Short chain PEGylation of amino acid monomers glutamine, lysine and peptides formed thereby

Patent metadata

Field	Value
Publication number	US-10280194-B2
Application number	US-201515317208-A
Country	US
Kind code	B2
Filing date	Jun 8, 2015
Priority date	Jun 9, 2014
Publication date	May 7, 2019
Grant date	May 7, 2019

Abstract

Novel synthesized amino acids of glutamine and lysine that are directly PEGylated with small, monodisperse PEGs, and a novel process for creating novel amino acid monomers using PEGylation. These amino acids are readily incorporated into peptides for a range of different applications.

First claim

The invention claimed is:

1. An amino acid selected from the group consisting of:

wherein:
R 1 is selected from the group consisting of hydrogen, allyloxycarbonyl, t-butoxycarbonyl, benzyloxycarbonyl, fluorenylmethyloxycarbonyl, trityl, 4-methyltrityl, 4-methoxytrityl, 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl, and 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutyl;
R 2 is selected from the group consisting of hydrogen, hydroxyl, alkyl of 1 to 5 carbons, benzyl, succinamide, 4-nitrophenyl, and pentafluorophenyl;
R 3 and R 4 are independently selected from the group consisting of hydrogen, —(CH 2 CH 2 O)mR 6 and —(CH 2 ) p R 7 ;
R 5 is —(CH 2 CH 2 O) m R 6 ;
R 6 is selected from the group consisting of alkyl of 1 to 5 carbons and —(CH 2 ) p R 7 ;
m is an integer of 1 to 25;
p is an integer of 1 to 10; and
R 7 is selected from the group consisting of allyloxycarbonylamine, t-butoxycarbonylamine, benzyloxycarbonylamine, fluorenylmethyloxycarbonylamine, tritylamine, 4-methyltritylamine, 4-methoxytritylamine, 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethylamine, 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutylamine, hydroxyl, methoxy, tert-butoxy, benzyloxy, trityloxy, cholesteroloxy, acetate, carboxylic acid, methyl ester, tert-butyl ester, benzyl ester, azide, alkyne, biotin, biotinamide, cholesterol and fluorescent molecules;
and
X 1 is a counterion to balance the charge; and

wherein:
R 8 is selected from the group consisting of hydrogen, allyloxycarbonyl, t-butoxycarbonyl, benzyloxycarbonyl, fluorenylmethyloxycarbonyl, trityl, 4-methyltrityl, 4-methoxytrityl, 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl, and 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutyl;
R 9 is selected from the group consisting of hydrogen, hydroxyl, alkyl of 1 to 5 carbons and benzyl;
R 10 represents a chemical bond or is selected from the group consisting of —NH(CH 2 ) q —where q is an integer of 1 to 10 and —NHCH 2 CH 2 (OCH 2 CH 2 ) r — where r is an integer of 1 to 10;
R 11 and R 12 represent a lone pair of electrons otherwise R 11 and R 12 are independently selected from the group consisting of a hydrogen, —(CH 2 CH 2 O) s R 14 and —(CH 2 ) t R 15 ;
R 13 is —(CH 2 CH 2 O) s R 14 ;
R 14 is selected from the group consisting of alkyl of 1 to 5 carbons and —(CH 2 ) t R 15 ;
s is an integer of 1 to 25;
t is an integer of 1 to 10; and
R 15 is selected from the group consisting of allyloxycarbonylamine, t-butoxycarbonylamine, benzyloxycarbonylamine, fluorenylmethyloxycarbonylamine, tritylamine, 4-methyltritylamine, 4-methoxytritylamine, 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethylamine, 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutylamine, hydroxyl, methoxy, tert-butoxy, benzyloxy, trityloxy, cholesteroloxy, acetate, carboxylic acid, methyl ester, tert-butyl ester, benzyl ester, azide, alkyne, biotin, biotinamide, cholesterol and fluorescent molecules;
and
X 2 is a counterion to balance the charge if necessary.

2. The amino acid of claim 1 wherein R 1 is selected from the group consisting of hydrogen, t-butoxycarbonyl, fluorenylmethyloxycarbonyl and benzyloxycarbonyl.

3. The amino acid of claim 1 wherein R 2 is selected from the group consisting of hydrogen, hydroxyl, methyl, ethyl, t-butyl, benzyl, succinamide, 4-nitrophenyl, and pentafluorophenyl.

4. The amino acid of claim 3 wherein R 2 is hydroxyl.

5. The amino acid of claim 1 wherein R 6 is methyl.

6. The amino acid of claim 1 wherein at least two of R 3 , R 4 and R 5 are the same.

7. The amino acid of claim 6 wherein at least two of R 3 , R 4 and R 5 are —(CH 2 CH 2 O) m CH 3 wherein m is 1 to 3.

8. The amino acid of claim 1 wherein at least two of R 3 , R 4 and R 5 are —(CH 2 CH 2 O) m R 6 wherein m is 1 to 4.

9. The amino acid of claim 8 wherein R 6 is —(CH 2 ) t R 7 wherein t is an integer of 2 to 4 and R 7 is selected from the group consisting of allyloxycarbonylamine, t-butoxycarbonylamine, tritylamine, 4-methyltritylamine, 4-methoxytritylamine, 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethylamine, 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutylamine, azide, alkyne, biotinamide and fluorescent molecules.

10. The amino acid of claim 1 wherein R 8 is selected from the group consisting of hydrogen, t-butoxycarbonyl, fluorenylmethyloxycarbonyl and benzyloxycarbonyl.

11. The amino acid of claim 1 wherein R 9 is selected from the group consisting of hydrogen, methyl, ethyl, t-butyl, and benzyl.

12. The amino acid of claim 1 wherein R 9 is hydroxyl.

13. The amino acid of claim 1 wherein R 10 represents a chemical bond.

14. The amino acid of claim 1 wherein R 10 is —NHCH 2 CH 2 —.

15. The amino acid of claim 1 wherein at least two of R 11 , R 12 and R 13 are the same.

16. The amino acid of claim 15 wherein at least two of R 11 , R 12 and R 13 are —(CH 2 CH 2 O) s CH 3 wherein s is 1 to 3.

17. The amino acid of claim 1 wherein at least two of R 11 , R 12 and R 13 are —(CH 2 CH 2 O) s R 14 wherein s is 1 to 4.

18. The amino acid of claim 17 wherein R 14 is —(CH 2 ) t R 15 wherein t is an integer of 2 to 4 and R 15 is selected from the group consisting of allyloxycarbonylamine, t-butoxycarbonylamine, tritylamine, 4-methyltritylamine, 4-methoxytritylamine, 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethylamine, 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutylamine, azide, alkyne, biotinamide and fluorescent molecules.

19. A method for forming a peptide comprising:
providing a first PEGylated amino acid selected from the group consisting of:

wherein:
R 1 is selected from the group consisting of hydrogen, allyloxycarbonyl, t-butoxycarbonyl, benzyloxycarbonyl, fluorenylmethyloxycarbonyl, trityl, 4-methyltrityl, 4-methoxytrityl, 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl, and 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutyl;
R 2 is selected from the group consisting of hydrogen, hydroxyl, alkyl of 1 to 5 carbons, benzyl, succinamide, 4-nitrophenyl, and pentafluorophenyl;
R 3 and R 4 are independently selected from the group consisting of hydrogen, —(CH 2 CH 2 O) m R 6 and —(CH 2 ) p R 7 ;
R 5 is —(CH 2 CH 2 O) m R 6 ;
R 6 is selected from the group consisting of alkyl of 1 to 5 carbons and —(CH 2 ) p R 7 ;
m is an integer of 1 to 25;
p is an integer of 1 to 10; and
R 7 is selected from the group consisting of allyloxycarbonylamine, t-butoxycarbonylamine, benzyloxycarbonylamine, fluorenylmethyloxycarbonylamine, tritylamine, 4-methyltritylamine, 4-methoxytritylamine, 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethylamine, 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutylamine, hydroxyl, methoxy, tert-butoxy, benzyloxy, trityloxy, cholesteroloxy, acetate, carboxylic acid, methyl ester, tert-butyl ester, benzyl ester, azide, alkyne, biotin, biotinamide, cholesterol and fluorescent molecules;
and
X 1 is a counterion to balance the charge;

Assignees

• Univ Clemson

Inventors

• Wetzler Modi
• Hamilton Paris Lamont

Classifications

• C07C271/22

  to carbon atoms of hydrocarbon radicals substituted by carboxyl groups · CPC title

• C07K2/00

  Peptides of undefined number of amino acids; Derivatives thereof · CPC title

• C07C237/12

  having the nitrogen atom of at least one of the carboxamide groups bound to an acyclic carbon atom of a hydrocarbon radical substituted by carboxyl groups (peptides C07K) · CPC title

• C07K1/066 Primary

  for omega-amido functions · CPC title

• C07C229/26

  having more than one amino group bound to the carbon skeleton, e.g. lysine · CPC title