Multispecific antigen-binding proteins

Patent metadata

Field	Value
Publication number	US-11116840-B2
Application number	US-201715789670-A
Country	US
Kind code	B2
Filing date	Oct 20, 2017
Priority date	Apr 24, 2015
Publication date	Sep 14, 2021
Grant date	Sep 14, 2021

Abstract

Provided are, inter alia, multispecific antigen binding proteins, or antigen-binding fragments thereof, comprising one or more mutations in the VH/VL domains and/or CH1/CL domains, pharmaceutical compositions comprising same, isolated nucleic acids, vectors, and host cells encoding/expressing same, method of making the multispecific antigen binding proteins, computer readable media for evaluating multispecific antigen binding proteins, and libraries.

First claim

What is claimed is:

1. A multispecific antigen binding protein comprising: a) a first heavy chain/light chain pair binding to a first antigen which comprises a first heavy chain polypeptide (H1) and a first light chain polypeptide (L1), and b) a second heavy chain/light chain pair binding to a second antigen which comprises a second heavy chain polypeptide (H2) and a second light chain polypeptide (L2), wherein each H1 and H2 comprises a heavy chain variable domain (VH) and a heavy chain constant domain (CH1), and each L1 and L2 comprises a light chain variable domain (VL) and a light chain constant domain (CL);
wherein:
(i) an amino acid at S183 (EU numbering) in the CH1 domain of H1 is replaced with a positively charged residue, an amino acid at Q39 (Kabat numbering) in the VH domain of H1 is replaced with a negatively charged residue, an amino acid at V133 (EU numbering) in the CL domain of L1 is replaced with a negatively charged residue, and an amino acid at Q38 (Kabat numbering) in the VL domain of L1 is replaced with a positively charged residue; and
an amino acid at Q39 (Kabat numbering) in the VH domain of H2 is replaced with a positively charged residue and an amino acid at Q38 (Kabat numbering) in the VL domain of L2 is replaced with a negatively charged residue;

or
(ii) an amino acid at S183 (EU numbering) in the CH1 domain of H1 is replaced with a negatively charged residue, an amino acid at Q39 (Kabat numbering) in the VH domain of H1 is replaced with a positively charged residue, an amino acid at V133 (EU numbering) in the CL domain of L1 is replaced with a positively charged residue, and an amino acid at Q38 (Kabat numbering) in the VL domain of L1 is replaced with a negatively charged residue; and
an amino acid at Q39 (Kabat numbering) in the VH domain of H2 is replaced with a negatively charged residue and an amino acid at Q38 (Kabat numbering) in the VL domain of L2 is replaced with a positively charged residue;
wherein the positively charged residue is selected from the group consisting of R and K and wherein the negatively charged residue is selected from the group consisting of D and E.

2. The multispecific antigen binding protein of claim 1 , wherein the CH1 domain consists of an amino acid substitution at S183 (EU numbering), and wherein the CL domain of L1 consists of an amino acid substitution at V133 (EU numbering).

3. The multispecific binding antigen of claim 1 ,
wherein the amino acid at S183 (EU numbering) in the CH1 domain of H1 is replaced with a positively charged residue, the amino acid at Q39 (Kabat numbering) in the VH domain of H1 is replaced with a negatively charged residue, the amino acid at V133 (EU numbering) in the CL domain of L1 is replaced with a negatively charged residue, and the amino acid at Q38 (Kabat numbering) in the VL domain of L1 is replaced with a positively charged residue; the amino acid at Q39 (Kabat numbering) in the VH domain of H2 is replaced with a positively charged residue, and the amino acid at Q38 (Kabat numbering) in the VL domain of L2 is replaced with a negatively charged residue; and
wherein the amino acid at S183 (EU numbering) in the CH1 domain of H2 is replaced with a negatively charged amino acid residue, and the amino acid at V133 (EU numbering) in the CL domain of L2 is replaced with a positively charged amino acid residue.

4. The multispecific antigen binding protein of claim 1 ,
wherein the amino acid at S183 (EU numbering) in the CH1 domain of H1 is replaced with a negatively charged residue, the amino acid at Q39 (Kabat numbering) in the VH domain of H1 is replaced with a positively charged residue, the amino acid at V133 (EU numbering) in the CL domain of L1 is replaced with a positively charged residue, and the amino acid at Q38 (Kabat numbering) in the VL domain of L1 is replaced with a negatively charged residue; the amino acid at Q39 (Kabat numbering) in the VH domain of H2 is replaced with a negatively charged residue and the amino acid at Q38 (Kabat numbering) in the VL domain of L2 is replaced with a positively charged residue; and
wherein the amino acid at S183 (EU numbering) in the CH1 domain of H2 is replaced with a positively charged residue, and the amino acid at V133 (EU numbering) in the CL domain of L2 is replaced with a negatively charged residue.

