Dosage regimen and combination therapy for multispecific antibodies targeting B-cell maturation antigens

Abstract

The present disclosure relates to dosing regimens, formulations, and combinations comprising a multispecific antibody having binding specificity for at least a B cell maturation antigen (BCMA) and a T cell engaging arm; and methods of using such multispecific antibodies in the treatment or prevention of diseases, such as cancer.

Description

[0001] This application is a divisional application of PCT application PCT / IB2020 / 055872, filed on June 22, 2020, with the invention name “Dosage regimen and combination therapy for multispecific antibodies targeting B cell maturation antigens”. The date on which the PCT application entered the Chinese national phase is February 22, 2022, and the application number is 202080059452.0.

[0002] 1. Sequence Listing

[0003] This application contains a sequence listing that has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. The ASCII copy was created on May 14, 2020, is named NOV-011USP4_SL.txt, and is 15,390 bytes in size.

[0004] 2. Incorporation by Reference

[0005] All publications, patents, patent applications, and other documents cited in this application are incorporated herein by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, or other document was individually indicated to be incorporated by reference for all purposes. In the event of any inconsistency between one or more references incorporated herein and the teachings of the present disclosure, the teachings of the present specification are contemplated. Background Art

[0006] BCMA is a member of the tumor necrosis family receptor (TNFR) expressed on cells of the B cell lineage. BCMA expression is highest on terminally differentiated B cells (including plasma cells, plasmablasts, activated B cells, and subpopulations of memory B cells) with a long-lived plasma cell fate. BCMA participates in mediating the survival of plasma cells to maintain long-term humoral immunity. The expression of BCMA has been associated with a variety of cancers, autoimmune disorders, and infectious diseases. Cancers with increased BCMA expression include some hematological cancers, such as multiple myeloma, Hodgkin's lymphoma, and non-Hodgkin's lymphoma, various leukemias, and glioblastoma.

[0007] A variety of BCMA binding molecules are in clinical development, including BCMA antibody-drug conjugates, such as GSK2857916 (GlaxoSmithkline), and bispecific BCMA binding molecules targeting BMCA and CD3, such as PF06863135 (Pfizer), EM 901 (EngMab), JNJ-64007957 (Janssen), and AMG 420 (Amgen). See Cho et al., 2018, Front Immunol. [Immunology Frontier] 9: 1821; WO 2016 / 0166629.

[0008] One of the major safety concerns of any antibody-based drug, including CD3 bispecific molecules, is its potential to induce life-threatening side effects, such as cytokine release syndrome ("CRS"). See Shimabukuro-Vornhagen et al., 2018, J. Immunother Cancer. 6:56.

[0009] Therefore, there is an unmet medical need for polypeptides (eg, antibodies and multispecific binding molecules) that bind BCMA and have an improved safety profile (eg, reduced cytokine release) while still maintaining high efficacy.

[0010] Furthermore, there is an unmet medical need for appropriate dosing of antibodies and multispecific binding molecules that bind BCMA in order to reduce the chance of developing undesirable side effects, including CRS. Summary of the invention

[0011] Disclosed herein, inter alia, are methods of use, and formulations, combinations, and compositions comprising B cell maturation antigen (BCMA) binding molecules (e.g., multispecific antibodies, which may be immunoglobulin-based multispecific binding molecules (MBMs) described herein), more specifically BCMA binding molecules having the ability to target BCMA-expressing cells and the ability to engage T cells (e.g., by having a CD3 binding arm). The methods, formulations, combinations, and compositions are exemplified by BCMA binding molecules, referred to herein as BSBM3. Thus, references to "BCMA binding molecules" also apply to BSBM3.

[0012] Disclosed herein are methods of treating or preventing cancer (e.g., preventing the recurrence or recurrence of cancer), the methods comprising administering to a subject a BCMA-binding molecule at a dose of about 0.25 μg / kg to about 1200 μg / kg (e.g., 1 μg / kg to about 1000 μg / kg).

