Effects 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 229940008228 intravenous immunoglobulins Drugs 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000002823 phage display Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 239000007790 solid phase Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-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
- 101100382835 Arabidopsis thaliana CCC1 gene Proteins 0.000 description 2
- 101100036901 Arabidopsis thaliana RPL40B gene Proteins 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- 102100031692 GTPase-activating protein and VPS9 domain-containing protein 1 Human genes 0.000 description 2
- 101150009243 HAP1 gene Proteins 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 101100282121 Homo sapiens GAPVD1 gene Proteins 0.000 description 2
- 241001494479 Pecora Species 0.000 description 2
- 238000011579 SCID mouse model Methods 0.000 description 2
- 101100123443 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) HAP4 gene Proteins 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- 239000007984 Tris EDTA buffer Substances 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 238000003491 array Methods 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000005018 casein Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000015271 coagulation Effects 0.000 description 2
- 238000005345 coagulation Methods 0.000 description 2
- 239000000562 conjugate Substances 0.000 description 2
- 230000021615 conjugation Effects 0.000 description 2
- 239000000356 contaminant Substances 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 238000000502 dialysis Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 230000001747 exhibiting effect Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 101150020171 hap5 gene Proteins 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 230000006058 immune tolerance Effects 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- 230000003472 neutralizing effect Effects 0.000 description 2
- 238000012552 review Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- DWHCYDWXLJOFFO-UHFFFAOYSA-N 4-(5-phenylthiophen-2-yl)aniline Chemical compound C1=CC(N)=CC=C1C1=CC=C(C=2C=CC=CC=2)S1 DWHCYDWXLJOFFO-UHFFFAOYSA-N 0.000 description 1
- 101800000112 Acidic peptide Proteins 0.000 description 1
- 108010032595 Antibody Binding Sites Proteins 0.000 description 1
- 101100435006 Arabidopsis thaliana AP1M2 gene Proteins 0.000 description 1
- 101100473191 Arabidopsis thaliana RPN2 gene Proteins 0.000 description 1
- 101100465815 Arabidopsis thaliana RPN3A gene Proteins 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- 108010029697 CD40 Ligand Proteins 0.000 description 1
- 102100032937 CD40 ligand Human genes 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 108010075016 Ceruloplasmin Proteins 0.000 description 1
- 102100023321 Ceruloplasmin Human genes 0.000 description 1
- 108010014172 Factor V Proteins 0.000 description 1
- 108010074860 Factor Xa Proteins 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 101710088566 Flagellar hook-associated protein 2 Proteins 0.000 description 1
- 101710088564 Flagellar hook-associated protein 3 Proteins 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
- 208000034826 Genetic Predisposition to Disease Diseases 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 102100039384 Huntingtin-associated protein 1 Human genes 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- 108090001090 Lectins Proteins 0.000 description 1
- 102000004856 Lectins Human genes 0.000 description 1
- 108091054437 MHC class I family Proteins 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 102100022201 Nuclear transcription factor Y subunit beta Human genes 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 108010067902 Peptide Library Proteins 0.000 description 1
- 102000029797 Prion Human genes 0.000 description 1
- 108091000054 Prion Proteins 0.000 description 1
- 101800004937 Protein C Proteins 0.000 description 1
- 102000017975 Protein C Human genes 0.000 description 1
- 101800001700 Saposin-D Proteins 0.000 description 1
- 208000026552 Severe hemophilia A Diseases 0.000 description 1
- 108010088160 Staphylococcal Protein A Proteins 0.000 description 1
- 230000024932 T cell mediated immunity Effects 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- ZOBLBMGJKVJVEV-BZSNNMDCSA-N Tyr-Lys-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)O)N)O ZOBLBMGJKVJVEV-BZSNNMDCSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000001261 affinity purification Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000005875 antibody response Effects 0.000 description 1
- 210000000612 antigen-presenting cell Anatomy 0.000 description 1
- 108010062796 arginyllysine Proteins 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 239000003114 blood coagulation factor Substances 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000006167 equilibration buffer Substances 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 238000011577 humanized mouse model Methods 0.000 description 1
- 230000028996 humoral immune response Effects 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 238000001114 immunoprecipitation Methods 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 239000002523 lectin Substances 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 108091005763 multidomain proteins Proteins 0.000 description 1
- 238000004848 nephelometry Methods 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 229940043504 normal human immunoglobulins Drugs 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 239000000863 peptide conjugate Substances 0.000 description 1
- 239000008024 pharmaceutical diluent Substances 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 230000036515 potency Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000002947 procoagulating effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000002731 protein assay Methods 0.000 description 1
- 229960000856 protein c Drugs 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- 108010012557 prothrombin complex concentrates Proteins 0.000 description 1
- 238000011555 rabbit model Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000009256 replacement therapy Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000011012 sanitization Methods 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 230000009291 secondary effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 238000011830 transgenic mouse model Methods 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 150000003668 tyrosines Chemical class 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6878—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids in eptitope analysis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0005—Vertebrate antigens
- A61K39/0008—Antigens related to auto-immune diseases; Preparations to induce self-tolerance
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/745—Blood coagulation or fibrinolysis factors
- C07K14/755—Factors VIII, e.g. factor VIII C (AHF), factor VIII Ag (VWF)
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/745—Assays involving non-enzymic blood coagulation factors
- G01N2333/755—Factors VIII, e.g. factor VIII C [AHF], factor VIII Ag [VWF]
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
- Y10S530/806—Antigenic peptides or proteins
Definitions
- the present invention relates to the antigenic polypeptide sequences of factor VIII, to fragments and epitopes of these sequences and to the major parts of these epitopes, to the inhibitors which are directed against these sequences, its fragments, its epitopes and/or major parts of these epitopes, and to anti-inhibitors which are directed against the said inhibitors.
- the present invention also relates to a pharmaceutical composition and to a diagnostic device comprising at least one of the above mentioned molecules.
- FVIII is a large multi-domain protein of 2,332 amino acids made up of three structural domains, A, B and C which are arranged in the order A1:a1:A2:a2:B:a3:A3:C1:C2.
- the A domains possess more than 40% homology and are also homologous to ceruloplasmin (for recent review, see Pratt (2000) and Saenko (1999)). 30% homology also exists between the A domains of factor V and FVIII.
- the C domain occurs twice and is reported to be able to bind glyco-conjugates and phospholipids having a net negative charge. It exhibits homology with lectins which are able to bind to negatively charged phospholipids.
- the platelet attachment site has been located in this region (C2 domain) (Foster et al., (1990)).
