Formulations of Anti-CD38 antibodies for subcutaneous administration

Abstract

To provide formulations of anti-CD38 antibodies suitable for subcutaneous administration to a subject in need thereof.SOLUTION: Formulations include a high concentration of antibody, a viscosity lowering agent, a stabilizing agent, a buffering agent, and a surfactant. In certain embodiments, the viscosity of the solution is at most 25 mPa s, and the pH of the solution is 5.9-7.0. In certain embodiments, the anti-CD38 antibody is isatuximab. The formulations are useful in treating CD38+ hematological malignancies, including multiple myeloma, as well as autoimmune and inflammatory diseases, in humans.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Related Applications This application claims priority to U.S. Provisional Patent Application No. 62 / 944,082, filed December 5, 2019, the entire disclosure of which is hereby incorporated by reference.

[0002] Sequence Listing This application contains a Sequence Listing that has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy, created on December 4, 2020, is named 712533_SA9-295PC_ST25.txt and is 9,675 bytes in size.

[0003] The present disclosure relates to formulations of antibodies useful in the treatment of diseases. More specifically, it relates to cancers, including multiple myeloma, as well as CD38 + It relates to formulations of anti-CD38 antibodies suitable for use in subcutaneous administration to treat other diseases and conditions in which cells play a role. [Background technology]

[0004] CD38 is a 45 kDa type II transmembrane glycoprotein with a long C-terminal extracellular domain and a short N-terminal cytoplasmic domain. + It is a bifunctional ectoenzyme that can catalyze the conversion of α-ADP to cyclic ADP-ribose (cADPR) and can also hydrolyze cADPR to ADP-ribose.

[0005] CD38 is upregulated in many hematological malignancies and cell lines derived from various hematological malignancies. Furthermore, most early pluripotent stem cells in the hematological system express CD38 - CD38 expression in hematological malignancies and its correlation with disease progression in chronic lymphocytic leukemia (CLL) makes CD38 an attractive target for antibody therapy.

[0006] CD38 +The cells have also been reported to be associated with a variety of other diseases and conditions, including many autoimmune diseases, such as rheumatoid arthritis and lupus erythematosus, as well as lipopolysaccharide (LPS)- or sepsis-induced acute kidney injury (Non-Patent Document 1).

[0007] Anti-CD38 antibodies that specifically recognize CD38 have been previously described, for example, in Patent Document 1. However, these antibodies have not been used as single agents to inhibit CD38. + When incubated with expressing cells, it is unable to induce apoptosis.

[0008] Monoclonal anti-CD38 antibodies are described in US Pat. No. 5,623,999.

[0009] The use of these specific anti-CD38 antibodies in combination with cytotoxic agents such as cytarabine, vincristine, cyclophosphamide and melphalan has been reported in US Pat. No. 5,623,299, ... and US Pat. No. 5,623,299.

[0010] Patent documents 7, 8, 9, 10, and 11 also describe the use of a humanized version of 38SB19 (also known as SAR650984 or isatuximab). A phase 3 clinical trial (NCT02990338) of isatuximab in combination with pomalidomide and dexamethasone for the treatment of patients with relapsed / refractory multiple myeloma recently met its primary endpoint, prolonging progression-free survival. [Prior art documents] [Patent documents]

[0011] [Patent Document 1] International Patent Application WO2006 / 099875 [Patent Document 2] International Patent Application WO2008 / 047242 [Patent Document 3] International Patent Application WO2010 / 061357 [Patent Document 4] International Patent Application WO2010 / 061358 [Patent Document 5] International Patent Application WO2010 / 061359 [Patent Document 6] International Patent Application WO2010 / 061360 [Patent Document 7] International Patent Application WO2015 / 066450 [Patent Document 8] International Patent Application WO2012 / 076663 [Patent Document 9] International Patent Application WO2014 / 089416 [Patent Document 10] International Patent Application WO2014 / 159911 [Non-patent literature]

[0012] [Non-Patent Document 1] Shu B et al. Cell Signal (2018) 42:249-58 Summary of the Invention [Means for solving the problem]

[0013] Embodiments of the present disclosure relate to formulations of anti-CD38 antibodies suitable for subcutaneous administration to a subject. Advantageously, the formulations disclosed herein are suitable for subcutaneous administration by either injection or infusion, including large-volume subcutaneous infusion. The formulations can be used in the treatment of diseases or conditions characterized by CD38-expressing cells. Such diseases and conditions include, but are not limited to, CD38-expressing solid tumors, such as prostate cancer, various hematological malignancies, such as non-Hodgkin's lymphoma (NHL), multiple myeloma (MM), acute myeloid leukemia (AML), acute lymphoblastic leukemia (B-cell ALL), and / or chronic lymphocytic leukemia (CLL). Such diseases and conditions further include, but are not limited to, autoimmune diseases, including rheumatoid arthritis and systemic lupus erythematosus, and lipopolysaccharide (LPS)- or sepsis-induced acute kidney injury.

[0014] In certain embodiments, the present disclosure relates to formulations of anti-CD38 antibodies comprising a high concentration of antibody, a pH of 5.9 to 7.0, and a viscosity of at most 25 mPa·s at 20°C.

[0015] In certain embodiments, the present disclosure relates to a formulation comprising at least 100 mg / mL of an anti-CD38 antibody, a viscosity-reducing agent, a stabilizer, a buffer, and a surfactant, and having a pH of 5.9 to 7.0 and a viscosity of at most 25 mPa·s at 20°C.

[0016] One embodiment of the present disclosure provides a formulation comprising at least 100 mg / mL of an anti-CD38 antibody, The anti-CD38 antibody comprises a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR2-H2, and CDR-H3, each having the amino acid sequences set forth in SEQ ID NOS: 1 to 3, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, each having the amino acid sequences set forth in SEQ ID NOS: 4 to 6, The formulation comprises a viscosity-reducing agent, a stabilizer, a buffer and a surfactant; The formulation has a pH of 5.9 to 7.0 and a viscosity of at most 25 mPa·s at 20°C.

[0017] In one particular embodiment, the viscosity-reducing agent is 90-150 mM Arg-Cl.

[0018] In one particular embodiment, the viscosity-reducing agent is 90-125 mM Arg-Cl.

[0019] In one particular embodiment, the viscosity-reducing agent is 110 mM Arg-Cl.

[0020] In one particular embodiment, the surfactant is poloxamer 188.

[0021] In one particular embodiment, the surfactant is 0.4% (w / v) poloxamer 188.

[0022] In certain embodiments, the buffering agent is histidine.

[0023] In one particular embodiment, the buffering agent is 9 mM histidine.

[0024] In certain embodiments, the viscosity-reducing agent is 90-150 mM Lys-Ac.

[0025] In one particular embodiment, the viscosity-reducing agent is 125 mM Lys-Ac.

[0026] In certain embodiments, the surfactant is polysorbate 80.

[0027] In one particular embodiment, the surfactant is 0.04% (w / v) polysorbate 80.

[0028] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody.

[0029] In certain embodiments, the formulation comprises 140 mg / mL of anti-CD38 antibody.

[0030] In certain embodiments, the stabilizer is sucrose.

[0031] In certain embodiments, the stabilizer is 2% (w / v) sucrose.

[0032] In certain embodiments, the pH is between 5.9 and 7.0.

[0033] In certain embodiments, the pH is between 5.9 and 6.5.

[0034] One embodiment of the present disclosure is a formulation comprising 140 mg / mL of an anti-CD38 antibody, wherein the anti-CD38 antibody comprises a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences set forth in SEQ ID NOS: 1 to 3, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences set forth in SEQ ID NOS: 4 to 6, The formulation contained 9 mM histidine, 110 mM Arg-Cl, 2 (w / v)% sucrose and 0.4 (w / v)% poloxamer 188; The formulation has a pH of 6.2 and a viscosity of at most 14 mPa·s at 20°C.

[0035] One embodiment of the present disclosure is a formulation comprising 140 mg / mL of an anti-CD38 antibody, wherein the anti-CD38 antibody comprises a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences set forth in SEQ ID NOS: 1 to 3, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences set forth in SEQ ID NOS: 4 to 6, The formulation contained 125 mM Lys-Ac, 2 (w / v)% sucrose and 0.04 (w / v)% polysorbate 80, The formulation has a pH of 6.2 and a viscosity of at most 14 mPa·s at 20°C. do.

[0036] In certain embodiments, the formulation is suitable for subcutaneous administration, either by injection or infusion, including bolus subcutaneous infusion.

[0037] In certain embodiments, the VH of the anti-CD38 antibody comprises the amino acid sequence set forth in SEQ ID NO:7, and the VL of the anti-CD38 antibody comprises the amino acid sequence set forth in SEQ ID NO:8.

[0038] In certain embodiments, the anti-CD38 antibody is isatuximab.

[0039] Certain aspects of the present disclosure relate to packaged pharmaceutical products comprising a sterile container containing a therapeutically effective amount of a formulation of the present disclosure.

[0040] Certain aspects of the present disclosure relate to devices comprising a therapeutically effective amount of the formulation of the present disclosure.

[0041] In certain embodiments, the device may be, for example, a syringe containing the formulation, a syringe driver, and an infusion pump.

[0042] In certain embodiments, the syringe is a pre-filled syringe.

[0043] Certain aspects of the present disclosure include CD38 + A method of treating a disease or condition characterized by the presence or activity of a cell, comprising administering to a subject in need thereof an effective amount of a formulation of the present disclosure, wherein the formulation is administered subcutaneously.

[0044] In certain embodiments, CD38 + Diseases or conditions characterized by the presence or activity of CD38 cells + It is a hematological malignancy.

[0045] In certain embodiments, CD38 + The disease or condition characterized by the presence or activity of the cells is an autoimmune or inflammatory disease or condition.

[0046] Certain aspects of the present disclosure include CD38 + A method of treating a hematological malignancy, comprising administering to a subject in need thereof an effective amount of a formulation of the present disclosure, wherein the formulation is administered subcutaneously.

[0047] Certain aspects of the present disclosure include CD38 + 1. A method of treating a hematological malignancy, comprising administering to a subject in need thereof an effective amount of a formulation of an anti-CD38 antibody comprising at least 100 mg / mL of anti-CD38 antibody; The anti-CD38 antibody comprises a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, each having the amino acid sequences set forth in SEQ ID NOS: 1 to 3, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, each having the amino acid sequences set forth in SEQ ID NOS: 4 to 6, The formulation comprises a viscosity-reducing agent, a stabilizer, a buffer and a surfactant; The formulation has a pH of 5.5 to 7.0 and a viscosity of at most 25 mPa·s at 20°C, and the formulation is administered subcutaneously.

[0048] Certain aspects of the present disclosure include CD38 + 1. A method of treating a hematological malignancy, comprising administering to a subject in need thereof an effective amount of an anti-CD38 formulation comprising 140 mg / mL of an anti-CD38 antibody; The anti-CD38 antibody has a variable heavy chain region including three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, each of which contains the amino acid sequences shown in SEQ ID NOS: 1 to 3. a variable light chain region (VH) comprising three CDRs, CDR-L1, CDR-L2 and CDR-L3, each having the amino acid sequences shown in SEQ ID NOs: 4 to 6; The formulation contained 9 mM histidine, 110 mM Arg-Cl, 2 (w / v)% sucrose and 0.4 (w / v)% poloxamer 188; The formulation has a pH of 6.2 and a viscosity of at most 14 mPa·s at 20° C., and the formulation is administered subcutaneously.

[0049] In certain embodiments, the method comprises administering to a subject in need thereof an effective amount of a formulation comprising 140 mg / mL of an anti-CD38 antibody; The anti-CD38 antibody comprises a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, each having the amino acid sequences set forth in SEQ ID NOS: 1 to 3, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, each having the amino acid sequences set forth in SEQ ID NOS: 4 to 6, The formulation contained 9 mM histidine, 110 mM Arg-Cl, 2 (w / v)% sucrose and 0.4 (w / v)% poloxamer 188; The formulation has a pH of 6.3 and a viscosity of at most 14 mPa·s at 20° C., and the formulation is administered subcutaneously.

[0050] In certain embodiments, the formulation is administered by subcutaneous injection.

[0051] In certain embodiments, the formulation is administered by subcutaneous injection.

[0052] In certain embodiments, the subcutaneous injection is a large volume subcutaneous injection, for example, >2 mL to 30 mL.

[0053] In certain embodiments, CD38 + The method for treating hematological malignancies comprises administering to a subject a therapeutically effective amount of C38 + The method further includes administering to the subject one or more additional agents suitable for treating the hematological cancer. In some embodiments, the other agents are, for example, corticosteroids (e.g., dexamethasone), chemotherapeutic agents, proteasome inhibitors, immunomodulatory agents, or combinations thereof.

[0054] In certain embodiments, the chemotherapeutic agent is, for example, cytarabine, daunorubicin, daunomycin, doxorubicin, liposomal doxorubicin, idarubicin, mitoxantrone, gemtuzumab, clofarabine, cladribine, hydroxyurea, etoposide, amsacrine, an FLT3 inhibitor such as gilteritinib, 5-azacytidine, decitabine, melphalan, cyclophosphamide, or vincristine, or a combination thereof.

[0055] In certain embodiments, the immunomodulatory agent is, for example, thalidomide, lenalidomide, or pomalidomide, or a combination thereof.

[0056] In certain embodiments, the proteasome inhibitor is, for example, ixazomib, carfilzomib, or bortezomib, or a combination thereof.

[0057] In certain embodiments, CD38 + A method of treating a hematological malignancy includes administering to a subject isatuximab formulated for subcutaneous administration as defined herein and two or more additional agents from different classes of compounds, such as immunomodulators or proteasome inhibitors.

[0058] In certain embodiments, CD38 + The hematological malignancy is multiple myeloma. In embodiments, the multiple myeloma is relapsed / refractory multiple myeloma. In some embodiments, the patient has received at least two prior therapies for multiple myeloma, including lenalidomide and a proteasome inhibitor, and has shown disease progression during or after completion of the last therapy. [Brief explanation of the drawings]

