SIRP alpha fusion protein formulation

Abstract

The present invention relates to the field of pharmaceutical formulations of proteins. Specifically, the present invention relates to a stable liquid SIRPαFc formulation and its pharmaceutical preparation and use.

Description

[0001] PC073180A

[0002] SIRP ALPHA FUSION PROTEIN FORMULATION

[0003] Reference to Sequence Listing

[0004] The instant application contains a Sequence Listing which has been submitted electronically in .xml format and is hereby incorporated by reference in its entirety. Said Sequence Listing, created on September 20, 2024, is named “PC073180_SeqListing_ST26.xml” and is 14 kilobytes in size.

[0005] Background

[0006] Cancer cells are targeted for destruction by antibodies that bind to cancer cell antigens, and through recruitment and activation of macrophages by way of Fc receptor binding to the Fc portion of that antibody. Binding between CD47 on cancer cells and SIRPa on macrophages transmits a “don’t eat me” signal that enables many tumour cells to escape destruction by macrophages. It has been shown that inhibition of the CD47 / SIRPa interaction (CD47 blockade) will allow macrophages to “see” and destroy the target CD47+ cancer cell. The use of SIRPa to treat cancer by CD47 blockade is described in WO 2010 / 130053, incorporated herein by reference. International Patent Application Publication No. WO 2014 / 094122, incorporated by reference in its entirety, describes a protein drug that inhibits the interaction between CD47 and SIRPa. This CD47 blockade drug is a form of human SIRPa that incorporates a particular region of its extracellular domain linked with a particularly useful form of an IgG-based Fc region. In this form, the SIRPaFc fusion drug shows dramatic effects on the viability of cancer cells that present with a CD47+ phenotype.

[0007] Protein preparations intended for therapeutic or prophylactic use require stabilizers to prevent loss of activity or structural integrity of the protein due to the effects of denaturation, oxidation or aggregation over a period of time during storage and transportation prior to use. These problems are exacerbated at the high concentrations of protein often desired for therapeutic administration.

[0008] A major aim in the development of protein formulations is to maintain protein solubility, stability and potency. It is particularly desirable to avoid aggregates and particulates in solution which would require sterile filtration before use for intravenous or subcutaneous injection and limit route of administration. Formulation of protein preparations requires careful selection of these factors among others to avoid denaturation of the protein and loss of protein-binding activity. Accordingly, there is a need for a stable aqueous protein formulation which stably supports high concentrations of bioactive protein in solution and is suitable for parenteral administration, including intravenous, intraocular, intravitreal, intraarterial, intrathecal, intraventricular, intraurethral, intrasternal, intracranial, intramuscular, intra-ossial, intraperitoneal, intradermal or subcutaneous injection.

[0009] Furthermore, there is a need to provide such a stable aqueous formulation for a SIRPaFc fusion protein. There is a need for a stable aqueous preparation of an SIRPaFc fusion protein to meet the medical need of patients suffering from cancer.

[0010] Summary

[0011] Stable aqueous pharmaceutical formulations with an extended shelf life comprising a SIRPalpha (SIRPa) Fc fusion protein (SIRPaFc) are provided. It is demonstrated that the aqueous pharmaceutical formulation of the present invention with high protein concentration is stable (e.g., having low levels of % HMMS (High Molecular Mass Species) and high levels of % soluble SIRPaFc) and suitable for parenteral administration.

[0012] In some embodiments, provided herein is an aqueous formulation comprising: 10 mg / mL to 300 mg / mL of a SIRPalpha-Fc fusion protein (SIRPaFc), a buffer, a sugar, a surfactant, and a chelating agent, wherein the formulation has a pH of 5.5 to 6.5.

[0013] In some embodiments, provided herein is an aqueous formulation comprising: 10 mg / mL to 300 mg / mL of a SIRPaFc; 5-50 mM of a histidine buffer; 1 mg / ml to 300 mg / ml of sucrose; 0.01 mg / ml to 10 mg / ml of PS80; 0.01 mg / ml to 1 mg / ml of EDTA; and wherein the formulation has a pH of 5.5 to 6.5.

[0014] In some embodiments, provided herein is an aqueous formulation comprising: 150 mg / ml of a SIRPaFc; 20 mM histidine; 60 mg / ml sucrose; 0.2 mg / ml PS80; 0.05 mg / ml EDTA; wherein the SIRPaFc comprises the amino acid sequence as shown in SEQ ID NO: 7 and wherein the formulation has a pH of 6.

[0015] In some embodiments, provided herein is a method for treating or inhibiting cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the aqueous formulation as provided herein.

[0016] In some embodiments, provided herein is the aqueous formulation provided herein for use as a medicament. In some embodiments, provided herein is the aqueous formulation provided herein for use in a method for treating or inhibiting cancer in a subject. In some embodiments, provided herein is the use of the aqueous formulation provided herein for the manufacture of a medicament for treatment of cancer in a subject.

[0017] Brief Description of the Figures / Drawings

[0018] FIG. 1A is a table summarizing % soluble maplirpacept molecules (“mono”) in formulation samples after storage at 25°C for 0 or 2 weeks (“W”) at pH 4, 5, 6, 7, 7.4, or 8. FIG. 1 B is a table summarizing % soluble maplirpacept molecules (“mono”) in samples after storage at 40°C for 0, 1 or 2 weeks (“W”) at pH 4, 5, 6, 7, 7.4, or 8.

[0019] FIG. 2A is a graph showing % acidic species in maplirpacept formulations after storage at 25°C for 0 or 2 weeks (“W”) at pH 4, 5, 6, 7, 7.4, or 8. FIG. 2B is a graph showing % acidic species in maplirpacept formulations after storage at 40°C for 0, 1 or 2 weeks (“W”) at pH 4, 5, 6, 7, 7.4, or 8.

[0020] Detailed Description

[0021] Disclosed herein are stable aqueous pharmaceutical formulations with an extended shelf-life comprising a SIRPalpha Fc fusion protein (SIRPaFc). It is demonstrated that the aqueous pharmaceutical formulation of the present invention stably supports high concentration of SIRPaFc (e.g., having low levels of % HMMS (High Molecular Mass Species) at a SIRPaFc concentration of at least about 15 mg / mL) and is suitable for parenteral administration, including subcutaneous, intravenous, intramuscular, intraperitoneal, or intradermal injection. Accordingly, in one aspect, provided is an aqueous formulation comprising: about 10 mg / ml to about 300 mg / ml of a SIRPaFc fusion protein; a buffer; a sugar; a surfactant; a chelating agent; and wherein the formulation has a pH at about 5.5 to about 6.5. For example, in some embodiments, provided is an aqueous formulation comprising: about 10 mg / ml to about 300 mg / ml of a SIRPaFc fusion protein; about 5 mM to about 50 mM of a buffer (e.g., histidine buffer); about 1 mg / mL to about 300 mg / mL of a sugar (e.g., sucrose); about 0.01 mg / ml to about 10 mg / ml of a surfactant (e.g., polysorbate 80); about 0.01 mg / ml to about 10.0 mg / ml of a chelating agent (e.g., EDTA (or edetate)); wherein the formulation has a pH at about 5.5 to about 6.5. In some embodiments, the SIRPaFc fusion protein concentration is 150 mg / mL or 200 mg / mL. Exemplary embodiments (E) of the invention provided herein include:

[0022] E1 . An aqueous formulation comprising: 10 mg / mL to 300 mg / mL of a SIRPalpha- Fc fusion protein (SIRPaFc), a buffer, a sugar, a surfactant, and a chelating agent, wherein the formulation has a pH of 5.5 to 6.5.

[0023] E2. The aqueous formulation of E1 , wherein the buffer is a histidine buffer.

[0024] E3. The aqueous formulation of any one of E1-E2, wherein the concentration of the buffer is 5-50 mM.

[0025] E4. The aqueous formulation of any one of E1-E3, wherein the sugar is sucrose.

[0026] E5. The aqueous formulation of any one of E1-E4, wherein the concentration of the sugar is about 1 mg / ml to about 300 mg / ml.

[0027] E6. The aqueous formulation of any one of E1-E5, wherein the surfactant is a polysorbate.

[0028] E7. The aqueous formulation of E6, wherein the polysorbate is polysorbate 80 (PS80).

[0029] E8. The aqueous formulation of E7, wherein the concentration of the surfactant is 0.01 mg / ml to 10 mg / ml.

[0030] E9. The aqueous formulation of any one of E1 to E8, wherein the chelating agent is ethylenediaminetetracetic acid (EDTA).

[0031] E10. The aqueous formulation of any one of E1 to E9, wherein the concentration of the chelating agent is about 0.01 mg / ml to about 1.0 mg / ml.

[0032] E11. The aqueous formulation of any one of E1 -E10, wherein the SIRPa portion of the SIRPaFc comprises the amino acid sequence shown in SEQ ID NO: 2.

[0033] E12. The aqueous formulation of any one of E1-E11 , wherein the SIRPaFc comprises the amino acid sequence as shown in SEQ ID NO: 6 or SEQ ID NO: 7. E13. The aqueous formulation of any one of E1-E12, wherein the formulation has a shelf life of at least 12 weeks.

