LRP-8 binding dual variable domain immunoglobulin proteins

• US 10,093,733 B2
• Filed: 12/11/2015
• Issued: 10/09/2018
• Est. Priority Date: 12/11/2014
• Status: Active Grant

• Current Assignee: Abbvie Incorporated
• Original Assignee: Abbvie Incorporated
• Inventors: Benatuil, Lorenzo, Deng, Kangwen, Hanzatian, Denise Karaoglu, Argiriadi, Maria A.
• Primary Examiner(s): Howard, Zachary C
• Application Number:
  US14/967,260
• Publication Number:
  US 20160200813A1
• Time in Patent Office: 1,033 Days
• Field of Search: None
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• 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: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• 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 2333/705 Assays involving receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds

Abstract

The disclosure provides binding proteins that specifically bind to LRP-8 and optionally cross the blood brain barrier (BBB), localize to the brain and/or localize to the spinal cord.

Claims

1. A dual variable domain immunoglobulin (DVD-Ig) binding protein capable of binding low density lipoprotein receptor-related protein 8 (LRP-8), comprising a heavy chain variable domain comprising a set of three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2, and CDR-H3), and a light chain variable domain comprising a set of three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3), wherein the heavy chain CDRs are:

(a) CDR-H1;

RFTFSNX₁GMS (SEQ ID NO;

     9), wherein X₁ is F or Y, or GFTVSDYYMA (SEQ ID NO;

     53);

(b) CDR-H2;

TISSGGRYTYYPDX₂VKG (SEQ ID NO;

     10), wherein X₂ is S or H, or SISYEGSSTYYGDSVKG (SEQ ID NO;

     54); and

(c) CDR-H3;

DYLYAMDY (SEQ ID NO;

     46) or PLRYYGYNYRFAY (SEQ ID NO;

     55), and the light chain CDRs are;

(d) CDR-L1;

RSSQSLVYSX₃X₄NTYLH (SEQ ID NO;

     47), wherein X₃ is N, T, R, W or P, and wherein X₄ is G, E, L or K, or KASQNIHKNLD (SEQ ID NO;

     56);

(e) CDR-L2;

KVSNRFS (SEQ ID NO;

     48) or YTDNLQT (SEQ ID NO;

     57); and

(f) CDR-L3;

SQSTHVPLT (SEQ ID NO;

     49) or YQYNSGPT (SEQ ID NO;

     58).

2. The DVD-Ig binding protein of claim 1, wherein the set of three heavy chain CDRs is selected from the group consisting of:

(a) SEQ ID NO;

88, SEQ ID NO;

89, SEQ ID NO;

90;

(b) SEQ ID NO;

91, SEQ ID NO;

89, SEQ ID NO;

90;

(c) SEQ ID NO;

92, SEQ ID NO;

93, SEQ ID NO;

46;

(d) SEQ ID NO;

92, SEQ ID NO;

94, SEQ ID NO;

46; and

(e) SEQ ID NO;

107, SEQ ID NO;

108, SEQ ID NO;

109;

wherein the set of three light chain CDRs is selected from the group consisting of;

(i) SEQ ID NO;

157, SEQ ID NO;

48, SEQ ID NO;

49;

(ii) SEQ ID NO;

158, SEQ ID NO;

48, SEQ ID NO;

49;

(iii) SEQ ID NO;

159, SEQ ID NO;

48, SEQ ID NO;

49;

(iv) SEQ ID NO;

160, SEQ ID NO;

48, SEQ ID NO;

49;

(v) SEQ ID NO;

161, SEQ ID NO;

48, SEQ ID NO;

49;

(vi) SEQ ID NO;

162, SEQ ID NO;

48, SEQ ID NO;

49;

(vii) SEQ ID NO;

163, SEQ ID NO;

48, SEQ ID NO;

49;

(viii) SEQ ID NO;

164, SEQ ID NO;

48, SEQ ID NO;

49; and

(xi) SEQ ID NO;

56, SEQ ID NO;

57, SEQ ID NO;

58.