[0059] [Figure 1] 1 is a series of graphs showing the number of subvisible particles ≧10 μm and ≧25 μm after heat stress (top panel) and shaking stress (bottom panel) of isatuximab in the indicated buffer systems: 2w 40°C, 2 weeks at 40°C; 4w 40°C, 4 weeks at 40°C; Cit, citrate buffer; His, histidine buffer; Pho, phosphate buffer; Ace, acetate buffer. [Figure 2] Graph showing the percentage of soluble aggregates (high molecular weight aggregates (HMW) as measured using size exclusion chromatography (SE-HPLC)) after heat stress at 40°C. 2w 40°C, 2 weeks at 40°C; 4w 40°C, 4 weeks at 40°C; Cit, citrate buffer; His, histidine buffer; Pho, phosphate buffer; Ace, acetate buffer. [Figure 3]1 is a graph depicting the percentage of soluble aggregates (high molecular weight aggregates (HMW)) as measured using size exclusion chromatography (SE-HPLC) after heat stress at 40° C. in histidine buffers with the indicated pH and concentration values: 1M 40° C., 1 month at 40° C. [Figure 4] 1 is a graph depicting the percentage acidic form of isatuximab as measured using a weak cation exchange assay after shaking stress at 40° C. in histidine buffers with the indicated pH and concentration values: 1M 40° C., 1 month at 40° C. [Figure 5] 1 is a graph depicting the viscosity of 200 mg / mL isatuximab in arginine-Cl pH 6.0 at the concentrations indicated. [Figure 6A] 1 is a graph depicting the viscosity of 150 mg / mL isatuximab over a range of concentrations in arginine-HCl buffer pH 6.3. The inset shows details at higher concentrations of arginine-HCl. [Figure 6B] 1 is a graph showing the viscosity of 180 mg / mL isatuximab over a range of concentrations in arginine-HCl buffer pH 6.3. The inset shows details at higher concentrations of arginine-HCl. [Figure 7A] 1 is a graph depicting the viscosity of 150 mg / mL isatuximab over a range of pH in 150 mM arginine-HCl buffer. [Figure 7B] 1 is a graph depicting the viscosity of 150 mg / mL isatuximab over a range of buffer pH in 200 mM arginine-HCl. [Figure 8] Graph showing viscosity plotted as a function of mAb concentration at pH 5.5, 5.9, 6.2 and 7.0 (T=20° C.). Goodness of fit: Mooney-based equation. [Figure 9] 1 is a graph depicting viscosity plotted as a function of pH at mAb concentrations of 126, 140, 147, and 154 g / L (T=20° C.). [Figure 10]1 is a graph depicting viscosity plotted as a function of temperature at mAb concentrations of 126, 143, and 154 g / L (pH=6.2). [Figure 11] 1 is a graph depicting viscosity plotted as a function of temperature at mAb concentrations of 142 and 152 g / L (pH=5.9). [Figure 12] 1 is a graph depicting viscosity plotted as a function of arginine concentration at a mAb concentration of 140 g / L (pH=6.2, T=20° C.). [Figure 13] Graph showing relative monomer content detected using HP-SEC analysis of isatuximab in formulations F4-1 to F4-16 (Run 1 to Run 16, respectively, n=2, average); T0: no treatment, T-mech: mechanical stress, T-5×FT: 5 freeze / thaw cycles, T-1m_40℃: 1 month at 40℃. [Figure 14] Graph showing relative monomer content detected using HP-SEC analysis of isatuximab in formulations F10-1 to F10-16 (Run 1 to Run 16, respectively, n=2, average); T0: no treatment, T-mech: mechanical stress, T-5×FT: 5 freeze / thaw cycles, T-1m_40°C: 1 month at 40°C. [Figure 15] Graph showing the relative content of total aggregates (HMWS) detected using HP-SEC analysis of isatuximab in formulations F4-1 to F4-16 (Run 1 to Run 16, respectively, n=2, average); T0: no treatment, T-mech: mechanical stress, T-5×FT: 5 freeze / thaw cycles, T-1m_40°C: 1 month at 40°C. [Figure 16] Figure 1 shows the relative content of total aggregates (HMWS) detected using HP-SEC analysis of isatuximab in formulations F10-1 to F10-16 (Run 1 to Run 16, respectively, n=2, average); T0: no treatment, T-mech: mechanical stress, T-5xFT: 5 freeze / thaw cycles, T-1m_40°C: 1 month at 40°C. [Figure 17]Graph showing the relative content of all fragments (LMWS) detected using HP-SEC analysis of isatuximab in formulations F4-1 to F4-16 (Run 1 to Run 16, respectively, n=2, average); T0: no treatment, T-mech: mechanical stress, T-5×FT: five freeze / thaw cycles, T-1m_40℃: 1 month at 40℃. [Figure 18] Graph showing the relative content of all fragments (LMWS) detected using HP-SEC analysis of isatuximab in formulations F10-1 to F10-16 (Run 1 to Run 16, respectively, n=2, average); T0: no treatment, T-mech: mechanical stress, T-5×FT: 5 freeze / thaw cycles, T-1m_40℃: 1 month at 40℃. [Figure 19] 1 is a graph showing the acidic peak content obtained from capillary isoelectric focusing (cIEF) analysis of isatuximab in formulations F4-1 to F4-16 (Run 1 to Run 16, respectively, n=2, average); T0: no treatment, T-1m_40℃: 1 month at 40℃. [Figure 20] 1 is a graph showing the acidic peak content obtained from cIEF analysis of isatuximab in formulations F10-1 to F10-16 (Run 1 to Run 16, respectively, n=2, average); T0: no treatment, T-1m_40℃: 1 month at 40℃. [Figure 21] 1 is a graph depicting the relative area loss [%] of the monomer peak content obtained from cIEF analysis of isatuximab in formulations F4-1 to F4-16 and F10-1 to F10-16 (runs 1 to 16, respectively, n=2, average) after 1 month of storage at 40° C. / 75% rh. [Figure 22] 1 is a graph depicting plasma cortisol in minipigs subcutaneously injected with the indicated formulations of isatuximab or NaCl control as described in Example 5. [Figure 23] 1 is a graph depicting plasma substance P in minipigs subcutaneously injected with the indicated formulations of isatuximab or NaCl control as described in Example 5. [Figure 24]1 is a graph depicting serum concentrations of isatuximab over time in minipigs 1-5 of Group I as described in Example 6. [Figure 25] 1 is a graph depicting serum concentrations of isatuximab over time in minipigs 6-10 of Group II as described in Example 6. [Figure 26] 1 is a graph depicting serum concentrations of isatuximab over time in minipigs 11-15 of Group III as described in Example 6. [Figure 27] 1 is a graph depicting serum concentrations of isatuximab over time in minipigs 16-20 of Group IV as described in Example 6. DETAILED DESCRIPTION OF THE INVENTION

[0060] Provided herein are formulations of anti-CD38 antibodies that are suitable for subcutaneous administration to a subject. Advantageously, the formulations disclosed herein are suitable for subcutaneous administration by either injection or infusion, including large-volume subcutaneous infusion. The formulations can be used in the treatment of diseases or conditions characterized by CD38-expressing cells. Such diseases and conditions include, but are not limited to, various hematological malignancies, such as non-Hodgkin's lymphoma (NHL), multiple myeloma (MM), acute myeloid leukemia (AML), acute lymphoblastic leukemia (B-cell ALL), and / or chronic lymphocytic leukemia (CLL). ... Further examples include, but are not limited to, autoimmune diseases, including rheumatoid arthritis and systemic lupus erythematosus, and lipopolysaccharide (LPS)- or sepsis-induced acute kidney injury. In certain embodiments, the formulations provided herein comprise a high concentration of antibody, a pH of 5.5-7.0, and a viscosity of at most 25 mPa·s at 20°C. In some embodiments provided herein, the formulations are aqueous.

[0061] A "hematologic malignancy" is a type of cancer that affects the blood, bone marrow, and lymph nodes. Because the three are closely linked through the immune system, a disease affecting one of the three may affect the others as well. Hematologic malignancies include non-Hodgkin's lymphoma (NHL) (including, for example, Burkitt's lymphoma (BL) and T-cell lymphoma (TCL)), multiple myeloma (MM), chronic lymphocytic leukemia (CLL) (such as, for example, B-cell chronic lymphocytic leukemia (B-CLL) and hairy cell leukemia (HCL)), B-cell and T-cell acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), Hodgkin's lymphoma (HL), and chronic myeloid leukemia (CML). In some embodiments, the hematologic malignancy is CD38 + It is a hematological malignancy.

[0062] Therefore, "CD38 + Hematologic malignancies are those in which cancer cells express CD38, as described above. + The cells have also been reported to be involved in many autoimmune and inflammatory diseases and disorders, including rheumatoid arthritis and systemic lupus erythematosus, as well as other conditions, including LPS- or sepsis-induced acute kidney injury.

[0063] CD38 + Hematological malignancies include B-cell non-Hodgkin's lymphoma (NHL), multiple myeloma (MM), acute myeloid leukemia (AML), acute lymphoblastic leukemia (B-cell ALL), and / or chronic lymphocytic leukemia (CLL). + The hematological malignancy is MM. + The hematological malignancy is relapsed and / or refractory multiple myeloma.

[0064] An "antibody" may be a natural or conventional antibody in which two heavy chains are linked to each other by disulfide bonds, and each heavy chain is linked to a light chain by a disulfide bond. There are two types of light chains, lambda (λ) and kappa (κ). There are five major heavy chain classes (or isotypes) that determine the functional activity of antibody molecules: IgM, IgG, IgA, IgD, and IgE. Furthermore, immunoglobulin subclasses (or subisotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2, are well characterized and are known to confer functional specialization. Each chain contains distinct sequence domains. The light chain contains two domains or regions, a variable domain (VL) and a constant domain (CL). The heavy chain contains four domains, a variable domain (VH) and three constant domains (CH1, CH2, and CH3, collectively referred to as CH). The variable regions of both the light (VL) and heavy (VH) chains determine antigen binding recognition and specificity. The constant region domains of the light (CL) and heavy (CH) chains confer important biological properties, such as antibody chain association, secretion, transplacental mobility, complement fixation, and Fc receptor (FcR) binding. The Fv fragment is the N-terminal portion of an immunoglobulin Fab fragment, containing the variable portions of one light chain and one heavy chain. Antibody specificity resides in the structural complementarity between the antibody binding site and an antigenic determinant. The antibody binding site is primarily composed of residues from the hypervariable or complementarity-determining regions (CDRs). Occasionally, residues from non-hypervariable or framework regions (FRs) influence the overall domain structure and therefore the binding site.

[0065] "Complementarity-determining region" or "CDR" refers to the amino acid sequences that together define the binding specificity and affinity of the natural Fv region of a natural immunoglobulin binding site. The light and heavy chains of an immunoglobulin each have three CDRs, designated CDR-L1, CDR-L2, CDR-L3, and CDR-H1, CDR-H2, CDR-H3, respectively. Conventional antibodies An antigen-binding site therefore comprises six CDRs, comprising a set of CDRs from each of the heavy and light chain V regions.

[0066] The definition of the CDR / FR for an immunoglobulin light or heavy chain is given based on the Kabat definition (worlwideweb.bioinf.org.uk / abs / ).

[0067] The antibody may be a non-naturally occurring antibody, such as a monoclonal antibody, a chimeric antibody, or a humanized antibody. As used herein, the term "monoclonal antibody" or "mAb" refers to an antibody molecule of a single amino acid composition directed against a specific antigen and should not be construed as requiring the production of the antibody by any particular method. Monoclonal antibodies can be produced by a single clone of a B cell or hybridoma, but can also be recombinant, i.e., produced by protein engineering.

[0068] The term "humanized antibody" refers to an antibody that is originally wholly or partially of non-human origin and that has been modified, for example, by replacing certain amino acids in the framework regions of the heavy and light chains, to avoid or minimize an immune response in humans. The constant domains of a humanized antibody can be, for example, human CH and CL domains. In certain embodiments, a humanized antibody has constant domains of human origin.

[0069] In some embodiments, an anti-CD38 antibody according to the present disclosure comprises a heavy chain comprising a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:1, a CDR-H2 comprising the amino acid sequence SEQ ID NO:2, and a CDR-H3 comprising the amino acid sequence SEQ ID NO:3, and a light chain comprising a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:4, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:5, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:6. CDR-H1 DYWMQ (SEQ ID NO: 1) CDR-H2 TIYPGDGDTGYAQKFQG (SEQ ID NO: 2) CDR-H3 GDYYGSNSLDY (SEQ ID NO: 3) CDR-L1 KASQDVSTVVA (SEQ ID NO: 4) CDR-L2 SASYRYI (SEQ ID NO: 5) CDR-L3 QQHYSPPYT (SEQ ID NO: 6)

[0070] In some embodiments, the antibody comprises a heavy chain variable domain (VH) comprising the amino acid sequence set forth in SEQ ID NO:7.

[0071] In some embodiments, the antibody comprises a light chain variable domain (VL) comprising the amino acid sequence set forth in SEQ ID NO:8.

[0072] In some embodiments, the antibody comprises a heavy chain variable domain (VH) comprising the amino acid sequence set forth in SEQ ID NO:7, and a light chain variable domain (VL) comprising the amino acid sequence set forth in SEQ ID NO:8. QVQLVQSGAEVAKPGTSVKLSCKASGYTFTDYWMQWVKQRPGQGLEWIGTIYPGDGDTGYAQKFQGKATLTADKSSKTVYMHLSSLASEDSAVYYCARGDYYGSNSLDYWGQGTSVTVSS (SEQ ID NO: 7) DIVMTQSHLSMSSTSLGDPVSITCKASQDVSTVVAWYQQKPGQSPRRLIYSASYRYIGVPDRFTGSGAGTDFTFTISSVQAEDLAVYYCQQHYSPPYTFGGGTKLEIKR (SEQ ID NO: 8)

[0073] In some embodiments, the anti-CD38 antibody according to the present disclosure is isatuximab. The heavy chain (HC) of isatuximab comprises the amino acid sequence set forth in SEQ ID NO:9, and the light chain (LC) of isatuximab comprises the amino acid sequence set forth in SEQ ID NO:10. QVQLVQSGAEVAKPGTSVKLSCKASGYTFTDYWMQWVKQRPGQGLEWIGTIYPGDGDTGYAQKFQGKATLTADKSSKTVYMHLSSLASEDSAVYYCARGDYYGSNSLDYWGQ GTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 9) DIVMTQSHLSMSSTSLGDPVSITCKASQDVSTVVAWYQQKPGQSPRRLIYSASYRYIGVPDRFTGSGAGTDFTFTISSVQAEDLAVYYCQQHYSPPYTFGGGTKLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 10)

[0074] In some embodiments, the antibody is CD38 + For use in the treatment of hematological malignancies, such as multiple myeloma (MM), including relapsed and / or refractory MM, or in MM patients who have received one or more prior therapies for MM.

[0075] In the context of the present disclosure, the term "treat" or "treatment" as used herein means to ameliorate, alleviate, inhibit the progression of, or prevent the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition. In certain embodiments, the term "treat" or "treatment" as used herein means to ameliorate, alleviate, or inhibit the progression of the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition.

[0076] As used herein, the term "CD38 + Treating hematologic malignancies: tumor CD38 + Malignant cell proliferation and / or CD38 + It refers to the inhibition of the progression of metastases from a tumor. Such treatment can also lead to regression of tumor growth, i.e., a decrease in the size of a measurable tumor.

[0077] In the context of this disclosure, a "therapeutically effective amount" of an antibody is an amount of the CD38 + It means a sufficient amount of antibody to treat a hematological malignancy.

[0078] In certain embodiments, the therapeutically effective amount of antibody administered subcutaneously to a subject is a dose of 500 mg to 2000 mg of antibody per dose.

[0079] In certain embodiments, the therapeutically effective amount of antibody administered to a subject is 1000 mg of antibody per dose. In certain embodiments, the therapeutically effective amount of antibody administered to a subject is 1400 mg of antibody per dose. In certain embodiments, the therapeutically effective amount of antibody administered to a subject is 1600 mg of antibody per dose.

[0080] As used herein, the term "subject" refers to a mammal. In certain embodiments, the term "subject" refers to a human.

[0081] The antibodies of the disclosure can be administered once weekly (QW), once every two weeks (Q2W), or a combination of once weekly and once every two weeks. In some embodiments, the antibodies are administered once every four weeks.

[0082] For example, the antibody can be administered to a subject at a dose of 500 mg to 1400 mg once weekly for four weeks (Cycle 1), and then once every other week (e.g., on days 1 and 15 of each subsequent four-week cycle).

[0083] In some embodiments, 1000 mg of antibody is administered to a subject once a week for 4 weeks (Cycle 1), and then 1000 mg of antibody is administered to a subject on days 1 and 15 of each subsequent 4-week cycle.

[0084] In some embodiments, 1000 mg of antibody is administered to a subject once every two weeks.

[0085] In some embodiments, 1400 mg of antibody is administered to a subject once a week for 4 weeks (Cycle 1), and then 1400 mg of antibody is administered to a subject on days 1 and 15 of each subsequent 4-week cycle.

[0086] In some embodiments, 1400 mg of antibody is administered to a subject once every two weeks. In some embodiments, the antibody can be administered according to an intermittent regimen with an interval of one or two weeks between each dose, which can be one to two weeks longer depending on tolerance to the previous dose.

[0087] As used herein, a "cycle" refers to four calendar weeks, or 28 days. "Weekly" administration means once every seven days. "Biweekly" administration means once every 14 days. "Once per cycle" or "every four weeks" administration means once every 28 days.

[0088] In some embodiments, the number of cycles of antibody administration may be between 2 and 50, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 25, 30, 35, 45, or 50 cycles.

[0089] Formulations of the Invention The present disclosure provides certain formulations of anti-CD38 antibodies. In some embodiments, the formulations are liquid formulations. In certain embodiments, such formulations (antibody formulations) are suitable for administration to a subject in need of treatment with an anti-CD38 antibody.

[0090] Advantageously, the antibody formulations of the present disclosure can be administered to a subject subcutaneously, either by injection or infusion, including by bolus subcutaneous infusion.

[0091] In certain embodiments, the present disclosure relates to formulations of anti-CD38 antibodies comprising a high concentration of antibody, a pH of 5.5 to 7.0, and a viscosity of at most 25 mPa·s at 20°C.

[0092] In certain embodiments, the present disclosure relates to a formulation comprising at least 100 mg / mL of an anti-CD38 antibody, a viscosity-reducing agent, a stabilizer, a buffer, and a surfactant, wherein the formulation has a pH of 5.5 to 7.0 and a viscosity of at most 25 mPa·s at 20°C.

[0093] In some embodiments, a formulation of an antibody comprising at least 100 mg / mL of an anti-CD38 antibody, The anti-CD38 antibody comprises a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, each having the amino acid sequences set forth in SEQ ID NOS: 1 to 3, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, each having the amino acid sequences set forth in SEQ ID NOS: 4 to 6, The formulation comprises a viscosity-reducing agent, a stabilizer, a buffer and a surfactant; Formulations are provided, wherein the formulations have a pH of 5.5 to 7.0 and a viscosity of at most 25 mPa·s at 20°C.

[0094] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody.

[0095] In certain embodiments, the formulation comprises 140 mg / mL of anti-CD38 antibody.

[0096] In one particular embodiment, the viscosity-reducing agent is 90-150 mM Arg-Cl.

[0097] In one particular embodiment, the viscosity-reducing agent is 90-130 mM Arg-Cl.

[0098] In one particular embodiment, the viscosity-reducing agent is 110 mM Arg-Cl.

[0099] In certain embodiments, the stabilizer is sucrose.

[0100] In certain embodiments, the aqueous formulation comprises 125-155 mg / mL of anti-CD38 antibody, 90-150 mM Arg-Cl, and sucrose.

[0101] In certain embodiments, the formulation comprises 140 mg / mL anti-CD38 antibody, 90-150 mM Arg-Cl, and sucrose.

[0102] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 90-130 mM Arg-Cl, and sucrose.

[0103] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 110 mM Arg-Cl, and sucrose.

[0104] In certain embodiments, the formulation comprises 140 mg / mL anti-CD38 antibody, 90-130 mM Arg-Cl, and sucrose.

[0105] In certain embodiments, the formulation comprises 140 mg / mL of anti-CD38 antibody, 110 mM Arg-Cl, and sucrose.

[0106] In certain embodiments, the formulation comprises 2% (w / v) sucrose.

