a2) United States Patent
`US 7,442,778 B2
`(0) Patent No.:
`Gegget al.
`Oct. 28, 2008
`(45) Date of Patent:
`
`US007442778B2
`
`(54)
`
`(75)
`
`MODIFIED FC MOLECULES
`
`Inventors: Colin Gegg, Newbury Park, CA (US);
`Fei Xiong, Thousand Oaks, CA (US);
`Karen C. Sitney, Weston, CT (US)
`
`(73) Assignee: Amgen Inc., Thousand Oaks, CA (US)
`
`(*) Notice:
`
`Subject to any disclaimer, the term ofthis
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 145 days.
`
`(21) Appl. No.: 11/234,731
`
`(22)
`
`Filed:
`
`Sep. 23, 2005
`
`(65)
`
`Prior Publication Data
`
`US 2006/0140934 Al
`
`Jun. 29, 2006
`
`Related U.S. Application Data
`
`(60) Provisional application No. 60/612,680,filed on Sep.
`24, 2004.
`
`(51)
`
`Int. Cl.
`(2006.01)
`C07K 16/00
`(2006.01)
`CO7H 21/02
`(52) US.CL wo. 530/391.7; 530/350; 530/391.1
`(58) Field of Classification Search «0.0.0.0... None
`See application file for complete search history.
`
`(56)
`
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`(Continued)
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`Primary Examiner—Sheela J Huff
`(74) Attorney, Agent, or Firm—Nisan A. Steinberg
`
`(57)
`
`ABSTRACT
`
`The present invention concerns molecules and a process in
`which one or more biologically active peptides are incorpo-
`rated into an Fe domain.In this invention, pharmacologically
`active compounds maybe prepared by a process comprising
`(a) selecting at least one peptide that modulates theactivity of
`a protein of interest; and (b) preparing a pharmacologic agent
`comprising an amino acid sequenceofthe selected peptide in
`a loop region of an Fc domain. This process may be employed
`to modify an Fe domainthatis already linked through an N-
`or C-terminus or sidechain to a peptide or to a polypeptide
`(e.g., etanercept). This process may also be employed to
`modify an Fe domain that is part of an antibody(e.g., adali-
`mumab, epratuzumab, infliximab, Herceptin®, andthe like).
`In this way, different molecules can be produced that have
`additional functionalities, such as a binding domain to a dif-
`ferent epitope or an additional binding domainto the precur-
`sor molecule’s existing epitope. The peptide can be selected,
`for example, by phage display, £. coli display, ribosome
`display, RNA-peptide screening, yeast-based screening,
`chemical-peptide screening, rational design, or protein struc-
`tural analysis.
`
`14 Claims, 22 Drawing Sheets
`
`APOTEX EX1046
`Page 1
`
`APOTEX EX1046
`
`Page 1
`
`

`

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`

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`U.S. Patent
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`Oct. 28, 2008
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`Sheet 1 of 22
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`US 7,442,778 B2
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`
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`U.S. Patent
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`Oct. 28, 2008
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`Sheet 2 of 22
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`U.S. Patent
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`Oct. 28, 2008
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`Sheet 3 of 22
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`U.S. Patent
`
`Oct. 28, 2008
`
`Sheet 4 of 22
`
`US 7,442,778 B2
`
`VeOla
`
`
`
`
`
`MHSAGAAAOLAUdLYSINILGN4dddaTHASddSTTadVdOddOLHLMGW
`
`
`
`
`
`
`
`
`
`AEMONIMGOHTALTIASAAUALSNAOZANdUNLMWNHATADGAAMNAMARaG
`
`
`
`
`
`
`
`ATOLISAONYLIHGUSddTLAACAMMAOOWWUSILWYAIAYaATWUNSANONM
`
`
`
`
`
`
`
`
`
`OOMUSUGALIMSATAAISOGSGIAMHLLNANNHGOONSAMAAVIGSdAJOM
`
`
`
`
`
`
`
`
`
`*M9dSISISMOLAHNHTVEHWASOSJAND
`
`TS
`
`TOT
`
`TST
`
`TO?
`
`Page 10
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`Page 10
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`

`

`U.S. Patent
`
`Oct. 28, 2008
`
`Sheet 5 of 22
`
`US 7,442,778 B2
`
`huFc-IgA2
`huFc-IgM
`huFc-IgGl