Process for the Production of Bivalirudin

Abstract

The present invention relates to a process for the production of bivalirudin, a 20-mer peptide of formula H- D -Phe 1 -Pro-Arg-Pro-Gly 5 -Gly-Gly-Gly-Asn-Gly 10 -Asp-Phe-Glu-Glu-Ile 15 -Pro-Glu-Glu-Tyr-Leu 20 -OH   (I) via a convergent five-fragment synthesis, and to several peptide intermediates thereof.

1 - 18 . (canceled) 
     
     
         19 . A peptide selected from the group consisting of
 (i) an optionally side chain-protected peptide of formula
   P1- D -Phe 1 -Pro-Arg-Pro-OH (SEQ ID NO 4)   (V),
 
 wherein P1 is one of Bpoc, Ddz, Fmoc, Adc, Aoc, Dpp, Msc or Pht; 
   (ii) an optionally side chain-protected peptide of formula
   H-Gly 5 -Gly-Gly-Gly-Asn-Gly 10 -OP2 (SEQ ID NO 5)   (VI),
 
 wherein P2 is a protecting group being removable by catalytic hydrogenation and being orthogonal to the optional side chain protecting group(s); 
   (iii) an optionally side chain-protected peptide of formula
   P1- D -Phe 1 -Pro-Arg-Pro-Gly 5 -Gly-Gly-Gly-Asn-Gly 10 -OP2 (SEQ ID NO 2)   (VII),
 
 wherein P1 and P2 are as defined above; 
   (iv) a side chain-protected peptide of formula
   P1- D -Phe 1 -Pro-Arg-Pro-Gly 5 -Gly-Gly-Gly-Asn-Gly 10 -OH (SEQ ID NO 2)   (IIa),
 
 wherein P1 is as defined above, except for Boc- D -Phe 1 -Pro-Arg(Pbf)-Pro-Gly 5 -Gly-Gly-Gly-Asn(Trt)-Gly 10 -OH; 
   (v) a side chain-unprotected peptide of formula
   P1- D -Phe 1 -Pro-Arg-Pro-Gly 5 -Gly-Gly-Gly-Asn-Gly 10 -OH (SEQ ID NO 2)   (IIb),
 
 wherein P1 is as defined above; 
   (vi) a side chain-protected peptide of formula
   H-Glu-Glu-Ile 15 -Pro-Glu-Glu-Tyr-Leu 20 -OP3 (SEQ ID NO 8)   (X),
 
 wherein P3 is a protecting group being removable by catalytic hydrogenation; 
   (vii) a side chain-protected peptide of formula
   P4-Phe-Glu-Glu-Ile 15 -Pro-Glu-Glu-Tyr-Leu 20 -OP3 (SEQ ID NO 9)   (XII),
 
 wherein P3 is as defined above; and P4 is a protecting group being orthogonal to the side chain protecting group(s) of the peptide of formula X and to P3; or P4 is hydrogen; 
   (viii) a side chain-protected peptide of formula
   P5-Asp-Phe-Glu-Glu-Ile 15 -Pro-Glu-Glu-Tyr-Leu 20 -OP3 (SEQ ID NO 3)   (XIV),
 
 wherein P5 is a protecting group being orthogonal to the side chain protecting group(s) of the peptide/amino acid of formula XII and XIII and to P3; and P3 is as defined above; except for Fmoc-Asp(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ile 15 -Pro-Glu(OtBu)-Glu(OtBu)-Tyr(tBu)-Leu 20 -OtBu; 
   (ix) a side chain-protected peptide of formula
   H-Asp-Phe-Glu-Glu-Ile 15 -Pro-Glu-Glu-Tyr-Leu 20 -OP3 (SEQ ID NO 3)   (III),
 