[0107] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 90-150 mM Arg-Cl, and 2% (w / v) sucrose.

[0108] In certain embodiments, the formulation comprises 140 mg / mL anti-CD38 antibody, 90-130 mM Arg-Cl, and 2% (w / v) sucrose.

[0109] In certain embodiments, the formulation comprises 140 mg / mL anti-CD38 antibody, 110 mM Arg-Cl, and 2% (w / v) sucrose.

[0110] In certain embodiments, the formulation comprises a surfactant.

[0111] In one particular embodiment, the surfactant is poloxamer 188.

[0112] In one particular embodiment, the viscosity-reducing agent is 90-150 mM Arg-Cl, the stabilizer is sucrose, and the surfactant is poloxamer 188.

[0113] In one particular embodiment, the viscosity-reducing agent is 90-150 mM Arg-Cl, the stabilizer is 2 (w / v)% sucrose, and the surfactant is poloxamer 188.

[0114] In certain embodiments, the solution comprises 125-155 mg / mL of anti-CD38 antibody, 90-150 mM Arg-Cl, 2% (w / v) sucrose, and poloxamer 188.

[0115] In one particular embodiment, the solution comprises 140 mg / mL anti-CD38 antibody, 90-150 mM Arg-Cl, 2 (w / v)% sucrose, and poloxamer 188.

[0116] In one particular embodiment, the viscosity-reducing agent is 90-130 mM Arg-Cl, the stabilizer is sucrose, and the surfactant is poloxamer 188.

[0117] In one particular embodiment, the viscosity-reducing agent is 110 mM Arg-Cl, the stabilizer is sucrose, and the surfactant is poloxamer 188.

[0118] In one particular embodiment, the viscosity-reducing agent is 90-125 mM Arg-Cl, the stabilizer is 2 (w / v)% sucrose, and the surfactant is poloxamer 188.

[0119] In one particular embodiment, the viscosity-reducing agent is 110 mM Arg-Cl, the stabilizer is 2% (w / v) sucrose, and the surfactant is poloxamer 188.

[0120] In certain embodiments, the solution comprises 125-155 mg / mL of anti-CD38 antibody, 90-125 mM Arg-Cl, 2% (w / v) sucrose, and poloxamer 188.

[0121] In certain embodiments, the solution comprises 125-155 mg / mL of anti-CD38 antibody, 110 mM Arg-Cl, 2 (w / v)% sucrose, and poloxamer 188.

[0122] In certain embodiments, the solution comprises 140 mg / mL anti-CD38 antibody, 90-125 mM Arg-Cl, 2 (w / v)% sucrose, and poloxamer 188.

[0123] In one particular embodiment, the solution comprises 140 mg / mL anti-CD38 antibody, 110 mM Arg-Cl, 2% (w / v) sucrose, and poloxamer 188.

[0124] In certain embodiments, the formulation comprises 0.4% (w / v) poloxamer 188.

[0125] In one particular embodiment, the viscosity-reducing agent is 90-150 mM Arg-Cl, the stabilizer is sucrose, and the surfactant is 0.4 (w / v)% poloxamer 188.

[0126] In one particular embodiment, the viscosity-reducing agent is 90-150 mM Arg-Cl, the stabilizer is 2 (w / v)% sucrose, and the surfactant is 0.4 (w / v)% poloxamer 188.

[0127] In certain embodiments, the solution comprises 125-155 mg / mL of anti-CD38 antibody, 90-150 mM Arg-Cl, 2 (w / v)% sucrose, and 0.4 (w / v)% poloxamer 188.

[0128] In one particular embodiment, the solution comprises 140 mg / mL anti-CD38 antibody, 90-150 mM Arg-Cl, 2 (w / v)% sucrose, and 0.4 (w / v)% poloxamer 188.

[0129] In one particular embodiment, the viscosity-reducing agent is 90-125 mM Arg-Cl, the stabilizer is sucrose, and the surfactant is 0.4 (w / v)% poloxamer 188.

[0130] In one particular embodiment, the viscosity-reducing agent is 110 mM Arg-Cl, the stabilizer is sucrose, and the surfactant is 0.4% (w / v) poloxamer 188.

[0131] In one particular embodiment, the viscosity-reducing agent is 90-125 mM Arg-Cl, the stabilizer is 2 (w / v)% sucrose, and the surfactant is 0.4 (w / v)% poloxamer 188.

[0132] In one particular embodiment, the viscosity-reducing agent is 110 mM Arg-Cl, the stabilizer is 2 (w / v)% sucrose, and the surfactant is 0.4 (w / v)% poloxamer 188.

[0133] In certain embodiments, the solution comprises 125-155 mg / mL of anti-CD38 antibody, 90-125 mM Arg-Cl, 2 (w / v)% sucrose, and 0.4 (w / v)% poloxamer 188.

[0134] In certain embodiments, the solution comprises 125-155 mg / mL of anti-CD38 antibody, 110 mM Arg-Cl, 2 (w / v)% sucrose, and 0.4 (w / v)% poloxamer 188.

[0135] In one particular embodiment, the solution comprises 140 mg / mL anti-CD38 antibody, 90-125 mM Arg-Cl, 2 (w / v)% sucrose, and 0.4 (w / v)% poloxamer 188.

[0136] In certain embodiments, the solution comprises 140 mg / mL anti-CD38 antibody, 110 mM Arg-Cl, 2 (w / v)% sucrose, and 0.4 (w / v)% poloxamer 188.

[0137] In certain embodiments, the formulation comprises a buffering agent.

[0138] In certain embodiments, the buffering agent is histidine.

[0139] In one particular embodiment, the viscosity-reducing agent is 90-150 mM Arg-Cl, the stabilizer is sucrose, the surfactant is poloxamer 188, and the buffering agent is histidine.

[0140] In certain embodiments, the viscosity-reducing agent is 90-150 mM Arg-Cl, the stabilizer is 2 (w / v)% sucrose, the surfactant is poloxamer 188, and the buffering agent is histidine.

[0141] In certain embodiments, the viscosity-reducing agent is 90-150 mM Arg-Cl, the stabilizer is 2% (w / v) sucrose, the surfactant is 0.4% (w / v) poloxamer 188, and the buffering agent is histidine.

[0142] In certain embodiments, the solution comprises 125-155 mg / mL of anti-CD38 antibody, 90-150 mM Arg-Cl, 2 (w / v)% sucrose, 0.4 (w / v)% poloxamer 188, and histidine.

[0143] In one particular embodiment, the solution comprises 140 mg / mL anti-CD38 antibody, 90-150 mM Arg-Cl, 2 (w / v)% sucrose, 0.4 (w / v)% poloxamer 188, and histidine.

[0144] In one particular embodiment, the viscosity-reducing agent is 90-125 mM Arg-Cl, the stabilizer is sucrose, the surfactant is poloxamer 188, and the buffering agent is histidine.

[0145] In one particular embodiment, the viscosity-reducing agent is 110 mM Arg-Cl, the stabilizer is sucrose, the surfactant is poloxamer 188, and the buffering agent is histidine.

[0146] In certain embodiments, the viscosity-reducing agent is 90-125 mM Arg-Cl, the stabilizer is 2 (w / v)% sucrose, the surfactant is poloxamer 188, and the buffering agent is histidine.

[0147] In one particular embodiment, the viscosity-reducing agent is 110 mM Arg-Cl, the stabilizer is 2% (w / v) sucrose, the surfactant is poloxamer 188, and the buffering agent is histidine.

[0148] In certain embodiments, the viscosity-reducing agent is 90-125 mM Arg-Cl, the stabilizer is 2% (w / v) sucrose, the surfactant is 0.4% (w / v) poloxamer 188, and the buffering agent is histidine.

[0149] In one particular embodiment, the viscosity-reducing agent is 110 mM Arg-Cl, the stabilizer is 2% (w / v) sucrose, the surfactant is 0.4% (w / v) poloxamer 188, and the buffering agent is histidine.

[0150] In certain embodiments, the solution comprises 125-155 mg / mL of anti-CD38 antibody, 90-125 mM Arg-Cl, 2 (w / v)% sucrose, 0.4 (w / v)% poloxamer 188, and histidine.

[0151] In certain embodiments, the solution contains 125-155 mg / mL of anti-CD38 antibody, 110 mM Arg-Cl, 2 (w / v)% sucrose, 0.4 (w / v)% poloxamer 188, and histidine.

[0152] In certain embodiments, the solution comprises 140 mg / mL anti-CD38 antibody, 90-125 mM Arg-Cl, 2 (w / v)% sucrose, 0.4 (w / v)% poloxamer 188, and histidine.

[0153] In certain embodiments, the solution comprises 140 mg / mL anti-CD38 antibody, 110 mM Arg-Cl, 2 (w / v)% sucrose, 0.4 (w / v)% poloxamer 188, and histidine.

[0154] In one particular embodiment, the buffering agent is 9 mM histidine.

[0155] In one particular embodiment, the viscosity-reducing agent is 90-150 mM Arg-Cl, the stabilizer is sucrose, the surfactant is poloxamer 188, and the buffering agent is 9 mM histidine.

[0156] In certain embodiments, the viscosity-reducing agent is 90-150 mM Arg-Cl, and the stabilizer The agent is 2% (w / v) sucrose, the surfactant is poloxamer 188, and the buffer is 9 mM histidine.

[0157] In one particular embodiment, the viscosity-reducing agent is 90-150 mM Arg-Cl, the stabilizer is 2% (w / v) sucrose, the surfactant is 0.4% (w / v) poloxamer 188, and the buffering agent is 9 mM histidine.

[0158] In certain embodiments, the solution contains 125-155 mg / mL of anti-CD38 antibody, 90-150 mM Arg-Cl, 2 (w / v)% sucrose, 0.4 (w / v)% poloxamer 188, and 9 mM histidine.

[0159] In one particular embodiment, the solution contains 140 mg / mL anti-CD38 antibody, 90-150 mM Arg-Cl, 2 (w / v)% sucrose, 0.4 (w / v)% poloxamer 188, and 9 mM histidine.

[0160] In one particular embodiment, the viscosity-reducing agent is 90-125 mM Arg-Cl, the stabilizer is sucrose, the surfactant is poloxamer 188, and the buffering agent is 9 mM histidine.

[0161] In one particular embodiment, the viscosity-reducing agent is 110 mM Arg-Cl, the stabilizer is sucrose, the surfactant is poloxamer 188, and the buffering agent is 9 mM histidine.

[0162] In one particular embodiment, the viscosity-reducing agent is 90-125 mM Arg-Cl, the stabilizer is 2% (w / v) sucrose, the surfactant is poloxamer 188, and the buffering agent is 9 mM histidine.

[0163] In one particular embodiment, the viscosity-reducing agent is 110 mM Arg-Cl, the stabilizer is 2% (w / v) sucrose, the surfactant is poloxamer 188, and the buffering agent is 9 mM histidine.

[0164] In one particular embodiment, the viscosity-reducing agent is 90-125 mM Arg-Cl, the stabilizer is 2% (w / v) sucrose, the surfactant is 0.4% (w / v) poloxamer 188, and the buffering agent is 9 mM histidine.

[0165] In one particular embodiment, the viscosity-reducing agent is 110 mM Arg-Cl, the stabilizer is 2% (w / v) sucrose, the surfactant is 0.4% (w / v) poloxamer 188, and the buffering agent is 9 mM histidine.

[0166] In certain embodiments, the solution comprises 125-155 mg / mL of anti-CD38 antibody, 90-125 mM Arg-Cl, 2 (w / v)% sucrose, 0.4 (w / v)% poloxamer 188, and 9 mM histidine.

[0167] In certain embodiments, the solution contains 125-155 mg / mL of anti-CD38 antibody, 110 mM Arg-Cl, 2 (w / v)% sucrose, 0.4 (w / v)% poloxamer 188, and 9 mM histidine.

[0168] In one particular embodiment, the solution contains 140 mg / mL anti-CD38 antibody, 90-125 mM Arg-Cl, 2 (w / v)% sucrose, 0.4 (w / v)% poloxamer 188, and 9 mM histidine.

[0169] In one particular embodiment, the solution comprises 140 mg / mL anti-CD38 antibody, 110 mM Arg-Cl, 2 (w / v)% sucrose, 0.4 (w / v)% poloxamer 188, and 9 mM histidine.

[0170] In certain embodiments, the viscosity-reducing agent is 90-150 mM Lys-Ac.

[0171] In one particular embodiment, the viscosity-reducing agent is 125 mM Lys-Ac.

[0172] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 90-150 mM Lys-Ac and sucrose.

[0173] In certain embodiments, the formulation comprises 140 mg / mL anti-CD38 antibody, 90-150 mM Lys-Ac and sucrose.

[0174] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 125 mM Lys-Ac and sucrose.

[0175] In certain embodiments, the formulation comprises 140 mg / mL of anti-CD38 antibody, 125 mM Lys-Ac and sucrose.

[0176] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 90-150 mM Lys-Ac, and 2% (w / v) sucrose.

[0177] In certain embodiments, the formulation comprises 140 mg / mL anti-CD38 antibody, 90-150 mM Lys-Ac, and 2% (w / v) sucrose.

[0178] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 125 mM Lys-Ac, and 2% (w / v) sucrose.

[0179] In certain embodiments, the formulation comprises 140 mg / mL of anti-CD38 antibody, 125 mM Lys-Ac, and 2% (w / v) sucrose.

[0180] In certain embodiments, the formulation comprises a surfactant.

[0181] In certain embodiments, the surfactant is polysorbate 80.

[0182] In certain embodiments, the viscosity-reducing agent is 90-150 mM Lys-Ac, the stabilizer is sucrose, and the surfactant is polysorbate 80.

[0183] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 90-150 mM Lys-Ac, sucrose, and polysorbate 80.

[0184] In certain embodiments, the formulation comprises 140 mg / mL anti-CD38 antibody, 90-150 mM Lys-Ac, sucrose, and polysorbate 80.

[0185] In certain embodiments, the viscosity-reducing agent is 90-150 mM Lys-Ac, the stabilizer is 2% (w / v) sucrose, and the surfactant is polysorbate 80.

[0186] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 90- Contains 150 mM Lys-Ac, 2 (w / v)% sucrose and polysorbate 80.

[0187] In certain embodiments, the formulation comprises 140 mg / mL anti-CD38 antibody, 90-150 mM Lys-Ac, 2% (w / v) sucrose, and polysorbate 80.

[0188] In one particular embodiment, the viscosity-reducing agent is 125 mM Lys-Ac, the stabilizer is sucrose, and the surfactant is polysorbate 80.

[0189] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 125 mM Lys-Ac, sucrose, and polysorbate 80.

[0190] In certain embodiments, the formulation comprises 140 mg / mL of anti-CD38 antibody, 125 mM Lys-Ac, sucrose, and polysorbate 80.

[0191] In certain embodiments, the viscosity-reducing agent is 125 mM Lys-Ac, the stabilizer is 2% (w / v) sucrose, and the surfactant is polysorbate 80.

[0192] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 125 mM Lys-Ac, 2% (w / v) sucrose, and polysorbate 80.

[0193] In certain embodiments, the formulation comprises 140 mg / mL of anti-CD38 antibody, 125 mM Lys-Ac, 2% (w / v) sucrose and polysorbate 80.

[0194] In one particular embodiment, the surfactant is 0.04% (w / v) polysorbate 80.

[0195] In certain embodiments, the viscosity-reducing agent is 90-150 mM Lys-Ac, the stabilizer is sucrose, and the surfactant is 0.04 (w / v)% polysorbate 80.

[0196] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 90-150 mM Lys-Ac, sucrose, and 0.04% (w / v) polysorbate 80.

[0197] In certain embodiments, the formulation comprises 140 mg / mL anti-CD38 antibody, 90-150 mM Lys-Ac, sucrose, and 0.04% (w / v) polysorbate 80.

[0198] In certain embodiments, the viscosity-reducing agent is 90-150 mM Lys-Ac, the stabilizer is 2 (w / v)% sucrose, and the surfactant is 0.04 (w / v)% polysorbate 80.

[0199] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 90-150 mM Lys-Ac, 2% (w / v) sucrose, and 0.04% (w / v) polysorbate 80.

[0200] In certain embodiments, the formulation comprises 140 mg / mL anti-CD38 antibody, 90-150 mM Lys-Ac, 2% (w / v) sucrose, and 0.04% (w / v) polysorbate 80.

[0201] In one particular embodiment, the viscosity-reducing agent is 125 mM Lys-Ac, the stabilizer is sucrose, and the surfactant is 0.04% (w / v) polysorbate 80.

[0202] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 125 mM Lys-Ac, sucrose, and 0.04% (w / v) polysorbate 80.

[0203] In certain embodiments, the formulation comprises 140 mg / mL of anti-CD38 antibody, 125 mM Lys-Ac, sucrose, and 0.04% (w / v) polysorbate 80.

[0204] In certain embodiments, the viscosity-reducing agent is 125 mM Lys-Ac, the stabilizer is 2% (w / v) sucrose, and the surfactant is 0.04% (w / v) polysorbate 80.

[0205] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody, 125 mM Lys-Ac, 2% (w / v) sucrose, and 0.04% (w / v) polysorbate 80.

[0206] In certain embodiments, the formulation comprises 140 mg / mL anti-CD38 antibody, 125 mM Lys-Ac, 2% (w / v) sucrose, and 0.04% (w / v) polysorbate 80.

