Stable formulations of anti-ILT4 antibodies or antigen-binding fragments thereof in combination with anti-PD-1 antibodies and methods of use thereof

Abstract

The present invention relates to stable co-formulations of an antibody or antigen-binding fragment thereof that binds to human immunoglobulin-like transcript 4 (ILT4) and PD-1. Also provided are methods of treating various cancers using the formulations disclosed herein.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 63 / 336,839, filed April 29, 2022, the disclosure of which is incorporated herein in its entirety.

[0002] REFERENCE TO ELECTRONICALLY SUBMITTED SEQUENCE LISTING This application contains a Sequence Listing that has been submitted electronically in XML format and is incorporated by reference in its entirety. The XML file was created on October 17, 2022, has the filename 25400-WO-PCT_SL.xml, and is 99,833 bytes in size.

[0003] The present invention relates generally to pharmaceutical formulations of antibodies or antigen-binding fragments thereof that bind to human interleukin-like transcript 4 (ILT4) and their use in the treatment of various diseases (eg, cancer). [Background technology]

[0004] A common strategy used by tumor cells to evade innate and adaptive immune responses is associated with aberrant expression of human leukocyte antigen (HLA)-G (Curigliano et al. Clin Cancer Res. 2013; 19(20): 5564-5571 and Gonzalez et al. Crit Rev Clin Lab Sci. 2012; 49(3): 63-84). HLA-G can directly inhibit immune cell function through receptor binding and / or through impaired trogocytosis and chemotaxis (Morandi et al. Cytokine Growth Factor Review. 2014, 25: 327-335 and Lin et al. Mol Med. 2015, 21: 782-791). For example, high expression of HLA-G in multiple types of tumors, including colorectal, pancreatic, endometrial, lung, breast, ovarian and gastric cancer, is associated with advanced disease stage, tumor invasiveness, metastatic potential and poor prognosis (Lin et al. Mol Med. 2015, 21: 782-791; and Loumange et al. Int J Cancer. 2014, 135, 222: 581-597). Antibody-mediated blockade of HLA-G function in transgenic mouse models has been shown to inhibit tumor development and block myeloid-derived suppressor cell (MDSC) expansion (Loumange et al. Int J Cancer. 2014, 135, 222: 581-597; Lin et al. Hum Immunol. 2013, 74: 439-446; and Agaugue et al. Blood. 2011, 117: 7021-7031). Binding of HLA-G to ILT4 can directly inhibit the function of monocytes, dendritic cells, and neutrophils, thus potentially compromising the innate immune antitumor response.Interaction of HLA-G with monocytes through ILT4 inhibits maturation of human monocyte-derived antigen-presenting cells (APCs), resulting in reduced expression of MHC class II antigens and costimulatory molecules through activation of Stat3 (Colonna et al. J Immunol. 1998, 160: 3096-3100; Allan et al. J Exp Med. 1999, 189(7): 1149-1155; and Liang et al. Proc Natl Sci USA. 2008, 105(24): 8357). Using human monocyte-derived dendritic cells (DCs) and ILT4 transgenic mice, it was shown that HLA-G inhibits the maturation / activation of myeloid DCs and induces the generation of tolerogenic APCs, and the induction of tolerogenic DCs by HLA-G was mediated through the disruption of the MHC class II presentation pathway (Ristich et al. Eur J Immunol. 2005, 35: 1133-1142).

[0005] PD-1 is recognized as a key molecule in immune regulation and the maintenance of peripheral immune tolerance. PD-1 is moderately expressed on naive T cells, B cells, and NKT cells, and is upregulated by T / B cell receptor signaling in lymphocytes, monocytes, and myeloid cells. Two known ligands for PD-1, PD-L1 (B7-H1) and PD-L2 (B7-DC), are expressed in human cancers occurring in a variety of tissues. In large sample sets, including ovarian, renal, colorectal, pancreatic, hepatic, and melanoma, expression of PD-L1 has been shown to correlate with poor prognosis and reduced overall survival, regardless of subsequent treatment. Similarly, expression of PD-1 on tumor-infiltrating lymphocytes has been found to characterize dysfunctional T cells in breast cancer and melanoma, and to correlate with poor prognosis in renal cancer. Thus, it has been proposed that tumor cells that express PD-L1 interact with T cells that express PD-1 to attenuate T cell activation and evade immune surveillance, thereby contributing to a reduced immune response against the tumor.

[0006] Several monoclonal antibodies that block the interaction of PD-1 with one or both of its ligands, PD-L1 and PD-L2, have been approved by the FDA for the treatment of cancer. It has been proposed that the efficacy of such antibodies may be enhanced when administered in combination with other approved or experimental cancer treatments (e.g., radiation, surgery, chemotherapy agents, targeted therapies, agents that block other unregulated signaling pathways in tumors, and other immune enhancing agents). [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Curigliano et al. Clin Cancer Res. 2013; 19(20): 5564-5571 [Non-Patent Document 2] Gonzalez et al. Crit Rev Clin Lab Sci. 2012; 49(3): 63-84 [Non-Patent Document 3] Morandi et al. Cytokine Growth Factor Review. 2014, 25: 327-335 [Non-Patent Document 4] Lin et al. Mol Med. 2015, 21: 782-791 [Non-Patent Document 5] Loumange et al. Int J Cancer. 2014, 135, 222: 581-597 [Non-Patent Document 6] Lin et al. Hum Immunol. 2013, 74: 439-446 [Non-Patent Document 7] Agaugue et al. Blood. 2011, 117: 7021-7031 [Non-Patent Document 8] Colonna et al. J Immunol. 1998, 160: 3096-3100 [Non-Patent Document 9] Allan et al. J Exp Med. 1999, 189(7): 1149-1155 [Non-Patent Document 10] Liang et al. Proc Natl Sci USA. 2008, 105(24): 8357 [Non-Patent Document 11] Ristich et al. Eur J Immunol. 2005, 35: 1133-1142 Summary of the Invention [Problem to be solved by the invention]

[0008] As a result, there is a need for stable co-formulations of anti-ILT4 and anti-PD-1 antibodies, preferably exhibiting stability over months to years under conditions typical of storage of drugs for self-administration, i.e., in a syringe at refrigerator temperatures, resulting in an extended shelf life for the corresponding pharmaceutical product. [Means for solving the problem]

[0009] In one aspect, the invention provides a co-formulation of an anti-human ILT4 (anti-ILT4) antibody or antigen-binding fragment thereof with an anti-PD-1 antibody or antigen-binding fragment thereof. In one formulation, the co-formulation comprises: (i) about 10 mg / mL to about 200 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mg / mL to about 200 mg / mL of an anti-PD-1 antibody or antigen-binding fragment thereof; (iii) about 5 mM to about 20 mM of a buffer; (iv) about 6% to about 8% weight / volume (w / v) of a non-reducing sugar; (v) about 0.01% to about 0.10% (w / v) of a non-ionic surfactant; and (vi) about 1 mM to about 20 mM of an antioxidant; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and and CDR-L3: QSFDNSLSAYV (SEQ ID NO:21); wherein the anti-PD-1 antibody, or antigen-binding fragment thereof, comprises a light chain variable domain comprising a CDRL1 of SEQ ID NO:195, a CDRL2 of SEQ ID NO:296, and a CDRL3 of SEQ ID NO:397. and a variable heavy chain region comprising a CDRH1 of SEQ ID NO: 6100, a CDRH2 of SEQ ID NO: 7101, and a CDRH3 of SEQ ID NO: 8102; wherein the buffer is an L-histidine buffer, an acetate buffer, or a citrate buffer, wherein the non-reducing sugar is a disaccharide, wherein the non-ionic surfactant is polysorbate 20 or polysorbate 80, and wherein the antioxidant is methionine.

[0010] In some embodiments, the non-reducing sugar is sucrose. In some embodiments, the buffer is an L-histidine buffer. In some embodiments, the buffer is an L-histidine buffer at a pH of about 5.0 to 6.5. In certain embodiments, the non-ionic surfactant is polysorbate 80 (PS-80). In some embodiments, the antioxidant is L-methionine.

[0011] Accordingly, in another aspect, provided herein is a formulation of an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 10 mg / mL to about 200 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mg / mL to about 200 mg / mL of an anti-PD-1 antibody or antigen-binding fragment thereof; (iii) about 5 mM to about 20 mM L-histidine buffer; (iv) about 6% to about 8% (w / v) sucrose; (v) about 0.01% to about 0.10% (w / v) PS-80; and (vi) about 1 mM to about 20 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof has the following structure: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A). and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO:19), CDR-L2: GX1X2NRPS, where X1 is N, Q, E, or D, and X2 is S or A (SEQ ID NO:20) and CDR-L3: QSFDNSLSAYV (SEQ ID NO:21); and wherein the anti-PD-1 antibody, or antigen-binding fragment thereof, comprises a variable light chain region comprising a CDRL1 of SEQ ID NO:95, a CDRL2 of SEQ ID NO:96, and a CDRL3 of SEQ ID NO:97; and a variable heavy chain region comprising a CDRH1 of SEQ ID NO:100, a CDRH2 of SEQ ID NO:101, and a CDRH3 of SEQ ID NO:102.

[0012] In some embodiments, the formulation comprises about 8 mM to about 12 mM L-histidine buffer.

[0013] In certain embodiments, the formulation comprises about 5 mM to about 10 mM L-methionine.

[0014] In another embodiment, the formulation comprises about 0.01% to about 0.05% (w / v) PS-80.

[0015] In yet another embodiment, the formulation comprises about 10 mg / mL to about 150 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 10 mg / mL, about 12.5 mg / mL, about 15 mg / mL, about 25 mg / mL, about 50 mg / mL, about 75 mg / mL, about 100 mg / mL, about 125 mg / mL or about 150 mg / mL. In one embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 10 mg / mL. In another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 12.5 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 15 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 25 mg / mL. In one embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 50 mg / mL. In another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 75 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 100 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 125 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 150 mg / mL.

[0016] Thus, in one particular embodiment, the formulation comprises about 25 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0017] In another specific embodiment, the formulation comprises about 50 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0018] In yet another specific embodiment, the formulation comprises about 75 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0019] In yet another specific embodiment, the formulation comprises about 100 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0020] In yet another specific embodiment, the formulation comprises about 125 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0021] In certain embodiments of the various formulations provided herein, the formulation has a pH range of about 5.0 to about 6.8. In some embodiments, the formulation has a pH range of about 5.5 to about 6.0. In other embodiments, the formulation has a pH of about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, or about 6.0. In one embodiment, the formulation has a pH of about 5.5. In another embodiment, the formulation has a pH of about 5.6. In yet another embodiment, the formulation has a pH of about 5.7. In yet another embodiment, the formulation has a pH of about 5.8. In another embodiment, the formulation has a pH of about 5.9. In yet another embodiment, the formulation has a pH of about 6.0.

[0022] Thus, in one particular embodiment, the pharmaceutical formulation of an anti-ILT4 antibody or antigen-binding fragment thereof comprises: (i) about 50 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mg / mL to about 200 mg / mL of an anti-PD-1 antibody or antigen-binding fragment thereof; (iii) about 10 mM L-histidine buffer, pH about 5.5; (iv) about 7% (w / v) sucrose; (v) about 0.025% (w / v) polysorbate 80; and (vi) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof has the structure of CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17). and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO:19), CDR-L2: GX1X2NRPS, where X1 is N, Q, E, or D, and X2 is S or A (SEQ ID NO:20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO:21), and wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises a variable light chain region comprising a CDRL1 of SEQ ID NO:95, a CDRL2 of SEQ ID NO:96, and a CDRL3 of SEQ ID NO:97; and a variable heavy chain region comprising a CDRH1 of SEQ ID NO:100, a CDRH2 of SEQ ID NO:101, and a CDRH3 of SEQ ID NO:10.

[0023] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E, or D and X2 is S or A) (SEQ ID NO: 20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0024] In some embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0025] In another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57, and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:58.

[0026] In yet another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2, and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0027] In yet another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:80, and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0028] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:57, and a light chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:58.

[0029] In some embodiments of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:2, and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7.

[0030] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof is a monoclonal antibody comprising two heavy chains comprising the amino acid sequence set forth in SEQ ID NO:2 and two light chains comprising the amino acid sequence set forth in SEQ ID NO:7.

[0031] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof is a monoclonal antibody comprising two heavy chains consisting of the amino acid sequences set forth in SEQ ID NO:2 and two light chains consisting of the amino acid sequences set forth in SEQ ID NO:7.

[0032] In another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:80, and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7.

[0033] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof is a monoclonal antibody comprising two heavy chains comprising the amino acid sequence set forth in SEQ ID NO:80 and two light chains comprising the amino acid sequence set forth in SEQ ID NO:7.

[0034] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof is a monoclonal antibody comprising two heavy chains consisting of the amino acid sequences set forth in SEQ ID NO:80 and two light chains consisting of the amino acid sequences set forth in SEQ ID NO:7.

[0035] In one embodiment, the formulation is a liquid formulation. In one embodiment, the formulation is a frozen formulation. In another embodiment, the liquid formulation is stored at about 5° C. In yet another embodiment, the formulation is stored frozen at about −20° C. or below. In yet another embodiment, the formulation is stored frozen at about −70° C. or below. In yet another embodiment, the liquid formulation is a solution reconstituted from a lyophilized formulation.

[0036] In certain embodiments, after storage of the formulation at about 3° C. to about 5° C. (e.g., 3° ​​C., 4° C., 5° C.) for up to 9 months, (i) the % monomer of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 99% as measured by ultra-performance size exclusion chromatography; (ii) the % monomer of the anti-PD-1 antibody or antigen-binding fragment thereof is at least about 98% as measured by ultra-performance size exclusion chromatography; (iii) the OD 350-500 (iv) the turbidity of the formulation is at most about 0.135 when measured by high performance ion exchange chromatography; (v) the % main peak of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 52%, the % acidic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 15%, and / or the % basic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 12%, when measured by high performance ion exchange chromatography; (v) the % main peak of the anti-PD-1 antibody or antigen-binding fragment thereof is at least about 10%, and the turbidity of the formulation is at most about 0.135 when measured by high performance ion exchange chromatography; or the % acidic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 5%, and / or the % basic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 4%; (vi) the number of subvisible particles of particles at least 2 μm in size is at most about 3500, as measured by microflow imaging; and / or (vii) the % oxidation of one or more amino acid residues selected from the group consisting of W7, W102, M253, M359 and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4%, as measured by reduced peptide mapping.

[0037] In some embodiments, the % monomer of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 99% when measured by ultra performance size exclusion chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months.

[0038] In some embodiments, the formulation is stored at about 3° C. to about 5° C. for 6 months and then the OD 350-500 When measured at 100° C., the turbidity of the formulation is a maximum of about 0.135.

[0039] In some embodiments, the % main peak of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 63% when measured by high performance ion exchange chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months. In some embodiments, the % acidic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 23% when measured by high performance ion exchange chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months. In some embodiments, the % basic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 14% when measured by high performance ion exchange chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months. In some embodiments, after storage of the formulation at about 3° C. to about 5° C. for 6 months, the % main peak of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 63%, the % acidic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 23%, and the % basic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 14% when measured by high performance ion exchange chromatography.

[0040] In some embodiments, after storage of the formulation at about 3° C. to about 5° C. for 6 months, the number of sub-visible particles having a size of at least 2 μm is up to about 3500 as measured by microflow imaging.

[0041] In some embodiments, the percent oxidation of one or more amino acid residues selected from the group consisting of W7, W102, M253, M359 and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% as measured by reduced peptide mapping after storage of the formulation at about 3° C. to about 5° C. for 6 months. In one embodiment, the amino acid residue is W7 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80. In another embodiment, the amino acid residue is W102 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80. In yet another embodiment, the amino acid residue is M253 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80. In yet another embodiment, the amino acid residue is M359 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80. In yet another embodiment, the amino acid residue is M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80.

[0042] In one embodiment, the % oxidation rate of one amino acid residue selected from the group consisting of W7, W102, M253, M359 and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4% when measured by reduced peptide mapping after storage of the formulation at about 3° C. to about 5° C. for 6 months. In one embodiment, the % oxidation rate of two amino acid residues selected from the group consisting of W7, W102, M253, M359 and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4% when measured by reduced peptide mapping after storage of the formulation at about 3° C. to about 5° C. for 6 months. In one embodiment, when the formulation is stored at about 3° C. to about 5° C. for 6 months and then measured by reduced peptide mapping, the % oxidation rate of three amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4%. In one embodiment, when the formulation is stored at about 3° C. to about 5° C. for 6 months and then measured by reduced peptide mapping, the % oxidation rate of four amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4%. In one embodiment, the % oxidation of all five amino acid residues from the group consisting of W7, W102, M253, M359 and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% when measured by reduced peptide mapping after storage of the formulation at about 3°C ​​to about 5°C for 6 months.

[0043] In yet another aspect, there is provided herein a method of treating cancer in a human patient in need of such treatment, wherein the method comprises administering a therapeutically effective amount of a pharmaceutical formulation described herein.

[0044] In yet another aspect, there is provided herein a use of a pharmaceutical formulation described herein for the preparation of a medicament for treating cancer. [Brief description of the drawings]

[0045] [Figure 1] FIG. 1 shows high molecular weight (HMW) species measured by UP-SEC in anti-ILT4 / anti-PD-1 co-formulation, anti-ILT4 SE, and pembrolizumab SE samples at various concentrations of L-methionine and with various levels of light exposure and residual metals, as described in Example 6. [Diagram 2] FIG. 2 shows PS80 content measured by reverse phase chromatography in anti-ILT4 / anti-PD-1 co-formulation, anti-ILT4 SE, and pembrolizumab SE samples at various concentrations of L-methionine and with various levels of light exposure and residual metals, as described in Example 6. [Diagram 3] FIG. 3 shows the oxidation of M105 as a function of metal concentration and light exposure as measured by reduced peptide mapping for anti-ILT4 / anti-PD-1 co-formulation samples with various concentrations of L-methionine (7-15 mM) at 40° C. for 4 weeks, as described in Example 6. [Figure 4] FIG. 4 shows the oxidation of W102 as a function of metal concentration and light exposure as measured by reduced peptide mapping for anti-ILT4 / anti-PD-1 co-formulations and anti-ILT4 SE samples with various concentrations of L-methionine (7-15 mM) at 40° C. for 4 weeks, as described in Example 6. [Diagram 5] FIG. 5 shows the oxidation of W7 as a function of metal concentration and light exposure as measured by reduced peptide mapping for anti-ILT4 / anti-PD-1 co-formulations and anti-ILT4 SE samples with various concentrations of L-methionine (7-15 mM) at 40° C. for 4 weeks, as described in Example 6. [Figure 6] FIG. 6 shows the total oxidation of methionine residues as a function of metal concentration and light exposure as measured by hydrophobic interaction chromatography for pembrolizumab SE samples with various concentrations of L-methionine (7-15 mM) at 40° C. for 4 weeks, as described in Example 6. [Figure 7]FIG. 7 shows contour plots of predicted values ​​of HMW co-formulations at various factor levels at 40° C., as described in Example 7. [Figure 8] FIG. 8 shows a contour plot of HMW predicted values ​​at various factor levels for anti-ILT4 SE at 40° C. as described in Example 7. [Figure 9] FIG. 9 shows a contour plot of predicted HMW values ​​at various factor levels for pembrolizumab SE at 40° C., as described in Example 7. [Figure 10] FIG. 10 shows a contour plot of predicted % acidic variants of anti-ILT4 in co-formulations at various factor levels at 40° C. for 4 weeks, as described in Example 7. [Figure 11] 11 shows a contour plot of % major component for anti-ILT4 in co-formulations as a function of various factors (L-methionine concentration, light exposure, and metal content). The figure shows that with increasing light stress, as described in Example 7, the % major component of anti-ILT4 decreases. [Figure 12] 12 shows a contour plot of % basic variants for anti-ILT4 in co-formulations as a function of various factors (L-methionine concentration, light exposure, and metal content). The figure shows that increasing light stress increases the % basic variants of anti-ILT4, as described in Example 7. [Figure 13] FIG. 13 shows contour plots of predicted values ​​over time from model fits across different settings of three parameters (L-methionine concentration, light exposure, metal content), as described in Example 7. [Figure 14] FIG. 14 shows a contour plot of % major component for pembrolizumab in co-formulations as a function of various factors (L-methionine concentration, light exposure, and metal content), as described in Example 7. [Figure 15]FIG. 15 shows a contour plot of % basic variant for pembrolizumab in co-formulations as a function of various factors (L-methionine concentration, light exposure, and metal content), as described in Example 7. [Figure 16] FIG. 16 shows contour plots of predicted values ​​over time from model fits across different settings of three parameters (L-methionine concentration, light exposure, metal content), as described in Example 7. [Figure 17] FIG. 17 shows a contour plot of % major component for anti-ILT4 in anti-ILT4 SE formulations as a function of various factors (L-methionine concentration, light exposure and metal content) as described in Example 7. [Figure 18] FIG. 18 shows a contour plot of % basic variant for anti-ILT4 in anti-ILT4 SE formulation as a function of various factors (L-methionine concentration, light exposure and metal content) as described in Example 7. [Figure 19] FIG. 19 shows the L-methionine concentrations at all stability time points for all three formulations, as described in Example 7. [Figure 20] FIG. 20 shows a plot of turbidity values ​​for all three formulations under conditions of 5° C. and 40° C., as described in Example 8. [Figure 21] Figures 21-25 show raw data from peptide mapping for SE formulations and co-formulations under initial and 40°C conditions as described in Example 8. Figure 21 shows M253 (shared), Figure 22 shows M429 (shared), Figure 23 shows W7 (anti-ILT4), Figure 24 shows W102 (anti-ILT4), and Figure 25 shows M105 (pembrolizumab). [Figure 22]Figures 21-25 show raw data from peptide mapping for SE formulations and co-formulations under initial and 40°C conditions as described in Example 8. Figure 21 shows M253 (shared), Figure 22 shows M429 (shared), Figure 23 shows W7 (anti-ILT4), Figure 24 shows W102 (anti-ILT4), and Figure 25 shows M105 (pembrolizumab). [Diagram 23] Figures 21-25 show raw data from peptide mapping for SE formulations and co-formulations under initial and 40°C conditions as described in Example 8. Figure 21 shows M253 (shared), Figure 22 shows M429 (shared), Figure 23 shows W7 (anti-ILT4), Figure 24 shows W102 (anti-ILT4), and Figure 25 shows M105 (pembrolizumab). [Figure 24] Figures 21-25 show raw data from peptide mapping for SE formulations and co-formulations under initial and 40°C conditions as described in Example 8. Figure 21 shows M253 (shared), Figure 22 shows M429 (shared), Figure 23 shows W7 (anti-ILT4), Figure 24 shows W102 (anti-ILT4), and Figure 25 shows M105 (pembrolizumab). [Diagram 25] Figures 21-25 show raw data from peptide mapping for SE formulations and co-formulations under initial and 40°C conditions as described in Example 8. Figure 21 shows M253 (shared), Figure 22 shows M429 (shared), Figure 23 shows W7 (anti-ILT4), Figure 24 shows W102 (anti-ILT4), and Figure 25 shows M105 (pembrolizumab). [Figure 26] FIG. 26 shows high molecular weight (HMW) and low molecular weight (LMW) species as measured by UP-SEC in co-formulation 1 (CF1) and co-formulation 2 (CF2) when stored for 12 months at 5° C. (circles), 6 months at 25° C. (upward triangles), and 6 months at 40° C. (downward triangles), as described in Example 8. [Figure 27] FIG. 27 shows the charge variants of anti-ILT4 and pembrolizumab as measured by IEX in CF1 and CF2 when stored for 12 months at 5° C. (circles), 6 months at 25° C. (upward triangles), and 6 months at 40° C. (downward triangles), as described in Example 8. [Figure 28] FIG. 28 shows the total L-methionine content in CF1 and CF2 when stored for 12 months at 5° C. (circles), 6 months at 25° C. (downward triangles), and 6 months at 40° C. (upward triangles), as described in Example 8. [Figure 29] FIG. 29 shows the total PS-80 content in CF1 and CF2 when stored for 12 months at 5° C. (circles), 6 months at 25° C. (upward triangles), and 6 months at 40° C. (downward triangles), as described in Example 8. [Diagram 30] FIG. 30 shows the number of subvisible particles in CF1 and CF2 when stored for 12 months at 5° C. (light grey), 6 months at 25° C. (dark grey), and 6 months at 40° C. (black), as described in Example 8. [Diagram 31] FIG 31 shows the oxidation of M105 (anti-PD-1), M429 (shared residue) and M253 (shared residue) as measured by reduced peptide mapping CF1 and CF2 upon storage for 12 months at 5°C (circles), 6 months at 25°C (up triangles) and 6 months at 40°C (down triangles) as described in Example 8. [Diagram 32] FIG. 32 shows the oxidation of W102 (anti-ILT4 monoclonal antibody) and W7 (anti-ILT4 monoclonal antibody) measured by reduced peptide mapping CF1 and CF2 when stored for 12 months at 5° C. (circles), 6 months at 25° C. (upward triangles) and 6 months at 40° C. (downward triangles) as described in Example 8. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0046] Definitions and Abbreviations As used throughout this specification and the appended claims, the following abbreviations apply: [Table 1]

[0047] Unless clearly defined elsewhere in this specification, all technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this invention belongs.

[0048] As used throughout this specification and in the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0049] References to "or" indicate either or both possibilities, unless the context clearly dictates otherwise. In some cases, "and / or" is used to emphasize either or both possibilities.

[0050] As used herein, "acidic variant" refers to an anti-ILT4 antibody that is more acidic (e.g., as determined by cation exchange chromatography) compared to the major species of the anti-ILT4 antibody. Such acidic variants are detected by various chromatographic purification methods to separate molecular variants by charge, e.g., ion exchange chromatography, e.g., cation exchange chromatography or WCX-10 HPLC (weak cation exchange chromatography), and optionally subsequently detected by mass spectrometry. In general, acidic variants have a lower isoelectric point (pI) than the major species and may be more acidic in nature, e.g., due to methionine oxidation, sialylation of asparagine residues, or deamidation variants of the antibody, or a combination thereof. In one embodiment, the anti-ILT4 antibody acidic variant is the anti-ILT4 antibody species identified by the acidic variant peak in Figure 6D and eluted according to the cation ion exchange method described in Example 11. In the ion exchange chromatography method, "% acidic variant species" refers to the total area of ​​the acidic variant peak divided by the total area of ​​all peaks in the elution chromatogram. In one embodiment, the anti-ILT4 antibody acidic variant is identified by a peak eluting before the main peak according to a cationic ion exchange method. In another embodiment, the anti-ILT4 antibody acidic variant is identified by a peak eluting before the main peak according to a weak cationic ion exchange method. In an ion exchange chromatography method, "% acidic variant" refers to the total area of ​​the acidic species peak divided by the total area of ​​all peaks in the elution chromatogram.

[0051] "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y is generally measured by the dissociation constant (K DAffinity can be measured by common methods known in the art, such as KinExA and Biacore.

[0052] As used herein, the term "antibody" includes, but is not limited to, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), fully human antibodies, and chimeric antibodies.

[0053] As used herein, unless otherwise indicated, "antigen-binding fragment" refers to an antigen-binding fragment of an antibody, i.e., an antibody fragment that retains the ability to bind to the antigen bound by the full-length antibody (e.g., a fragment that retains one or more CDR regions). Examples of antibody-binding fragments include, but are not limited to, the following: Fab, Fab', F(ab') 2 , Fv fragments, and individual antibody heavy or light chains, and individual heavy or light chain variable regions.

[0054] As used herein, "basic variant" refers to an anti-ILT4 antibody that is more basic (e.g., as determined by cation exchange chromatography) compared to the major species of the anti-ILT4 antibody. Such basic variants are detected by various chromatographic purification methods to separate molecular variants by charge, for example, ion exchange chromatography, for example, cation exchange chromatography (e.g., the method described in Example 11) or WCX-10 HPLC (weak cation exchange chromatography), and optionally subsequently detected by mass spectrometry. In general, basic variants have a higher pH than the major species and may be more acidic in nature due to modifications or differences from the major species. In one embodiment, the anti-ILT4 antibody basic species is identified by a peak that elutes after the major peak according to a cation ion exchange method. In another embodiment, the anti-ILT4 antibody basic species is identified by a peak that elutes after the major peak according to a weak cation ion exchange method. In an ion exchange chromatography method, "% basic species" refers to the total area of ​​the basic species peak divided by the total area of ​​all peaks in the elution chromatogram.

