Method for inverse solid phase synthesis of peptides

Abstract

The present invention provides a process for preparing a peptide of formula (I): Sub-[L]-[NH-A-C(O)] n+m —OH   (I) comprising: (a) reacting an immobilized compound of formula (II): Sub-(L)-[NH-A-C(O)] n —OH   (II) with an amino acid ester or peptide derivative of formula (III): H—[NH-A-C(O)] m —O(tBu)   (III) in the presence of a coupling agent to yield a peptide compound of general formula (IV): Sub-[L]-[NH-A-C(O)] n+m —O(tBu);   (IV) (b) removing the tBu (t-butyl) group to produce a solid-support bound carboxylic acid or peptide derivative of general formula (I); wherein n is a positive integer, e.g., 1-10, preferably 1-5; m is a positive integer.

1 - 41 . (canceled) 
     
     
         42 . An immobilized peptide derivative of
 a) formula (II) or (VI):
   Sub-[L]-[NH-A-C(O)] n —OH  (II) 
   Sub-[L]-[NH-A-C(O)] n —O(tBu)  (VI); 
   wherein n is a positive integer, Sub is a solid support, L is a linker to the amino acid ester or peptide derivative, each NH-A-C(O) independently is the residue of an optionally-protected amino acid or the structure —NH-A represents a heterocyclic group; the immobilized peptide derivative of formula (II) or (VI) being prepared by   reacting a functionalized solid support of general formula Sub[L]X with a compound of formula H—[NH-A-C(O)] n —O(tBu), wherein X is a group that links L to NH 2 -A by elimination of XH to provide the immobilized peptide derivative of formula (VI), and then   removing the t-Bu group to yield the immobilized peptide derivative of formula (II); or   b) formula (I) or (IV):
   Sub-[L]-[NH-A-C(O)] n+m —OH  (I) 
   Sub-[L]-[NH-A-C(O)] n+m —O(tBu)  (IV); 
   wherein m is a positive integer;   the immobilized peptide derivative of formula (I) or (IV) being prepared by reacting the immobilized peptide derivative of formula (II):
   Sub-[L]-[NH-A-C(O)] n —OH  (II) 
   
       with an amino acid ester derivative of formula (III):
   H—[NH-A-C(O)] m —O(tBu)  (III) 
 
       in the presence of a coupling agent to provide an immobilized peptide derivative of formula (IV):
   Sub-[L]-[NH-A-C(O)] n+m —O(tBu)  (IV); 
 
       and then,
 removing the t-Bu (t-butyl) group to provide an immobilized peptide derivative of formula (I); or 
 c) formula (VIII) or (XX):
   Sub-[L]-[NH-A-C(O)] n+m(x+1) —OH  (VIII) 
   Sub-[L]-[NH-A-CO] n+m(x+1) —O(tBu)  (XX); 
 
 wherein x is an integer greater than or equal to 0; 
 the immobilized peptide derivative of formula (VII) or (XX) being prepared by carrying out, x times, the steps of reacting the immobilized peptide derivative of formula (I)
   Sub-[L]-[NH-A-C(O)] n+m —OH  (I) 
 
 
       with an amino acid ester derivative of formula (III)
   H—[NH-A-C(O)] m —O(tBu)  (III) 
 
       to provide the immobilized peptide derivative of formula (XX), and then
 removing the t-Bu group to yield an immobilized peptide derivative of formula (VIII). 
 
     
     
         43 . The immobilized peptide derivative of  claim 42  wherein n=1. 
     
     
         44 . The immobilized peptide derivative of  claim 42  wherein m=1. 
     
     
         45 . The immobilized peptide derivative of  claim 42  wherein x=1. 
     
     
         46 . The immobilized peptide derivative of  claim 42  wherein [L]X is OC(O)(CH 2 ) 2 CH(NHZ)CO 2 H, wherein Z is a carbobenzoxy group. 
     
     
         47 . The immobilized peptide derivative of  claim 42  wherein [L]X is HNC(O)(CH 2 ) 2 CH(NHZ)CO 2 H, wherein Z is a carbobenzoxy group. 
     
     
         48 . The immobilized peptide derivative of  claim 42  wherein each NH-A-C(O) is individually the optionally sidechain-protected residue of an α-amino acid. 
     
     
         49 . The immobilized peptide derivative of  claim 42  wherein each NH-A-C(O) is individually the optionally sidechain-protected residue of a naturally occurring L-amino acid. 
     
     
         50 . The immobilized peptide derivative of  claim 42  wherein the coupling agent is HATU. 
     
     
         51 . The immobilized peptide derivative of  claim 42  wherein a step of removing a t-Bu group is carried out using TFA. 
     
     
         52 . The immobilized peptide derivative of  claim 42  wherein Sub is a polymer comprising free NH 2  or OH groups. 
     
     
         53 . The immobilized peptide derivative of  claim 52  wherein the polymer comprises polystyrene. 
     