[0013] The BCMA binding molecules can be administered at different doses. For example, in one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 0.5 μg / kg to about 20 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 0.5 μg / kg to 10 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 1 μg / kg to 10 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 5 μg / kg to 10 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 1 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 3 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 6 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 10 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 10 μg / kg to 20 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 10 μg / kg to 15 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 12 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 20 μg / kg to about 40 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 20 μg / kg to about 30 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 24 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 30 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 40 μg / kg to about 80 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 40 μg / kg to about 60 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 48 μg / kg. In one embodiment, the BCMA-binding molecule is administered to a subject at a dose of about 80 μg / kg to about 120 μg / kg. In one embodiment, the BCMA-binding molecule is administered to a subject at a dose of about 80 μg / kg to about 100 μg / kg. In one embodiment, the BCMA-binding molecule is administered to a subject at a dose of about 96 μg / kg. In one embodiment, the BCMA-binding molecule is administered to a subject at a dose of about 100 μg / kg.In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 100 μg / kg to about 200 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 150 μg / kg to about 200 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 192 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 150 μg / kg to about 250 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 200 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 300 μg / kg to about 500 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 384 μg / kg. In one embodiment, the BCMA binding molecule is administered to a subject at a dose of about 400 μg / kg. In one embodiment, the BCMA-binding molecule is administered to a subject at a dose of about 500 μg / kg to about 700 μg / kg. In one embodiment, the BCMA-binding molecule is administered to a subject at a dose of about 600 μg / kg.

[0014] The BCMA-binding molecule can be administered to a subject in any effective manner. In some embodiments, the BCMA-binding molecule is administered intravenously to a subject.

[0015] If the BCMA-binding molecule can be administered intravenously to a subject, the BCMA-binding molecule is administered over a certain time span. For example, in one embodiment, the BCMA-binding molecule can be administered over a span of 2 hours.

[0016] The BCMA-binding molecule can also be administered to a subject once or more over the course of time. For example, in one embodiment, a BCMA-binding molecule can be administered to a subject once a week for four weeks.

[0017] The BCMA binding molecule can also be administered in an initial dose. The initial dose can be administered before starting treatment with a therapeutic dose (e.g., a therapeutically effective therapeutic dose). The initial dose can be a dose equal to or lower than the therapeutic dose subsequently administered. In some embodiments, the BCMA binding molecule is administered in an initial dose, and the initial dose is a dose lower than the first therapeutic dose. The initial dose can be administered in a single dose or in two or more doses. In some embodiments, the initial dose is divided into two administrations on two consecutive days. In some embodiments, one third of the initial dose is administered to the subject in one day, and two thirds of the initial dose is administered to the subject on the second day.

[0018] The BCMA-binding molecule can also be administered with a side effect reducer (e.g., acetaminophen and / or diphenhydramine). In one embodiment, the side effect reducer can be administered simultaneously with the BCMA-binding molecule. In one embodiment, the side effect reducer can be administered before the BCMA-binding molecule. In one embodiment, the side effect reducer can be administered after the BCMA-binding molecule.

[0019] The side effect reducer can reduce the onset or severity of cytokine release syndrome (CRS) in some cases. In one embodiment, the side effect reducer is a glucocorticoid. In one embodiment, the glucocorticoid is methylprednisolone. In one embodiment, methylprednisolone is given to the subject at a dosage of at least 2mg / kg. In one embodiment, the side effect reducer is acetaminophen, acetaminophen, antihistamines, steroids, anti-T cell directed therapy, or any combination thereof. In another embodiment, the side effect reducer is tocilizumab, canakinumab, or any combination thereof of anti-T cell directed therapy.

[0020] A subject who has received or will receive the BCMA-binding molecule may also be administered a second therapeutic agent. In some embodiments, the subject may receive one or more second therapeutic agents. In one embodiment, the BCMA-binding molecule and the second therapeutic agent are administered simultaneously, separately, or over a period of time.

