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US 2014/0235789 A1Published application
Solid phase peptide synthesis via side chain attachment
AssigneeChemical & Biopharmaceutical Laboratories of Patras S.A.
InventorKleomenis K. BARLOS
Life of this patent
Estimating the term…
Abstract
The present application discloses peptides and peptaibols of high purity may be obtained by solid phase peptide synthesis using as the starting resin hydroxy amino acids, hydroxy amino acid amides, hydroxy amino alcohols or small peptides containing hydroxy amino acids attached to polymers through their side chain.
Claims25
A resin conjugate of the formula I or II, wherein:
- Hya is the residue of a hydroxy amino acid;
- Pr1 is H, or an amino protecting group which is orthogonal to the resin and to other protecting groups;
- A is a hydroxyl group or a acid sensitive hydroxyl protecting group selected from tBu, Trt, Clt, or NR1R2 wherein R1 and R2 are H or C1-10 alkyl, or peptide ester or peptide amide or a peptaibol containing 0-30 amino acids;
- X, Y, Z and V are each independently a substituent on the ortho, meta or para positions and is selected from H, Cl, F, C1-10 alkyl or C1-10 alkoxy;
- and
- P is an insoluble solid support or an insoluble linker-resin conjugate suitable for the solid phase synthesis of peptides
- or
- a resin conjugate of the formula III-VI, wherein:
- Pr1 is H or an amino protecting group, which is orthogonal to the resin and to other protecting groups;
- Pr2 is H or a hydroxyl protecting group which is orthogonal to the resin;
- R3 and R4 are each independently H or C1-10 alkyl;
- X, Y, Z and V are each independently on the ortho, meta or para positions selected from H, Cl, F, C1-10 alkyl or C1-10 alkoxy;
- and
- P is an insoluble solid support or an insoluble linker-resin conjugate suitable for the solid phase synthesis of peptides.
11 more elements
(canceled)
A process for the preparation of monoalkylated Fmoc-amino di-alcohols, the process comprising the reaction of the Fmoc-di-amino alcohol with alkylhalide in the presence of a base such as a tertiary amine base in an organic solvent, such as dichloromethane;
- and wherein alkyl is a triarylmethyl selected from trityl, 4-methyltrityl, 4-methoxytrityl and 2-chlorotrityl.
A peptide of the formula E-D-2-Nal-Cys(A)-Tyr(C)-D-Trp(F)-Lys(E)-Val-Cys(A)-Thr(Resin)-NH2 wherein:
- D—designates the chirality of the amino acid that follows as a D-amino acid;
- each A is independently a thiol protecting group selected from Trt, Mmt, Acm or StBu;
- C is a hydroxy protecting group selected from Clt, Trt or tBu;
- F is H or Boc;
- E is an amino protecting group selected from Mtt, Mmt, Trt or Boc;
- and
- Resin is H or an acid labile resin suitable for solid-phase peptide synthesis.
4 more elements
A peptide of the formula E-Phe-Val-Asn(A)-Gln(A)-His(A)-Leu-Cys(B)-Gly-Ser(C)-His(A)-Leu-Val-Glu(C)-Ala-Leu-Tyr(C)-Leu-Val-Cys(A)-Gly-Glu(C)-Arg(D)-Gly-Phe-Phe-Tyr(C)-Thr(C)-Pro-Lys(E)-Thr(Resin)-O —C wherein:
- A is H or a carboxamido or imidazol protecting group selected from Trt, Mtt or Mmt;
- B is a Cys protecting group selected from Mmt, Trt, Acm or StBu;
- C is a hydroxy, carboxyl or phenoxy protecting group selected from tBu, Trt or Clt;
- D is a guanidine protecting group selected from Pbf or Pmc;
- E is H or an amino protecting group selected from Fmoc, Boc, Trt, Nps, Mtt, Mmt or Clt;
- and
- Resin is H or an acid labile resin suitable for solid-phase peptide synthesis
- or
- a peptide of the formula E-Ser(C)-His(A)-Leu-Val-Glu(C)-Ala-Leu-Tyr(C)-Leu-Val-Cys(B)-Gly-Glu(C)-Arg(D)-Gly-Phe-Phe-Tyr(C)-Thr(C)-Pro-Lys(E)-Thr(Resin)-O— wherein:
- A is a carboxamido or imidazol protecting group selected from Trt, Mtt or Mmt;
- B is a Cys protecting group selected from Mmt, Trt, Acm or StBu;
