Compound or salt thereof and preparation method and application of same

Abstract

The invention relates to a compound or a salt thereof, a method for preparation thereof, and use thereof, wherein the compound has the structure of Formula (1):the substituents in Formula (1) are as defined in the specification. The compound of Formula (1) or a salt thereof can be attached to a solid-phase resin, on which solid-phase synthesis may be performed.

1 . A compound represented by Formula (1) or a salt thereof: 
       
         
           
           
               
               
           
         
         in Formula (1), R 1  is hydrogen or an amino-protecting group; 
         R 2 , together with the N atom R 2  is attached to as an amino group, form an amino acid, or an amino acid with a protecting group, or a peptide chain formed by them; 
         X 1  and X 2  are each independently O or S; 
         R 3  is H or a substituent containing an active group that can react with an attaching functional group on a solid-phase resin; 
         R 4  is H or an alkyl group. 
       
     
     
         2 . The compound according to  claim 1  or a salt thereof, wherein the amino-protecting group includes Fmoc, Boc, Alloc, Dde, ivDde, Trt, Mtt or Mmt. 
     
     
         3 . The compound according to  claim 1  or a salt thereof, wherein the amino acid or amino acid with a protecting group is an α-amino acid or an α-amino acid with a protecting group. 
     
     
         4 . The compound according to  claim 1  or a salt thereof, wherein R 2  is 
       
         
           
           
               
               
           
         
       
       wherein P is an amino-protecting group or a hydroxyl-protecting group, and multiple Ps can be the same or different. 
     
     
         5 . The compound according to  claim 4  or a salt thereof, wherein R 2  is 
       
         
           
           
               
               
           
         
       
     
     
         6 . The compound according to  claim 5  or a salt thereof, wherein R 2  is 
       
         
           
           
               
               
           
         
       
     
     
         7 . The compound according to  claim 1  or a salt thereof, wherein R 3  is a substituent containing carboxyl, hydroxyl and/or NH 2 . 
     
     
         8 . The compound according to  claim 1  or a salt thereof, wherein R 3  is H or —(CH 2 ) m COOH, m is an integer from 1 to 10, and any one or more CH 2  in the (CH 2 ) m  may have a substituent. 
     
     
         9 . The compound according to  claim 1  or a salt thereof, wherein R 3  and X 1  together form —OH, —O(CH 2 ) m COOH, —SH or —O(CH 2 ) m COOH. 
     
     
         10 . The compound according to  claim 1  or a salt thereof, wherein both X 1  and X 2  are O. 
     
     
         11 . The compound according to  claim 1  or a salt thereof, wherein Formula (1) has the following structures: 
       
         
           
           
               
               
           
         
         in Formulae (VI), (IX), (X) and (XII), R 1 -R 4 , X 1 , X 2 , and m are as defined in any one of  claims 1 - 10 ; 
         R 5  is selected from the group consisting of —CH 2 COOH, —CH 2 CH 2 COOH, 
       
       
         
           
           
               
               
           
         
       
       —(CH 2 ) 4 NH 2 , 
       
         
           
           
               
               
           
         
       
       —CH 2 OH, —CH(OH)CH 3 , —CH 2 CONH 2 , —CH 2 CH 2 CONH 2 , —CH 2 SH, H, —CH 3 , —CH(CH 3 ) 2 , —CH 2 CH(CH 3 ) 2 , —CH(CH 3 )CH 2 CH 3 , —(CH 2 ) 2 SCH 3 , 
       
         
           
           
               
               
           
         
       
       when R 5  contains a carboxyl, hydroxyl, —NH 2 , —NH—, sulfydryl, or guanidine group, it may be protected by a protecting group;
 R 6  is a carboxyl-protecting group. 
 
     
     
         12 . The compound according to  claim 1  or a salt thereof, wherein Formula (1) has the following structures: 
       
         
           
           
               
               
           
         
       
     
     
         13 . (canceled) 
     
     
         14 . A solid-phase resin carrying the compound of Formula (1) according to  claim 1  or a salt thereof, wherein the solid-phase resin carrying the compound or a salt thereof has the following structure: 
       
         
           
           
               
               
           
         
         in Formula (1)-resin,   represents the body of the solid-phase resin, G is a linking structure formed by a reaction between a group in R 3  in Formula (1) according to  claim 1  and a group on the side chain of the solid-phase resin, R′ 3  is the residual structure after the reaction of R 3 , Z is the residual structure after the reaction of the side chain of the solid-phase resin, and the definitions of the other substituents are the same as those in Formula (1); preferably, R′ 3 -G-Z is R′ 3 —CONH—Z, or R′ 3 —COO—Z; 
         alternatively, Z is directly connected to X 1  without R′ 3 -G. 
       
     
     
         15 . The solid-phase resin according to  claim 14 , wherein Formula (1)-resin has the following structures: 
       
         
           
           
               
               
           
         
         preferably, Formula (1)-resin has the following structures: 
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         preferably, in Formula (i)-resin to Formula (iv)-resin, and Formula (ix)-resin to Formula (x)-resin, the resin is selected from an AM resin, an MBHA resin, a Sieber resin or a Rink resin, preferably an AM resin or an MBHA resin; 
         preferably, in Formula (ix)-resin to Formula (x)-resin, the resin is a CTC resin. 
       
     
     
         16 . Use of the compound of Formula (1) according to  claim 1  or a salt thereof or a Formula (1) resin, in the preparation of a target polypeptide or a salt thereof, wherein the C terminal of the target polypeptide contains the —NHR 2  group from Formula (1); or alternatively, this —NHR 2  group may be transformed into a target structure or form a ring with the rest of the polypeptide chain. 
     
     
         17 . The use according to  claim 16 , comprising the following steps:
 attaching the compound of Formula (1) to a starting solid-phase resin to obtain Formula (1)-resin, or directly providing Formula (1)-resin;   
       
         
           
           
               
               
           
         
       
       in Formula (1)-resin,   represents the body of the solid-phase resin, G is a linking structure formed by a reaction between a group in R 3  in Formula (1) according to  claim 1  any one of  claims 1 - 12  and a group on the side chain of the solid-phase resin, R′ 3  is the residual structure after the reaction of R 3 , Z is the residual structure after the reaction of the side chain of the solid-phase resin, and the definitions of the other substituents are the same as those in Formula (1); preferably, R′ 3 -G-Z is R′ 3 -CONH—Z, or R′ 3 -COO—Z; 
       alternatively, Z is directly connected to X 1  without R′ 3 -G, 
       with the proviso that: when R 1  is an amino-protecting group, it is removed to expose the NH group; when R 1  is hydrogen, there is no need to remove it;
 preparing a peptide-resin with the peptide chain fully protected by stepwise solid-phase coupling on the exposed NH group; 
 cleaving the fully protected peptide chain off the peptide-resin to obtain, directly or after removing the protecting groups, the target polypeptide or a salt thereof; 
 optionally, before cleaving the fully protected peptide chain off the peptide-resin, using the fully protected peptide chain as a starting peptide to carryout cyclization to form a cyclic peptide; preferably, the reaction site of cyclization in the fully protected peptide chain is in R 2 ; 
 wherein the Fmoc-protecting group attached to the peptide on the resin is removed before each coupling step; 
 when the target polypeptide is PMX-53 or a salt thereof, which is a cyclic peptide, the NR 2  in the Formula (1)-resin is 
 
       
         
           
           
               
               
           
         
       
       wherein P 1  and P 2  are protecting groups, preferably, the NR 2  is 
       
         
           
           
               
               
           
         
       
       Formula (1)-resin may be Formula (VI)-resin 
       
         
           
           
               
               
           
         
       
       Formula (VI)-resin may be Formula (ix)-resin 
       
         
           
           
               
               
           
         
       
       preferably, the resin in Formula (ix)-resin is a CTC resin;
 more preferably, when the target polypeptide is PMX-53 or a salt thereof, the peptide-resin with the uncyclized PMX-53 fully protected is prepared b successive solid-phase coupling of the amino acids Fmoc-Trp(Boc)-OH, Fmoc-D-Cha-OH, Fmoc-Pro-OH, Fmoc-Orn(Dde)-OH, and Fmoc-Phe-OH as reaction reagents to the exposed NH 2  group, followed by acetylation of the amino group of the last amino acid coupled, thereby obtaining the following peptide-resin with the uncyclized PMX-53 fully protected; 
 Ac-Phe-Orn(Dde)-Pro-D-Cha-Trp(Boc)-Formula (1)-resin; preferably, Ac-Phe-Orn(Dde)-Pro-D-Cha-Trp(Boc)-Formula (VI)-resin; more preferably, Ac-Phe-Orn(Dde)-Pro-D-Cha-Trp(Boc)-Formula (ix)-resin; even more preferably, Ac-Phe-Orn(Dde)-Pro-D-Cha-Trp(Boc)-Formula (ix)-CTC resin; 
 wherein the Fmoc-protecting group attached to the peptide on the resin is removed before each coupling step; 
 then the All protecting group and the Dde protecting group are removed, followed by a cyclization reaction to obtain the peptide resin with PMX-53 fully protected: Ac-Phe-c(Orn-Pro-D-Cha-Trp(Boc)-Arg(pbf))-resin, wherein the resin is preferably a CTC resin; 
 finally, the protecting groups are removed by cleavage to obtain PMX-53 or a salt thereof; 
 when the target polypeptide is Liraglutide or a salt thereof, its C terminal being —NCH 2 COOH, the NR 2  in Formula (1)-resin is NCH 2 COOP, in which P is a protecting group, preferably tert-butyl; Formula (1)-resin may be Formula (VI)-resin; Formula (VD-resin may be Formula (x)-resin defined in  claim 15 ; 
 