[0034] E14. An aqueous formulation comprising: 10 mg / mL to 300 mg / mL of a SIRPaFc; 5-50 mM of a histidine buffer; 1 mg / ml to 300 mg / ml of sucrose; 0.01 mg / ml to 10 mg / ml of PS80; 0.01 mg / ml to 1 mg / ml of EDTA; and wherein the formulation has a pH of 5.5 to 6.5.

[0035] E15. The aqueous formulation of E14, wherein the SIRPaFc comprises the amino acid sequence as shown in SEQ ID NO: 6 or SEQ ID NO: 7. E16. The aqueous formulation of any one of E1-E15, wherein the SIRPaFc is at a concentration of 50 mg / ml, 75 mg / ml, 100 mg / ml, 125 mg / ml, 150 mg / ml, 175 mg / ml, or 200 mg / ml.

[0036] E17. An aqueous formulation comprising: 150 mg / ml of a SIRPaFc; 20 mM histidine; 60 mg / ml sucrose; 0.2 mg / ml PS80; 0.05 mg / ml EDTA; wherein the SIRPaFc comprises the amino acid sequence as shown in SEQ ID NO: 7 and wherein the formulation has a pH of 6.

[0037] E18. The aqueous formulation of any one of E1 -E17, wherein the formulation has a shelf life of at least 12 weeks.

[0038] E19. A method for treating or inhibiting cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the aqueous formulation of any one of E1 to E18.

[0039] E20. The method of E19, wherein the cancer is a hematological cancer.

[0040] E21. The method of E19 or E20, wherein the formulation is administered to the subject subcutaneously or intravenously.

[0041] E22. The aqueous formulation according to any of E1 to E18, for use in a method for treating or inhibiting cancer in a subject.

[0042] E23. The aqueous formulation for use of E22, wherein the cancer is a hematological cancer.

[0043] E24. The aqueous formulation for use of E22 or E23, wherein the formulation is administered to the subject subcutaneously or intravenously.

[0044] E25. Use of the aqueous formulation according to any of E1 to E18 for the manufacture of a medicament for treatment of cancer in a subject.

[0045] E26. The use of E25, wherein the cancer is a hematological cancer.

[0046] E27. The use of E25 or E26, wherein the formulation is administered to the subject subcutaneously or intravenously.

[0047] E28. The method of any one of E19 to E21 , aqueous formulation for use of any one of E22 to E24, or the use of any one of E25 to E27, wherein the subject is human.

[0048] General Techniques

[0049] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, which are within the skill of the art. Such techniques are explained fully in the literature, such as, Molecular Cloning: A Laboratory Manual, second edition (Sambrook et al., 1989) Cold Spring Harbor Press; Oligonucleotide Synthesis (M.J. Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (J.E. Cellis, ed., 1998) Academic Press; Animal Cell Culture (R. l. Freshney, ed., 1987); Introduction to Cell and Tissue Culture (J.P. Mather and P.E. Roberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (A. Doyle, J.B. Griffiths, and D.G. Newell, eds., 1993-1998) J. Wiley and Sons; Methods in Enzymology (Academic Press, Inc.); Handbook of Experimental Immunology (D.M. Weir and C.C. Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (J.M. Miller and M.P. Calos, eds., 1987); Current Protocols in Molecular Biology (F.M. Ausubel et al., eds., 1987); PCR: The Polymerase Chain Reaction, (Mullis et al., eds., 1994); Current Protocols in Immunology (J.E. Coligan et al., eds., 1991 ); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (C.A. Janeway and P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: a practical approach (D. Catty., ed., IRL Press, 1988-1989); Monoclonal antibodies: a practical approach (P. Shepherd and C. Dean, eds., Oxford University Press, 2000); Using antibodies: a laboratory manual (E. Harlow and D. Lane (Cold Spring Harbor Laboratory Press, 1999); The Antibodies (M. Zanetti and J.D. Capra, eds., Harwood Academic Publishers, 1995), as well as in subsequent editions and corresponding websites of the above references, as applicable. Definitions

[0050] Unless otherwise defined, all terms of art, notations and other scientific terms or terminology used herein are intended to have the meanings commonly understood by those of skill in the art to which this invention pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art.

[0051] The following terms, unless otherwise indicated, shall be understood to have the following meanings:

[0052] The term "isolated molecule" (where the molecule is, for example, a polypeptide, a polynucleotide, or an antibody) is a molecule that by virtue of its origin or source of derivation (1 ) is not associated with naturally associated components that accompany it in its native state, (2) is substantially free of other molecules from the same species (3) is expressed by a cell from a different species, or (4) does not occur in nature. Thus, a molecule that is chemically synthesized, or expressed in a cellular system different from the cell from which it naturally originates, will be "isolated" from its naturally associated components. A molecule also may be rendered substantially free of naturally associated components by isolation, using purification techniques well known in the art. Molecule purity or homogeneity may be assayed by a number of means well known in the art. For example, the purity of a polypeptide sample may be assayed using polyacrylamide gel electrophoresis and staining of the gel to visualize the polypeptide using techniques well known in the art. For certain purposes, higher resolution may be provided by using HPLC or other means well known in the art for purification.

[0053] As used herein, the term "formulation" as it relates to an antibody is meant to describe the antibody in combination with a pharmaceutically acceptable excipient comprising at least one buffer, at least one surfactant, at least one chelating agent, and wherein the pH is as defined.

[0054] The terms "pharmaceutical composition" or “pharmaceutical formulation” refer to preparations which are in such form as to permit the biological activity of the active ingredients to be effective.

[0055] "Pharmaceutically acceptable excipients" (vehicles, additives) are those, which can safely be administered to a subject to provide an effective dose of the active ingredient employed. The term "excipient" or "carrier" as used herein refers to an inert substance, which is commonly used as a diluent, vehicle, preservative, binder or stabilizing agent for drugs. As used herein, the term "diluent" refers to a pharmaceutically acceptable (safe and non-toxic for administration to a human) solvent and is useful for the preparation of the aqueous formulations herein. Exemplary diluents include, but are not limited to, sterile water and bacteriostatic water for injection (BWFI).

[0056] A "therapeutically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result, which in the context of SIRPaFc fusion protein includes treatment or prophylactic prevention of the targeted pathologic condition for example cancer. It is to be noted that dosage values may vary with the seventy of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that dosage ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition. Likewise, a therapeutically effective amount of the antibody or antibody portion may vary according to factors such as the disease state, age, sex, and weight of the individual, the ability of the antibody or antibody portion to elicit a desired response in the individual, and the desired route of administration of the antibody formulation. A therapeutically effective amount is also one in which any toxic or detrimental effects of the antibody or antibody portion are outweighed by the therapeutically beneficial effects.

[0057] As used herein, the term "treatment" refers to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) the targeted pathologic condition (e.g., any condition that would benefit from treatment with the antibody. This includes chronic and acute disorders or diseases including those pathological conditions which predispose the mammal to the disorder in question. Non-limiting examples of disorders to be treated herein include benign and malignant tumors; leukemias and lymphoid malignancies; neuronal, glial, astrocytal, hypothalamic and other glandular, macrophagal, epithelial, stromal and blastocoelic disorders; and inflammatory, angiogenic and immunologic disorders.). Those in need of treatment include those already with the condition as well as those prone to have the condition or those in whom the condition is to be prevented. As used herein, “treatment” is an approach for obtaining beneficial or desired clinical results including, but not limited to, one or more of the following: including lessening seventy, alleviation of one or more symptoms associated with the pathologic condition.

[0058] An “effective amount” of drug, formulation, compound, or pharmaceutical composition is an amount sufficient to effect beneficial or desired results including clinical results such as alleviation or reduction of the targeted pathologic condition. An effective amount can be administered in one or more administrations. For purposes of this invention, an effective amount of drug, compound, or pharmaceutical composition is an amount sufficient to treat, ameliorate, or reduce the intensity of the targeted pathologic condition. As is understood in the clinical context, an effective amount of a drug, compound, or pharmaceutical composition may or may not be achieved in conjunction with another drug, compound, or pharmaceutical composition. Thus, an “effective amount” may be considered in the context of administering one or more therapeutic agents, and a single agent may be considered to be given in an effective amount if, in conjunction with one or more other agents, a desirable result may be or is achieved.

[0059] As used herein, the term "subject" for purposes of treatment includes any subject, and preferably is a subject who is in need of the treatment of the targeted pathologic condition (e.g., cancer). For purposes of prevention, the subject is any subject, and preferably is a subject that is at risk for, or is predisposed to, developing the targeted pathologic condition. The term "subject" is intended to include living organisms, e.g., prokaryotes and eukaryotes. Examples of subjects include mammals, e.g., humans, dogs, cows, horses, pigs, sheep, goats, cats, mice, rabbits, rats, and transgenic non- human animals. In specific embodiments of the invention, the subject is a human.

