Modified amino acids

• US 9,682,934 B2
• Filed: 08/30/2013
• Issued: 06/20/2017
• Est. Priority Date: 08/31/2012
• Status: Active Grant

• Current Assignee: Sutro Biopharma, Inc.
• Original Assignee: Sutro Biopharma, Inc.
• Inventors: Yang, Wenjin, Stafford, Ryan, Thanos, Christopher D.
• Primary Examiner(s): Jarrell, Noble
• Assistant Examiner(s): Michelson, Ben S
• Application Number:
  US14/015,893
• Publication Number:
  US 20140066598A1
• Time in Patent Office: 1,390 Days
• Field of Search: None
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: C07C 247/04 being saturated
• CPC Class Codes: C07C 271/20 to carbon atoms of hydrocarbon radicals substituted by nitrogen atoms not being part of nitro or nitroso groups
• CPC Class Codes: C07C 275/14 being further substituted by nitrogen atoms not being part of nitro or nitroso groups
• CPC Class Codes: C07D 213/55 Acids; Esters
• CPC Class Codes: C07K 14/00 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: C07K 14/535 Granulocyte CSF; Granulocyte-macrophage CSF
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002

Abstract

Provided herein are modified amino acids comprising an azido group, polypeptides, antibodies and conjugates comprising the modified amino acids, and methods of producing the polypeptides, antibodies and conjugates comprising the modified amino acids. The polypeptides, antibodies and conjugates are useful in methods of treatment and prevention, methods of detection and methods of diagnosis.

Claims

1. A polypeptide which is a polymer of amino acid residues where one or more amino acid residues is according to formula IIa:

3. The polypeptide of claim 1 wherein the polypeptide incorporating one or more amino acid residues of formula IIa is selected from an immunoglobulin polypeptide, a V_H immunoglobulin polypeptide, a V_L immunoglobulin polypeptide, an Fv polypeptide, an Fab polypeptide, an (Fab′

)₂ polypeptide, a single chain Fv polypeptide, and an Fc polypeptide.

9. The polypeptide of claim 1, wherein the one or more amino acid residues of formula IIa is according to formula 30:

10. The polypeptide of claim 3, wherein the one or more amino acid residues of formula IIa is according to formula 30:

35. The polypeptide of claim 9 linked to a payload moiety via a triazole moiety and an optional linking moiety.

2. A polypeptide which is a polymer of amino acid residues where one or more amino acid residues is according to the following formula IIc:

4. The polypeptide of claim 2 wherein the polypeptide incorporating one or more amino acid residues of formula IIc is selected from an immunoglobulin polypeptide, a V_H immunoglobulin polypeptide, a V_L immunoglobulin polypeptide, an Fv polypeptide, an Fab polypeptide, an (Fab′

)₂ polypeptide, a single chain Fv polypeptide, and an Fc polypeptide.

12. The polypeptide of claim 2 where the payload is a polyethylene glycol, a cytotoxic compound, or a combination thereof.

13. The polypeptide of claim 2 prepared by contacting an alkyne-containing payload with a second polypeptide which is a polymer of amino acid residues where one or more amino acid residues is according to formula IIa:

14. The polypeptide of claim 13 where the alkyne-containing payload comprises an alkyne-containing PEG.

22. The polypeptide of claim 2 wherein W₄ is —

CH₂—

.

23. The polypeptide of claim 4 where the payload is a polyethylene glycol, a cytotoxic compound, or a combination thereof.

24. The polypeptide of claim 4 prepared by contacting an alkyne-containing payload with a second polypeptide which is a polymer of amino acid residues where one or more amino acid residues is according to formula IIa:

25. The polypeptide of claim 24 where the alkyne-containing payload comprises an alkyne-containing PEG.

27. The polypeptide of claim 25 which is linked to a payload by a heteroalkylene or substituted heteroalkylene linker.

28. The polypeptide of claim 27 where the linking moiety is a polyethylene glycol.

29. The polypeptide of claim 28 where the payload is a cytotoxic compound.

30. The polypeptide of claim 29 where the payload is a maytansinoid.

31. The polypeptide of claim 30 where the payload is maytansine.

32. The polypeptide of claim 30 where the payload is DM-1.

33. The polypeptide of claim 4 wherein W₄ is —

CH₂—

.

5. An tRNA aminoacylated with an amino acid residue according to formula IIb:

6. A method of producing a polypeptide, comprising contacting a first polypeptide which is a polymer of amino acids with the tRNA of claim 5 under conditions suitable for incorporating into the first polypeptide one or more amino acid residues of formula IIa:

7. The method of claim 6, wherein the tRNA is orthogonal and base pairs with a codon that is not normally associated with an amino acid.

8. The method of claim 6, wherein the contacting occurs in a reaction mixture which comprises a tRNA synthetase that aminoacylates the tRNA with a compound according to formula II:

11. The tRNA of claim 5, wherein formula IIb is according to the following formula:

34. The tRNA of claim 5 which is orthogonal and base pairs with a codon that is not normally associated with an amino acid.

15. A polypeptide which is a polymer of amino acid residues where one or more amino acid residues is according to the following formula:

16. The polypeptide of claim 15 which is linked to a payload by a heteroalkylene or substituted heteroalkylene linker.

17. The polypeptide of claim 16 where the linking moiety is a polyethylene glycol.

18. The polypeptide of claim 17 where the payload is a cytotoxic compound.

19. The polypeptide of claim 18 where the payload is a maytansinoid.

20. The polypeptide of claim 19 where the payload is maytansine.

21. The polypeptide of claim 19 where the payload is DM-1.

26. A polypeptide which is a polymer of amino acid residues where one or more amino acid residues is according to the following formula: