Method of peptide synthesis
AssigneeResearch Corporation Technologies, Inc.
InventorsLouis A. Carpino, Dumitru Ionescu
Life of this patent
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Abstract
The present invention relates to a process for forming an N-α-amino protected amino acid fluoride in situ by reacting an N-α-amino protected amino acid with an ionic fluoride salt in the presence of a peptide coupling agent. It is also directed to the use of the amino acid fluoride thus formed in peptide synthesis.
Claims98
An in situ process for preparing a N-∝-protected amino acid fluoride which comprises mixing a N-∝ protected amino acid or acylating derivative thereof with a fluorinating effective amount of an ionic fluoride salt in the presence of a coupling agent under conditions effective to form said protected amino acid fluoride, wherein the anion of the ionic fluoride salt has the formula:
- Ly (HF)2F⊖, TG1G2G3F2— or LF⊖;
- wherein
- z is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;
- y is 0 or 1;
- L is TG1G2G3G4;
- T is a group IV element selected from the group consisting of Si, Ge, Sn and Pb;
- and
- G1, G2, G3 and G4 are independently halogen, hydrogen,, alkyl, aryl, arylalkyl, cycloalkyl, or cycloalkylalkyl.
A process for forming a peptide between a N-∝-amino protected amino acid or acylating derivative thereof and a second amino acid with a free amino group or a peptide containing a free amino group, which process comprises:
- (a) mixing in situ said N-∝-amino protected amino acid with a fluorinating effective amount of an ionic fluoride salt in the presence of a coupling agent under conditions effective to form a N-∝-amino protected amino acid fluoride, and
- (b) reacting the product of (a) with said second amino acid or peptide under peptide forming conditions, wherein the anion of the ionic fluoride salt has the formula:
- Ly (HF)zF−, TG1G2G3F2−or LF⊖;
- wherein
- z is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;
- y is 0 or 1;
- L is TG1G2G3G4;
- T is a group IV element selected from the group consisting of Si, Ge, Sn and Pb;
- and
- G1, G2, G3 G4 are independently halogen, hydrogen, alkyl, aryl, arylalkyl, cycloalkyl, or cycloalkylalkyl.
In the improved process for forming an organic acid fluoride in situ, from an organic acid, the improvement comprising reacting said organic acid with a fluorinating effective amount of an ionic fluoride salt in the presence of a peptide coupling agent, under conditions effective to form said organic acid fluoride, wherein the anion of the ionic fluoride salt has the formula:
- Ly (HF)zF−, TG1G2G3F2− or LF⊖,
- wherein
- z is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;
- y is 0 or 1;
- L is TG1G2G3G4;
- T is a group IV element selected from the group consisting of Si, Ge, Sn and Pb;
- and
- G1, G2, G3 G4 are independently halogen, hydrogen, alkyl, aryl, arylalkyl, cycloalkyl, or cycloalkylalkyl.
In the synthesis of a peptide wherein a first N-∝-amino protected amino acid is covalently coupled to a solid phase peptide synthesis resin, the N-∝-amino protecting group is cleaved off and the resulting free amino group is reacted under amide forming conditions with a N-∝-amino protected amino acid fluoride, the resulting N-∝-amino protecting group is cleaved off and coupled to another protected amino acid fluoride and the cycle repeated until the desired peptide has been obtained, and then the peptide is cleaved from said resin, the improvement comprising preparing said amino acid fluoride in situ by reacting a N-∝-amino protected amino acid or acylating derivative thereof with a fluorinating effective amount of an ionic fluoride salt in the presence of a coupling agent, under conditions effective to form said protected amino acid fluoride, wherein the anion of the ionic fluoride salt has the formula:
- Ly(HF)2Fe⊖, LF⊖ or TG1G2G3F2−;
- wherein
- z is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;
- y is 0 or 1;
- L is TG1G2G3G4;
- G1, G2, G3 and G4 are independently halogen, hydrogen, alkyl, aryl, aryl alkyl, cycloalkyl or cycloalkylalkyl;
- and
- T is a Group IV element selected from the group consisting of Si, Ge, Sn, and Pb.