[0055] A "Fab fragment" is composed of one light chain and the CH1 and variable regions of one heavy chain. The heavy chain of a Fab molecule cannot form disulfide bonds with another heavy chain molecule. A "Fab fragment" can be the product of papain cleavage of an antibody.

[0056] The "Fc" region comprises two heavy chain fragments comprising the CH1 and CH2 domains of an antibody, held together by two or more disulfide bonds and by hydrophobic interactions of the CH3 domain.

[0057] A "Fab' fragment" is a fragment that contains one light chain and one V Hand a portion or fragment of one heavy chain that includes the CH1 domain and the CH2 domain and further includes the region between the CH1 domain and the CH2 domain, whereby interchain disulfide bonds are formed between the two heavy chains of the two Fab' fragments to form F(ab'). 2 Molecules can be formed.

[0058] "F(ab') 2 A "F(ab')" fragment contains two light chains and two heavy chains that contain a portion of the constant region between the CH1 and CH2 domains, thereby forming an interchain disulfide bond between the two heavy chains. 2 The "F(ab')" fragment is thus composed of two Fab' fragments held together by disulfide bonds between the two heavy chains. 2 "Fragments" may be the products of pepsin cleavage of antibodies.

[0059] The "Fv region" comprises the variable regions from both the heavy and light chains, but lacks the constant regions.

[0060] "Isolated antibody" refers to a purified state, and in this context means that the molecule is substantially free of other biological molecules (e.g., nucleic acids, proteins, lipids, carbohydrates) or other substances (e.g., cell debris and growth medium). In general, the term "isolated" is not intended to imply the complete absence of such substances, or the absence of water, buffers or salts, unless such substances or water, buffers or salts are present in an amount that would substantially interfere with the experimental or therapeutic use of the binding compounds described herein.

[0061] As used herein, "major species" or "major" refers to an anti-ILT4 antibody species identified as the majority of antibody species in a mixture with one or more acidic or basic species. Such a major species is detected by various chromatographic purification methods to separate molecular variants by charge, for example, by ion exchange chromatography, for example, cation exchange chromatography (e.g., the method described in Example 11) or WCX-10 HPLC (weak cation exchange chromatography), and optionally subsequently detected by mass spectrometry. The mixture may be, for example, the result of an antibody preparation from mammalian cells and its post-translational modification, upstream and downstream processing or storage. In one embodiment, the major species is identified as the major peak according to a cation ion exchange method. In the ion exchange method, "% major" refers to the total area of ​​the major peak divided by the total area of ​​all peaks in the elution chromatogram. In one embodiment to measure the major species, acidic species or basic species, a Thermo Scientific ProPac WCX-10 column is used for the cation ion exchange method. In another embodiment, a Thermo Scientific ProPac WCX-10 column is used, where the mobile phase (A) is 24 mM MES pH 6.1 (containing 4% acetonitrile), the mobile phase (B) is 20 mM sodium phosphate, 95 mM NaCl pH 8.0 (containing 4% acetonitrile), and the column temperature is 35° C. In one embodiment, the following non-linear gradient is used: 0-0.6 min 22%-22% B; 0.6-15.0 min 22%-29% B; 15.0-30.0 min 29%-70% B; 30.0-30.5 min 70%-100% B; and 30.5-33.0 min 100%-100% B. In a further embodiment, the cation ion exchange method is described in Example 11.

[0062] The term "monoclonal antibody" as used herein refers to a population of substantially homogeneous antibodies, i.e., the antibody molecules comprising the population are identical in amino acid sequence, except for possible naturally occurring mutations that may be present in minor amounts. In contrast, conventional (polyclonal) antibody preparations typically contain a large number of different antibodies having different amino acid sequences in the variable domains, often specific for different epitopes. The modifier "monoclonal" indicates the nature of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler et al. ((1975) Nature 256: 495), or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). The monoclonal antibodies can also be isolated from phage antibody libraries using the techniques described in, for example, Clackson et al. (1991) Nature 352: 624-628 and Marks et al. (1991) J. Mol. Biol. 222: 581-597. See also Presta (2005) J. Allergy Clin. Immunol. 116:731.

[0063] The terms "fully human antibody" or "human antibody" refer to an antibody that contains only human immunoglobulin protein sequences. A fully human antibody may contain mouse carbohydrate chains if produced in a mouse, in a mouse cell, or in a hybridoma derived from a mouse cell. Similarly, a "mouse antibody" refers to an antibody that contains only mouse immunoglobulin sequences. Alternatively, a fully human antibody may contain rat carbohydrate chains if produced in a rat, in a rat cell, or in a hybridoma derived from a rat cell. Similarly, a "rat antibody" refers to an antibody that contains only rat immunoglobulin sequences.

[0064] Generally, the basic "antibody" structural unit comprises a tetramer. In monospecific antibodies, each tetramer comprises two pairs of identical polypeptide chains, each pair having one "light chain" (about 25 kDa) and one "heavy chain" (about 50-70 kDa). The amino-terminal portion of each chain contains a "variable region" or "variable domain" of about 100-110 or more amino acids that are primarily responsible for antigen recognition. The carboxy-terminal portion of the heavy chain may define a constant region that is primarily responsible for effector function.

[0065] Typically, human constant light chains are classified as kappa and lambda light chains. Furthermore, human constant heavy chains are typically classified as mu, delta, gamma, alpha or epsilon, defining the antibody's isotype as IgM, IgD, IgG, IgA and IgE, respectively. These IgG subtypes include, for example, IgG1 and IgG4. The present invention encompasses anti-ILT4 antibodies and antigen-binding fragments comprising any of these light and / or heavy constant chains.

[0066] As used herein, "variable region," "variable domain," "V region," or "V chain" refers to the segment of an IgG chain that is variable in sequence among different antibodies. The "variable region" of an antibody refers to the variable region of the antibody light chain or the variable region of the antibody heavy chain, either alone or in combination. The variable region of the heavy chain is referred to as the "V H The variable region of the light chain may be referred to as "V L". Typically, both heavy and light chain variable regions contain three hypervariable regions (also called complementarity determining regions (CDRs)) located within relatively conserved framework regions (FRs). The CDRs are usually aligned with the framework regions and enable binding to a specific epitope. Generally, from N-terminus to C-terminus, both light and heavy chain variable domains contain FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. The assignment of amino acids to each domain generally follows the definitions in "Sequences of Proteins of Immunological Interest, Kabat, et al.; National Institutes of Health, Bethesda, Md.; 5th ed.; NIH Publ. No. 91-3242 (1991)," "Kabat (1978) Adv. Prot. Chem. 32:1-75," "Kabat, et al., (1977) J. Biol. Chem. 252:6609-6616," "Chothia, et al., (1987) J Mol. Biol. 196:901-917," or "Chothia, et al., (1989) Nature 342:878-883."

[0067] "CDR" is the antibody V H One of the three hypervariable regions (H1, H2, or H3) within the non-framework regions of the β-sheet framework, or LIt indicates one of three hypervariable regions (L1, L2, or L3) in the non-framework regions of the β-sheet framework. Thus, CDRs are variable region sequences interspersed within the framework region sequences. CDR regions are well known to those skilled in the art and are defined, for example, by Kabat as the most hypervariable regions in antibody variable domains. CDR region sequences are also structurally defined by Chothia as residues that are not part of the conserved β-sheet framework and therefore can adapt to various conformations. Both terms are well recognized in the art. CDR region sequences are also defined by AbM, Contact, and IMGT. The positions of CDRs in canonical antibody variable regions have been determined by comparison of multiple structures (Al-Lazikani et al., 1997, J. Mol. Biol. 273:927-48; Morea et al., 2000, Methods 20:267-79). Since the number of residues in the hypervariable regions varies among antibodies, additional residues relative to the canonical position are conventionally numbered a, b, c, etc., next to the residue number in the canonical variable region numbering scheme (Al-Lazikani et al., supra). Such nomenclature is likewise familiar to those of skill in the art. Correspondences between numbering systems, including, for example, Kabat numbering and the IMGT specific numbering system, are familiar to those of skill in the art and are shown in Table 1 below. In some embodiments, the CDRs are defined by the Kabat numbering system. In other embodiments, the CDRs are defined by the IMGT numbering system. In yet other embodiments, the CDRs are defined by the AbM numbering system. In yet other embodiments, the CDRs are defined by the Chothia numbering system. In yet other embodiments, the CDRs are defined by the Contact numbering system.

[0068] [Table 2]

[0069] Sequence identity indicates the degree to which the amino acids of two polypeptides are identical in equal positions when the two sequences are optimally aligned.

[0070] Sequence similarity includes identical residues and non-identical amino acids that are biochemically related. Biochemically related amino acids that share similar properties and may be interchangeable are discussed above.

[0071] "Conservatively modified variants" or "conservative substitutions" refer to the substitution of amino acids in a protein with other amino acids having similar properties (e.g., charge, side chain size, hydrophobicity / hydrophilicity, backbone conformation and rigidity, etc.), such that such changes can often be made without altering the biological activity of the protein. Those skilled in the art will generally recognize that single amino acid substitutions in non-essential regions of a polypeptide do not substantially alter biological activity (see, e.g., Watson et al. (1987) Molecular Biology of the Gene, The Benjamin / Cummings Pub. Co., p. 224 (4th Ed.)). In addition, substitutions of structurally or functionally similar amino acids are less likely to destroy biological activity. Exemplary conservative substitutions are shown in Table 2 below.

[0072] [Table 3]

[0073] As used herein, the term "epitope" refers to the area or region on an antigen to which an antibody or antigen-binding fragment binds. Binding of an antibody or antigen-binding fragment thereof disclosed herein to an epitope means that the antibody or antigen-binding fragment thereof binds to one or more amino acid residues within the epitope.

[0074] As used herein, "treat" or "treating" a cancer or infectious condition means administering a formulation of the invention to a subject having an immune or cancerous condition or diagnosed with cancer or a pathogenic infection (e.g., viral, bacterial, fungal) to achieve at least one positive therapeutic effect (e.g., a reduction in the number of cancer cells, a reduction in tumor size, a reduction in the rate of cancer cell invasion into peripheral organs, or a reduction in the rate of tumor metastasis or growth, etc.). "Treatment" may include one or more of the following: inducing / increasing an anti-tumor immune response, stimulating an immune response to pathogens, toxins and / or self-antigens, stimulating an immune response to viral infection, reducing the number of one or more tumor markers, inhibiting tumor cell proliferation or survival, eliminating or reducing the size of one or more cancerous lesions or tumors, reducing the level of one or more tumor markers, ameliorating cancer, reducing the severity or duration of cancer, prolonging patient survival compared to the expected survival of a similar untreated patient.

[0075] The terms "cancer", "cancerous" or "malignant" refer to or describe a physiological condition in mammals that is typically characterized by unregulated proliferation of cells. Examples of cancer include, but are not limited to, carcinoma, lymphoma, leukemia, blastoma and sarcoma. More specific examples of such cancers include squamous cell carcinoma, myeloma, small cell lung cancer, non-small cell lung cancer, glioma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, gastrointestinal (ductal) cancer, renal cancer, ovarian cancer, liver cancer, lymphoblastic leukemia, lymphocytic leukemia, colon cancer, endometrial cancer, kidney cancer, prostate cancer, thyroid cancer, melanoma, chondrosarcoma, neuroblastoma, pancreatic cancer, glioblastoma multiforme, cervical cancer, brain tumor, stomach cancer, bladder cancer, liver cancer, breast cancer, colon cancer and head and neck cancer.

[0076] The term "patient" (alternatively referred to herein as "subject" or "individual") refers to a mammal (e.g., rat, mouse, dog, cat, rabbit) that can be treated with the formulations of the invention, and most preferably a human. In some embodiments, the patient is an adult patient. In other embodiments, the patient is a pediatric patient. A patient "in need of treatment" is an individual who has been diagnosed with, is suspected of being, or is susceptible to a disease or disorder that the formulations of the invention are intended to treat or ameliorate (e.g., an ILT4-associated disease, e.g., cancer), or in whom it is desired to prevent such a disorder.

[0077] The term "therapeutically effective amount" or "effective amount" refers to the amount of a therapeutic composition or formulation introduced into a patient that is sufficient to treat a disease or condition. One of skill in the art will recognize that this level may vary depending on patient characteristics such as age, weight, etc.

[0078] The term "about," when modifying an amount of a substance or composition (e.g., mM or M), a proportion of a formulation component (v / v or w / v), the pH of a solution / formulation, or the value of a parameter characterizing a step of a method, refers to a variation in a numerical quantity that may occur, for example, through typical measuring, handling, and sampling procedures associated with preparing, characterizing, and / or using a substance or composition; through instrumental errors in these procedures; through differences in the manufacture, source, or purity of ingredients employed in making or using a composition or performing a procedure; etc. In certain embodiments, "about" may mean a variation of ±0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, or 10%.

[0079] As used herein, "x% (w / v)" corresponds to x g / 100 mL (e.g., 5% w / v corresponds to 50 mg / mL).

[0080] As used throughout this specification and claims, the phrase "consists essentially of" or variations such as "consist essentially of" or "consisting essentially of" refers to the inclusion of any recited element or group of elements, and optionally the inclusion of other elements of a similar or different nature to the recited elements, that do not substantially alter the basic or novel characteristics of the specified dosing regimen, method, or composition. As a non-limiting example, a binding compound consisting essentially of a recited amino acid sequence may contain one or more amino acids that include substitutions of one or more amino acid residues that do not substantially affect the properties of the binding compound.

[0081] The word "comprising" or variations thereof, such as "comprise" or "comprises," is used throughout this specification and claims in its inclusive sense, i.e., unless the context requires otherwise, either by express language or by necessary implication, to specify the presence of stated features but not to exclude the presence or addition of further features that may materially enhance the practice or utility of any of the embodiments of the invention.

[0082] The term "buffering agent" includes agents that maintain the pH of a solution of the formulation within an acceptable range, or, in the case of a lyophilized formulation of the invention, provides an acceptable pH of the solution prior to lyophilization.

[0083] The terms "lyophilization," "lyophilized," and "freeze-dried" refer to a process in which the material to be dried is first frozen and then the ice or frozen solvent is removed by sublimation under reduced pressure. Excipients can be included in the pre-lyophilized formulation to enhance the storage stability of the lyophilized product.

[0084] The term "pharmaceutical formulation" refers to a preparation in a form that allows the active ingredient to be effective and that does not contain additional ingredients that are toxic to the subject to which it is administered. The terms "formulation" and "pharmaceutical formulation" are used interchangeably throughout.

[0085] "Pharmaceutically acceptable" refers to excipients (vehicles, additives) and compositions that can be reasonably administered to a subject to provide an effective dose of the active ingredient employed, that are "generally regarded as safe," and that, for example, are physiologically tolerated when administered to humans and do not typically produce an allergic reaction or similar unpleasant reaction (e.g., stomach upset). In another embodiment, the term refers to molecular entities and compositions approved by a federal or state government regulatory agency or listed in the United States Pharmacopeia or other generally recognized pharmacopoeias for use in animals (more particularly, humans).

[0086] A "reconstituted" formulation is one in which a lyophilized protein formulation has been dissolved in a diluent and prepared such that the protein is dispersed in the reconstituted formulation. The reconstituted formulation is suitable for administration (e.g., parenteral administration), and optionally, subcutaneous administration.

[0087] A "stable" formulation is one in which the protein therein essentially retains its physical and / or chemical stability and / or biological activity under storage or stress conditions. Various analytical techniques for measuring protein stability are available in the art and are reviewed in Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, NY, Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10:29-90 (1993). Stability may be measured at a selected temperature for a selected period of time. For example, in one embodiment, a stable formulation is one in which no significant change is observed at refrigerated temperatures (2-8°C) for at least 6 months. In one embodiment, a stable formulation is one in which no significant change is observed at refrigerated temperatures (2-8°C) for at least 12 months. In another embodiment, a stable formulation is one in which no significant change is observed at refrigerated temperatures (2-8°C) for at least 24 months. In another embodiment, a stable formulation is one in which no significant change is observed at room temperature (23-27°C) for at least 3 months. In another embodiment, a stable formulation is one in which no significant change is observed at room temperature (23-27°C) for at least 6 months. In another embodiment, a stable formulation is one in which no significant change is observed at room temperature (23-27°C) for at least 12 months. In another embodiment, a stable formulation is one in which no significant change is observed at room temperature (23-27°C) for at least 18 months. Stability criteria for antibody formulations are as follows. Typically, the degradation of antibody monomer is 10% or less, preferably 5% or less, as measured by SEC-HPLC. Typically, the formulation is colorless or clear to slightly opaque upon visual analysis. Typically, the concentration, pH, and osmolality of the formulation vary within ±10%. Titers are typically within 60-140%, preferably 80-120%, of a control or reference. Typically, antibody clipping observed is 10% or less, preferably 5% or less, i.e., % low molecular weight species as measured, for example, by HP-SEC.Typically, antibody aggregation observed is less than 10%, preferably less than 5%, i.e., % high molecular weight species as measured, for example, by HP-SEC.

[0088] An antibody "retains physical stability" in a pharmaceutical formulation if it does not show a significant increase in aggregation, precipitation, and / or denaturation upon visual inspection of color and / or clarity, or as measured by UV light scattering, size exclusion chromatography (SEC), and dynamic light scattering. Changes in protein conformation can be assessed by fluorescence spectroscopy, which confirms the tertiary structure of a protein, and FTIR spectroscopy, which confirms the secondary structure of a protein.

[0089] An antibody "retains chemical stability" in a pharmaceutical formulation if it does not undergo significant chemical changes. Chemical stability can be assessed by detecting and quantifying chemically altered forms of the protein. Degradation processes that often alter the chemical structure of a protein include hydrolysis or clipping (assessed by methods such as size exclusion chromatography and SDS-PAGE), oxidation (assessed by methods such as MALDI / TOF / MS or peptide mapping in combination with mass spectrometry), deamidation (assessed by methods such as ion exchange chromatography, capillary isoelectric focusing, peptide mapping, isoaspartic acid measurement), and isomerization (assessed by measurement of isoaspartic acid content, peptide mapping, etc.).

[0090] An antibody is "biologically stable" in a pharmaceutical formulation if the biological activity of the antibody at a given time is within a given range of the biological activity exhibited at the time the pharmaceutical formulation was prepared. The biological activity of the antibody can be confirmed, for example, by antigen binding assays.

[0091] The term "isotonic" means that the formulation has essentially the same osmotic pressure as human blood. Isotonic formulations generally have an osmotic pressure of about 270-328 mOsm. Slightly hypotonic is 250-269 mOsm, and slightly hypertonic is 328-350 mOsm. Osmotic pressure can be measured, for example, using a vapor pressure or ice pressure osmometer.

[0092] As used herein, "co-formulated" or "co-formulation" or "coformulation" or "coformulated" refers to at least two different antibodies or antigen-binding fragments thereof that are formulated together and stored as a combination product in a single vial or container (e.g., an injection device), rather than being formulated and stored separately and then mixed and administered or administered separately. In one embodiment, the co-formulation comprises two different antibodies or antigen-binding fragments thereof.

[0093] Anti-ILT4 antibodies and antigen-binding fragments thereof The formulations disclosed herein can be used with any antibody or antigen-binding fragment thereof that binds to ILT4 (e.g., fully human antibodies). In one embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof is an antagonist. In another embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof is an anti-ILT4 antibody or antigen-binding fragment thereof described herein (e.g., 1E1, 2A6, 3G7, or 2C1).

[0094] In one aspect, the formulations can be used with the anti-ILT4 antibodies and antigen-binding fragments thereof described herein that have one or more of the following properties: * Binding human ILT4 to one or more amino acid residues of LYREKKSASW (SEQ ID NO: 59), TRIRPEL (SEQ ID NO: 60), NGQF (SEQ ID NO: 61) and / or HTGRYGCQ (SEQ ID NO: 62), and / or isolating LYREKKSASW (SEQ ID NO: 59), TRIRPEL (SEQ ID NO: 60), NGQF (SEQ ID NO: 61) and / or HTGRYGCQ (SEQ ID NO: 62) with deuterium (e.g., D 2 O) protect against exchange (e.g., as confirmed by hydrogen-deuterium exchange mass spectrometry); * Binds to human ILT4 via domain 1 (see Wilcox et al. BMC Structural Biology 2:6(2002)); * binds to the human ILT4 extracellular domain or the TM form of ILT4 expressed on the cell surface (e.g., pre-B cells, Chinese hamster ovary cells, U937 cells or Jurkat JE6 cells); * Calculated pI of about 7.29 (e.g., 7.29 or 7.30); *Experimentally determined pI approx. 7.2; * Characterized by a thermogram with a Tm onset >60°C, a Tm1 of about 65.2°C and a Tm2 of about 78.8°C; * Approximately 1.7×10 -8 K of M D binds to human ILT4 (e.g., as confirmed by surface plasmon resonance, e.g., by binding of anti-ILT4 to polyhistidine-tagged human ILT4); * Ka=5.5×10 5 M -1 s -1 (e.g., as confirmed by surface plasmon resonance, e.g., by binding of anti-ILT4 to polyhistidine-tagged human ILT4); * Kd=9×10 -3 s -1 (e.g., as confirmed by surface plasmon resonance, e.g., by binding of anti-ILT4 to polyhistidine-tagged human ILT4); * Binding of HLA-G (e.g., Fc-fused HLA-G) to human ILT4 (e.g., ILT4 on murine 3A9 T cells transfected with ILT4 and expressing ILT4) is measured, for example, with an IC of about 0.25 micrograms / mL (±0.06 micrograms / mL). 50 , blocking (e.g., as confirmed by surface plasmon resonance); * blocking the binding of HLA-A, HLA-B (e.g., fluorochrome-labeled dexamers of HLA-A, e.g., HLA*A2:01, or HLA-B, e.g., HLA*B7:02) and / or HLA-F (e.g., fluorochrome-labeled tetramers of HLA-F) to human ILT4 (e.g., ILT4 on murine 3A9 T cells transfected with ILT4 and expressing ILT4) (e.g., as determined by surface plasmon resonance); * blocking the binding of LT4 (e.g., ILT4 on murine 3A9 T cells transfected with ILT4 and expressing ILT4) to ANGPTL1, ANGPTL4 and / or ANGPTL7 (e.g., biotinylated ANGPTL proteins) (e.g., as determined by surface plasmon resonance); *Does not bind to ILT2, ILT3, ILT5, LILRB5, LILRA1, LILRA2, ILT7, ILT8 and / or ILT11; * In ILT4-transfected 3A9 cells, ILT4-mediated IL2 suppression was measured with an EC 50 And then reverse it; * For example, when mast cells express ILT4 and CD200RLa and are stimulated, for example, by antibody-mediated cross-linking of CD200RLa, rescues the ILT4:HLA-G-induced inhibition of mast cell degranulation (e.g., in the presence of plate-bound HLA-G tetramers); * Enhance lipopolysaccharide (LPS)-induced expression of proinflammatory myeloid cytokines (e.g., GM-CSF and / or TNFα) from peripheral blood mononuclear cells (PBMCs); * Enhance anti-CD3-induced expression of proinflammatory myeloid cytokines (e.g., GM-CSF and / or TNFα) from peripheral blood mononuclear cells (PBMCs); * inhibiting tumor growth in humans or, for example, in other mammals, for example, mice reconstituted with human hematopoietic stem cells harboring peripheral human CD45+ immune cells (e.g., immunodeficient NSG mice) (e.g., where the tumor is a human cutaneous melanoma tumor, for example, a human cutaneous melanoma tumor derived from cell line SKMEL5); * alleviating MDSC-mediated tumor tolerance in a tumor-bearing subject (e.g., a human subject); *Does not bind to cynomolgus ILT4 and / or mouse pirB; * staining CD14+ human monocytes and / or CD11B+ human granulocytes; and / or * Binds to one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or all 10) of the human ILT4 haplotypes.

[0095] Antibody 1E1 (Q1E) heavy chain (IgG4) Heavy chain EVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLPTRWVTTRYFDLWGRGTLVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:1; variable domains underlined; CDRs double underlined).

[0096] Heavy Chain Variable Domain EVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLPTRWVTTRYFDLWGRGTLVTVSS (SEQ ID NO:63).

[0097] Antibody 1E1 (Q1E, S54A) heavy chain (IgG4) Heavy chain EVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHAGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLPTRWVTTRYFDLWGRGTLVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:2; variable domains underlined; CDRs double underlined).

[0098] Heavy Chain Variable Domain EVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHAGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLPTRWVTTRYFDLWGRGTLVTVSS (SEQ ID NO:57).

[0099] Antibody 1E1 heavy chain (IgG1) Heavy chain QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLPTRWVTTRYFDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:44; variable domain underlined; CDRs double underlined).

[0100] Heavy Chain Variable Domain QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLPTRWVTTRYFDLWGRGTLVTVSS (sequence number: 69).

[0101] 1E1 Heavy Chain CDRs CDR-H1: GYYWS (SEQ ID NO: 16) CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17) (e.g., EINHSGSTNYNPSLKS (SEQ ID NO: 47) or EINHAGSTNYNPSLKS (SEQ ID NO: 48)) CDR-H3: LPTRWVTTRYFDL (sequence number: 18).

[0102] Antibody 1E1 (Q1E) light chain (lambda) Light chain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:3; variable domains underlined; CDRs double underlined).

[0103] Light Chain Variable Domain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL (SEQ ID NO: 70).

[0104] Antibody 1E1 (Q1E, S54A) light chain (lambda) Light chain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNANRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:4; variable domains underlined; CDRs double underlined).

[0105] Light Chain Variable Domain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNANRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL (Sequence number: 71).

[0106] Antibody 1E1 (Q1E, N53Q) light chain (lambda) Light chain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGQSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:5; variable domains underlined; CDRs double underlined).

[0107] Light Chain Variable Domain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGQSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL (Sequence number: 72).

[0108] Antibody 1E1 (Q1E, N53E) light chain (lambda) Light chain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGESNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:6; variable domains underlined; CDRs double underlined).

[0109] Light Chain Variable Domain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGESNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL (Sequence number: 73).

[0110] Antibody 1E1 (Q1E, N53D) light chain (lambda) Light chain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGDSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:7; variable domain underlined; CDRs double underlined).

[0111] Light Chain Variable Domain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGDSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL (Sequence number: 58).

[0112] Antibody 1E1 light chain (lambda) Light chain QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:45; variable domain underlined; CDRs double underlined).

[0113] Light Chain Variable Domain QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL (Sequence number: 77).

[0114] 1E1 Light Chain CDRs CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19) CDR-L2:GX 1 X 2 NRPS; where X 1 is N, Q, E or D, and X 2 is S or A (SEQ ID NO: 20) (e.g., GNSNRPS (SEQ ID NO: 49), GQSNRPS (SEQ ID NO: 50), GESNRPS (SEQ ID NO: 51), GDSNRPS (SEQ ID NO: 52), GNANRPS (SEQ ID NO: 53), GQANRPS (SEQ ID NO: 54), GEANRPS (SEQ ID NO: 55), or GDANRPS (SEQ ID NO: 56)) CDR-L3: QSFDNSLSAYV (sequence number: 21).

[0115] 1E1 heavy and light chain CDRs or 1E1 V H and V L Alternatively, antibodies and antigen-binding fragments thereof comprising 1E1 heavy and light chains (or variants thereof, such as those described herein) may be referred to as "1E1."

[0116] Antibody 2A6(Q1E) Heavy Chain (IgG4) Heavy chain EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARYFDSSGWYKGGAFDIWGQGTMVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:8; variable domains underlined; CDRs double underlined).

[0117] Heavy Chain Variable Domain EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARYFDSSGWYKGGAFDIWGQGTMVTVSS (Sequence number: 64).

[0118] Antibody 2A6 (Q1E, S102A, M119L) heavy chain (IgG4) Heavy chain EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARYFDASGWYKGGAFDIWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:9; variable domains underlined; CDRs double underlined).

[0119] Heavy Chain Variable Domain EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARYFDASGWYKGGAFDIWGQGTLVTVSS (Sequence number: 65).

[0120] Antibody 2A6 (Q1E, D101S, M119L) heavy chain (IgG4) Heavy chain EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARYFSSSGWYKGGAFDIWGQGTLVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:10; variable domain underlined; CDRs double underlined).

[0121] Heavy Chain Variable Domain EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARYFSSSGWYKGGAFDIWGQGTLVTVSS (Sequence number: 66).

[0122] The formulations disclosed herein can be used with antibodies and antigen-binding fragments thereof comprising SEQ ID NO:8, 9, 10, 64, 65 or 66, where residue 1 of SEQ ID NO:8, 9, 10, 64, 65 or 66 is Q instead of E.

[0123] 2A6 Heavy Chain CDRs CDR-H1: SYAIS (SEQ ID NO: 22) CDR-H2: GIIPIFGTANYAQKFQG (SEQ ID NO: 23) CDR-H3:YFX 1 X 2 SGWYKGGAFDI;where X 1 is D or S, and X 2 is S or A (SEQ ID NO: 24) (e.g., YFDSSGWYKGGAFDI (SEQ ID NO: 91), YFSSSGWYKGGAFDI (SEQ ID NO: 92), YFDASGWYKGGAFDI (SEQ ID NO: 93), or YFSASGWYKGGAFDI (SEQ ID NO: 94)).

[0124] Antibody 2A6 light chain (lambda) Light chain QSVLTQPSSLSASPGASASLTCTLRSGINVDTYRIHWYQQKPGSPPQYLLRYKSDSDKHQGSGVPSRFSGSKDPSANAGILLISGLQSEDEADYYCAIWYSSTWVFGGGTQLTVL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:11; variable domain underlined; CDRs double underlined).

[0125] Light Chain Variable Domain QSVLTQPSSLSASPGASASLTCTLRSGINVDTYRIHWYQQKPGSPPQYLLRYKSDSDKHQGSGVPSRFSGSKDPSANAGILLISGLQSEDEADYYCAIWYSSTWVFGGGTQLTVL (Sequence number: 74).

[0126] The formulations disclosed herein can be used with antibodies and antigen-binding fragments thereof comprising SEQ ID NO:11 or 74, where residue 1 of SEQ ID NO:11 or 74 is E instead of Q.

[0127] 2A6 light chain CDRs CDR-L1: TLRSGINVDTYRIH (SEQ ID NO: 25) CDR-L2: YKSDSDKHQGS (SEQ ID NO: 26) CDR-L3: AIWYSSTWV (sequence number: 27).

[0128] 2A6 heavy and light chain CDRs or 2A6 V H and V L Alternatively, antibodies and antigen-binding fragments thereof comprising the 2A6 heavy and light chains (or variants thereof, such as those described herein) may be referred to as "2A6."

[0129] Antibody 3G7(Q1E) heavy chain (IgG4) Heavy chain EVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVGEWIQLWSPFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:12; variable domain underlined; CDRs double underlined).

[0130] Heavy Chain Variable Domain EVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVGEWIQLWSPFDYWGQGTLVTVSS (Sequence number: 67).

[0131] The formulations disclosed herein can be used with antibodies and antigen-binding fragments thereof comprising SEQ ID NO: 12 or 67, where residue 1 of SEQ ID NO: 12 or 67 is Q instead of E.

[0132] 3G7 Heavy Chain CDR CDR-H1: SYAMH (SEQ ID NO: 28) CDR-H2: VISYDGSNKYYADSVKG (SEQ ID NO: 29) CDR-H3: VGEWIQLWSPFDY (sequence number: 30).

[0133] Antibody 3G7 light chain (kappa) Light chain DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKFLIYAASSLQSGVPSKFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSYPPTFGGGTKVEIKRtVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:13; variable domain underlined; CDRs double underlined).

[0134] Light Chain Variable Domain DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKFLIYAASSLQSGVPSKFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSYPPTFGGGTKVEIK (Sequence number: 75).

[0135] 3G7 light chain CDR CDR-L1: RASQGISSWLA (SEQ ID NO: 31) CDR-L2: AASSLQS (SEQ ID NO: 32) CDR-L3: QQYNSYPPT (sequence number: 33).

[0136] 3G7 heavy and light chain CDRs or 3G7 V H and V L Alternatively, antibodies and antigen-binding fragments thereof comprising the 3G7 heavy and light chains (or variants thereof, such as those described herein) may be referred to as "3G7."

[0137] Antibody 2C1(Q1E) Heavy Chain (IgG4) Heavy chain EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMGGFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCARAGPLYTIFGVVIIPDNWFDPWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:14; variable domain underlined; CDRs double underlined).

[0138] Heavy Chain Variable Domain EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMGGFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCARAGPLYTIFGVVIIPDNWFDPWGQGTLVTVSS (Sequence number: 68).

[0139] The formulations disclosed herein can be used with antibodies and antigen-binding fragments thereof comprising SEQ ID NO: 14 or 68, where residue 1 of SEQ ID NO: 14 or 68 is Q instead of E.

[0140] 2C1 Heavy Chain CDR CDR-H1: ELSMH (SEQ ID NO: 34) CDR-H2: GFDPEDGETIYAQKFQG (SEQ ID NO: 35) CDR-H3: AGPLYTIFGVVIIPDNWFDP (sequence number: 36).

[0141] Antibody 2C1 light chain (Q1E) (lambda) Light chain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSGVVFGGGTQLIILGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:15; variable domains underlined; CDRs double underlined).

[0142] Light Chain Variable Domain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSGVVFGGGTQLIIL (Sequence number: 76).

[0143] The formulations disclosed herein can be used with antibodies and antigen-binding fragments thereof comprising SEQ ID NO: 15 or 76, where residue 1 of SEQ ID NO: 15 or 76 is Q instead of E.

[0144] 2C1 light chain CDRs CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 37) CDR-L2: GNSNRPS (SEQ ID NO: 38) CDR-L3: QSYDSSLSGSGVV (sequence number: 39).

[0145] 2C1 heavy and light chain CDRs or 2C1 V H and V L Alternatively, antibodies and antigen-binding fragments thereof comprising 2C1 heavy and light chains (or variants thereof, such as those described herein) may be referred to as "2C1."

[0146] In various embodiments of the antibody or antigen-binding fragment thereof, the C-terminal lysine of the heavy chain immunoglobulin is absent.

[0147] Thus, in some embodiments, the antibody or antigen-binding fragment thereof comprises a light chain immunoglobulin, a heavy chain immunoglobulin, or both a light chain immunoglobulin and a heavy chain immunoglobulin, where the light chain immunoglobulin comprises the amino acid sequence set forth in SEQ ID NO:3, 4, 5, 6, 7, 11, 13, 15 or 45; and / or the heavy chain immunoglobulin comprises the amino acid sequence set forth in SEQ ID NO:1, 2, 8, 9, 10, 12, 14, 44, 79, 80, 81, 82, 83, 84, 85 or 86.

[0148] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:1 or 79; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:3.

[0149] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:2 or 80; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:4.

[0150] In another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:2 or 80; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:5.

[0151] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:2 or 80; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:6.

[0152] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:2 or 80; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:7.

[0153] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:2 or 80; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:3.

[0154] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:8 or 82; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:11.

[0155] In another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:9 or 83; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:11.

[0156] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:10 or 84; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:11.

[0157] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 12 or 85; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 13.

[0158] In a further embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:14 or 86; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:15.

[0159] In certain further embodiments, the antibody or antigen-binding fragment thereof comprises a light chain immunoglobulin, a heavy chain immunoglobulin, or both a light chain immunoglobulin and a heavy chain, wherein the light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 70, 71, 72, 73, 58, 74, 75, 76, or 77, and / or the heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 63, 57, 64, 65, 66, 67, 68, or 69.

[0160] In certain embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:63; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:70.

[0161] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:71.

[0162] In another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:72.

[0163] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:73.

[0164] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:58.

[0165] In certain embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:70.

[0166] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:64; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:74.

[0167] In another embodiment, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:65; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:74.

[0168] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:66; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:74.

[0169] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:67; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:75.

[0170] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:68; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:76.

[0171] In a further embodiment, the antibody or antigen-binding fragment thereof that binds to ILT4 has an immunoglobulin light chain variable (VV) domain comprising CDR-L1, CDR-L2 and CDR-L3 of 1E1 (e.g., SEQ ID NOs: 19-21). L ) domain; and an immunoglobulin heavy chain variable (V) domain comprising CDR-H1, CDR-H2 and CDR-H3 of 1E1 (e.g., SEQ ID NO:16-18). H ) domain;

[0172] In a further embodiment, the antibody or antigen-binding fragment thereof that binds ILT4 has an immunoglobulin light chain variable (VV) domain comprising CDR-L1, CDR-L2 and CDR-L3 of 2A6 (e.g., SEQ ID NOs:25-27). L ) domain; and an immunoglobulin heavy chain variable (V) domain comprising the CDR-H1, CDR-H2 and CDR-H3 of 2A6 (e.g., SEQ ID NO:22-24). H ) domain;

[0173] In a further embodiment, the antibody or antigen-binding fragment thereof that binds to ILT4 has an immunoglobulin light chain variable (VV) domain comprising CDR-L1, CDR-L2 and CDR-L3 of 3G7 (e.g., SEQ ID NOs: 31-33). L ) domain; and an immunoglobulin heavy chain variable (V) domain comprising the CDR-H1, CDR-H2 and CDR-H3 of 3G7 (e.g., SEQ ID NO:28-30). H ) domain;

[0174] In a further embodiment, the antibody or antigen-binding fragment thereof that binds ILT4 has an immunoglobulin light chain variable (VV) domain comprising CDR-L1, CDR-L2 and CDR-L3 of 2C1 (e.g., SEQ ID NOs: 37-39). L ) domain; and an immunoglobulin heavy chain variable (V) domain comprising CDR-H1, CDR-H2 and CDR-H3 of 2C1 (e.g., SEQ ID NOs: 34-36). H ) domain;

[0175] In one embodiment, the antibody or antigen-binding fragment comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GNSNRPS (SEQ ID NO: 49), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0176] In another embodiment, the antibody or antigen-binding fragment comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GQSNRPS (SEQ ID NO: 50), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0177] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GESNRPS (SEQ ID NO: 51), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0178] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0179] In one embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). Hdomain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GNANRPS (SEQ ID NO: 53), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0180] In another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GQANRPS (SEQ ID NO: 54), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0181] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GEANRPS (SEQ ID NO: 55), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0182] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDANRPS (SEQ ID NO: 56), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0183] In one embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GNSNRPS (SEQ ID NO: 49), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0184] In another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GQSNRPS (SEQ ID NO: 50), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0185] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GESNRPS (SEQ ID NO: 51), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0186] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0187] In one embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GNANRPS (SEQ ID NO: 53), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0188] In another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GQANRPS (SEQ ID NO: 54), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0189] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). Hdomain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GEANRPS (SEQ ID NO: 55), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0190] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDANRPS (SEQ ID NO: 56), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0191] The formulations disclosed herein bind to ILT4 and have the V L Domains (e.g., SEQ ID NOs: 70, 71, 72, 73, 58, or 77) and / or V of antibody 1E1 H The antibody or antigen-binding fragment thereof may be used in combination with an antibody comprising the domain (e.g., SEQ ID NO: 63, 57, or 69).

[0192] The formulations disclosed herein bind to ILT4 and are L Domain (e.g., SEQ ID NO:74) and / or V of antibody 2A6 H It may be used in conjunction with an antibody or antigen-binding fragment thereof comprising the domain (e.g., SEQ ID NO: 64, 65 or 66).

[0193] The formulations disclosed herein bind to ILT4 and are L Domains (e.g., SEQ ID NO:75) and / or V of antibody 3G7 H The antibody or antigen-binding fragment thereof may be used in combination with an antibody comprising the domain (eg, SEQ ID NO:67).

[0194] The formulations disclosed herein bind to ILT4 and have the V L Domain (e.g., SEQ ID NO:76) and / or V of antibody 2C1 H It may be used in conjunction with an antibody or antigen-binding fragment thereof that contains the domain (eg, SEQ ID NO:68).

[0195] The formulations disclosed herein may be used with antibodies or antigen-binding fragments thereof that bind ILT4 and include the light chain immunoglobulin of antibody 1E1 (e.g., SEQ ID NO:3, 4, 5, 6, 7 or 45) and / or the heavy chain immunoglobulin of antibody 1E1 (e.g., SEQ ID NO:1, 2, 44, 79, 80 or 81).

[0196] The formulations disclosed herein may be used with antibodies or antigen-binding fragments thereof that bind ILT4 and include the light chain immunoglobulin of antibody 2A6 (e.g., SEQ ID NO:11) and / or the heavy chain immunoglobulin of antibody 2A6 (e.g., SEQ ID NOs:8, 9, 10, 82, 83 or 84).

[0197] The formulations disclosed herein may be used with an antibody or antigen-binding fragment thereof that binds ILT4 and includes the light chain immunoglobulin of antibody 3G7 (e.g., SEQ ID NO:13) and / or the heavy chain immunoglobulin of antibody 3G7 (e.g., SEQ ID NO:12 or 85).

[0198] The formulations disclosed herein may be used with an antibody or antigen-binding fragment thereof that binds ILT4 and comprises the light chain immunoglobulin of antibody 2C1 (e.g., SEQ ID NO:15) and / or the heavy chain immunoglobulin of antibody 2C1 (e.g., SEQ ID NO:14 or 86).

[0199] The formulations disclosed herein can be used with antibodies that consist of two heavy chains and two light chains, where each light chain is a V of antibody 1E1, 2A6, 3G7, or 2C1. Lor light chain immunoglobulin, each heavy chain comprising the V of antibody 1E1, 2A6, 3G7 or 2C1 H or heavy chain immunoglobulins.

[0200] In one embodiment, the antibody consists of two heavy chains and two light chains, where each light chain comprises the amino acid sequence set forth in SEQ ID NO:58 and each heavy chain comprises the amino acid sequence set forth in SEQ ID NO:57.

[0201] In another embodiment, the antibody consists of two heavy chains and two light chains, wherein each light chain comprises the amino acid sequence set forth in SEQ ID NO:58, each heavy chain comprises the amino acid sequence set forth in SEQ ID NO:57, and the light chains further comprise the amino acid sequence set forth in SEQ ID NO:90.

[0202] In yet another embodiment, the antibody consists of two heavy chains and two light chains, wherein each light chain comprises the amino acid sequence set forth in SEQ ID NO:58 and each heavy chain comprises the amino acid sequence set forth in SEQ ID NO:57, wherein the heavy chains further comprise the amino acid sequence set forth in SEQ ID NO:89.

[0203] In yet another embodiment, the antibody consists of two heavy chains and two light chains, wherein each light chain comprises the amino acid sequence set forth in SEQ ID NO:58 and each heavy chain comprises the amino acid sequence set forth in SEQ ID NO:57, wherein the light chain further comprises the amino acid sequence set forth in SEQ ID NO:90 and the heavy chain further comprises the amino acid sequence set forth in SEQ ID NO:89.

[0204] In one embodiment, the antibody consists of two heavy chains and two light chains, where each light chain comprises the amino acid sequence set forth in SEQ ID NO:7 and each heavy chain comprises the amino acid sequence set forth in SEQ ID NO:2.

[0205] In another embodiment, the antibody consists of two heavy chains and two light chains, wherein each light chain consists of the amino acid sequence set forth in SEQ ID NO:7 and each heavy chain consists of the amino acid sequence set forth in SEQ ID NO:2.

[0206] In one embodiment, the antibody or antigen-binding fragment thereof has the V L(with or without a signal sequence), e.g., one, two, three, four, five, six, seven, eight, nine, ten or more conservative or non-conservative amino acid substitutions, such as V in any of SEQ ID NOs: 58 or 70-77 L and / or V H 57 or 63-69 (with or without a signal sequence), e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more conservative or non-conservative amino acid substitutions and still bind to ILT4 H ;includes.

[0207] The formulations disclosed herein can be used with polypeptides that include an amino acid sequence disclosed herein (e.g., SEQ ID NOs:1-39, 44, 45, 47-58, 63-77 or 79-86), as well as with polypeptides that include said amino acid sequences having up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 20 or more conservative or non-conservative amino acid substitutions therein.

[0208] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a light chain immunoglobulin, a heavy chain immunoglobulin, or both a light chain immunoglobulin and a heavy chain immunoglobulin, wherein the light chain immunoglobulin has at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:3, 4, 5, 6, 7, 11, 13, 15 or 45, and / or the heavy chain immunoglobulin has at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:1, 2, 8, 9, 10, 12, 14, 44, 79, 80, 81, 82, 83, 84, 85 or 86.

[0209] In some embodiments, an antibody or antigen-binding fragment thereof comprises a light chain immunoglobulin, a heavy chain immunoglobulin, or both a light chain immunoglobulin and a heavy chain immunoglobulin, wherein the light chain immunoglobulin comprises a light chain variable domain having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 70, 71, 72, 73, 58, 74, 75, 76 or 77, and / or the heavy chain immunoglobulin comprises a heavy chain variable domain having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 63, 57, 64, 65, 66, 67, 68 or 69.

[0210] In one embodiment, the immunoglobulin heavy chain of the anti-ILT4 antibody or antigen-binding fragment of the invention is operably linked to a signal sequence (e.g., a signal sequence comprising the amino acid sequence MEWSWVFLFFLSVTTGVHS (SEQ ID NO: 41)) and / or the immunoglobulin light chain of the anti-ILT4 antibody or antigen-binding fragment of the invention is operably linked to a signal sequence (e.g., a signal sequence comprising the amino acid sequence MSVPTQVLGLLLLWLTDARC (SEQ ID NO: 42)).

[0211] In one embodiment, the N-terminal glutamine (Q) of the immunoglobulin chains (e.g., heavy and / or light chains) described herein is replaced with pyroglutamic acid. In one embodiment, the N-terminal Q of the heavy chain immunoglobulin is replaced with pyroglutamic acid. In another embodiment, the N-terminal Q of the light chain immunoglobulin is replaced with pyroglutamic acid. In yet another embodiment, the N-terminal Q of the heavy chain immunoglobulin and the N-terminal Q of the heavy chain immunoglobulin are replaced with pyroglutamic acid.

[0212] The formulations disclosed herein can also be used with any of the anti-ILT4 antibodies or antigen-binding fragments thereof disclosed herein (e.g., 1E1, 2A6, 3G7, or 2C1) and an antibody or antigen-binding fragment that binds to the same epitope of ILT4 (e.g., human ILT4). In one embodiment, the epitope is LYREKKSASW (SEQ ID NO:59). In another embodiment, the epitope is TRIRPEL (SEQ ID NO:60). In yet another embodiment, the epitope is NGQF (SEQ ID NO:61). In yet another embodiment, the epitope is HTGRYGCQ (SEQ ID NO:62). In certain embodiments, the antibody or antigen-binding fragment thereof binds to the same epitope of human ILT4 as an antibody or antigen-binding fragment thereof comprising the heavy chain and light chain amino acid sequences set forth in SEQ ID NOs:1 and 3; 2 and 4; 2 and 5; 2 and 6; 2 and 7; 2 and 3; 8 and 11; 9 and 11; 10 and 11; 12 and 13; 14 and 15; 79 and 3; 80 and 4; 80 and 5; 80 and 6; 80 and 7; 80 and 3; 82 and 11; 83 and 11; 84 and 11; 85 and 13; and 86 and 15; respectively. In some embodiments, the antibody or antigen-binding fragment thereof binds to the same epitope of human ILT4 as an antibody or antigen-binding fragment thereof comprising the heavy chain variable domain amino acid sequences and the light chain variable domain amino acid sequences set forth in SEQ ID NOs: 63 and 70; 57 and 71; 57 and 72; 57 and 73; 57 and 58; 57 and 70; 64 and 74; 65 and 74; 66 and 74; 67 and 75; 68 and 76; respectively.

[0213] The formulations disclosed herein can be used with antibodies or antigen-binding fragments that cross-block the binding of any of the anti-ILT4 antibodies or antigen-binding fragments thereof disclosed herein (e.g., 1E1, 2A6, 3G7 or 2C1) to ILT4 (e.g., human ILT4) or with antibodies and antigen-binding fragments that compete with the binding of any of the anti-ILT4 antibodies or antigen-binding fragments thereof disclosed herein (e.g., 1E1, 2A6, 3G7 or 2C1) to ILT4 (e.g., human ILT4). The cross-blocking antibodies and antigen-binding fragments thereof discussed herein can be confirmed based on their ability to block any of the antibodies or fragments specifically described herein from binding to ILT4 in binding assays (e.g., biolayer interferometry (BLI; e.g., FORTEBIO OCTET binding assay; Pall ForteBio Corp; Menlo Park, CA), surface plasmon resonance (SPR), BIACore, ELISA, flow cytometry). For example, in an embodiment of the invention, when using BLI, the tip of a fiber optic probe is coated with a ligand (e.g., ILT4) and functions as a biosensor, where binding of an anti-ILT4 antibody or antigen binding fragment to ILT4 changes the interference pattern of white light reflected from the ILT4-bound probe layer and the internal reference layer. This shift is indicative of ILT4 / anti-ILT4 binding. In an embodiment of the invention, the ILT4-coated tip is immersed in a solution of analyte containing antibodies or antigen binding fragments, for example in the wells of either a 96-well plate or a 384-well plate. In an embodiment of the invention, the plate is shaken during reading to create orbital flow. To read the assay, white light is directed down the length of the fiber. As described above, interference of the light reflected from the reference layer with the immobilization surface containing ILT4 on the tip produces a characteristic pattern of light reflected back up the fiber. When molecules bind to the immobilized sensor surface, the pattern changes in proportion to the extent of binding.For example, an assay can be used in which ILT4 (e.g., human ILT4) protein is immobilized on a BLI probe or plate, a reference anti-ILT4 antibody or fragment is bound to ILT4 (e.g., at saturating concentration), and a test anti-ILT4 antibody or fragment is added. The ability of the test antibody to compete with the reference antibody for ILT4 binding is then measured. In a BLI format, the light interference of the ILT4 complex is monitored to confirm whether the test antibody effectively competes with the reference antibody. For example, nanometers of the wavelength shift of light over time are monitored, where the shift indicates additional binding and lack of cross-blocking of the test antibody. In an embodiment of the invention, in a BLI format, if no additional binding of the test antibody is observed, cross-blocking between the antibodies is deemed to have occurred qualitatively. In an embodiment of the invention, as a control, cross-blocking of the reference antibody with itself is confirmed; where if the reference antibody is able to cross-block itself from ILT4 binding, the assay is confirmed to be working correctly. The ability of the test antibody to inhibit the binding of anti-ILT4 antibody or fragment 1E1, 2A6, 3G7 or 2C1 to ILT4 (e.g. human ILT4) indicates that the test antibody is capable of cross-blocking the antibody or fragment for binding to ILT4 (e.g. human ILT4) and therefore, in some cases, may bind to the same epitope on ILT4 (e.g. human ILT4) as 1E1, 2A6, 3G7 and / or 2C1. As mentioned above, antibodies and fragments that bind to the same epitope as any of the anti-ILT4 antibodies or fragments of the invention also form part of the invention. In one embodiment of the invention, BLI is performed in a sandwich format, where a reference anti-ILT4 antibody or antigen-binding fragment is immobilized on a probe and then binds to ILT4. The test anti-ILT4 antibody or antigen-binding fragment is then tested for its ability to block the binding of the reference antibody or fragment.

[0214] In certain embodiments, the antibody or antigen-binding fragment thereof competes for binding to human ILT4 with an antibody or fragment comprising the heavy and light chain amino acid sequences set forth in SEQ ID NOs:1 and 3; 2 and 4; 2 and 5; 2 and 6; 2 and 7; 2 and 3; 8 and 11; 9 and 11; 10 and 11; 12 and 13; 14 and 15; 79 and 3; 80 and 4; 80 and 5; 80 and 6; 80 and 7; 80 and 3; 82 and 11; 83 and 11; 84 and 11; 85 and 13; and 86 and 15; respectively. In some embodiments, the antibody or antigen-binding fragment thereof competes for binding to human ILT4 with an antibody or fragment comprising the heavy chain variable domain amino acid sequences and the light chain variable domain amino acid sequences set forth in SEQ ID NOs: 63 and 70; 57 and 71; 57 and 72; 57 and 73; 57 and 58; 57 and 70; 64 and 74; 65 and 74; 66 and 74; 67 and 75; 68 and 76; respectively.

[0215] The formulations disclosed herein can be used with anti-ILT4 antibodies and antigen-binding fragments thereof that include N-linked glycans typically added to immunoglobulins produced in Chinese hamster ovary cells (CHO N-linked glycans) or with anti-ILT4 antibodies and antigen-binding fragments thereof that include N-linked glycans typically added to immunoglobulins produced in artificial yeast cells (e.g., Pichia pastoris) (artificial yeast N-linked glycans). For example, in embodiments, the anti-ILT4 antibodies and antigen-binding fragments thereof include one or more of "artificial yeast N-linked glycans" or "CHO ​​N-linked glycans" (e.g., G0 and / or G0-F and / or G1 and / or G1-F and / or G2-F and / or Man5). In embodiments, the anti-ILT4 antibodies and antigen-binding fragments thereof include artificial yeast N-linked glycans (i.e., G0 and / or G1 and / or G2, optionally further including Man5). In embodiments, the anti-ILT4 antibodies and antigen-binding fragments thereof comprise CHO N-linked glycans (i.e., G0-F, G1-F and G2-F, optionally further including G0 and / or G1 and / or G2 and / or Man5). In embodiments, about 80% to about 95% (e.g., about 80-90%, about 85%, about 90% or about 95%) of the total N-linked glycans on the anti-ILT4 antibodies and antigen-binding fragments thereof are artificial yeast N-linked glycans or CHO N-linked glycans. See, Nett et al. Yeast. 28(3): 237-252 (2011); Hamilton et al. Science. 313(5792): 1441-1443 (2006); Hamilton et al. Curr Opin Biotechnol. 18(5): 387-392 (2007). For example, in embodiments, the artificial yeast cell is GFI5.0 or YGLY8316, or a strain described in U.S. Patent No. 7,795,002 or Zha et al., Methods Mol Biol. 988:31-43 (2013). See also International Patent Application Publication No. WO2013 / 066765.

[0216] Anti-ILT4 antibodies and antigen-binding fragments thereof (e.g., 1E1, 2A6, 3G7, and / or 2C1) that can be used with the various formulations disclosed herein can also be engineered to include modifications within the Fc region, typically to alter one or more functional properties of the antibody (e.g., serum half-life, complement fixation, Fc receptor binding, and / or effector function (e.g., antigen-dependent cellular cytotoxicity), etc.). Additionally, antibodies and antigen-binding fragments thereof (e.g., 1E1, 2A6, 3G7, and / or 2C1) that can be used with the various formulations disclosed herein can be chemically modified (e.g., one or more chemical moieties can be attached to the antibody, such as a labeled antibody or antibody-drug conjugate) or modified to alter glycosylation, also to alter one or more functional properties of the antibody.

[0217] PD-1 antibodies and their antigen-binding fragments In one aspect, the invention provides stable biological formulations comprising an anti-ILT4 antibody or antigen-binding fragment thereof co-formulated with an anti-human PD-1 antibody or antigen-binding fragment thereof that specifically binds to human PD-1 (e.g., a human or humanized anti-PD-1 antibody) as the active pharmaceutical ingredient (PD-1 API), and methods of using the formulations of the invention. Any anti-PD-1 antibody or antigen-binding fragment thereof can be used in the co-formulations and methods of the invention. In certain embodiments, the PD-1 API is an anti-PD-1 antibody selected from pembrolizumab and nivolumab. In certain embodiments, the anti-PD-1 antibody is pembrolizumab. In alternative embodiments, the anti-PD-1 antibody is nivolumab. Table 3 provides the amino acid sequences of representative anti-human PD-1 antibodies pembrolizumab and nivolumab. Alternative PD-1 antibodies and antigen-binding fragments useful in the co-formulations and methods of the invention are shown in Table 4.

[0218] In some embodiments, an anti-human PD-1 antibody, or antigen-binding fragment thereof, for use in the co-formulations of the invention comprises the three light chain CDRs, CDRL1, CDRL2, and CDRL3, and / or the three heavy chain CDRs, CDRH1, CDRH2, and CDRH3.

[0219] In one embodiment of the invention, CDRL1 is SEQ ID NO:95 or a variant of SEQ ID NO:95, CDRL2 is SEQ ID NO:96 or a variant of SEQ ID NO:96, and CDRL3 is SEQ ID NO:97 or a variant of SEQ ID NO:97.

[0220] In one embodiment, CDRH1 is SEQ ID NO:100 or a variant of SEQ ID NO:100, CDRH2 is SEQ ID NO:101 or a variant of SEQ ID NO:101, and CDRH3 is SEQ ID NO:102 or a variant of SEQ ID NO:102.

[0221] In one embodiment, the three light chain CDRs are SEQ ID NO:95, SEQ ID NO:96 and SEQ ID NO:97, and the three heavy chain CDRs are SEQ ID NO:100, SEQ ID NO:101 and SEQ ID NO:102.

[0222] In an alternative embodiment of the invention, CDRL1 is SEQ ID NO:105 or a variant of SEQ ID NO:105, CDRL2 is SEQ ID NO:106 or a variant of SEQ ID NO:106, and CDRL3 is SEQ ID NO:107 or a variant of SEQ ID NO:107.

[0223] In one embodiment, CDRH1 is SEQ ID NO:110 or a variant of SEQ ID NO:110, CDRH2 is SEQ ID NO:111 or a variant of SEQ ID NO:111, and CDRH3 is SEQ ID NO:11128 or a variant of SEQ ID NO:112.

[0224] In one embodiment, the three light chain CDRs are SEQ ID NO:95, SEQ ID NO:96 and SEQ ID NO:97, and the three heavy chain CDRs are SEQ ID NO:100, SEQ ID NO:101 and SEQ ID NO:102.

[0225] In an alternative embodiment, the three light chain CDRs are SEQ ID NO:105, SEQ ID NO:106 and SEQ ID NO:107, and the three heavy chain CDRs are SEQ ID NO:110, SEQ ID NO:111 and SEQ ID NO:112.

[0226] In a further embodiment of the invention, CDRL1 is SEQ ID NO:115 or a variant of SEQ ID NO:115, CDRL2 is SEQ ID NO:116 or a variant of SEQ ID NO:116, and CDRL3 is SEQ ID NO:117 or a variant of SEQ ID NO:117.

[0227] In yet another embodiment, CDRH1 is SEQ ID NO:118 or a variant of SEQ ID NO:118, CDRH2 is SEQ ID NO:119 or a variant of SEQ ID NO:119, and CDRH3 is SEQ ID NO:120 or a variant of SEQ ID NO:120.

[0228] In another embodiment, the three light chain CDRs are SEQ ID NO:115, SEQ ID NO:116 and SEQ ID NO:117, and the three heavy chain CDRs are SEQ ID NO:118, SEQ ID NO:119 and SEQ ID NO:120.

[0229] Some anti-human PD-1 antibodies and antigen-binding fragments of the invention comprise a light chain variable region and a heavy chain variable region. In some embodiments, the light chain variable region comprises SEQ ID NO:98 or a variant of SEQ ID NO:98, and the heavy chain variable region comprises SEQ ID NO:103 or a variant of SEQ ID NO:103. In further embodiments, the light chain variable region comprises SEQ ID NO:108 or a variant of SEQ ID NO:108, and the heavy chain variable region comprises SEQ ID NO:113 or a variant of SEQ ID NO:113. In further embodiments, the heavy chain variable region comprises SEQ ID NO:121 or a variant of SEQ ID NO:121, and the light chain variable region comprises SEQ ID NO:122 or a variant of SEQ ID NO:122, SEQ ID NO:123 or a variant of SEQ ID NO:123, or a variant of SEQ ID NO:124 or a variant of SEQ ID NO:124. In such embodiments, the variant light or heavy chain variable region sequences are identical to the reference sequences except for having one, two, three, four, or five amino acid substitutions. In some embodiments, the substitutions are in the framework regions (i.e., outside the CDRs), hi some embodiments, one, two, three, four or five of the amino acid substitutions are conservative substitutions.

[0230] In one embodiment of a co-formulation of the invention, the anti-human PD-1 antibody or antigen-binding fragment comprises a light chain variable region that comprises or consists of SEQ ID NO:98, and a heavy chain variable region that comprises or consists of SEQ ID NO:103. In a further embodiment, the anti-human PD-1 antibody or antigen-binding fragment comprises a light chain variable region that comprises or consists of SEQ ID NO:108, and a heavy chain variable region that comprises or consists of SEQ ID NO:113. In one embodiment of a formulation of the invention, the anti-human PD-1 antibody or antigen-binding fragment comprises a light chain variable region that comprises or consists of SEQ ID NO:122, and a heavy chain variable region that comprises or consists of SEQ ID NO:121. In a further embodiment, the anti-human PD-1 antibody or antigen-binding fragment comprises a light chain variable region that comprises or consists of SEQ ID NO:123, and a heavy chain variable region that comprises or consists of SEQ ID NO:121. In another embodiment, the antibody or antigen-binding fragment comprises a light chain variable region that comprises or consists of SEQ ID NO:124 and a heavy chain variable region that comprises or consists of SEQ ID NO:121.

[0231] In another embodiment, the co-formulation of the invention comprises an anti-human PD-1 antibody or antigen binding protein having a VL domain and / or a VH domain that exhibits at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, 80%, 75% or 50% sequence homology to one of the VL domains or VH domains set forth above and exhibits specific binding to PD-1. In another embodiment, the anti-human PD-1 antibody or antigen binding protein of the co-formulation of the invention contains a VL domain and a VH domain that have up to one, two, three, four or five or more amino acid substitutions and exhibits specific binding to PD-1.

[0232] In any of the above embodiments, the PD-1 API can be a full-length anti-PD-1 antibody or antigen-binding fragment thereof that specifically binds human PD-1. In certain embodiments, the PD-1 API is a full-length anti-PD-1 antibody selected from any class of immunoglobulin, including IgM, IgG, IgD, IgA, and IgE. Preferably, the antibody is an IgG antibody. Any isotype of IgG can be used, including IgG1, IgG2, IgG3, and IgG4. Different constant domains can be added to the VL and VH regions provided herein. For example, heavy chain constant domains other than IgG1 can be used if a particular intended use of the antibody (or fragment) of the invention requires altered effector function. IgG1 antibodies offer long half-life and effector functions (e.g., complement activation and antibody-dependent cellular cytotoxicity), but such activity may not be desirable for all applications of the antibody. In such cases, for example, an IgG4 constant domain can be used.

[0233] In an embodiment of the invention, the PD-1 API is an anti-PD-1 antibody comprising a light chain that comprises or consists of the sequence of amino acid residues set forth in SEQ ID NO:99, and a heavy chain that comprises or consists of the sequence of amino acid residues set forth in SEQ ID NO:104. In an alternative embodiment, the PD-1 API is an anti-PD-1 antibody comprising a light chain that comprises or consists of the sequence of amino acid residues set forth in SEQ ID NO:109, and a heavy chain that comprises or consists of the sequence of amino acid residues set forth in SEQ ID NO:114. In a further embodiment, the PD-1 API is an anti-PD-1 antibody comprising a light chain that comprises or consists of the sequence of amino acid residues set forth in SEQ ID NO:126, and a heavy chain that comprises or consists of the sequence of amino acid residues set forth in SEQ ID NO:31251. In a further embodiment, the PD-1 API is an anti-PD-1 antibody comprising a light chain that comprises or consists of the sequence of amino acid residues set forth in SEQ ID NO:127, and a heavy chain that comprises or consists of the sequence of amino acid residues set forth in SEQ ID NO:125. In yet additional embodiments, the PD-1 API is an anti-PD-1 antibody comprising a light chain that comprises, or consists of, the sequence of amino acid residues set forth in SEQ ID NO: 128 and a heavy chain that comprises, or consists of the sequence of amino acid residues set forth in SEQ ID NO: 125. In some co-formulations of the invention, the PD-1 API is pembrolizumab or a pembrolizumab biosimilar. In some co-formulations of the invention, the PD-1 API is nivolumab or a nivolumab biosimilar.

[0234] Typically, amino acid sequence variants of the anti-PD-1 antibodies and antigen-binding fragments, and anti-ILT4 antibodies and antigen-binding fragments of the invention will have an amino acid sequence that has at least 75% amino acid sequence identity, more preferably at least 80%, more preferably at least 85%, more preferably at least 90%, and most preferably at least 95%, 98% or 99% amino acid sequence identity to the amino acid sequence of a reference antibody or antigen-binding fragment (e.g., heavy chain, light chain, VH, VL, or humanized sequence). Identity or homology with respect to a sequence is defined herein as the percentage of amino acid residues in a candidate sequence that are identical to the anti-PD-1 residues after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, without considering conservative substitutions as part of the sequence identity. Any N-terminal, C-terminal or internal extensions, deletions, or insertions into the antibody sequence shall not be construed as affecting sequence identity or homology.

[0235] Sequence identity refers to the degree to which the amino acids of two polypeptides are identical at equivalent positions when the two sequences are optimally aligned.Sequence identity can be determined using the BLAST algorithm, where the parameters of the algorithm are selected to maximize the match between the respective sequences over the entire length of each reference sequence. The following references relate to the BLAST algorithm, which is often used for sequence analysis: BLAST ALGORITHMS: Altschul, SF, et al., (1990) J. Mol. Biol. 215:403-410; Gish, W., et al., (1993) Nature Genet. 3:266-272; Madden, TL, et al., (1996) Meth. Enzymol. 266:131-141; Altschul, SF, et al., (1997) Nucleic Acids Res. 25:3389-3402; Zhang, J., et al., (1997) Genome Res. 7:649-656; Wootton, JC, et al., (1993) Comput. Chem. 17:149-163; Hancock, JM et al. al., (1994) Comput. Appl. Biosci. 10:67-70; ALIGNMENT SCORING SYSTEMS: Dayhoff, MO, et al., ``A model of evolutionary change in proteins.'' in Atlas of Protein Sequence and Structure, (1978) vol. 5, suppl. 3. MO Dayhoff (ed.), pp. 345-352, Natl. Biomed. Res. Found., Washington, DC; Schwartz, RM, et al., ``Matrices for detecting distant relationships.'' in Atlas of Protein Sequence and Structure, (1978) vol. 5, suppl. 3.” MO Dayhoff (ed.), pp. 353-358, Natl. Biomed. Res. Found., Washington, DC; Altschul, SF, (1991) J. Mol. Biol. 219:555-565; States, DJ, et al., (1991) Methods 3:66-70; Henikoff, S., et al., (1992) Proc. Natl. Acad. Sci. USA 89:10915-10919; Altschul, SF, et al., (1993) J. Mol. Evol. 36:290-300; ALIGNMENT STATISTICS: Karlin, S., et al., (1990) Proc. Natl. Acad. Sci. USA 87:2264-2268; Karlin, S., et al., (1993) Proc. Natl. Acad. Sci. USA 90:5873-5877; Dembo, A., et al., (1994) Ann. Prob. 22:2022-2039; and Altschul, SF ``Evaluating the statistical significance of multiple distinct local alignments.'' in Theoretical and Computational Methods in Genome Research (S. Suhai, ed.), (1997) pp. 1-14, Plenum, New York. .

[0236] Similarly, any class of light chain can be used in the compositions and methods herein. In particular, kappa, lambda or variants thereof are useful in the compositions and methods of the invention.

[0237] The formulation further comprises an anti-PD-1 antibody or antigen-binding fragment, examples of which are provided below.

[0238] [Table 4] TIFF2025514174000006.tif103164

[0239] [Table 5]

[0240] As used herein, "pembrolizumab variant" refers to a monoclonal antibody comprising heavy and light chain sequences that are substantially identical to those in pembrolizumab, except that the heavy and light chain sequences have three, two or one conservative amino acid substitutions at positions outside the light chain CDRs and six, five, four, three, two or one conservative amino acid substitutions at positions outside the heavy chain CDRs, e.g., the variant positions are located in the FR regions or constant regions, and optionally the C-terminal lysine residue of the heavy chain is deleted. In other words, pembrolizumab and pembrolizumab variants contain identical CDR sequences but differ from each other by having conservative amino acid substitutions at only three or six other positions in their full-length light and heavy chain sequences, respectively. Pembrolizumab variants are substantially similar to pembrolizumab with respect to the following properties: binding affinity to PD-1 and ability to block the binding of each of PD-L1 and PD-L2 to PD-1.

[0241] In another embodiment of the invention, the formulation comprises an anti-ILT4 antibody or antigen-binding fragment comprising a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E or D and X2 is S or A) (SEQ ID NO: 20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21); and an anti-PD-1 antibody or antigen-binding fragment comprising a light chain variable region sequence of SEQ ID NO: 98 and a heavy chain variable region sequence of SEQ ID NO: 103. In another embodiment, the formulation comprises an anti-ILT4 antibody, wherein the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21); and an anti-PD-1 antibody comprising a light chain sequence of SEQ ID NO: 5 and a heavy chain sequence of SEQ ID NO: 104. The invention further provides formulations of an anti-ILT4 antibody or antigen-binding fragment thereof (a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:58); and an anti-PD-1 antibody comprising a light chain (CDRL1 sequence of SEQ ID NO:95, CDRL2 sequence of SEQ ID NO:96, and CDRL3 sequence of SEQ ID NO:97) and a heavy chain (CDRH1 sequence of SEQ ID NO:100, CDRH2 sequence of SEQ ID NO:101, and CDRH3 sequence of SEQ ID NO:102). In some formulations of the invention, the anti-ILT4 antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2; and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.In some embodiments, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:80; and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0242] In a further aspect of the invention, the formulation comprises an anti-ILT4 antibody, or antigen-binding fragment thereof, comprising a heavy chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:57 and a light chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:58, and an anti-PD-1 antibody, or antigen-binding fragment thereof, comprising a light chain variable region sequence of SEQ ID NO:108 and a heavy chain variable region sequence of SEQ ID NO:113. The invention further provides formulations of an anti-ILT4 antibody, or antigen-binding fragment thereof, comprising a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:80 and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7, and an anti-PD-1 antibody, or antigen-binding fragment thereof, comprising a light chain variable region (CDRL1 sequence of SEQ ID NO:105, CDRL2 sequence of SEQ ID NO:106, and CDRL3 sequence of SEQ ID NO:107) and a heavy chain variable region (CDRH1 sequence of SEQ ID NO:110, CDRH2 sequence of SEQ ID NO:111, and CDRH3 sequence of SEQ ID NO:112).

[0243] The antibody or antigen-binding fragment of the formulation can contain a light chain variable region and a heavy chain variable region. In some embodiments, the light chain variable region comprises SEQ ID NO:98 or a variant of SEQ ID NO:98, and the heavy chain variable region comprises SEQ ID NO:103 or a variant of SEQ ID NO:103. In further embodiments, the light chain variable region comprises SEQ ID NO:108 or a variant of SEQ ID NO:108, and the heavy chain variable region comprises SEQ ID NO:113 or a variant of SEQ ID NO:113. In further embodiments, the heavy chain variable region comprises SEQ ID NO:121 or a variant of SEQ ID NO:121, and the light chain variable region comprises SEQ ID NO:122 or a variant of SEQ ID NO:1228, SEQ ID NO:123 or a variant of SEQ ID NO:123, or a variant of SEQ ID NO:124 or a variant of SEQ ID NO:124. In such embodiments, the variant light or heavy chain variable region sequence is identical to the reference sequence except for having 1, 2, 3, 4 or 5 amino acid substitutions. In some embodiments, the substitutions are in the framework regions (i.e., outside the CDRs), hi some embodiments, 1, 2, 3, 4 or 5 of the amino acid substitutions are conservative substitutions.

[0244] In another embodiment, the formulation of the present invention comprises the V L Domain or V H V that shows at least 95%, 90%, 85%, 80%, 75% or 50% sequence identity to one of the domains L Domain and / or V H In another embodiment, the antibody or antigen-binding fragment of the formulation of the invention comprises an antibody or antigen-binding fragment having a V domain and exhibiting specific binding to PD-1 or ILT4. L Domain and V H domain and exhibits specific binding to PD-1 or ILT4.

[0245] In an embodiment of the invention, the antibody is an anti-PD-1 antibody comprising a light chain comprising or consisting of the sequence of amino acid residues set forth in SEQ ID NO:99 and a heavy chain comprising or consisting of the sequence of amino acid residues set forth in SEQ ID NO:104. In an alternative embodiment, the antibody is an anti-PD-1 antibody comprising a light chain comprising or consisting of the sequence of amino acid residues set forth in SEQ ID NO:109 and a heavy chain comprising or consisting of the sequence of amino acid residues set forth in SEQ ID NO:114. In a further embodiment, the antibody is an anti-PD-1 antibody comprising a light chain comprising or consisting of the sequence of amino acid residues set forth in SEQ ID NO:126 and a heavy chain comprising or consisting of the sequence of amino acid residues set forth in SEQ ID NO:125. In a further embodiment, the antibody is an anti-PD-1 antibody comprising a light chain comprising or consisting of the sequence of amino acid residues set forth in SEQ ID NO:127 and a heavy chain comprising or consisting of the sequence of amino acid residues set forth in SEQ ID NO:125. In a further embodiment, the antibody is an anti-PD-1 antibody comprising a light chain comprising, or consisting of, the sequence of amino acid residues set forth in SEQ ID NO: 128, and a heavy chain comprising, or consisting of, the sequence of amino acid residues set forth in SEQ ID NO: 125. In some formulations of the invention, the anti-PD-1 antibody is pembrolizumab or a pembrolizumab variant.

[0246] Typically, amino acid sequence variants of the anti-PD-1 or anti-ILT4 antibodies and antigen-binding fragments of the invention are similar to those of the reference antibody or antigen-binding fragment (e.g., heavy chain, light chain, V H , V LPreferably, the antibody will have an amino acid sequence that exhibits at least 75% amino acid sequence identity, more preferably at least 80%, more preferably at least 85%, even more preferably at least 90%, and most preferably at least 95%, 98% or 99% amino acid sequence identity with the amino acid sequence of the candidate sequence (anti-PD-1 or anti-ILT4 sequence, framework or humanized sequence). Identity or homology with respect to sequences is defined herein as the percentage of amino acid residues in the candidate sequence that are identical to the anti-PD-1 or anti-ILT4 residues after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percentage sequence identity, without considering conservative substitutions as part of the sequence identity. No N-terminal, C-terminal or internal extensions, deletions, or insertions into the antibody sequence shall be construed as affecting sequence identity or homology.

[0247] In one embodiment, the ratio of anti-ILT4 antibody to anti-PD-1 antibody in the formulation is 1:1, 1:2 or 1:3. In another embodiment, the molar ratio of anti-ILT4 antibody to anti-PD-1 antibody in the formulation is 1:1, 2:1, 3:1, 3.5:1, 4:1, 5:1 or 6:1. In one embodiment, the molar ratio of anti-ILT4 antibody to anti-PD-1 antibody in the formulation is 4:1. In another embodiment, the molar ratio of anti-ILT4 antibody to anti-PD-1 antibody in the formulation is 5:1.

[0248] formulation The co-formulations described herein can preserve the physical, chemical and / or biological stability of an anti-ILT4 antibody (e.g., 1E1, 2A6, 3G7 or 2C1) or an antigen-binding fragment thereof during storage (e.g., at about 5° C. for at least 6 months, 12 months, 24 months or 36 months) and / or under various stress conditions (e.g., agitation, freeze-thaw cycles or light exposure).

[0249] In one aspect, provided herein are various co-formulations of an anti-ILT4 antibody (e.g., 1E1, 2A6, 3G7, or 2C1) or an antigen-binding fragment thereof and an anti-PD-1 antibody or an antigen-binding fragment thereof, wherein the co-formulation comprises: (i) an anti-ILT4 antibody or an antigen-binding fragment thereof (e.g., 1E1, 2A6, 3G7, or 2C1); (ii) an anti-PD-1 antibody or an antigen-binding fragment thereof; (iii) a buffer (e.g., an L-histidine buffer or an acetate buffer); (iv) a non-reducing sugar (e.g., sucrose); (v) a non-ionic detergent (e.g., PS-80); and (vi) an antioxidant (e.g., L-methionine).

[0250] In some embodiments of the invention, the formulations of the invention minimize the formation of antibody aggregates (high molecular weight species) and particulates, improve colloidal stability, minimize fragmentation (low molecular weight species), or ensure that the antibody maintains its biological activity over time. In one embodiment, the formulation comprises: about 3-300 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof and about 3-300 mg / mL of an anti-PD-1 antibody or antigen-binding fragment thereof in a molar ratio (anti-ILT4 antibody to anti-PD-1 antibody or antigen-binding fragment thereof) of 4:1-5:1; histidine, aspartate, glutamine, glycine, proline, methionine, arginine or pharma- ceutical acceptable salts thereof, NaCl, KCl, LiCl, CaCl 2 , MgCl 2 , ZnCl 2 and FeCl 2wherein the total excipient concentration is about 10-1000 mM; and a buffer having a pH of about 5-8. In one embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof and the anti-PD-1 antibody or antigen-binding fragment thereof have a molar ratio of 4:1 (anti-ILT4 antibody to anti-PD-1 antibody or antigen-binding fragment thereof). In another embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof and the anti-PD-1 antibody or antigen-binding fragment thereof have a molar ratio of 5:1 (anti-ILT4 antibody to anti-PD-1 antibody or antigen-binding fragment thereof). In another aspect, the formulation comprises about 4-200 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof and about 4-200 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof. In one embodiment, histidine, aspartate, glutamine, glycine, proline, methionine, arginine or a pharma- ceutically acceptable salt thereof, NaCl, KCl, LiCl, CaCl 2 , MgCl 2 , ZnCl 2 and FeCl 2 In another embodiment, the one or more excipients selected from the group consisting of histidine, aspartate, glutamine, glycine, proline, methionine, arginine or pharma- ceutically acceptable salts thereof, NaCl, KCl, LiCl, CaCl, and the like, have a total excipient concentration of about 25 to 250 mM. 2 , MgCl 2 , ZnCl 2 and FeCl 2 The one or more excipients selected from the group consisting of have a total excipient concentration of about 40 to 250 mM.

[0251] In one embodiment, the excipient is arginine or a pharma- ceutically acceptable salt thereof at a concentration of about 15-250 mM. In one embodiment, the excipient is arginine or a pharma- ceutically acceptable salt thereof at a concentration of about 25-250 mM. In another embodiment, the excipient is arginine or a pharma- ceutically acceptable salt thereof at a concentration of about 40-150 mM. In another embodiment, the excipient is arginine or a pharma- ceutically acceptable salt thereof at a concentration of about 40-100 mM. In another embodiment, the excipient is L-arginine or a pharma- ceutically acceptable salt thereof at a concentration of about 70 mM. In another embodiment, the excipient is arginine or a pharma- ceutically acceptable salt thereof at a concentration of about 70-150 mM. Examples of pharma- ceutically acceptable salts of arginine (L- or D-form) include, but are not limited to, L-arginine hydrochloride and L-arginine succinate. In another aspect of the foregoing embodiment, the formulation further comprises a non-ionic surfactant, a sugar or a polyol, or glutamine, glycine, proline, or methionine.

[0252] In another aspect, the excipients are NaCl and arginine or a pharma- ceutically acceptable salt thereof, and the total excipient concentration is about 25-250 mM. In a further embodiment, the excipients are NaCl and arginine or a pharma- ceutically acceptable salt thereof, and the total excipient concentration is about 70-100 mM. In one embodiment, the concentration ratio of NaCl to arginine is 1:1. In another embodiment, the NaCl concentration is about 35 mM and the arginine concentration is about 35 mM. In another embodiment, the NaCl concentration is about 50 mM and the arginine concentration is about 50 mM.

[0253] In a further aspect, the excipient is about 40-150 mM NaCl, KCl or LiCl. In a further embodiment, the excipient is about 40-100 mM NaCl, KCl or LiCl. In a further embodiment, the excipient is about 70-130 mM NaCl, KCl or LiCl. In a further embodiment, the excipient is about 70-100 mM NaCl, KCl or LiCl. In a further embodiment, the excipient is about 70 mM NaCl. In another aspect of the foregoing embodiment, the formulation further comprises a non-ionic surfactant.

[0254] In a further aspect, the excipient is about 25-200 mM L-histidine. In a further embodiment, the L-histidine is about 50-200 mM. In a further embodiment, the L-histidine is about 40-100 mM.

[0255] In a further aspect, the excipient is about 25-200 mM L-glutamine, L-glycine, L-proline, or L-methionine, or a combination thereof. In a further embodiment, the excipient is about 50-200 mM. In a further embodiment, the excipient is about 40-100 mM. In a further embodiment, the excipient is about 70 mM.

[0256] In one embodiment, the excipient is about 25-200 mM L-glutamine, L-glycine, L-aspartate, or a combination thereof. In another embodiment, the excipient is about 20-50 mM. In a further embodiment, the excipient is about 20 mM. In a further embodiment, the excipient is about 40-100 mM. In a further embodiment, the excipient is about 70 mM. In another embodiment, the excipient is about 20 mM L-aspartate and about 50 mM L-glycine. In another embodiment, the excipient is about 20 mM L-glutamine and about 50 mM L-glycine.

[0257] In one embodiment, the co-formulated composition has a buffer having a neutral or slightly acidic pH (pH 4.5-8) and arginine or a pharma- ceutically acceptable salt thereof. In one embodiment, a buffer having a pH of about 5.5-6.5 is used in the composition. In one embodiment, a buffer having a pH of about 4.5-6.5 is used in the composition. In another embodiment, a buffer having a pH of about 5.5-6.0 is used in the composition. In a further embodiment, a buffer having a pH of about 5.0-6.0 is used in the composition. The buffer may have a concentration of about 5-1000 mM. In another embodiment, the buffer may have a concentration of about 5-150 mM. In a further embodiment, the buffer may have a concentration of about 5-300 mM. In a further embodiment, the buffer may have a concentration of about 1-300 mM. In another embodiment, the buffer may have a concentration of about 1-30 mM. In a further embodiment, the buffer may have a concentration of about 5-30 mM. In further embodiments, the buffering agent may have a concentration of about 5-20 mM. In further embodiments, the buffering agent may have a concentration of about 8-12 mM. In one embodiment, the buffering agent is histidine, acetate, or citrate. In some embodiments, the buffering agent comprises about 10 mM histidine, acetate, or citrate.

[0258] In one embodiment, the formulation comprises about 3-300 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof and about 3-300 mg / mL of an anti-PD-1 antibody or antigen-binding fragment thereof in a molar ratio of 4:1-5:1 (anti-ILT4 antibody to anti-PD-1 antibody, or antigen-binding fragment thereof), a sugar or polyol; a non-ionic surfactant, a histidine or acetate buffer of about pH 4.5-8, about 10-1000 mM methionine (L- or D-form) or a pharma- ceutically acceptable salt thereof, and optionally arginine, EDTA, DTPA, tryptophan (L- or D-form) or pyridoxine. In another embodiment, the formulation comprises about 4-250 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof and about 4-250 mg / mL of an anti-PD-1 antibody or antigen-binding fragment thereof in a molar ratio of 4:1-5:1 (anti-ILT4 antibody to anti-PD-1 antibody, or antigen-binding fragment thereof); a sugar or polyol; a non-ionic surfactant; about 50-500 mM histidine buffer at a pH of about 5-8; about 10-1000 mM of a salt of a monovalent cation selected from NaCl, KCl, and LiCl, or CaCl. 2 , MgCl 2 , ZnCl 2 , FeCl 2 and FeCl 3 and optionally about 10-1000 mM methionine (D or L) or a pharma- ceutically acceptable salt thereof, and optionally arginine, EDTA, DTPA, tryptophan, and pyridoxine. In another aspect, the formulation comprises about 4-200 mg / mL of an anti-ITL4 antibody or antigen-binding fragment thereof and about 4-200 mg / mL of an anti-PD-1 antibody or antigen-binding fragment thereof. In one embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof and the anti-PD-1 antibody or antigen-binding fragment thereof have a molar ratio of 4:1 (anti-ILT4 antibody to anti-PD-1 antibody or antigen-binding fragment thereof). In another embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof and the anti-PD-1 antibody or antigen-binding fragment thereof have a molar ratio of 5:1 (anti-ILT4 antibody to anti-PD-1 antibody or antigen-binding fragment thereof).

