Method for solid phase peptide synthesis
AssigneePolypeptide Laboratories Holding (PPL) AB
InventorsAnne-Sophie Droz, Jasmine Schnidrig, Nicole Studer, Stéphane Varray, Corinne Wenger, Oleg Werbitzky
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Abstract
A novel method for synthesizing a Hirulog peptide is devised.
Claims42
A peptide-resin conjugate A-W, wherein A=P-X1-Asp(R4)-Phe-Glu(R5)-Glu(R6)-Ile-Pro-Glu(R7)-Glu(R8)-Tyr(R9)-X2 (SEQ ID NO:3), wherein X1 is a peptidyl moiety of 0 to 200 amino acids, X1 optionally comprising protection groups on individual amino acid side chains, wherein R9 is an amino side chain protection group and wherein X2 is a single amino acid residue linked to the solid phase via —O— and optionally being side chain or C-terminally protected, and wherein P is H or is a protection group selected from the group consisting of Boc, Fmoc, Dde, Nps, Alloc, Z, and R4, R5, R6, R7 and R8 are amino acid side chain protection groups, and
- wherein W is a solid phase composite comprising a resin handle or linker
- a) of the formula II
- with the proviso that then when A, where including includes a residue X2, A is always linked via —O— to said handle or linker,
- and wherein R′″ is a solid phase, and wherein R″1, and R″2, R″3 are, independently, H, 4-(C1-C4 alkyl) or 4′-(C1-C4 alkyl) or 4-(C1-C4 alkoxy) or 4′-(C1-C4 alkoxy), and may be are the same or different with the proviso that only one of R″1, R″2 may can be H, and wherein R″2 may optionally be 2-Cl with the proviso that then when R″1 is H,
- b) or of the formula III
- with the proviso that then when A, where including includes a residue X2, A is linked via —O— to said handle or linker, and R′″ being defined as above is a solid phase,
- c) or of the formula IV
- wherein R′″ is defined as above and R″1, R″2, R″3 are, independently, H, C1-C4 alkyl or C1-C4 alkoxy, and may be are the same or different with the provisio proviso that only one of R″1, R″2 may can be H, and
- wherein L is A(L=A) or wherein L is of formula V
- and wherein W allows of cleaving the peptide moiety under weakly acidic conditions of 0.1% to 30% trifluoroacetic acid.
A process of using a peptide resin conjugate A-W for the synthesis of Bivalirudin, the process comprising
- cleaving a protected peptide from the peptide resin conjugate A-W,
- wherein
- A=Boc-D-Phe-Pro-Arg(R2)-Pro-Gly-Gly-Gly-Gly-Asn(R3)-Gly-Asp(R4)-Phe-Glu(R5)-Glu(R6)-Ile-Pro-Glu(R7)-Glu(R8)-Tyr(R9)-Leu-O— or
- A=Fmoc-D-Phe-Pro-Arg(R2)-Pro-Gly-Gly-Gly-Gly-Asn(R3)-Gly-Asp(R4)-Phe-Glu(R5)-Glu(R6)-Ile-Pro-Glu(R7)-Glu(R8)-Tyr(R9)-Leu-O— or
- A=NH2-D-Phe-Pro-Arg(R2)-Pro-Gly-Gly-Gly-Gly-Asn(R3)-Gly-Asp(R4)-Phe-Glu(R5)-Glu(R6)-Ile-Pro-Glu(R7)-Glu(R8)-Tyr(R9)-Leu-O—
- wherein R2, R3, R4, R5, R6, R7, R8, R9 are amino side chain protection groups, and
- wherein W is a solid phase composite comprising a resin handle or linker
- a) of the formula II
- with the proviso that when A includes a residue X2, A is linked via —O— to said handle or linker,
- wherein R′″ is a solid phase, and wherein R″1 and R″2 are independently, H, 4-(C1-C4 alkyl) or 4′-(C1-C4 alkyl) or 4-(C1-C4 alkoxy) or 4′-(C1-C4 alkoxy), and are the same or different with the proviso that only one of R″1, R″2 can be H, and wherein R″2 may optionally be 2-Cl when R″1 is H,
- b) or of the formula III
- with the proviso that when A includes a residue X2, A is linked via —O— to said handle or linker and R′″ is a solid phase,
- c) or of the formula IV
- wherein R′″ is defined as above and R″1, R″2, R″3 are, independently, H, C1-C4 alkyl or C1-C4 alkoxy, and are the same or different with the proviso that only one of R″1, R″2 can be H, and
- wherein L is A(L=A) or wherein L is of formula V
- and wherein W allows of cleaving the peptide moiety under weakly acidic conditions of 0.1% to 30% trifluoroacetic acid.
A process of using a peptide resin conjugate A-W for the synthesis of a Hirulog peptide, the process comprising
- cleaving a protected peptide from the peptide resin conjugate A-W, and
- deprotecting the protected peptide,
- wherein A=P-X1-Asp(R4)-Phe-Glu(R5)-Glu(R6)-Ile-Pro-Glu(R7)-Glu(R8)-Tyr(R9)-X2 (SEQ ID NO:3), wherein X1 is a peptidyl moiety of 0 to 200 amino acids, X1 optionally comprising protection groups on individual amino acid side chains, wherein R9 is an amino side chain protection group and wherein X2 is a single amino acid residue linked to the solid phase via —O— and optionally being side chain or C-terminally protected, and wherein P is H or is a protection group selected from the group consisting of Boc, Fmoc, Dde, Nps, Alloc, Z, and R4, R5, R6, R7 and R8 are amino acid side chain protection groups, and
- wherein W is a solid phase composite comprising a resin handle or linker
- a) of the formula II
- with the proviso that when A includes a residue X2, A is linked via —O— to said handle or linker,
- wherein R′″ is a solid phase, and wherein R″1 and R″2 are independently, H, 4-(C1-C4 alkyl) or 4′-(C1-C4 alkyl) or 4-(C1-C4 alkoxy) or 4′-(C1-C4 alkoxy), and are the same or different with the proviso that only one of R″1, R″2 can be H, and wherein R″2 may optionally be 2-Cl when R″1 is H,
- b) or of the formula III
- with the proviso that when A includes a residue X2, A is linked via —O— to said handle or linker and R′″ is a solid phase,
- c) or of the formula IV
- wherein R′″ is defined as above and R″1, R″2, R″3 are, independently, H, C1-C4 alkyl or C1-C4 alkoxy, and are the same or different with the proviso that only one of R″1, R″2 can be H, and
- wherein L is A(L=A) or wherein L is of formula V
- and wherein W allows of cleaving the peptide moiety under weakly acidic conditions of 0.1% to 30% trifluoroacetic acid.
Dependencies are inferred from the claim text.