IP Library › Granted Patent US 12,226,482
Granted Patent B2
US 12,226,482 · App. 18/454,286 · Granted Feb 18, 2025

Anti-SIRP-alpha antibodies and methods of use thereof

Inventors: Andrew Pincetic (San Francisco, CA); Arnon Rosenthal (Woodside, CA); Seung-Joo Lee (South San Francisco, CA)
Assignee: Alector LLC
A61K39/39558A61P31/00A61P35/00C07K16/2803C07K2317/24C07K2317/732C07K2317/75C07K2317/76C07K2317/77C07K2317/92
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Quick Facts
Patent No.
US 12,226,482
App. No.
18/454,286
Granted
Feb 18, 2025
Kind
B2
Abstract

The invention provides anti-SIRPA antibodies, methods of generating such antibodies, and therapeutic uses and methods employing the antibodies.

Claims (24)

1. An isolated anti-signal regulatory protein a (SIRPA) antibody, wherein the anti-SIRPA antibody comprises a heavy chain variable region (VH) that comprises a complementarity determining region 1 (CDR1) comprising the amino acid sequence of SEQ ID NO: 15, a CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 17; and a light chain variable region (VL) that comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a CDR2 comprising the amino acid sequence of SEQ ID NO: 13, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 14.

2. The anti-SIRPA antibody of claim 1 , wherein the antibody does not substantially block binding of CD47 to SIRPA expressed on cells, wherein binding of the antibody to SIRPA decreases the level of SIRPA expressed on the cell surface, and/or wherein the antibody binds to the D2 domain of SIRPA, or the D3 domain of SIRPA.

3. The anti-SIRPA antibody of claim 1 , wherein the antibody competes with an antibody comprising a VH sequence comprising the amino acid sequence of SEQ ID NO: 4 and a VL sequence comprising the amino acid sequence of SEQ ID NO: 5.

4. The anti-SIRPA antibody of claim 1 , wherein the VH comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of hSB-9C2-H1, hSB-9C2-H2, hSB-9C2-H3, or hSB-9C2-H4.

5. The anti-SIRPA antibody of claim 1 , wherein the VL comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of hSB-9C2-L1, hSB-9C2-L2, hsB-9C2-L3, or hSB-9C2-L4.

6. The anti-SIRPA antibody of claim 1 , wherein the VL comprises the amino acid sequence of hSB-9C2-L1, hSB-9C2-L2, hsB-9C2-L3, or hsB-9C2-L4.

7. The anti-SIRPA antibody of claim 1 , wherein the VH comprises the amino acid sequence of hSB-9C2-H1, hSB-9C2-H2, hSB-9C2-H3, or hSB-9C2-H4 and wherein the VL comprises the amino acid sequence of hSB-9C2-L1, hSB-9C2-L2, hsB-9C2-L3, or hsB-9C2-L4.

8. The anti-SIRPA antibody of claim 1 , wherein the VH comprises the amino acid sequence of SEQ ID NO: 4.

9. The anti-SIRPA antibody of claim 1 , wherein the VL comprises the amino acid sequence of SEQ ID NO: 5.

10. The anti-SIRPA antibody of claim 1 , wherein the VH region comprises the amino acid sequence of SEQ ID NO: 4 and the VL region comprises the amino acid sequence of SEQ ID NO: 5.

11. The anti-SIRPA antibody of claim 1 wherein the antibody is a humanized antibody.

12. The anti-SIRPA antibody of claim 1 , wherein the antibody is a Fab, Fab′, Fab′-SH, F (ab′)2, Fv or scFv fragment.

13. The anti-SIRPA antibody of claim 1 , wherein the anti-SIRPA antibody has an IgG1, IgG2, IgG3, or IgG4 isotype.

14. The anti-SIRPA antibody of claim 13 , wherein the anti-SIRPA antibody has a human IgG1 isotype and wherein the Fc region comprises substitution P331S; or substitutions L234A, L235A, and P331S; or substitutions S267E and L328F, wherein the numbering is according to EU numbering.