[0207] In certain embodiments, the pH of the formulation is between 5.9 and 7.0.

[0208] In certain embodiments, the pH of the formulation is between 5.9 and 6.5.

[0209] In certain embodiments, the pH of the formulation is 6.2.

[0210] In some embodiments, the formulation comprises 140 mg / mL of an anti-CD38 antibody, the anti-CD38 antibody comprising a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, comprising the amino acid sequences set forth in SEQ ID NOs: 1 to 3, respectively, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, comprising the amino acid sequences set forth in SEQ ID NOs: 4 to 6, respectively; The formulation contained 9 mM histidine, 110 mM Arg-Cl, 2 (w / v)% sucrose and 0.4 (w / v)% poloxamer 188; The formulation has a pH of 6.2-6.3 and a viscosity of at most 14 mPa·s at 20°C.

[0211] One embodiment of the present disclosure is a formulation comprising 140 mg / mL of an anti-CD38 antibody, wherein the anti-CD38 antibody comprises a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences set forth in SEQ ID NOS: 1 to 3, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences set forth in SEQ ID NOS: 4 to 6, The formulation contained 125 mM Lys-Ac, 2 (w / v)% sucrose and 0.04 (w / v)% polysorbate 80; The formulation has a pH of 6.2 and a viscosity of at most 14 mPa·s at 20°C.

[0212] According to each of the foregoing aspects and embodiments, in certain embodiments, the VH of the anti-CD38 antibody comprises the amino acid sequence set forth in SEQ ID NO:7.

[0213] According to each of the foregoing aspects and embodiments, in certain embodiments, the VL of the anti-CD38 antibody comprises the amino acid sequence set forth in SEQ ID NO:8.

[0214] According to each of the foregoing aspects and embodiments, in certain embodiments, the VH of the anti-CD38 antibody comprises the amino acid sequence set forth in SEQ ID NO:7, and the VL of the anti-CD38 antibody comprises the amino acid sequence set forth in SEQ ID NO:8.

[0215] According to each of the foregoing aspects and embodiments, in certain embodiments, the anti-CD38 antibody is isatuximab.

[0216] In accordance with each of the foregoing aspects and embodiments, the formulation further comprises water, e.g., water for injection (WFI), in an amount sufficient to achieve the specified concentrations of the other ingredients.

[0217] According to each of the above aspects and embodiments, in certain embodiments, the formulation is suitable for subcutaneous administration.For example, the formulation can be sterile.In certain embodiments, the components of the formulation can be combined to form a solution, and then the solution can be sterile filtered to provide a sterile formulation.

[0218] In certain embodiments, the finished formulation is substantially free of dissolved oxygen. For example, the formulation can be equilibrated with nitrogen gas and then sealed under a nitrogen atmosphere.

[0219] Further according to each of the foregoing aspects and embodiments, in certain embodiments, the formulation may further comprise at least one additional excipient or ingredient for enhanced stability, such as a preservative.

[0220] Packaged pharmaceutical products Some aspects of the present disclosure relate to a packaged pharmaceutical product, comprising a sterile container containing a single dose of the formulation of the present disclosure.Suitable sterile containers include, but are not limited to, vials, ampoules, bottles, bags, pouches, pre-filled syringes, syringe drivers, infusion pumps, and the containers adapted for use with syringe drivers and / or infusion pumps.Suitable containers include disposable containers and multi-use containers.In certain embodiments, the container is a disposable container, for example, a vial that contains an amount of antibody corresponding to a single dose.

[0221] Syringe driver as used herein refers to a mechanical or pneumatic device that is constructed and arranged to engage the plunger of a syringe and drive it axially forward and / or backward so that the contents of the syringe are delivered at a desired speed.Syringe drivers are known in the art, and include, but are not limited to, the devices disclosed in U.S. Patent Nos. 5,064,413; 5,449,345; 5,954,695; 6,428,509; 6,645,177; 7,195,610; 8,231,576; and 8,814,830, the entire contents of which are incorporated herein by reference.

[0222] Infusion pumps are well known in the art and include, for example, the Baxter Colleague CXE volumetric infusion pump and the Cane Crono pump.

[0223] Certain aspects of the present disclosure relate to a device comprising a therapeutically effective amount of a formulation of the present disclosure. In certain embodiments, the device may be, for example, a syringe containing the formulation, a syringe driver, and an infusion pump. In certain embodiments, the syringe is a pre-filled syringe. do.

[0224] In some embodiments, the antibody formulations of the present disclosure are provided in a fixed-dose format. Such formulations can be provided, for example, in or as vials or ampoules. For example, in some embodiments, the antibody formulations of the present disclosure are provided in a volume of about 10 mL to about 20 mL. In some embodiments, the antibody formulations of the present disclosure are provided in a volume of about 10 mL to about 15 mL. In some embodiments, the antibody formulations of the present disclosure are provided in a volume of about 10 mL to about 12.5 mL. For example, in one embodiment of a formulation comprising 140 mg / mL of antibody, a vial containing 10 mL of such a formulation contains 1400 mg of antibody.

[0225] Treatment method The formulations of the present disclosure contain CD38 + The present invention can be used in methods for treating diseases or conditions characterized by the presence or activity of CD38 cells. Such diseases or conditions include, but are not limited to, + These may include hematological malignancies, autoimmune diseases or conditions, inflammatory diseases or conditions, and LPS- or sepsis-induced kidney damage or dysfunction. The method generally involves administering to a subject in need thereof an effective amount of a formulated antibody provided herein, optionally by subcutaneous injection or infusion, optionally with a large volume (e.g., 10 mL or more) of subcutaneous injection. In certain embodiments, the subject is a human.

[0226] Certain aspects of the present disclosure provide for the administration of CD38 in a human subject in need thereof. + 1. A method of treating a hematological malignancy, comprising administering to said human subject an effective amount of a formulation comprising at least 100 mg / mL of an anti-CD38 antibody; The anti-CD38 antibody comprises a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, each having the amino acid sequences set forth in SEQ ID NOS: 1 to 3, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, each having the amino acid sequences set forth in SEQ ID NOS: 4 to 6, The formulation comprises a viscosity-reducing agent, a stabilizer, a buffer and a surfactant; The formulation has a pH of 5.7 to 7.0 and a viscosity of at most 25 mPa·s at 20°C, and is administered subcutaneously.

[0227] In one particular embodiment, the viscosity-reducing agent is 90-150 mM Arg-Cl.

[0228] In one particular embodiment, the viscosity-reducing agent is 90-130 mM Arg-Cl.

[0229] In one particular embodiment, the viscosity-reducing agent is 110 mM Arg-Cl.

[0230] In one particular embodiment, the surfactant is poloxamer 188.

[0231] In one particular embodiment, the surfactant is 0.4% (w / v) poloxamer 188.

[0232] In certain embodiments, the buffering agent is histidine.

[0233] In one particular embodiment, the buffering agent is 9 mM histidine.

[0234] In certain embodiments, the viscosity-reducing agent is 90-150 mM Lys-Ac.

[0235] In one particular embodiment, the viscosity-reducing agent is 125 mM Lys-Ac.

[0236] In certain embodiments, the surfactant is polysorbate 80.

[0237] In one particular embodiment, the surfactant is 0.04% (w / v) polysorbate 80.

[0238] In certain embodiments, the formulation comprises 125-155 mg / mL of anti-CD38 antibody.

[0239] In certain embodiments, the formulation comprises 140 mg / mL of anti-CD38 antibody.

[0240] In certain embodiments, the stabilizer is sucrose.

[0241] In certain embodiments, the stabilizer is 2% (w / v) sucrose.

[0242] In certain embodiments, the pH of the formulation is between 5.9 and 7.0.

[0243] In certain embodiments, the pH of the formulation is between 5.9 and 6.5.

[0244] In certain embodiments, the pH of the formulation is 6.2.

[0245] In certain embodiments, the pH of the formulation is 6.3.

[0246] Certain aspects of the present disclosure provide for the administration of CD38 in a human subject in need thereof. + 1. A method of treating a hematological malignancy, comprising administering to said human subject an effective amount of a formulation comprising 140 mg / mL of an anti-CD38 antibody; The anti-CD38 antibody comprises a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, each having the amino acid sequences set forth in SEQ ID NOS: 1 to 3, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, each having the amino acid sequences set forth in SEQ ID NOS: 4 to 6, The formulation contained 9 mM histidine, 110 mM Arg-Cl, 2 (w / v)% sucrose and 0.4 (w / v)% poloxamer 188; The formulation has a pH of 6.2 and a viscosity of at most 14 mPa·s at 20°C, and is administered subcutaneously.

[0247] Certain aspects of the present disclosure provide for the administration of CD38 in a human subject in need thereof. +1. A method of treating a hematological malignancy, comprising administering to said human subject an effective amount of a formulation comprising 140 mg / mL of an anti-CD38 antibody; The anti-CD38 antibody comprises a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, each having the amino acid sequences set forth in SEQ ID NOS: 1 to 3, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, each having the amino acid sequences set forth in SEQ ID NOS: 4 to 6, The formulation contained 125 mM Lys-Ac, 2 (w / v)% sucrose and 0.04 (w / v)% polysorbate 80, The formulation has a pH of 6.2 and a viscosity of at most 14 mPa·s at 20°C, and is administered subcutaneously.

[0248] In certain embodiments according to each of the above methods, administering subcutaneously comprises one or more subcutaneous injections.

[0249] In certain embodiments according to each of the above methods, administering subcutaneously comprises one or more subcutaneous injections.

[0250] In certain embodiments according to each of the above methods, administering subcutaneously comprises one or more bolus subcutaneous injections.

[0251] As used herein, a "large injection" refers to an injection volume of 5 mL or greater. In certain embodiments, a "large injection" refers to an injection volume of about 5-10 mL, about 10-15 mL, about 15-20 mL, about 20-25 mL, or about 25-30 mL. In some embodiments, a "large injection" refers to an injection volume of about 5-10 mL. In some embodiments, a "large injection" refers to an injection volume of about 10-15 mL. In some embodiments, a "large injection" refers to an injection volume of about 15-20 mL. In some embodiments, a "large injection" refers to an injection volume of about 20-25 mL. In some embodiments, a "large injection" refers to an injection volume of about 25-30 mL.

[0252] Surprisingly, high-dose injections of the compositions provided herein suppress CD38 expression in humans. + CD38 including hematologic malignancies + It has been found that the systemic delivery of therapeutically effective amounts of anti-CD38 antibodies, such as isatuximab, to treat diseases or conditions characterized by the presence and / or activity of CD38 cells is effective. Surprisingly, the antibody formulations provided herein demonstrated at least 89% bioavailability when administered subcutaneously to minipigs in the absence of a dispersing agent, such as hyaluronidase. Thus, provided herein are formulations that do not contain a biological dispersing agent. In alternative embodiments, provided herein are formulations that further comprise one or more biological dispersing agents.

[0253] As disclosed in Example 6, following a single intravenous (IV) infusion of isatuximab at 1800 mg per animal over 30 minutes into minipigs, the mean AUC (AUC last After a single subcutaneous (SC) infusion of 1806 mg of isatuximab per animal into minipigs at flow rates of 0.5, 1, or 2 mL / min, the mean AUC (AUC last ) were 326,000, 565,000, and 369,000 h*μg / mL, respectively. Furthermore, the absolute SC bioavailability of isatuximab in minipigs was at least 89% when given at a dose of 1806 mg (140 mg / mL solution) per animal by SC infusion at a flow rate of 0.5–2 mL / min.

[0254] As used herein, a "subject" and / or a "subject in need thereof" refers to a subject who is a patient with a CD38 + have hematologic malignancies or CD38 + An individual suspected of having a hematological malignancy. As used herein, "subject" can also refer to a patient.

[0255] A subject according to the present disclosure may be male or female.

[0256] In some embodiments, the subject has previously been treated with one or more agents or therapies suitable for treating a CD38-expressing hematological malignancy. The previous anti-cancer therapy may be, for example, a corticosteroid (e.g., dexamethasone), a chemotherapeutic agent, a proteasome inhibitor, an immunomodulatory agent, radiation therapy, bone marrow and / or stem cell transplantation, and immunotherapy.

[0257] A "chemotherapeutic agent" is a cytotoxic agent used, for example, to treat hematological malignancies, and includes, without limitation, cytarabine (cytosine arabinoside or ara-C) and anthracycline drugs (such as daunorubicin and / or daunomycin, doxorubicin and liposomal doxorubicin, idarubicin and mitoxantrone), gemtuzumab, clofarabine, cladribine, hydroxyurea, etoposide, amsacrine, FLT3 inhibitors, and demethylating agents (5-azacytidine and decitabine), melphalan, cyclophosphamide, and vincristine.

[0258] Proteasome inhibitors include, for example, bortezomib, carfilzomib, and ixazomib. Immunomodulatory agents include, for example, thalidomide, lenalidomide, and pomalidomide.

[0259] "Radiotherapy" or "radiation" refers to high-energy radiation used to eliminate cancer cells. Radiation therapy may be used prior to bone marrow or peripheral blood stem cell transplantation.

[0260] "Bone marrow and / or stem cell transplant" refers to a cell transplant aimed at restoring stem cells destroyed by high doses of chemotherapy drug(s) and / or radiation therapy. Sources of stem cells include bone marrow, peripheral blood, or umbilical cord blood. Depending on the source of the transplanted stem cells, the procedure can be classified as bone marrow transplant (BMT), peripheral blood stem cell transplant (PBSCT), or umbilical cord blood transplant (UCBT).

[0261] Additionally, bone marrow and / or stem cell transplantation can refer to autologous stem cell transplantation and / or allogeneic transplantation.

[0262] In an "autologous transplant," a subject's own stem cells are removed, frozen, and stored from their bone marrow or peripheral blood while they undergo treatment (high-dose chemotherapy drug(s) and / or radiation). A process called "purging" can be used to attempt to remove any cancer cells in the sample. The stem cells are then reinfused into the subject's blood after treatment.

[0263] An "allogeneic transplant" is a transplant from a matched donor. The advantage of an allogeneic bone marrow transplant is that the transplanted cells from the donor can detect leukemia cells as foreign and establish a new immune system to eliminate them. The disadvantages of an allogeneic transplant are the limitations of the matched donor and the side effects.

[0264] "Immunotherapy" refers to the stimulation of the subject's immune system to attack the malignant tumor cells that cause disease.This can be achieved, for example, by immunizing the subject by administering a cancer vaccine, when the subject's own immune system is trained to recognize tumor cells as targets to be destroyed, or by administering a therapeutic antibody as a drug, when the subject's immune system is mobilized to destroy tumor cells by the therapeutic antibody.

[0265] In the context of the disclosure, the subject has been previously treated for a hematological malignancy, but has relapsed and / or is refractory.

[0266] In some embodiments, the subject has multiple myeloma. In some embodiments, the subject has relapsed and / or refractory multiple myeloma.

[0267] "Relapsed" refers to a disease or condition, such as a hematological malignancy, that has been previously treated but has progressed to require the initiation of further treatment but does not meet the criteria for primary refractory or relapsed and refractory disease.

[0268] "Refractory" refers to a disease or condition that is non-responsive during primary or salvage therapy (fails to achieve a minimal response during therapy or develops progressive disease) or progresses within 60 days of the last therapy.

[0269] Relapsed refractory disease is either non-responsive during salvage therapy (e.g., therapy administered after failure of first-line therapy) or progresses through their current disease course. Disease progression within 60 days of last therapy in patients who achieved at least a minimal response at some point prior to receiving treatment.

[0270] Primary refractory disease is non-responsive disease in patients who have never achieved more than a minimal response to any therapy.

[0271] In some embodiments, the subject has been previously treated with bortezomib and / or lenalidomide.

[0272] In some embodiments, the subject has previously undergone autologous stem cell transplantation (ASCT).

[0273] In some embodiments, the subject relapses within six months of the autologous transplant.

[0274] Dosage and Administration In some embodiments, the formulated antibodies provided herein are administered as a "flat dose" such that the amount of antibody administered to a patient is not adjusted based on body size or weight. In some embodiments, the flat dose administered to a patient comprises 1000-1800 mg of antibody. In some embodiments, the flat dose is 1000 mg. In some embodiments, the flat dose is 1400 mg.

[0275] In some embodiments, formulated antibodies, e.g., fixed-dose antibodies, are administered to a patient in a fixed volume. For example, in some embodiments, the antibody formulations of the present disclosure are administered in a volume of about 10 to about 20 mL. In some embodiments, the antibody formulations of the present disclosure are administered in a volume of about 10 to about 15 mL. In some embodiments, the antibody formulations of the present disclosure are administered in a volume of about 10 mL to about 12.5 mL. In some embodiments, the antibody formulations of the present disclosure are administered in a volume of about 10 mL to about 11 mL. In some embodiments, the antibody formulations of the present disclosure are administered in a volume of about 10 mL to about 10.5 mL.

[0276] In some embodiments, a dose of a formulated antibody provided herein is administered subcutaneously over about 10 to about 60 minutes. In some embodiments, a dose of a formulated antibody is administered subcutaneously over about 20 to about 40 minutes. In some embodiments, a dose of a formulated antibody is administered subcutaneously over about 10 minutes. In some embodiments, a dose of a formulated antibody is administered subcutaneously over about 20 minutes. In some embodiments, a dose of a formulated antibody is administered subcutaneously over about 30 minutes. In some embodiments, a dose of a formulated antibody is administered subcutaneously over about 40 minutes. In some embodiments, a dose of a formulated antibody is administered subcutaneously over about 50 minutes. In some embodiments, a dose of a formulated antibody is administered subcutaneously over about 60 minutes.

[0277] In some embodiments, subcutaneous administration occurs at a certain rate of antibody infusion. For example, the formulation can be administered subcutaneously at a rate suitable to achieve complete delivery of the desired dose in a minimum amount of time without significant leakage or discomfort. Such a rate can range, for example, from about 0.1 mL / min to about 1.5 mL / min. In some embodiments, the infusion rate is 0.8 mL / min. In some embodiments, the rate is 1 mL / min. In some embodiments, the rate is 1.2 mL / min. In some embodiments, the rate is 1.5 mL / min.

[0278] In some embodiments, the initial rate of infusion can be maintained for the entire duration of the infusion, while in other embodiments the rate of infusion can be adjusted up or down or up or down during the infusion period.

[0279] In some embodiments, the anti-CD38 antibody is administered alone. In other embodiments, the anti-CD38 antibody is administered alone. + It is administered together with another agent suitable for treating blood cancer. In some embodiments, the other agent is a corticosteroid (eg, dexamethasone), a chemotherapeutic agent, a proteasome inhibitor, an immunomodulatory agent, or a combination thereof.

[0280] CD38 + When administered together with another drug suitable for treating blood cancer, the anti-CD38 antibody and the other drug(s) can be administered simultaneously or separately (e.g., sequentially over a period of time). The anti-CD38 antibody and the other drug(s) can be administered by the same or different administration routes. When the anti-CD38 antibody and the other drug(s) are administered by the same administration route, they can be administered by the same or different administration sites.

[0281] Corticosteroids such as dexamethasone are used to treat various inflammatory, autoimmune, and allergic conditions. They are also used in cancer treatment, either as direct agents (e.g., in multiple myeloma) or in combination with other agents (e.g., immunomodulators, chemotherapy agents, and proteasome inhibitors). Corticosteroids such as dexamethasone can also be used to counteract side effects (e.g., nausea and inflammation) of chemotherapy agent(s). Corticosteroids such as dexamethasone can also be used as premedications to reduce the potential risk and / or severity of infusion reactions (IR) from antibody infusions. Dexamethasone is usually administered orally.

[0282] Pomalidomide is a thalidomide analog and immunomodulatory drug with multiple cellular effects that inhibit the growth and survival of multiple myeloma cells and block stromal support from the bone marrow microenvironment, which can promote myeloma cell growth. Furthermore, pomalidomide has strong immunomodulatory effects that enhance the immune response to myeloma cells by stimulating natural killer (NK) cells and inhibiting regulatory T cells. Pomalidomide is usually administered orally.

[0283] In the context of this disclosure, a physician can assess disease response and adjust the dosing regimen accordingly.

[0284] In other embodiments, the anti-CD38 antibody is administered together with one or more biological dispersants. When administered together with a biological dispersant, the anti-CD38 antibody and the other agent(s) can be administered simultaneously or separately (e.g., sequentially over a period of time). The anti-CD38 antibody and the other agent(s) can be administered by the same or different administration routes. When the anti-CD38 antibody and the other agent(s) are administered by the same administration route, they can be administered by the same or different administration sites.

[0285] Prior to administration of the antibody, the subject is premedicated to reduce the risk and / or severity of infusion reactions (IR), which are commonly observed with the administration of monoclonal antibodies. The premedication may include, for example, montelukast, acetaminophen, ranitidine, diphenyldramamine, dexamethasone, or a combination thereof. In some embodiments, if the subject does not experience IR after four consecutive doses of the antibody described herein, the premedication may be discontinued.

[0286] "Disease response" can be determined according to standard criteria for hematological malignancies and staging. +Methods for assessing disease response in hematological malignancies are known to those skilled in the art. For example, methods for assessing disease response include the Eastern Cooperative Oncology Group (ECOG) performance status and the International Myeloma Working Group Response Criteria (Oken et al., J. Clin. Oncol., 2006, 111:111-112, respectively). 1982;5(6):649-655 and Kumar et al., Lancet Oncol. 2016;17(8):328-346). Methods for assessing disease response can also include quantification of disease markers, bone marrow biopsy and / or aspirate, radiological imaging of plasmacytomas, skeletal surveys, M-protein quantification (serum and / or 24-hour urine) and serum free light chain levels or urine light chain levels, serum β2-microglobulin, lymph node biopsy, radiological oncology evaluation (by X-ray, computed tomography (CT) scan, PET scan, or magnetic resonance imaging (MRI)), and blood counts, including blast counts. This list of assessment methods should be understood to be non-limiting.

[0287] Based on the results obtained from the assessment of disease response, disease response can then be stratified by standard criteria for the underlying disease and classified as complete response or complete remission (CR), partial response (PR), stable disease (SD), or progressive disease (PD).

[0288] "Markers" used in the context of response assessment can include serum and / or plasma markers, such as C-reactive protein (CRP), tumor necrosis factor alpha (TNF-α), IL-6, IL-1β, or IFN. Markers can also include cell surface markers such as CD38.

[0289] Techniques for assessing disease response in subjects with multiple myeloma include, for example, bone marrow biopsy and / or aspiration, radiological imaging of plasmacytomas, skeletal survey, M protein quantification, and measurement of serum β2-microglobulin.

[0290] Disease response assessment can further include receptor density and receptor occupancy on circulating tumor cells (peripheral blood), receptor density and receptor occupancy on blasts and plasma cells in the bone marrow, and levels of human anti-drug antibodies (ADA).

[0291] The entire contents of all patents and published patent applications cited in this disclosure are hereby incorporated by reference.

[0292] The present disclosure will be further understood with reference to the following non-limiting examples, which are provided below for illustrative purposes and to describe certain embodiments of the present disclosure. The claims are not limited in any way by the examples provided herein. [Example]

[0293] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the methods and compositions of the present invention, and are not intended to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.). [Example]

[0294] Initial screening Early formulation development activities included screening buffer-pH systems, thermal stabilizers, surfactants, and viscosity-reducing agents to identify excipients and combinations thereof that are compatible with isatuximab and enhance its stability while maintaining an osmolality and viscosity suitable for subcutaneous injection.

[0295] Buffer and pH system selection To identify the buffer and pH system, the stability of isatuximab was evaluated in 16 different buffer-pH systems. The buffer-pH systems (Table 1) were selected to demonstrate their stability in the desired pH range. These were tested based on their buffering capacity.

[0296] [Table 1]

[0297] Buffer-pH systems were evaluated for their effect on isatuximab aggregation with respect to the formation of visible and subvisible particles and soluble aggregates (high molecular weight species, HMW) after shaking and heat stress in a liquid formulation of isatuximab at a concentration of 5 mg / mL.

[0298] As shown in Table 2, aggregation of isatuximab into visible particles was found to be dependent on pH and buffer system. Histidine buffer systems with a pH range of 5.5 to 6.5 showed the highest stability under shaking stress (showing fewer visible particles after stress), while citrate buffer systems within pH 5.0 to 7.0 and citrate buffers at pH 5.5, 6.5, and 7.0 showed the highest stability after heat stress (showing fewer visible particles after 2 weeks of stress). Interestingly, phosphate buffer systems with a pH range of 6.5 to 7.4 showed the highest thermal stability after 1 week of heat stress, but had some visible particles after 2 weeks of heat stress, indicating the lowest stability of isatuximab under shaking stress among all buffer systems tested.

[0299] [Table 2]

[0300] The number of subvisible particles ≥10 μm and ≥25 μm after shaking and heat stress was measured by darkness (LO). As shown in Figure 1, under shaking and heat stress, phosphate buffer showed the highest level of subvisible particles. The histidine and acetate buffer systems showed the lowest level of subvisible particles, indicating greater stability of isatuximab.

[0301] Soluble aggregates (HMW) were observed by size-exclusion chromatography (SE-HPLC) after heat stress in citrate, histidine, phosphate, succinate, and acetate buffers at various pHs. As shown in Figure 2, citrate, phosphate, and succinate buffer systems showed a higher increase in soluble aggregates, and a general trend was observed indicating that the higher the pH, the higher the soluble aggregate (HMW) content. This was particularly significant at pHs above 7.0. Shaking stress did not appear to have any effect on soluble aggregates.

[0302] The effect of pH on isatuximab stability was further studied in histidine buffer in the presence of sucrose and polysorbate 80. Isatuximab at a concentration of 5 mg / mL in the formulations shown in Table 3 was incubated at 40°C for 1 month and the HMW was measured by SE-HPLC.

[0303] [Table 3]

[0304] HMW was observed by SE-HPLC. As shown in Figure 3, there were fewer soluble aggregates (HMW) after 1 month of heat stress at 40°C in histidine buffer at pH 6.0 compared to pH 6.5.

[0305] The levels of the acidic isoform of isatuximab were measured by WCX (weekly cation exchange) after incubation of 5 mg / mL isatuximab in the formulations shown in Table 3 for 1 month at 40° C. As shown in Figure 4, the histidine formulation at pH 6.0 showed a smaller increase in the levels of the acidic form of isatuximab compared to pH 6.5 after 1 month at 40° C.

[0306] These results indicated that pH and buffer system affected the stability of isatuximab, with a pH around pH 6.0 being more stable than pH 6.5.

[0307] Selection of viscosity-reducing excipients Various formulation conditions with viscosity-reducing excipients were tested to determine whether a high viscosity concentration of less than 25 cP at 20° C. could be developed to enable subcutaneous delivery of isatuximab.

[0308] The antibodies were concentrated and formulated into test formulations, the protein concentrations of the resulting formulations were confirmed spectroscopically with a SoloVPE instrument, and the pH of the final solutions was measured.

[0309] [Table 4-1] [Table 4-2] [Table 4-3]

[0310] The viscosity of all samples was measured at 20 °C with a RheoSense Initium instrument using its automated method, which automatically determines the appropriate shear rate for the instrument to operate in the desired range of pressure for the sensor. Because shear rates did not exceed 10,000 s, non-Newtonian effects such as shear thinning are considered negligible.

[0311] Figure 5 shows the viscosity (cP) of a 200 mg / mL solution of isatuximab at pH 6.0 in the presence of 0, 50, 100, and 200 mM L-arginine-Cl. Preliminary measurements showed that arginine-Cl was an effective viscosity-reducing excipient in a concentration-dependent manner; increasing arginine-Cl concentrations correlated with decreased viscosity.

[0312] Over 40 different formulations with various pH and viscosity-reducing agents for various concentrations of isatuximab were tested. The results are shown in Table 4.

[0313] The effect of arginine-Cl concentration on viscosity is shown in Figures 6A and 6B for two concentrations of isatuximab, 150 mg / mL (Figure 6A) and 180 mg / mL (Figure 6B), in the presence of 10 mM histidine and 2% sucrose.

[0314] In addition to the concentration of arginine-Cl, a significant effect of pH on the viscosity of isatuximab was observed. Figures 7A and 7B show the viscosity as a function of pH for 150 mM arginine-Cl (Figure 7A) and 200 mM arginine-Cl (Figure 7B).

[0315] Surprisingly, the results shown in Figures 7A and 7B indicate an inverse relationship between viscosity and pH, i.e., an increase in viscosity is associated with a decrease in pH. There was a sharp increase in viscosity at pH ≤ 5.7. This inverse relationship is the opposite of what is typically expected and observed in protein solutions. Furthermore, this unexpected effect was further amplified with increasing antibody concentration (Figure 8).

[0316] It was shown that an arginine-Cl concentration of at least 100 mM reduced the viscosity of 150 mg / mL isatuximab to less than 20 cp and that of 180 mg / mL isatuximab to less than 40 cp. Furthermore, pH was shown to have a significant effect on viscosity, as demonstrated by the sharp increase in viscosity at pH below 5.7. The effect of pH on the stability of the mab had to be considered. As demonstrated in Figures 3 and 4, histidine buffer systems with pH values ​​close to 6.0 resulted in less HMW compared to the buffer system at pH 6.5 after 1 month of heat stress. [Example]

[0317] viscosity research With the results of the study described in Example 1 at hand, and with the aim of arriving at a formulation containing high concentrations of isatuximab (e.g., at least 100 mg / mL) potentially suitable for subcutaneous administration, several formulations were prepared and studied in more detail. Different parameters, such as protein concentration, pH, viscosity-reducing agent concentration, and temperature, could affect the viscosity of the protein in solution. Several experiments were performed to evaluate the influence of protein concentration, pH, and arginine concentration on the formulation viscosity. Table 5 shows the parameters tested and the value ranges. Considering that the drug product is stored in a frozen state (5±3°C), the influence of temperature was also examined.

[0318] [Table 5]

[0319] The target concentration was set at 140 mg / mL to achieve a viscosity of less than 25 cP at 20° C., including when the mAb solution exhibits variations in antibody or viscosity-reducing agent concentration or pH that are inherent to the manufacturing process. In fact, such variations between the actual and target composition of each excipient are commonly observed in UF / DF during the blending process due to the Donnan effect or due to inaccuracies in excipient metering or other manufacturing steps (e.g., filtration) that may affect the excipient levels in the final drug product.

[0320] Solutions of mAbs at high concentrations tend to exhibit high viscosity. To determine whether a highly concentrated liquid formulation of isatuximab with a viscosity less than 25 mPa·s at 20°C could be developed, arginine was selected as a viscosity-reducing agent.

[0321] This test was carried out in three groups: the first group tested the crude pH and concentration; the second group tested the fine pH and concentration; and the third group tested the arginine concentration.

[0322] In the first series of studies, the effects of isatuximab concentration, pH, and temperature on viscosity were investigated. Seven formulations were prepared for this series of studies. Detailed compositions, along with the measured values, are shown in Table 6.

[0323] [Table 6]

[0324] A concentration of P188 at 0.4% w / v showed a positive stability effect compared to formulations without surfactant. A concentration of 2% sucrose allowed sufficient stability while maintaining osmolality close to isotonicity.

[0325] In the second set of studies, the effect of isatuximab concentration and pH on viscosity at 20°C was studied. A total of 32 formulations were prepared. Solutions were prepared at four target concentrations of antibody (126, 140, 147, and 154 g / L) and eight target pH values ​​(5.5, 5.7, 5.9, 6.2, 6.5, 6.7, 6.9, and 7.0). Detailed compositions, along with measured values, are shown in Table 7.

[0326] [Table 7]

[0327] In the third group of studies, the arginine concentration was varied between 90 and 150 mM. The detailed compositions of the five formulations are shown in Table 8, along with measured values ​​where available.

[0328] [Table 8]

[0329] For the first set of tests, the viscosity of all samples was determined with a Rheosense m-VROC viscometer at 5, 10, 15, 20, 25 and 30°C. For this purpose, the flow rate was selected at 50% of the maximum flow rate, and the time was 250-2500 s -1Newtonian behavior was assumed for all samples.

[0330] For the second group of tests, the viscosity of all samples was determined with a Rheosense m-VROC viscometer at 20 °C. For each sample, the flow rate was selected at 50% of the maximum flow rate determined by the instrument during the priming phase. This ranged from 250 to 2500 s -1 This led to a shear rate of 0.05. For all samples, Newtonian behavior was assumed.

[0331] For the third group of tests, the viscosity of all samples was determined with a Rheosense m-VROC viscometer at 20 °C. For each sample, the flow rate was selected at 50% of the maximum flow rate determined by the instrument during the priming phase. This was between 1200 and 1600 s -1 This led to a shear rate of 0.05. For all samples, Newtonian behavior was assumed.

[0332] Viscosity as a function of concentration At pH 5.5, the viscosity increased from 16 mPa·s at a concentration of 126 mg / mL to 54 mPa·s at 154 mg / mL. At pH 7.0, the viscosity increased from 5.8 mPa·s to 11 mPa·s over the same concentration range. For all conditions, the data were fitted using the Mooney equation:

number

[0333] As shown in Figure 8, the viscosity of the isatuximab solution increased with increasing mAb concentration.

[0334] Viscosity as a function of pH Figure 9 shows data obtained with m-VROC in Group 2, where viscosity was plotted as a function of pH for five concentrations of isatuximab (126, 140, 147, and 154 mg / mL). As shown in Figure 9, the viscosity of the isatuximab solution decreased with increasing pH.

[0335] At pH 6.2, viscosity was less than 25 mPa·s across the range of antibody concentrations tested. Viscosity was less than 25 mPa·s at all pH values ​​above 5.9. However, at the lower pH values ​​investigated (5.5 and 5.7), viscosity was greater than 25 mPa·s at mAb concentrations above 147 g / L.

[0336] Viscosity as a function of temperature In the first group, the viscosity of the investigated formulations was measured as a function of temperature from 5 to 30 °C. As shown in Figures 10 and 11, the viscosity decreased with increasing temperature. An Arrhenius fit was used to determine the activation energy of the formulations (Table 9). The values ​​were used in conjunction with a fit of the 20 °C data with the Mooney equation to calculate the theoretical values. As shown in Figure 10, the model matched the experimental data.

