Solid support for Fmoc-solid phase synthesis of peptides
View Patent ↗The present invention provides compositions and processes for the solid phase synthesis of polypeptides. In particular, the present invention provides solid supports and processes for preparing solid supports for the synthesis of polypeptides.
1. A compound comprising Formula (I):
wherein:
R 2 and R 4 are hydrocarbyl;
R 3 is selected from the group consisting of hydrogen, a protecting group, an amino acid residue, and a peptide;
R 5 is a solid support comprising at least one polymer; and
R 6 , R 7 , R 8 , and R 9 are independently selected from the group consisting of hydrogen, hydrocarbyl, and substituted hydrocarbyl.
2. The compound of claim 1 , wherein R 3 is selected from the group consisting of Fmoc and a peptide; and R 5 is polystyrene cross-linked with divinylbenzene.
3. The compound of claim 2 , wherein the peptide has a C-terminus secondary amine.
4. The compound of claim 1 , comprising:
wherein:
R 3 is selected from the group consisting of hydrogen, a protecting group, an amino acid residue, and a peptide;
R 5 is a solid support comprising at least one polymer;
m is an integer from 1 to 8; and
n is an integer from 1 to 8.
5. The compound of claim 4 , wherein R 3 is selected from the group consisting of Fmoc and a peptide; and R 5 is polystyrene cross-linked with divinylbenzene.
6. The compound of claim 5 , wherein the peptide has a C-terminus secondary amine.
7. The compound of claim 1 , comprising:
wherein:
R 3 is selected from the group consisting of hydrogen, a protecting group, an amino acid residue, and a peptide; and
R 5 is a solid support comprising at least one polymer.
8. The compound of claim 7 , wherein R 3 is selected from the group consisting of Fmoc and a peptide; and R 5 is polystyrene cross-linked with divinylbenzene.
9. The compound of claim 8 , wherein the peptide has a C-terminus secondary amine.
10. The compound of claim 8 , wherein the peptide is selected from the group consisting of Leuprolide, Deslorelin, Buserelin, Alarelin, Fertirelin, and Histrelin.
11. A process for making a solid support, the process comprising:
(a) combining a compound comprising formula (1) with a compound comprising formula (2) in the presence of a base to produce a compound comprising formula (3), wherein the compound comprising formula (1) has the following structure:
wherein:
R 4 is hydrocarbyl; and
R 5 is a solid support comprising at least one polymer;
the compound comprising formula (2) has the following structure:
wherein:
R 6 , R 7 , R 8 , and R 9 are independently selected from the group consisting of hydrogen, hydrocarbyl, and substituted hydrocarbyl; and
the compound comprising formula (3) has the following structure:
wherein:
R 4 is hydrocarbyl;
R 5 is a solid support comprising at least one polymer; and
R 6 , R 7 , R 8 , and R 9 are independently selected from the group consisting of hydrogen, hydrocarbyl, and substituted hydrocarbyl;
(b) contacting the compound comprising formula (3) with a reducing agent and a compound comprising R 2 NH 2 , wherein R 2 is hydrocarbyl, to produce a solid support comprising formula (I);
wherein:
R 2 and R 4 are hydrocarbyl;
R 5 is a solid support comprising at least one polymer; and
R 6 , R 7 , R 8 , and R 9 are independently selected from the group consisting of hydrogen, hydrocarbyl, and substituted hydrocarbyl.
12. The process of claim 11 , wherein the base is selected from the group consisting of sodium methoxide, potassium carbonate, lithium carbonate, sodium carbonate, magnesium carbonate, calcium carbonate, and strontium carbonate; and the amount of compound 1 to the amount of compound 2 to the amount of base is a molar ratio ranging from about 1:1:1 to about 1:5:5.
13. The process of claim 11 , wherein the reducing agent is selected from the group consisting of sodium borohydride, sodium cyanoborohydride, diisobutylaluminium hydride, lithium aluminium hydride, and titanium isopropoxide sodium borohydride; and the amount of compound 3 to the amount of R 2 NH 2 to the amount of reducing agent is a molar ratio ranging from about 1:1:1 to about 1:10:20.
14. The process of claim 11 , wherein the base is potassium carbonate; the reducing agent is sodium borohydride; the amount of compound 1 to the amount of compound 2 to the amount of potassium carbonate is a molar ratio ranging from about 1:1:1 to about 1:5:5; and the amount of compound 3 to the amount of R 2 NH 2 to the amount of sodium borohydride is a molar ratio ranging from about 1:1:1 to about 1:10:20.
15. The process of claim 11 , wherein:
the compound comprising formula (1) has the following structure:
the compound comprising formula (2) has the following structure:
the compound comprising formula (3) has the following structure:
and the compound comprising formula (I) has the following structure:
wherein R 5 is solid support comprising at least one polymer.
16. The process of claim 15 , wherein R 5 is polystyrene cross-linked with divinylbenzene; the base is potassium carbonate; the R 2 NH 2 is ethylamine; the reducing agent is sodium borohydride; the amount of compound 1 to the amount of compound 2 to the amount about of potassium carbonate is a molar ratio ranging from about 1:1:1 to about 1:5:5; and the amount of compound 3 to the amount of ethylamine to the amount of sodium borohydride is a molar ratio ranging from about 1:1:1 to about 1:10:20.
17. A method for synthesizing a polypeptide, the method comprising:
(a) activating the carboxyl group of an amino acid that has its amine protected by a Fmoc group, and its side chain protected by an acid labile group;
(b) coupling the activated amino acid to the amino group of a solid support comprising Formula (I):
wherein:
R 2 and R 4 are hydrocarbyl;
R 5 is a solid support comprising at least one polymer; and
R 6 , R 7 , R 8 , and R 9 are independently selected from the group consisting of hydrogen, hydrocarbyl, and substituted hydrocarbyl;
(c) treatment of the solid support with a base to deprotect the amine group of the amino acid protected with Fmoc; and
(d) repeating steps (a) to (c) until the target polypeptide is synthesized.
18. The method of claim 17 , wherein the polypeptide is selected from the group consisting of Leuprolide, Deslorelin, Buserelin, Alarelin, Fertirelin, and Histrelin.
19. The method of claim 17 , further comprising cleaving the target polypeptide from the solid support by contacting the solid support of step (d) with a weak acid and a scavenger selected from the group consisting of phenol, water, 1,2-ethanedithiol, and triisopropylsilane.
20. The method of claim 17 , wherein the solid support has the following structure:
wherein R 5 is polystyrene cross-linked with divinylbenzene.