Heteromultimers with reduced or silenced effector function

Patent metadata

Field	Value
Publication number	US-2016102135-A1
Application number	US-201414893503-A
Country	US
Kind code	A1
Filing date	May 30, 2014
Priority date	May 31, 2013
Publication date	Apr 14, 2016
Grant date	—

Abstract

Provided herein are heteromultimer constructs with reduced or silenced effector function. In an embodiment is provided a heteromultimer construct comprising an IgG Fc construct having a first and a second Fc polypeptide, each Fc polypeptide comprising a modified lower hinge region wherein: the modified lower hinge region of said first Fc polypeptide comprises at least one amino acid modification, the modified lower hinge region of said second Fc polypeptide comprises at least one amino acid modification which is different from at least one amino acid modification of said first Fc polypeptide, and the IgG Fc construct displays reduced binding to all Fcγ receptors and to C1q protein as compared to a corresponding parent IgG Fc construct. Also provided are methods of producing such heteromultimer constructs, and methods of reducing ADCC for an antibody construct by reducing effector function.

First claim

1 . A heteromultimer comprising an IgG Fc construct having a first and a second Fc polypeptide, each Fc polypeptide comprising a modified lower hinge region wherein:
a. the modified lower hinge region of said first Fc polypeptide comprises at least one amino acid modification, b. the modified lower hinge region of said second Fc polypeptide comprises at least one amino acid modification which is different from at least one amino acid modification of said first Fc polypeptide, and c. the IgG Fc construct displays reduced binding to the FcγRIa, FcγRIIa, FcγRIIb and FcγRIIIa receptors as compared to a corresponding parent IgG Fc construct.

2 . The heteromultimer according to claim 1 , wherein
a. the at least one amino acid modification in the modified lower hinge region of the first Fc polypeptide increases the net positive charge in the modified lower hinge region, and the at least one amino acid modification in the second Fc polypeptide increases the total number of negative charges or is charge neutral relative to the parent hinge region; or b. the at least one amino acid modification in the modified lower hinge region of the first Fc polypeptide increases the net negative charge in the modified lower hinge region, and the at least one amino acid modification in the second Fc polypeptide increases the total number of positive charges.

3 . The heteromultimer according to claim 1 , wherein the modified lower hinge region of at least one of said first and second Fc polypeptides comprises two or more amino acid modifications.

4 . A heteromultimer comprising an IgG Fc construct having a first and a second Fc polypeptide, each Fc polypeptide comprising a modified hinge region wherein:
a. the modified hinge region of said first Fc polypeptide comprises at least one amino acid modification that increases the net charge in the modified hinge region of the first Fc polypeptide at about physiological pH conditions, b. the modified hinge region of said second Fc polypeptide comprises at least one amino acid modification which is different from at least one amino acid modification of said first Fc polypeptide, and c. the IgG Fc construct displays reduced binding to the FcγRIa, FcγRIIa, FcγRIIb and FcγRIIIa receptors as compared to a corresponding parent IgG Fc construct.

5 . The heteromultimer of claim 4 , wherein said increase in net charge is an increase in net positive charge due to an increase in the total number of positively charged amino acids on the first Fc polypeptide, or a decrease in the total number of negatively charged amino acids on the first Fc polypeptide.

6 . (canceled)

7 . The heteromultimer of claim 4 , wherein the at least one amino acid modification on the modified hinge region of said first Fc polypeptide increases the total number of positively charged amino acids on said first Fc polypeptide, and the at least one amino acid modification on the modified hinge region of said second Fc polypeptide increases the total number of negative charges on said second Fc polypeptide or is charge neutral.

8 . The heteromultimer of claim 4 , wherein said increase in net charge is an increase in the net negative charge due to an increase in the total number of negatively charged amino acids or a decrease in the total number of positively charged amino acids on the first Fc polypeptide.