- antigenic determinants consist of fragments 351-365 (A1 domain-heavy chain), 713-740 (A2 domain), 1670-1684 (A3 domain-light chain) (NH 2 end of the light chain) or else 2303-2332 (C2 domain-light chain) (Foster C, (1990)), fragments 701-750, 1663-1689, 330-472, 1694-1782 (EP-0 202 853), 322-740 and 2170-2322.
- the U.S. Pat. No. 5,744,446 describes an hybrid human/animal Factor VIII having a sequence of amino acids selected from the group of the A2 domain fragments 373-540, 373-508, 445-508, 484-508, 404-508, 489-508 and 484-489, with corresponding sequences of porcine or murine Factor VIII, said hybrid being used for the treatment of Factor VIII deficiencies.
- Self proteins or derived peptides may elicit an immune response if presented to CD4 T cells at inflammatory sites by professional antigen presenting cells.
- FVIII domains were recognized: A3 domain was recognized more strongly and frequently and each domain forms several epitopes.
- FVIII FVIII
- PCC activated PCC
- FVIIa FVIIa
- porcine FVIII may be used to achieve haemostasis in patients with antibodies that do not substantially crossreact with porcine FVIII before or during the treatment (Lollar, 2000).
- a potential alternative approach to inhibit the production of inhibitors is blockade of the T cell/B cell collaboration mediated by through receptor ligand binding signal events (Ewenstein et al, 2000). Preliminary clinical trials were performed using a humanized mouse monoclonal antibody to human T cell CD40 ligand (CD 154).
- a profitable strategy for reducing the level of inhibitors has consisted in subjecting patients to an extracorporeal circulation to enable solid-phase absorption of the total IgG.
- the immunoabsorbant could be sepharose-bound staphylococcal protein A or sepharose-bound polyclonal sheep antibodies to total human immunoglobulin (Knobf and Derfler, 1999).
- the foreign proteins protein A, sheep anti-human Ig
- ICH Topic Q5A, Directive 92/79/EC ICH Topic Q5A, Directive 92/79/EC
- IVIG polyvalent intravenous immunoglobulins
- the present invention aims to obtain antigenic polypeptide sequences of factor VIII, fragments and epitopes of these sequences, whose purpose is to improve the diagnosis and/or therapy (including prevention) of immune disorders (in particular those induced by inhibitors of FVIII and inhibitors of FVIII, especially inhibitors of the binding of the von Willebrand factor (vWf), to the FIX and/or to membrane phospholipids (PL)), and which allows a screening between non-inhibitory and inhibitory anti-FVIII allo- or auto-antibodies (allo- or auto-immunoglobulins).
- vWf von Willebrand factor
- PL membrane phospholipids
- Another aim of the invention is to obtain inhibitors which exhibit an immunoaffinity with these antigenic polypeptide sequences, fragments and/or epitopes, as well as to obtain anti-inhibitors, in particular antibodies or (T)cell receptors, which are directed against the above-mentioned said inhibitors and whose purpose is to improve the diagnosis and/or therapy (or prevention) of immune disorders.
- a further aim of the invention is to obtain said molecules at high purity, in industrial level, without contaminants (viruses, prions, . . . ) and according to the GMP practices in the field of therapy and diagnostics (ICH topic QSA, Directive 92/79/EC, etc.).
- An antigenic polypeptide sequence which is the polypeptide sequence of factor VIII.
- An antigenic polypeptide sequence which lacks the following fragments: alanine 322-serine 750, leucine 1655-arginine 1689, lysine 1694-proline 1782 and possibly the fragment aspartic acid 2170-tyrosine 2332.
- An antigenic fragment of the sequence according to paragraph 1 or 2 which is selected from the group consisting of the polypeptide sequences Al, A2, A3 or C of factor VIII.
- epitope arginine 1648 to tyrosine 1664 inclusive defined by the following sequence:
- epitope threonine 1739 to tyrosine 1748 inclusive defined by the following sequence:
- epitope asparagine 1777 to phenylalanine 1785 inclusive defined by the following sequence:
- epitope glutamic acid 1794 to tyrosine 1815 inclusive defined by the following sequence:
- epitope methionine 1823 to aspartic acid 1831 defined by the following sequence:
- epitope glutamic acid 1885 to phenylalanine 1891 inclusive defined by the following sequence:
- epitope glutamic acid 1885 to alanine 1901 inclusive defined by the following sequence:
- epitope aspartic acid 1909 to arginine 1917 inclusive defined by the following sequence:
- epitope glutamic acid 181 to leucine 192 inclusive defined by the following sequence:
- epitope aspartic acid 203 to alanine 227 inclusive defined by the following sequence:
- epitope aspartic acid 327 to methionine 355 inclusive defined by the following sequence:
- epitope aspartic acid 403 to lysine 425 inclusive defined by the following sequence:
- epitope valine 517 to arginine 527 inclusive defined by the following sequence:
- epitope tyrosine 555 to glutamine 565 inclusive defined by the following sequence:
- epitope histidine 693 to glycine 701 inclusive defined by the following sequence:
- epitope serine 710 to aspartic acid 725 inclusive defined by the following sequence:
- epitope leucine 730 to serine 741 inclusive defined by the following sequence:
- epitope serine 817 to serine 830 inclusive defined by the following sequence:
- epitope isoleucine 2081 to serine 2095 inclusive defined by the following sequence
- epitope tyrosine 2105 to glycine 2121 inclusive defined by the following sequence:
- epitope asparagine acid 2128 to asparagine acid 2138 inclusive defined by the following sequence:
- epitope histidine 2152 to arginine 2163 inclusive defined by the following sequence:
- epitope serine 2181 to asparagine acid 2198 inclusive defined by the following sequence:
- epitope serine 2204 to glutamine 2222 inclusive defined by the following sequence:
- epitope glutamine 2235 to leucine 2251 inclusive defined by the following sequence:
- epitope glycine 2242 to leucine 2251 inclusive defined by the following sequence:
- epitope leucine 2273 to serine 2289 inclusive defined by the following sequence:
- epitope proline 2292 to tyrosine 2305 inclusive defined by the following sequence:
- epitope glutamic acid 2322 to tyrosine 2332 inclusive defined by the following sequence:
- a conformational epitope which contains at least two different epitopes according to any one of the preceding paragraphs 8, 10 and 12.
- a complex comprising a carrier protein or a carrier peptide linked to an element which is selected from the group consisting of the fragment and/or the epitope according to any one of the paragraphs 6 to 14.