[0337] [Table 9]

[0338] The activation energy increased with increasing isatuximab concentration. The activation energy decreased at pH 5.9–6.2. To a lesser extent, the activation energy increased at pH 6.2–6.8. To move in solution, a mAb must escape from its neighbors and therefore requires a minimum energy, defined here as Ea. The probability that a mAb can acquire this energy is proportional to exp(–Ea / RT) according to Boltzmann's law; therefore, the viscosity, which is inversely proportional to the mobility of the mAb, follows the equation: viscosity = exp(+Ea / RT).

[0339] For the pH 6.2 formulation, at mAb concentrations of 143 and 126 mg / mL, the viscosity remained below 25 mPa·s in the temperature range of 5–30°C. At 154 mg / mL, the value recorded at 5°C was slightly higher than 25 mPa·s.

[0340] For the pH 5.9 formulation, the viscosity of 142 mg / mL isatuximab was approximately 25 mPa·s at 5°C.

[0341] Viscosity as a function of arginine concentration Preliminary studies showed that for a 150 mg / mL isatuximab formulation (without surfactant), viscosity decreased from 60 mPa·s to 16 mPa·s with 0–100 mM arginine and only to 12 mPa·s with 100–200 mM arginine.

[0342] In this study, arginine concentrations from 90 to 150 mM were tested at points of 90, 100, 115, 125, and 150 mM. Viscosity decreased with increasing arginine concentration (Figure 12). Values ​​decreased from 11 to 9 mPa·s, confirming the trend observed in the preliminary study.

[0343] Over the range of arginine concentrations tested (90–150 mM), the viscosity varied by less than 2 mPa·s. [Example]

[0344] stability This example describes a series of stability studies in which two sets of formulations were subjected to stability testing at 5° C. and accelerated stability testing at 40° C. / 75% relative humidity, as well as freeze-thaw and shake stress testing studies.

[0345] The formulations tested are summarized in Tables 10 and 11.

[0346] [Table 10]

[0347] [Table 11]

[0348] Freeze-thaw stress was performed using an Epsilon 1-6CC freeze-thaw unit (Martin Christ GmbH, Osterode, Germany). Samples were subjected to freeze-thaw stress.

[0349] One vial from each formulation (F4 and F10) was placed in a freeze dryer and cycled with the following parameters: speed, 0.1°C / min; freezing temperature, -30°C; thawing temperature, 25°C; number of cycles, 5; and hold-on temperature time, 60 min. After five cycles, the samples were inspected for visible particles and homogenized.

[0350] One vial of each formulation was mounted onto a horizontal shaking platform (IKA, KS4000 IC) and stressed for 21 days at 25°C and 300 rpm. T-mech samples were analyzed along with the T-1 month time point.

[0351] The osmolality of the samples was measured by the freezing point depression method using a Gonotec Osmomat 3000 (Gonotec, Berlin, Germany). Prior to running the instrument, a three-point calibration was performed, which included Milli-Q water and two osmolality standards of 300 and 400 mOsmol / kg.

[0352] Protein concentrations were determined by UV spectroscopy performed in a 96-well plate (Corning Incorporation, NY, USA) on a Tecan Safire2 plate reader (Tecan Austria GmbH, Grodig, Austria). Samples were gravimetrically diluted from a 150 mg / mL solution to a protein concentration of 1 mg / mL. The dilution factor was calculated from the balance printout. For each data point, three wells (n=3) filled with 200 μL of solution were measured to minimize measurement error. The temperature of the measurement cell was set to 25°C. The dilution buffer was measured as a blank spectrum. After measurement, the absorbance values ​​obtained at 280 nm were corrected for path length and subtracted by the corresponding blank. The calculated molar extinction coefficient (ε) at 280 nm was calculated as モル )224,320M -1 cm -1 was used to calculate the extinction coefficient (ε). To determine the protein concentration based on the absorbance value at 280 nm, 1.548 mL mg -1 cm -1 The calculated extinction coefficient (ε) was used.

[0353] Vials were inspected for the presence of visible particles according to the European Pharmacopoeia (8th edition; Monograph 2.9.20) at approximately 3750 lux for 5 seconds against a white background and for 5 seconds against a black background under weak manual radial agitation. The inspections were performed independently by two trained inspectors.

[0354] A numerical score based on the "Deutscher Arzneimittel-Codex" (DAC 2006) was used to classify the observed visible particles (0, no visible particles within 5 seconds; 1, few visible particles within 5 seconds; 2, medium number of visible particles within 5 seconds; 10, many particles directly visible). Fibrous structures and particles that are probably non-specific to the product are not accounted for by the numerical score.

[0355] High-Performance Size Exclusion Chromatography (HP-SEC) Prior to sample analysis, the performance of the HP-SEC column was tested with BioRad gel filtration standards (containing thyroglobulin, gamma globulin, ovalbumin, myoglobin, and vitamin B12). The lyophilized material was solubilized in 500 μL of Milli-Q water, followed by a 10-fold dilution with mobile phase (final protein concentration of 3.6 mg / mL). System suitability tests were performed at the beginning of each sequence by injecting the gel filtration standard (BioRad) and calculating the USP resolution between the gamma globulin and ovalbumin peaks. A 50 mg / mL stock solution sample was diluted 5-fold in a 1.5 mL polypropylene tube (Eppendorf) by mixing 40 μL of the stock solution with 160 μL of solution A (mobile phase without acetonitrile), resulting in a protein concentration of 10.0 mg / mL.

[0356] To avoid blocking of the HP-SEC column by larger insoluble particles that could potentially form during accelerated stability / stress testing, after dilution the samples were centrifuged at 18,000 rcf for 5 minutes and the supernatant transferred to HPLC vials. The samples were vortexed and stored at 5°C in the autosampler until analysis.

[0357] The following parameters were used for the HP-SEC analysis: Equipment: Ultimate 3000 (Dionex) Column: amorphous silica column ProSEC 300S; 300 mm Security guard: Guard column 50mm x 7.5mm Flow rate: 0.3mL / min Mobile phase: 90% 100 mM phosphoric acid, pH 7.2, 300 mM NaClO4 + 10% ACN Detection: UV at 280nm and 214nm Column oven: 25℃ Sample cooling: 5 to 8°C Injection volume: 10 μL for 5.0 mg / mL samples, 10 μL for blanks and SEC standards Analysis time: 90 minutes

[0358] Capillary isoelectric focusing (cIEF) Imaging capillary isoelectric focusing (cIEF) was performed on an iCE280 instrument connected to a PrinCE microinjector (Convergent Bioscience, Toronto, Canada). Instead of eluting a focal molecular species past a fixed detection time point, as is done in conventional cIEF, in imaging cIEF, molecules are detected throughout the entire IEF capillary. To do so, ultraviolet light at a wavelength of 280 nm was focused onto a UV-transparent capillary, and images were captured at regular intervals with the aid of a charge-coupled device (CCD) camera.

[0359] Prior to analysis, the fused silica-coated (FC) cartridge was installed according to the instrument instructions. The anode reservoir was filled with 0.08 M phosphoric acid (in 0.1% methylcellulose, electrolyte kit, ProteinSimple), and the cathode reservoir was filled with 0.1 M sodium hydroxide (in 0.1% methylcellulose, electrolyte kit, ProteinSimple). System performance was verified by measuring a hemoglobin standard (iCE280 System Suitability Kit, ProteinSimple). Isoelectric focusing of the hemoglobin standard solution was performed according to the manufacturer (prefocusing: 1 min at 1500 V; focusing: 4.5 min at 3000 V).

[0360] Prepare a Master Sample for 20 samples by mixing 2360 μL of Milli-Q water, 1400 μL of 1% methylcellulose, 160 μL of Pharmalyte pH 3–10, 20 μL of pI marker 7.05, and 20 μL of pI marker 9.50. The master mix was homogenized by vortexing and briefly centrifuged at 5,000 rcf, and the mixture was filtered through a 0.45 μm syringe PVDF filter unit (Millex-GV, Millipore).

[0361] Samples were pre-diluted to a protein concentration of 20.0 mg / mL by mixing 20 μL of the stock solution (C=50 mg / mL) with 30 μL of the corresponding formulation buffer. Final samples for cIEF analysis were prepared by mixing 2 μL of the 20.0 mg / mL pre-diluted sample with 198 μL of Master Mix to obtain a total volume of 200 μL and a protein concentration of 0.2 mg / mL.

[0362] Isoelectric focusing of DP was performed by prefocusing at 1500 V for 1 min followed by focusing at 3000 V for 8 min. UV absorption images were captured using the software ChromPerfect (5th edition). The analysis was performed using the .5.6 version.

[0363] Visual inspection results Visual inspection was performed by two independent operators. Sample solutions after freeze-thawing were marked as inhomogeneous (S-Schlieren*, phase separation). No major changes in visible particle content and turbidity were observed for all test formulations during the stability study.

[0364] The results of the osmolality measurements are shown in Table 12. At TO, osmolality varied from 275 to 418 mOsmol / kg. Formulation F10 showed higher osmolality values ​​than formulation F4. Storage at 40°C / 75% relative humidity did not affect osmolality in the test samples.

[0365] [Table 12]

[0366] [Table 13]

[0367] The results of the viscosity measurement at TO showed that the F10 formulation exhibited a slightly higher viscosity than the F4 formulation. The highest viscosity was observed for formulation F4-8 (1000 s at 20 °C). -1 30.03cP) and F10-8 (1000s at 20°C)-1 In both sets of formulations (F4 and F10), run 8 contained the highest protein concentration (154 mg / mL), the lowest pH (5.9), and the highest sucrose concentration (24 mg / mL).

[0368] The results of protein concentration determination by ultraviolet spectroscopy are provided in Tables 14 and 15. At TO, the protein concentration ranged from 124 to 156 mg / mL. The protein concentration in the test formulations remained stable during storage at 40°C / 75% relative humidity.

[0369] [Table 14]

[0370] [Table 15]

[0371] Sodium dodecyl sulfate gel electrophoresis (SDS-PAGE) was used to characterize the molecular weight and relative amounts of protein species. The relative amounts of separated species were calculated from SDS-PAGE gels by measuring the optical density of the detected protein bands. The relative amounts and molecular weights of all detected protein species under native and reducing conditions are found in Tables 16 and 17, respectively.

[0372] [Table 16]

[0373] [Table 17]

[0374] High-performance size-exclusion chromatography to investigate monomers, aggregates, and fragments HP-SEC was performed and four aggregate species (HMW1, HMW2, HMW3, and HMW4) and two fragment species (LMW1 and LMW2) were assigned.

[0375] At TO, the relative monomer content in formulations F4 and F10 ranged from 98.5 to 99.0%. Storage at 40°C / 75% relative humidity led to a decrease in monomer content in all test samples. After one month of storage at 40°C / 75% relative humidity, the monomer content in the test formulations ranged from 96.2 to 97.2%; the lowest monomer content at this time point was found in formulation F4-2. Repeated freeze-thaw cycles had little effect on the relative monomer content, while mechanical stress led to a small decrease (97.7 to 98.2%). See Figures 13 and 14.

[0376] For almost all samples and time points, the relative content of HMW1 was below the limit of quantitation of 0.15%, and similarly for almost all samples and time points, the relative content of HMW2 was below the limit of quantitation of 0.15%.

[0377] The relative HMW3 content ranged from 0.7 to 0.9% at TO. After 1 month of storage at 40°C / 75% relative humidity, the HMW3 content increased to 1.1 to 1.5%. The largest increases in HMW3 content were observed in samples F4-14, F10-5, F10-12, and F10-14, whereas the smallest increases were observed in samples F4-1 and F4-7. Repeated freeze-thaw cycles did not result in an increase in the HMW3 relative content, but mechanical stress led to a slight increase in HMW3 to 0.9 to 1.2%.

[0378] For almost all samples and time points, the relative content of HMW4 was below the limit of quantification of 0.15%.

[0379] The sum of all aggregates (HMWS) was calculated for peaks ≥ 0.15% relative area. At TO, the total aggregate (HMWS) content ranged from 0.6 to 1.2%. Storage at 40°C / 75% relative humidity led to a significant increase in HMWS content in all test formulations. After 1 month of storage at 40°C / 75% relative humidity, the relative HMWS content ranged from 1.1 to 1.9%. The smallest increase in HMWS species was observed in formulation F4-7, while the largest increases were observed in formulations F4-2 and F10-14. No substantial change in HMWS content was observed after freeze-thawing, and only a small increase was detected after exposure to mechanical stress, leading to HMWS contents of 0.9 to 1.3%. See Figures 15 and 16.

[0380] At TO, the content of fragment LMW1 was approximately 0.3. Storage at 40°C / 75% relative humidity led to an increase in LMW1 content in all test samples. The relative LMW1 content was highest after 1 month of storage at 40°C / 75% relative humidity and ranged from 1.1 to 1.5%. Freezing and thawing did not affect the LMW1 content. Mechanical stress led to a small increase in the LMW1 content.

[0381] At T0, for all test formulations, the relative content of fragment LMW2 was below the limit of quantification (≥0.15%). Storage at 40°C / 75% relative humidity for 2 weeks and 1 month led to a small increase in LMW2 content, with the highest value observed at T-1m_40°C (0.3-0.5%).

[0382] The sum of all LMWS contents was calculated with a peak ≥ 0.15%. At T0, the total LMWS content was approximately 0.3% for all test formulations. Freeze-thawing did not affect LMWS content, but mechanical stress led to a small increase in LMWS content (0.7-0.9%). Storage at 40°C / 75% relative humidity led to a more significant increase, especially at later time points (Figures 17 and 18). The largest increase was observed at T-1m_40°C, where the total LMWS content The amount varied in the range of 1.4-1.8%.

[0383] At T0, the relative content of the main peak, as determined by capillary isoelectric focusing (cIEF), ranged from 70.3 to 74.0%. After storage at 40°C / 75% relative humidity for 1 week, 2 weeks, and 1 month, a gradual decrease in the relative content of the main peak was detected. The greatest decrease was observed in formulation F10-8 (55.4% at T-1m_40°C). The pI of the main peak remained stable at approximately 8.2 for all formulations at all stability time points.

[0384] At T0, the relative content of acidic species varied between approximately 17.9 and 20.9%. Storage at 40°C / 75% relative humidity led to a significant increase in acidic species content. The largest increase was observed at T-1m_40°C, with values ​​ranging from 30.4 to 36.9%. The largest increase was observed in formulation F10-8, while formulation F4-7 showed the smallest increase. Graphical representations of the data are shown in Figures 19 and 20.

[0385] At T0, a basic peak content of 8.1-9.5% was observed. During the stability test, the basic peak content remained relatively stable for all formulations at most time points. At T-1m_40°C, the basic peak content varied from 7.1-9.1%.

[0386] After storage at 40°C / 75% relative humidity, capillary isoelectric focusing (cIEF) data showed an increase in acidic species content at the expense of the main peak content for all formulations. The F4 formulation exhibited a smaller decrease in the main peak relative content after 1 month of storage at 40°C / 75% relative humidity, demonstrating better chemical stability than the F10 formulation, as shown in Figure 21. After 1 month of storage at 40°C / 75% relative humidity, the F4 formulations exhibited a monomer peak loss ranging from 9.5 to 13.9%, while the F10 formulation exhibited a monomer peak loss ranging from 14.7 to 17.3%. [Example]

[0387] stability Results are provided for a stability study performed on 140 mg / mL isatuximab formulated in 110 mM arginine-HCl, 9 mM histidine, 2% sucrose, 0.4% poloxamer 188, pH 6.2.

[0388] result Stability at -20°C ± 5°C As shown in Table 18, after one month of storage at -20°C ± 5°C, all quality attributes tested, visible particles, color, degree of opalescence, purity by SEC and cGE, protein concentration by UV, potency by ADCC and CDC bioassays, particulate matter by darkness, and pH were stable.

[0389] Charge heterogeneity by icIEF did not show any significant changes compared to the starting material.

[0390] Stability at +5°C±3°C (long-term storage conditions) As shown in Table 19 and Table 20, after 1, 3, 6, 9 and 12 months of storage at +5°C ± 3°C, all quality attributes tested, visible particles, purity by SEC and cGE, protein concentration by UV, potency by CDC bioassay, particulate matter by darkness and pH, were stable for at least 12 months.

[0391] The color, degree of opalescence and charge heterogeneity by icIEF did not show any significant changes compared to the starting material for at least 12 months.

[0392] Stability at +25°C±2°C / 60%±5%RH (accelerated storage conditions) As shown in Table 21, after 6 months of storage at +25°C ± 2°C, all quality attributes tested (visible particles, purity by SEC, protein concentration by UV, potency by ADCC and CDC bioassays, particulate matter by darkness, and pH) were stable for at least 6 months.

[0393] After 6 months at +25°C ± 2°C, no changes were observed in both color and degree of opalescence.

[0394] After 6 months at +25°C ± 2°C, the following changes in the charge heterogeneity profile by icIEF were observed: a 9% decrease in the main isoform content together with a 9% increase in the acidic isoform was observed.

[0395] A slight decrease in the main peak purity by cGE (-2%) was observed to correlate with an increase in the total low molecular weight species (+2%).

[0396] A slight decrease in monomer purity by SEC (-1.7%) was observed.

[0397] All other parameters did not show any significant changes compared to the starting material.