[0021] In one embodiment, the second therapeutic agent is a gamma secretase inhibitor (GSI). In one embodiment, the GSI is LY-450139, PF-5212362, BMS-708163, MK-0752, ELN-318463, BMS-299897, LY-411575, DAPT, AL-101 (BMS-906024), AL-102 (BMS-986115), PF-3084014, RO4929097, or LY3039478. In one embodiment, the GSI is administered orally. In one embodiment, the GSI is administered prior to administering the BCMA-binding molecule.

[0022] In one embodiment, the second therapeutic agent is an immunomodulator. In one embodiment, the second therapeutic agent is an immune checkpoint inhibitor. In one embodiment, the second therapeutic agent is a TIM-3 inhibitor. In one embodiment, the TIM-3 inhibitor is MBG453. In one embodiment, the second therapeutic agent is a LAG-3 inhibitor. In one embodiment, the LAG-3 inhibitor is LAG525. In one embodiment, the second therapeutic agent is a PD-1 inhibitor. In one embodiment, the PD-1 inhibitor is PDR001, nivolumab, pembrolizumab, pidilizumab, MEDI0680, REGN2810, TSR-042, PF-06801591, BGB-A317, BGB-108, INCSHR1210, or AMP-224. In one embodiment, the PD-1 inhibitor is PDR001. In one embodiment, the PD-1 inhibitor is administered at a dose of about 100 mg once every four weeks, or about 200 mg once every four weeks, or about 300 mg once every four weeks, or about 400 mg once every four weeks, or about 500 mg once every four weeks. In one embodiment, the PD-1 inhibitor is administered at a dose of about 400 mg once every four weeks.

[0023] The second therapeutic agent can be administered in any effective manner. For example, the second therapeutic agent can be administered orally. In another embodiment, the second therapeutic agent can be administered intravenously.

[0024] The BCMA-binding molecule and / or the one or more second therapeutic agents can prevent or treat cancer. In one embodiment, the cancer is a blood cancer. In one embodiment, the blood cancer is multiple myeloma.

[0025] In some embodiments, the subject has previously been treated for cancer. In one embodiment, the subject suffers from recurrent and / or refractory multiple myeloma. In one embodiment, the subject has previously been treated with at least two prior treatment regimens. In one embodiment, the prior treatment regimen does not include a multispecific antibody. In one embodiment, the prior treatment regimen includes an immunomodulatory drug (IMiD), a proteasome inhibitor, an anti-CD38 inhibitor, or any combination thereof. In one embodiment, the prior treatment regimen includes an IMiD, and the IMiD is lenalidomide, pomalidomide, or both. In one embodiment, the prior treatment regimen includes a proteasome inhibitor, and the proteasome inhibitor is bortezomib, carfilzomib, or both. In one embodiment, the prior treatment regimen includes an anti-CD38 inhibitor, and the anti-CD38 inhibitor is an anti-CD38 antibody. In one embodiment, the anti-CD38 antibody is daratumumab. In one embodiment, the prior treatment regimen includes autologous bone marrow transplantation, BCMACAR-T, BCMA antibody-drug conjugate, or any combination thereof.

[0026] A subject treated with the BCMA-binding molecule may have (a) serum M-protein greater than or equal to 1.0 g / dL; (b) urine M-protein greater than or equal to 200 mg / 24 hours; (c) serum free light chains (sFLC) of affected FLC greater than 100 mg / L; or (d) any combination thereof.

[0027] In some embodiments, the subject treated with the BCMA-binding molecule is not eligible for treatment with other anti-cancer regimens known to provide clinical benefit.

[0028] Subjects treated with the BCMA-binding molecule may include the following: (a) no history of severe hypersensitivity reactions to the BCMA-binding molecule; (b) no history of toxicity from previous BCMA-targeted agents; (c) no other malignant diseases other than the cancer being treated and / or prevented; (d) no active, known or suspected autoimmune diseases; (e) no current treatment with contraindicated drugs that cannot be discontinued for at least one week prior to the start of treatment with the BCMA-binding molecule; (f) no infection with human immunodeficiency virus (HIV); V), active hepatitis B virus (HBV) or hepatitis C virus (HCV); (g) do not have impaired cardiac function or clinically significant cardiac disease including any of the following: (i) clinically significant and / or uncontrolled cardiac disease, such as congestive heart failure requiring treatment (NYHA class ≥2), uncontrolled hypertension, or clinically significant arrhythmias; (ii) QTcF>470msec on screening ECG or congenital long QT syndrome; or (iii) acute myeloid leukemia less than 3 months before the start of treatment with the BCMA-binding molecule. Myocardial infarction or unstable angina; (h) no radiation therapy within 14 days before the first dose of the BCMA-binding molecule, except for local radiation therapy for osteolytic lesions or plasmacytomas; (i) no major surgery within 2 weeks before the first dose of the BCMA-binding molecule; (j) no use of systemic chronic steroid therapy (≥10 mg / day of prednisone or equivalent), or any immunosuppressive therapy within 7 days of the first dose of the BCMA-binding molecule; (k) no systemic treatment with any immunosuppressive drugs; (l) no grade ≥2 neurological disease (m) plasma cell leukemia or other plasmacytoid disorders other than multiple myeloma; (n) not having any of the following clinical laboratory results: (i) absolute neutrophil count (ANC) <1,000 / mm3 without growth factor support within 7 days before the start of treatment; (ii) platelet count <75,000 mm3 without transfusion support within 7 days before the start of treatment; (iii) bilirubin >1.5% of the upper limit of normal range (ULN)5-fold; (iv) aspartate aminotransferase (AST) or alanine aminotransferase (ALT) > 3-fold ULN; or (v) creatinine clearance calculated by the Cockcroft-Gault equation < 30 ml / min; (o) no active infection requiring systemic therapy or other serious infection within 2 weeks prior to the first dose of the BCMA-binding molecule; (p) no POEMS syndrome (plasma cell dyscrasia with polyneuropathy, organomegaly, endocrinopathy, monoclonal protein, skin lesions, etc.) cell dyscrasia); (q) no prior allogeneic SCT at any time; (r) no use of any live vaccine against infectious diseases (e.g., influenza, varicella, pneumococcus) within 4 weeks of the first dose of the BCMA-binding molecule; (s) no treatment with cytotoxic antineoplastic drugs or small molecule targeted antineoplastic drugs or any experimental therapy within 14 days or 5 half-lives, which is shorter than the time before the first dose of the BCMA-binding molecule; (t) no induction of hematopoietic colony-stimulating growth factors (e.g., G-CSF, M-CSF), thrombopoietin mimetics or erythrocyte stimulating agents ≤ 2 weeks before the start of treatment; (u) no intravenous IG infusion for infection prevention within the last 28 days before treatment; (v) no presence of malignancy, or symptomatic CNS metastases or CNS metastases requiring local CNS-directed therapy (e.g., radiation therapy or surgery) within 2 weeks before the start of treatment, or (w) do not have a serious medical or psychiatric disorder that could interfere with treatment; (x) are not pregnant or lactating (lactating) women; (y) are not women of childbearing potential (defined as women who are physiologically capable of becoming pregnant) unless they are using an effective method of contraception (e.g., two methods) during dosing and for six months after the last dose of the BCMA-binding molecule, including at least one highly effective method of contraception, wherein highly effective methods of contraception include, but are not limited to, i) complete abstinence, ii) female sterilization, iii) male sterilization, and (iv) use of an oral, injectable, or implantable hormonal contraceptive method or placement of an intrauterine device (IUD) or intrauterine system (IUS) or other form of hormonal contraception with similar efficacy (failure rate <1%), such as a hormonal vaginal ring or transdermal hormonal contraception; and wherein the other effective method of contraception includes a barrier method of contraception, such as a condom or occlusive cap. cap) (diaphragm or cervical / vault cap) and a spermicide (e.g., foam, gel, film, cream or vaginal suppository); or (z) any combination thereof.