- a pharmaceutical composition which comprises an adequate pharmaceutical carrier and at least one element selected from the group consisting of the sequence, the fragment, the epitope, the pool, the complex, the recombinant factor VIII or the inhibitor and/or the anti-inhibitor according to any one of the preceding paragraphs.
- a diagnostic and/or purification device which comprises at least one element which is selected from the group consisting of the sequence, the fragment, the epitope, the pool, the complex, the inhibitor and/or the anti-inhibitor according to any one of the preceding paragraphs.
- a method for a therapeutic treatment and/or prevention of an immune disorder in mammal wherein the pharmaceutical composition according to paragraph 22 is administered to the mammal patient presently or potentially having said immune disorder, in an amount effective to treat and/or prevent said immune disorder.
- a method for a therapeutic treatment and/or prevention of an immune disorder in a mammal patient wherein a physiological fluid such as serum obtained from said mammal patient is put into the chromatography column of paragraph 25 in order to allow a binding with the inhibitors of factor VIII present in said serum with the sequence, the fragment, the epitope, the pool and/or the complex according to any of the preceding paragraphs 1 to 15, wherein the physiological liquid is eluted from said chromatography column and the physiological liquid from which the inhibitors of factor VIII have been removed is reinjected to the patient.
- a physiological fluid such as serum obtained from said mammal patient is put into the chromatography column of paragraph 25 in order to allow a binding with the inhibitors of factor VIII present in said serum with the sequence, the fragment, the epitope, the pool and/or the complex according to any of the preceding paragraphs 1 to 15, wherein the physiological liquid is eluted from said chromatography column and the physiological liquid from which the inhibitors of factor VIII have been removed is reinjected to the patient.
- FIG. 1 This figure depicts the hydrophilicity, flexibility and accessibility graph of the A3 sequence of Factor VIII, renumbered 1 to 371 amino acids (surface value for each amino acid).
- FIG. 2 a represents the elution profile related to the purification of human anti-SEQ. ID NO: 32 antibodies by affinity chromatography in peptide-Sepharose column.
- Cohn fraction II+III solution 50 mL was loaded onto the column (1 mL gel) at a flow rate of 1 mL/min. The separation of specific antibodies was performed as described in the Examples.
- the arrow indicates the position of specific human anti-SEQ. ID NO: 32.
- the clotting activity of FVIII ( 2 b ) was measured as described in Examples in the presence of increasing amount of anti-SEQ. ID NO: 32.
- the of FVIII activity (FVIII activity in the presence of antibody/FVIII activity in the absence of antibody) * 100.
- FIG. 3 This figure represents the human anti-peptide antibody immunoreactions with FVIII polypeptides after Western Blotting (panel A from left to right: human antibodies HAP1 through HAP4, specific for different FVIII epitope sequences found in the FVIII HC—see also Table 2 and panel B: human antibodies specific for the P5 peptide and the FVIII LC sequences, P7, P8, and P9—see also Table 2).
- the RAP9 lane shows the reactivity of FVIII polypeptieds towards purified rabbit antibodies specific for the peptide sequence Arg 1797 -Tyr 1815 (see also Table 2).
- FIG. 4 This figure represents the ELISA reactivity of the four inhibitor plasmas with different peptide sequences. Inhibitors present in four patient plasmas were analyzed by ELISA test using as coated antigens the different selected FVIII epitopes synthetic peptides, as indicated by the ordinate.
- the present invention relates to the antigenic polypeptide sequences of factor VIII and/or fragments of these sequences, as described by Verhar et al. (1984), the disclosure of which is incorporated herein by reference in its entirety.
- polypeptide sequence of factor VIII is understood to be the natural human or animal sequence, which may be glycosylated and which has been obtained by purification from pools of plasma, in particular cryoprecipitate, by synthesis and/or by genetic manipulation (sequence from which portions which are not involved in the mechanism of blood coagulation may have been deleted) of factor VIII.
- the present invention relates, in particular, to the antigenic polypeptide sequences of factor VIII which lacks the fragments comprised between alanine 322-serine 750, leucine 1655-arginine 1689 and lysine 1694-proline 1782, and possibly also the fragments comprised between aspartic acid 2170 and tyrosine 2332.
- the present invention relates, in particular, to the antigenic polypeptide sequences A1, A2, A3 and C (C1 and C2) of factor VIII.
- a first embodiment of the invention relates to the antigenic polypeptide sequence A3 of factor VIII, and to fragments and/or epitopes of this sequence.
- the said sequence contains the fragments glutamic acid 1649 to histidine 2031 inclusive, arginine 1652 to arginine 1917 inclusive or arginine 1803 to arginine 1917 inclusive, of the polypeptide sequence of factor VIII as published by Verhar et al. (1984) and Toole et al. (1984).
- the fragments of the said sequence are arginine 1648 to arginine 1696 inclusive, threonine 1739 to aspartic acid 1831 inclusive or glutamic acid 1885 to arginine 1917 inclusive.
- the fragments, epitopes and major parts thereof are preferably polypeptidic sequences made of at least 7 amino acids of the FVIII polypeptidic sequence.
- the invention also relates to the sequence epitopes of these fragments, in particular:
- epitope arginine 1648 to tyrosine 1664 inclusive defined by the following sequence:
- epitope aspartic acid 1681 to arginine 1696 (P8) inclusive defined by the following sequence:
- epitope threonine 1739 to tyrosine 1748 inclusive defined by the following sequence:
- epitope asparagine 1777 to phenylalanine 1785 inclusive defined by the following sequence:
- epitope glutamic acid 1794 to tyrosine 1815 inclusive defined by the following sequence:
- epitope methionine 1823 to aspartic acid 1831 inclusive defined by the following sequence:
- epitope glutamic acid 1885 to alanine 1901 inclusive defined by the following sequence:
- epitope aspartic acid 1909 to arginine 1917 inclusive defined by the following sequence:
- epitope comprised between serine 2018 and histidine 2031 inclusive, defined by the following sequence:
- the said sequences, specific fragments and epitopes exhibit an antigenic characteristic which is illustrated by Table 1.
- Another preferred embodiment of the invention relates to antigenic polypeptide sequence Al of factor VIII, fragments and/or epitopes of this sequence.
- the fragments of the said sequence are alanine 108 to methionine 355 inclusive, preferably alanine 108 to alanine 227 inclusive.
- the invention also relates to the sequence epitopes of these fragments, in particular:
- epitope alanine 108 to valine 128 inclusive defined by the following sequence:
- epitope glutamic acid 181 to leucine 192 inclusive defined by the following sequence:
- epitope aspartic acid 203 to alanine 227 inclusive defined by the following sequence:
- epitope aspartic acid 327 to methionine 355 inclusive defined by the following sequence:
- Another preferred embodiment of the invention relates to the antigenic polypeptide sequence A2 of factor VIII, fragments and/or epitopes of this sequence.