     
         54 . An immobilized peptide derivative of formula (XII):
   Sub-[L]-[NH-A-C(O)] n+m —NH(alkyl)C(O)CF 3   (XII)   
       prepared by reacting the immobilized peptide derivative of formula (I) of  claim 42  with
   H 2 N(alkyl)CH(OH)CF 3   (XV) 
 
       in the presence of a coupling agent to yield the corresponding amide of the immobilized peptide derivative of formula (I), and then oxidizing the CH(OH) moiety to yield the immobilized peptide derivative of general formula (XII). 
     
     
         55 . An immobilized peptide derivative of formula (XVI):
   Sub-[L]-[NH-A-C(O)] n+m(x+1) —NH(alkyl)C(O)CF 3   (XVI)   
       prepared by reacting the immobilized peptide derivative of formula (VIII) of  claim 42  with compound (XV):
   H2N(alkyl)CH(OH)CF3  (XV) 
 
       in the presence of a coupling agent to yield the corresponding amide of the immobilized peptide derivative of formula (VIII) and then oxidizing the CH(OH) moiety to yield the immobilized peptide derivative of formula (XVI). 
     
     
         56 . An immobilized peptide derivative of formula (XIII):
   Sub-[L]-[NH-A-C(O)] n+m —NH(alkyl)CHO  (XIII).   
       prepared by reacting the immobilized peptide derivative of formula (I) of  claim 42  with
   H 2 N(alkyl)OH  (XVII) 
 
       in the presence of a coupling agent to yield the hydroxyalkylamide of (I), and then oxidizing the terminal hydroxy group to yield the immobilized peptide derivative of formula (XIII). 
     
     
         57 . An immobilized peptide derivative of formula (XVIII):
   Sub-[L]-[NH-A-C(O)] n+m(x+1) —NH(alkyl)CHO  (XVIII)   
       prepared by reacting the immobilized peptide derivative of formula (VIII) of  claim 42  with
   H 2 N(alkyl)OH  (XVII) 
 
       in the presence of a coupling agent to yield the hydroxyalkylamide of (VIII), and then oxidizing the terminal hydroxyl group to yield the immobilized peptide derivative of formula (XVIII). 
     
     
         58 . An immobilized peptide derivative of formula (XXI) comprising the immobilized peptide derivative of formula (XIII) of  claim 56  wherein the terminal aldehyde group is protected as an acetal or a thioacetal. 
     
     
         59 . An immobilized peptide derivative of formula (XXII) comprising the immobilized peptide derivative of formula (XVII) of  claim 57  wherein the terminal aldehyde group is protected as an acetal or a thioacetal. 
     
     
         60 . An immobilized peptide derivative of formula (XIV):
   Sub-[L]-[NH-A-C(O)] n+m(x+1) —NH(A)B(OR) 2   (XIV),   
       wherein R is (C 1 -C 6 )alkyl, phenyl, or the residue of a diol and wherein NH(A)B(OR) 2  is the residue of an O-protected aminoboronic acid, prepared by reacting an immobilized peptide derivative of formula (VEII) of  claim 42  with a boroamine in the presence of a coupling agent to yield the immobilized peptide derivative of formula (XIV). 
     
     
         61 . An immobilized peptide derivative of formula (XIX):
   Sub-[L]-[NH-A-C(O)] n+m —NH(A)B(OR) 2   (XIX),   
       wherein R is (C 1 -C 6 )alkyl, phenyl, or the residue of a diol and wherein NH(A)B(OR) 2  is the residue of an O-protected aminoboronic acid, prepared by reacting an immobilized peptide derivative of formula (I) of  claim 42  with a boroamine in the presence of a coupling agent to yield the compound of formula (XIX). 
     
     
         62 . An immobilized peptide derivative of formula (I) or (IV):
   Sub-[L]-[NH-A-C(O)] n+m —OH  (I)     Sub-[L]-[NH-A-C(O)] n+m —O(tBu)  (IV)   wherein n is a positive integer, m is a positive integer, Sub is a solid support, L is a linker to the amino acid ester or peptide derivative, each NH-A-C(O) independently is the residue of an optionally-protected amino acid or the structure —NH-A represents a heterocyclic group.   
     
     
         63 . An immobilized peptide derivative of formula (II) or (VI):
   Sub-[L]-[NH-A-C(O)] n —OH  (II)     Sub-[L]-[NH-A-C(O)] n —O(tBu)  (VI)   wherein n is a positive integer, Sub is a solid support, L is a linker to the amino acid ester or peptide derivative, each NH-A-C(O) independently is the residue of an optionally-protected amino acid or the structure —NH-A represents a heterocyclic group.   
     
     
         64 . An immobilized peptide derivative of formula (VII) or (XX)
   Sub-[L]-[NH-A-C(O)] n+m(x+1) —OH  (VIII)     Sub-[L]-[NH-A-CO] n+m(x+1) —O(tBu)  (XX)   wherein n is a positive integer, m is a positive integer, x is an integer greater than or equal to 0, Sub is a solid support, L is a linker to the amino acid ester or peptide derivative, each NH-A-C(O) independently is the residue of an optionally-protected amino acid or the structure —NH-A represents a heterocyclic group.