[0060] As used herein, the term "polynucleotide" or "nucleic acid", used interchangeably herein, means a polymeric form of nucleotides either ribonucleotides or deoxynucleotides or a modified form of either type of nucleotide and may be single and double stranded forms. A "polynucleotide" or a "nucleic acid" sequence encompasses its complement unless otherwise specified. As used herein, the term "isolated polynucleotide" or "isolated nucleic acid" means a polynucleotide of genomic, cDNA, or synthetic origin or some combination thereof, which by virtue of its origin or source of derivation, the isolated polynucleotide has one to three of the following: (1 ) is not associated with all or a portion of a polynucleotide with which the "isolated polynucleotide" is found in nature, (2) is operably linked to a polynucleotide to which it is not linked in nature, or (3) does not occur in nature as part of a larger sequence.

[0061] As used herein, "pharmaceutically acceptable carrier" includes any material which, when combined with an active ingredient, allows the ingredient to retain biological activity and is non-reactive with the subject's immune system. Examples include, but are not limited to, any of the standard pharmaceutical carriers such as a phosphate buffered saline solution, water, emulsions such as oil / water emulsion, and various types of wetting agents. Preferred diluents for aerosol or parenteral administration are phosphate buffered saline, normal (0.9%) saline, or 5% dextrose. Compositions comprising such carriers are formulated by well-known conventional methods (see, for example, Remington's Pharmaceutical Sciences, 18th edition, A. Gennaro, ed., Mack Publishing Co., Easton, PA, 1990; and Remington, The Science and Practice of Pharmacy 20th Ed. Mack Publishing, 2000). As used herein, the singular form "a", "an", and "the" include plural references unless indicated otherwise. For example, "an" antibody includes one or more antibodies.

[0062] Where aspects or embodiments of the invention are described in terms of a Markush group or other grouping of alternatives, the present invention encompasses not only the entire group listed as a whole, but each member of the group individually and all possible subgroups of the main group. The present invention also envisages the explicit exclusion of one or more of any of the group members in the claimed invention.

[0063] Any example(s) following the term “e.g.” or “for example” is not meant to be exhaustive or limiting.

[0064] As used herein, the term “about” when used to modify a numerically defined parameter (e.g., the dose of SIRPaFc fusion protein) means that the parameter may vary by as much as 10% below or above the stated numerical value for that parameter. For example, a dose of about 5 mg means 5% ± 10%, i.e. it may vary between 4.5 mg and 5.5 mg.

[0065] SIRPaFc fusion proteins

[0066] Formulations and methods provided herein include a SIRPaFc fusion protein.

[0067] SIRPaFc fusion protein molecules are based on the extracellular region of human SIRPa. They comprise at least a region of the extracellular region of SIRPa sufficient to confer effective CD47 binding affinity and specificity. So-called “soluble” forms of SIRPa, lacking the membrane anchoring component, are described in the literature and include those referenced in WO 2010 / 070047 (Novartis), WO201 3 / 109752 (Stanford), and WO2014 / 094122 (Trillium), each incorporated by reference in its entirety.

[0068] A SIRPaFc fusion protein comprises the human SIRPa protein, in a form fused directly, or indirectly, with an antibody constant region, or Fc (fragment crystallisable). Unless otherwise stated, the term “human SIRPa” as used herein refers to a wild type, endogenous, mature form of human SIRPa. In humans, the SIRPa protein is found in two major forms. One form, the variant 1 or V1 form, has the amino acid sequence set out as NCBI RefSeq NP_542970.1 (residues 27-504 constitute the mature form). Another form, the variant 2 or V2 form, differs by 13 amino acids and has the amino acid sequence set out in GenBank as CAA71403.1 (residues 30-504 constitute the mature form). These two forms of SIRPa constitute about 80% of the forms of SIRPa present in humans, and both are embraced herein by the term “human SIRPa”. Also embraced by the term “human SIRPa” are the minor forms thereof that are endogenous to humans and have the same property of triggering signal transduction through CD47 upon binding thereto. The present invention is directed most particularly to the drug combinations that include the human SIRP variant 2 form, or V2.

[0069] In the dosing regimens and methods provided herein, useful SIRPaFc fusion proteins comprise one of the three so-called immunoglobulin (Ig) domains that lie within the extracellular region of human SIRPa. More particularly, the present SIRPaFc proteins incorporate residues 32-137 of human SIRPa (a 106-mer), which constitute and define the IgV domain of the V2 form according to current nomenclature. This SIRPa sequence, shown below, is referenced herein as SEQ ID NO: 1. EELQVIQPDKSVSVAAGESAILHCTVTSLIPVGPIQWFRGAGPARELIYNQKEGHFP RVTTVSESTKRENMDFSISISNITPADAGTYYCVKFRKGSPDTEFKSGA [SEQ ID NO: 1]

[0070] In some embodiments, SIRPaFc fusion proteins incorporate the IgV domain as defined by SEQ ID NO: 1 , and additional, flanking residues contiguous within the SIRPa sequence. This form of the IgV domain, represented by residues 31 -148 of the V2 form of human SIRPa, is a 118-mer having SEQ ID NO: 2 shown below: EEELQVIQPDKSVSVAAGESAILHCTVTSLIPVGPIQWFRGAGPARELIYNQKEGHF PRVTTVSESTKRENMDFSISISNITPADAGTYYCVKFRKGSPDTEFKSGAGTELSVR AKPS [SEQ ID NO: 2]

[0071] The present SIRPa fusion proteins can also incorporate an Fc region having effector function. Fc refers to “fragment crystallisable” and represents the constant region of an antibody comprised principally of the CH2 and CH3 domains of the heavy chain constant region and components within the hinge region. Suitable Fc components include those having effector function. An Fc component “having effector function” is an Fc component having at least some effector function, such as at least some contribution to antibody-dependent cellular cytotoxicity or some ability to fix complement. Also, the Fc will at least bind to Fc receptors. These properties can be revealed using assays established for this purpose. Functional assays include the standard chromium release assay that detects target cell lysis. By this definition, an Fc region that is wild type IgG 1 or lgG4 has effector function, whereas the Fc region of a human lgG4 mutated to eliminate effector function, such as by incorporation of an alteration series that includes Pro233, Val234, Ala235 and deletion of Gly236 (Ell), is considered not to have effector function. In some embodiments, the Fc is based on human antibodies of the lgG1 isotype. The Fc region of these antibodies will be readily identifiable to those skilled in the art. In embodiments, the Fc region includes the lower hinge-CH2-CH3 domains.

[0072] In a specific embodiment, the Fc region is based on the amino acid sequence of a human lgG1 set out as P01857 in UniProtKB / Swiss-Prot, residues 104-330, and has the amino acid sequence shown below and referenced herein as SEQ ID NO: 3: DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPA PIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPGK [SEQ ID NO: 3]

[0073] Thus, in some embodiments, the Fc region has either a wild type or consensus sequence of an lgG1 constant region. In alternative embodiments, the Fc region incorporated in the fusion protein is derived from any IgG 1 antibody having a typical effector-active constant region. The sequences of such Fc regions can correspond, for example, with the Fc regions of any of the following IgG 1 sequences (all referenced from GenBank), for example: BAG65283 (residues 242-473), BAC04226.1 (residues 247-478), BAC05014.1 (residues 240-471 ), CAC20454.1 (residues 99-320), BAC05016.1 (residues 238-469), BAC85350.1 (residues 243- 474), BAC85529.1 (residues 244-475), and BAC85429.1 (residues (238-469).

[0074] In other embodiments, the Fc region has a sequence of a wild type human lgG4 constant region. In alternative embodiments, the Fc region incorporated in the fusion protein is derived from any lgG4 antibody having a constant region with effector activity that is present but, naturally, is significantly less potent than the IgG 1 Fc region. The sequences of such Fc regions can correspond, for example, with the Fc regions of any of the following lgG4 sequences: P01861 (residues 99-327) from UniProtKB / Swiss-Prot and CAC20457.1 (residues 99-327) from GenBank. In some embodiments, the Fc region is based on the amino acid sequence of a human lgG4 set out as P01861 in UniProtKB / Swiss-Prot, residues 99-327, and has the amino acid sequence shown below and referenced herein as SEQ ID NO: 4: ESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQ FNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGL PSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQ KSLSLSLGK [SEQ ID NO: 4]

[0075] In some embodiments, the Fc region incorporates one or more alterations, usually not more than about 10, e.g., up to 1 , 2, 3, 4, 5 or 6 such alterations, including amino acid substitutions that affect certain Fc properties. In one specific embodiment, the Fc region incorporates an alteration at position 228 (Ell numbering), in which the serine at this position is substituted by a proline (S228P), thereby to stabilize the disulfide linkage within the Fc dimer. Other alterations within the Fc region can include substitutions that alter glycosylation, such as substitution of Asn297by glycine or alanine; half-life enhancing alterations such as T252L, T253S, and T256F as taught in US62777375, and many others. Particularly useful are those alterations that enhance Fc properties while remaining silent with respect to conformation, e.g., retaining Fc receptor binding. In another embodiment, the Fc region is modified to increase its biological half-life. Various approaches are possible. For example, one or more of the following mutations can be introduced; T252L, T254S, T256F, as described in U.S. Pat. No. 6,277,375.