- the fragments of the said sequence are aspartic acid 403 to serine 840 inclusive, preferably histidine 693 to aspartic acid 725 inclusive.
- the invention also relates to the sequence epitopes of these fragments, in particular:
- epitope aspartic acid 403 to lysine 425 inclusive defined by the following sequence:
- epitope histidine 693 to glycine 701 inclusive defined by the following sequence:
- epitope serine 710 to aspartic acid 725 inclusive defined by the following sequence (P4):
- a final preferred embodiment of the invention relates to the antigenic polypeptide sequence C of factor VIII, and fragments and/or epitopes of this sequence.
- the fragments of the said sequence are histidine 2082 to lysine 2251 inclusive or leucine 2273 to tyrosine 2332 inclusive, preferably lysine 2085 to glycine 2121 inclusive and serine 2181 to leucine 2251 inclusive.
- the invention also relates to the sequence epitopes of these fragments, in particular:
- epitope isoleucine 2081 to serine 2095 inclusive defined by the following sequence:
- epitope tyrosine 2105 to glycine 2121 inclusive defined by the following sequence:
- epitope asparagine 2128 to asparagine 2138 inclusive defined by the following sequence:
- epitope histidine 2152 to arginine 2163 inclusive defined by the following sequence:
- epitope serine 2181 to asparagine 2198 inclusive defined by the following sequence:
- epitope glycine 2242 to leucine 2251 inclusive defined by the following sequence:
- epitope isoleucine 2262 to glutamine 2270 inclusive defined by the following sequence:
- epitope proline 2292 to tyrosine 2305 inclusive defined by the following sequence (P15):
- the invention also relates to the major parts of the said epitopes or the said fragments.
- Said epitopes can be deleted from one or more terminal amino acids, preferably from one, two or three amino acids, or can be replaced by one or more amino acids that present the same characteristic of hydrophilicity, flexibility and accessibility.
- epitopes according to the invention are comprised in major determinants of human inhibitors epitopes or several factors binding sites or binding sites of known monoclonal antibodies, especially the portion C2 that is known to be the binding site of the monoclonal antibody Mas531P or the binding site ESH8 as well as phospholipids, Factor Xa or the von Willebrand factor binding site.
- the specific epitopes according to the invention or their major parts are preferred selected portions of said binding sites or may include a possible overlapping with said binding sites.
- the epitopes according to the invention are more specific portions of known epitopes. Therefore, an artificial epitope could be easily obtained by synthesis and the specific above-described fragments can be deleted from non-epitopic portions such as the fragment described in a C2 fragment (amino acids phenylalanine 2196 to tryptophan 2203 inclusive and amino acids valine 2222 to phenylalanine 2234 inclusive, or the sequence leucine 2252 to threonine 2272 inclusive or the amino acids phenylalanine 2290 to threonine 2291 inclusive as well as the amino acids leucine 2306 to methionine 2321 inclusive).
- Another aspect of the present invention is related to a modified (recombinant or transgenic) FVIII, possibly obtained by genetic engineering, and deleted from one or more of the above-identified fragments, epitopes or major parts of said epitopes and/or said fragments.
- said FVIII still allows the binding of coagulation factor(s), but will be less immunogenic and will not induce or induce less the formation of inhibitors directed against said modified FVIII or natural FVIII.
- said polypeptide sequences, fragments or epitopes are also independently immunogenic (that is to say they are immunogenic even without being complexed with a protein of large size such as BSA, KLH haemocyanin, etc.), and preferably exhibit an immunoaffinity within inhibitors of factor VIII, such as anti-factor VIII antibodies, and/or exhibit an immunoaffinity for the receptors of the T lymphocytes and possibly B lymphocytes.
- Said sequences are unexpectedly characterized by substantial immunogenicity towards monoclonal and polyclonal antibodies, but are sufficiently short to be readily and advantageously obtained by synthesis.
- the present invention also relates to the conformational epitopes which comprise at least two different fragments of said sequence, at least two sequence epitopes and/or at least two major parts of said epitopes or said different fragments according to the invention and above identified.
- the conformational epitopes are made up of two or more different portions of a polypeptide sequence, which portions are located in proximity to each other when the protein is folded in its tertiary or quaternary structure.
- These epitopes are capable of being “recognized” (that is to say of exhibiting an immunoaffinity), preferably simultaneously, with inhibitors of factor VIII, in particular B and T lymphocytes (by way of the major histocompatibility locus (MHC I and/or II)) and/or anti-factor VIII antibodies (Scandella et al. (2000); Reding et al. (2000)).
- the said sequence, said fragments, said epitopes and/or the major parts of said epitopes or said fragments are complexed with a carrier protein or a carrier peptide, such as BSA, or KLH haemocyanin, as to form a complex exhibiting a more powerful immunogenicity.
- a carrier protein or a carrier peptide such as BSA, or KLH haemocyanin
- the present invention is also related to a pool of more than three of said fragments, epitopes or major parts of said epitopes having advantageously important antigenic and/or immunogenic properties and which may be used advantageously in a diagnostic or therapeutic method or device such as a kit or a dialysis column, etc. allowing an efficient, preferably complete, screening and characterization of the major (if not all) known inhibitors directed against factor VIII by (human) patients.
- Another aspect of the present invention relates to an inhibitor of factor VIII which exhibits an immunoaffinity with antigenic polypeptide sequences according to the present invention, with fragments and epitopes of said sequences, with the major parts of said epitopes or said fragments and/or with the complex according to the invention.
- An inhibitor is understood to mean any biological molecule or cell (such as a T-lymphocyte) binding to said FVIII and capable of giving rise to immune disorders (characterized by humoral immune response and/or cellular immune response against said FVIII).
- such an inhibitor can be an anti-factor VIII monoclonal or polyclonal antibody or antibody fragment (such as the hypervariable Fab portion of the said antibody) which inactivates the said factor VIII and/or which inhibits the binding of factor VIII to the von Willebrand factor and/or to membrane phospholipids.
- an anti-factor VIII monoclonal or polyclonal antibody or antibody fragment such as the hypervariable Fab portion of the said antibody
- the said inhibitors are synthesized by a “chimeric” animal which comprises a human immune system, such as an hu-SCID mouse or transgenic mouse producing human antibodies or other antibodies production technologies as phage display technology or immortalized B-cells, by EPV in particular.