[0398] Stability at +40°C±2°C / 75%±5%RH (stress conditions) As shown in Table 22, the following changes were observed after one month under stress conditions: The following changes in the charge heterogeneity profile by icIEF were observed: a 14% decrease in the main isoform content, which correlated primarily with a 15% increase in the acidic form.

[0399] A slight decrease in the main peak purity by cGE (-2%) was observed to correlate with an increase in the total low molecular weight species (+2%).

[0400] The percentage of aggregates by SEC remained consistent.

[0401] conclusion The results of this stability study indicate that 140 mg / mL isatuximab formulated in 110 mM arginine-HCl, 9 mM histidine, 2% sucrose, 0.4% poloxamer 188, pH 6.2 is stable at -20°C for at least 1 month and at +5°C ± 3°C for at least 12 months.

[0402] [Table 18]

[0403] [Table 19]

[0404] [Table 20]

[0405] [Table 21]

[0406] [Table 22] [Example]

[0407] In Vivo Studies in Minipigs This example describes a study in which minipigs were treated with a formulation of isatuximab administered by subcutaneous injection. Minipigs were chosen as the test model for this study of subcutaneous local tolerance because of their well-tolerated suitability and because they are commonly used for nonclinical evaluation of intended human administration routes.

[0408] Four formulations (F1, F2, F4, and F10) and saline (0.9% sodium chloride) as a negative control were tested in eight minipigs (1-year-old females, 20-25 kg), with four animals per test group. Each animal received two injections of the formulation and the negative control, with a 3-week recovery period between injections. Administration was performed in the flank via a catheter with a butterfly 27G needle using a syringe pump (Harvard Apparatus Model "22") equipped with a backpressure captor (RSB5 Subminiature Load Cell 50 lb / 200 N) for tissue backpressure monitoring.

[0409] During the injection, observe the animals focusing on general behavior, vocalizations, and visual parameters for pain assessment. Injection syringe pressure and evidence of leakage were also monitored. Injection sites were observed for skin changes, and plasma analysis was performed for substance P and cortisol. Skin biopsies were collected in 10% NBF on day 5.

[0410] Pain symptoms were recorded during and after injection for 5 days. Blood was collected along the injection and for the following 90 minutes and dosed with pain markers. Skin samples taken at the time of injection were then subjected to histopathological examination. With regard to histopathology, all formulations were well tolerated.

[0411] All solutions of isatuximab were prepared from the same pre-formulated batch of isatuximab solution, which was formulated at 30 mg / mL, pH 6, with 20 mM histidine and 5% sucrose.

[0412] All formulations tested for the research minipig study are listed in Table 23.

[0413] [Table 23]

[0414] For the preparation of each formulation, the buffer (without sucrose and surfactant) was exchanged by ultrafiltration to concentrate the antibody above the target formulation. After an initial adjustment of the concentration, a concentrated excipient solution (concentrated sucrose and surfactant dissolved in ultrafiltration buffer) was added to obtain the final formulation.

[0415] The ultrafiltration buffers tested are listed in Table 24.

[0416] [Table 24]

[0417] The recorded values ​​for the final formulation are shown in Table 25.

[0418] [Table 25]

[0419] A total of eight one-year-old female minipigs were used in the study. Each formulation was tested on four animals, each receiving 18 mL of infusion over 30 minutes (0.6 mL / min). The study lasted five days, during which pain markers and symptoms were recorded. At the end of the five days, 8 mm discs of skin were taken for histopathology (biopsy).

[0420] Formulations F2 and F4 were first tested in minipigs. After a lag time of 21 days, formulations F1 and F10 were then tested in the same eight minipigs. Finally, after a second lag time, saline was tested in the same eight minipigs.

[0421] For each formulation / saline solution, testing was performed as follows: Injection of the solution into 4 minipigs (8 in saline solution) During the injection: Live recording of the following parameters: Telemetry measurement (ECG) Syringe back pressure (to calculate tissue back pressure) Pain markers (cortisol, substance P, blood medication - regular blood sampling required) Pain symptoms (scratching, rubbing, noise, redness) and size of edema from injection. ·After injection: Pain markers 90 minutes after the end of the infusion Pain symptoms (scratching, rubbing, redness) for 5 days (visual inspection, arbitrary units) Histopathology of sample tissue (skin biopsy after 5 days)

[0422] The main results regarding pain are summarized in Table 26.

[0423] [Table 26]

[0424] In Table 26, only plasma cortisol was reported as a pain marker. The levels of substance P measured for all formulations did not change over time. An increase in plasma cortisol was recorded after injection. The maximum mean value calculated across the four minipigs for each formulation is reported in Table 26. The maximum mean value was obtained with F10. For all formulations, there was variability in plasma cortisol levels among the four test animals.

[0425] The number of pain symptoms for F10 was also the highest of the four formulations and had the highest plasma cortisol levels (Figure 22). F2 had the second highest number of pain symptoms and the highest substance P levels (Figure 23). The finding that both F1 and F2 had poor antibody stability was not shown in Table 26.

[0426] No leakage and fairly constant syringe back pressure was observed during injection of the four formulations.

[0427] The main histopathology results are summarized in Table 27.

[0428] [Table 27]

[0429] The dermis and subcutaneous tissue showed no significant differences between the formulations. Formulation F2 was significantly different from Formulation F1. (25%), showed a higher incidence of skeletal muscle changes (100%) compared to F4 (25%), F10 (0%) or saline (25%). [Example]

[0430] Second in vivo study in minipigs The purpose of the study described in this example was to investigate the local tolerance and plasma pharmacokinetics of isatuximab after a single administration by subcutaneous infusion in minipigs using three different flow rates. Additionally, one group of animals was dosed intravenously once to assess the bioavailability of isatuximab after different subcutaneous infusions.

[0431] Test samples used were isatuximab 500 mg / 25 mL (20 mg / mL) for IV infusion and isatuximab 140 mg / mL for SC infusion (Formulation F4 in Example 5). Saline for injection (0.9% NaCl) was used as the negative control for groups 2, 3, and 4.

[0432] The study was performed on 20 female Göttingen SPF (specific pathogen-free) minipigs from Ellegaard Göttingen Minipigs A / S, DK-4261 Dalmose, Denmark. Animals were ordered at a weight of 20–25 kg upon arrival. A pre-treatment period of 15 days (including a 5-day acclimatization period) was allowed, during which the animals were observed daily to exclude any animals in poor condition. All observations were recorded.

[0433] Animals were randomized into four treatment groups as shown in Table 28:

[0434] [Table 28]

[0435] For intravenous infusion, ear vein catheters were implanted in six animals (5 + 1 spare) 9 days before the start of treatment. Five of these animals were assigned to Group 1 (IV infusion group); the last animal was included in one of the subcutaneous groups.

[0436] All administrations were performed using a Baxter Colleague CXE volumetric infusion pump, which is capable of infusing from semi-rigid containers, rigid containers, flexible IV bags, and vented syringes. Test and negative control articles were placed in sterile glass infusion bottles during administration.

[0437] The first day of treatment was designated as day 1. On day -1, Group 1, a single dose of isatuximab (1800 mg / animal) was given by 30-minute infusion at a flow rate of 3 mL / min via an implanted ear vein catheter. On day -1, groups 2, 3, and 4 received a single dose of isatuximab (1806 mg / animal) at flow rates of 0.5, 1, and 2 mL / min, respectively, via a subcutaneous catheter with a butterfly needle placed in the left lower quadrant just before the knee. On day -8, groups 2, 3, and 4 received a single dose of saline (negative control) at flow rates of 0.5, 1, and 2 mL / min, respectively, via a subcutaneous catheter with a butterfly needle placed in the right lower quadrant just anterior to the knee. The dose was 90 mL in Group 1 and 12.9 mL in Groups 2-4. During the infusion, the subcutaneous injection sites of Groups 2-4 were observed for leakage. - The subcutaneous injection site was marked along the edge of the local swelling that occurred during the injection procedure and remarked as necessary. -The needle size for SC injection was 23G.

[0438] All signs of ill health and any behavioral changes were recorded daily. Any deviations from normal were noted. During dosing, animals were observed for general behavior and any vocalizations, with emphasis on any signs of stress, discomfort, or pain.

[0439] For Groups 2-4, the subcutaneous injection sites were observed daily from the day of administration for bleeding, erythema, swelling (blebbing, indicated by size), and firmness, but not excluding other signs. On the day of administration, the injection sites were observed before and at the end of the infusion, and then 15 minutes (± 2 minutes), 30 minutes (± 2 minutes), 1 hour (± 3 minutes), 2 hours (± 6 minutes), and 4 hours (± 12 minutes) after the end of administration. Thereafter, injection sites were observed daily until day 17, as no local reactions were observed from day 10. Parameters were assessed by the following grading system: 0 - absent; 1 - minimal; 2 - slight; 3 - moderate; and 4 - marked.

[0440] Blood samples for pharmacokinetics Blood samples were taken from all animals starting on day 1. Blood collection was performed at the following time points: pre-treatment, within 2 minutes of the end of the infusion, 1 (±3 minutes), 4 (±12 minutes), 24 (±1 hour 12 minutes), 48 (±2 hours 24 minutes), 72, 96, 168, 192, 264, 336, 504, and 672 hours after the end of the infusion (72 hours ±3 hours tolerance).

[0441] Pharmacokinetic (Pk) analysis was performed using the software Phoenix WinNonlin version 6.3 by Pharsight Corporation, Mountain View, CA, USA. Where appropriate, non-compartmental analysis was performed using WinNonlin plasma models (intravenous infusion and extravascular administration models).

[0442] Plasma concentration-time data from each individual animal were used for pharmacokinetic calculations. In addition to parameter estimates for individual animals, descriptive statistics (e.g., mean, standard deviation, and coefficient of variation) were reported where appropriate. All parameters for each animal were generated from the individual test article concentrations in plasma after treatment on Day 1. For the determination of individual pharmacokinetic parameters, concentrations below the limit of quantitation were treated as zero. For the determination of mean concentrations, samples below the limit of quantitation were treated as zero.

[0443] Parameters were estimated using nominal dose levels. Parameters were estimated using nominal sampling times, as no time deviations from nominal of more than 15% were demonstrated. Pre-dose concentrations on Day 1 were set to zero.

[0444] Descriptive statistics (means, standard deviations, where applicable) and pharmacokinetic parameters were reported to three significant figures. Coefficients of variation were reported with no decimal places.

[0445] The following PK parameters were estimated for isatuximab: C max - maximum observed concentration, ·t max - time of maximum concentration, C last - last measurable concentration, ·t last - time of last measurable concentration, AUC 0~24時間 The area under the plasma concentration-time curve from 0 to 24 hours was calculated by non-compartmental analysis using the linear trapezoidal rule. AUC 0~72時間 The area under the plasma concentration-time curve from 0 to 72 hours was calculated by non-compartmental analysis using the linear trapezoidal rule. AUC 0~168時間 The area under the plasma concentration-time curve from -0 to 168 hours was calculated by non-compartmental analysis using the linear trapezoidal rule. AUC 0~t The area under the plasma concentration-time curve calculated from 0 to t, where t is the time of the last measurable concentration, was calculated by non-compartmental analysis using the linear trapezoidal rule. The bioavailability of SC injection compared with IV injection was evaluated by AUC 0~24時間 , AUC 0~72時間 , AUC 0~168時間 AUC 0~t was evaluated using.

[0446] Further parameters (i.e., t 1 / 2z , V z ., CL) was calculated for the IV route.

[0447] Collection of skin biopsies from SC injection sites (Groups 2–4) On both days 8 and 29, three skin biopsies approximately 7-10 mm deep were taken from the left subcutaneous injection site (test article site) using a 6 mm biopsy punch for histopathology. The day 8 collection was performed after administration of saline (negative control) so that animals could be monitored during the injection.

[0448] Biopsies collected on day 8 (biopsies 1-3) were taken from the cranial half of the injected area. Biopsy 1 was collected from the dorsal region, biopsy 2 from the central region, and biopsy 3 from the ventral region. Biopsies collected on day 29 were similarly collected from the caudal half of the injected area. Additionally, an untreated control (biopsy 4) from outside the injected area was collected from the same region in all animals on both days.

[0449] On both days 15 and 36, skin biopsies were similarly collected from the right subcutaneous injection site (saline site) of all animals in groups 2-4.

[0450] Biopsies collected on day 15 (biopsies 5-7) were taken from the cranial half of the injected area. Biopsy 5 was collected from the dorsal region, biopsy 6 from the central region, and biopsy 7 from the ventral region. Biopsies collected on day 36 were similarly collected from the caudal half of the injected area. Additionally, an untreated control (biopsy 8) from outside the injected area was collected from the same region in all animals on both days.

[0451] Each biopsy from each animal was placed in a separate container and fixed in phosphate-buffered neutral 4% formaldehyde.

[0452] result No test article-related clinical signs were observed in any of the animals.

[0453] In three animals (number 4, group 1, and numbers 6 and 7, group 2), the skin appeared warm to the touch on days 1 and 2. However, because the number of affected animals was low, this was considered an incidental finding.

[0454] In all groups, local reactions at the injection site were assessed on the day of injection. No scores were obtained on other days.

[0455] On Day 1 (isatuximab), in Group 2 (0.5 mL / min), slight erythema was noted in all animals, primarily at the infusion site, and minimal to moderate swelling (blebbing) was observed in three animals within the first two hours after the end of the infusion. Additionally, three animals had minimal bleeding within the first 15 minutes after the end of the infusion.

[0456] In Group 2 on Day 8 (0.5 mL / min saline), minimal erythema was noted in all animals, primarily at the infusion site, and minimal to moderate swelling (blebbing) was observed in two animals within the first two hours after the end of the infusion. Additionally, minimal bleeding was observed in three animals after the end of the infusion.

[0457] On Day 1 (isatuximab), Group 3 (1 mL / min) showed minimal to slight erythema at the infusion site within the first 4 hours after the end of the infusion in 4 animals, and minimal to significant swelling (blebbing) was seen in 3 animals, decreasing in size by 4 hours after the end of the infusion. Minimal bleeding, and slight bleeding in 1 animal, was observed in 3 animals within the first 30 minutes after the end of the infusion.

[0458] On Day 8 (1 mL / min saline) in Group 3, minimal to slight erythema was observed at the infusion site in all animals within the first 4 hours after the end of the infusion, and one animal had minimal to moderate swelling (blebbing) after administration that decreased in size by 2 hours after the end of the infusion. Minimal bleeding, and slight bleeding in one animal, was observed in all animals within the first 15 minutes after the end of the infusion.

[0459] On Day 1 (Isatuximab), Group 4 (2 mL / min) showed minimal to slight erythema (moderate in one animal) at the infusion site within the first 2 hours of the end of the infusion in four animals, with up to significant swelling (blebbing) seen in one animal post-dose, decreasing in size by 4 hours after the end of the infusion. Two animals showed minimal bleeding at the end of the infusion.

[0460] On Day 8 (2 mL / min saline) in Group 4, minimal (slight in 2 cases) erythema was observed in all animals within the first 30 minutes of the end of the infusion, primarily at the infusion site, and at most moderate swelling (blebbing) was seen in 2 animals post-dose, decreasing in size by 1 hour after the end of the infusion. Minimal bleeding was observed in 2 animals at the end of the infusion.

[0461] No signs of leakage were observed during the infusion.

[0462] [Table 29]

[0463] Pharmacokinetics The individual plasma concentrations of isatuximab over time are shown in Figures 24-27.

[0464] Non-compartmental pharmacokinetic analysis of plasma level data was performed using Phoenix WinNonlin version 6.3 pharmacokinetic software.

[0465] [Table 30]

[0466] Isatuximab was quantifiable in all plasma samples in the study collected after the end of infusion, except for a single sample taken at the end of SC infusion in Group 4 females and 1 hour after SC infusion in Group 3 females, which was below the lower limit of quantification (LLOQ). Concentrations in all predose samples were below the LLOQ. For the SC and IV routes, profiles were consistent with extravascular and intravenous administration, respectively. Overall, the variability observed in PK parameters was low with IV infusion and low to moderate with SC infusion.

[0467] After a single IV infusion of 1800 mg isatuximab per animal into minipigs over 30 minutes, maximum plasma levels were observed in all animals at the end of the infusion period. After a single SC infusion of 1806 mg isatuximab per animal into minipigs at flow rates of 0.5, 1, or 2 mL / min, median maximum plasma levels were observed at 96, 192, and 168 hours from the end of the infusion, respectively. However, individual T max The values ​​ranged from 48 to 264 hours and showed no relationship with SC infusion rate.

[0468] Following a single IV infusion of 1800 mg isatuximab per animal over 30 minutes into minipigs, the mean AUC (AUC last ) was 364,000 h*μg / mL. After a single SC infusion of 1806 mg isatuximab per animal into minipigs at flow rates of 0.5, 1, or 2 mL / min, the mean AUC (AUC last ) were 326,000, 565,000, and 369,000 h*μg / mL, respectively. AUC last Values ​​were generally similar for each SC group, suggesting that infusion rate did not affect exposure.

[0469] Under the conditions of this study, when given by SC infusion to minipigs at flow rates of 0.5, 1, or 2 mL / min, the absolute SC bioavailability of isatuximab was broadly similar at the three flow rates tested at each of the AUC intervals considered. The AUC (AUC last ), the bioavailability increased with increasing AUC interval, being 0.89, 1.55, and 1.05 at flow rates of 0.5, 1, and 2 mL / min. F values ​​of 1.01 and 1.02 were reached, respectively. In the group infused at 1 mL / min, the estimated higher F values ​​were related to the higher variability noted in this group; contributed by 2 / 5 animals that showed a clearly higher exposure.