[0259] The formulation may contain about 1-100 uM, about 1-30 uM, about 1-20 uM, or about 10 uM-30 uM of DTPA or EDTA. The formulation may further contain about 1-30 mM L-methionine. In one embodiment, the formulation may further contain about 1-20 mM L-methionine. The formulation may further contain about 5-15 mM L-methionine. The formulation may further contain about 5-15 mM L-methionine. The formulation may further contain about 5-20 mM L-methionine. The formulation may further contain about 10 mM or at least about 10 mM L-methionine. In order to stabilize the antibody against oxidation, nitrogen overlay (blanketing, e.g., only 5% or 10% residual O during nitrogen overlay) may be used during the manufacturing process and / or prior to vial closure. 2 ) may also be used.

[0260] In another aspect of the invention, the formulation further comprises a sugar, a polyol or a non-ionic surfactant or a combination thereof. In one embodiment, the sugar is selected from the group consisting of glucose, sucrose, trehalose and lactose or a combination thereof. In one embodiment, the sugar is a disaccharide such as sucrose, trehalose and maltose. In one embodiment, the sugar is a non-reducing sugar. In another embodiment, the sugar is a non-reducing disaccharide such as sucrose, trehalose or a combination thereof. In one embodiment, the sugar is at a concentration of about 10-200 mg / mL. In another embodiment, the sugar is at a concentration of about 30-120 mg / mL. In another embodiment, the sugar is at a concentration of about 30-80 mg / mL. In a further embodiment, the sugar is at a concentration of about 50-90 mg / mL.

[0261] In one embodiment, the polyol is selected from the group consisting of mannitol, sorbitol, glycerol, and polyethylene glycol. In another embodiment, the polyol is a sugar alcohol. In one embodiment, the sugar and polyol are selected from the group consisting of sucrose, trehalose, sorbitol, glycerol, and polyethylene glycol. In a further embodiment, the polyol is a glycol. In one embodiment, the glycol is selected from the group consisting of ethylene glycol, propylene glycol, and polyethylene glycol. In one embodiment, the polyol is at a concentration of about 10-200 mg / mL. In another embodiment, the polyol is at a concentration of about 10-50 mg / mL. In a further embodiment, the polyol is at a concentration of about 5-30 mg / mL.

[0262] In one embodiment, the formulation comprises about 10-250 mg / mL sucrose or trehalose. In another embodiment, the formulation comprises about 20-200 mg / mL sucrose or trehalose. In a further embodiment, the formulation comprises about 50-80 mg / mL sucrose or trehalose. In a further embodiment, the formulation comprises about 30-80 mg / mL sucrose or trehalose. In another embodiment, the formulation comprises about 50-90 mg / mL sucrose or trehalose. In a further embodiment, the formulation comprises about 70-80 mg / mL sucrose or trehalose. In a further embodiment, the formulation comprises at least about 50 mg / mL sucrose or trehalose. In another embodiment, the formulation comprises about 20-200 mg / mL sorbitol, PEG400 or glycerol. In a further embodiment, the formulation comprises about 20-50 mg / mL of sorbitol, PEG400, or glycerol.

[0263] In one embodiment, the non-ionic surfactant is selected from the group consisting of polysorbates and poloxamers. In yet another embodiment, the surfactant is selected from the group consisting of Tween 80® (polysorbate 80), Tween 20® (polysorbate 20), Pluronic F88®, Pluoronic F-127®, Pluronic F68®, Triton X-100®. In one embodiment, the surfactant is polysorbate 20 or polysorbate 80, and the sugar is sucrose or trehalose. The polysorbate 80 or polysorbate 20 surfactant may be present in the formulation in an amount of about 0.005 to about 1 mg / mL. The polysorbate 80 or polysorbate 20 surfactant may be present in the formulation in an amount of about 0.02 to about 2 mg / mL. The polysorbate 80 or polysorbate 20 surfactant may be present in the formulation in an amount of about 0.05 to about 1 mg / mL. The polysorbate 80 or polysorbate 20 surfactant may be present in the formulation in an amount of about 0.1 to about 0.5 mg / mL. In another embodiment, the polysorbate 80 or polysorbate 20 surfactant may be present in the formulation in an amount of at least about 0.005 mg / mL. The polysorbate 80 or polysorbate 20 surfactant may further be present in the formulation in an amount of at least about 0.1 mg / mL. The polysorbate 80 surfactant may be present in the formulation in an amount of about 0.2 mg / mL.

[0264] In other embodiments of the above formulations, the % high molecular weight (HMW) in the co-formulated anti-ILT4 antibody and anti-PD-1 antibody formulation after 10 days at 50° C. is less than 5% as measured by size exclusion chromatography.

[0265] Liquid formulations Liquid antibody formulations can be produced by placing the drug substance, for example, in an aqueous pharmaceutical formulation and buffer exchanging it into a desired buffer as the final step in the purification process. The drug substance in the final buffer is concentrated to the desired concentration. Excipients such as stabilizers and surfactants are added to the drug substance and diluted with an appropriate buffer to the final protein concentration. The final formulated drug substance is filtered using a 0.22 μm filter and filled into the final container (e.g., glass vial). The formulation can be stored in the vial and delivered through an injection device or container.

[0266] In another aspect of the invention, the liquid co-formulation comprises an anti-PD-1 antibody and an anti-ILT4 antibody (or antigen-binding fragment thereof), wherein the anti-ILT4 antibody or antigen-binding fragment thereof has a concentration of about 10-300 mg / mL. In another embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof has a concentration of about 15-250 mg / mL. In another embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof has a concentration of about 20-100 mg / mL. In a further embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof has a concentration of about 30-80 mg / mL. In a further embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof has a concentration of about 35-75 mg / mL. In a further embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof has a concentration of about 40-70 mg / mL. In a further embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof has a concentration of about 45-65 mg / mL. In another embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof has a concentration of about 45-60 mg / mL. In yet another embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof has a concentration of about 45-65 or 55 mg / mL. In yet another embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof is at a concentration of about 40-60 mg / mL or 45-55 mg / mL. In a further embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof is at a concentration of about 46-52 mg / mL. In a further embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof is at a concentration of about 48-52 mg / mL. In a further embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof is at a concentration of about 49 mg / mL. In a further embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof is at a concentration of about 50 mg / mL. In a further embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof is at a concentration of about 51 mg / mL.

[0267] In another aspect of the invention, the anti-PD-1 antibody or antigen-binding fragment thereof in the liquid formulation has a concentration of about 3-300 mg / mL. In one embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof is at a concentration of about 4-250 mg / mL. In another embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof is at a concentration of about 40-100 mg / mL. In a further embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof is at a concentration of about 10-60 mg / mL. In a further embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof is at a concentration of about 20-30 mg / mL. In a further embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof is at a concentration of about 10-30 mg / mL. In a further embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof is at a concentration of about 15-50 mg / mL. In another embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof is at a concentration of about 10-100 mg / mL. In another embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof has a concentration of about 20-30 or 25 mg / mL. In one embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof has a concentration of about 21-29 mg / mL. In one embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof has a concentration of about 22-28 mg / mL. In one embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof has a concentration of about 24 mg / mL. In one embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof has a concentration of about 25 mg / mL. In one embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof has a concentration of about 26 mg / mL.

[0268] In one embodiment, the liquid formulation comprises a buffer having a pH of about 4.5-8, 5.0-6.5, 5.5-6.5, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, or 6.2, and arginine or a pharma- ceutically acceptable salt thereof. In one embodiment, the liquid formulation comprises a buffer having a pH of about 5-8. In one embodiment, the liquid formulation comprises a buffer having a pH of about 5.0-6.5. In one embodiment, the liquid formulation comprises a buffer having a pH of about 5.0-6.0. In another embodiment, the buffer is histidine. In another embodiment, the buffer is citrate or acetate. In further embodiments, the liquid formulation comprises an acetate buffer having a pH of about 5-8, 5.0-6.5, 5.5-6.5, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1 or 6.2, and arginine or a pharma- ceutically acceptable salt thereof.

[0269] The liquid antibody formulation of the present invention is suitable for parenteral administration, such as intravenous, intramuscular, intraperitoneal or subcutaneous injection, and is particularly suitable for subcutaneous injection.

[0270] Medication and Administration Toxicity is considered when selecting an appropriate dosage of a therapeutic agent such as a humanized anti-ILT4 antibody or anti-PD-1 antibody (or antigen-binding fragment thereof). The toxicity and therapeutic efficacy of the antibody composition administered alone or in combination with another immunosuppressant can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, to determine the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index, which can be expressed as the ratio of LD50 to ED50. Antibodies that exhibit high therapeutic indices are preferred. The data obtained from these cell culture assays and animal studies can be used to set a dosage range for use in humans. The dosage of such compounds is preferably within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage can vary within this range depending on the dosage form used and the route of administration utilized.

[0271] Suitable routes of administration include parenteral administration, such as intramuscular, intradermal, subcutaneous, intramedullary and intrathecal, direct intraventricular, intravenous, and intraperitoneal injections. Drugs can be administered in a variety of conventional ways, such as intraperitoneal, parenteral, intraarterial, or intravenous injections. Administration methods that require limited amounts of solution (e.g., subcutaneous administration) require lyophilized formulations that allow reconstitution at high concentrations.

[0272] Alternatively, the antibody can be administered locally rather than systemically, for example, by injecting the antibody directly into the pathogen-induced lesion characterized by immunopathology, often in a depot or sustained release formulation. Additionally, the antibody can be administered in a targeted drug delivery system, for example, in liposomes coated with tissue-specific antibody, which target the pathogen-induced lesion characterized by immunopathology, and the liposomes are targeted to and selectively taken up by the diseased tissue.

[0273] The choice of therapeutic dosing regimen depends on several factors, including the serum or tissue turnover rate of the entity, the level of symptoms, the immunogenicity of the entity, and the accessibility of the target cells in the biological matrix. Preferably, the dosing regimen maximizes the amount of therapeutic delivered to the patient consistent with an acceptable level of side effects. Thus, the amount of biologic delivered depends, in part, on the particular entity and the severity of the condition being treated. Guidance is available for selecting appropriate dosages of antibodies, cytokines, and small molecules.Citation: Wawrzynczak (1996) Antibody Therapy, Bios Scientific Pub. Ltd, Oxfordshire, UK; Kresina (ed.) (1991) Monoclonal Antibodies, Cytokines and Arthritis, Marcel Dekker, New York, NY; Bach (ed.) (1993) Monoclonal Antibodies and Peptide Therapy in Autoimmune Diseases, Marcel Dekker, New York, NY; Baert et al. (2003) New Engl. J. Med. 348:601-608; Milgrom et al. (1999) New Engl. J. Med. 341:1966-1973; Slamon et al. (2001) New Engl. J. Med. 344:783-792; Beniaminovitz et al. (2000) New Engl. J. Med. 342:613-619; Ghosh et al. (2003) New Engl. J. Med. 348:24-32; Lipsky et al. (2000) New Engl. J. Med. 343:1594-1602; Physicians' Desk Reference 2003 (Physicians' Desk Reference, 57th Ed); Medical Economics Company; ISBN: 1563634457; 57th edition (November 2002)。

[0274] The determination of the appropriate dosage is made by the clinician, for example, using parameters or factors known or suspected in the art that affect or are expected to affect treatment. The appropriate dosage of the protein ("therapeutically effective amount") depends, for example, on the condition being treated, the severity and course of the condition, whether the protein is administered for prophylactic or therapeutic purposes, previous treatments, the patient's clinical history and response to the protein, the type of protein used, and the discretion of the attending physician. Generally, the administration is started at an amount somewhat lower than the optimal dose, and then increased in small increments until a desired or optimal effect is achieved relative to negative side effects. Important diagnostic indicators include, for example, symptoms (e.g., symptoms of inflammation) or the level of inflammatory cytokines produced. The protein is suitably administered to the patient once or repeatedly. The protein can be administered alone or in combination with other drugs or therapies.

[0275] The antibody or antibody fragment can be provided by continuous infusion or by spaced doses, e.g., once daily, 1 to 7 times per week, once per week, once every two weeks, once every three weeks, once per month, every other month, etc. A preferred dosing protocol is one which involves the maximum dose or maximum dosing frequency that avoids significant undesirable side effects.

[0276] In certain embodiments, the pharmaceutical formulations of the present invention are administered by intravenous (IV) infusion or injection.

[0277] In another embodiment, the pharmaceutical formulation of the present invention is administered by subcutaneous administration. Subcutaneous administration can be performed by injection using a syringe or other injection device (e.g., Inject-ease® device); injection pen; or needle-free device (e.g., MediJector and BioJector®).

[0278] Subcutaneous administration can be performed by injection using a syringe, autoinjector, syringe pen or needleless injection device. Intravenous injection can be performed after dilution of the formulation with a suitable commercially available diluent such as saline or 5% dextrose in water.

[0279] Although the high concentration solution formulations of the present invention are particularly advantageous for applications requiring high concentrations of antibodies, there is no reason why the formulations cannot be used at lower concentrations in situations where high concentrations are not necessary or desirable. Low concentrations of antibodies may be useful for low dose subcutaneous administration or in other modes of administration where the deliverable volume is substantially greater than 1 mL (e.g., intravenous administration). Such low concentrations include about 15, 10, 5, 2, 1 mg / mL or less.

[0280] Purpose The present invention provides lyophilized or liquid formulations of anti-ILT4 antibodies, or antigen-binding fragments thereof, and anti-PD-1 antibodies, or antigen-binding fragments thereof, for use in the treatment of cancer and infectious diseases.

[0281] Those skilled in the art will understand that the term "cancer" is the name of a disease in which body cells become abnormal and divide uncontrollably.Cancers that may be treated by the compounds, compositions and methods of the present invention include, but are not limited to, the following: Heart: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma, and teratoma; Lung: bronchogenic carcinoma (squamous cell, small undifferentiated cell, large undifferentiated cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondroitin hamartoma, mesothelioma; Gastrointestinal tract: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (tubular adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, vipoma), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Karposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma), colorectum; genitourinary tract: kidney (adenocarcinoma, Wilms' tumor [nephroblastoma], lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma); Liver: hepatocellular carcinoma, cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Bone: osteogenic sarcoma, fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, osteochondroma (osteochondral exostosis), benign chondroma, chondroblastoma, chondromyxoid fibroma, osteoid osteoma, giant cell tumor; Nervous system: skull (osteoma, hemangioma, granuloma, xanthomas, osteitis deformans), meninges (meningioma, meningeal sarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma [pinealoma], glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal cord neurofibroma, meningioma, glioma, sarcoma);Gynecological: uterus (endometrial carcinoma), cervix (cervical carcinoma, preneoplastic cervical dysplasia), ovary (ovarian carcinoma [serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma], granulosa theca cell tumor, Sertoli-Leydig cell tumor, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, carcinoma in situ, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma), fallopian tube (carcinoma), breast; Hematological: blood (myeloid leukemia [acute and chronic], acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative disorders, multiple myeloma, myelodysplastic syndromes), Hodgkin's disease, non-Hodgkin's lymphoma [malignant lymphoma]; Skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Karposi's sarcoma, lenticular dysplasia, lipoma, hemangioma, dermatofibroma, keloid, psoriasis; and Adrenal: neuroblastoma. In one embodiment, the cancer is selected from colorectal cancer, gastric cancer, and head and neck cancer.

[0282] Buffering agents that can be used in the pharmaceutical formulations disclosed herein include, but are not limited to, succinate (sodium or potassium), L-histidine, phosphate (sodium or potassium), Tris (tris(hydroxymethyl)aminomethane), diethanolamine, citrate (sodium), acetate (sodium), etc. In some embodiments of the formulations, the buffering agent is present in the formulation at a concentration of about 1-20 mM (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mM).

[0283] The buffers of the various formulations described herein have a pH range of about 4.5 to about 7.0, about 4.5 to about 6.8, about 5.0 to about 6.8, about 5.0 to about 6.5, about 5.0 to about 6.0, and about 5.5 to about 6.0. In arriving at a representative formulation, L-histidine and acetate buffers in the pH range of 5.0 to 6.8 were examined for suitability. When a range of pH values ​​is described, such as "a pH of about pH 5.5 to about 6.0", the range is intended to include the described value. For example, the range of about 5.0 to about 6.0 includes 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, and 6.0. When the anti-ILT4 formulation described herein is a lyophilized formulation, the pH refers to the pH after reconstitution of the lyophilized formulation. pH is typically measured with a standard glass bulb pH meter at 25° C. As used herein, a solution containing "a histidine buffer of pH X" refers to a solution containing a histidine buffer at pH X, i.e., the pH is intended to refer to the pH of the solution.

[0284] In some embodiments, the anti-ILT4 formulation comprises a non-reducing sugar. As used herein, a "non-reducing sugar" is a sugar that does not contain or cannot be converted to contain a free aldehyde or ketone group and therefore cannot act as a reducing agent. Examples of non-reducing sugars include, but are not limited to, disaccharides such as sucrose and trehalose. In an embodiment of the invention, the non-reducing sugar is present in an amount of about 1% (w / v) to about 10% (w / v) (about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10% (w / v)). In another embodiment, the non-reducing sugar is present in an amount of about 6% (w / v) to about 8% (w / v) (about 6, 7, or 8% (w / v)). In a further embodiment, the non-reducing sugar is present in an amount of about 6% (w / v). In a further embodiment, the non-reducing sugar is present in an amount of about 7% (w / v). In a further embodiment, the non-reducing sugar is present in an amount of about 8% (w / v). In one embodiment, the non-reducing sugar is sucrose, trehalose or raffinose. In a further embodiment, sucrose is present in an amount of about 6% (w / v) to about 8% (w / v). In one embodiment, sucrose is present at about 6% (w / v). In one embodiment, sucrose is present at about 7% (w / v). In one embodiment, sucrose is present at about 8% (w / v).

[0285] In certain embodiments, the anti-ILT4 formulation further comprises a surfactant. As used herein, a "surfactant" is a surface active agent that is amphiphilic in nature. Surfactants can be added to the formulations herein to provide stability, reduce and / or prevent aggregation, or prevent and / or inhibit protein damage under process conditions such as purification, filtration, lyophilization, transportation, storage and delivery. In some embodiments, surfactants can be useful to provide additional stability to the active ingredient(s).

[0286] Nonionic surfactants that can be used in the formulations of the invention include, but are not limited to, polyoxyethylene sorbitan fatty acid esters (polysorbates, sold under the trade name Tween® (Uniquema Americas LLC, Wilmington, DE)), such as polysorbate-20 (PS-20, polyoxyethylene sorbitan monolaurate), polysorbate-40 (PS-40, polyoxyethylene sorbitan monopalmitate), polysorbate-60 (PS-60, polyoxyethylene sorbitan monostearate), and polysorbate-80 (PS-80, polyoxyethylene sorbitan monooleate); polyoxyethylene alkyl ethers, such as Brij® 58 (Uniquema Americas LLC, Wilmington, DE) and Brij® 35; poloxamers (e.g., poloxamer 188); Triton® X-100 (Union Carbide Corp., Houston, TX) and Triton® X-114; NP40; Span 20, Span 40, Span 60, Span 65, Span 80 and Span 85; copolymers of ethylene and propylene glycol (e.g., the pluronic® series of nonionic surfactants, such as pluronic® F68, pluronic® 10R5, pluronic® F108, pluronic® F127, pluronic® F38, pluronic® L44, pluronic® L62 (BASF Corp., Ludwigshafen, Germany); and sodium dodecyl sulfate (SDS). In one embodiment, the nonionic surfactant is PS-80 or PS-20. In one embodiment, the nonionic surfactant is PS-20. In another embodiment, the nonionic surfactant is PS-80.

[0287] The amount of non-ionic surfactant included in the formulation is sufficient to perform the desired function, i.e., the minimum amount required to stabilize the active pharmaceutical ingredient (i.e., anti-ILT4 antibody or antigen-binding fragment thereof (e.g., 1E1, 2A6, 3G7, or 2C1)) in the formulation. All percentages of the non-ionic surfactant are listed in % (w / v). Typically, the surfactant is present at a concentration of about 0.008% to about 0.1% (w / v). In some embodiments of this aspect of the invention, the surfactant is present in the formulation in an amount of about 0.01% to about 0.1%, about 0.01% to about 0.09%, about 0.01% to about 0.08%, about 0.01% to about 0.07%, about 0.01% to about 0.06%, about 0.01% to about 0.05%, about 0.01% to about 0.04%, about 0.01% to about 0.03%, about 0.01% to about 0.02%, about 0.015% to about 0.04%, about 0.015% to about 0.03%, about 0.015% to about 0.02%, about 0.02% to about 0.04%, about 0.02% to about 0.035%, or about 0.02% to about 0.03%. In alternative embodiments, the surfactant is present in an amount of about 0.01%, about 0.015%, about 0.02%, about 0.025%, about 0.03%, about 0.035% or about 0.04%.

[0288] In certain embodiments, the formulations described herein comprise about 0.01% to about 0.04% (w / v) PS-80. In one embodiment, the formulations described herein comprise PS-80 in an amount of about 0.01%. In one embodiment, the amount of PS-80 is about 0.015%. In another embodiment, the amount of PS-80 is about 0.02%. In a further embodiment, the amount of PS-80 is about 0.025%. In another embodiment, the amount of PS-80 is about 0.03%. In a further embodiment, the amount of PS-80 is about 0.035%. In another embodiment, the amount of PS-80 is about 0.04%. In a further embodiment, the amount of PS-80 is about 0.045%.

[0289] The formulations described herein further comprise methionine or a pharma- ceutically acceptable salt thereof as an antioxidant. In one embodiment, the methionine is L-methionine. In another embodiment, the methionine is a pharma- ceutically acceptable salt of L-methionine, e.g., methionine HCl. In one embodiment of the invention, methionine is present in the formulation at a concentration of about 1 to 20 mM (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 mM). In another embodiment, methionine is present at about 5 mM to about 10 mM (5, 6, 7, 8, 9, and 10 mM). In another embodiment, methionine is present at about 10 mM.

[0290] Accordingly, in one aspect, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 10 mg / mL to about 200 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 5 mM to about 20 mM of a buffer; (iii) about 6% to about 8% weight / volume (w / v) of a non-reducing sugar; (iv) about 0.01% to about 0.10% (w / v) of a non-ionic surfactant; and (v) about 1 mM to about 20 mM of an antioxidant.

[0291] In certain embodiments, the buffer is selected from the group consisting of an L-histidine buffer, an acetate buffer, and a citrate buffer. In one embodiment, the buffer is an L-histidine buffer. In another embodiment, the buffer is an acetate buffer. In yet another embodiment, the buffer is a citrate buffer.

[0292] In some embodiments, the non-reducing sugar is sucrose.

[0293] In certain embodiments, the non-ionic surfactant is PS-80 or PS-20. In one embodiment, the non-ionic surfactant is PS-80. In another embodiment, the non-ionic surfactant is PS-20.

[0294] In some embodiments, the antioxidant is L-methionine.

[0295] Accordingly, in another aspect, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 10 mg / mL to about 200 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 5 mM to about 20 mM L-histidine buffer; (iii) about 6% to about 8% (w / v) sucrose; (iv) about 0.01% to about 0.10% (w / v) PS-80; and (v) about 1 mM to about 20 mM L-methionine.

[0296] In some embodiments, the formulation comprises about 8 mM to about 12 mM L-histidine buffer.

[0297] In certain embodiments, the formulation comprises about 5 mM to about 10 mM L-methionine.

[0298] In another embodiment, the formulation comprises about 0.01% to about 0.05% (w / v) PS-80.

[0299] In yet another embodiment, the formulation comprises about 10 mg / mL to about 150 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 10 mg / mL, about 12.5 mg / mL, about 15 mg / mL, about 25 mg / mL, about 50 mg / mL, about 75 mg / mL, about 100 mg / mL, about 125 mg / mL or about 150 mg / mL. In one embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 10 mg / mL. In another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 12.5 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 15 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 25 mg / mL. In one embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 50 mg / mL. In another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 75 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 100 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 125 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 150 mg / mL.

[0300] Thus, in one particular embodiment, the formulation comprises about 25 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0301] In another specific embodiment, the formulation comprises about 50 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0302] In yet another specific embodiment, the formulation comprises about 75 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0303] In yet another specific embodiment, the formulation comprises about 100 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0304] In yet another specific embodiment, the formulation comprises about 125 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0305] In certain embodiments of the various formulations provided herein, the formulation has a pH of about 5.0 to about 6.8. In some embodiments, the formulation has a pH of about 5.5 to about 6.0. In other embodiments, the formulation has a pH of about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, or about 6.0. In one embodiment, the formulation has a pH of about 5.5. In another embodiment, the formulation has a pH of about 5.6. In yet another embodiment, the formulation has a pH of about 5.7. In yet another embodiment, the formulation has a pH of about 5.8. In another embodiment, the formulation has a pH of about 5.9. In yet another embodiment, the formulation has a pH of about 6.0.

[0306] Thus, in one particular embodiment, the pharmaceutical formulation of an anti-ILT4 antibody or antigen-binding fragment thereof comprises: (i) about 50 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer, pH about 5.5; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) polysorbate 80; and (v) about 10 mM L-methionine.

[0307] In certain embodiments of the various formulations disclosed herein, the anti-ILT4 antibody or antigen-binding fragment thereof is an anti-ILT4 antibody or antigen-binding fragment thereof described herein (e.g., 1E1, 2A6, 3G7, or 2C1). In some embodiments of the formulation, the anti-ILT4 antibody or antigen-binding fragment thereof is 1E1 or a variant thereof. In some embodiments of the formulation, the anti-ILT4 antibody or antigen-binding fragment thereof is 2A6 or a variant thereof. In some embodiments of the formulation, the anti-ILT4 antibody or antigen-binding fragment thereof is 3G7 or a variant thereof. In some embodiments of the formulation, the anti-ILT4 antibody or antigen-binding fragment thereof is 2C1 or a variant thereof.

[0308] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX 1 X 2 NRPS (where X 1 is N, Q, E or D, and X 2 a light chain variable domain comprising CDR-L2: QSFDNSLSAYV (SEQ ID NO: 21); and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 22).

[0309] In some embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0310] In another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57, and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:58.

[0311] In yet another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2, and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0312] In yet another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:80, and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0313] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:57, and a light chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:58.

[0314] In some embodiments of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:2, and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7.

[0315] In another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:80, and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7.

[0316] Accordingly, in some embodiments, provided herein is a pharmaceutical composition for an anti-ILT4 antibody or antigen-binding fragment thereof, comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine. and a pharmaceutical formulation, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a heavy chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX 1 X 2 NRPS (where X 1 is N, Q, E or D, and X 2 a light chain variable domain comprising CDR-L2: QSFDNSLSAYV (SEQ ID NO: 21); and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 22).

[0317] In certain embodiments, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof is The original binding fragment comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52) and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0318] In another embodiment, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57 and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:58.

[0319] In yet another embodiment, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2 and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0320] In yet another embodiment, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:80 and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0321] In some embodiments, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:57 and a light chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:58.

[0322] In some embodiments, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:2 and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7.

[0323] In some embodiments, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:80 and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7.

[0324] In one embodiment, the formulation is a liquid formulation. In another embodiment, the liquid formulation is stored at about 3° C. to about 5° C. In yet another embodiment, the liquid formulation is frozen at at least −70° C. or below. In yet another embodiment, the liquid formulation is a reconstituted solution from a lyophilized formulation.

