Anti-PCSK9 antibodies and use thereof

• US 9,255,154 B2
• Filed: 03/12/2013
• Issued: 02/09/2016
• Est. Priority Date: 05/08/2012
• Status: Expired due to Fees

• Current Assignee: H. Lundbeck A/S (Lundbeck Foundation)
• Original Assignee: AlderBio Holdings, LLC (Lundbeck Foundation)
• Inventors: Feldhaus, Andrew Lawrence, Garcia-Martinez, Leon F., Dutzar, Benjamin H., Ojala, Ethan Wayne, Kovacevich, Brian Robert, Anderson, Katie Olson, Fan, Pei, Billgren, Jens, Stewart, Erica Ann, Akatsuka, Corinne C., McNeill, Patricia Dianne, Mitchell, Danielle Marie, Allison, Dan Scott, Latham, John A.
• Primary Examiner(s): WEN, SHARON X
• Application Number:
  US13/795,674
• Publication Number:
  US 20130302399A1
• Time in Patent Office: 1,064 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 Antihypertensives
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: G01N 2333/96433 Serine endopeptidases (3.4.21)
• CPC Class Codes: G01N 33/573 for enzymes or isoenzymes

Abstract

The present invention is directed to antibodies and fragments thereof having binding specificity for PCSK9. Another embodiment of this invention relates to the antibodies described herein, and binding fragments thereof, comprising the sequences of the V_H, V_L and CDR polypeptides described herein, and the polynucleotides encoding them. The invention also contemplates conjugates of anti-PCSK9 antibodies and binding fragments thereof conjugated to one or more functional or detectable moieties. The invention also contemplates methods of making said anti-PCSK9 antibodies and binding fragments thereof. Embodiments of the invention also pertain to the use of anti-PCSK9 antibodies, and binding fragments thereof, for the diagnosis, assessment and treatment of diseases and disorders associated with PCSK9.

Claims

1. An isolated antibody or antigen-binding fragment thereof that binds to proprotein convertase subtilisin/kexin type 9 (PCSK9), comprising a heavy chain and a light chain that each comprise complementarity determining regions (CDRs), wherein:

(a) the variable heavy chain comprises the CDR1 sequence of SEQ ID NO;

4, the CDR2 sequence of SEQ ID NO;

6, and the CDR3 sequence of SEQ ID NO;

8; and

the variable light chain comprises the CDR1 sequence of SEQ ID NO;

24, the CDR2 sequence of SEQ ID NO;

26, and the CDR3 sequence of SEQ ID NO;

28;

or

(b) the variable heavy chain comprises the CDR1 sequence of SEQ ID NO;

684, the CDR2 sequence of SEQ ID NO;

686, and the CDR3 sequence of SEQ ID NO;

688; and

the variable light chain comprises the CDR1 sequence of SEQ ID NO;

704, the CDR2 sequence of SEQ ID NO;

706, and the CDR3 sequence of SEQ ID NO;

708;

or

(c) the variable heavy chain comprises the CDR1 sequence of SEQ ID NO;

724, the CDR2 sequence of SEQ ID NO;

726, and the CDR3 sequence of SEQ ID NO;

728; and

the variable light chain comprises the CDR1 sequence of SEQ ID NO;

744, the CDR2 sequence of SEQ ID NO;

746, and the CDR3 sequence of SEQ ID NO;

748;

or

(d) the variable heavy chain comprises the CDR1 sequence of SEQ ID NO;

764, the CDR2 sequence of SEQ ID NO;

766, and the CDR3 sequence of SEQ ID NO;

768; and

the variable light chain comprises the CDR1 sequence of SEQ ID NO;

784, the CDR2 sequence of SEQ ID NO;

786, and the CDR3 sequence of SEQ ID NO;

788;

or

(e) the variable heavy chain comprises the CDR1 sequence of SEQ ID NO;

844, the CDR2 sequence of SEQ ID NO;

846, and the CDR3 sequence of SEQ ID NO;

848; and

the variable light chain comprises the CDR1 sequence of SEQ ID NO;

864, the CDR2 sequence of SEQ ID NO;

866, and the CDR3 sequence of SEQ ID NO;

868.

2. The antibody or antigen-binding fragment thereof of claim 1, wherein:

(i) the variable heavy chain comprises a sequence at least 90% identical to SEQ ID NO;

2, the variable light chain comprises a sequence at least 90% identical to SEQ ID NO;

22, and the variable heavy and light chains comprise the CDRs according to (a);

or

(ii) the variable heavy chain comprises a sequence at least 90% identical to SEQ ID NO;

682, the variable light chain comprises a sequence at least 90% identical to SEQ ID NO;

702, and the variable heavy and light chains comprise the CDRs according to (b);

or

(iii) the variable heavy chain comprises a sequence at least 90% identical to SEQ ID NO;

722, the variable light chain comprises a sequence at least 90% identical to SEQ ID NO;

742, and the variable heavy and light chains comprise the CDRs according to (c);

or

(iv) the variable heavy chain comprises a sequence at least 90% identical to SEQ ID NO;

762, the variable light chain comprises a sequence at least 90% identical to SEQ ID NO;

782, and the variable heavy and light chains comprise the CDRs according to (d);

or

(v) the variable heavy chain comprises a sequence at least 90% identical to SEQ ID NO;

842, the variable light chain comprises a sequence at least 90% identical to SEQ ID NO;

862, and the variable heavy and light chains comprise the CDRs according to (e).