9 . (canceled)

10 . The heteromultimer of claim 8 , wherein said increase in net charge is an increase in net negative charge due to an increase in the total number of negatively charged amino acids on the first Fc polypeptide, and the at least one amino acid modification on the second Fc polypeptide increases the total number of positive charges on the second Fc polypeptide.

11 . The heteromultimer according to claim 4 , wherein the at least one amino acid modification in the modified hinge region of the first Fc polypeptide combined with the at least one amino acid modification in the second Fc polypeptide increases the overall positive charge of the IgG Fc construct compared to the parent IgG Fc construct.

12 . The heteromultimer according to claim 1 , wherein the modified hinge region of at least one of said first and second Fc polypeptides comprises two or more amino acid modifications.

13 . The heteromultimer according to claim 1 , wherein the modified hinge region of each of said first and second Fc polypeptides comprises two or more amino acid modifications.

14 . (canceled)

15 . The heteromultimer according to claim 1 , wherein the IgG Fc construct has a K D of greater than 10 μM for FcγRIIaH, a K D of greater than 10 μM for FcγRIIaR, a K D of greater than 10 μM for FcγRIIb, a K D of greater than 6 μM for FcγRIIIaF, a K D of greater than 6 μM for FcγRIIIaV, and a K D of greater than 6.5 nM for FcγRIa.

16 . The heteromultimer according to claim 1 , wherein the IgG Fc construct mediates reduced effector function compared to the corresponding parent IgG Fc construct.

17 . (canceled)

18 . (canceled)

19 . The heteromultimer according to claim 16 , wherein the effector function is ADCC, ADCP, CDC or any combination thereof.

20 . The heteromultimer according to claim 1 , wherein the modified hinge region of at least one of said first and second Fc polypeptides comprises amino acid modifications at L234 and/or L235, wherein the numbering of amino acids is according to the EU index as in Kabat.

21 . (canceled)

22 . (canceled)

23 . (canceled)

24 . (canceled)

25 . The heteromultimer according to claim 20 , wherein said modification at L234 and/or L235 is selected from L234A, L234K, L234R, L234D, L234E, L235K, L235R, L235E, L235A, and L235D.

26 . The heteromultimer according to claim 20 , wherein the modified hinge region of the first and/or the second Fc polypeptide further comprises an amino acid modification at E233.

27 . The heteromultimer according to claim 26 , wherein either or both amino acid modifications at E233 are independently E233A, E233K, E233R, or E233D.

28 . The heteromultimer according to claim 1 , wherein the modified hinge region of at least one of said first and second Fc polypeptides comprises the amino acid modifications L234K/L235K, E233A/L234R/L235R, E233K/L234R/L235R, E233K/L234A/L235K, L234A/L235A, L234D/L235E, E233A/L234D/L235E, or E233A/L234K/L235A, wherein the numbering of amino acids is according to the EU index as in Kabat.

29 . (canceled)

30 . The heteromultimer according to claim 1 , wherein:
a. the modified hinge region of the first Fc polypeptide comprises the amino acid modifications L234K/L235K and the modified hinge region of the second Fc polypeptide comprises the amino acid modifications L234A/L235A; or b. the modified hinge region of the first Fc polypeptide comprises the amino acid modifications L234K/L235K and the modified hinge region of the second Fc polypeptide comprises the amino acid modifications L234D/L235E; or c. the modified hinge region of the first Fc polypeptide comprises the amino acid modifications E233A/L234R/L235R and the modified hinge region of the second Fc polypeptide comprises the amino acid modifications E233A/L234D/L235E; or d. the modified hinge region of the first Fc polypeptide comprises the amino acid modifications E233K/L234R/L235R and the modified hinge region of the second Fc polypeptide comprises the amino acid modifications

Assignees

• Zymeworks Inc

Inventors

• Escobar-Cabrera Eric

Classifications

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title

• A61P9/04

  Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P7/04

  Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents · CPC title

• A61P37/00

  Drugs for immunological or allergic disorders · CPC title