[0325] In certain embodiments, after storage of the formulation at about 3° C. to about 5° C. for 6 months, (i) the % monomer of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 99% as measured by ultra-performance size exclusion chromatography; (ii) the OD 0.350-500(iii) the % main peak of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 63%, the % acidic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 23%, and / or the % basic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 14% as measured by high performance ion exchange chromatography; (iv) the number of subvisible particles of particles at least 2 μm in size is at most about 3500 as measured by microflow imaging; and / or (v) the % oxidation of one or more amino acid residues selected from the group consisting of W7, W102, M253, M359 and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% as measured by reduced peptide mapping.

[0326] In some embodiments, the % monomer of the anti-ILT4 antibody, or antigen-binding fragment thereof, is at least about 99% when measured by ultra performance size exclusion chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months.

[0327] In some embodiments, the formulation is stored at about 3° C. to about 5° C. for 6 months and then the OD 350-500 When measured at 100° C., the turbidity of the formulation is at most about 0.135.

[0328] In some embodiments, the % major peak of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 63% when measured by high performance ion exchange chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months. In some embodiments, the % acidic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 23% when measured by high performance ion exchange chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months. In some embodiments, the % basic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 14% when measured by high performance ion exchange chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months. In some embodiments, after storage of the formulation at about 3° C. to about 5° C. for 6 months, the % major peak of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 63%, the % acidic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 23%, and the % basic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 14% when measured by high performance ion exchange chromatography.

[0329] In some embodiments, after storage of the formulation at about 3° C. to about 5° C. for 6 months, the number of sub-visible particles having a size of at least 2 μm is up to about 3500 as measured by microflow imaging.

[0330] In some embodiments, the % oxidation of one or more amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% when measured by reduced peptide mapping after storage of the formulation at about 3° C. to about 5° C. for 6 months. In one embodiment, the % oxidation of one amino acid residue selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% when measured by reduced peptide mapping after storage of the formulation at about 3° C. to about 5° C. for 6 months. In one embodiment, when the formulation is stored at about 3° C. to about 5° C. for 6 months and then measured by reduced peptide mapping, the % oxidation rate of two amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4%. In one embodiment, when the formulation is stored at about 3° C. to about 5° C. for 6 months and then measured by reduced peptide mapping, the % oxidation rate of three amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4%. In one embodiment, when the formulation is stored at about 3° C. to about 5° C. for 6 months and then measured by reduced peptide mapping, the % oxidation rate of four amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4%. In one embodiment, when the formulation is stored at about 3° C. to about 5° C. for 6 months and then measured by reduced peptide mapping, the % oxidation rate of all five amino acid residues in the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4%.

[0331] The formulations described herein can be produced as liquid formulations. Liquid antibody formulations can be produced by buffer exchange of the drug substance (e.g., anti-ILT4 monoclonal antibody) in liquid form (e.g., anti-ILT4 monoclonal antibody in an aqueous pharmaceutical formulation) into a desired buffer as the final step in the purification process. In this embodiment, there is no lyophilization step. The drug substance in the final buffer is concentrated to the desired concentration. Excipients such as sucrose and PS80 are added to the drug substance and diluted with an appropriate buffer to the final protein concentration. The final formulated drug substance is filtered using a 0.22 μm filter and filled into the final container (e.g., glass vial).

[0332] The formulations described herein can also be reconstituted from lyophilized formulations. Lyophilized formulations of therapeutic proteins offer several advantages. Lyophilized formulations generally offer better chemical stability than solution formulations, and therefore have increased half-life. Lyophilized formulations can also be reconstituted at different concentrations depending on clinical factors such as the route of administration or dosing. For example, lyophilized formulations are reconstituted at higher concentrations (i.e., smaller volumes) if subcutaneous administration is required, or at lower concentrations if administered intravenously. Higher concentrations may be necessary if a higher dose is required for a particular subject, particularly for subcutaneous administration where the injection volume must be minimized. One such lyophilized antibody formulation is disclosed in U.S. Pat. No. 6,267,958, which is incorporated herein by reference in its entirety. Another lyophilized formulation of a therapeutic protein is disclosed in U.S. Pat. No. 7,247,707, which is incorporated herein by reference in its entirety.

[0333] Typically, lyophilized formulations are prepared with the intention of reconstituting a high concentration of drug product (DP, in an exemplary embodiment, an anti-ILT4 antibody or antigen-binding fragment thereof), i.e., with a low volume of water. Subsequent dilution with water or an isotonic buffer can then be easily used to reduce the DP to a low concentration. Typically, excipients are included in the lyophilized formulations of the invention at levels that result in a near-isotonic formulation when reconstituted at a high DP concentration, e.g., for subcutaneous administration. Reconstitution with a large amount of water to reduce the DP concentration necessarily reduces the tonicity of the reconstituted solution, but such reduction may be of little importance for non-subcutaneous administration (e.g., intravenous administration). If isotonicity is desired at a lower DP concentration, the lyophilized powder can be reconstituted with a standard low volume of water and then diluted with an isotonic diluent, such as 0.9% sodium chloride.

[0334] The lyophilized formulations of the present invention are formed by lyophilizing the pre-lyophilization solution. Lyophilization is accomplished by freezing the formulation followed by sublimation of water at a temperature suitable for primary drying. Under these conditions, the product temperature is below the eutectic or collapse temperature of the formulation. Typically, the shelf temperature for primary drying ranges from about -30 to 25°C at an appropriate pressure, typically in the range of about 50 to 250 mTorr, provided that the product remains frozen during primary drying. The formulation, the size and type of container (e.g., glass vial) in which the sample is placed, and the amount of liquid determine the time required for drying, which can range from a few hours to several days (e.g., 40 to 60 hours). The secondary drying step can be carried out at about 0 to 40°C, depending primarily on the type and size of the container and the type of protein used. The secondary drying time is determined by the desired residual moisture level in the product and typically takes at least about 5 hours. Typically, the moisture content of the lyophilized formulation is less than about 5%, preferably less than about 3%. The pressure can be the same as that employed in the primary drying step. The lyophilization conditions can vary depending on the formulation and vial size.

[0335] In some cases, to avoid transfer steps, it may be desirable to lyophilize the protein formulation in the container in which the protein reconstitution will be performed, e.g., a 3, 5, 10, 20, 50, or 100 cc vial.

[0336] The lyophilized formulation is reconstituted prior to administration. The protein may be reconstituted at a concentration of about 10, 15, 20, 25, 30, 40, 50, 60, 75, 80, 90, or 100 mg / mL, or at a higher concentration up to about 500 mg / mL, for example, 150 mg / mL, 200 mg / mL, 250 mg / mL, or 300 mg / mL. In one embodiment, the protein concentration after reconstitution is about 10-300 mg / mL. In one embodiment, the protein concentration after reconstitution is about 20-250 mg / mL. In one embodiment, the protein concentration after reconstitution is about 150-250 mg / mL. In one embodiment, the protein concentration after reconstitution is about 180-220 mg / mL. In one embodiment, the protein concentration after reconstitution is about 50-150 mg / mL. In one embodiment, the protein concentration after reconstitution is about 150 mg / mL. In one embodiment, the protein concentration after reconstitution is about 125 mg / mL. In one embodiment, the protein concentration after reconstitution is about 100 mg / mL. In one embodiment, the protein concentration after reconstitution is about 75 mg / mL. In one embodiment, the protein concentration after reconstitution is about 50 mg / mL. In one embodiment, the protein concentration after reconstitution is about 25 mg / mL. High protein concentrations are particularly useful when subcutaneous delivery of the reconstituted formulation is intended. However, for other routes of administration, such as intravenous administration, lower concentrations of protein may be desired (e.g., about 5-50 mg / mL).

[0337] Reconstitution is usually carried out at a temperature of about 25° C. to ensure complete hydration, although other temperatures may be used if desired. The time required for reconstitution depends, for example, on the type of diluent, the amount of excipients, and the protein. Exemplary diluents include sterile water, bacteriostatic water for injection (BWFI), a pH buffered solution (e.g., phosphate buffered saline), sterile saline, Ringer's solution, or dextrose solution.

[0338] How to use In another aspect, provided herein is a method for treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of any formulation described herein. In some specific embodiments of this method, the formulation is administered to the subject via intravenous administration. In another embodiment, the formulation is administered to the subject by subcutaneous administration. In one embodiment, the invention encompasses a method for treating cancer in a human patient, comprising administering to the patient any formulation described herein.

[0339] In any of the methods of the present invention, the cancer may be selected from the group consisting of melanoma, lung cancer, head and neck cancer, bladder cancer, breast cancer, gastrointestinal cancer, multiple myeloma, hepatocellular carcinoma, lymphoma, renal cancer, mesothelioma, ovarian cancer, esophageal cancer, anal cancer, biliary tract cancer, colorectal cancer, cervical cancer, thyroid cancer, salivary gland cancer, prostate cancer (e.g., hormone refractory prostate adenocarcinoma), pancreatic cancer, colon cancer, esophageal cancer, liver cancer, thyroid cancer, glioblastoma, glioma, and other neoplastic malignancies.

[0340] In some embodiments, the lung cancer is non-small cell lung cancer.

[0341] In an alternative embodiment, the lung cancer is small cell lung cancer.

[0342] In some embodiments, the lymphoma is Hodgkin's lymphoma.

[0343] In another embodiment, the lymphoma is non-Hodgkin's lymphoma. In a particular embodiment, the lymphoma is mediastinal large B-cell lymphoma.

[0344] In some embodiments, the breast cancer is triple-negative breast cancer.

[0345] In a further embodiment, the breast cancer is ER+ / HER2- breast cancer.

[0346] In some embodiments, the bladder cancer is urothelial carcinoma.

[0347] In some embodiments, the head and neck cancer is nasopharyngeal carcinoma. In some embodiments, the cancer is thyroid cancer. In other embodiments, the cancer is salivary gland cancer. In other embodiments, the cancer is squamous cell carcinoma of the head and neck.

[0348] In some embodiments, the cancer is a solid tumor with high levels of microsatellite instability (MSI-H).

[0349] In some embodiments, the cancer is a solid tumor with a high mutational burden.

[0350] In some embodiments, the cancer is metastatic colorectal cancer with high levels of microsatellite instability (MSI-H).

[0351] In some embodiments, the cancer is selected from the group consisting of colorectal cancer, esophageal cancer, melanoma, non-small cell lung cancer, ovarian cancer, renal cell carcinoma, and small cell lung cancer.

[0352] In another embodiment of the above method of treatment, the cancer is a hematological malignancy. In a particular embodiment, the hematological malignancy is acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), diffuse large B-cell lymphoma (DLBCL), EBV-positive DLBCL, primary mediastinal large B-cell lymphoma, T-cell / histiocyte-rich large B-cell lymphoma, follicular lymphoma, Hodgkin's lymphoma (HL), mantle cell lymphoma (MCL), multiple myeloma (MM), myeloid cell leukemia 1 protein (Mcl-1), myelodysplastic syndrome (MDS), non-Hodgkin's lymphoma (NHL) or small lymphocytic lymphoma (SLL).

[0353] Malignant tumors that show improved disease-free and overall survival rates associated with the presence of tumor-infiltrating lymphocytes in biopsy or surgical specimens, such as melanoma, colorectal cancer, liver cancer, renal cancer, gastric / esophageal cancer, breast cancer, pancreatic cancer and ovarian cancer, are included in the methods and treatments described herein. Subtypes of such cancers are known to be susceptible to immune control by T lymphocytes. In addition, refractory or recurrent malignant tumors whose growth can be inhibited using the antibodies described herein are also included.

[0354] Additional cancers that may benefit from treatment with the formulations described herein include cancers associated with persistent infection with viruses such as human immunodeficiency virus, hepatitis A, B and C viruses, Epstein-Barr virus, human papilloma virus, etc., known to be causally associated with Kaposi's sarcoma, liver cancer, nasopharyngeal carcinoma, lymphoma, cervical cancer, vulvar cancer, anal cancer, penile cancer and oral cancer.

[0355] The formulations can also be used to prevent or treat infectious diseases and infectious disorders. Thus, the present invention provides a method of treating a chronic infectious disease in a mammalian subject, comprising administering to the subject an effective amount of a formulation of the present invention. In certain embodiments of the method, the formulation is administered to the subject via intravenous administration. In another embodiment, the formulation is administered to the subject by subcutaneous administration.

[0356] These agents can be used alone or in combination with vaccines to stimulate immune responses against pathogens, toxins and self-antigens. Antibodies or antigen-binding fragments thereof can be used to stimulate immune responses against viruses that infect humans, including but not limited to human immunodeficiency virus, hepatitis A, B and C viruses, Epstein-Barr virus, human cytomegalovirus, human papilloma virus and herpes viruses. Antagonist anti-PD-1 antibodies or antibody fragments can be used to stimulate immune responses against infections by bacterial or fungal parasites and other pathogens. Viral infections by hepatitis B and C viruses and HIV are considered chronic viral infections.

[0357] The formulations of the invention can be administered to a patient in combination with one or more "additional therapeutic agents." The additional therapeutic agents can be biologic therapeutic agents (including, but not limited to, antibodies against VEGF, EGFR, Her2 / neu, VEGF receptors, other growth factor receptors, CD20, CD40, CD-40L, OX-40, 4-1BB, and ICOS), immunogenic agents (e.g., attenuated cancer cells, tumor antigens, antigen-presenting cells, e.g., dendritic cells pulsed with tumor-derived antigens or nucleic acids, immunostimulatory cytokines (e.g., IL-2, IFNα2, GM-CSF), and cells transfected with genes encoding immunostimulatory cytokines (e.g., but not limited to, GM-CSF).

[0358] As mentioned above, in some embodiments of the method of the present invention, the method further comprises administering an additional therapeutic agent. In certain embodiments, the additional therapeutic agent is an anti-PD-1 antibody or an antigen-binding fragment thereof, an anti-PD-L1 antibody or an antigen-binding fragment thereof, an anti-ILT4 antibody or an antigen-binding fragment thereof, an anti-ILT4 antibody or an antigen-binding fragment thereof, an anti-GITR antibody or an antigen-binding fragment thereof, an anti-CTL4 antibody or an antigen-binding fragment thereof, or an anti-CD27 antibody or an antigen-binding fragment thereof. In one embodiment, the additional therapeutic agent is a Newcastle Disease Virus vector expressing IL-12. In a further embodiment, the additional therapeutic agent is dinaciclib. In a further embodiment, the additional therapeutic agent is a STING agonist.

[0359] Suitable routes of administration can include, for example, parenteral delivery, e.g., intramuscular, subcutaneous and intrathecal, direct intracerebroventricular, intravenous, intraperitoneal, etc. Drugs can be administered in a variety of conventional ways, such as intraperitoneal, parenteral, intraarterial or intravenous injection. Administration methods that require limited solution volume (e.g., subcutaneous administration) require lyophilized formulations that allow reconstitution at high concentrations.

[0360] The choice of dosage of the additional therapeutic agent depends on several factors, including the serum or tissue turnover rate of the entity, the level of symptoms, the immunogenicity of the entity, and the accessibility of the target cells, tissues, or organs in the individual being treated. The dosage of the additional therapeutic agent should be an amount that produces an acceptable level of side effects. Thus, the dosage and frequency of administration of each additional therapeutic agent (e.g., biological or chemotherapeutic agent) will depend in part on the particular therapeutic agent, the severity of the cancer being treated, and the characteristics of the patient. Guidance is available for selecting appropriate dosages of antibodies, cytokines, and small molecules.See, for example: Wawrzynczak (1996) Antibody Therapy, Bios Scientific Pub. Ltd, Oxfordshire, UK; Kresina (ed.) (1991) Monoclonal Antibodies, Cytokines and Arthritis, Marcel Dekker, New York, NY; Bach (ed.) (1993) Monoclonal Antibodies and Peptide Therapy in Autoimmune Diseases, Marcel Dekker, New York, NY; Baert et al. (2003) New Engl. J. Med. 348:601-608; Milgrom et al. (1999) New Engl. J. Med. 341:1966-1973; Slamon et al. (2001) New Engl. J. Med. 344:783-792; Beniaminovitz et al. (2000) New Engl. J. Med. 342:613-619; Ghosh et al. (2003) New Engl. J. Med. 348:24-32; Lipsky et al. (2000) New Engl. J. Med. 343:1594-1602; Physicians' Desk Reference 2003 (Physicians' Desk Reference, 57th Ed); Medical Economics Company; ISBN: 1563634457; 57th edition (November 2002). The determination of an appropriate dosing regimen can be made by the clinician using, for example, parameters or factors known or suspected in the art to affect or be expected to affect treatment, and depends, for example, on the patient's clinical history (e.g., previous treatments), the type and stage of the cancer being treated, and biomarkers of response to one or more therapeutic agents in the combination therapy.

[0361] To facilitate the selection of pharma- ceutically acceptable carriers or excipients for additional therapeutic agents, various references are available. See, for example: Remington's Pharmaceutical Sciences and US Pharmacopeia: National Formulary, Mack Publishing Company, Easton, PA (1984); Hardman et al. (2001) Goodman and Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill, New York, NY; Gennaro (2000) Remington: The Science and Practice of Pharmacy, Lippincott, Williams, and Wilkins, New York, NY; Avis et al. (eds.) (1993) Pharmaceutical Dosage Forms: Parenteral Medications, Marcel Dekker, NY; Lieberman, et al. (eds.) (1990) Pharmaceutical Dosage Forms: Tablets, Marcel Dekker, NY; Lieberman et al. (eds.) (1990) Pharmaceutical Dosage Forms: Disperse Systems, Marcel Dekker, NY; Weiner and Kotkoskie (2000) Excipient Toxicity and Safety, Marcel Dekker, Inc., New York, NY.

[0362] The pharmaceutical antibody formulations can be administered by continuous infusion or at intervals such as daily, 1-7 times per week, weekly, biweekly, 3 weeks, monthly, bimonthly, etc. Preferred dosing protocols include the maximum dose or dosing frequency that avoids significant undesirable side effects. Total weekly doses are generally at least 0.05 μg / kg, 0.2 μg / kg, 0.5 μg / kg, 1 μg / kg, 10 μg / kg, 100 μg / kg, 0.2 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 10 mg / kg, 25 mg / kg, 50 mg / kg body weight or more. See, e.g., Yang et al. (2003) New Engl. J. Med. 349:427-434; Herold et al. (2002) New Engl. J. Med. 346:1692-1698; Liu et al. (1999) J. Neurol. Neurosurg. Psych. 67:451-456; Portielji et al. (20003) Cancer Immunol. Immunother. 52:133-144. Desirable dosages of small molecule therapeutics (e.g., peptidomimetics, natural products, or organic chemicals) are approximately the same as for antibodies or polypeptides, on a "moles / kg" basis.

[0363] Embodiments of the present invention further include the use of a therapeutic agent, such as a therapeutic agent for the treatment of a human body; a therapeutic agent for the treatment of a human cancer; a therapeutic agent for the treatment of a human cancer; a therapeutic agent for the treatment of a human cancer; a therapeutic agent for the treatment of a human cancer; a therapeutic agent for the treatment of a human cancer; a therapeutic agent for the treatment of a human cancer; a therapeutic agent for the treatment of a human cancer; (k) reducing the severity or duration of clinical symptoms of a PD-1 associated disease or an ILT4 associated disease; (l) prolonging patient survival compared to expected survival in a similar untreated patient; (m) inducing complete or partial remission of a cancerous condition or other PD-1 associated disease or an ILT4 associated disease; (n) treating cancer; or (o) treating a chronic infection; also encompasses one or more of the biologics described herein for use in (i), for use as a medicament or composition for (ii), or for use in the preparation of a medicament for (iii).

[0364] Common methods Standard methods in molecular biology are described in: Sambrook, Fritsch and Maniatis (1982 & 1989 2 nd Edition, 2001 3 rd Edition) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Sambrook and Russell (2001) Molecular Cloning, 3 rded., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Wu (1993) Recombinant DNA, Vol. 217, Academic Press, San Diego, CA). Standard methods are also described in Ausbel, et al. (2001) Current Protocols in Molecular Biology, Vols.1-4, John Wiley and Sons, Inc. New York, NY, which describes cloning and DNA mutagenesis in bacterial cells (Vol. 1), cloning in mammalian cells and yeast (Vol. 2), glycoconjugate and protein expression (Vol. 3), and bioinformatics (Vol. 4).

[0365] Protein purification methods, including immunoprecipitation, chromatography, electrophoresis, centrifugation and crystallization, have been described (Coligan, et al. (2000) Current Protocols in Protein Science, Vol. 1, John Wiley and Sons, Inc., New York). Chemical analysis, chemical modification, post-translational modification, production of fusion proteins, and glycosylation of proteins are described (see, e.g., Coligan, et al. (2000) Current Protocols in Protein Science, Vol. 2, John Wiley and Sons, Inc., New York; Ausubel, et al. (2001) Current Protocols in Molecular Biology, Vol. 3, John Wiley and Sons, Inc., NY, NY, pp. 16.0.5-16.22.17; Sigma-Aldrich, Co. (2001) Products for Life Science Research, St. Louis, MO; pp. 45-89; Amersham Pharmacia Biotech (2001) BioDirectory, Piscataway, NJ, pp. 384-391). The generation, purification and fragmentation of polyclonal and monoclonal antibodies have been described (Coligan, et al. (2001) Current Protocols in Immunology, Vol. 1, John Wiley and Sons, Inc., New York; Harlow and Lane (1999) Using Antibodies, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Harlow and Lane, supra).Standard techniques are available for characterizing ligand / receptor interactions (see, e.g., Coligan, et al. (2001) Current Protocols in Immunology, Vol. 4, John Wiley, Inc., New York).

[0366] Monoclonal, polyclonal and humanized antibodies can be prepared (see, e.g., Sheperd and Dean (eds.) (2000) Monoclonal Antibodies, Oxford Univ. Press, New York, NY; Kontermann and Dubel (eds.) (2001) Antibody Engineering, Springer-Verlag, New York; Harlow and Lane (1988) Antibodies A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 139-243; Carpenter, et al. (2000) J. Immunol. 165:6205; He, et al. (1998) J. Immunol. 160:1029; Tang et al. (1999) J. Biol. Chem. 274:27371-27378; Baca et al. (1997) J. Biol. Chem. 272:10678-10684; Chothia et al. (1989) Nature 342:877-883; Foote and Winter (1992) J. Mol. Biol. 224:487-499; U.S. Patent No. 6,329,511).

[0367] An alternative to humanization is to use human antibody libraries displayed on phage or in transgenic mice (Vaughan et al. (1996) Nature Biotechnol. 14:309-314; Barbas (1995) Nature Medicine 1:837-839; Mendez et al. (1997) Nature Genetics 15:146-156; Hoogenboom and Chames (2000) Immunol. Today 21:371-377; Barbas et al. (2001) Phage Display: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York; Kay et al. (1996) Phage Display of Peptides and Proteins: A Laboratory Manual, Academic Press, San Diego, CA; de Bruin et al. (1999) Nature Biotechnol. 17:397-399).

[0368] It is not necessary to purify an antigen to generate antibodies. Animals can be immunized with cells bearing the antigen of interest. Splenocytes can then be isolated from the immunized animal and fused with a myeloma cell line to produce hybridomas (see, e.g., Meyaard et al. (1997) Immunity 7:283-290; Wright et al. (2000) Immunity 13:233-242; Preston et al., supra; Kaithamana et al. (1999) J. Immunol. 163:5157-5164).

[0369] Antibodies can be conjugated, for example, to small drug molecules, enzymes, liposomes, polyethylene glycol (PEG). Antibodies are useful for therapeutic, diagnostic, kits or other purposes and include, for example, antibodies conjugated with dyes, radioisotopes, enzymes or metals, such as colloidal gold (see, for example, Le Doussal et al. (1991) J. Immunol. 146:169-175; Gibellini et al. (1998) J. Immunol. 160:3891-3898; Hsing and Bishop (1999) J. Immunol. 162:2804-2811; Everts et al. (2002) J. Immunol. 168:883-889).

[0370] Methods for flow cytometry, including fluorescence-activated cell sorting (FACS), are available (see, e.g., Owens, et al. (1994) Flow Cytometry Principles for Clinical Laboratory Practice, John Wiley and Sons, Hoboken, NJ; Givan (2001) Flow Cytometry, 2 nd ed.; Wiley-Liss, Hoboken, NJ; Shapiro (2003) Practical Flow Cytometry, John Wiley and Sons, Hoboken, NJ). Fluorescent reagents suitable for modifying nucleic acids, including nucleic acid primers and probes, polypeptides, and antibodies, for use, e.g., as diagnostic reagents, are available (Molecular Probesy (2003) Catalogue, Molecular Probes, Inc., Eugene, OR; Sigma-Aldrich (2003) Catalogue, St. Louis, MO).

[0371] Standard methods for immune system histology have been described (see, e.g., Muller-Harmelink (ed.) (1986) Human Thymus: Histopathology and Pathology, Springer Verlag, New York, NY; Hiatt, et al. (2000) Color Atlas of Histology, Lippincott, Williams, and Wilkins, Phila, PA; Louis, et al. (2002) Basic Histology: Text and Atlas, McGraw-Hill, New York, NY).

[0372] Software packages and databases are available for determining, for example, antigenic fragments, leader sequences, protein folding, functional domains, glycosylation sites, and sequence alignments (see, e.g., GenBank, Vector NTI® Suite (Informax, Inc., Bethesda, MD); GCG Wisconsin Package (Accelrys, Inc., San Diego, CA); DeCypher® (TimeLogic Corp., Crystal Bay, Nevada); Menne, et al. (2000) Bioinformatics 16: 741-742; Menne, et al. (2000) Bioinformatics Applications Note 16:741-742; Wren, et al. (2002) Comput. Methods Programs Biomed. 68:177-181; von Heijne (1983) Eur. J. Biochem. 133:17-21; von Heijne (1986) Nucleic Acids Res. 14:4683-4690).

[0373] Analysis method Suitable analytical methods for evaluating the stability of a product include size exclusion chromatography (SEC), dynamic light scattering (DLS), differential scanning calorimetry (DSC), iso-asp quantification, titer, UV at 340 nm, UV spectroscopy and FTIR. SEC (J. Pharm. Scien., 83:1645-1650, (1994); Pharm. Res., 11:485 (1994); J. Pharm. Bio. Anal., 15:1928 (1997); J. Pharm. Bio. Anal., 14:1133-1140 (1986)) measures the percentage of monomer in the product and provides information on the amount of soluble aggregates. DSC (Pharm. Res., 15:200 (1998); Pharm. Res., 9:109 (1982)) provides information on the denaturation temperature and glass transition temperature of the protein. DLS (American Lab., November (1991)) measures the average diffusion coefficient and gives information on the amount of soluble and insoluble aggregates. UV at 340 nm measures the scattered light intensity at 340 nm and gives information on the amount of soluble and insoluble aggregates. UV spectroscopy measures the absorbance at 278 nm and gives information on the protein concentration. FTIR (Eur. J. Pharm. Biopharm., 45:231 (1998); Pharm. Res., 12:1250 (1995); J. Pharm. Scien., 85:1290 (1996); J. Pharm. Scien., 87:1069 (1998)) measures the IR spectrum in the amide 1 region and gives information on the secondary structure of the protein.

[0374] The iso-asp content in the samples is measured using the Isoquant Isoaspartate Detection System (Promega). This kit uses the enzyme protein isoaspartyl methyltransferase (PIMT) to specifically detect the presence of isoaspartic acid residues in target proteins. PIMT catalyzes the transfer of a methyl group at the α-carboxyl position from S-adenosyl-L-methionine to isoaspartic acid, generating S-adenosyl-L-homocysteine ​​(SAH) in the process. This is a relatively small molecule that can usually be isolated and quantified by reversed-phase HPLC using the SAH HPLC standard that is included in the kit.

[0375] The potency or bioidentity of an antibody can be measured by its ability to bind to its antigen. Specific binding of an antibody to its antigen can be quantified by any method known to those skilled in the art, for example by an immunoassay such as ELISA (enzyme-linked immunosorbent assay).

[0376] All publications mentioned herein are incorporated by reference for the purpose of describing and disclosing methodologies and materials that might be used in connection with the present invention.