3. The antibody or antigen-binding fragment thereof of claim 1, wherein:

(i) the variable heavy chain comprises a sequence at least 95% identical to SEQ ID NO;

2, the variable light chain comprises a sequence at least 95% identical to SEQ ID NO;

22, and the variable heavy and light chains comprise the CDRs according to (a);

or

(ii) the variable heavy chain comprises a sequence at least 95% identical to SEQ ID NO;

682, the variable light chain comprises a sequence at least 95% identical to SEQ ID NO;

702, and the variable heavy and light chains comprise the CDRs according to (b);

or

(iii) the variable heavy chain comprises a sequence at least 95% identical to SEQ ID NO;

722, the variable light chain comprises a sequence at least 95% identical to SEQ ID NO;

742, and the variable heavy and light chains comprise the CDRs according to (c);

or

(iv) the variable heavy chain comprises a sequence at least 95% identical to SEQ ID NO;

762, the variable light chain comprises a sequence at least 95% identical to SEQ ID NO;

782, and the variable heavy and light chains comprise the CDRs according to (d);

or

(v) the variable heavy chain comprises a sequence at least 95% identical to SEQ ID NO;

842, the variable light chain comprises a sequence at least 95% identical to SEQ ID NO;

862, and the variable heavy and light chains comprise the CDRs according to (e).

4. The antibody or antigen-binding fragment thereof of claim 1, wherein:

(i) the variable heavy chain comprises SEQ ID NO;

2, the variable light chain comprises SEQ ID NO;

22, and the variable heavy and light chains comprise the CDRs according to (a);

or

(ii) the variable heavy chain comprises SEQ ID NO;

682, the variable light chain comprises SEQ ID NO;

702, and the variable heavy and light chains comprise the CDRs according to (b);

or

(iii) the variable heavy chain comprises SEQ ID NO;

722, the variable light chain comprises SEQ ID NO;

742, and the variable heavy and light chains comprise the CDRs according to (c);

or

(iv) the variable heavy chain comprises SEQ ID NO;

762, the variable light chain comprises SEQ ID NO;

782, and the variable heavy and light chains comprise the CDRs according to (d);

or

(v) the variable heavy chain comprises SEQ ID NO;

842, the variable light chain comprises SEQ ID NO;

862, and the variable heavy and light chains comprise the CDRs according to (e).

5. The antibody or antigen-binding fragment thereof of claim 1, which is selected from the group consisting of chimeric, humanized, and human antibodies or antibody fragments, and scFvs, Fab fragments, Fab′

fragments, monovalent antibody fragments, and F(ab′

)2 fragments.

6. The antibody or antigen-binding fragment thereof of claim 1, which lacks N-glycosylation and/or O-glycosylation.

7. The antibody or antigen-binding fragment thereof of claim 1, which comprises a human constant domain.

8. The antibody or antigen-binding fragment thereof of claim 7, wherein said human constant domain comprises an IgG1, IgG2, IgG3, or IgG4 constant domain.

9. The antibody or antigen-binding fragment thereof of claim 1, which comprises an Fc region that has been modified to alter at least one of effector function, half-life, proteolysis, or glycosylation.

10. The antibody or antigen-binding fragment thereof of claim 9, wherein the Fc region contains one or more mutations that alters or eliminates N- and/or O-glycosylation.

11. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody or antigen-binding fragment thereof is humanized.

12. The antibody or antigen-binding fragment thereof of claim 1, which when administered to a human subject inhibits or neutralizes at least one biological effect elicited by PCSK9.

13. The antibody or antigen-binding fragment thereof of claim 1, which, when administered to a human subject, reduces serum cholesterol and/or inhibits the binding of PCSK9 to LDLR.

14. The antibody or antigen-binding fragment thereof of claim 1, which binds to human PCSK9 with a K_D that is less than about 100 nM.

15. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody or antigen-binding fragment thereof is attached to one or more detectable moieties.

16. The antibody or antigen-binding fragment thereof of claim 15, wherein the detectable moiety comprises a fluorescent dye, enzyme, substrate, bioluminescent material, radioactive material, chemiluminescent moiety, or mixtures thereof.

17. A pharmaceutical composition comprising at least one antibody or antigen-binding fragment thereof according to claim 1 and a pharmaceutically acceptable diluent, carrier, solubilizer, emulsifier, preservative, or mixture thereof.

18. The pharmaceutical composition of claim 17, further comprising another active agent.

19. The pharmaceutical composition of claim 17, wherein the other active agent is selected from the group consisting of the following:

statins, ACE inhibitors, angiotensin II receptor blockers (ARBs), antiarrhythmics, antiplatelet drugs, aspirin, beta blockers, amiodarone, digoxin, aspirin, anti-clotting agents, digoxin, diuretics, heart failure drugs, vasodilators, blood thinners, anti-cholesterol drugs, cholestyramine, gemfibrozil, omega-3 polyunsaturated fatty acids, pantethine, anti-hypertensives, antidiabetogenic drugs, meglitinides, sulfonylurea, and thiazolidinediones.

20. The pharmaceutical composition of claim 19, which is lyophilized, stabilized and/or or formulated for administration by injection.