Chemical reagents for formation of disulfide bonds in peptides
AssigneeThe Regents of the University of California
InventorsDallas L. Rabenstein, Tiesheng Shi
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Abstract
This invention pertains to the discovery of a class of reagents that effectively form intramolecular disulfide bonds in peptides. Intermolecular disulfide linkage formation is low or essentially non-existent. In addition, preferred reagents of this invention are relatively mild and do not oxidize “vulnerable” residues in the subject peptide(s). In addition the reagents and reaction products are safe and essentially non-toxic. One particularly preferred reagent is [Pt(en)2Cl2]2+ where en is ethylenediamine.
Claims46
A method of forming an intramolecular disulfide bond in a peptide, said method comprising:
- contacting a peptide comprising at least two sulfhydryl (SH) groups with a compound having the formula:
- wherein R1 and R2 are independently selected from the group consisting of Cl—, Br—, I—, OH—, and RCO2— where R is an alkyl or substituted alkyl;
- Pt is Pt(IV);
- X1, X2, X3, and X4 are ligands independently selected from the group consisting of a nitrogen ligand, an oxygen ligand, a phosphorous ligand, and a sulfur ligand;
- i, k, m, and p are independently 0 or 1;
- L1, L2, L3, and L4 when present, are independently selected linkers.
A solution comprising:
- a peptide comprising at least two sulfhydryl groups;
- and
- a compound having the formula:
- wherein R1 and R2 are independently selected from the group consisting of Cl—, Br—, I—, OH—, and RCO2— where R is an alkyl or substituted alkyl;
- Pt is Pt(IV);
- X1, X2, X3, and X4 are ligands independently selected from the group consisting of a nitrogen ligand, an oxygen ligand, a phosphorous ligand, and a sulfur ligand;
- i, k, m, and p are independently 0 or 1;
- L1, L2, L3, and L4 when present, are independently selected linkers.
A method of chemically synthesizing a peptide comprising a disulfide linkage, said method comprising:
- chemically coupling a plurality of amino acids to form a peptide comprising at least two sulfhydryl (SH) groups;
- and
- contacting said peptide with a reagent having the formula:
- wherein R1 and R2 are independently selected from the group consisting of Cl—, Br—, I—, OH—, and RCO2— where R is an alkyl of substituted alkyl;
- Pt is Pt(IV);
- X1, X2, X3, and X4 are ligands independently selected from the group consisting of a nitrogen ligand, an oxygen ligand, a phosphorous ligand, and a sulfur ligand;
- i, k, m, and p are independently 0 or 1;
- L1, L2, L3, and L4 when present, are independently selected linkers.
A kit for forming an intramolecular disulfide bond in a peptide, said kit comprising:
- a container containing a reagent having the formula:
- wherein R1 and R2 are independently selected from the group consisting of Cl—, Br—, I—, OH—, and RCO2— where R is an alkyl or substituted alkyl;
- Pt is Pt(IV);
- X1, X2, X3, and X4 are ligands independently selected from the group consisting of a nitrogen ligand, an oxygen ligand, a phosphorous ligand, and a sulfur ligand;
- i, k, m, and p are independently 0 or 1;
- L1, L2, L3, and L4 when present, are independently selected linkers.
Dependencies are inferred from the claim text.