Modified Fc molecules

• US 9,114,175 B2
• Filed: 06/28/2011
• Issued: 08/25/2015
• Est. Priority Date: 08/12/2005
• Status: Active Grant

• Current Assignee: Amgen Inc.
• Original Assignee: Amgen Inc.
• Inventors: Walker, Kenneth W., Miranda, Leslie P., Gegg, Colin V., Xiong, Fei
• Primary Examiner(s): Kim, Yunsoo
• Application Number:
  US13/171,233
• Publication Number:
  US 20120009205A1
• Time in Patent Office: 1,519 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 38/02 Peptides of undefined number of amino acids; Derivatives thereof
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/68 the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
• CPC Class Codes: A61K 47/6817 Toxins
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: A61P 9/12 Antihypertensives
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 17/00 Carrier-bound or immobilised peptides carrier-bound or immobilised enzymes C12N11/00; Preparation thereof
• CPC Class Codes: C07K 2317/40 characterized by post-translational modification
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype

Abstract

Disclosed is a process for preparing a pharmacologically active compound, in which at least one internal conjugation site of an Fc domain sequence is selected that is amenable to conjugation of an additional functional moiety by a defined conjugation chemistry through the side chain of an amino acid residue at the conjugation site. An appropriate amino acid residue for conjugation may be present in a native Fc domain at the conjugation site or may be added by insertion (i.e., between amino acids in the native Fc domain) or by replacement (i.e., removing amino acids and substituting different amino acids). In the latter case, the number of amino acids added need not correspond to the number of amino acids removed from the previously existing Fc domain. This technology may be used to produce useful compositions of matter and pharmaceutical compositions containing them. A DNA encoding the inventive composition of matter, an expression vector containing the DNA, and a host cell containing the expression vector are also disclosed.

Claims

1. A composition of matter of the formula

2. The composition of matter of claim 1, wherein the Fc domain comprises an IgG Fc domain.

3. The composition of matter of claim 1, wherein the Fc domain comprises an IgG1 Fc domain.

4. The composition of matter of claim 3, wherein the IgG1 Fc domain comprises SEQ ID NO:

600 or SEQ ID NO;

603.

5. The composition of matter of claim 1, wherein a =0 and b=1, or a=1 and b=0, or a=1 and b=1, or a=0 and b=0.

6. The composition of matter of claim 1, wherein X³ comprises polyethylene glycol (PEG).

7. The composition of matter of claim 1, wherein X³ has the structure -(L¹)_c-P¹ or -(L¹)_c-P¹-(L²)_d-P².

8. The composition of matter of claim 1, wherein g=1 or 2.

9. The composition of matter of claim 1, wherein the pharmacologically active peptide is a toxin peptide and/or comprises a cyclic peptide.

10. The composition of matter of claim 1, wherein P⁰, P¹, P², P³, or P⁴ is a non-peptide organic moiety.

11. The composition of matter of claim 1, wherein P⁰, P¹, P², P³, or P⁴ is a radioisotope, an enzyme, a biotinyl moiety, a fluorophore, or a chromophore.

12. The composition of matter of claim 1, wherein P⁰, P¹, P², P³, or P⁴ is selected from the group consisting of an immobilized substrate, a half-life extending moiety, a polymer, and polyethylene glycol.

13. The composition of matter of claim 12, wherein the half-life extending moiety is a polyethylene glycol, a copolymer of ethylene glycol, a polypropylene glycol, a copolymer of propylene glycol, a carboxymethylcellulose, a polyvinyl pyrrolidone, a poly-1,3-dioxolane, a poly-1,3,6-trioxane, an ethylene maleic anhydride copolymer, a polyaminoacid, a dextran n-vinyl pyrrolidone, a poly n-vinyl pyrrolidone, a propylene glycol homopolymer, a propylene oxide polymer, an ethylene oxide polymer, a polyoxyethylated polyol, a polyvinyl alcohol, a linear or branched glycosylated chain, a polyacetal, a long chain fatty acid, a long chain hydrophobic aliphatic group, an immunoglobulin F_c domain, an albumin, a transthyretin, or a thyroxine-binding globulin;

or a combination of any of these members.

14. The composition of matter of claim 1, wherein any of X¹, X², or X³ comprises a B cell activating factor (BAFF) binding peptide or polypeptide.

15. The composition of matter of claim 14, wherein the BAFF binding peptide or polypeptide comprises the amino acid sequence of SEQ ID NO:

514.

16. A pharmaceutical composition, comprising the composition of matter of claim 1, and a pharmaceutically acceptable carrier.

17. The composition of matter of claim 1, wherein:

(a) the composition of matter is an antibody modified so that it comprises at least one X³ additional functional moiety covalently bound to the Fc domain of the antibody through one or more specifically selected internal conjugation site(s) in the Fc domain through the side chain of an amino acid residue selected from the group consisting of D46, S48, H49, E50, E53, K55, D61, G62, Q76, K107, K121, G122, Q123, R136, D137, T140, K141, N142, N170, K173, S181, G183, D194, K195, R197, and Q199, relative to reference sequence SEQ ID NO;

599, or, if there is more than one X³, any combination of these members;

(b) X³ is selected from -(L¹)_c-P⁰, -(L¹)_c-P¹, -(L¹)_c-P¹-(L²)_d-P², -(L¹)_c-P¹-(L²)_d-P²-(L³)_e-P³, and -(L¹)_c-P¹-( L²)_d-P²-(L³)_e-P³-(L⁴)_f-P⁴;

P⁰, P¹, P², P³, and P⁴ are each independently selected from the group consisting of;

i) a pharmaceutically acceptable polymer or dextran;

ii) a pharmacologically active polypeptide, peptide, peptidomimetic, or non-peptide organic moiety;

iii) a radioisotope, an enzyme, a biotinyl moiety, a fluorophore, or a chromophore; and

iv) an immobilized substrate, provided that in a chain comprising more than one additional functional moieties, the immobilized substrate is the moiety most distal from the Fc domain, and there can be no more than one immobilized substrate in the chain;

L¹, L², L³, and L⁴ are each independently linkers;

c, d, e, and f are each independently 0 or 1.

18. A pharmaceutical composition, comprising the composition of matter of claim 1, and a pharmaceutically acceptable carrier.

19. A pharmaceutical composition, comprising the composition of matter of claim 17, and a pharmaceutically acceptable carrier.

20. The composition of matter of claim 17, comprising a B cell activating factor (BAFF) binding peptide or polypeptide.

21. The composition of matter of claim 20, wherein the BAFF binding peptide or polypeptide comprises the amino acid sequence of SEQ ID NO:

514.