[0076] In a specific embodiment, and in the case where the Fc component is an lgG4 Fc, the Fc incorporates at least the S228P mutation, and has the amino acid sequence set out below and referenced herein as SEQ ID NO: 5: ESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQ FNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGL PSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQ KSLSLSLGK [SEQ ID NO: 5]

[0077] The SIRPaFc fusion protein used in the regimens and methods provided herein is thus in some embodiments a SIRP fusion protein useful to inhibit the binding of human SIRPa and human CD47, thereby to inhibit or reduce transmission of the signal mediated via SIRPa-bound CD47, the fusion protein comprising a human SIRPa component and, fused therewith, an Fc component, wherein the SIRPa component comprises or consists of a single IgV domain of human SIRPa V2 and the Fc component contains a human IgG Fc domain having effector function.

[0078] In one embodiment, the fusion protein comprises a SIRPa component comprising at least of residues 32-137 of the V2 form of wild type human SIRPa, i.e. , SEQ ID NO: 1. In a preferred embodiment, the SIRPa component comprises residues 31-148 of the V2 form of human SIRPa, i.e., SEQ ID NO: 2. In one embodiment, the Fc component is the Fc component of the human IgG 1 designated P01857, and in a specific embodiment has the amino acid sequence that incorporates the lower hinge-CH2-CH3 region thereof i.e., SEQ ID NO: 3. In another embodiment, the Fc component is the Fc component of the human lgG4 designated P01861 , and in a specific embodiment has the amino acid sequence that incorporates the lower hinge-CH2-CH3 region thereof and the mutation S228P, i.e., SEQ ID NO: 5.

[0079] In some embodiments, the SIRPaFc fusion protein is provided and used in a secreted dimeric fusion form, wherein the fusion protein incorporates a SIRPa component having SEQ ID NO: 1 and preferably SEQ ID NO: 2 and, fused therewith, an Fc region having effector function and having SEQ ID NO: 3. When the SIRPa component is SEQ ID NO: 2 and the Fc region is SEQ ID NO: 3, the fusion protein comprises SEQ ID NO: 6, shown below: EEELQVIQPDKSVSVAAGESAILHCTVTSLIPVGPIQWFRGAGPARELIYNQKEGHF PRVTTVSESTKRENMDFSISISNITPADAGTYYCVKFRKGSPDTEFKSGAGTELSVR AKPSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGK [SEQ ID NO: 6]

[0080] The SIRPaFc fusion protein of SEQ ID NO: 6 is also known as TTI-621 or ontorpacept. TTI-621 I ontorpacept comprises a dimer of proteins of SEQ ID NO: 6.

[0081] In alternative embodiments, the Fc component of the fusion protein is based on an lgG4, and preferably an lgG4 that incorporates the S228P mutation. In the case where the fusion protein incorporates the preferred SIRPa IgV domain of SEQ ID NO: 2, and the lgG4 Fc region is SEQ ID NO: 5, the fusion protein comprises SEQ ID NO: 7, shown below: EEELQVIQPDKSVSVAAGESAILHCTVTSLIPVGPIQWFRGAGPARELIYNQKEGHF PRVTTVSESTKRENMDFSISISNITPADAGTYYCVKFRKGSPDTEFKSGAGTELSVR AKPSESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQED PEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVS NKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHN HYTQKSLSLSLGK [SEQ ID NO: 7]

[0082] The SIRPaFc fusion protein of SEQ ID NO: 7 is also known as TTI-622 and maplirpacept. TTI-622 I maplirpacept comprises a dimer of proteins of SEQ ID NO: 7.

[0083] In one embodiment of a dosing regimen or method provided herein, a SIRPaFc fusion protein comprises, as the SIRPa component of the fusion protein, a sequence that comprises the polypeptide of SEQ ID NO: 2. In one embodiment, the SIRPaFc fusion protein comprises the polypeptide of SEQ ID NO: 6 or SEQ ID NO: 7.

[0084] The SIRPa sequence incorporated within the SIRPaFc fusion protein can be varied, as described in the literature. This can eliminate glycosylation sites in the protein, such as at position 89 and elsewhere. Other, useful substitutions within SIRPa include one or more of the following: L4V / I, V6I / L, A21V, V27I / L, I31T / S / F, E47V / L, K53R, E54Q, H56P / R, S66T / G, K68R, V92I, F94V / L, V63I, and / or F103V.

[0085] In a SIRPaFc fusion protein, the SIRPa component and the Fc component are fused, either directly or indirectly, to provide a single chain polypeptide that may optionally be ultimately produced as a dimer in which the single chain polypeptides are coupled through inter-chain disulfide bonds formed within the Fc region. The nature of the fusing region is not critical. The fusion may be direct between the two components, with the SIRP component constituting the N-terminal end of the fusion and the Fc component constituting the C-terminal end. Alternatively, the fusion may be indirect, through a linker comprised of one or more amino acids, desirably genetically encoded amino acids, such as two, three, four, five, six, seven, eight, nine or ten amino acids, or any number of amino acids between 5 and 100 amino acids, such as between 5 and 50, 5 and 30 or 5 and 20 amino acids. A linker may comprise a peptide that is encoded by DNA constituting a restriction site, such as a BamHI, Clal, EcoRI, Hindi 11, Pstl, Sall and Xhol site and the like.

[0086] The linker amino acids typically and desirably have some flexibility to allow the Fc and the SIRP components to adopt their active conformations. Residues that allow for such flexibility typically are Gly, Asn and Ser, so that virtually any combination of these residues (and particularly Gly and Ser) within a linker is likely to provide the desired linking effect. In one example, such a linker is based on the so- called G4S sequence (Gly-Gly-Gly-Gly-Ser [SEQ ID NO: 8]) which may repeat as (G4S)n where n is 1 , 2, 3 or more, or is based on (Gly)n, (Ser)n, (Ser-Gly)n or (Gly- Ser)n and the like. In another embodiment, the linker is GTELSVRAKPS [SEQ ID NO: 9], This sequence constitutes SIRPa sequence that C-terminally flanks the IgV domain (it being understood that this flanking sequence could be considered either a linker or a different form of the IgV domain when coupled with the IgV minimal sequence described above). It is necessary only that the fusing region or linker permits the components to adopt their active conformations, and this can be achieved by any form of linker useful in the art.

[0087] SIRPaFc Fusion Protein Formulation

[0088] In one aspect, provided is a stable aqueous formulation comprising: about 10 mg / ml to about 300 mg / ml of an SIRPaFcfusion protein; a buffer; a sugar; a surfactant; a chelating agent; and wherein the formulation has a pH at about 5.5 to about 6.5. The formulation described herein have an extended shelf life, preferably of at least or more than about 3 months, 6 months, 12 months, 24 months, or 36 months (e.g., at about 5°C).

[0089] In some embodiments, the SIRPaFc fusion protein in a formulation provided herein is maplirpacept (TTI-622) or ontorpacept (TTI-621 ).

[0090] The SIRPaFc fusion protein may be present in the formulation at a concentration ranging from about 0.1 mg / ml to about 300 mg / ml, from about 15 mg / ml to 300 mg / ml, from about 20 mg / ml to about 175 mg / ml, or from about 25 mg / ml to about 150 mg / ml. For example, in some embodiments, the concentration of antibody is about 0.5 mg / ml, about 1 mg / ml, about 2 mg / ml, about 2.5 mg / ml, about 3 mg / ml, about 3.5 mg / ml, about 4 mg / ml, about 4.5 mg / ml, about 5 mg / ml, about 5.5 mg / ml, about 6 mg / ml, about 6.5 mg / ml, about 7 mg / ml, about 7.5 mg / ml, about 8 mg / ml, about 8.5 mg / ml, about 9 mg / ml, about 9.5 mg / ml, about 10 mg / ml, about 11 mg / ml, about 12 mg / ml, about 13 mg / ml, about 14 mg / ml, about 15 mg / ml, about 16 mg / ml, about 17 mg / ml, about 18 mg / ml, about 19 mg / ml, about 20 mg / ml, about 21 mg / ml, about 22 mg / ml, about 23 mg / ml, about 24 mg / ml, about 25 mg / ml, about 26 mg / ml, about 27 mg / ml, about 28 mg / ml, about 29 mg / ml, about 30 mg / ml, about 31 mg / ml, about 32 mg / ml, about 33 mg / ml, about 34 mg / ml, about 35 mg / ml, about 36 mg / ml, about 37 mg / ml, about 38 mg / ml, about 39 mg / ml, about 40 mg / ml, about 41 mg / ml, about 42 mg / ml, about 43 mg / ml, about 44 mg / ml, about 45 mg / ml, about 46 mg / ml, about 47 mg / ml, about 48 mg / ml, about 49 mg / ml, about 50 mg / ml, about 51 mg / ml, about 52 mg / ml, about 53 mg / ml, about 54 mg / ml, about 55 mg / ml, about 56 mg / ml, about 57 mg / ml, about 58 mg / ml, about 59 mg / ml, about 60 mg / ml, about 70 mg / ml, about 80 mg / ml, about 90 mg / ml, about 100 mg / ml, about 101 mg / ml, about 102 mg / ml, about 102.5 mg / ml, about 103 mg / ml, about 103.5 mg / ml, about 104 mg / ml, about 104.5 mg / ml, about 105 mg / ml, about 105.5 mg / ml, about 106 mg / ml, about 106.5 mg / ml, about 107 mg / ml, about 107.5 mg / ml, about 108 mg / ml, about 108.5 mg / ml, about 109 mg / ml, about 109.5 mg / ml, about 110 mg / ml, about 111 mg / ml, about 1 12 mg / ml, about 113 mg / ml, about 1 14 mg / ml, about 115 mg / ml, about 116 mg / ml, about 117 mg / ml, about 118 mg / ml, about 119 mg / ml, about 120 mg / ml, about 121 mg / ml, about 122 mg / ml, about 123 mg / ml, about 124 mg / ml, about 125 mg / ml, about 126 mg / ml, about 127 mg / ml, about 128 mg / ml, about 129 mg / ml, about 130 mg / ml, about 131 mg / ml, about 132 mg / ml, about 133 mg / ml, about 134 mg / ml, about 135 mg / ml, about 136 mg / ml, about 137 mg / ml, about 138 mg / ml, about 139 mg / ml, about 140 mg / ml, about 141 mg / ml, about 142 mg / ml, about 143 mg / ml, about 144 mg / ml, about 145 mg / ml, about 146 mg / ml, about 147 mg / ml, about 148 mg / ml, about 149 mg / ml, about 150 mg / ml, about 151 mg / ml, about 152 mg / ml, about 153 mg / ml, about 154 mg / ml, about 155 mg / ml, about 156 mg / ml, about 157 mg / ml, about 158 mg / ml, about 159 mg / ml, about 160 mg / ml, about 170 mg / ml, about 180 mg / ml, about 190 mg / ml, about 200 mg / ml, about 210 mg / ml, about 220 mg / ml, about 230 mg / ml, about 240 mg / ml, about 250 mg / ml, about 260 mg / ml, about 270 mg / ml, about 280 mg / ml, about 290 mg / ml, or about 300 mg / ml.

[0091] The buffer in a formulation provided herein can be, for example without limitation, acetate, succinate (e.g., disodium succinate hexahydrate), gluconate, citrate, histidine, acetic acid, phosphate, phosphoric acid, ascorbate, tartartic acid, maleic acid, glycine, lactate, lactic acid, ascorbic acid, imidazole, bicarbonate and carbonic acid, succinic acid, sodium benzoate, benzoic acid, gluconate, edetate, acetate, malate, imidazole, tris, phosphate, and mixtures thereof. In some embodiments, the buffer is histidine.

[0092] The concentration of the buffer can range from about 0.1 millimolar (mM) to about 100 mM. Optionally, the concentration of the buffer is from about 0.5 mM to about 50 mM, optionally about 1 mM to about 30 mM, optionally about 1 mM to about 25 mM. In some embodiments, the concentration of the buffer is about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 30 mM, about 35 mM, about 40 mM, about 45 mM, about 50 mM, about 55 mM, about 60 mM, about 65 mM, about 70 mM, about 75 mM, about 80 mM, about 85 mM, about 90 mM, about 95 mM, or about 100 mM. In some embodiment, the buffer is a histidine buffer in the concentration of about 20 mM.

[0093] The sugar in a formulation provided herein can be a monosaccharide, disaccharide, or polysaccharide. In some embodiments, the sugar is sucrose, trehalose, fructose, glucose, mannose, or lactose.

[0094] The concentration of the sugar in the formulation provided herein ranges from about 1 mg / ml to about 300 mg / ml, from about 1 mg / ml to about 200 mg / ml, or from about 1 mg / ml to about 120 mg / ml. Preferably the concentration of the sugar in the formulation is about 50 mg / ml to about 120 mg / ml, from about 60 mg / ml to about 1 10 mg / ml, or from about 80 mg / ml to about 90 mg / ml). For example, the concentration of the sugar in the formulation is about 0.5 mg / ml, about 1 mg / ml, about 2 mg / ml, about 2.5 mg / ml, about 3 mg / ml, about 3.5 mg / ml, about 4 mg / ml, about 4.5 mg / ml, about 5 mg / ml, about 5.5 mg / ml, about 6 mg / ml, about 6.5 mg / ml, about 7 mg / ml, about 7.5 mg / ml, about 8 mg / ml, about 8.5 mg / ml, about 9 mg / ml, about 9.5 mg / ml, about 10 mg / ml, about 11 mg / ml, about 12 mg / ml, about 13 mg / ml, about 14 mg / ml, about 15 mg / ml, about 16 mg / ml, about 17 mg / ml, about 18 mg / ml, about 19 mg / ml, about 20 mg / ml, about 21 mg / ml, about 22 mg / ml, about 23 mg / ml, about 24 mg / ml, about 25 mg / ml, about 26 mg / ml, about 27 mg / ml, about 28 mg / ml, about 29 mg / ml, about 30 mg / ml, about 31 mg / ml, about 32 mg / ml, about 33 mg / ml, about 34 mg / ml, about 35 mg / ml, about 36 mg / ml, about 37 mg / ml, about 38 mg / ml, about 39 mg / ml, about 40 mg / ml, about 41 mg / ml, about 42 mg / ml, about 43 mg / ml, about 44 mg / ml, about 45 mg / ml, about 46 mg / ml, about 47 mg / ml, about 48 mg / ml, about 49 mg / ml, about 50 mg / ml, about 51 mg / ml, about 52 mg / ml, about 53 mg / ml, about 54 mg / ml, about 55 mg / ml, about 56 mg / ml, about 57 mg / ml, about 58 mg / ml, about 59 mg / ml, about 60 mg / ml, about 65 mg / ml, about 70 mg / ml, about 75 mg / ml, about 80 mg / ml, about 81 mg / ml, about 82 mg / ml, about 83 mg / ml, about 84 mg / ml, about 85 mg / ml, about 86 mg / ml, about 87 mg / ml, about 88 mg / ml, about 89 mg / ml, about 90 mg / ml, about 91 mg / ml, about 92 mg / ml, about 93 mg / ml, about 94 mg / ml, about 95 mg / ml, about 96 mg / ml, about 97 mg / ml, about 98 mg / ml, about 99 mg / ml, about 100 mg / ml, about 101 mg / ml, about 102 mg / ml, about 103 mg / ml, about 104 mg / ml, about 105 mg / ml, about 106 mg / ml, about 107 mg / ml, about 108 mg / ml, about 109 mg / ml, about 110 mg / ml, about 1 11 mg / ml, about 112 mg / ml, about 1 13 mg / ml, about 114 mg / ml, about 115 mg / ml, about 116 mg / ml, about 117 mg / ml, about 118 mg / ml, about 119 mg / ml, about 120 mg / ml, about 121 mg / ml, about 122 mg / ml, about 123 mg / ml, about 124 mg / ml, about 125 mg / ml, about 126 mg / ml, about 127 mg / ml, about 128 mg / ml, about 129 mg / ml, about 130 mg / ml, about 131 mg / ml, about 132 mg / ml, about 133 mg / ml, about 134 mg / ml, about 135 mg / ml, about 136 mg / ml, about 137 mg / ml, about 138 mg / ml, about 139 mg / ml, about 140 mg / ml, about 141 mg / ml, about 142 mg / ml, about 143 mg / ml, about 144 mg / ml, about 145 mg / ml, about 146 mg / ml, about 147 mg / ml, about 148 mg / ml, about 149 mg / ml, or about 150 mg / ml.

[0095] In some embodiments, the sugar is sucrose at a concentration of from about 1 mg / ml to about 300 mg / ml, from about 1 mg / ml to about 200 mg / ml, or from about 1 mg / ml to about 120 mg / ml. In some embodiments, the concentration of the sucrose in the formulation is about 50 mg / ml to about 120 mg / ml, from about 60 mg / ml to about 110 mg / ml, or from about 80 mg / ml to about 90 mg / ml. In some embodiments, the concentration of sucrose in the formulation is about 60 mg / ml.