- a human immune system such as an hu-SCID mouse or transgenic mouse producing human antibodies or other antibodies production technologies as phage display technology or immortalized B-cells, by EPV in particular.
- Another aspect of the invention relates to an anti-inhibitor which is directed against the said previously described factor VIII inhibitor.
- An anti-inhibitor which is directed against the factor VIII inhibitor is understood to mean any chemical or biological molecule, a cell and/or a cell fragment (receptor) which is capable of interfering with the said inhibitor in such a way as to ensure its inactivation or avoid or reduce its binding to the factor VIII.
- such an anti-inhibitor is an anti-anti-factor VIII idiotype (monoclonal or polyclonal) antibody or antibody fragment, natural or obtained by genetic engineering.
- Another aspect of the invention relates to a pharmaceutical composition which comprises an adequate pharmaceutical carrier or a diluent and an element selected from the group consisting of said antigenic polypeptide sequence of factor VIII, fragments and epitopes of this sequence or a pool thereof, an inhibitor of factor VIII which is directed against them, an anti-inhibitor which is directed against the said inhibitor, and/or a mixture of these.
- the type and amount of adequate pharmaceutical carrier or diluent (and possibly adjuvant or excipient) present in said pharmaceutical composition may vary according to the method of administration and is possibly combined an adjuvant in order to improve therapeutical properties of the pharmaceutical composition according to the invention or to reduce its possible side effects.
- Suitable pharmaceutical acceptable carriers used in the pharmaceutical composition according to the invention are well known by the person skilled in the art and are selected according to the methods generally applied by pharmacists and may include solid, liquid or gaseous non-toxic pharmaceutically acceptable carriers.
- the percentage of active product/pharmaceutical acceptable carrier may vary within very large ranges only limited by the tolerance and the possible side effects on patients (including humans), and by frequency and/or mode of administration.
- a diagnostic and/or purification device such as a diagnostic kit, an affinity filter, or a chromatography column which comprises an element which is selected from the group consisting of these antigenic polypeptide sequences, fragments and epitopes and/or major parts of said epitopes or said fragments, the complex according to the invention or a pool thereof, an inhibitor which is directed against them, an anti-inhibitor which is directed against said inhibitor, and/or a mixture of these.
- said device comprises a pool of said epitopes which allow a screening of patients and may detect the most important inhibitors present in said patients and which allow a positive test with enough specificity and sensibility.
- the purification device can therefore consist of a chromatography column which comprises these sequences of factor VIII, fragments and epitopes and/or major parts of said fragments or epitopes, attached to the solid phase of the chromatography column.
- a physiological liquid (such as serum), which is derived from a patient and which comprises inhibitors of factor VIII pass through this chromatography column, with said inhibitors (for example antibodies) becoming attached specifically to said factor VIII sequences, fragments, epitopes or said major parts or a pool thereof. Following elution, it is possible to collect said inhibitors by causing them to react with anti-inhibitors (anti-anti-factor VIII idiotype antibodies).
- the present invention is also related to a method of treatment (ex vivo treatment) of a patient suffering from a pathology induced by inhibitors to the factor VIII which comprises the steps of extracting said physiological liquid (blood or serum) from the patient, obtaining its reaction upon a solid support binding the factor VIII fragments, epitopes or a pool thereof according to the invention and reinjecting said physiological liquid to the patient after the removing of the inhibitors having fixed said factor VIII fragments, epitopes, majors parts or a pool thereof.
- a method of treatment ex vivo treatment of a patient suffering from a pathology induced by inhibitors to the factor VIII which comprises the steps of extracting said physiological liquid (blood or serum) from the patient, obtaining its reaction upon a solid support binding the factor VIII fragments, epitopes or a pool thereof according to the invention and reinjecting said physiological liquid to the patient after the removing of the inhibitors having fixed said factor VIII fragments, epitopes, majors parts or a pool thereof.
- a final aspect of the invention relates to the use of the pharmaceutical composition according to the invention for preparing a medicament used for preventing and/or treating immune disorders, in particular those induced by inhibitors of factor VIII, inhibitors of the binding of factor VIII to the factor IX and/or the factor X and/or the von Willebrand factor (vWF) and/or inhibitors of the binding of factor VIII to membrane phospholipids.
- immune disorders in particular those induced by inhibitors of factor VIII, inhibitors of the binding of factor VIII to the factor IX and/or the factor X and/or the von Willebrand factor (vWF) and/or inhibitors of the binding of factor VIII to membrane phospholipids.
- vWF von Willebrand factor
- Reagents MAS530p (Harlan-Seralab, Indianapolis, Ind.) is a mouse monoclonal antibody specific for the 44-kDa A2 domain of the factor VIII heavy chain. Biotin-labeled rabbit IgG anti-mouse IgG was purchased from Dakopatts (Copenhagen, Denmark).
- Biotin-labeled goat IgG anti-human IgG and biotin-labeled mouse IgG anti-rabbit IgG were obtained from Sigma Chemicals (St Louis, Mich.), purified a-thrombin (3000 IU/mg), streptavidin-peroxidase conjugate, ovalbumin (OVA), bovine serum albumin (BSA), keyhole limpet haemocyanin (KLH), and o-phenylenediamine (OPD) were purchased from Sigma Chemicals (St. Louis, Mich.). Casein was obtained from Merck (Darmstadt, Germany). 4-chloro-1-naphtol and biotinylated molecular weight markers were obtained from Bio-Rad Laboratories (Hercules, Calif.). Freund's adjuvant was from Difco (Detroit, Mich.).
- Plasma FVIII was a solvent/detergent-treated FVIII concentrate (100 IU/mg protein) purified by ion exchange chromatography (FVIII Conc. SD, CAF-DCF-Red Cross, Brussels, Belgium). Albumin-free recombinant FVIII (rFVIII) was obtained from Hyland (Glendale, Calif.).
- Plasma fraction immunoglobulins Cohn Fraction II+III was obtained from large plasma pool from 4,800 unpaid donors, after precipitation in the presence of increasing ethanol concentration. This fraction contains all Ig classes and subclasses. IgG composition was determined by nephelometry. The relative percentage of each subclass was 63,7; 30,1; 3,4 and 2,8 for IgG1, IgG2, IgG3 and IgG4 respectively (average values for 3 different batches of FII+III).
