Immunobinders directed against sclerostin
Proteins that bind sclerostin or sclerostin and TNF are described along with there use in composition and methods for treating, preventing, and diagnosing sclerostin related diseases and for detecting sclerostin or sclerostin and TNF in cells, tissues, samples, and compositions.
1. A binding protein comprising an antigen binding domain capable of binding human sclerostin, the antigen binding domain comprising three CDRs in a heavy chain and three CDRs in a light chain,
wherein the heavy chain comprises a CDR-H1, a CDR-H2 and a CDR-H3;
wherein the CDR-H1 comprises the amino acid sequence of X 1 -L-S-X 2 -H (residues 31-35 of SEQ ID NO: 1);
wherein X 1 is E or G; and
wherein X 2 is M, V or L;
wherein the CDR-H2 comprises the amino acid sequence of G-X 3 -X 4 -P-E-X 5 -G-E-X 6 -I-Y-A-Q-K-F-Q-G (residues 50-66 of SEQ ID NO: 1);
wherein X 3 is F or S;
wherein X 4 is D or N;
wherein X 5 is D, E, V, Y, N, G, A, R, Q, H, F or I; and
wherein X 6 is T, I, L, M or N; and
the CDR-H3 comprises the amino acid sequence of X 7 -X 8 -X 9 -X 10 -X 11 -W-X 12 -X 13 -X 14 -F-Q-H (residues 99-110 of SEQ ID NO: 1);
wherein X 7 is D or E;
wherein X 8 is S, T or A;
wherein X 9 is E, D, V, A, L, Y, W, Q, N, M, or G;
wherein X 10 is G or S;
wherein X 11 is Y, F, R, N, V, S, P or I;
wherein X 12 is E, Y, F or V;
wherein X 13 is K, Q or I; and
wherein X 14 is Y or F, and
wherein the light chain comprises a CDR-L1, a CDR-L2 and a CDR-L3;
wherein the CDR-L1 comprises an amino acid sequence of G-S-S-T-G-X 15 V-T-X 16 -X 17 -H-Y-X 18 -Y (residues 23-36 of SEQ ID NO: 2),
wherein X 15 is A, D, P, G, S, N E, T or H;
wherein X 16 is S, I, V, T, G or M;
wherein X 17 is G, E, D, W, Y or N; and
wherein X 18 is P or T;
wherein the CDR-L2 comprises the amino acid sequence of D-T-X 19 -X 20 -K-X 21 -S (residues 52-58 of SEQ ID NO: 2),
wherein X 19 is N, D, I, R, V, or T;
wherein X 20 is D or E; and
wherein X 21 is H, Q, N or D; and
wherein the CDR-L3 comprises the amino acid sequence of L-L-X 22 -X 23 -X 24 X -25 -X 26 -X 27 -V (residues 91-99 of SEQ ID NO: 2);
wherein X 22 is F or D;
wherein X 23 is Y, D, F, or N;
wherein X 24 is G or R;
wherein X 25 is G or S;
wherein X 26 is T, S, N or R; and
wherein X 27 is V, L, M or F.
2. The binding protein of claim 1 , wherein at least one CDR comprises an amino acid sequence selected from the group consisting of Residues 31-35 of SEQ ID NO:1998; Residues 50-66 of SEQ ID NO:1998; Residues 99-110 of SEQ ID NO:1998; Residues 31-35 of SEQ ID NO.:1999; Residues 50-66 of SEQ ID NO.:1999; Residues 99-110 of SEQ ID NO.:1999; Residues 31-35 of SEQ ID NO.:2000; Residues 50-66 of SEQ ID NO.:2000; Residues 99-110 of SEQ ID NO.:2000; Residues 31-35 of SEQ ID NO.:2001; Residues 50-66 of SEQ ID NO.:2001; Residues 