15. The anti-SIRPA antibody of claim 13 , wherein:

(a) the anti-SIRPA antibody has a human IgG1 isotype and comprises one or more amino acid substitutions in the Fc region at a residue position selected from the group consisting of: N297A, D265A, D270A, L234A, L235A, G237A, P238D, L328E, E233D, G237D, H268D, P271G, A330R, C226S, C229S, E233P, L234V, L234F, L235E, P331S, S267E, L328F, A330L, M252Y, S254T, T256E, N297Q, P238S, P238A, A327Q, A327G, P329A, K322A, T394D, and any combination thereof, or comprises an amino acid deletion in the Fc region at a position corresponding to glycine 236, wherein the numbering of the residues is according to EU numbering;

(b) the anti-SIRPA antibody has an IgG1 isotype and comprises an IgG2 isotype heavy chain constant domain 1 (CH1) and hinge region, optionally wherein the IgG2 isotype CH1 and hinge region comprises the amino acid sequence of ASTKGPSVFP LAPCSRSTSE STAALGCLVK DYFPEPVTVS WNSGALTSGVHTFPAVLQSS GLYSLSSVVT VPSSNFGTQT YTCNVDHKPS NTKVDKTVERKCCVECPPCP (SEQ ID NO:34), and optionally wherein the antibody Fc region comprises a S267E amino acid substitution, a L328F amino acid substitution, or both, and/or a N297A or N297Q amino acid substitution, wherein the numbering of the residues is according to EU numbering;

(c) the anti-SIRPA antibody has an IgG2 isotype and comprises one or more amino acid substitutions in the Fc region at a residue position selected from the group consisting of: P238S, V234A, G237A, H268A, H268Q, V309L, A330S, P331S, C214S, C232S, C233S, S267E, L328F, M252Y, S254T, T256E, H268E, N297A, N297Q, A330L, and any combination thereof, wherein the numbering of the residues is according to EU numbering;

(d) the anti-SIRPA antibody has a human IgG4 isotype and comprises one or more amino acid substitutions in the Fc region at a residue position selected from the group consisting of: L235A, G237A, S228P, L236E, S267E, E318A, L328F, M252Y, S254T, T256E, E233P, F234V, L234A/F234A, S228P, S241P, L248E, T394D, N297A, N297Q, L235E, and any combination thereof, wherein the numbering of the residues is according to EU numbering; or

(e) the anti-SIRPA antibody has a hybrid IgG2/4 isotype, and optionally wherein the antibody comprises an amino acid sequence comprising amino acids 118 to 260 of human IgG2 and amino acids 261 to 447 of human IgG4, wherein the numbering of the residues is according to EU numbering.

16. The anti-SIRPA antibody of claim 1 , wherein the VH comprises the amino acid sequence of hSB-9C2-H1, hSB-9C2-H2, hSB-9C2-H3, or hSB-9C2-H4.

17. The anti-SIRPA antibody of claim 1 , wherein the anti-SIRPA antibody is conjugated to a detectable marker, a toxin, or a therapeutic agent.

18. The anti-SIRPA antibody of claim 17 , wherein the anti-SIRPA antibody is conjugated to a toxin selected from the group consisting of ricin, ricin A chain, doxorubicin, daunorubicin, a maytansinoid, taxol, ethidium bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxy anthracin dione, actinomycin, diphtheria toxin, Pseudomonas exotoxin (PE) A, PE40, abrin, abrin A chain, modeccin A chain, alpha sarcin, gelonin, mitogellin, retstrictocin, phenomycin, enomycin, curicin, crotin, calicheamicin, Saponaria officinalis inhibitor, glucocorticoid, auristatin, auromycin, yttrium, bismuth, combrestatin, duocarmycins, dolastatin, cc1065, and a cisplatin.