[0096] Surfactants, as used in the formulations provided herein, can alter the surface tension of a liquid SIRPaFc fusion protein formulation. In certain embodiments, the surfactant reduces the surface tension of a liquid SIRPaFc formulation. In still other embodiments, the surfactant can contribute to an improvement in stability of any of the SIRPaFc in the formulation. The surfactant can also reduce aggregation of the formulated SIRPaFc (e.g., during shipping and storage) and / or minimize the formation of particulates in the formulation and / or reduces adsorption (e.g., adsorption to a container). For example, the surfactant can also improve stability of the SIRPaFc during and after a freeze / thaw cycle. The surfactant can be, for example without limitation, a polysorbate, poloxamer, triton, sodium dodecyl sulfate, sodium laurel sulfate, sodium octyl glycoside, lauryl-sulfobetaine, myristyl-sulfobetaine, linoleyl- sulfobetaine, stearyl-sulfobetaine, lauryl-sarcosine, myristyl-sarcosine, linoleyl- sarcosine, stearyl-sarcosine, linoleyl-betaine, myristyl-betaine, cetyl-betaine, lauroamidopropyl-betaine, cocam idopropyl-betaine, linoleamidopropyl-betaine, myristamidopropyl-betaine, palmidopropyl-betaine, isostearamidopropyl-betaine, myristamidopropyl-dimethylamine, palmidopropyl-dimethylamine, isostearamidopropyl-dimethylamine, sodium methyl cocoyl-taurate, disodium methyl oleyl- taurate, dihydroxypropyl PEG 5 linoleammonium chloride, polyethylene glycol, polypropylene glycol, and mixtures thereof. The surfactant can be, for example without limitation, polysorbate 20, polysorbate 21 , polysorbate 40, polysorbate 60, polysorbate 61 , polysorbate 65, polysorbate 80, polysorbate 81 , polysorbate 85, PEG3350 and mixtures thereof.

[0097] The concentration of the surfactant generally ranges from about 0.01 mg / ml to about 10 mg / ml, from about 0.01 mg / ml to about 5.0 mg / ml, from about 0.01 mg / ml to about 2.0 mg / ml, from about 0.01 mg / ml to about 1 .5 mg / ml, from about 0.01 mg / ml to about 1 .0 mg / ml, from about 0.01 mg / ml to about 0.5 mg / ml, from about 0.01 mg / ml to about 0.4 mg / ml, from about 0.01 mg / ml to about 0.3 mg / ml, from about 0.01 mg / ml to about 0.2 mg / ml, from about 0.01 mg / ml to about 0.15 mg / ml, from about 0.01 mg / ml to about 0.1 mg / ml, from about 0.01 mg / ml to about 0.05 mg / ml, from about 0.1 mg / ml to about 1 mg / ml, from about 0.1 mg / ml to about 0.5 mg / ml, or from about 0.1 mg / ml to about 0.3 mg / ml. Further preferably the concentration of the surfactant is about 0.05 mg / ml, about 0.06 mg / ml, about 0.07 mg / ml, about 0.08 mg / ml, about 0.09 mg / ml, about 0.1 mg / ml, about 0.15 mg / ml, about 0.2 mg / ml, about 0.3 mg / ml, about 0.4 mg / ml, about 0.5 mg / ml, about 0.6 mg / ml, about 0.7 mg / ml, about 0.8 mg / ml, about 0.9 mg / ml, or about 1 mg / ml.

[0098] In some embodiments, the polysorbate is polysorbate 80 at a concentration ranging from about 0.1 mg / ml to about 0.3 mg / ml, for example, at 0.2 mg / ml.

[0099] Chelating agents, as used in formulations provided herein, lower the formation of reduced oxygen species, reduce acidic species (e.g., deamidation) formation, reduce antibody aggregation, and / or reduce antibody fragmentation, and / or reduce antibody oxidation in the formulation of the present invention. For example, the chelating agent can be a multidentate ligand that forms at least one bond (e.g., covalent, ionic, or otherwise) to a metal ion and acts as a stabilizer to complex with species, which might otherwise promote instability.

[0100] In some embodiments, the chelating agent can be selected from the group consisting of aminopolycarboxylic acids, hydroxyaminocarboxylic acids, N-substituted glycines, 2- (2-amino-2-oxocthyl) aminoethane sulfonic acid (BES), deferoxamine (DEF), citric acid, niacinamide, and desoxycholates and mixtures thereof. In some embodiments, the chelating agent is selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), diethylenetriamine pentaacetic acid 5 (DTPA), nitrilotriacetic acid (NTA), N-2-acetamido-2-iminodiacetic acid (ADA), bis(aminoethyl)glycolether, N,N,N',N'-tetraacetic acid (EGTA), trans- diaminocyclohexane tetraacetic acid (DCTA), glutamic acid, and aspartic acid, N- hydroxyethyliminodiacetic acid (HIMDA), N,N-bis-hydroxyethylglycine (bicine) and N- (trishydroxymethylmethyl) 10 glycine (tricine), glycylglycine, sodium desoxycholate, ethylenediamine, propylenediamine, diethylenetriamine, triethylenetetraamine (then), disodium edetate dihydrate (or disodium EDTA dihydrate or EDTA disodium salt), calcium EDTA oxalic acid, malate, citric acid, citric acid monohydrate, and trisodium citrate-dihydrate, 8-hydroxyquinolate, amino acids, histidine, cysteine, methionine, peptides, polypeptides, and proteins and mixtures thereof. In some embodiments, the chelating agent is selected from the group consisting of salts of EDTA including dipotassium edetate, disodium edetate, edetate calcium disodium, sodium edetate, trisodium edetate, and potassium edetate; and a suitable salt of deferoxamine (DEF) is deferoxamine mesylate (DFM), or mixtures thereof. Chelating agents used in the invention can be present, where possible, as the free acid or free base form or salt form of the compound, also as an anhydrous, solvated or hydrated form of the compound or corresponding salt.

[0101] In some embodiments, the chelating agent is EDTA (edetate).

[0102] In some embodiments, the concentration of the chelating agent ranges from about 0.01 mg / ml to about 50 mg / ml, from about 0.1 mg / ml to about 10.0 mg / ml, from about 5 mg / ml to about 15.0 mg / ml, from about 0.01 mg / ml to about 1.0 mg / ml, from about 0.02 mg / ml to about 0.5 mg / ml, from about 0.025 mg / ml to about 0.075 mg / ml. Optionally, the concentration of the chelating agent ranges from about 0.01 mM to about 2.0 mM, from about 0.01 mM to about 1 .5 mM, from about 0.01 mM to about 0.5 mM, from about 0.01 mM to about 0.4 mM, from about 0.01 mM to about 0.3 mM, from about 0.01 mM to about 0.2 mM, from about 0.01 mM to about 0.15 mM, from about 0.01 mM to about 0.1 mM, from about 0.01 mM to about 0.09 mM, from about 0.01 mM to about 0.08 mM, from about 0.01 mM to about 0.07 mM, from about 0.01 mM to about 0.06 mM, from about 0.01 mM to about 0.05 mM, from about 0.01 mM to about 0.04 mM, from about 0.01 mM to about 0.03 mM, from about 0.01 mM to about 0.02 mM, from about 0.02, or from about 0.05 mM to about 0.01 mM. In some embodiments, the concentration of the chelating agent can be about 0.01 mg / ml, about 0.02 mg / ml, about 0.025 mg / ml, about 0.03 mg / ml, about 0.04 mg / ml, about 0.05 mg / ml, about 0.06 mg / ml, about 0.07 mg / ml, about 0.075 mg / ml, about 0.08 mg / ml, about 0.09 mg / ml, about 0.10 mg / ml, or about 0.20 mg / ml. In some embodiments, the concentration of chelating agent is about 0.025 mg / ml, about 0.03 mg / ml, about 0.035 mg / ml, about 0.04 mg / ml, about 0.045 mg / ml, about 0.05 mg / ml, about 0.055 mg / ml, about 0.06 mg / ml, about 0.065 mg / ml, about 0.07 mg / ml, or about 0.075 mg / ml. In some embodiments, the concentration of the chelating agent is about 0.05 mg / ml.

[0103] According to some embodiments of the present invention, the pH can be in the range of about pH 5.0 to about 7.0, optionally between about pH 5.5 to 6.5 or about 5.8 to 6.2, or between pH 5.9 to 6.1. For example, the SIRPaFc fusion protein in the formulation of the present invention at the pH range of 6.0 had less formation of high molecular mass species compared to at pH 5.0 or pH 7.0. Accordingly, in some embodiments, the pH for the formulation of the present invention can be in the range selected from between any one of about pH 5.5, 5.6 5.7, 5.8, or 5.9 and any one of about pH 6.1 , 6.2, 6.3, 6.4, or 6.5. In some embodiments the pH can be selected from pH values of any of about pH 5.0, 5.1 , 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1 , 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1 , 7.2, 7.3, 7.4 or 7.5. In some embodiments, the pH is pH 6.0 + / - 0.5, pH 6.0 + / - 0.3, 6.0 + / - 0.2, or pH 6.0 + / - 0.1.

[0104] In some embodiments the formulation can comprise a preservative. In some embodiments, the preservative agent is selected from phenol, m-cresol, benzyl alcohol, benzalkonium chloride, benzalthonium chloride, phenoxyethanol and methyl paraben.

[0105] In some embodiments, the formulation does not contain a preservative.

[0106] In some embodiments, a formulation as described herein has a shelf life of at least or more than about 6 months, 12 months, 18 months, 24 months, 30 months, 36 months, 42 months, or 48 months (e.g., at 5°C, 25°C, or 40°C). For example, in some embodiments, a formulation provided herein has a shelf life of at least about 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 24 months, 25 months, 26 months, 27 months, 28 months, 29 months, 30 months, 31 months, 32 months, 33 months, 34 months, 35 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, or 60 months (e.g., at 5°C, 25°C, or 40°C).

[0107] In some embodiments, the formulation as described herein has less than about 5% HMMS at 40°C for up to 1 month, 2 months, 3 months, 4 months, 5 months, or 6 months, or up to 4 weeks, 8 weeks, 12 weeks, 16 weeks, 20 weeks, or 24 weeks (e.g., as measured by size exclusion HPLC). In some embodiments, the formulation as described herein has less than about 5% HMMS at 2-8°C for up to 36 months (e.g., as measured by size exclusion HPLC).

[0108] Unless stated otherwise, the concentrations listed herein are those concentrations at ambient conditions, i.e. , at 25°C and atmospheric pressure.

[0109] Methods of Using the SIRPaFc Fusion Protein Formulation

[0110] The formulations of the present invention are useful in various applications including, but are not limited to, therapeutic treatment methods.

[0111] In one aspect, the invention provides a method for treating diseases and pathological conditions. In particular, the invention provides an effective approach for treating cancers. Accordingly, in some embodiments, provided is a method of treating or inhibiting cancer in a subject in need thereof comprising administering to the subject a therapeutically effective amount of the formulation as described herein. In some embodiments, provided is the formulation of the present invention for use in the treatment of cancer. In some embodiments, provided is a use of the formulation of the present invention for the manufacture of a medicament for treatment of cancer.

[0112] In some embodiments, a SIRPaFc fusion protein formulation as provided herein can be used with an additional anticancer agent.

[0113] Dosing regimens and methods provided herein may be is useful to treat a variety of cancer cells. These include particularly CD47+cancer cells, including liquid (hematological) and solid tumours. Solid tumours can be treated with the dosing regimens and methods provided herein, to reduce the size, number or growth rate thereof and to control growth of cancer stem cells. Such solid tumours include CD47+tumours in bladder, brain, breast, lung, colon, ovary, prostate, liver and other tissues as well. In one embodiment, dosing regimens and methods provided herein can used to inhibit the growth or proliferation of hematological cancers. As used herein, “hematological cancer” refers to a cancer of the blood, and includes leukemia, lymphoma and myeloma among others. “Leukemia” refers to a cancer of the blood, in which too many white blood cells that are ineffective in fighting infection are made, thus crowding out the other parts that make up the blood, such as platelets and red blood cells. It is understood that cases of leukemia are classified as acute or chronic. Certain forms of leukemia may be, by way of example, acute lymphocytic leukemia (ALL); acute myeloid leukemia (AML); chronic lymphocytic leukemia (CLL); chronic myelogenous leukemia (CML); myeloproliferative disorder / neoplasm (MPDS); and myelodysplastic syndrome. “Lymphoma” may refer to a Hodgkin’s lymphoma, both indolent and aggressive non-Hodgkin’s lymphoma, Burkitt's lymphoma, and follicular lymphoma (small cell and large cell), among others. Myeloma may refer to multiple myeloma (MM), giant cell myeloma, heavychain myeloma, and light chain or Bence-Jones myeloma. In particular embodiments, dosing regimens and methods provided herein are useful to treat T cell lymphomas that are a very heterogeneous group of lymphoid malignancies divided into cutaneous and peripheral TCL, which themselves are divided into nodal or extranodal types. CTCL derive from skin-homing T cells and consist of mycosis fungoides, Sezary syndrome, primary cutaneous T cell lymphoproliferative disorders, and anaplastic large cell lymphoma. The common features of TCL are aggressive course and poor response to therapy, with the exception of ALK and ALCL.

[0114] In some other embodiments, the hematological cancer treated with dosing regimens and methods is a CD47+leukemia, preferably selected from acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, and myelodysplastic syndrome, preferably, human acute myeloid leukemia.

[0115] In other embodiments, the hematological cancer treated with a dosing regimen or method provided herein is a CD47+lymphoma or myeloma selected from Hodgkin’s lymphoma, both indolent and aggressive non-Hodgkin’s lymphoma, Burkitt's lymphoma, follicular lymphoma (small cell and large cell), multiple myeloma (MM), giant cell myeloma, heavy-chain myeloma, and light chain or Bence-Jones myeloma as well as leimyosarcoma.

[0116] In some embodiments, a cancer treated with a dosing regimen or method provided herein is relapsed and / or refractory (R / R). In some embodiments, a subject treated with a dosing regimen or method provided herein has been previously treated with 1 -3 lines of therapy for the cancer.

[0117] In some embodiments, the formulation of the present invention can be administered directly into the blood stream, into muscle, into tissue, into fat, or into an internal organ of a subject. Suitable means for parenteral administration include intravenous, intraocular, intravitreal, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, intrasternal, intracranial, intramuscular, intra-ossial, intradermal and subcutaneous. Suitable devices for parenteral administration include needle (including microneedle, microprojections, soluble needles and other micropore formation techniques) injectors, needle-free injectors and infusion techniques. In some embodiments, the formulation of the present invention is administered to the subject intravenously or subcutaneously.

[0118] In some embodiments, the administration pattern of the formulation of the present invention comprises administration of a dose of the formulation once every week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every seven weeks, once every eight weeks, once every nine weeks, once every ten weeks, once every fifteen weeks, once every twenty weeks, once every twenty five weeks, or once every twenty six weeks. In some embodiments, the formulation of the present invention is administered once every month, once every two months, once every three months, once every four months, once every five months, or once every six months.

[0119] Dosage regimens may depend on the pattern of pharmacokinetic decay that the practitioner wishes to achieve. For example, in some embodiments, dosing from one-four times a week is contemplated. Even less frequent dosing may be used. In some embodiments, the dose is administered once every 1 week, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, every 9 weeks, every 10 weeks, every 15 weeks, every 20 weeks, every 25 weeks, or longer. In some embodiments, the dose is administered once every 1 month, every 2 months, every 3 months, every 4 months, every 5 months, every 6 months, or longer. The following examples are offered for illustrative purposes only, and are not intended to limit the scope of the present invention in any way. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and fall within the scope of the appended claims.

[0120] Examples

[0121] Example 1 : Screening Study of Impact of pH and Buffers on the Physical and Chemical Stability of SIRPa-Fc fusion protein

[0122] To evaluate possible SIRPaFc fusion protein formulations, a screening study to evaluate the impact of different buffer systems and pH on the physical (aggregation, denaturation) and chemical stability of maplirpacept at 150 mg / mL was performed.

[0123] To perform the study, maplirpacept at 150 mg / mL was prepared in the different buffer and pH conditions shown in Table 1. All conditions also contain 150 mM NaCI.

[0124] Table 1 :

[0125] Maplirpacept in Conditions 1-6 were placed into 25 °C and 40 °C temperature- controlled chambers. Samples from each Condition were pulled at different storage times [0 hours (TO), 1 week (T1W), and 2 weeks (T2W)] to test the sample’s quality attributes. pH

[0126] The target pH for Conditions 1 , 2, 3, 4, 5, and 6 were 4.0, 5.0, 6.0, 7.0, 8.0, and 7.4 respectively. TO samples were tested for pH. Samples from Conditions 1 and 2 were pH 4.4. There were no significant changes between the target pH and the TO pH measurement for Conditions 3-6.

[0127] SEC

[0128] The presence of monomers and high molecular mass species (HMMS) was monitored using Size Exclusion Chromatography (SEC). “Monomers” in this Example and Example 2 refers to soluble (i.e. non-aggregated) maplirpacept molecules. After two weeks at 25 °C, there was a significant change (approximately 4-6% decrease) in % monomer for pH 4.0 and 5.0 samples and no significant change (< 1 % decrease) for pH 6.0, 7.0, 8.0, and 7.4 target samples (FIG. 1A). After two weeks at 40 °C (accelerated condition), all the formulations had significantly decreased monomer content (FIG. 1 B). After one week at 40 °C, samples from target pH 4.0 and 5.0 formed a gel and therefore were not tested. After two weeks at 40 °C, samples from target pH 6.0, 7.0, 8.0 and 7.4 presented a decrease of 2.6, 4.5, 7.7, and 6.2 % in monomer content, respectively (FIG. 1 B). iCE

[0129] Imaged Capillary Isoelectric Focusing (iCE) quantifies the differently charged species in a sample. At Time 0 (TO), the % acidic species was ~39% in all six tested formulations. At 2 weeks of storage at 25 °C, a decrease in the % of acidic species for the formulations of pH 4.0 (-1.8), 5.0 (-3.0), 6.0 (-1.1) and 7.0 (0.4) and an increase in the % of acidic species for the formulations of pH 8.0 (+1.6) and 7.4 (+1.5) was observed (FIG. 2A). At two weeks of storage at 40 °C, an increase in the % of acidic species was observed and more pronounced in samples with higher pH (FIG. 2B).