3. The DVD-Ig binding protein of claim 1, wherein:

(a) the heavy chain variable domain comprises an amino acid sequence of EVQLVESGGDLVKPGGSLKLSCAASRFTFSNFGMSWVRQTPDKRLEWV ATISSGGRYTYYPDX₁VKGRFTISRDNAKNTLYLQMSSLRSEDTAMYY CARDYLYAMDYWGQGTSVTVSS (SEQ ID NO;

     50), or EVQLVESGGDLVKPGGSLKLSCAASRFTFSNYGMSWVRQTPDKRLEWV ATISSGGRYTYYPDX₁VKGRFTISRDNAKNTLYLQMSSLRSEDTAMYY CARDYLYAMDYWGQGTSVTVSS (SEQ ID NO;

     51), wherein X₁ is S or H;

or EVQLVESGGGINQPGRSLKLSCAASGFTVSDYYMAWVRQAPKKGLEWV ASISYEGSSTYYGDSVKGRFTISRDNAKSILYLQMNSLRSEDTATYYC ARPLRYYGYNYRFAYWGQGTLVTVSS (SEQ ID NO;

     59), and/or

(b) the light chain variable domain comprises an amino acid sequence of DVVMTQTPLSLPVSLGDQASISCRSSQSLVYSX₂X₃NTYLHWYLQKPG QSPKVLMYKVSNRFSGVSDRFSGSGSGTDFTLKISRVEAEDLGVYFCS QSTHVPLTFGAGTKLELK (SEQ ID NO;

     52) wherein X₂ is N, T, R, W or P and X₃ is G, E, L or K;

or DIQMSQSPPVLSASVGDRVILSCKASQNIHKNLDWYQQKHGEAPKLLI IYYTDNLQTGIPSRFSGSGSGTDYTLTISSLQPEDVATYYCYQYNSGP TFGAGTKLELQ (SEQ ID NO;

     60).

4. The DVD-Ig binding protein of claim 1, wherein the binding protein comprises a heavy chain CDR set of:

(a) CDR-H1;

RFTFSNYGMS (SEQ ID NO;

     92);

(b) CDR-H2;

TISSGGRYTYYPDSVKG (SEQ ID NO;

     93); and

(c) CDR-H3;

DYLYAMDY (SEQ ID NO;

     46),

and a light chain CDR set of;

(d) CDR-L1;

RSSQSLVYSNGNTYLH (SEQ ID NO;

     157);

(e) CDR-L2;

KVSNRFS (SEQ ID NO;

     48); and

(f) CDR-L3;

SQSTHVPLT (SEQ ID NO;

     49).

5. The DVD-Ig binding protein of claim 1, wherein the binding protein comprises

(a) a heavy chain variable domain of:

6. The DVD-Ig binding protein of claim 1, wherein the binding protein is also capable of binding a second antigen target in the brain or spinal cord other than LRP-8.

7. The DVD-Ig binding protein of claim 1, wherein the binding protein binds human LRP-8 and is cross-reactive with at least one of cynomolgus monkey, rat, and mouse LRP-8.

8. The DVD-Ig binding protein of claim 1, wherein the binding protein

(a) binds to human LRP-8 with an EC50 of at most about 2.62 nM, or at most about 2.29 nM, or at most about 0.88 nM, or at most about 0.24 nM;

(b) undergoes endocytosis by HEK293 cells expressing LRP-8 at a rate between 1.5 and 2.5 times the endocytosis of control IgG into HEK293 cells expressing LRP-8;

(c) undergoes transcytosis across a Caco-2 cell monolayer expressing LRP-8 at a rate between 1.5 and 2.0 times the transcytosis of control IgG across a Caco-2 cell monolayer expressing LRP-8; and

/or

(d) is able to cross the blood-brain barrier after binding to LRP-8.

9. The DVD-Ig binding protein of claim 1, further comprising a wild-type human lambda or kappa light chain constant region, and a wild-type human IgG1 heavy chain constant region or a variant of a human IgG1 heavy chain constant region.

10. The DVD-Ig binding protein of claim 9, wherein the variant comprises a leucine (L) to alanine (A) mutation at amino acid positions 234 and 235 of human IgG1.

11. The DVD-Ig binding protein of claim 1, further comprising a heavy chain constant region selected from the group consisting of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, and IgD, or a variant thereof.

12. The DVD-Ig binding protein of claim 1, wherein the heavy chain variable domain comprises an amino acid sequence at least about 80%, 90%, 95%, or 98% homologous to any of those selected from SEQ ID NOs:

59, 61, 62, and 70-72, and/or wherein the light chain variable domain comprises an amino acid sequence at least about 80%, 90%, 95%, or 98% homologous to any of those selected from SEQ ID NOs;

60, 63, and 133-142.

13. The DVD-Ig binding protein of claim 1, wherein the heavy chain variable domain comprises an amino acid sequence selected from SEQ ID NOs:

59, 61, 62, and 70-72 and/or wherein the light chain variable domain comprises an amino acid sequence selected from SEQ ID NOs;

60, 63, and 133-142.

14. A conjugate comprising the DVD-Ig binding protein of claim 1 and a second agent, wherein the second agent acts on the nervous system and is capable of being transported across the blood-brain barrier when the conjugate binds to LRP-8.