19. A pharmaceutical composition comprising an anti-SIRPA antibody of claim 1 and a physiologically acceptable carrier.

Assignments (2)
SECURITY INTEREST Recorded Nov 14, 2024
From: ALECTOR, INC.; ALECTOR LLC
To: HERCULES CAPITAL, INC., AS AGENT
Reel/Frame 069386/0141 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 14, 2023
From: PINCETIC, ANDREW; ROSENTHAL, ARNON; LEE, SEUNG-JOO
To: ALECTOR LLC
Reel/Frame 065871/0469 →
Continuity (3)
Division 16463062
Provisional Application 62432503 · Dec 9, 2016
Related Publication 20240082395A1 · Mar 14, 2024
References Cited (105)
US 6180370B1 · Queen et al. · 2001 [cited by applicant]
US 8562997B2 · Jaiswal et al. · 2013 [cited by applicant]
US 9399682B2 · Jaiswal et al. · 2016 [cited by applicant]
US 9493575B2 · Jaiswal et al. · 2016 [cited by applicant]
US 9605076B2 · Jaiswal et al. · 2017 [cited by applicant]
US 9611329B2 · Jaiswal et al. · 2017 [cited by applicant]
US 9624305B2 · Jaiswal et al. · 2017 [cited by applicant]
US 9765143B2 · Jaiswal et al. · 2017 [cited by applicant]
US 10081680B2 · Weiskopf et al. · 2018 [cited by applicant]
US 20020090674A1 · Rosen et al. · 2002 [cited by applicant]
US 20020114807A1 · Berg et al. · 2002 [cited by applicant]
US 20030054415A1 · Buhring et al. · 2003 [cited by applicant]
US 20180105600A1 · Pons et al. · 2018 [cited by applicant]
US 20190359707A1 · Pincetic et al. · 2019 [cited by applicant]
CL 2019003266A1 · 2020 [cited by applicant]
CL 2020002305A1 · 2021 [cited by applicant]
CL 2021000738A1 · 2021 [cited by applicant]
CL 2021001260A1 · 2021 [cited by applicant]
CL 2021001627A1 · 2022 [cited by applicant]
CN 105026426A · 2015 [cited by applicant]
CN 107106679A · 2017 [cited by applicant]
WO 2001040307A1 · 2001 [cited by applicant]
WO 2002092784A2 · 2002 [cited by applicant]
WO 2009046541A1 · 2009 [cited by applicant]
WO 2009091547A1 · 2009 [cited by applicant]
WO 2009091601A1 · 2009 [cited by applicant]
WO 2009131453A1 · 2009 [cited by applicant]
WO 2010130053A1 · 2010 [cited by applicant]
WO 2013056352A1 · 2013 [cited by applicant]
WO 2014072876A1 · 2014 [cited by applicant]
WO 2014123580A1 · 2014 [cited by applicant]
WO 2014124028A1 · 2014 [cited by applicant]
WO 2015138600A2 · 2015 [cited by applicant]
WO 2015138600A3 · 2015 [cited by applicant]
WO 2015177360A1 · 2015 [cited by applicant]
WO 2016023019A2 · 2016 [cited by applicant]
WO 2016063233A1 · 2016 [cited by applicant]
WO 2016205042A1 · 2016 [cited by applicant]
WO 2017068164A1 · 2017 [cited by applicant]
WO 2017178653A2 · 2017 [cited by applicant]
WO 2017180519A1 · 2017 [cited by applicant]
WO 2017184553A1 · 2017 [cited by applicant]
WO 2017197495A1 · 2017 [cited by applicant]
WO 2018013534A1 · 2018 [cited by applicant]
WO 2018026600A1 · 2018 [cited by applicant]
WO 2018057669A1 · 2018 [cited by applicant]
WO 2018107058A1 · 2018 [cited by applicant]
WO 2017178653A3 · 2018 [cited by applicant]
WO 2018190719A2 · 2018 [cited by applicant]
WO 2018210795A1 · 2018 [cited by applicant]
WO 2018210793A3 · 2018 [cited by applicant]
WO 2019022600A1 · 2019 [cited by applicant]
WO 2019023347A1 · 2019 [cited by applicant]
WO 2019175218A1 · 2019 [cited by applicant]
WO 2020068752A1 · 2020 [cited by applicant]
WO 2020099653A1 · 2020 [cited by applicant]
WO 2020127373A1 · 2020 [cited by applicant]
Rudikoff, S. et al: “Single amino acid substitution altering antigen-binding specificity”, Proceedings of the National Academy of Sciences, National Academy of Sciences, vol. 79, Mar. 1, 1982 (Mar. 1, 1982), pp. 1979-19… [cited by applicant]
Ratnikova, et al., “CD47 receptor as a primary target for cancer therapy”, Mol Biol (Mosk), 51(2), 2017, 251-261 (11pgs.). [cited by applicant]
Ratnikova, et al., English language abstract (1 pg.), (2017). [cited by applicant]
Adams et al., “Signal-Regulatory Protein Is Selectively Expressed by Myeloid and Neuronal Cells,” J Immunol 1998; 161:1853-1859. [cited by applicant]
Ahmad, K., “Insulin sources and types: a review of insulin in terms of its mode on diabetes mellitus,” Journal of Traditional Chinese Medicine, vol. 34, issue 2 (2014). [cited by applicant]
Alblas et al., “Signal Regulatory Protein a Ligation Induces Macrophage Nitric Oxide Production through JAK/STAT- and Phosphatidylinositol 3-Kinase/Rac1/NAPDH Oxidase/H2O2-Dependent Pathways”, Molecular and Cellular Bio… [cited by applicant]
Anonymous, “MABS164 | Anti-SHPS-1 Antibody, clone P84,” 4 pages (Jan. 1, 2016). Retrieved from the Internet: http://www.merckmillipore.com/NL/en/product/Anti-SHPS-1-Antibody,-clone-P84,MM N F-MABS164#anchorPackaging%20I… [cited by applicant]
Barclay et al., “Signal regulatory protein alpha (SIRPα) / CD47 interaction and function,” Curr Opin Immunol. Feb. 2009 ; 21(1): pp. 47-52. [cited by applicant]
Berglund, L., et al., “The epitope space of the human proteome,” Protein Science, vol. 17, pp. 606-613 (2008). [cited by applicant]
Bio-Rad Antibodies, formerly AbD Serotec; Summary, (2021). [cited by applicant]
Bostrom et al., “Improving antibody binding affinity and specificity for therapeutic development”, Methods in Molecular Biology, 2009; vol. 525, pp. 353-376. [cited by applicant]
Chao et al., “Mechanisms of targeting CD47-SIRP in hematologic malignancies,” Blood, vol. 119, No. 18, pp. 4334-4336, May 3, 2012. [cited by applicant]
Chen, L., et al., “Epitope-directed antibody selection by site-specific photocrosslinking,” Science Advances, vol. 6, No. 14, p. eaaz7825 (2020). [cited by applicant]
Chiavenna et al., “State of the art in anti-cancer mAbs,” Journal of Biomedical Science (2017) 24:15. [cited by applicant]
Cosmetology, vol. 18, pp. 57-61 (2010). [cited by applicant]
De Vries et al., “Signal-regulatory protein alpha-CD47 interactions are required for the transmigration of monocytes across cerebral endothelium,” The Journal of Immunology, vol. 168, No. 11, pp. 5832-5839 (Jun. 1, 2002… [cited by applicant]
Edwards, B., et al., “The remarkable flexibility of the human antibody repertoire; isolation of over one thousand different antibodies to a single protein, BLyS,” Journal of Molecular Biology, vol. 334, pp. 103-118 (200… [cited by applicant]
Fournier et al., “Surfactant Protein D (Sp-D) Binds to Membrane-proximal Domain (D3) of Signal Regulatory Protein (SIRPα), a SiteDistant from Binding Domain of CD47, while Also Binding to Analogous Region on Signal Regu… [cited by applicant]
Gonzales, et al., “Minimizing the Immunogenicity of Antibodies for Clinical Application”, Tumour Biol., Jan.-Feb. 2005, vol. 26, No. 1, pp. 31-43. [cited by applicant]
Hatherly et al., “Polymorphisms in the Human Inhibitory Signal-regulatory Protein Do Not Affect Binding to Its Ligand CD47,” The Journal of Biological Chemistry, vol. 289, No. 14, pp. 10024-10028, Apr. 4, 2014. [cited by applicant]
Huang, R., et al., “Trastuzumab-cisplatin conjugates for targeted delivery of cisplatin to HER2-overexpressing cancer cells,” Biomedicine & Pharmacotherapy, vol. 72, pp. 17-23 (2015). [cited by applicant]
International Search Report and Written Opinion of International Application No. PCT/US2019/0363884, dated Aug. 27, 2019 (16 pages). [cited by applicant]
International Search Report and Written Opinion of PCT/US2017/065366 dated Mar. 26, 2018, (18 pages). [cited by applicant]
Janeway, C.A, Jr., et al., Immunobiology: The Immune System in Health and Disease, 5th edition, New York: Garland Science (2001). [cited by applicant]
Kipriyanov et al., “Generation and production of engineered antibodies,” Molecular Biotechnology, vol. 26, No. 1, pp. 39-60 (2004). [cited by applicant]
Kunik et al., “Structural consensus among antibodies defines the antigen binding site”, PLoS Comput Biol., Feb. 23, 2012, vol. 8, No. 2. [cited by applicant]
Lee et al., “Novel Structural Determinants on SIRPa that Mediate Binding to CD47,” Journal of Immunology, Jan. 1, 2007, vol. 179, No. 11, pp. 7741-7750. [cited by applicant]