- Factor VIII activity was determined in a one-stage clotting assay adapted for use on the Coagulometer KC4A (Sigma Diagnostics). The assay uses severe hemophilia A plasma (Organon Teknika, Cambridge, UK) and APPT reagent from Instrumentation Laboratory (Warrington, UK). Potencies were calculated relative to the 5 th International Standard FVIII concentrate 88/640 (5.4 IU/ml) (NIBSC, Potters Bar, UK). FVIII-inhibitory activity was measured in purified rabbit and human IgG preparations according to the modified Bethesda assay. Briefly, affinity-purified IgGs were serially diluted and incubated for 1 h in the presence of FVIII concentrate 88/640 (1 IU/ml) at 37° C. The residual FVIII activity was measured as described above.
- samples were dialyzed 3 times against 5 volumes of TE buffer (20 mM Tris-HCl pH 7.2, 150 mM NaCl and 0.02% NaN 3 ) and loaded onto the column at a flow rate of 1 ml/min.
- the column was sequentially washed at 2 ml/min with 50 ml TE buffer and 30 ml TE containing 1 M NaCl.
- the material was eluted (1 ml/min) with 5 ml of 0.1 M citric acid pH 2.5 and directly recovered in 5 ml of IM Tris-HCl, pH 9.0.
- Samples were finally dialyzed versus 10 volumes of equilibration buffer and concentrated on Centriprep-30 (Amicon, Beverly, Mass.). Ig recovery was determined by the Bio-Rad protein assay.
- Results are summarized in Table 1 which concerns the characterization of rabbit anti-FVIII-peptide antisera and recovered affinity-purified of immunoglobulins.
- Sixteen synthetic peptides (from 10 to 20 amino acids) were selected in the A, B, Cl and C2 domains. After conjugation with ovalbumin, the OVA-peptide conjugates were injected into rabbits and FVIII anti-peptide antisera RAP1 to RAP16 were studied. More precisely, two rabbits were immunized with each FVIII-peptide-ovalbumin preparation.
- RAP1 to RAP16 were prepared and assayed in an ELISA (column c, Table 1) using rFVIII or FVIII-peptide-KLH as the antigen.
- ELISA titer is expressed as the negative log of the reciprocal of the serum dilution giving 50% binding.
- the immunoglobulins were then purified by chromatography on peptide-bound Sepharose.
- the FVIII domain recognized by the anti-FVIII peptide Ig after immunoblotting is shown in (column d, Table 1) and Ig protein recoveries (column e, Table 1) were measured using immunoglobulins as the standard.
- the inhibitory activity, expressed in BU/mg protein, was determined in a FVIII neutralizing activity assay (column f, Table 1).
- FVIII anti-peptide antisera The reactivity of FVIII anti-peptide antisera was measured by an ELISA using, as antigen, either the different corresponding FVIII-peptide coupled to KLH protein or purified rFVIII.
- the binding reaction of each anti-FVIII-peptide antiserum was specific both for the FVIII peptide used to elicit the immune response in rabbit and for rFVIII (see Table 1).
- the specific rabbit IgG were purified by affinity chromatography on peptide-Sepharose as described under Methods. When FVIII-neutralizing activity was measured in a one-stage clotting assay, significant inhibition was found with two rabbit IgG purified preparations: RAP2, corresponding to IgG specific for SEQ ID No. 14 and RAP7 specific for SEQ ID NO.:1.
- rFVIII and the rFVIII fragments obtained after treatment with thrombin were resolved by SDS-PAGE and analyzed by western blotting with different preparations of rabbit IgGs (RAP1 to RAP17 Igs).
- Antisera RAP1 and RAP2 reacted with the 50-kDa A1-domain fragment; RAP3 and RAP4 bound to the 44-kDa fragment (domain A2); RAP5 (specific for the B domain) bound to the high-molecular-weight FVIII heavy chain (about 200-kDa).
- RAP7, RAP8, and RAP9 reacted with the 80-kDa light-chain doublet.
- RAP9 and RAP12 to RAP17 antibodies also detected the 72-kDa FVIII light-chain fragment.
- each reactive antiserum showed a strong reaction with the corresponding FVIII fragment containing the selected linear epitope. No reaction was detectable in the gels between RAP6 or RAP10 and the HC or LC FVIII fragments.
- Table 2 concerns the characterization of human anti-FVIII antibodies from Cohn fraction II+III of healthy individuals.
- FIG. 2 shows the chromatographic profile obtained with SEQ ID 32, a sequence found in C2 domain. Table 2 summarizes the results obtained with 17 epitopic sequences selected in each FVIII domain (Al, A2, A3, B, Cl and C2).
- IgG isotype distribution in the human purified antibody preparations was found to be quite heterogeneous. Interestingly, 40 to 79% of the recovered IgGs belonged to the IgG2 subclass. In most preparations, IgG4 appeared to be over-represented (up to 25%).
- FIG. 3 shows the immunoreaction of high-molecular-weight FVIII ( ⁇ 92-kDa) with four human antibody preparations, purified on Sepharose coupled to FVIII peptide SEQ ID NO.: 11 (Ser 109 -Lys 127 ), SEQ ID NO.: 14 (Cys 329 -Asp 348 ), SEQ ID NO.: 15 (Tyr 407 -Lys 425 ) or SEQ ID NO.: 19 (Cys 711 -Asp 725 ).
- the 50-kDa FVIII fragment (domain A1) was recognized by human antibodies purified on Ser 109 -Lys 127 or Cys 329 -Asp 348 -Sepharose and the 44-kDa FVIII fragment (A2) by immunoglobulins purified on Tyr 407 -Lys 425 and Cys 711 -Asp 725 -Sepharose.
- the lack of reactivity of the anti-(Ser 817 -Ser 830 ) immunoglobulin preparation (HAP5) with the FVIII fragments confirms that this epitope is located in the amino-terminal end of domain B (FIG. 3).
- the selected peptides were used in ELISA experiment to determine the anti-FVIII antibody specificity's present in hemophilia A plasmas.
- the peptides were coated on microplate (25 ⁇ g/ml in PBS buffer during 16 h at 4° C.).
- a ⁇ fraction (1/10) ⁇ to ⁇ fraction (1/1000) ⁇ dilution of plasma patient in Tris-casein buffer was reacted with the coated peptide for 2 h at 37° C.
- the bound human IgG was measured as described in Methods.
- Control samples were plasma pools of healthy donors.
- FIG. 4 shows the results obtained with the plasma of 4 hemophilia A patients.
- the optical densities are corrected average values (OD patient-OD normal plasma pool) of two independent experiments.
- Epitope Arg 1652 -Tyr 1664 and the adjacent major binding site vWF are located in the acidic light-chain peptide a3. As shown by western blotting, a3 is released from the A3 domain after thrombin treatment, preventing further binding of anti-(Arg 1652 -Tyr 1664 ) Ig to activated FVIII. Similar results have been reported by Shima et al (1991), who described the FVIII sequence Asp 1663 -Ser 1669 as a binding site of rabbit polyclonal antibodies neutralizing FVIII activity.