99-110 of SEQ ID NO.:2001; Residues 31-35 of SEQ ID NO.:2002; Residues 50-66 of SEQ ID NO.:2002; Residues 99-110 of SEQ ID NO.:2002; Residues 31-35 of SEQ ID NO.:2003; Residues 50-66 of SEQ ID NO.:2003; Residues 99-110 of SEQ ID NO.:2003; Residues 31-35 of SEQ ID NO.:2004; Residues 50-66 of SEQ ID NO.:2004; Residues 99-110 of SEQ ID NO.:2004; Residues 31-35 of SEQ ID NO.:2005; Residues 50-66 of SEQ ID NO.:2005; Residues 99-110 of SEQ ID NO.:2005; Residues 31-35 of SEQ ID NO.:2006; Residues 50-66 of SEQ ID NO.:2006; Residues 99-110 of SEQ ID NO.:2006; Residues 31-35 of SEQ ID NO.:2007; Residues 50-66 of SEQ ID NO.:2007; Residues 99-110 of SEQ ID NO.:2007; Residues 23-36 of SEQ ID NO.:2008; Residues 52-58 of SEQ ID NO.:2008; Residues 91-99 of SEQ ID NO.:2008; Residues 23-36 of SEQ ID NO.:2009; Residues 52-58 of SEQ ID NO.:2009; Residues 91-99 of SEQ ID NO.:2009; Residues 23-36 of SEQ ID NO.:2010; Residues 52-58 of SEQ ID NO.:2010; Residues 91-99 of SEQ ID NO.:2010; Residues 23-36 of SEQ ID NO.:2011; Residues 52-58 of SEQ ID NO.:2011; Residues 91-99 of SEQ ID NO.:2011; Residues 23-36 of SEQ ID NO.:2012; Residues 52-58 of SEQ ID NO.:2012; Residues 91-99 of SEQ ID NO.:2012; Residues 23-36 of SEQ ID NO.:2013; Residues 52-58 of SEQ ID NO.:2013; Residues 91-99 of SEQ ID NO.:2013; Residues 23-36 of SEQ ID NO.:2014; Residues 52-58 of SEQ ID NO.:2014; Residues 91-99 of SEQ ID NO.:2014; Residues 23-36 of SEQ ID NO.:2015; Residues 52-58 of SEQ ID NO.:2015; Residues 91-99 of SEQ ID NO.:2015; Residues 23-36 of SEQ ID NO.:2016; Residues 52-58 of SEQ ID NO.:2016; Residues 91-99 of SEQ ID NO.:2016; Residues 23-36 of SEQ ID NO.:2017; Residues 52-58 of SEQ ID NO.:2017; Residues 91-99 of SEQ ID NO.:2017.
3. The binding protein of claim 1 , wherein the antigen binding domain comprises a variable domain CDR set selected from the group consisting of:
VH AE10-6 AM1 CDR Set
VH AE10-6 AM1 CDR-H1
Residues 31-35 of SEQ ID NO: 1998
VH AE10-6 AM1 CDR-H2
Residues 50-66 of SEQ ID NO: 1998
VH AE10-6 AM1 CDR-H3
Residue 99-110 of SEQ ID NO: 1998
VL AE10-6 AM1 Set
VL AE10-6 AM1 CDR-L1
Residues 23-36 of SEQ ID NO: 2008
VL AE10-6 AM1 CDR-L2
Residues 52-58 of SEQ ID NO: 2008
VL AE10-6 AM1 CDR-L3
Residues 91-99 of SEQ ID NO: 2008
VH AE10-6 AM2 CDR Set
VH AE10-6 AM2 CDR-H1
Residues 31-35 of SEQ ID NO: 1999
VH AE10-6 AM2 CDR-H2
Residues 50-66 of SEQ ID NO: 1999
VH AE10-6 AM2 CDR-H3
Residue 99-110 of SEQ ID NO: 1999
VL AE10-6 AM2 Set
VL AE10-6 AM2 CDR-L1
Residues 23-36 of SEQ ID NO: 2009
VL AE10-6 AM2 CDR-L2
Residues 52-58 of SEQ ID NO: 2009
VL AE10-6 AM2 CDR-L3
Residues 91-99 of SEQ ID NO: 2009
VH AE10-6 AM3 CDR Set
VH AE10-6 AM3 CDR-H1
Residues 31-35 of SEQ ID NO: 2000
VH AE10-6 AM3 CDR-H2
Residues 50-66 of SEQ ID NO: 2000
VH AE10-6 AM3 CDR-H3
Residue 99-110 of SEQ ID NO: 2000