In a process for forming an amide from the reaction of an organic amine and a carboxylic acid fluoride, the improvement comprising forming the carboxylic acid fluoride in situ by reacting a carboxylic acid or acylating derivative thereof with a fluorinating effective amount of an ionic fluoride salt in the presence of a peptide coupling agent under conditions effective to form said carboxylic acid fluoride, which is then reacted with the organic amine to form said amide, wherein the anion of the fluoride salt has the formula:
- Ly(HF)zF⊖, TG1G2G3G4F2− or LF−;
- wherein
- z is 1-10;
- y is 0 or 1;
- L is TG1G2G3G4;
- T is a Group IV element selected from the group consisting of Si, Ge, Sn and Pb;
- and
- G1, G2, G3 and G4 are independently halogen, hydrogen alkyl, aryl, arylalkyl, cycloalkyl or cycloalkylalkyl.
A solution comprising a fluorinating effective amount of an ionic fluoride salt having an ionizable fluoride, a coupling agent and an N-∝-amino protected amino acid or acylating derivative thereof, wherein the anion of the ionic fluoride salt has the formula:
- Ly(HF)zF−, TG1G2G3F2− or LF⊖;
- wherein
- z is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;
- y is 0 or 1;
- L is TG1G2G3G4;
- and
- T is a Group IV element selected from the group consisting of Si, Ge, Sn and Pb;
- and
- G1, G2, G3 and G4 are independently hydrogen, alkyl, aryl, arylalkyl, cycloalkyl or cycloalkylalkyl, wherein the fluoride salt, coupling agent and Nocamino protected amino acid or acylating derivative are capable of forming a N-∝-amino acid fluoride in situ.
A kit useful for preparing amino acid fluorides in situ comprising an ionic fluoride salt and an N-∝-amino protected amino acid, the contents of said kit, when mixed with a coupling agent in solution, generates an amino acid fluoride in situ, the anion of the ionic fluoride salt having the formula:
- Ly(HF)2F−, TG1G2G3F2− or LF⊖;
- wherein
- z is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;
- y is 0 or 1;
- L is TG1G2G3G4;
- T is a Group IV element selected from the group consisting of Si, Ge, Sn and Pb;
- and
- G1, G2, G3 and G4 are independently halogen, hydrogen, alkyl, aryl, arylalkyl, cycloalkyl or cycloalkylalkyl.
A kit useful for preparing amino acid fluorides in situ comprising an ionic fluoride salt in one compartment and a coupling agent in a different compartment, the contents of said kit when mixed with an N-∝-amino-protected amino acid or acylating derivative in solution generates an N-∝-amino protected-amino acid fluoride in situ, the anion of the ionic fluoride salt having the formula:
- Ly(HF)2F−, TG1G2G3F2− or LF⊖;
- wherein
- z is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;
- v is 0 or 1 ;
- L is TG1G2G3G4;
- T is a Group IV element selected from the group consisting of Si, Ge, Sn and Pb;
- and
- G1, G2, G3 and G4 are independently halogen, hydrogen, alkyl, aryl, arylalkyl, cycloalkyl or cycloalkylalkyl.
A process for forming an amino acid fluoride in situ comprising reacting a fluorinating effective amount of an ionic fluoride salt with a mixed anhydride of a N-∝-amino acid, under conditions effective to form said amino acid fluoride, wherein the anion of the ionic fluoride salt has the formula:
- Ly(HF)zF−, TG1G2G3F2− or LF⊖;
- wherein
- z is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;
- y is 0 or 1;
- L is TG1G2G3G4;
- T is a Group IV element selected from the group consisting of Si, Ge, Sn and Pb;
- and
- G1, G2, G3 and G4 are independently halogen, hydrogen, alkyl, aryl, arylalkyl, cycloalkyl or cycloalkylalkyl.
A process for forming an amino acid fluoride in situ comprising reacting under fluorination conditions a fluorinating effective amount of an ionic fluoride salt with an active ester of an N-∝-amino protected amino acid, under conditions effective to form said amino acid fluoride, wherein the anion of the ionic fluoride salt has the formula:
- Ly(HF)zF−, TG1G2G3F2− or LF⊖;
- wherein
- z is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;
- y is 0 or 1,
- L is TG1G2G3G4;
- T is a Group IV element selected from the group consisting of Si, Ge, Sn and Pb;
- and
- G1, G2, G3 and G4 are independently halogen, hydrogen, alkyl, aryl, arylalkyl, cycloalkyl or cycloalkylalkyl.
Dependencies are inferred from the claim text.