- Epitope Cys 329 -Asp 348 overlapped the acidic Asp 348 -Lys 362 sequence (in al) described as adjacent to the activated protein C (Arg 336 ) and thrombin (Arg 372 ) cleavage sites. It is the target of human hemophilic inhibitors.
- Anti-(Asp 348 -Lys 362 ) antibodies may interfere with proteolysis or with the FX interaction site (Met 337 -Arg 372 ) (Saenko et al., 1999 and Scandella et al., 2000).
- FVIII-neutralizing activity was measured in all 13 Ig preparations. Seven human Ig preparations displayed inhibition of procoagulant activity, these being specific for amino-acid residues Cys 711 -Asp 725 , Tyr 1681 -Arg 1696 and Arg 1797 -Tyr 1815 respectively.
- the Cys 711 -Asp 725 sequence contains sulfated tyrosines at Tyr 718 , Tyr 719 , and Tyr 723 , and overlaps with the FVIII HC region Lys 713 -Arg 740 described as promoting both activation and HC proteolysis.
- the additional sulfated groups may be required for proper interaction with thrombin or another component as in the FX-activating complex.
- Peptide P8 (Tyr 1681 -Arg 1696 ) (FVIII LC) includes the sequence Glu 1684 -Arg 1689 already described by Shima et al, 1991. It contains the thrombin activation site Arg 1689 -Ser 1690 .
- P4 (Cys 711 -Asp 725 ) is also included in the Asp 712 -Ala 736 sequence detected by analysis of the patient antibody repertoire by gene phage display technology. It is proposed as a possible additional inhibitor in patients (van den Brink et al, 2000).
- Peptide P9 (Arg 1797 -Tyr 1815 ) contains the FXa binding site (see below).
- FVIII epitope sequences help to determine the contribution of patient polyclonal anti-FVIII Igs to overall inhibitory and regulatory activity. They could also be used to monitor the usual switch in anti-FVIII specificity in a patient during treatment. Said characterization of FVIII epitopes and a model of their locations on the folded molecule improves the treatment of inhibitors in both hemophilic and non-hemophilic patients (detection, follow-up, therapeutic use of FVIII epitope peptides . . . ).
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Hematology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Urology & Nephrology (AREA)
- Microbiology (AREA)
- Genetics & Genomics (AREA)
- Animal Behavior & Ethology (AREA)
- Biophysics (AREA)
- Food Science & Technology (AREA)
- Cell Biology (AREA)
- Pathology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Mycology (AREA)
- General Chemical & Material Sciences (AREA)
- Toxicology (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Rheumatology (AREA)
- Biotechnology (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Diabetes (AREA)
- Peptides Or Proteins (AREA)
Priority Applications (8)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US09/853,080 US20020068303A1 (en) | 1994-07-14 | 2001-05-09 | Antigenic polypeptide sequences of factor VIII, and fragments and/or epitopes of these sequences |
AT02732243T ATE525470T1 (de) | 2001-05-09 | 2002-05-06 | Antigene epitope des faktors viii, inhibitoren dagegen, und deren verwendungen |
CA2446390A CA2446390C (en) | 2001-05-09 | 2002-05-06 | Antigenic epitopes of factor viii, inhibitors directed against said epitopes and use thereof |
PCT/BE2002/000070 WO2002090542A2 (en) | 2001-05-09 | 2002-05-06 | Antigenic epitopes of factor viii, inhibitors directed against said epitopes and use thereof |
ES02732243T ES2372811T3 (es) | 2001-05-09 | 2002-05-06 | Epítopos antigénicos del factor viii, inhibidores dirigidos contra dichos epítopos y uso de los mismos. |
EP02732243A EP1399556B1 (en) | 2001-05-09 | 2002-05-06 | Antigenic epitopes of factor viii, inhibitors directed against said epitopes and use thereof |
JP2002587603A JP4500494B2 (ja) | 2001-05-09 | 2002-05-06 | 第viii因子の抗原エピトープ、当該エピトープに対して向けられたインヒビターおよびそれらの使用 |
US11/267,631 US7981865B2 (en) | 1994-07-14 | 2005-11-03 | Antigenic fragments of human factor VIII polypeptides |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
BE09400666 | 1994-07-14 | ||
BE9400666A BE1008491A3 (fr) | 1994-07-14 | 1994-07-14 | Sequence polypeptidique antigenique du facteur viii, fragments et/ou epitopes de celle-ci. |
US08/765,837 US6866848B2 (en) | 1994-07-14 | 1995-07-14 | Antigenic polypetide sequence of factor VIII, fragments and/or epitopes there of |
US09/853,080 US20020068303A1 (en) | 1994-07-14 | 2001-05-09 | Antigenic polypeptide sequences of factor VIII, and fragments and/or epitopes of these sequences |
Related Parent Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/BE1995/000068 Continuation-In-Part WO1996002572A2 (fr) | 1994-07-14 | 1995-07-14 | Sequence polypeptidique antigenique du facteur viii, fragments et/ou epitopes de celle-ci |
US08/765,837 Continuation-In-Part US6866848B2 (en) | 1994-07-14 | 1995-07-14 | Antigenic polypetide sequence of factor VIII, fragments and/or epitopes there of |
US08765837 Continuation-In-Part | 1995-07-14 |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/267,631 Continuation US7981865B2 (en) | 1994-07-14 | 2005-11-03 | Antigenic fragments of human factor VIII polypeptides |
Publications (1)
Publication Number | Publication Date |
---|---|
US20020068303A1 true US20020068303A1 (en) | 2002-06-06 |
Family
ID=25314986
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/853,080 Abandoned US20020068303A1 (en) | 1994-07-14 | 2001-05-09 | Antigenic polypeptide sequences of factor VIII, and fragments and/or epitopes of these sequences |
US11/267,631 Expired - Fee Related US7981865B2 (en) | 1994-07-14 | 2005-11-03 | Antigenic fragments of human factor VIII polypeptides |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/267,631 Expired - Fee Related US7981865B2 (en) | 1994-07-14 | 2005-11-03 | Antigenic fragments of human factor VIII polypeptides |
Country Status (7)
Country | Link |
---|---|
US (2) | US20020068303A1 (es) |