15. A pharmaceutical composition comprising the DVD-Ig binding protein of claim 1, and a pharmaceutically acceptable carrier.

16. The pharmaceutical composition of claim 15, further comprising at least one additional agent, wherein the additional agent acts on the nervous system, or is an imaging agent, a cytotoxic agent, an angiogenesis inhibitor, a kinase inhibitor, a co-stimulation molecule blocker, an adhesion molecule blocker, an anti-cytokine antibody or functional fragment thereof, a detectable label or reporter, an antirheumatic, a muscle relaxant, a narcotic, a non-steroid anti-inflammatory drug (NSAID), an analgesic, an anesthetic, a sedative, a neuromuscular blocker, an antimicrobial, an antipsoriatic, a corticosteriod, an anabolic steroid, an erythropoietin, an immunoglobulin, an immunosuppressive, a growth hormone, a hormone replacement drug, a radiopharmaceutical, an antidepressant, an antipsychotic, a stimulant, an asthma medication, a beta agonist, an inhaled steroid, an epinephrine or analog, a cytokine, or a cytokine antagonist.

17. A method of detecting the presence, amount, or concentration of LRP-8 or a fragment of LRP-8 in a test sample by an immunoassay, comprising contacting the test sample with at least one binding protein and at least one detectable label, and wherein the at least one binding protein comprises the DVD-Ig binding protein of claim 1.

18. A dual variable domain immunoglobulin (DVD-Ig) binding protein that binds to human low density lipoprotein receptor-related protein 8 (LRP-8) at an epitope comprising amino acid residues 47-66 of human LRP-8 (SEQ ID NO:

1).

19. The DVD-Ig binding protein of claim 18, wherein the binding protein binds to human LRP-8 at an epitope comprising amino acid residues 47(C), 52(F), 56(N), 59(C), 60(I), 64(W), and 66(C) of human LRP-8 (SEQ ID NO:

1).

20. A monoclonal antibody that binds to human low density lipoprotein receptor-related protein 8 (LRP-8) at an epitope comprising amino acid residues 47-66 of human LRP-8 (SEQ ID NO:

1).

21. The antibody of claim 20, wherein the antibody binds to human LRP-8 at an epitope comprising amino acid residues 47(C), 52(F), 56(N), 59(C), 60(I), 64(W), and 66(C) of human LRP-8 (SEQ ID NO:

1).

22. A monoclonal antibody capable of binding low density lipoprotein receptor-related protein 8 (LRP-8), comprising a heavy chain variable domain comprising a set of three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2, and CDR-H3), and a light chain variable domain comprising a set of three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3), wherein the heavy chain CDRs are:

(a) CDR-H1;

RFTFSNX₁GMS (SEQ ID NO;

     9), wherein X₁ is F or Y, or GFTVSDYYMA (SEQ ID NO;

     53);

(b) CDR-H2;

TISSGGRYTYYPDX₂VKG (SEQ ID NO;

     10), wherein X₂ is S or H, or SISYEGSSTYYGDSVKG (SEQ ID NO;

     54); and

(c) CDR-H3;

DYLYAMDY (SEQ ID NO;

     46) or PLRYYGYNYRFAY (SEQ ID NO;

     55),

and the light chain CDRs are;

(d) CDR-L1;

RSSQSLVYSX₃X₄NTYLH (SEQ ID NO;

     47), wherein X₃ is N, T, R, W or P, and wherein X₄ is G, E, L or K, or KASQNIHKNLD (SEQ ID NO;

     56);

(e) CDR-L2;

KVSNRFS (SEQ ID NO;

     48) or YTDNLQT (SEQ ID NO;

     57); and

(f) CDR-L3;

SQSTHVPLT (SEQ ID NO;

     49) or YQYNSGPT (SEQ ID NO;

     58).

23. The antibody of claim 22, wherein the antibody comprises a heavy chain CDR set of:

(a) CDR-H1;

RFTFSNYGMS (SEQ ID NO;

     92);

(b) CDR-H2;

TISSGGRYTYYPDSVKG (SEQ ID NO;

     93); and

(c) CDR-H3;

DYLYAMDY (SEQ ID NO;

     46),

and a light chain CDR set of;

(d) CDR-L1;

RSSQSLVYSNGNTYLH (SEQ ID NO;

     157);

(e) CDR-L2;

KVSNRFS (SEQ ID NO;

     48); and

(f) CDR-L3;

SQSTHVPLT (SEQ ID NO;

     49).

24. The antibody of claim 22, wherein the antibody is a humanized antibody.

25. The antibody of claim 23, wherein the antibody is a humanized antibody.