Liu et al., “Functional Elements on SIRPα IgV domain Mediate Cell Surface Binding to CD47,” J Mol Biol. Jan. 19, 2007; 365(3): 680-693. [cited by applicant]
Liu et al., “Signal regulatory protein (SIRPα), a cellular ligand for CD47, regulates neutrophil transmigration,” The American Society for Biochemistry and Molecular Biology, J Biol Chem, 2002, 277:10028-10036. [cited by applicant]
Liu, Q., et al., “Inhibition of SIRP[alpha] in dendritic cells potentiates potent antitumor immunity,” Oncoimmunology, vol. 5, No. 9, p. e1183850 (2016). [cited by applicant]
Matozaki et al., “Functions and molecular mechanisms of the CD47-SIRPα signalling pathway,” Trends Cell Biol, 2009, 19:72-80. [cited by applicant]
Muchtar, E., et al., “Immunoglobulin Light-Chain Amyloidosis: From Basics to New Developments in Diagnosis, Prognosis and Therapy,” Acta Haematol, vol. 135, No. 3, pp. 172-190 (2016). [cited by applicant]
Murata et al., “The CD47SIRPa signalling system: its physiological roles and therapeutic application,” J. Biochem. 2014;155(6):335-344. [cited by applicant]
Oldenborg, “Cd47-Signal Regulatory Protein α (Sirpα) Regulates Fcγ and Complement Receptor-Mediated Phagocytosis,” The Journal of Experimental Medicine, vol. 193, No. 7, pp. 855-861, Apr. 2, 2001. [cited by applicant]
Padlan, E., “X-ray Crystallography of Antibodies”, Advances in Protein Chemistry, vol. 49, pp. 57-134 (1996). [cited by applicant]
Pan et al., “Signal Regulatory Protein a Is Associated With Tumor-Polarized Macrophages Phenotype Switch and Plays a Pivotal Role in Tumor Progression,” Hepatology, vol. 58, No. 2, pp. 680-691, 2013. [cited by applicant]
Rissiek, B., et al., “Nanobodies as modulators of inflammation: potential applications for acute brain injury,” Front Cell Neurosci, vol. 8, p. 344 (2014). [cited by applicant]
Seiffert et al., “Human Signal-Regulatory Protein Is Expressed on Normal, But Not on Subsets of Leukemic Myeloid Cells and MediatesCellular Adhesion Involving Its Counterreceptor CD47,” Blood, vol. 94, No. 11, pp. 3633-… [cited by applicant]
Seiffert et al., “Signal-regulatory protein a (SIRPα) but not SIRPβ is involved in T-Cell activation, binds to CD47 with high affinity, and is expressed on immature CD34+CD38-hematopoietic cells,” Blood, vol. 97, No. 9,… [cited by applicant]
Sharma, G., et al., “The Role of Cell-Penetrating Peptide and Transferrin on Enhanced Delivery of Drug to Brain,” International Journal of Molecular Sciences, vol. 17, No. 6, p. 806 (2016). [cited by applicant]
Sim, J., et al., “Discovery of high affinity, pan-allelic, and pan-mammalian reactive antibodies against the myeloid checkpoint receptors SIRP[alpha]”, mAbs, vol. 11, No. 6, pp. 1036-1052 (Jul. 1, 2019). [cited by applicant]
Van Beek et al., “Signal Regulatory Proteins in the Immune System,” J Immunol 2005; 175:7781-7787. [cited by applicant]
Wark et al., “Latest technologies for the enhancement of antibody affinity”, Advanced Drug Delivery Reviews, Elsevier, vol. 58, No. 5-6, Aug. 7, 2006, pp. 657-670. [cited by applicant]
Weiskopf et al.,“Cancer immunotherapy targeting the CD47/SIRPα axis”, Trends Cell Biol, 2009, 19:72-80. [cited by applicant]
Willingham et al., “The CD47-signal regulatory protein alpha (SIRPa) interaction is a therapeutic target for human solid tumors,” PNAS, vol. 109, No. 17, pp. 6662-6667, Apr. 24, 2012. [cited by applicant]
Yanagita et al., Anti-SIRPα antibodies as a potential new tool for cancer immunotherapy, JCI Insight, vol. 2, No. 1, pp. 2379-3708, Jan. 12, 2017. [cited by applicant]
Zhao et al., “CD47-signal regulatory protein-α (SIRPα) interactions form a barrier for antibody-mediated tumor cell destruction,” PNAS, vol. 108, No. 45, pp. 18342-18347, Nov. 8, 2011. [cited by applicant]
Zhao et al., “Is targeting of CD47-SIRP enough for treating hematopoietic malignancy?” Blood, vol. 119, No. 18, pp. 4333-4334, 2012. [cited by applicant]