EP (1) | EP1399556B1 (es) |
JP (1) | JP4500494B2 (es) |
AT (1) | ATE525470T1 (es) |
CA (1) | CA2446390C (es) |
ES (1) | ES2372811T3 (es) |
WO (1) | WO2002090542A2 (es) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
BR0308860A (pt) * | 2002-04-18 | 2005-01-04 | Merck Patent Gmbh | Fator viii modificado |
GB0713187D0 (en) * | 2007-07-06 | 2007-08-15 | Cambridge Entpr | biomolecule binding ligands |
GB0801513D0 (en) * | 2008-01-28 | 2008-03-05 | Circassia Ltd | Peptides from factor VIII |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4965199A (en) * | 1984-04-20 | 1990-10-23 | Genentech, Inc. | Preparation of functional human factor VIII in mammalian cells using methotrexate based selection |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4649132A (en) | 1983-03-31 | 1987-03-10 | Scripps Clinic And Research Foundation | Treatment of Factor VIII inhibitors |
KR910006424B1 (ko) | 1985-08-21 | 1991-08-24 | 인코텍스 비.브이 | 편성브리프(brief) 제조방법 |
BE1008491A3 (fr) * | 1994-07-14 | 1996-05-07 | Croix Rouge De Belgique Depart | Sequence polypeptidique antigenique du facteur viii, fragments et/ou epitopes de celle-ci. |
EP0965597A4 (en) * | 1996-12-27 | 2003-01-08 | Mochida Pharm Co Ltd | CELL MEMBRANE EFFECTIVE DRUGS |
EP1051432B1 (en) * | 1998-12-08 | 2007-01-24 | Biovation Limited | Method for reducing immunogenicity of proteins |
AU2002249779A1 (en) * | 2000-12-01 | 2002-08-12 | Regents Of The University Of Minnesota | Method to treat hemophilia |
-
2001
- 2001-05-09 US US09/853,080 patent/US20020068303A1/en not_active Abandoned
-
2002
- 2002-05-06 EP EP02732243A patent/EP1399556B1/en not_active Expired - Lifetime
- 2002-05-06 JP JP2002587603A patent/JP4500494B2/ja not_active Expired - Fee Related
- 2002-05-06 CA CA2446390A patent/CA2446390C/en not_active Expired - Fee Related
- 2002-05-06 WO PCT/BE2002/000070 patent/WO2002090542A2/en active Application Filing
- 2002-05-06 ES ES02732243T patent/ES2372811T3/es not_active Expired - Lifetime
- 2002-05-06 AT AT02732243T patent/ATE525470T1/de active
-
2005
- 2005-11-03 US US11/267,631 patent/US7981865B2/en not_active Expired - Fee Related
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4965199A (en) * | 1984-04-20 | 1990-10-23 | Genentech, Inc. | Preparation of functional human factor VIII in mammalian cells using methotrexate based selection |
Also Published As
Publication number | Publication date |
---|---|
ES2372811T3 (es) | 2012-01-26 |
US20060051367A1 (en) | 2006-03-09 |
WO2002090542A8 (en) | 2004-02-12 |
EP1399556B1 (en) | 2011-09-21 |
JP4500494B2 (ja) | 2010-07-14 |
EP1399556A2 (en) | 2004-03-24 |
WO2002090542A2 (en) | 2002-11-14 |
CA2446390C (en) | 2012-01-17 |
US7981865B2 (en) | 2011-07-19 |
WO2002090542A3 (en) | 2003-12-24 |
JP2004535393A (ja) | 2004-11-25 |
ATE525470T1 (de) | 2011-10-15 |
CA2446390A1 (en) | 2002-11-14 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Gilles et al. | Anti-factor VIII antibodies of hemophiliac patients are frequently directed towards nonfunctional determinants and do not exhibit isotypic restriction | |
Gilles et al. | Healthy subjects produce both anti-factor VIII and specific anti-idiotypic antibodies. | |
Gilles et al. | Neutralizing antiidiotypic antibodies to factor VIII inhibitors after desensitization in patients with hemophilia A. | |
Nogami et al. | A novel mechanism of factor VIII protection by von Willebrand factor from activated protein C–catalyzed inactivation | |
AU626255B2 (en) | Factor viii binding domain of von willebrand factor | |
JP2009528285A (ja) | ファクターviiiのc1ドメインに対する阻害抗体の阻害活性を中性化する抗イデオタイプ抗体 | |
JPH0638790A (ja) | ファクターviii:c凝固因子ポリペプチド類に特異的なモノクローナル抗体 | |
JP2006316062A (ja) | 因子viiiの抗原ポリペプチド配列、その断片および/またはエピトープ | |
ES2377128T3 (es) | Anticuerpo antiidiotípico que neutraliza la actividad inhibidora de un anticuerpo inhibidor del factor VIII | |
Gilles et al. | Some factor VIII (FVIII) inhibitors recognise a FVIII epitope (s) that is present only on FVIII-vWF complexes | |
US7981865B2 (en) | Antigenic fragments of human factor VIII polypeptides | |
EP0319315A2 (en) | The von Willebrand factor binding domain of factor VIII | |
Mühle et al. | Epitope mapping of polyclonal clotting factor VIII-inhibitory antibodies using phage display | |
Shima et al. | Localization of the binding site for a factor VIII activity neutralizing antibody to amino acid residues Asp1663-Ser1669. | |
US7582296B2 (en) | Anti-idiotypic antibodies against factor VIII inhibitor and uses thereof | |
Shima | Characterization of factor VIII inhibitors | |
Gharagozlou et al. | Epitope specificity of anti-factor VIII antibodies from inhibitor positive acquired and congenital haemophilia A patients using synthetic peptides spanning A and C domains | |
Di Giambattista et al. | Mapping of natural anti-factor VIII antibodies in plasma pools from healthy donors: use of rationally designed synthetic peptides | |
Di Giambattista et al. | In silico prediction of FVIII epitopes recognised by natural autoantibodies in polyvalent immunoglobulin concentrates | |
Saenko et al. | Molecular modifications in factor VIII concentrates produced from different plasma pools | |
Furlong et al. | Monoclonal antibodies to factor VIII: their application in immunoblotting for the visualization of factor VIII in therapeutic concentrates and plasma | |
Gilles et al. | Restricted epitope specificity of anti-FVIII antibodies that appeared during a recent outbreak of inhibitors | |
MÜHLE et al. | and H. SCHNEIDER | |
Mühle et al. | Characterization of Polyclonal Factor VIII-Inhibitory Antibodies |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: DEPARTEMENT CENTRAL DE FRACTIONNEMENT DE LA CROIX- Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:LAUB, RUTH;DI GIAMBATTISTA, MARIO;REEL/FRAME:012200/0049 Effective date: 20010615 |
|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |