Accelerants for the Modification of Non-Natural Amino Acids and Non-Natural Amino Acid Polypeptides

• US 20090137736A1
• Filed: 11/08/2006
• Published: 05/28/2009
• Est. Priority Date: 11/08/2005
• Status: Abandoned Application

• Current Assignee: Ambrx, Inc. (Johnson & Johnson)
• Original Assignee: Ambrx, Inc. (Johnson & Johnson)
• Inventors: Miao, Zhenwei, Tian, Feng
• Application Number:
  US12/092,265
• Publication Number:
  US 20090137736A1
• Time in Patent Office: Days
• US Class Current: 525/54.100
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 1/1077 by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids
• CPC Class Codes: C07K 14/61 Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: Y02P 20/55 Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Abstract

Disclosed herein are accelerants for the formation of oxime-containing compounds from the reaction of a carbonyl-containing compound and a hydroxylamine-containing compound. The oxime-containing compound, the carbonyl-containing compound and the hydroxylamine-containing compound can each be a non-natural amino acid or a non-natural amino acid polypeptide. Also disclosed is the use of such accelerants to form oxime-containing compounds, the resulting oxime-containing compounds, and reaction mixtures containing such accelerants.

Claims

1. A reaction mixture comprising a compound comprising an aromatic ketone moiety, a compound comprising a hydroxylamine moiety, and an accelerant selecting from the group consisting of bifunctional aromatic amines, oxoamine derivatives, and compounds having the following structures:

wherein R_x, R_y and R_z are selected from the group consisting of;

L_x-H, L_x-alkyl, L_x-aryl, L_x-heteroaryl, L_x-alkenyl, L_x-alkynyl, L_x-alkoxy, and L_x-alkylamine, where L_x is a bond, C(═

O), C(═

NH), C(═

NH)—

NH, SO, and SO₂.

2. The reaction mixture of claim 1, wherein the compound comprising an aromatic ketone moiety is an amino acid or a polypeptide.

3. The reaction mixture of claim 1, wherein the compound comprising an aromatic ketone moiety has the structure:

wherein;

R is alkyl, substituted alkyl, cycloalkyl, or substituted cycloalkyl;

R₁ is H, an amino protecting group, resin, amino acid, polypeptide, or polynucleotide; and

R₂ is OH, an ester protecting group, resin, amino acid, polypeptide, or polynucleotide;

wherein each R_a is independently selected from the group consisting of H, halogen, alkyl, substituted alkyl, —

N(R′

)₂, —

C(O)_kR′

where k is 1, 2, or 3, —

C(O)N(R′

)₂, —

OR′

, and —

S(O)_kR′

,

4. The reaction mixture of claim 1, wherein the compound comprising a hydroxylamine moiety further comprises a polymer moiety.

5. The reaction mixture of claim 1, wherein the compound comprising a hydroxylamine moiety has the structure:

wherein;

each L is a linker independently selected from the group consisting of alkylene, substituted alkylene, alkenylene, substituted alkenylene, —

O—

, —

O-(alkylene or substituted alkylene)-, —

S—

, —

S-(alkylene or substituted alkylene)-, —

S(O)_k—

where k is 1, 2, or 3, —

S(O)_k(alkylene or substituted alkylene)-, —

C(O)—

, —

C(O)-(alkylene or substituted alkylene)-, —

C(S)—

, —

C(S)-(alkylene or substituted alkylene)-, —

N(R′

)—

, —

NR′

-(alkylene or substituted alkylene)-, —

C(O)N(R′

)—

, —

CON(R′

)-(alkylene or substituted alkylene)-, -(alkylene or substituted alkylene)NR′

C(O)O-(alkylene or substituted alkylene)-, —

O—

CON(R′

)-(alkylene or substituted alkylene)-, —

CSN(R′

)—

, —

CSN(R′

)-(alkylene or substituted alkylene)-, —

N(R′

)CO-(alkylene or substituted alkylene)-, —

N(R′

)C(O)O—

, —

N(R′

)C(O)O-(alkylene or substituted alkylene)-, —

S(O)_kN(R′

)—

, —

N(R′

)C(O)N(R′

)—

, —

N(R′

)C(O)N(R′

)-(alkylene or substituted alkylene)-, —

N(R′

)C(S)N(R′

)—

, —

N(R′

)S(O)_kN(R′

)—

, —

N(R′

)—

N—

, —

C(R′

)═

N—

, —

C(R′

)═

N—

N(R′

)—

, —

C(R′

)═

N—

N═

, —

C(R′

)₂—

N═

N—

, and —

C(R′

)₂—

N(R′

)—

N(R′

)—

, where each R′

is independently H, alkyl, or substituted alkyl.