[0130] Conclusions

[0131] The results from these studies indicate that the formulation at pH 6.0 showed the fewest changes as measured by SEC and iCE assays when incubated at 25 °C and 40 °C for two weeks. Example 2: SIRPaFc Fusion Protein Formulation Nomination Study

[0132] To further evaluate possible SIRPaFc fusion protein formulations, an optimization study to evaluate the impact of sucrose or NaCI with 150 or 200 mg / mL maplirpacept in buffer containing 20 mM histidine, pH 6.0 on the stability during various environmental stressors was performed.

[0133] To perform the study, maplirpacept was prepared in the different formulation conditions shown in Table 2. All conditions also contain 0.05 g / L EDTA, 0.2 g / L polysorbate 80 (PS80), and 20 mM histidine, pH 6.0.

[0134] Table 2:

[0135] The different maplirpacept formulations were exposed to three types of environmental stressors: i) 24 hours of agitation; ii) five freeze and thaw cycles; and iii) thermal stability at four different temperatures.

[0136] For the agitation (Ag) test, samples were pulled from the -70 °C freezer, set out to room temperature and allowed to equilibrate. The samples were then exposed to agitation at 300 rotations per minute (RPM) for 24 hours. After the agitation period, samples were frozen at -70 °C until testing.

[0137] For the freeze-thaw test, samples were pulled from the -70 °C freezer, set out to room temperature and allowed to equilibrate. Samples were then placed at -20 °C overnight, then thawed the following morning at 25 °C. Once thawed and equilibrated to room temperature, the samples were returned to the -20 °C freezer. This process was repeated for a total of five freeze I thaw cycles. Once the freeze I thaw cycles were completed, the samples were placed in a -70 °C freezer until testing.

[0138] For the thermal stability test, samples for each buffer condition were labeled and separated out into Time zero (starting point)(T0), two weeks (T2W), four weeks (T4W), eight weeks (T8W), and twelve weeks (T12W) sets for each of the following temperature conditions: -20 °C, 5 °C, 25 °C, and 40 °C. All samples were placed in the designated stability chambers.

[0139] Results

[0140] Appearance

[0141] Visual appearance including color, clarity, and presence of visible particles of all samples was observed in a light box where the lux was measured using a traceable light meter. All samples were consistent throughout the five freeze I thaw cycles, 24 hours of agitation at 300 rpm, 12 weeks at -20 °C, 5 °C, and 25 °C, and up to 4 weeks at 40 °C. The placebo condition was slightly less intense in color as compared to samples that contained maplirpacept. No particulate formation was observed over the course of the study in samples containing maplirpacept.

[0142] Bioassav ELISA

[0143] Potency was determined by bioassay ELISA. Relative Potency (%) remained consistent with expected values throughout the study for all conditions.

[0144] CGE non-reduced

[0145] Analysis by capillary gel electrophoresis (CGE) under non-reducing conditions was performed to quantify the percentage of species with intact intrachain and interchain IgG structure compared to fragmented and other species. All peak shapes were representative of the reference material and there was no significant increase in fragmented species in any sample tested.

[0146] CGE reduced

[0147] Analysis by CGE under reducing conditions was performed to quantify the percentage of intact heavy and light chains compared to fragmented and other species. All peak shapes were representative of the reference material and no significant increase in fragmented or other species was observed in any -20 °C or 5 °C samples, and only a mild increase was shown in the stressed conditions (25 °C and 40 °C). iCE Charge heterogeneity was quantified using iCE. No significant increase in acidic species was observed in any of the -20 °C, 5 °C, or 25 °C samples. Small increases in acid species were observed in 40 °C samples.

[0148] SEC

[0149] Size exclusion chromatography (SEC) was quantified using high performance liquid chromatography (HPLC). At -20 °C both M03 and M05 exhibited no increase in % HMMS through 12 weeks. Both formulations had similar, minor decrease in % monomer between 8 weeks and 12 weeks (M03: monomer decreased by 2%; M05: monomer decreased by 1 .9%). At 5°C and 25°C M03 and M05 were similarly stable up to 8 weeks. However, between 8 weeks and 12 weeks M05 degraded at a slightly faster rate. At 12 weeks, 5°C M05 % monomer decreased by 2.7 %, where M03 % monomer decreased by 2.2%. At 12 weeks, 25 °C M05 % monomer decreased by 3.0%, where M03 % monomer decreased by 2.7%. M04 followed a similar trend: stable up to 8 weeks, then minor degradation showing at 12 weeks, with a slightly higher degradation rate than M03. M04 overall showed to be less stable than M03 or M05 at all time points.

[0150] Conclusions

[0151] Overall, these assays show that maplirpacept is stable at 150 mg / mL and 200 mg / mL over 12 weeks at -20 °C, 5 °C, and 25 °C, for five freeze I thaw cycles, and for 24 hours of agitation at 300 rpm. Throughout testing across all time points and temperatures, formulation conditions M03 (sucrose) and M05 (NaCI) performed comparably. SEC data at 5 °C shows that M03 had less HMMS than M05 at 12 weeks.

[0152] Although the disclosed teachings have been described with reference to various applications, methods, kits, and compositions, it will be appreciated that various changes and modifications can be made without departing from the teachings herein and the claimed invention below. The foregoing examples are provided to better illustrate the disclosed teachings and are not intended to limit the scope of the teachings presented herein. While the present teachings have been described in terms of these exemplary embodiments, the skilled artisan will readily understand that numerous variations and modifications of these exemplary embodiments are possible without undue experimentation. All such variations and modifications are within the scope of the current teachings.

[0153] All references cited herein, including patents, patent applications, papers, textbooks, and the like, and the references cited therein, to the extent that they are not already, are hereby incorporated by reference in their entirety. In the event that one or more of the incorporated literature and similar materials differs from or contradicts this application, including but not limited to defined terms, term usage, described techniques, or the like, this application controls.

[0154] The foregoing description and Examples detail certain specific embodiments of the invention and describes the best mode contemplated by the inventors. It will be appreciated, however, that no matter how detailed the foregoing may appear in text, the invention may be practiced in many ways and the invention should be construed in accordance with the appended claims and any equivalents thereof.

Claims

ClaimsIt is claimed:

1. An aqueous formulation comprising: 10 mg / mL to 300 mg / mL of a SIRPalpha-Fc fusion protein (SIRPaFc), a buffer, a sugar, a surfactant, and a chelating agent, wherein the formulation has a pH of 5.5 to 6.5.

2. The aqueous formulation of claim 1 , wherein the buffer is a histidine buffer.

3. The aqueous formulation of any one of claims 1-2, wherein the concentration of the buffer is 5-50 mM.

4. The aqueous formulation of any one of claims 1-3, wherein the sugar is sucrose.

5. The aqueous formulation of any one of claims 1-4, wherein the concentration of the sugar is about 1 mg / ml to about 300 mg / ml.

6. The aqueous formulation of any one of claims 1-5, wherein the surfactant is a polysorbate.

7. The aqueous formulation of claim 6, wherein the polysorbate is polysorbate 80 (PS80).

8. The aqueous formulation of claim 7, wherein the concentration of the surfactant is 0.01 mg / ml to 10 mg / ml.

9. The aqueous formulation of any one of claims 1 to 8, wherein the chelating agent is ethylenediaminetetracetic acid (EDTA).

10. The aqueous formulation of any one of claims 1 to 9, wherein the concentration of the chelating agent is about 0.01 mg / ml to about 1.0 mg / ml.11 . The aqueous formulation of any one of claims 1 -10, wherein the SIRPa portion of the SIRPaFc comprises the amino acid sequence shown in SEQ ID NO: 2.

12. The aqueous formulation of any one of claims 1 -11 , wherein the SIRPaFc comprises the amino acid sequence as shown in SEQ ID NO: 6 or SEQ ID NO: 7.

13. The aqueous formulation of any one of claims 1 -12, wherein the formulation has a shelf life of at least 12 weeks.

14. An aqueous formulation comprising:10 mg / mL to 300 mg / mL of a SIRPaFc;5-50 mM of a histidine buffer;1 mg / ml to 300 mg / ml of sucrose;0.01 mg / ml to 10 mg / ml of PS80;0.01 mg / ml to 1 mg / ml of EDTA; and wherein the formulation has a pH of 5.5 to 6.5.

15. The aqueous formulation of claim 14, wherein the SIRPaFc comprises the amino acid sequence as shown in SEQ ID NO: 6 or SEQ ID NO: 7.

16. The aqueous formulation of any one of claims 1 -15, wherein the SIRPaFc is at a concentration of 50 mg / ml, 75 mg / ml, 100 mg / ml, 125 mg / ml, 150 mg / ml, 175 mg / ml, or 200 mg / ml.

17. An aqueous formulation comprising:150 mg / ml of a SIRPaFc;20 mM histidine;60 mg / ml sucrose;0.2 mg / ml PS80;0.05 mg / ml EDTA; wherein the SIRPaFc comprises the amino acid sequence as shown in SEQ ID NO:7 and wherein the formulation has a pH of 6.

18. The aqueous formulation of any one of claims 1-17, wherein the formulation has a shelf life of at least 12 weeks.

19. A method for treating or inhibiting cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the aqueous formulation of any one of claims 1 to 18.

20. The method of claim 19, wherein the cancer is a hematological cancer.

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