- Each C is independently a hydroxy, carboxyl or phenoxy protecting group selected from tBu, Trt or Clt;
- D is H or a guanidine protecting group selected from Pbf or Pmc;
- E is H or an amino protecting group selected from Fmoc, Boc, Trt, Nps, Mtt, Mmt or Clt;
- and
- Resin is H or an acid labile resin suitable for solid-phase peptide synthesis
- or
- a peptide of the formula E-Cys(B)-Ser(C)-Asn(A)-Leu-Ser(C)-Thr(C)-Cys(B)-Val-Leu-Gly-Lys(E)-Leu-Ser(C)-Gln(A)-Glu(C)-Leu-His(A)-Lys(E)-Leu-Gln(A)-Thr(C)-Tyr(C)-Pro-Arg(D)-Thr(C)-Asn(A)-Thr(C)-Gly-Ser(C)-Gly-Thr(Resin)-Pro-NH2, wherein:
- each A is independently a carboxamido or imidazol protecting group selected from Trt, Mtt or Mmt;
- each B is independently a Cys protecting group selected from Mmt, Trt, Acm or StBu;
- each C is independently a hydroxyl, carboxy or phenoxy protecting group selected from of tBu, Trt or Clt;
- D is a guanidine protecting group selected from Pbf or Pmc;
- each E is independently H or an amino protecting group selected from Fmoc, Boc, Trt, Nps, Mtt, Mmt or Clt;
- and
- Resin is H or an acid labile resin suitable for solid-phase peptide synthesis
- or
- a partially protected peptide of the formula E-His(A)-Gly-Glu(C)-Gly-Thr(C)-Phe-Thr(C)-Ser(C)-Asp(C)—X-Lys(E)-Gln(A)-Met-Glu(C)-Glu(C)-Glu(C)-Ala-Val-Arg-(D)-Leu-Phe-Ile-Glu(C)-Trp(F)-Leu-Lys(E)-Asn(A)-Gly-Gly-Pro-Ser(C)-Ser(C)-Gly-Ala-Pro-Pro-Pro-Ser(Resin)-NH2 wherein:
- X is Leu-Ser(tBu) or Leu-ΨSer;
- each A is independently H or a carboxamido or imidazol protecting group selected from Trt, Mtt or Mmt;
- each C is independently a hydroxy, carboxyl or phenoxy protecting group selected from tBu, Trt or Clt;
- D is H or a guanidine protecting group elected from Pbf or Pmc;
- each E is independently H or an amino protecting group selected from Fmoc, Boc, Trt, Nps, Mtt, Mmt or Clt;
- F is H or Boc;
- and
- Resin is an acid labile resin suitable for solid-phase peptide synthesis
- or
- a partially protected peptide of the formula E-Lys(E)-Cys(B)-Asn(A)-Thr(C)—Y-Cys(B)—Y-Gln(A)-Arg(D)-Leu-Ala-Asn(A)-Phe-Leu-Val-His(A)-X-Asn(A)-Asn(A)-Phe-Gly-Pro-Ile-Leu-Pro-Pro-Thr(C)-Asn(A)-Val-Gly-Ser(C)-Asn(A)-Thr(C)-Tyr(Resin)-NH2 wherein X=Ser(tBu)-Ser(tBu) or Ser(tBu)-ΨSer, wherein:
- Y is Ala-Thr(tBu) or Ala-ΨThr;
- each A is independently H, a carboxamido or imidazol protecting group selected from Trt, Mtt or Mmt;
- each B is independently a Cys protecting group selected from Mmt, Trt, Acm or StBu;
- each C is independently hydroxyl, carboxy or phenoxy protecting group selected from tBu, Trt or Clt;
- D is H or a guanidine protecting group selected from Pbf or Pmc;
- E is H or an amino protecting group selected from Fmoc, Boc, Trt, Nps, Mtt, Mmt or Clt;
- and
- Resin is H or an acid labile resin suitable for solid-phase peptide synthesis
- or
- a protected or partially protected peptide with the sequence Z-Asn(A)-Phe-Leu-Val-His(A)-X-Asn(A)-Asn(A)-Phe-Gly-Pro-Ile-Leu-Pro-Pro-Thr(C)-Asn(A)-Val-Gly-Ser(C)-Asn(A)-Thr(C)-Tyr(Resin)-NH2 in particular the protected or partially protected 10-38, 11-38, 12-38 and 14-38 partially protected pramlintide fragments wherein:
- Z is H or E-Gln(A)-Arg(D)-Leu-Ala, E-Arg(D)-Leu-Ala, E-Leu-Ala or E-Ala;
- X is Ser(tBu)-Ser(tBu) or Ser(tBu)-ΨSer;
- A is H or a carboxamido or imidazol protecting group selected from Trt, Mtt or Mmt;
- C is a hydroxyl, carboxy or phenoxy protecting group selected from tBu, Trt or Clt;
- D is H or a guanidine protecting group selected from Pbf or Pmc;
- E is H or an amino protecting group selected from Fmoc, Boc, Trt, Nps, Mtt, Mmt and Clt;
- and
- Resin is H or an acid labile resin suitable for solid-phase peptide synthesis
- or
- a protected or partially protected peptide of the formula E-D-Phe-Pro-Arg(D)-Pro-Gly-Gly-Gly-Gly-Asn(A)-Gly-Asp(C)-Phe-Glu(C)-Glu(C)-Ile-Pro-Glu(C)-Glu(C)-Tyr(Resin)-Leu-O—C, wherein:
- A is H or a carboxamido or imidazol protecting group selected from Trt, Mtt or Mmt;
- each C is independently a hydroxyl or carboxyl protecting group selected from tBu, Trt or Clt;
- D is H or a guanidine protecting group selected from Pbf or Pmc;
- E is H or an amino protecting group selected from Fmoc, Boc, Trt, Nps, Mtt, Mmt or Clt;
- and
- Resin is H or an acid labile resin suitable for solid-phase peptide synthesis
- or
- a protected or partially protected peptide with the formula E-Ser(C)-Tyr(C)-Ser(C)-Met-Glu(C)-His(A)-Phe-Arg(D)-Trp(F)-Gly-Lys(E)-Pro-Val-Gly-Lys(E)-Lys(E)-Arg(D)-Arg(D)-Pro-Val-Lys(E)-Val-Tyr(Resin)-Pro-O—C, wherein:
- A is H or a carboxamido or imidazol protecting group selected from Trt, Mtt or Mmt;
- each C is independently a hydroxy, carboxyl or phenoxy protecting group selected from tBu, Trt or Clt;
- each D is independently H or a guanidine protecting group selected from Pbf or Pmc;
- each E is independently H or an amino protecting group selected from Fmoc, Boc, Trt, Nps, Mtt, Mmt or Clt;
- F is H or Boc and
- Resin is H or an acid labile resin suitable for solid-phase peptide synthesis.
68 more elements
13-20. (canceled)
A partially protected peptide of the formula:
- wherein:
- A is H or an amino protecting group selected from Fmoc, Boc, Trt, Nps, Mtt or Mmt;
- D—designates the chirality of the amino acid that follows as a D-amino acid;
- B is a thiol protecting group selected from Trt, Mmt, Acm or StBu;
- C is H or Boc;
- E is a hydroxy protecting group selected from Clt, Trt or tBu;
- and
- Resin is H or an acid labile resin suitable for solid-phase peptide synthesis.
5 more elements
25-26. (canceled)
A partially protected peptide Y-Glu(C)-Glu(C)-Ala-Val-Arg(D)-Leu-Phe-Ile-Glu(C)-Trp(F)-Leu-Lys(E)-Asn(A)-Gly-Gly-Pro-Ser(C)-Ser(C)-Gly-Ala-Pro-Pro-Pro-Ser(Resin)-NH2 that is the exenatide fragments 12-39, 13-39, 14-39, wherein:
- Y is H, E-Glu(C), E-Met-Glu(C), E-Gln(A)-Met-Glu(C) or E-Lys(E′)-Gln(A)-Met-Glu(C);
- wherein:
- each A is independently H or a carboxamido protecting group selected from Trt, Mtt and Mmt;
- E and E′ are each independently H or an amino protecting group selected from Fmoc, Boc, Trt, Nps, Mtt, Mmt or Clt;
- F is H or Boc;
- each C is independently a hydroxy or carboxyl protecting group selected from tBu, Trt or Clt;
- D is H or a guanidine protecting group selected from Pbf or Pmc;
- and
- Resin is H or an acid labile resin suitable for solid-phase peptide synthesis
- or
- a partially protected peptides that is E-His(A)-Gly-Glu(C)-Gly-Thr(C)-Phe-Thr(C)-Ser(C)-Asp(C)-X—Y in particular sequences corresponding to the exenatide fragments 1-11 1-13, 1-14 and 1-15, wherein:
- X is Leu-Ser(tBu) or Leu-ΨSer;
- E is an amino protecting group selected from Fmoc, Boc, Trt, Nps, Mtt, Mmt and Clt;
- A is H, a carboxamido or imidazol protecting group selected from Trt, Mtt or Mmt;
- each C is independently a hydroxy or carboxyl protecting group selected from tBu, Trt or Clt;
- and
- Y is OZ, Lys(E)-Gln(A)-OZ, Lys(E)-Gln(A)-Met-OZ, Lys(E)-Gln(A)-Met-Glu(C)—OZ, wherein each Z is independently H or a group which activate electrophilically carboxyl groups selected from Bt, Su, Pfp, Tcp or Pnp.
14 more elements
(canceled)
30-32. (canceled)
A process for the preparation of a partially protected and oxidized pramlintide fragments, comprising:
- assembling protected peptides on an acid sensitive resin, such as a 2-CTC-resin;
- cleaving the protected peptide from the resin;
- and
- oxidizing the resin-bound peptides by contacting the resin-bound peptides with mild acid, solvent comprises a 2-200 molar excess of iodine over the fragment.
1 more element
35-42. (canceled)
Dependencies are inferred from the claim text.