Modified amino acids

Patent metadata

Field	Value
Publication number	US-9682934-B2
Application number	US-201314015893-A
Country	US
Kind code	B2
Filing date	Aug 30, 2013
Priority date	Aug 31, 2012
Publication date	Jun 20, 2017
Grant date	Jun 20, 2017

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.

First claim

What is claimed is:

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

or a salt thereof, wherein W 4 is C 1 -C 10 alkylene.

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:

or a salt thereof, wherein W 4 is C 1 -C 10 alkylene; where the payload is a label; a poly(alkylene glycol); a polyethylene glycol; poly(dextran); poly(propylene glycol); X—CH 2 CH 2 O—(CH 2 CH 2 O) n —CH 2 CH 2 — or X—CH 2 CH 2 O—(CH 2 CH 2 O) n —CH 2 CH 2 —O—(CH 2 ) m —W— where n is 20 to 4000 and X is hydroxyl, alkenyl, amine, aminooxy, carboxylic acid, maleimide, dithiopyridine, or vinylpyridine; R z O—(CH 2 CH 2 O) n1 —O—(CH 2 ) m —, R z O—(CH 2 OCH 2 O) n1 —O—(CH 2 ) m1 —NH—C(O)—(CH 2 ) p1 —, or [R z O—(CH 2 CH 2 O) n1 —O—(CH 2 ) 2 —NH—C(O)] 2 CH(CH 2 ) m1 —X 10 —(CH 2 ) p1 — where R z is alkyl, m1 is 2-10, n1 is 100-1,000, p1 is 2-10, and X 10 is optionally an O, N, S or carbonyl group (C═O), or not present; or a cytotoxic compound; or a combination thereof.

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.

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.

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

or a salt thereof, wherein W 4 is C 1 -C 10 alkylene and R 1 is the remainder of a tRNA which is bonded to the amino acid residue through the 3′-OH or 2′-OH of a terminal ribose sugar of the tRNA.

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:

or a salt thereof, wherein W 4 is C 1 -C 10 alkylene.

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:

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

or a salt thereof.

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

or a salt thereof.

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

or a salt thereof.

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:

or a salt thereof, wherein W 4 is C 1 -C 10 alkylene.

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

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

or a salt thereof, wherein W 4 is C 1 -C 10 alkylene; where the linking moiety is a bifunctional, C 1-18 -alkylene, substituted C 1-18 -alkylene, hetero-C 1-18 -alkylene, substituted hetero-C 1-18 -alkylene, arylene, substituted arylene, heteroarylene or substituted heteroarylene linker; and where the payload is a label or a cytotoxic compound; or a combination thereof.

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.

22. The polypeptide of claim 2 wherein W 4 is —CH 2 —.

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:

or a salt thereof, wherein W 4 is C 1 -C 10 alkylene.

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

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

or a salt thereof, wherein W 4 is C 1 -C 10 alkylene; where the polypeptide 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; where the linking moiety is a bifunctional, C 1-18 -alkylene, substituted C 1-18 -alkylene, hetero-C 1-18 -alkylene, substituted hetero-C 1-18 -alkylene, arylene, substituted arylene, heteroarylene or substituted heteroarylene linker; and where the payload is a label or a cytotoxic compound, or a combination thereof.

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

28. The polype

Assignees

• Sutro Biopharma Inc

Inventors

• Stafford Ryan
• Thanos Christopher D
• Yang Wenjin

Classifications

• A61P37/02

  Immunomodulators · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P29/00

  Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID] · CPC title

• C07K14/535

  Granulocyte CSF; Granulocyte-macrophage CSF · CPC title

• C07C275/14

  being further substituted by nitrogen atoms not being part of nitro or nitroso groups · CPC title