VL AE10-6 AM3 Set
VL AE10-6 AM3 CDR-L1
Residues 23-36 of SEQ ID NO: 2010
VL AE10-6 AM3 CDR-L2
Residues 52-58 of SEQ ID NO: 2010
VL AE10-6 AM3 CDR-L3
Residues 91-99 of SEQ ID NO: 2010
VH AE10-6 AM4 CDR Set
VH AE10-6 AM4 CDR-H1
Residues 31-35 of SEQ ID NO: 2001
VH AE10-6 AM4 CDR-H2
Residues 50-66 of SEQ ID NO: 2001
VH AE10-6 AM4 CDR-H3
Residue 99-110 of SEQ ID NO: 2001
VL AE10-6 AM4 Set
VL AE10-6 AM4 CDR-L1
Residues 23-36 of SEQ ID NO: 2011
VL AE10-6 AM4 CDR-L2
Residues 52-58 of SEQ ID NO: 2011
VL AE10-6 AM4 CDR-L3
Residues 91-99 of SEQ ID NO: 2011
VH AE10-6 AM5 CDR Set
VH AE10-6 AM5 CDR-H1
Residues 31-35 of SEQ ID NO: 2002
VH AE10-6 AM5 CDR-H2
Residues 50-66 of SEQ ID NO: 2002
VH AE10-6 AM5 CDR-H3
Residue 99-110 of SEQ ID NO: 2002
VL AE10-6 AM5 Set
VL AE10-6 AM5 CDR-L1
Residues 23-36 of SEQ ID NO: 2012
VL AE10-6 AM5 CDR-L2
Residues 52-58 of SEQ ID NO: 2012
VL AE10-6 AM5 CDR-L3
Residues 91-99 of SEQ ID NO: 2012
VH AE10-6 AM6 CDR Set
VH AE10-6 AM6 CDR-H1
Residues 31-35 of SEQ ID NO: 2003
VH AE10-6 AM6 CDR-H2
Residues 50-66 of SEQ ID NO: 2003
VH AE10-6 AM6 CDR-H3
Residue 99-110 of SEQ ID NO: 2003
VL AE10-6 AM6 Set
VL AE10-6 AM6 CDR-L1
Residues 23-36 of SEQ ID NO: 2013
VL AE10-6 AM6 CDR-L2
Residues 52-58 of SEQ ID NO: 2013
VL AE10-6 AM6 CDR-L3
Residues 91-99 of SEQ ID NO: 2013
VH AE10-6 AM7 CDR Set
VH AE10-6 AM7 CDR-H1
Residues 31-35 of SEQ ID NO: 2004
VH AE10-6 AM7 CDR-H2
Residues 50-66 of SEQ ID NO: 2004
VH AE10-6 AM7 CDR-H3
Residue 99-110 of SEQ ID NO: 2004
VL AE10-6 AM7 Set
VL AE10-6 AM7 CDR-L1
Residues 23-36 of SEQ ID NO: 2014
VL AE10-6 AM7 CDR-L2
Residues 52-58 of SEQ ID NO: 2014
VL AE10-6 AM7 CDR-L3
Residues 91-99 of SEQ ID NO: 2014
VH AE10-6 AM8 CDR Set
VH AE10-6 AM8 CDR-H1
Residues 31-35 of SEQ ID NO: 2005
VH AE10-6 AM8 CDR-H2
Residues 50-66 of SEQ ID NO: 2005
VH AE10-6 AM8 CDR-H3
Residue 99-110 of SEQ ID NO: 2005
VL AE10-6 AM8 Set
VL AE10-6 AM8 CDR-L1
Residues 23-36 of SEQ ID NO: 2015
VL AE10-6 AM8 CDR-L2
Residues 52-58 of SEQ ID NO: 2015
VL AE10-6 AM8 CDR-L3
Residues 91-99 of SEQ ID NO: 2015
VH AE10-6 AM9 CDR Set
VH AE10-6 AM9 CDR-H1
Residues 31-35 of SEQ ID NO: 2006
VH AE10-6 AM9 CDR-H2
Residues 50-66 of SEQ ID NO: 2006
VH AE10-6 AM9 CDR-H3
Residue 99-110 of SEQ ID NO: 2006
VL AE10-6 AM9 Set
VL AE10-6 AM9 CDR-L1
Residues 23-36 of SEQ ID NO: 2016
VL AE10-6 AM9 CDR-L2
Residues 52-58 of SEQ ID NO: 2016
VL AE10-6 AM9 CDR-L3
Residues 91-99 of SEQ ID NO: 2016
VH AE10-6 AM10 CDR Set
VH AE10-6 AM10 CDR-H1
Residues 31-35 of SEQ ID NO: 2007
VH AE10-6 AM10 CDR-H2
Residues 50-66 of SEQ ID NO: 2007
VH AE10-6 AM10 CDR-H3
Residue 99-110 of SEQ ID NO: 2007
VL AE10-6 AM10 Set
VL AE10-6 AM10 CDR-L1
Residues 23-36 of SEQ ID NO: 2017
VL AE10-6 AM10 CDR-L2
Residues 52-58 of SEQ ID NO: 2017
VL AE10-6 AM10 CDR-L3
Residues 91-99 of SEQ ID NO: 2017.
4. The binding protein of claim 3 , comprising at least two variable domain CDR sets.
5. The binding protein of claim 4 , wherein the at least two variable domain CDR sets are selected from the group consisting of VH AE10-6 AM1 CDR Set and VL AE10-6 AM1 CDR Set; VH AE10-6 AM2 CDR Set and VL AE10-6 AM2 CDR Set; VH AE10-6 AM3 CDR Set and VL AE10-6 AM3 CDR Set; VH AE10-6 AM4 CDR Set and VL AE10-6 AM4 CDR Set; VH AE10-6 AM5 CDR Set and VL AE10-6 AM5CDR Set; VH AE10-6 AM6 CDR Set and VL AE10-6 AM6 CDR Set; VH AE10-6 AM7 CDR Set and VL AE10-6 AM7 CDR Set; VH AE10-6 AM8 CDR Set and VL AE10-6AM8_CDR Set; VH AE10-6 AM9_CDR Set and VL AE10-6 AM9CDR Set; VH AE10-6 AM10_CDR Set and VL AE10-6 AM10 CDR Set.
6. The binding protein of claim 1 , wherein the binding protein comprises at least one variable domain having an amino acid sequence selected from the group consisting of SEQ ID NOs 1719-1866; SEQ ID NOs1867-1997; SEQ ID NOs 1998-2007; and SEQ ID NOs 2008-2017.
7. The binding protein of claim 1 , wherein the binding protein comprises two variable domains, wherein the first variable domain comprises a sequence selected from the group consisting of SEQ ID NOs 1719-1866, and 1998-2007 and wherein the second variable domain comprises a sequence selected from the group consisting of SEQ ID NOs 1867-1997, and 2007-2017.
8. The binding protein of claim 1 , wherein the binding protein:
(a) is capable of modulating a biological function of sclerostin;
(b) is capable of neutralizing sclerostin;
(c) wherein the sclerostin is pro-human sclerostin; mature-human sclerostin, or truncated-human sclerostin;
(d) wherein the binding protein diminishes the ability of sclerostin to bind to its receptor; and/or
(e) wherein the neutralizing binding protein is capable of reducing one or more of Th1 modulation; Th2 modulation; Nk modulation; neutrophil modulation; monocyte-macrophage lineage modulation; neutrophil modulation; eosinophil modulation; B-cells modulation; cytokine modulation; chemokine modulation; adhesion molecule modulation; and cell recruitment modulation.
9. A pharmaceutical composition comprising the binding protein of claim 1 , and a pharmaceutically acceptable carrier.
10. The pharmaceutical composition of claim 9 , further comprising at least one additional therapeutic agent for treating a disorder in which sclerostin activity is detrimental.
11. The pharmaceutical composition of claim 10 , wherein the additional agent is selected from the group consisting of a therapeutic agent, imaging agent, cytotoxic agent, angiogenesis inhibitors; kinase inhibitors; co-stimulation molecule blockers; adhesion molecule blockers; anti-cytokine antibody or functional fragment thereof; methotrexate; cyclosporin; rapamycin; FK506; detectable label or reporter; a TNF antagonist; an anti-rheumatic; a muscle relaxant, a narcotic, a non-steroid anti-inflammatory drug (NSAID), an analgesic, an anesthetic, a sedative, a local anesthetic, a neuromuscular blocker, an antimicrobial, an antipsoriatic, a corticosteroid, an anabolic steroid, an erythropoietin, an immunization, 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 oral steroid, an epinephrine or analog, a cytokine, and a cytokine antagonist.
12. The binding protein of claim 1 , wherein the antigen binding domain comprises a heavy chain variable region (VH) comprising three CDRs of SEQ ID NO:2004.
13. The binding protein of claim 1 , wherein the antigen binding domain comprises a light chain variable region (VL) comprising three CDRs of SEQ ID NO:2014.
14. The binding protein of claim 1 , wherein the antigen binding domain comprises a heavy chain variable region (VH) comprising three CDRs from SEQ ID NO:2004; and wherein the antigen binding domain comprises a light chain variable region (VL) comprising three CDRs of SEQ ID NO:2014.
15. The binding protein of claim 1 , wherein the antigen binding domain comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:2004.
16. The binding protein of claim 1 , wherein the antigen binding domain comprises a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:2014.
17. The binding protein of claim 1 , wherein the antigen binding domain comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:2004; and wherein the antigen binding domain comprises a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:2014.