       
         
           
           
               
               
           
         
       
       preferably, the resin in Formula (x)-resin may be an AM resin, an MBHA resin, a Sieber resin, or a Rink resin, among which the first two resins are preferred;
 further preferably, when the target polypeptide is Liraglutide or a salt thereof, the peptide-resin with Liraglutide fully protected is prepared by successive solid-phase coupling of the following amino acids as reaction reagents to the exposed NH 2  group: moc-Arg(pbf)-OH, Fmoc-Gly-OH, Fmoc-Arg(pbf)-OH, Fmoc-Val-OH, Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc-Ala-OH, Fmoc-Ile-OH, Fmoc-Phe-OH, Fmoc-Glu(Ot-Bu)-OH, Fmoc-Lys(N-s-(Palm-Glu-Ot-Bu)-OH, Fmoc-Ala-OH, Fmoc-Ala-OH, Fmoc-Gln(Trt)-OH, Fmoc-Gly-OH, Fmoc-Glu(Ot-Bu)-OH, Fmoc-Leu-OH, Fmoc-Tyr(t-Bu)-OH, Fmoc-Ser(t-Bu)-OH, Fmoc-Ser(t-Bu)-OH, Fmoc-Val-OH, Fmoc-Asp(Ot-Bu)-OH, Fmoc-Ser(t-Bu)-OH, Fmoc-Thr(t-Bu)-OH, Fmoc-Thr(t-Bu)-OH, Fmoc-HmbGly-OH, Fmoc-Glu(Ot-Bu)-OH, Fmoc-Ala-OH, Boc-His(Trt)-OH; thereby obtaining the following peptide-resin with Liraglutide fully protected; 
 H-His(Trt)-Ala-Glu(Ot-Bu)-HmbGly-Thr(t-Bu)-Phe-Thr(t-Bu)-Ser(t-Bu)-Asp(Ot-Bu)-Val-Ser(t-Bu)-Ser(t-Bu)-Tyr(t-Bu)-Leu-Glu(Ot-Bu)-Gly-Gln(Trt)-Ala-Ala-Lys(N-ε-(Palm-Glu-Ot-Bu))-Glu(Ot-Bu)-Phe-Ile-Ala-Trp(Boc)-Leu-Val-Arg(pbf)-Gly-Arg(pbf)-Formula (1)-resin; preferably, H-His(Trt)-Ala-Glu(Ot-Bu)-HmbGly-Thr(t-Bu)-Phe-Thr(t-Bu)-Ser(t-Bu)-Asp(Ot-Bu)-Val-Ser(t-Bu)-Ser(t-Bu)-Tyr(t-Bu)-Leu-Glu(Ot-Bu)-Gly-Gln(Trt)-Ala-Ala-Lys(N-ε-(Palm-Glu-Ot-Bu))-Glu(Ot-Bu)-Phe-Ile-Ala-Trp(Boc)-Leu-Val-Arg(pbf)-Gly-Arg(pbf)-Formula (VI)-resin; more preferably, H-His(Trt)-Ala-Glu(Ot-Bu)-HmbGly-Thr(t-Bu)-Phe-Thr(t-Bu)-Ser(t-Bu)-Asp(Ot-Bu)-Val-Ser(t-Bu)-Ser(t-Bu)-Tyr(t-Bu)-Leu-Glu(Ot-Bu)-Gly-Gln(Trt)-Ala-Ala-Lys(N-ε-(Palm-Glu-Ot-Bu))-Glu(Ot-Bu)-Phe-Ile-Ala-Trp(Boc)-Leu-Val-Arg(pbf)-Gly-Arg(pbf)-Formula (x)-resin; further preferably, H-His(Trt)-Ala-Glu(Ot-Bu)-HmbGly-Thr(t-Bu)-Phe-Thr(t-Bu)-Ser(t-Bu)-Asp(Ot-Bu)-Val-Ser(t-Bu)-Ser(t-Bu)-Tyr(t-Bu)-Leu-Glu(Ot-Bu)-Gly-Gln(Trt)-Ala-Ala-Lys(N-ε-(Palm-Glu-Ot-Bu))-Glu(Ot-Bu)-Phe-Ile-Ala-Trp(Boc)-Leu-Val-Arg(pbf)-Gly-Arg(pbf)-Formula (x)-AM resin; 
 wherein the Fmoc protecting group attached to the peptide on the resin is removed before each coupling step.