[0470] Overall, it was concluded that the absolute SC bioavailability of isatuximab in minipigs was at least 89% when given at a dose of 1806 mg (140 mg / mL solution) per animal by SC infusion at a flow rate of 0.5–2 mL / min.

[0471] Microscopic examination of a skin biopsy Microscopic examination of injection site skin biopsies revealed no treatment-related changes in any of the subcutaneous groups. Microscopic findings consisted primarily of minimal focal infiltration of mononuclear cells and minimal focal epidermal crusts, with no differences in incidence or morphology of findings between subcutaneous treatment groups. Similarly, findings were considered similar when saline (negative control)-treated injection sites were compared with test sample-treated injection sites within treatment groups (at the same infusion rate). All microscopic findings were considered to be well-known incidental background changes in the skin of Göttingen minipigs.

[0472] Consideration Isatuximab was clinically and histopathologically very well tolerated when given to female Göttingen minipigs by intravenous infusion (as a 20 mg / mL solution) at a dose of 1800 mg per animal and a flow rate of 3 mL / min, or as a subcutaneous infusion (as a 140 mg / mL solution in Formulation F4 of Example 5) at a dose of 1806 mg per animal and a flow rate of 0.5, 1, or 2 mL / min.

[0473] Local reactions at the subcutaneous injection site in all groups were only observed at the site on the day of injection.

[0474] The incidence and severity scores of erythema and bleeding were comparable among the three infusion rates tested (0.5, 1, and 2 mL / min) and were similar for isatuximab and the saline negative control. However, swelling (blebbing) at the injection site was more pronounced with isatuximab infused at 1 mL / min and 2 mL / min compared with 0.5 mL / min. This was likely a physical phenomenon related to the large volume injected into a single site, and severity was inversely proportional to infusion time at a constant rate of fluid removal from the injection site. Furthermore, swelling was more pronounced after isatuximab infusion than after saline infusion. Swelling after saline infusion was comparable at all three infusion rates.

[0475] Microscopic examination of injection site skin biopsies showed no treatment-related changes in any of the subcutaneous groups.

[0476] After a single IV infusion of 1800 mg isatuximab per animal into minipigs over 30 minutes, maximum plasma levels were observed in all animals at the end of the infusion period. After a single SC infusion of 1806 mg isatuximab per animal into minipigs at flow rates of 0.5, 1, or 2 mL / min, median maximum plasma levels were observed at 96, 192, and 168 hours from the end of the infusion, respectively. However, individual T max The values ​​ranged from 48 to 264 hours and showed no relationship with SC infusion rate.

[0477] Following a single IV infusion of 1800 mg isatuximab per animal over 30 minutes into minipigs, the mean AUC (AUC last ) was 364,000 h*μg / mL. After a single SC infusion of 1806 mg isatuximab per animal into minipigs at flow rates of 0.5, 1, or 2 mL / min, the mean AUC (AUC last ) is that The AUC values ​​were 326,000, 565,000, and 369,000 h*μg / mL, respectively. lastValues ​​were generally similar for each SC group, suggesting that infusion rate did not affect exposure.

[0478] Under the conditions of this study, when given by SC infusion to minipigs at flow rates of 0.5, 1, or 2 mL / min, the absolute SC bioavailability of isatuximab formulated in F4 of Example 4 was broadly similar at the three flow rates tested in each of the AUC intervals considered. The AUC (AUC last When calculated using the α-value (AUC) function, bioavailability increased with increasing AUC interval, reaching F values ​​of 0.89, 1.55, and 1.01 at flow rates of 0.5, 1, and 2 mL / min, respectively. In the group infused at 1 mL / min, the estimated higher F values ​​were related to the higher variability noted in this group, contributed by 2 / 5 animals that showed a clearly higher exposure. [Example]

[0479] A Phase 1b Study of Subcutaneous Isatuximab in Humans This example describes a multicenter, open-label, phase 1b study to investigate the pharmacokinetics, safety, and efficacy of subcutaneous and intravenous isatuximab in combination with pomalidomide and dexamethasone in patients with relapsed / refractory multiple myeloma (RRMM).

[0480] This study was designed to investigate SC administration of isatuximab for the first time. Furthermore, the SC formulation was not the same as the formulation used for IV administration. SC isatuximab in combination with pomalidomide and dexamethasone was administered to a similar patient population as in the previous study noted above. To allow for investigation of the safety and PK of SC isatuximab, this study also included a cohort with IV isatuximab administration.

[0481] The primary endpoints of this study were (i) to evaluate the safety and tolerability (including local injection site tolerance) of isatuximab administered subcutaneously (SC) using an infusion pump versus intravenously (IV) isatuximab; and (ii) to evaluate the pharmacokinetics of isatuximab when administered SC and IV in combination with pomalidomide and dexamethasone. Secondary endpoints of this study included (i) estimating the absolute bioavailability of isatuximab SC and IV; (ii) measuring the CD38 receptor occupancy (RO) of isatuximab on plasma cells from bone marrow aspirates after SC versus IV administration; and (iii) evaluating the efficacy of isatuximab SC / IV administration.

[0482] The study included five cohorts. Patients were randomized to cohort 1a (1000 mg SC dose) or 1b (IV) (2:1 randomization ratio). After evaluation of safety, PK, and ROI data for SC isatuximab in cohort 1a, additional patients were randomized to cohorts 2a or 2b (2:1 randomization ratio), with a higher dose of SC isatuximab in cohort 2a (1400 mg dose) and the same IV dose in cohort 2b. Once all patients in cohorts 2a and 2b completed cycle 1 of treatment, a final review of safety, PK, and ROI data following administration of SC and IV isatuximab was performed. After confirmation of the recommended phase 2 dose (RP2D) level, an additional 22 patients were recruited to cohort 2c and received SC isatuximab at this dose level. Table 31 describes treatment by cohort in further detail.

[0483] [Table 31]

[0484] Isatuximab was administered in combination with pomalidomide and dexamethasone once weekly for 4 weeks (Cycle 1) and on days 1 and 15 of each subsequent cycle. Each cycle was 28 days in duration. All study participants continued treatment until disease progression, unacceptable adverse reactions, or other reasons for discontinuation.

[0485] Cohorts 1a, 2a, and 2c received, via subcutaneous (SC) infusion, a formulation of isatuximab containing 140 mg / mL, 9 mM histidine, 110 mM arginine monohydrochloride, 2 (w / v)% sucrose, and 0.4 (w / v)% poloxamer 188, pH 6.2. Cohorts 1a, 2a, and 2c additionally received 4 mg pomalidomide orally (po) on days 1 to 21 of every 28-day cycle; and 4 mg dexamethasone orally on days 1, 8, 15, and 22 of every 28-day cycle.

[0486] Cohorts 1b and 2b received a different formulation of isatuximab, pH 6.0, by intravenous (IV) infusion, containing 20 mg / mL isatuximab, 20 mM histidine, 10 (w / v)% sucrose, and 0.02 (w / v)% polysorbate 80. Cohorts 1b and 2b additionally received 4 mg pomalidomide orally (po) on days 1 to 21 of every 28-day cycle; and 4 mg dexamethasone orally on days 1, 8, 15, and 22 of every 28-day cycle.

[0487] Safety, PK, and RO data from cycle 1 (first 4 weeks) of cohort 1a and cohort 2a were reviewed before progression to cohort 2a or 2c, respectively.

[0488] Review of safety, PK, and RO data collected in cohorts 1(a / b) and 2(a / b) was used to support selection of the optimal SC isatuximab dose RP2D.

[0489] The 1000 mg isatuximab dose was selected as the starting dose based on PK modeling and simulations that demonstrated that, even assuming 80% SC bioavailability, isatuximab concentrations (day 28 trough plasma concentration [Ctrough]) were in a range similar to day 28 concentrations observed after IV dosing at 10 mg / kg and much lower IV doses of 20 mg / kg, which have been shown to be safe in the clinic. Doses lower than 1000 mg were not considered as starting doses due to the PK nonlinearity of isatuximab.

[0490] The selection of the 1400 mg dose of SC isatuximab was based on a population PK model constructed with IV data (n=127). This model demonstrated that, assuming an absolute bioavailability of 2:50%, 1400 mg of SC isatuximab administered QWx4 / Q2W would maintain Ctrough above the level achieved after 10 mg / kg IV QWx4 / Q2W. PK / PD analysis demonstrated that Ctrough at 4 weeks was a significant predictor of response (objective response rate, IV administration).

[0491] Patient inclusion criteria included: Patients previously diagnosed with multiple myeloma (MM) according to standard criteria and currently requiring treatment due to relapse of MM after response according to International Myeloma Working Group (IMWG) criteria.

[0492] Patients who received at least two prior therapies, including lenalidomide and a proteasome inhibitor, and who had disease progression during or after completion of the last therapy; and Patients with measurable disease defined as at least one of the following: · Serum M protein ≥ 0.5 g / dL (≥ 5 g / L); ≥ 200 mg / 24-hour urinary M protein; and · Serum free light chain (FLC) assay: FLC assay ≥ 10 mg / dL (≥ 100 mg / L) and abnormal serum FLC ratio (< 0.26 or > 1.65) were included.

[0493] Bone marrow and blood samples were collected for the following biomarker analyses: Isatuximab CD38 receptor occupancy will be measured on plasma cells from bone marrow aspirates and correlated with PK and clinical response parameters. Bone marrow samples were collected at screening and on Day 1 (pre-dose) of Cycle 2. This sample collection was stopped once the RP2D isatuximab SC dose was selected (Cohorts 1a / b and 2a / b only). Minimal residual disease (MRD) was investigated in bone marrow aspirates by next-generation sequencing and correlated with clinical response parameters. Bone marrow samples were collected for all participants at screening and at the time of maximal confirmed response of complete response (CR) or very good partial response (VGPR). Screening samples were only analyzed for participants achieving VGPR or higher. Using an isatuximab interference-removing assay, serum samples were investigated for potential isatuximab interference with M protein investigations in immunoelectrophoresis and immunofixation assays.

Claims

1. 1. A formulation comprising at least 100 mg / mL of an anti-CD38 antibody, The anti-CD38 antibody comprises a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences set forth in SEQ ID NOS: 1 to 3, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences set forth in SEQ ID NOS: 4 to 6, The formulation comprises a viscosity-reducing agent, a stabilizer, a buffer and a surfactant; The formulation has a pH of 5.9 to 7.0 and a viscosity of at most 25 mPa·s at 20°C.

2. 10. The formulation of claim 1, wherein the viscosity-reducing agent is 90-150 mM Arg-Cl.

3. 3. The formulation of claim 2, wherein the viscosity-reducing agent is 90-125 mM Arg-Cl.

4. 3. The formulation of claim 2, wherein the viscosity-reducing agent is 110 mM Arg-Cl.

5. 5. The formulation of any one of claims 1 to 4, wherein the surfactant is poloxamer 188.

6. 6. The formulation of any one of claims 1 to 5, wherein the surfactant is 0.4% (w / v) poloxamer 188.

7. The formulation of any one of claims 1 to 6, wherein the buffering agent is histidine.

8. The formulation of any one of claims 1 to 7, wherein the buffering agent is 9 mM histidine.

9. 10. The formulation of claim 1, wherein the viscosity-reducing agent is 90-150 mM Lys-Ac.

10. 10. The formulation of claim 9, wherein the viscosity-reducing agent is 125 mM Lys-Ac.

11. 11. The formulation of claim 9 or 10, wherein the surfactant is polysorbate 80.

12. 12. The formulation of claim 11, wherein the surfactant is 0.04 (w / v)% polysorbate 80.

13. The formulation of any one of claims 1 to 12, comprising 125 to 155 mg / mL of anti-CD38 antibody.

14. The formulation of any one of claims 1 to 13, comprising 140 mg / mL of anti-CD38 antibody.

15. The formulation of any one of claims 1 to 14, wherein the stabilizer is sucrose.

16. 16. The formulation of any one of claims 1 to 15, wherein the stabilizer is 2% (w / v) sucrose.

17. The formulation according to any one of claims 1 to 16, wherein the pH is from 5.9 to 7.

0.

18. 18. The formulation of claim 17, wherein the pH is 6.2 to 6.

3.

19. 1. A formulation comprising 140 mg / mL of an anti-CD38 antibody, The anti-CD38 antibody comprises a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences set forth in SEQ ID NOS: 1 to 3, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences set forth in SEQ ID NOS: 4 to 6, The formulation contained 9 mM histidine, 110 mM Arg-Cl, 2 (w / v) % sucrose and 0.4 (w / v) % poloxamer 188; The formulation has a pH of 6.2 to 6.3 and a viscosity of at most 14 mPa·s at 20°C.

20. 1. A formulation comprising 140 mg / mL of an anti-CD38 antibody, The anti-CD38 antibody comprises a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences set forth in SEQ ID NOS: 1 to 3, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences set forth in SEQ ID NOS: 4 to 6, The formulation contained 125 mM Lys-Ac, 2 (w / v)% sucrose and 0.04 (w / v)% polysorbate 80; The formulation has a pH of 6.2 and a viscosity of at most 14 mPa·s at 20°C.

21. A formulation according to any one of claims 1 to 20, which is suitable for subcutaneous administration.

22. 22. The formulation of claim 21, wherein the subcutaneous administration is a subcutaneous bolus administration.

23. The formulation of any one of claims 1 to 22, wherein the VH of the anti-CD38 antibody comprises the amino acid sequence shown in SEQ ID NO:7, and the VL of the anti-CD38 antibody comprises the amino acid sequence shown in SEQ ID NO:

8.

24. The formulation of any one of claims 1 to 23, wherein the anti-CD38 antibody is isatuximab.

25. A packaged pharmaceutical product comprising a sterile container containing a therapeutically effective amount of the formulation of any one of claims 1 to 24.

26. A device comprising a therapeutically effective amount of the formulation of any one of claims 1 to 24.

27. 27. The device of claim 26, selected from the group consisting of a syringe containing the formulation, a syringe driver, and an infusion pump.

28. 28. The device of claim 27, wherein the syringe is a pre-filled syringe.

29. CD38 ＋ 20. A method of treating a disease or condition characterized by the presence or activity of a cell, comprising administering to a subject in need thereof an effective amount of the formulation of any one of claims 1-8 or 13-19, wherein the formulation is administered subcutaneously.

30. CD38 ＋ The disease or condition characterized by the presence or activity of CD38 cells ＋ 30. The method of claim 29, wherein the tumor is a hematological malignancy.

31. CD38 ＋ 30. The method of claim 29, wherein the disease or condition characterized by the presence or activity of the cells is an autoimmune or inflammatory disease or condition.

32. CD38 ＋ 1. A method for treating hematological malignancies, comprising administering to a subject in need thereof a compound according to claim 1 to claim 6.

20. A method comprising administering an effective amount of the formulation of any one of claims 8 or 13-19, wherein the formulation is administered subcutaneously.

33. The formulation comprises 140 mg / mL of an anti-CD38 antibody; The anti-CD38 antibody comprises a variable heavy chain region (VH) comprising three complementarity determining regions (CDRs), CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences set forth in SEQ ID NOS: 1 to 3, and a variable light chain region (VL) comprising three CDRs, CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences set forth in SEQ ID NOS: 4 to 6, The formulation contained 9 mM histidine, 110 mM Arg-Cl, 2 (w / v) % sucrose and 0.4 (w / v) % poloxamer 188; 33. The method of claim 32, wherein the formulation has a pH of 6.2 or 6.3 and a viscosity of at most 14 mPa s at 20°C.

34. 34. The method of claim 32 or 33, wherein the formulation is administered by subcutaneous injection.

35. 35. The method of claim 34, wherein the subcutaneous injection is a bolus subcutaneous injection.

36. 35. The method of claim 34, wherein the subject is a human and the subcutaneous injection volume is 5 to 10 mL.

37. CD38 ＋ 35. The method of any one of claims 32 to 34, wherein the hematological malignancy is multiple myeloma.

38. 38. The method of claim 37, wherein the multiple myeloma is relapsed / refractory multiple myeloma.

39. 39. The method of claim 37 or 38, further comprising administering to the subject one or more agents selected from the group consisting of corticosteroids, proteasome inhibitors, immunomodulatory agents, chemotherapeutic agents, and combinations thereof.

40. 40. The method of claim 39, wherein the chemotherapeutic agent is selected from the group consisting of cytarabine, daunorubicin, daunomycin, doxorubicin, liposomal doxorubicin, idarubicin, mitoxantrone, gemtuzumab, clofarabine, cladribine, hydroxyurea, etoposide, melphalan, cyclophosphamide, vincristine, and combinations thereof.

41. 40. The method of claim 39, wherein the proteasome inhibitor is selected from the group consisting of carfilzomib, bortezomib, and combinations thereof.

42. 40. The method of claim 39, wherein the immunomodulatory agent is selected from the group consisting of thalidomide, lenalidomide, pomalidomide, and combinations thereof.

43. 40. The method of claim 39, wherein the corticosteroid is dexamethasone.

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