6. The reaction mixture of claim 1, wherein the accelerant is a bifunctional aromatic amine.

7. The reaction mixture of claim 1, wherein the accelerant is an oxoamine derivative.

8. A method for derivatizing an amino acid of Formula (III), the method comprising contacting the amino acid with a reagent of Formula (XXVII) in the presence of an accelerant, wherein Formula (III) corresponds to:

wherein;

R is alkyl, substituted alkyl, cycloalkyl, or substituted cycloalkyl;

R₁ is H, an amino protecting group, resin, amino acid, polypeptide, or polynucleotide; and

R₂ is OH, an ester protecting group, resin, amino acid, polypeptide, or polynucleotide;

wherein each R_a is independently selected from the group consisting of H, halogen, alkyl, substituted alkyl, —

N(R′

)₂, —

C(O)_kR′

where k is 1, 2, or 3, —

C(O)N(R′

)₂, —

OR′

, and —

S(O)_kR′

,

wherein Formula (XXVII) corresponds to;

wherein;

each L is a linker independently selected from the group consisting of alkylene, substituted alkylene, alkenylene, substituted alkenylene, —

O—

, —

O-(alkylene or substituted alkylene)-, —

S—

, —

S-(alkylene or substituted alkylene)-, —

S(O)_k—

where k is 1, 2, or 3, —

S(O)_k(alkylene or substituted alkylene)-, —

C(O)—

, —

C(O)-(alkylene or substituted alkylene)-, —

C(S)—

, —

C(S)-(alkylene or substituted alkylene)-, —

N(R′

)—

, —

NR′

-(alkylene or substituted alkylene)-, —

C(O)N(R′

)—

, —

CON(R′

)-(alkylene or substituted alkylene)-, -(alkylene or substituted alkylene)NR′

C(O)O-(alkylene or substituted alkylene)-, —

O—

CON(R′

)-(alkylene or substituted alkylene)-, —

CSN(R′

)—

, —

CSN(R′

)-(alkylene or substituted alkylene)-, —

N(R′

)CO-(alkylene or substituted alkylene)-, —

N(R′

)C(O)O—

, —

N(R′

)C(O)O-(alkylene or substituted alkylene)-, —

S(O)_kN(R′

)—

, —

N(R′

)C(O)N(R′

)—

, —

N(R′

)C(O)N(R′

)-(alkylene or substituted alkylene)-, —

N(R′

)C(S)N(R′

)—

, —

N(R′

)S(O)_kN(R′

)—

, —

N(R′

)—

N═

, —

C(R′

)═

N—

, —

C(R′

)═

N—

N(R′

)—

, —

C(R′

)═

N—

N═

, —

C(R′

)₂—

N═

N—

, and —

C(R′

)₂—

N(R′

)—

N(R′

)—

, where each R′

is independently H, alkyl, or substituted alkyl; and

wherein the accelerant is selected from the group consisting of bifunctional aromatic amines, oxoamine derivatives, and compounds having the following structures;

wherein R_x, R_y and R_z are selected from the group consisting of;

L_x-H, L_x-alkyl, L_x-aryl, L_x-heteroaryl, L_x-alkenyl, L_x-alkynyl, L_x-alkoxy, and L_x-alkylamine, where L_x is a bond, C(═

O), C(═

NH), C(═

NH)—

NH and SO, SO₂.

9. The method of claim 8, wherein R is alkyl.

10. The method of claim 8, wherein R is CH₃.

11. The method of claim 8, wherein the accelerant has the structure:

H₂N—

NH—

C(O)—

R_b, wherein R_b is alkyl, substituted alkyl, NH—

NH₂, H, and alkoxy.

12. The method of claim 11, wherein R_b is alkyl or alkoxy.

13. The method of claim 8, wherein the accelerant is selected from the group consisting of the compounds identified as 6, 7, 8, 10, and 20 of FIG. 5.

14. The method of claim 8, wherein the accelerant is a bifunctional aromatic amine.

15. The method of claim 14, wherein the bifunctional aromatic amine is selected from the group consisting of:

Bifunctional aromatic amines;

16. The method of claim 8, wherein the accelerant is an oxoamine derivative.

17. The method of claim 16, wherein the oxoamine is selected from the group consisting of:

Oxoamine derivatives;

18. The method of claim 8, wherein the molecular weight of the PEG group is between about 1,000 Da and about 40,000 Da.

19. The method of claim 8, wherein the amino acid is contacted with the reagent of Formula (XXVII) in aqueous solution at about room temperature.

20. The method of claim 8, wherein the amino acid is contacted with the reagent of Formula (XXVII) in aqueous solution at a pH between about 4 to about 10.

21. The method of claim 8, wherein the molar ratio of amino acid to the reagent of Formula (XXVII) is selected from the group of about 1:

2;

1;

1;

1.5;

1;

1.5;

2;

2;

1;

1;

1.5;

2;

1.5; and

1.5 to 2.

22. The method of claim 8, wherein the amino acid of Formula (III) has been incorporated site-specifically during the in vivo translation of a polypeptide.

23. The method of claim 8, wherein the derivatized amino acid comprises at least one oxime containing amino acid having the structure of Formula (XI-A):

wherein;

R is H, alkyl, substituted alkyl, cycloalkyl, or substituted cycloalkyl;

R₁ is H, an amino protecting group, resin, amino acid, polypeptide, or polynucleotide; and

R₂ is OH, an ester protecting group, resin, amino acid, polypeptide, or polynucleotide;

wherein each R_a is independently selected from the group consisting of H, halogen, alkyl, substituted alkyl, —

N(R′

)₂, —

C(O)_kR′

where k is 1, 2, or 3, —

C(O)N(R′

)₂, —

OR′

, and —

S(O)_kR1,

R₅ is L-X, where

X is a PEG; and

L is optional, and when present is a linker selected from the group consisting of alkylene, substituted alkylene, alkenylene, substituted alkenylene, —

O—

, —

O-(alkylene or substituted alkylene)-, —

S—

, —

S-(alkylene or substituted alkylene)-, —

S(O)_k—

where k is 1, 2, or 3, —

S(O)_k(alkylene or substituted alkylene)-, —

C(O)—

, —

C(O)-(alkylene or substituted alkylene)-, —

C(S)—

, —

C(S)-(alkylene or substituted alkylene)-, —

N(R′

)—

, —

NR′

-(alkylene or substituted alkylene)-, —

C(O)N(R′

)—

, —

CON(R′

)-(alkylene or substituted alkylene)-, —

CSN(R′

)—

, —

CSN(R′

)-(alkylene or substituted alkylene)-, —

N(R′

)CO-(alkylene or substituted alkylene)-, —

N(R′

)C(O)O—

, —

S(O)_kN(R′

)—

, —

N(R′

)C(O)N(R′

)—

, —

N(R′

)C(S)N(R′

)—

, —

N(R′

)S(O)_kN(R′

)—

, —

N(R′

)—

N═

, —

C(R′

)═

N—

, —

C(R′

)═

N—

N(R′

)—

, —

C(R′

)═

N—

N═

, —

C(R′

)₂—

N═

N—

, and —

C(R′

)₂—

N(R′

)—

N(R′

)—

, where each R′

is independently H, alkyl, or substituted alkyl.

24. The derivatized amino acid of claim 23.

25. The derivatized amino acid of claim 24, wherein the amino acid is incorporated into a therapeutic protein that is a member of the growth hormone supergene family.

26. The derivatized amino acid of claim 25, wherein the therapeutic protein is human growth hormone.