Anti-PCSK9 antibodies with pH-dependent binding characteristics

Patent metadata

Field	Value
Publication number	US-9540449-B2
Application number	US-201313964159-A
Country	US
Kind code	B2
Filing date	Aug 12, 2013
Priority date	Aug 13, 2012
Publication date	Jan 10, 2017
Grant date	Jan 10, 2017

Abstract

The present invention provides antibodies and antigen-binding fragments thereof that specifically bind proprotein convertase subtilisin/kexin-9 (PCSK9) with greater affinity at neutral pH than at acidic pH. The antibodies of the invention may possess one or more amino acid changes as compared to antibodies that do not exhibit pH-dependent binding properties. For example, the present invention includes anti-PCSK9 antibodies which possess one or more histidine substitutions in one or more complementarity determining regions. The antibodies of the invention, with pH-dependent binding properties, remain in circulation and exhibit cholesterol lowering activity for prolonged periods of time in animal subjects as compared to anti-PCSK9 antibodies that do not exhibit pH-dependent binding properties. The antibodies of the invention are therefore useful for treating diseases and disorders related to elevated HDL cholesterol, wherein the antibodies of the invention can be administered to a patient at a lower dose and/or with less frequent dosing as compared to antibodies that do not exhibit pH-dependent binding properties.

First claim

What is claimed is:

1. An isolated antibody or antigen-binding fragment thereof that binds human proprotein convertase subtilisin/kexin type 9 (PCSK9), wherein the acidic/neutral K D ratio for the antibody or antigen-binding fragment binding to PCSK9 at 25° C. is greater than about 12.5 as determined by surface plasmon resonance; wherein the antibody or antigen-binding fragment thereof comprises 3 heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) and 3 light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3),
wherein the HCDR1 comprises the amino acid sequence of SEQ ID NO: 220;
wherein the HCDR2 comprises the amino acid sequence of SEQ ID NO: 222;
wherein the HCDR3 comprises the amino acid sequence selected from the group consisting of SEQ ID NOs: 224 and 788;
wherein the LCDR1 comprises the amino acid sequence of SEQ ID NO: 802;
wherein the LCDR2 comprises the amino acid sequence of SEQ ID NO: 230; and
wherein the LCDR3 comprises the amino acid sequence of SEQ ID NO: 232.

2. The isolated antibody or antigen-binding fragment of claim 1 , wherein the acidic/neutral k d ratio for the antibody or antigen-binding fragment binding to PCSK9 at 25° C. is greater than about 7.5 as determined by surface plasmon resonance.

3. The isolated antibody or antigen-binding fragment of claim 1 , wherein the acidic/neutral t1/2 ratio for the antibody or antigen-binding fragment binding to PCSK9 at 25° C. is less than about 0.14 as determined by surface plasmon resonance.

4. The isolated antibody or antigen-binding fragment of claim 3 , wherein the antibody or antigen-binding fragment thereof binds human proprotein convertase subtilisinfkexin type 9 (PCSK9) at 25° C. and acidic pH with a dissociative half-life (t1/2) less than about 4.5 minutes, wherein the antibody or antigen-binding fragment thereof binds PCSK9 at 25° C. and neutral pH with a t1/2 of greater than about 35 minutes.

5. The isolated antibody or antigen-binding fragment of claim 4 , wherein the antibody or antigen-binding fragment thereof binds PCSK9 at 25° C. and acidic pH with a dissociative half-life (t1/2) less than about 2 minutes, wherein the antibody or antigen-binding fragment thereof binds PCSK9 at 25° C. and neutral pH with a t1/2 of greater than about 35 minutes.

6. The isolated antibody or antigen-binding fragment of claim 5 , wherein the antibody or antigen-binding fragment thereof binds PCSK9 at 25° C. and acidic pH with a dissociative half-life (t1/2) less than about 1.5 minutes, wherein the antibody or antigen-binding fragment thereof binds PCSK9 at 25° C. and neutral pH with a t1/2 of greater than about 35 minutes.

7. The isolated antibody or antigen-binding fragment of claim 1 , wherein the antibody or antigen-binding fragment thereof blocks the interaction between human proprotein convertase subtilisin/kexin type 9 (PCSK9) and the low density lipoprotein receptor (LDLR) at neutral pH with an IC 50 that is at least 36 times less than the PCSK9/LDLR blocking IC 50 value of the antibody or antigen-binding fragment thereof at acidic pH.

8. The isolated antibody or antigen-binding fragment of claim 1 , wherein the antibody or antigen-binding fragment thereof, when administered to a subject in a single dose of about 10 mg/kg, reduces serum LDL-C by at least 33% from baseline, and wherein the reduction in serum LDL-C is sustained for at least 26 days after administration.

9. The isolated antibody or antigen-binding fragment of claim 8 , wherein the antibody or antigen-binding fragment thereof, when administered to a subject in a single dose of about 10 mg/kg, reduces serum LDL-C by at least 33% from baseline, and wherein the reduction in serum LDL-C is sustained for at least 33 days after administration.

10. The isolated antibody or antigen-binding fragment of claim 1 , wherein the antibody or antigen-binding fragment thereof, when administered to a subject in a single dose of about 10 mg/kg, reduces serum LDL-C by at least 15% from baseline, and wherein the reduction in serum LDL-C is sustained for at least 42 days after administration.

11. The isolated antibody or antigen-binding fragment of claim 10 , wherein the antibody or antigen-binding fragment thereof, when administered to a subject in a single dose of about 10 mg/kg, reduces serum LDL-C by at least 15% from baseline, and wherein the reduction in serum LDL-C is sustained for at least 55 days after administration.

12. The isolated antibody or antigen-binding fragment of claim 1 , wherein the HCDR3 comprises the amino acid sequence of SEQ ID NO:224.

13. The isolated antibody or antigen-binding fragment of claim 1 , wherein the HCDR3 comprises the amino acid sequence of SEQ ID NO:788.

Assignees

• Regeneron Pharma

Inventors

• Yancopoulos George D
• Papadopoulos Nicholas J
• Murphy Andrew J
• Stahl Neil

Classifications

• C07K2317/94

  Stability, e.g. half-life, pH, temperature or enzyme-resistance · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C07K2317/76

  Antagonist effect on antigen, e.g. neutralization or inhibition of binding · CPC title

• C07K2317/21

  from primates, e.g. man · CPC title