5. The multispecific antigen binding protein of claim 3 , wherein the CH1 domain of H2 consists of an amino acid substitution at S183 (EU numbering), and wherein the CL domain of L2 consists of an amino acid substitution at V133 (EU numbering).

6. The multispecific binding antigen of claim 1 , wherein
(a) the CH1 domain of H2 comprises the A141I, F170S, S181M, S183V, and V185A mutations, and the CL domain of L2 comprises the F116A, V133I, L135V, S162M, S174A, S176F, and T178V mutations;
(b) the CH1 domain of H2 comprises the A141I, F170S, S181M, S183A, and V185A mutations and the CL domain of L2 comprises the F116A, S131D, L135V, S162A, S174A, S176F, and T178I mutations;
(c) the CH1 domain of H2 comprises the A141I, F170S, S181M, S183A, and V185A mutations and the CL domain of L2 comprises the F116A, L135V, S174A, S176F, and T178V mutations;
(d) the CH1 domain of H2 comprises the A141I, F170A, S181M, S183V, and V185A mutations and the CL domain of L2 comprises the F116A, L135V, S162M, S174A, S176F, and T178V mutations;
(e) the CH1 domain of H2 comprises L128F, A141M, F170M, S181I and S183A mutations and the CL domain of L2 comprises F118V, S131T, V133A, L135Y, S162A, T164S, S176M, and T178L mutations;
(f) the CH1 domain of H2 comprises L128F, A141M, F170Y, S181I, S183A, and V185A mutations and the CL domain of L2 comprises F118V, S131T, V133A, L135F, S162A, S176A, and T178L mutations;
(g) the CH1 domain of H2 comprises L128F, A141T, F170M, S181T, S183A, and V185L mutations and the CL domain of L2 comprises F118V, S131T, V133A, L135F, S162A, T164S, S176T, and T178L mutations; or
(h) the CH1 domain of H2 comprises L128F, A141M, F170M, S181T, and S183A, mutations and the CL domain of L2 comprises F118V, S131T, V133A, L135F, S162M, T164S, S176M, and T178L mutations.

7. The multispecific antigen binding protein of claim 1 ,
wherein the amino acid at S183 (EU numbering) in the CH1 domain of H1 is replaced with a negatively charged residue, the amino acid at Q39 (Kabat numbering) in the VH domain of H1 is replaced with a positively charged residue, the amino acid at V133 (EU numbering) in the CL domain of L1 is replaced with a positively charged residue, the amino acid at Q38 (Kabat numbering) in the VL domain of L1 is replaced with a negatively charged residue, the amino acid at Q39 (Kabat numbering) in the VH domain of H2 is replaced with a negatively charged residue, and the amino acid at Q38 (Kabat numbering) in the VL domain of L2 is replaced with a positively charged residue;
wherein the VH domain of H1 comprises a Q39K mutation, the CH1 domain of H1 comprises an S183E mutation, the VL domain of L1 comprises a Q38E mutation, the CL domain of L1 comprises a V133K mutation, VH domain of H2 comprises a Q39E mutation, and the VL domain of L2 comprises a Q38K mutation.

8. The multispecific antigen binding protein of claim 7 , wherein the CH1 domain of H2 comprises a S183K mutation and the CL domain of L2 comprises the V133E mutation.

9. The multispecific antigen binding protein of claim 7 , wherein the CH1 domain of H2 comprises the A141I, F170S, S181M, S183A, and V185A mutations and the CL domain of L2 comprises the F116A, L135V, S174A, S176F, and T178V mutations.

10. The multispecific antigen binding protein of claim 1 ,
wherein the amino acid at S183 (EU numbering) in the CH1 domain of H1 is replaced with a positively charged residue, the amino acid at Q39 (Kaba

Assignees

• Genentech Inc

Inventors

• Carter Paul
• Spiess Christoph
• Yin Yiyuan
• Zhou Jianhui
• Sandoval Wendy
• Corn Jacob
• Dillon Michael

Classifications

• A61K2039/505

  comprising antibodies · CPC title

• A61P37/02

  Immunomodulators · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• C07K2317/31

  multispecific · CPC title

• C07K2317/56

  variable (Fv) region, i.e. VH and/or VL · CPC title