[0377] Although various embodiments of the present invention have been described herein with reference to the accompanying drawings, it is to be understood that the invention is not limited to these precise embodiments, and various changes and modifications may be made by those skilled in the art without departing from the scope or spirit of the present invention as defined in the appended claims.

[0378] Embodiment 1 comprises: (i) from about 10 mg / mL to about 200 mg / mL of an anti-human immunoglobulin-like transcript 4 ("anti-ILT4") antibody or antigen-binding fragment thereof; (ii) from about 10 mg / mL to about 200 mg / mL of an anti-PD-1 antibody or antigen-binding fragment thereof; (iii) from about 5 mM to about 20 mM of a buffer; (iv) from about 6% to about 8% weight / volume (w / v) of a non-reducing sugar; (v) from about 0.01% to about 0.10% (w / v) of a non-ionic surfactant; and and (vi) about 1 mM to about 20 mM of an antioxidant; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO:19), CDR-L2: GX1X2NRPS, where X1 is N, Q, E, or D, and X2 is S or A (SEQ ID NO:20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO:21); wherein the anti-PD-1 antibody, or antigen-binding fragment thereof, comprises a variable light chain region comprising a CDRL1 of SEQ ID NO:95, a CDRL2 of SEQ ID NO:96, and a CDRL3 of SEQ ID NO:97; and and a variable heavy chain region comprising a CDRH1 of SEQ ID NO:100, a CDRH2 of SEQ ID NO:101, and a CDRH3 of SEQ ID NO:102; wherein the buffer is an L-histidine buffer, an acetate buffer, or a citrate buffer, wherein the non-reducing sugar is a disaccharide, wherein the non-ionic surfactant is polysorbate 20 or polysorbate 80, and wherein the antioxidant is methionine or a pharma- ceutically acceptable salt thereof.

[0379] Embodiment 2 provides the formulation of embodiment 1, comprising 40-60 mg / mL of the anti-ILT4 antibody; 15-35 mg / mL of the anti-PD-1 antibody; about 7% sucrose or trehalose; about 0.025% polysorbate 80 or polysorbate 20; about 3-30 mM L-histidine buffer having a pH of about 5.0-6.5; and about 5-20 mM methionine or a pharma-ceutically acceptable salt thereof.

[0380] Embodiment 3 provides the formulation of embodiment 1, comprising 45-55 mg / mL of the anti-ILT4 antibody; 20-30 mg / mL of the anti-PD-1 antibody; about 7% sucrose or trehalose; about 0.025% polysorbate 80 or polysorbate 20; about 3-30 mM L-histidine buffer having a pH of about 5.0-6.5; and about 10-20 mM L-methionine or a pharma-ceutically acceptable salt thereof.

[0381] Embodiment 4 provides the formulation of embodiment 1, wherein the buffer is an L-histidine buffer.

[0382] Embodiment 5 provides a formulation of any one of embodiments 1-4, wherein the non-reducing sugar is sucrose.

[0383] Embodiment 6 provides a formulation of any one of embodiments 1-5, wherein the non-ionic surfactant is polysorbate 80.

[0384] Embodiment 7 provides a formulation of any one of embodiments 1-6, wherein the antioxidant is L-methionine.

[0385] Embodiment 8 provides a formulation comprising: (i) about 10 mg / mL to about 200 mg / mL of an anti-ILT4 antibody ("anti-ILT4") or antigen-binding fragment thereof; (ii) about 10 mg / mL to about 200 mg / mL of an anti-PD-1 antibody or antigen-binding fragment thereof; (iii) about 5 mM to about 20 mM L-histidine buffer; (iv) about 6% to about 8% weight / volume (w / v) sucrose; (v) about 0.01% to about 0.10% (w / v) polysorbate 80; and (vi) about 1 mM to about 20 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof has the structure: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17), and CDR-H3: and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO:19), CDR-L2: GX1X2NRPS, where X1 is N, Q, E, or D, and X2 is S or A (SEQ ID NO:20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO:21); and wherein the anti-PD-1 antibody, or antigen-binding fragment thereof, comprises a variable light chain region comprising a CDRL1 of SEQ ID NO:95, a CDRL2 of SEQ ID NO:96, and a CDRL3 of SEQ ID NO:97; and a variable heavy chain region comprising a CDRH1 of SEQ ID NO:100, a CDRH2 of SEQ ID NO:101, and a CDRH3 of SEQ ID NO:102; or a pharma- ceutically acceptable salt thereof.

[0386] Embodiment 9 provides the formulation of embodiment 8, comprising about 8 mM to about 12 mM L-histidine buffer.

[0387] Embodiment 10 provides a formulation of any one of embodiments 8 or 9, comprising about 5 mM to about 10 mM L-methionine.

[0388] Embodiment 11 provides a formulation according to any one of embodiments 8 to 10, comprising about 0.01% to about 0.05% (w / v) polysorbate 80.

[0389] Embodiment 12 provides the formulation of any one of Embodiments 1 or 4-11, comprising about 10 mg / mL to about 150 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof.

[0390] Embodiment 13 provides a formulation of any one of Embodiments 1 or 4-11, comprising about 10 mg / mL to about 150 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof.

[0391] Embodiment 14 provides the formulation of any one of Embodiment 12, wherein the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 10 mg / mL, about 12.5 mg / mL, about 25 mg / mL, about 50 mg / mL, about 75 mg / mL, about 100 mg / mL, about 125 mg / mL or about 150 mg / mL.

[0392] Embodiment 15 provides the formulation of any one of Embodiment 12, wherein the concentration of the anti-PD-1 antibody or antigen-binding fragment thereof is about 10 mg / mL, about 12.5 mg / mL, about 25 mg / mL, about 50 mg / mL, about 75 mg / mL, about 100 mg / mL, about 125 mg / mL, or about 150 mg / mL.

[0393] Embodiment 16 provides a formulation of any one of Embodiments 1-13, comprising about 25 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

[0394] Embodiment 17 provides a formulation of any one of Embodiments 1-13, comprising about 50 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 25 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

[0395] Embodiment 18 provides a formulation of any one of Embodiments 1-13, comprising about 75 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 50 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

[0396] Embodiment 19 provides a formulation of any one of Embodiments 1-13, comprising about 100 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 75 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

[0397] Embodiment 20 provides a formulation of any one of Embodiments 1-13, comprising about 125 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 100 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

[0398] Embodiment 21 provides a formulation according to any one of embodiments 1 to 20, wherein the formulation has a pH of about 5.0 to about 6.8.

[0399] Embodiment 22 provides a formulation according to any one of embodiments 1 to 21, wherein the formulation has a pH of about 5.5 to about 6.0.

[0400] Embodiment 23 provides a formulation of any one of embodiments 1 to 22, wherein the formulation has a pH of about 5.5.

[0401] Embodiment 24 is a formulation of an anti-human immunoglobulin-like transcript 4 ("anti-ILT4") antibody or antigen-binding fragment thereof comprising: (i) about 50 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mg / mL to about 200 mg / mL of an anti-PD-1 antibody or antigen-binding fragment thereof; (iii) about 10 mM L-histidine buffer, pH about 5.5; (iv) about 7% weight / volume (w / v) sucrose; (v) about 0.025% (w / v) polysorbate 80; and (vi) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof has the structure of CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO:19), CDR-L2: GX1X2NRPS, where X1 is N, Q, E, or D, and X2 is S or A (SEQ ID NO:20) and CDR-L3: QSFDNSLSAYV (SEQ ID NO:21); and wherein the anti-PD-1 antibody, or antigen-binding fragment thereof, comprises a variable light chain region comprising a CDRL1 of SEQ ID NO:95, a CDRL2 of SEQ ID NO:96, and a CDRL3 of SEQ ID NO:97; and a variable heavy chain region comprising a CDRH1 of SEQ ID NO:100, a CDRH2 of SEQ ID NO:101, and a CDRH3 of SEQ ID NO:102.

[0402] Embodiment 25 provides the formulation of any one of embodiments 1 to 24, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E, or D and X2 is S or A) (SEQ ID NO: 20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0403] Embodiment 26 provides the formulation of any one of Embodiments 1 to 24, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0404] Embodiment 27 provides the formulation of any one of Embodiments 1 to 24, wherein the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:58.

[0405] Embodiment 28 provides the formulation of any one of Embodiments 1 to 24, wherein the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2; and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0406] Embodiment 29 provides the formulation of any one of Embodiments 1 to 24, wherein the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:80; and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0407] Embodiment 30 provides the formulation of any one of Embodiments 1 to 24, wherein the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO: 57; and a light chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO: 58.

[0408] Embodiment 31 provides the formulation of any one of Embodiments 1 to 24, wherein the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:2; and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7.

[0409] Embodiment 32 provides the formulation of any one of Embodiments 1 to 24, wherein the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 80; and a light chain consisting of the amino acid sequence set forth in SEQ ID NO: 7.

[0410] Embodiment 33 provides the formulation of any one of Embodiments 1 to 24, wherein the anti-human PD-1 antibody, or antigen-binding fragment thereof, comprises a variable light chain region comprising the amino acid sequence set forth in SEQ ID NO: 98; and a variable heavy chain region comprising the amino acid sequence set forth in SEQ ID NO: 103.

[0411] Embodiment 34 provides the formulation of any one of Embodiments 1 to 24, wherein the anti-human PD-1 antibody, or antigen-binding fragment thereof, comprises a variable light chain region comprising the amino acid sequence set forth in SEQ ID NO: 99; and a variable heavy chain region comprising the amino acid sequence set forth in SEQ ID NO: 104.

[0412] Embodiment 35 provides the formulation of any one of Embodiments 1 to 24, wherein the anti-human PD-1 antibody, or antigen-binding fragment thereof, comprises two light chains and two heavy chains, wherein the two light chains comprise the amino acid sequence set forth in SEQ ID NO: 99, and the two heavy chains comprise the amino acid sequence set forth in any one of SEQ ID NOs: 104-109.

[0413] Embodiment 37 provides the formulation of any one of embodiments 1 to 35, wherein the anti-human PD-1 antibody is pembrolizumab.

[0414] Embodiment 37 provides a formulation of any one of Embodiments 1 to 36, wherein the ratio of anti-ILT4 antibody to anti-PD-1 antibody is 1:1 or 1:2.

[0415] Embodiment 38 provides a formulation according to any one of embodiments 1 to 37, wherein after 9 months at 5° C., (i) the % monomer of the anti-ILT4 antibody is ≧95% when measured by size exclusion chromatography; (ii) the % heavy chain and % light chain of the anti-ILT4 antibody are ≧90% when measured by reduced CE-SDS; (iii) the % heavy chain and % light chain of the anti-ILT4 antibody are ≧95% when measured by reduced CE-SDS; (iv) the % intact IgG of the anti-ILT4 antibody is ≧90% when measured by non-reduced CE-SDS; and / or (v) the % intact IgG of the anti-ILT4 antibody is ≧95% when measured by non-reduced CE-SDS.

[0416] Embodiment 39 provides the formulation of any one of embodiments 1 to 38, wherein the formulation is a liquid formulation stored at about 3° C. to about 5° C. or a reconstituted solution from a lyophilized formulation.

[0417] Embodiment 40 is a method for producing a composition comprising the steps of: (i) storing said anti-ILT4 antibody or antigen-binding fragment thereof at about 3° C. to about 5° C. for up to 9 months; (ii) storing said anti-ILT4 antibody or antigen-binding fragment thereof at about 3° C. to about 5° C. for up to 9 months; and (iii) storing said anti-ILT4 antibody or antigen-binding fragment thereof at about 3° C. to about 5° C. for up to 9 months; (i) the % monomer of said anti-ILT4 antibody or antigen-binding fragment thereof is at least about 99% as measured by ultra-performance size exclusion chromatography; and (ii) the OD 350-500(iii) the turbidity of the formulation is at most about 0.135 when measured by high performance ion exchange chromatography; (iv) the number of subvisible particles of particles at least 2□m in size is at most about 3500 when measured by microflow imaging; and / or (v) the % oxidation of one or more amino acid residues selected from the group consisting of W7, W102, M253, M359 and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% when measured by reduced peptide mapping;

[0418] Embodiment 41 provides the formulation of any of Embodiments 1-34 or 37-41, wherein the anti-PD-1 antibody, or antigen-binding fragment thereof, is a monoclonal antibody.

[0419] Embodiment 42 provides a method of treating cancer in a human patient in need thereof, comprising administering an effective amount of the formulation of any one of embodiments 1-40.

[0420] Embodiment 43 provides the method of embodiment 42, wherein the cancer is selected from the group consisting of colorectal cancer, esophageal cancer, melanoma, non-small cell lung cancer, ovarian cancer, renal cell carcinoma, and small cell lung cancer.

[0421] Embodiment 44 provides the use of a formulation according to any one of embodiments 1 to 41 for the preparation of a medicament for treating cancer.

[0422] Embodiment 45 provides the use of a formulation according to any one of embodiments 1 to 42 for treating cancer in a human patient.

[0423] Embodiment 46 provides the use according to embodiment 45, wherein the cancer is selected from the group consisting of colorectal cancer, esophageal cancer, melanoma, non-small cell lung cancer, ovarian cancer, renal cell carcinoma and small cell lung cancer. EXAMPLES

[0424] Working Example Example 1: Preparation of anti-ILT4 / anti-PD-1 co-formulations Anti-ILT4 monoclonal antibody and anti-PD-1 antibody ("anti-ILT4 / anti-PD-1 co-formulation") injection was prepared as a sterile, preservative-free solution requiring dilution for intravenous infusion. Anti-ILT4 / anti-PD-1 co-formulation is a fixed-dose combination of anti-ILT4 antibody and pembrolizumab antibody. The co-formulation composition was manufactured by combining fully formulated anti-ILT4 monoclonal antibody Process 2 drug substance and pembrolizumab drug substance. The anti-ILT4 drug substance contained 50.0 mg / mL anti-ILT4 in 0.29 mg / mL L-histidine, 1.71 mg / mL L-histidine monohydrochloride monohydrate, 1.49 mg / mL L-methionine, 70.0 mg / mL sucrose, and 0.25 mg / mL polysorbate 80 (pH 5.5). Pembrolizumab drug substance contained 25.0 mg / mL pembrolizumab in 0.29 mg / mL L-histidine, 1.71 mg / mL L-histidine monohydrochloride monohydrate, 70 mg / mL sucrose, and 0.20 mg / mL polysorbate 80 (pH 5.5). The following formulation attributes were considered during formulation optimization: protein concentration, buffer, pH, charged species, antioxidants, and surfactants.

[0425] Example 2: Pre-anti-ILT4 / Pembrolizumab Co-Formulation Analysis The pre-co-formulation development plan is summarized in Table 6. The studies were conducted to evaluate the sensitivity of the anti-ILT4 drug product (DP), single entity (SE) and co-formulation with pembrolizumab (CF) to various formulation and process parameters that the products experience during manufacturing and are previously known to affect oxidation, such as light exposure, oxygen, and residual levels of peroxides and metals. Filtered bulk DP of anti-ILT4 (30 mg / mL), pembrolizumab (7.5 mg / mL), and anti-ILT4 + pembrolizumab (30 mg / mL + 7.5 mg / mL) were spiked with residual levels of peroxide (0, 20, and 200 ppb), metals (0, 25, and 250 ppb), and exposed to light stress (0x, 0.2x, and 0.4x ICH of a combination of CWL + UV (where 1x ICH exposure level was 1.2 million lux hours of white light and 200 watt-hours / m2). 2 The anti-ILT4 was subjected to 1000 s UV energy (corresponding to 1000 s UV energy) and then stability staged for up to 12 weeks at 5°C and 4 weeks at 40°C to assess potential oxidative liability of the anti-ILT4. The range of stress levels tested for metal, peroxide and light exposure were selected to cover the target exposures during DP production at a commercial manufacturing site and were then appropriately bracketed with respect to control and worst case scenarios. Additionally, this study also served to set the free L-methionine concentration (7, 10 and 15 mM) as an excipient to inhibit protein oxidation. Samples were analyzed by HIC, IEX, SEC, L-methionine content, PS-80 content, subvisible particles and reduced peptide mapping assays.

[0426] The compositions of the anti-ILT4 / anti-PD-1 co-formulations tested in this study are shown in Table 6. L-methionine concentrations (7, 10, and 15 mM) were set as an excipient to control protein oxidation.

[0427] [Table 6]

[0428] Example 3: Evaluation of oxidation sensitivity The five key parameters examined in this study were L-Met, peroxide, metals, fill volume, and photostress. The program approved a D-optimal split-plot design of 36 formulations each (108 formulations total) for anti-ILT4 / anti-PD-1 coformulations, anti-ILT4 mono entity (SE), and pembrolizumab SE. Since photostress is a difficult factor to change, the 36 formulations (data not shown) had six whole plots, and the photostress level was constant for each of the whole plots. Each whole plot was further divided into two subplots to ensure that L-Met levels were the same within a plot. The ranges investigated for the five factors are shown in Table 7.

[0429] [Table 7]

[0430] Example 4: Preparation of vials with added metal and peroxide Samples were prepared using diluents (consisting of protein-free solutions) of the same composition as the stock solutions of anti-ILT4 / anti-PD-1 co-formulations, anti-ILT4 SE, and pembrolizumab SE. Metal stock solutions were prepared using an equal volume of FeCl 2 and CuCl 2 Specifically, the 100 ppb metal stock solution contained 50 ppb FeCl 2 and 50 ppb of CuCl 2The peroxide stock solutions contained equal amounts of t-BHP and AAPH. Specifically, the 100 ppb peroxide stock solution contained 50 ppb t-BHP and 50 ppb AAPH. Using the diluent, peroxide stock solution, and metal stock solutions, appropriate amounts of these solutions were added to the filtered anti-ILT4 / anti-PD-1 co-formulations, anti-ILT4 SE, and pembrolizumab SE stock formulations in a ratio of 1 μL of diluent, peroxide stock solution, or metal stock solution for each 1 mL of stock formulation used. As such, multiple combinations of peroxide and metal concentrations were targeted. Specifically, the peroxide concentrations evaluated were 0, 20, and 200 ppb, while the metal concentrations evaluated were 0, 25, and 250 ppb. Considering different L-methionine concentrations (7, 10, and 15 mM), loading volumes (1.2, 2.2, and 3.2 mL) and light exposure levels (0x, 0.2x, and 0.4x ICH CWL+UV), a total of 108 independent formulations were prepared for anti-ILT4 / anti-PD-1 co-formulations, anti-ILT4 SE, and pembrolizumab SE. Table 7 summarizes the oxidative stress conditions for each of the 108 formulations prepared and the volumes of stock solutions used to prepare them. In this study, PS-80 levels were kept constant in all formulations and degradation was monitored over the study parameters and study period.

[0431] The filtered anti-ILT4 / anti-PD-1 co-formulations, anti-ILT4 SE and anti-ILT4 SE formulations were filled into 2R vials at various fill volumes (1.2, 2.2 and 3.2 mL) in a laminar flow hood. The different fill volumes used in this study were chosen to control the amount of oxygen available in the vial. The 2R vials were used in place of the intended commercial images as scale-down models and to minimize material usage (20 mL and 30 mL vials for the mono and co-formulation images, respectively).

[0432] The same 36 formulations (data not shown) were prepared for anti-ILT4 / anti-PD-1 CF, anti-ILT4 SE, and pembrolizumab SE. Stock formulations corresponded to the corresponding co-formulation or monoformulation stocks and depended on the L-methionine concentration (7, 10, or 15 mM).

[0433] Example 5: Light Exposure and Stability Studies All vials of the same formulation set (i.e., anti-ILT4 / anti-PD-1 co-formulation, anti-ILT4 SE, or pembrolizumab SE) were placed in a photostability chamber at 25°C for light exposure (0x, 0.2x, and 0.4x ICH CWL+UV). Samples intended for 0x ICH light exposure were protected from light, but these samples were also left in the chamber for the duration of the experiment. Samples for 0.2x and 0.4x ICH were placed horizontally in the chamber to maximize exposure.

[0434] After all vials were exposed to light, samples were stability staged at 5° C. for up to 12 weeks and at 40° C. for up to 4 weeks according to the protocol in Table 2. At each stability time point, samples were drawn and frozen at −80° C. before being submitted for analytical testing (with appropriate controls).

[0435] Example 6: Qualitative Analysis Visual analysis of the analytical data suggested no effect of increasing residual peroxide levels (0 → 200 ppb) or changing fill volume (1.2 → 3.2 mL). On the other hand, changes in light exposure (0x → 0.4x ICH) and residual metal content (0 → 250 ppb) increase drug product degradation as observed by changes in high molecular weight species in UP-SEC (Figure 1), changes in degradation of PS80 (Figure 2), and changes in oxidation of methionine and tryptophan residues from reduced peptide mapping (Figures 3, 4, and 5) and HIC (Figure 6). It is noted that the observed increase in physical and chemical degradation was detected in samples stored at 40°C for up to 4 weeks, whereas no changes were observed in samples stored at 5°C for up to 3 months. Results for other attributes such as charge variants (by IEX) and sub-visible particles (by MFI) show little or no change with varying peroxide levels, metal levels, light levels or fill volume levels, as shown in the data summary.

[0436] Collectively, the data from this study showed that light exposure and metal stress were the most important factors affecting the oxidation of anti-ILT4 / anti-PD-1 co-formulations. Nevertheless, the impact from these stress factors was reduced by increasing the L-methionine concentration. As such, the data suggested that a concentration of 15 mM L-methionine resulted in the lowest levels of aggregation and chemical degradation. Of note, when the L-methionine concentration was higher than 10 mM, no improvement in the physical and chemical properties of anti-ILT4 monoclonal antibody SE and pembrolizumab SE was observed, so such L-methionine concentrations were only effective for anti-ILT4 / anti-PD-1 co-formulations.

[0437] Example 7: UP-SEC (Ultra Performance Size Exclusion Chromatography) The increase in high molecular weight species (HMW) and low molecular weight species (LMW) indicates protein degradation via aggregation and fragmentation. The plots (data not shown) showed that higher light stress levels resulted in higher %HMW and %LMW, which increased with time, while the 40°C samples showed a lower percentage of monomer, which further decreased with time.

[0438] Statistical analysis of the data was performed to evaluate the impact of various formulation and process factors (i.e., L-methionine concentration, peroxide content, metal content, light exposure, and loading amount). Based on this analysis, contour plots of %HMW only were provided for all three formulations.

[0439] Figure 7 shows a contour plot for the HMW co-formulation with predicted values ​​from model fits at various settings of three key parameters (L-methionine, light exposure, and metal content) over time at 40°C. The contour plot shows that the predicted levels of HMW increase with increasing light stress and with lower levels of L-Met.

[0440] Similar to the co-formulations, the contour plots for anti-ILT4 monoclonal antibody SE shown in FIG. 8 show that when L-methionine concentrations are below 10 mM, HMW levels increase with increasing light exposure and metal content.

[0441] A contour plot for pembrolizumab SE is shown in Figure 9. As with anti-ILT4SE, statistical analysis showed that HMW levels increased with increasing light exposure and metal content when L-methionine concentrations were below 10 mM.

[0442] The changes in charge variants are likely the result of oxidation, deamidation, and isomerization. Mutability plots (data not shown) showed that higher light stress levels increased % acidic and % basic variants and decreased % major components under all conditions. The same conclusion was seen for pembrolizumab charge variants (both coformulation and pembrolizumab SE).

[0443] Statistical analysis of the data was performed to evaluate the influence of various formulation and process factors (i.e., L-methionine concentration, peroxide content, metal content, light exposure, and loading amount). Based on this analysis, contour plots of % acidic variants, % major components, and % basic variants are presented for all three formulations in Figures 10-18.

[0444] The contour plot in FIG. 10 shows the predicted % acidic variants for anti-ILT4 in co-formulations at various factor levels at 40° C. for 4 weeks.

[0445] The contour plot in Figure 11 shows the % major component for anti-ILT4 in the co-formulation as a function of various factors (L-methionine concentration, light exposure, and metal content). The figure illustrates that the % major component for anti-ILT4 decreases with increasing light stress.

[0446] The contour plot in Figure 12 shows the % basic variants for anti-ILT4 in the co-formulation as a function of various factors (L-methionine concentration, light exposure, and metal content). The figure shows that increasing light stress increases the % basic variants for anti-ILT4.

[0447] The contour plot in Figure 13 shows the predicted values ​​from the model fit at various settings of the three parameters (L-methionine concentration, light exposure, and metal content) over time. The contour plot shows that as light stress increases, the predicted levels of % acidic variants for pembrolizumab in the co-formulation remain constant.

[0448] The contour plot in Figure 14 shows the % major component for pembrolizumab in the co-formulation as a function of various factors (L-methionine concentration, light exposure, and metal content). The figure shows that the % major component for pembrolizumab decreases with increasing light stress and metal content. In contrast, increasing the L-methionine concentration above 10 mM reduces the rate of decrease in major component due to light stress.

[0449] The contour plot in Figure 15 shows the % basic variants for pembrolizumab in the co-formulation as a function of various factors (L-methionine concentration, light exposure, and metal content). The figure shows that increasing light stress decreases the % basic variants for pembrolizumab, while increasing the L-methionine concentration above 10 mM reduces the rate of loss of the major components due to light stress.

[0450] The contour plot in Figure 16 shows the predicted values ​​from the model fit at various settings of the three parameters (L-methionine concentration, light exposure, and metal content) over time. The contour plot shows that as light stress increases, the predicted levels of % acidic variants for anti-ILT4 in the anti-ILT4 SE formulation increase.

[0451] The contour plot in Figure 17 shows the % major component for anti-ILT4 in the anti-ILT4 SE formulation as a function of various factors (L-methionine concentration, light exposure, and metal content). The figure shows that increasing light stress and metal content decrease the % major component for anti-ILT4.

[0452] The contour plot in Figure 18 shows the % basic variants for anti-ILT4 in the anti-ILT4 SE formulation as a function of various factors (L-methionine concentration, light exposure, and metal content). The figure shows that with increasing light stress, the % basic variants for anti-ILT4 increases.

[0453] Overall, the data provided important information on the five key parameters used to study the oxidation process. In some of the responses, outlying values ​​introduced noise into the data, making statistical interpretation difficult. However, a comprehensive review of the data based on statistical models, predictive profilers, and contour plots suggested final formulations for the co-formulation and anti-ILT4 SE formulations. Based on the response of attributes measuring oxidation, high levels of light stress consistently had a significant effect on all reactions. L-methionine concentration, peroxide content, and metal content were also significant for some of the reactions. Metal content showed a significant effect on the pembrolizumab SE formulation in some cases. Fill weight did not have a significant effect on most of the reactions, and the factor range could be kept at the proposed level of 1.2 mL to 3.2 mL. For prediction purposes, the fill volume was kept at a target level of 2.2 mL.

[0454] Based on statistical predictions of key responses (UPSEC, IEX, PS-80, L-methionine, turbidity), data summaries, graphs and scientific inputs, the final formulations selected for SE and co-formulated products are shown in Table 8.

[0455] [Table 8]

[0456] In this study, the long-term stability of two lead formulation prototypes was evaluated. The final two compositions for the anti-ILT4 / anti-PD-1 co-formulations (containing 10 mM and 15 mM L-methionine, respectively) were run for stability at 5°C for 12 months and at 25°C and 40°C for 6 months. The formulation compositions for co-formulation 1 (CF1, 15 mM L-methionine) and co-formulation 2 (CF2, 10 mM L-methionine) are shown in Table 9. All data are shown in Tables 10-12. Excess 5 mM L-methionine was added as a powder and dissolved by gentle mixing. Samples were analyzed by HIC, IEX, SEC, L-methionine content, PS-80 content, subvisible particles, and reduced peptide mapping assays.

[0457] L-Methionine concentrations at all stability time points for all three formulations as shown in Figure 19. The results indicate that the effect of all different factors on the stability of L-Methionine is negligible.

[0458] [Table 9]

[0459] [Table 10] TIFF2025514174000013.tif30169

[0460] [Table 11] TIFF2025514174000015.tif21168

[0461] [Table 12] TIFF2025514174000017.tif25169

[0462] Example 8: Pre-probe stability studies on anti-ILT4 / anti-PD-1 co-formulations As shown in Figure 20, a plot of turbidity values ​​for all three formulations under conditions of 5°C and 40°C. A statistical analysis of the data in Figure 20 was performed to evaluate the influence of various formulation and process factors (i.e., L-methionine concentration, peroxide content, metal content, light exposure, and fill volume). Based on this analysis, a predictive profiler of turbidity was presented for all three formulations. Increased solution turbidity indicates increased aggregation in the protein formulation.

[0463] Figures 21-25 show peptide mapping (PTM) for mono- and co-formulations under initial and 40°C conditions. Data on the main response to PTM was collected for a subset of conditions at initial and 4 weeks at 40°C for the co-formulations and anti-ILT4 SE formulations. A statistical model with main effects and a two-way interaction of the two stability time points at 40°C was fitted for all PTM responses. Reduced peptide mapping was used to monitor M105 oxidation for anti-PD-1 and multiple oxidation hotspots for the anti-ILT4 monoclonal antibody within the anti-ILT4 / anti-PD-1 co-formulation. The PTM data support the final formulations selected for the co-formulations and anti-ILT4 SE formulations. Specifically, FIG. 21 shows M253 (shared), FIG. 22 shows M429 (shared), FIG. 23 shows W7 (anti-ILT4), FIG. 24 shows W102 (anti-ILT4), and FIG. 25 shows M105 (pembrolizumab).

[0464] M105 oxidation for the anti-PD-1 monoclonal antibody component is a known issue from experience with other products co-formulated with pembrolizumab (anti-PD-1 monoclonal antibody), and the impact of increasing L-methionine (15 mM in CF1) on stability was evaluated by monitoring the change in the M105 residue. Figure 25 shows the increasing trend for all residues for both CF1 and CF2 over 6 months of stability at 40°C. CF1 (15 mM L-methionine) showed a slight reduction in M105 oxidation at 3 and 6 months at 40°C compared to CF2 (10 mM L-methionine). A slight increase in the M105 residue was observed after 6 months at 25°C and 12 months at 5°C, but the slope of increase was low, indicating minimal impact on stability at the recommended 5°C storage conditions. A similar stability trend was observed for oxidation of W102 and W7 residues of anti-ILT4 monoclonal antibodies in CF1 and CF2, where oxidation increased after 6 months at 40°C and no measurable change was observed after 12 months at the recommended storage conditions at 5°C.

[0465] Soluble aggregates (UP-SEC) High molecular weight (HMW) and low molecular weight (LMW) species measured by UP-SEC for co-formulation 1 (CF1) and co-formulation 2 (CF2) when stored for 12 months at 5° C. (circles), 6 months at 25° C. (upward triangles), and 6 months at 40° C. (downward triangles), as shown in FIG. 26. The lines correspond to third order polynomial fits and are used in the figures only as guidance. Both formulations showed a trend of increasing HMW upon storage stability at 40° C. over 6 months. There was a slight increase in HMW at 25° C. over 6 months. Aggregate content did not change up to 12 months at 5° C., indicating sufficient physical stability under the recommended storage conditions. No measurable differences in soluble aggregates were observed between co-formulation prototypes with L-methionine content varying from 10 mM to 15 mM.

[0466] Charge variants (HP-IEX) Charge variants of anti-ILT4 and pembrolizumab measured by IEX in CF1 and CF2 when stored for 12 months at 5° C. (circles), 6 months at 25° C. (upward triangles), and 6 months at 40° C. (downward triangles), as shown in Figure 27. The lines correspond to third order polynomial fits and are used in the figure only as guidance.

[0467] CF1 and CF2 were evaluated for chemical degradation by IEX-Combination method, where two monoclonals (anti-ILT4 monoclonal antibody and pembrolizumab) were resolved into a main peak and acidic / basic charged variants. Both formulations showed a stability-based decrease in the main peak and an increase in acidic and basic variants at 40°C. A slight decrease in the main peak and an increase in charged variants was observed at 25°C for 6 months, and very little change in charged species was observed at 5°C for 12 months. No measurable difference in chemical degradation was observed in the co-formulation prototype with increased L-methionine content up to 15 mM.

[0468] As shown in FIG. 28, total L-methionine content in CF1 and CF2 when stored for 12 months at 5° C. (circle symbol), 6 months at 25° C. (downward triangle symbol), and 6 months at 40° C. (upward triangle symbol).

[0469] As shown in Figure 29, total PS-80 content in CF1 and CF2 after 12 months of storage at 5°C (circles), 6 months at 25°C (triangles pointing up) and 6 months at 40°C (triangles pointing down). The lines correspond to third order polynomial fits and are used in the figure only as guidance. A temperature-dependent decrease in PS-80 was observed after 6 months of storage at 25°C and 40°C. Within the variability of the analytical assay, no change was observed in the PS-80 content of CF1 and CF2 after 12 months of storage at 5°C.

[0470] Figure 30 shows the number of subvisible particles in CF1 and CF2 when stored for 12 months at 5°C (light grey), 6 months at 25°C (dark grey) and 6 months at 40°C (black). A temperature-dependent increase in particle number versus stability was observed in both CF1 and CF2 formulations. Overall, particle numbers for both small particles (>2μm and >5μm) and large particles (>10μm and >25μm) remained low upon stability.

[0471] Figure 31 shows the oxidation of M105 (anti-PD-1), M429 (shared residue) and M253 (shared residue) as measured by reduced peptide mapping of CF1 and CF2 upon storage for 12 months at 5° C. (circles), 6 months at 25° C. (up triangles) and 6 months at 40° C. (down triangles). The lines correspond to third order polynomial fits and are used in the figure for guidance only.

[0472] Figure 32 shows the oxidation of W102 (an anti-ILT4 monoclonal antibody) and W7 (an anti-ILT4 monoclonal antibody) as measured by reduced peptide mapping of CF1 and CF2 upon storage for 12 months at 5° C. (circles), 6 months at 25° C. (upward triangles) and 6 months at 40° C. (downward triangles). The lines correspond to third order polynomial fits and are used in the figure only as guidance.

[0473] Formulation studies with fixed dose ratios of anti-ILT4 monoclonal antibody and anti-PD-1 monoclonal antibody demonstrated the feasibility of developing an anti-ILT4 / anti-PD-1 co-formulation to accommodate a fixed dose of 800 mg anti-ILT4 monoclonal antibody and 200 mg anti-PD-1 monoclonal antibody.

[0474] Based on the experimental design and pre-probe stability studies on the coformulations, the following conclusions were made: * The anti-ILT4 / anti-PD-1 co-formulation has a pH of 5.5, which has been found to be optimal for stability; *L-histidine buffer (10 mM) was identified as an acceptable buffer system for anti-ILT4 / anti-PD-1 co-formulation; *10 mM L-histidine (pH 5.5) matrix as a heat stabilizer was supplemented with sucrose at a concentration of 70 mg / mL; *PS80 at 0.25 mg / mL was chosen as the surfactant selected based on its ability to improve stability against agitation stress; *L-methionine was added at 2.24 mg / mL (15 mM) as an antioxidant to minimize oxidation under light stress.

[0475] In a formulation feasibility (Design of Experiments) study, the thermal stability of the anti-ILT4 / anti-PD-1 co-formulation was evaluated at 5°C for 12 weeks and at 40°C for 4 weeks after subjecting the samples to various stress conditions (process and formulation factors).

[0476] Example 9: Manufacturing and DP Image Considerations Diluent Selection: For anti-ILT4 / anti-PD-1 co-formulations, process diluents are added in the drug product manufacturing process to maintain the recommended levels of L-methionine (15 mM or 2.24 mg / mL) and PS-80 (0.25 mg / mL) in the final composition. Based on the extent of previous co-formulation manufacturing experience at the DP manufacturing facility (Carlow), the optimal composition was selected as a process diluent containing 0.29 mg / mL L-histidine, 1.71 mg / mL L-histidine monohydrochloride monohydrate, 17.46 mg / mL L-methionine, 70 mg / mL sucrose, and 0.45 mg / mL polysorbate 80 in water for injection. To maintain the drug product ratio and accommodate the diluent in the appropriate fill volume, the final ratio was optimized as 8:1:4 (v:v:v) anti-ILT4 monoclonal antibody:diluent:anti-PD-1 monoclonal antibody.

[0477] Filling volume: The final fill volume was optimized based on the following considerations: 1. Dilute to ensure the 8:1:4 ratio of anti-ILT4:diluent:anti-PD-1 is maintained in the final DP image; 2. The retrievable fill volume should be rounded to the nearest appropriate decimal point to allow healthcare professionals to accurately measure the volume of DP using syringe calibration (e.g., if a 30 mL syringe is used for 26.7 mL, 26.00 mL is the preferred withdrawal volume).

[0478] The final composition (30.8 mg / mL anti-ILT4 monoclonal antibody, 7.7 mg / mL anti-PD-1 monoclonal antibody, 0.29 mg / mL L-histidine, 1.71 mg / mL L-histidine monohydrochloride monohydrate, 2.24 mg / mL L-methionine, 70 mg / mL sucrose, 0.25 mg / mL polysorbate 80, pH 5.5) was found to be stable under refrigerated conditions (Table 12).

[0479] The stability of the final drug product is evaluated under storage conditions of 5°C ± 3°C, accelerated storage conditions of 25°C / 60% RH terminated after 6 months, and stress conditions of 40°C / 75% RH terminated after 6 months.

[0480] Tables 13-15 below summarize the stability data for the co-formulations. For the final drug product, no discernible changes were observed in any of the assays for stability at the recommended storage conditions (5°C). Test results for visible particles under all conditions evaluated to date are reported as "liquid essentially free of visible particles," which meets the acceptance criteria. Observations do not change as a function of storage condition or time. Potency stability data for storage conditions of 5°C, 25°C, and 40°C are shown in Tables 13-15. At the accelerated condition of 40°C, a decrease is observed over the first six months. At the stressed condition of 25°C and the recommended condition of 5°C, only slight changes in potency are observed. For the anti-PD-1 monoclonal antibody component, a slight increase in concentration is observed at the stressed condition of 6M. No other changes in protein concentration were observed at all time points and conditions.

[0481] [Table 13]

[0482] Studies were conducted to justify the amount of PS-80 in the final composition to protect against freeze-thaw (F / T) and agitation induced particle formation and aggregation. Freeze-thaw (F / T) and agitation stress studies were conducted on selected formulation compositions as shown in Table 14. Two F / T excursion conditions were studied as shown below: (a) a temperature excursion study consisting of up to 5 cycles of temperature exposure alternating between -40°C and 25°C, and (b) a temperature excursion study consisting of up to 5 cycles of temperature exposure alternating between -80°C and 25°C.

[0483] [Table 14]

[0484] Agitation stress studies under ambient conditions in which pharmaceutical vials were agitated at a speed of 300 rpm for 3 and 7 days as shown in Table 15. Pharmaceutical vials also under ambient conditions but not subjected to agitation served as controls.

[0485] [Table 15]

[0486] No changes were observed in turbidity by MFI, high molecular weight species, charge variants, and subvisible particle counts upon exposure to freeze-thaw between -40°C and 25°C for up to five freeze-thaw cycles (5X F / T). A decrease in PS-80 and L-methionine content was observed at 5X F / T for both -40°C to 25°C and -80°C to 25°C. Results are shown in Table 20.

[0487] No changes were observed in turbidity, high molecular weight species, charge variants, L-methionine content and PS-80 content up to 7 days under ambient temperature conditions. A slight increase in the number of sub-visible particles was observed for particles >= 2 μm and >= 5 μm in the MFI results for drug product stirred for 7 days compared to the 7 day control. The results are shown in Table 21.

[0488] Data obtained from freeze-thaw and agitation stress studies showed no significant changes in the attributes, indicating that the selected level of S-80 (0.25 mg / mL) in the final co-formulated composition is sufficient to protect against freeze-thaw and agitation induced particle formation and aggregation.

[0489] Example 10: Transparency and degree of opalescence of pharmaceutical products The results of the drug product transparency and opacity tests are shown in Tables 16-18. The color analysis results of the drug product under all conditions evaluated to date meet the acceptance criteria of "not more intense than reference Y5". At 5°C, no significant changes were observed in PS-80 content over the 9 months. At 25°C and 40°C, a trend toward a decrease was observed over the 6 months, but was within the specification. No changes in L-methionine were observed under any condition or at any time point throughout the study.

[0490] The 9-month data from HP-IEX showed no substantial change in acidic variants after storage at the proposed 5°C storage condition. At 25°C, there was a slight increase in % acidic variants, while at 40°C, a significant increase was observed through 6 months. For the anti-PD-1 monoclonal antibody, there was no change in % basic variants under all study conditions, while the anti-ILT4 / anti-PD-1 co-formulation showed a decrease through 6 months at 40°C. For the anti-ILT4 / anti-PD-1 co-formulation, there was a significant decrease in % major components at 40°C, while a smaller decrease was observed through 6 months for the anti-PD-1 monoclonal antibody. For the anti-ILT4 / anti-PD-1 co-formulation, a slight decrease was also observed at 25°C. No changes were observed for any of the components in the remaining conditions through 9 months. No changes were observed in % combined variants across conditions or time points for the formulations tested. Anti-PD-1 monoclonal antibody % major component decreased slightly through 6 months, with no changes observed in the remaining conditions with the study probe. No changes were observed in % mixed basic variants between conditions through 9 months for the formulations tested.

[0491] No changes were observed in % monomer and % high molecular weight species for formulations tested over 9 months at 5°C and 25°C. At 40°C, there was a gradual decrease over 6 months, with % monomer falling below the specification at 6 months. % high molecular weight species at 40°C exceeded the upper specification limit after 3 months.

[0492] For the formulations tested, an increase in % low molecular weight species was observed, falling below the specification limit after 2 months at 40°C, while no change was observed at 5°C and 25°C.

[0493] No significant changes in % intact IgG were observed for the formulations tested at 5° C. and 25° C. Results at 40° C. showed a decrease at 6 months and below specification after 3 months.

[0494] The formulations tested showed no change in % heavy + light chains at the recommended storage condition of 5° C. and at stressed conditions of 25° C. At 40° C., the % heavy + light chain results decreased and were below specification after 3 months. The formulations tested did not change over the study period across conditions and time points.

[0495] For the formulations tested, the number of visible particles was within specifications throughout the study period and under all conditions. Results from all analytical test panels were within acceptance criteria for up to 9 months under recommended storage conditions.

[0496] [Table 16] TIFF2025514174000022.tif245170TIFF2025514174000023.tif53169

[0497] [Table 17] TIFF2025514174000025.tif239169TIFF2025514174000026.tif50170

[0498] [Table 18] TIFF2025514174000028.tif235170

[0499] TIFF2025514174000029.tif62170

[0500] Example 11: Ion exchange (IEX) method for measuring acidic species of anti-PD-1 antibodies For the IEX method, a Waters Alliance LC system (Milford, MA, USA) was used, and Thermo Scientific's ProPac WCX-10 (p / n:054993, particle size 10um, diameter 4mm, length 250mm) was selected with a sample load of 80μg. The mobile phase (A) was 24mM MES (pH 6.1) (containing 4% acetonitrile), and the mobile phase (B) was 20mM sodium phosphate, 95mM NaCl (pH 8.0) (containing 4% acetonitrile) as a nonlinear sigmoidal pH gradient. Separation was performed at a flow rate of 0.5mL min−1. -1 The column was monitored at 35° C. for 34 min. The column temperature was 35° C. The gradient used was as follows: 0-0.6 min, 22%-22% B; 0.6-15.0 min, 22%-29% B; 15.0-30.0 min, 29%-70% B; 30.0-30.5 min, 70%-100% B; and 30.5-33.0 min, 100%-100% B. The mobile phase (C) was 10 mM CHES (pH 8.0), 40 mM Tris, 15 mM EDTA, 200 mM NaCl, and 4% acetonitrile, and 0.5 mL of mobile phase (D) was used. -1 Strip the column with 1.0 mL of ethanol at 34.5-44.5 min. -1 The flow rate was set to 0.5 mL min−1 for 44.5–45 min. -1 The eluate was monitored at 280 nm for peak detection. The assay variability was determined to be within 1%.

[0501] U.S. Provisional Application No. 63 / 143,461 is incorporated by reference in its entirety. All references cited herein are incorporated by reference to the same extent as if each individual publication, database entry (e.g., GenBank sequence or GeneID entry), patent application or patent was specifically and individually indicated to be incorporated by reference. Each and every individual publication, database entry (e.g., GenBank sequence or GeneID entry), patent application or patent is clearly identified in accordance with 37 CFR § 1.57(b)(2) even if such citation is not immediately adjacent to a dedicated statement that it is incorporated by reference. The inclusion of a presented statement of incorporation by reference herein in no way weakens this general statement of incorporation by reference, if such statement is present. The citation of a reference herein is not intended as an admission that the reference is pertinent prior art, nor does it imply any admission as to the contents or date of these publications or documents. In the event that a reference provides a definition for a claimed term that contradicts a definition set forth herein, the definition set forth herein shall be used in interpreting the claimed invention.

Claims

1. (i) about 10 mg / mL to about 200 mg / mL of an anti-human immunoglobulin-like transcript 4 (anti-ILT4) antibody or antigen-binding fragment thereof; (ii) about 10 mg / mL to about 200 mg / mL of an anti-PD-1 antibody or antigen-binding fragment thereof; (iii) about 5 mM to about 20 mM of a buffering agent; (iv) from about 6% to about 8% weight / volume (w / v) non-reducing sugars; (v) about 0.01% to about 0.10% (w / v) of a non-ionic surfactant; and (vi) about 1 mM to about 20 mM of an antioxidant; A formulation comprising: wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising: CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E, or D and X2 is S or A) (SEQ ID NO: 20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21); wherein the anti-PD-1 antibody, or antigen-binding fragment thereof, comprises a variable light chain region comprising a CDRL1 of SEQ ID NO: 95, a CDRL2 of SEQ ID NO: 96, and a CDRL3 of SEQ ID NO: 97; and a variable heavy chain region comprising a CDRH1 of SEQ ID NO: 100, a CDRH2 of SEQ ID NO: 101, and a CDRH3 of SEQ ID NO: 102; wherein the buffer is an L-histidine buffer, an acetate buffer, or a citrate buffer; wherein the non-reducing sugar is a disaccharide, wherein the nonionic surfactant is polysorbate 20 or polysorbate 80, and wherein the antioxidant is methionine or a pharma- ceutically acceptable salt thereof. The formulation.

2. 2. The formulation of claim 1, comprising: 40-60 mg / mL of the anti-ILT4 antibody; 15-35 mg / mL of the anti-PD-1 antibody; about 7% sucrose or trehalose; about 0.025% polysorbate 80 or polysorbate 20; about 3-30 mM L-histidine buffer at a pH of about 5.0-6.5; and about 5-20 mM methionine or a pharma-ceutically acceptable salt thereof.

3. 2. The formulation of claim 1, comprising: 45-55 mg / mL of the anti-ILT4 antibody; 20-30 mg / mL of the anti-PD-1 antibody; about 7% sucrose or trehalose; about 0.025% polysorbate 80 or polysorbate 20; about 3-30 mM L-histidine buffer at about pH 5.0-6.5; and about 10-20 mM L-methionine or a pharma-ceutically acceptable salt thereof.

4. 2. The formulation of claim 1, wherein the buffer is an L-histidine buffer.

5. The formulation according to any one of claims 1 to 4, wherein the non-reducing sugar is sucrose.

6. The formulation of any one of claims 1 to 5, wherein the non-ionic surfactant is polysorbate 80.

7. The formulation according to any one of claims 1 to 6, wherein the antioxidant is L-methionine.

8. The formulation comprises: (i) about 10 mg / mL to about 200 mg / mL of an anti-ILT4 antibody (anti-ILT4) or an antigen-binding fragment thereof; (ii) about 10 mg / mL to about 200 mg / mL of an anti-PD-1 antibody or antigen-binding fragment thereof; (iii) about 5 mM to about 20 mM L-histidine buffer; (iv) about 6% to about 8% weight / volume (w / v) sucrose; (v) about 0.01% to about 0.10% (w / v) polysorbate 80; and (vi) about 1 mM to about 20 mM L-methionine; A formulation comprising: wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising: CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E, or D and X2 is S or A) (SEQ ID NO: 20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21); and wherein the anti-PD-1 antibody, or antigen-binding fragment thereof, comprises a variable light chain region comprising a CDRL1 of SEQ ID NO:95, a CDRL2 of SEQ ID NO:96, and a CDRL3 of SEQ ID NO:97; and a variable heavy chain region comprising a CDRH1 of SEQ ID NO:100, a CDRH2 of SEQ ID NO:101, and a CDRH3 of SEQ ID NO:102; or a pharma- ceutically acceptable salt thereof. The formulation.

9. 9. The formulation of claim 8 comprising about 8 mM to about 12 mM L-histidine buffer.

10. 10. The formulation of claim 8 or 9, comprising about 5 mM to about 10 mM L-methionine.

11. 11. The formulation of any of claims 8 to 10, comprising about 0.01% to about 0.05% (w / v) polysorbate 80.

12. The formulation of any of claims 1 or 4-11, comprising about 10 mg / mL to about 150 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof.

13. 12. The formulation of any of claims 1 or 4-11, comprising about 10 mg / mL to about 150 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof.

14. 13. The formulation of claim 12, wherein the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 10 mg / mL, about 12.5 mg / mL, about 25 mg / mL, about 50 mg / mL, about 75 mg / mL, about 100 mg / mL, about 125 mg / mL or about 150 mg / mL.

15. 13. The formulation of claim 12, wherein the concentration of the anti-PD-1 antibody or antigen-binding fragment thereof is about 10 mg / mL, about 12.5 mg / mL, about 25 mg / mL, about 50 mg / mL, about 75 mg / mL, about 100 mg / mL, about 125 mg / mL, or about 150 mg / mL.

16. 14. The formulation of any of claims 1-13, comprising about 25 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

17. 14. The formulation of any of claims 1-13, comprising about 50 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 25 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

18. 14. The formulation of any of claims 1-13, comprising about 75 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 50 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

19. 14. The formulation of any of claims 1-13, comprising about 100 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 75 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

20. 14. The formulation of any of claims 1-13, comprising about 125 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 100 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

21. The formulation of any one of claims 1 to 20, wherein the formulation has a pH of about 5.0 to about 6.

8.

22. 22. The formulation of any one of claims 1 to 21, wherein the formulation has a pH of about 5.5 to about 6.

0.

23. 23. The formulation of any one of claims 1 to 22, wherein the formulation has a pH of about 5.

5.

24. A formulation of anti-human immunoglobulin-like transcript 4 ("anti-ILT4") antibody or antigen-binding fragment thereof, comprising: (i) about 50 mg / mL to about 100 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mg / mL to about 200 mg / mL of an anti-PD-1 antibody or antigen-binding fragment thereof; (iii) about 10 mM L-histidine buffer, pH about 5.5; (iv) about 7% weight / volume (w / v) sucrose; (v) about 0.025% (w / v) polysorbate 80; and (vi) about 10 mM L-methionine; 1. A formulation of an anti-human immunoglobulin-like transcript 4 (anti-ILT4) antibody or antigen-binding fragment thereof comprising: wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising: CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E, or D and X2 is S or A) (SEQ ID NO: 20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21); and wherein the anti-PD-1 antibody, or antigen-binding fragment thereof, comprises a variable light chain region comprising a CDRL1 of SEQ ID NO: 95, a CDRL2 of SEQ ID NO: 96, and a CDRL3 of SEQ ID NO: 97; and a variable heavy chain region comprising a CDRH1 of SEQ ID NO: 100, a CDRH2 of SEQ ID NO: 101, and a CDRH3 of SEQ ID NO: 102; The formulation.

25. 25. The formulation of any one of claims 1 to 24, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising: CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E or D and X2 is S or A) (SEQ ID NO: 20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

26. 25. The formulation of any one of claims 1 to 24, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising: CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

27. The formulation of any one of claims 1 to 24, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO: 57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:

58.

28. The formulation of any one of claims 1 to 24, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2; and a light chain comprising the amino acid sequence set forth in SEQ ID NO:

7.

29. The formulation of any one of claims 1 to 24, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:80; and a light chain comprising the amino acid sequence set forth in SEQ ID NO:

7.

30. The formulation of any one of claims 1 to 24, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO: 57; and a light chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:

58.

31. The formulation of any one of claims 1 to 24, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 2; and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:

7.

32. The formulation of any one of claims 1 to 24, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 80; and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:

7.

33. The formulation of any one of claims 1 to 24, wherein the anti-human PD-1 antibody or antigen-binding fragment thereof comprises a variable light chain region comprising the amino acid sequence set forth in SEQ ID NO: 98; and a variable heavy chain region comprising the amino acid sequence set forth in SEQ ID NO:

103.

34. The formulation of any one of claims 1 to 24, wherein the anti-human PD-1 antibody or antigen-binding fragment thereof comprises a variable light chain region comprising the amino acid sequence set forth in SEQ ID NO: 99; and a variable heavy chain region comprising the amino acid sequence set forth in SEQ ID NO:

104.

35. The formulation of any one of claims 1 to 24, wherein the anti-human PD-1 antibody or antigen-binding fragment thereof is composed of two light chains and two heavy chains, wherein the two light chains consist of the amino acid sequence set forth in SEQ ID NO: 99, and the two heavy chains consist of the amino acid sequence set forth in any one of SEQ ID NOs: 104-109.

36. The formulation of any one of claims 1 to 35, wherein the anti-human PD-1 antibody is pembrolizumab.

37. The formulation of any one of claims 1 to 36, wherein the ratio of anti-ILT4 antibody to anti-PD-1 antibody is 1:1 or 1:

2.

38. After 9 months at 5°C, (i) the percent monomer of the anti-ILT4 antibody is ≧95% as measured by size exclusion chromatography; (ii) the % heavy chain and % light chain of said anti-ILT4 antibody are ≧90% as measured by reducing CE-SDS; (iii) the % heavy chain and % light chain of the anti-ILT4 antibody are ≧95% as measured by reducing CE-SDS; (iv) the percent intact IgG of the anti-ILT4 antibody is > 90% when measured by non-reducing CE-SDS; and / or (v) the percent intact IgG of the anti-ILT4 antibody is >= 95% when measured by non-reducing CE-SDS; A formulation according to any one of claims 1 to 37.

39. 39. The formulation of any one of claims 1 to 38, wherein the formulation is a liquid formulation stored at about 3°C ​​to about 5°C or a reconstituted solution from a lyophilized formulation.

40. After storage at about 3°C ​​to about 5°C for up to 9 months: (i) the percent monomer of the anti-ILT4 antibody, or antigen-binding fragment thereof, is at least about 99% as measured by ultra-performance size exclusion chromatography; (ii) O.D. ３５０－５００ The turbidity of the formulation is up to about 0.135 when measured at (iii) the % main peak of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 63%, the % acidic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 23%, and / or the % basic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 14%, as measured by high performance ion exchange chromatography; (iv) the number of sub-visible particles having a size of at least 2 μm is at most about 3500, as measured by microflow imaging; and / or (v) the percent oxidation of one or more amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% as measured by reduced peptide mapping; A formulation according to any one of claims 1 to 24.

41. The formulation of any one of claims 1-34 or 37-41, wherein the anti-PD-1 antibody or antigen-binding fragment thereof is a monoclonal antibody.

42. 42. A method of treating cancer in a human patient in need thereof comprising administering an effective amount of the formulation of any one of claims 1-41.

43. 43. The method of claim 42, wherein the cancer is selected from the group consisting of colorectal cancer, esophageal cancer, melanoma, non-small cell lung cancer, ovarian cancer, renal cell carcinoma and small cell lung cancer.

44. Use of a formulation according to any one of claims 1 to 41 for the preparation of a medicament for treating cancer.

45. Use of a formulation according to any one of claims 1 to 42 for treating cancer in a human patient.

46. 46. ​​The use according to claim 45, wherein the cancer is selected from the group consisting of colorectal cancer, esophageal cancer, melanoma, non-small cell lung cancer, ovarian cancer, renal cell carcinoma and small cell lung cancer.

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