 wherein P3 is as defined above; except for H-Asp(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ile 15 -Pro-Glu(OtBu)-Glu(OtBu)-Tyr(tBu)-Leu 20 -OtBu; and 
   (x) a side chain-protected peptide of formula
   P1- D -Phe 1 -Pro-Arg-Pro-Gly 5 -Gly-Gly-Gly-Asn-Gly 10 -Asp-Phe-Glu-Glu--Ile 15 -Pro-Glu-Glu-Tyr-Leu 20 -OP3 (SEQ ID NO 1)   (IV),
 
 wherein P1 and P3 are as defined above; except for Boc- D -Phe 1 -Pro-Arg(Pbf)-Pro-Gly 5 -Gly-Gly-Gly-Asn(Trt)-Gly 10 -Asp(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ile 15 -Pro-Glu(OtBu)-Glu(OtBu)-Tyr(tBu)-Leu 20 -OtBu. 
   
     
     
         20 . (canceled) 
     
     
         21 . The peptide of  claim 19 , wherein the optionally side chain-protected peptide of formula VI is H-Gly 5 -Gly-Gly-Gly-Asn-Gly 10 -OBzl (SEQ ID NO 5). 
     
     
         22 . The peptide of  claim 19 , wherein the optionally side chain-protected peptide of formula VII is
   Boc- D -Phe 1 -Pro-Arg-Pro-Gly 5 -Gly-Gly-Gly-Asn-Gly 10 -OBzl (SEQ ID NO 2).   
     
     
         23 . The peptide of  claim 19 , wherein the side chain-unprotected peptide of formula IIb is
   Boc- D -Phe 1 -Pro-Arg-Pro-Gly 5 -Gly-Gly-Gly-Asn-Gly 10 -OH (SEQ ID NO 2).   
     
     
         24 . The peptide of  claim 19 , wherein the side chain-protected peptide of formula X is
   H-Glu(OBzl)-Glu(OBzl)-Ile 15 -Pro-Glu(OBzl)-Glu(OBzl)-Tyr(Bzl)-Leu 20 -OBzl (SEQ ID NO 8).   
     
     
         25 . The peptide of  claim 19 , wherein the side chain-protected peptide of formula XII is
   Boc-Phe-Glu(OBzl)-Glu(OBzl)-Ile 15 -Pro-Glu(OBzl)-Glu(OBzl)-Tyr(Bzl)-Leu 20 -OBzl (SEQ ID NO 9),     or     H-Phe-Glu(OBzl)-Glu(OBzl)-Ile 15 -Pro-Glu(OBzl)-Glu(OBzl)-Tyr(Bzl)-Leu 20 -OBzl (SEQ ID NO 9).   
     
     
         26 . The peptide of  claim 19 , wherein the side chain-protected peptide of formula XIV is
   Boc-Asp(OBzl)-Phe-Glu(OBzl)-Glu(OBzl)-Ile 15 -Pro-Glu(OBzl)-Glu(OBzl)-Tyr(Bzl)-Leu 20 -OBzl (SEQ ID NO 3).   
     
     
         27 . The peptide of  claim 19 , wherein the side chain-protected peptide of formula III is
   H-Asp(OBzl)-Phe-Glu(OBzl)-Glu(OBzl)-Ile 15 -Pro-Glu(OBzl)-Glu(OBzl)-Tyr(Bzl)-Leu 20 -OBzl (SEQ ID NO 3).   
     
     
         28 . The peptide of  claim 19 , wherein the side chain-protected peptide of formula IV is
   Boc- D -Phe 1 -Pro-Arg-Pro-Gly 5 -Gly-Gly-Gly-Asn-Gly 10 -Asp(OBzl)-Phe-Glu(OBzl)-Glu(OBzl)-Ile 15 -Pro-Glu(OBzl)-Glu(OBzl)-Tyr(Bzl)-Leu 20 -OBzl (SEQ ID NO 1).   
     
     
         29 . Use of any of the peptides of  claim 19  as an intermediate in a synthesis of bivalirudin.