[0029] In some embodiments, administration of the BCMA-binding molecule is continued until the subject experiences toxicity, has clinical evidence of disease progression as determined by the IMWG, and / or treatment is discontinued at the discretion of the treating physician.

[0030] Further disclosed herein are combination therapies comprising a BCMA-binding molecule and a second therapeutic agent. In some embodiments, the combination therapy may comprise two or more second therapeutic agents.

[0031] In some embodiments, the second therapeutic agent is a gamma secretase inhibitor (GSI). In some embodiments, the GSI is LY-450139, PF-5212362, BMS-708163, MK-0752, ELN-318463, BMS-299897, LY-411575, DAPT, AL-101 (BMS-906024), AL-102 (BMS-986115), PF-3084014, RO4929097, or LY3039478.

[0032] In some embodiments, the second therapeutic agent is an immunomodulator. In some embodiments, the second therapeutic agent is an immune checkpoint inhibitor. In some embodiments, the second therapeutic agent is a TIM-3 inhibitor. In some embodiments, the TIM-3 inhibitor is MBG453. In some embodiments, the second therapeutic agent is a LAG-3 inhibitor. In some embodiments, the LAG-3 inhibitor is LAG525. In some embodiments, the second therapeutic agent is a PD-1 inhibitor. In some embodiments, the PD-1 inhibitor is PDR001, nivolumab, pembrolizumab, pidilizumab, MEDI0680, REGN2810, TSR-042, PF-06801591, BGB-A317, BGB-108, INCSHR1210, or AMP-224.

[0033] In some embodiments, the combination comprises about 100 mg, or about 200 mg, or about 300 mg, or about 400 mg, or about 500 mg of the second therapeutic agent. In some embodiments, the combination comprises about 2 mg, or about 10 mg, or about 20 mg, or about 40 mg, or about 80 mg, or about 160 mg, or about 320 mg of the compound; and about 100 mg, or about 200 mg, or about 300 mg, or about 400 mg, or about 500 mg of the second therapeutic agent.

[0034] Also described herein are combination therapies as disclosed for use in treating cancer. In some embodiments, the combination therapies are for use in preventing cancer.

[0035] In some embodiments, the use of the combination therapy as described herein is used to manufacture a medicament for treating or preventing cancer. In some embodiments, the use is for treating cancer. In some embodiments, the use is for preventing cancer.

[0036] In some embodiments, the cancer is a blood cancer. In some embodiments, the blood cancer is multiple myeloma.

[0037] Further described herein are pharmaceutical compositions comprising (a) a BCMA-binding molecule; (b) histidine; (c) sucrose; and (d) PS20.

[0038] In some embodiments, the composition is a liquid. In some embodiments, the histidine concentration is 20 mM. In some embodiments, the sucrose concentration is 240 mM. In some embodiments, the PS20 concentration is 0.04%. In some embodiments, the pH is about 5.5 ± 0.3.

[0039] Also described herein are vials comprising (a) 10 mg / mL of a BCMA-binding molecule; (b) 20 mM histidine; (c) 240 mM sucrose; (d) 0.04% PS20; and (e) a pH of about 5.5±0.3.

[0040] The present disclosure also discloses the following embodiments:

[0041] 1. A method of treating a subject having multiple myeloma, the method comprising administering to the subject one or more therapeutic doses of a bispecific antibody that binds to human BCMA and human CD3 and comprises:

[0042] (a) a first polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 1;

[0043] (b) a second polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 2; and

[0044] (c) a third polypeptide, wherein the amino acid sequence of the third polypeptide comprises the amino acid sequence of SEQ ID NO: 3.

[0045] 2. The method of embodiment 1, wherein the subject has a measurable disease.

[0046] 3. The method of embodiment 2, wherein the subject has a serum M-protein level ≥ 1 g / dL.

[0047] 4. The method according to embodiment 2 or embodiment 3, wherein the subject produces a urine M-protein level ≥ 200 mg / 24 hours.

[0048] 5. The method of any one of embodiments 2 to 4, wherein the subject has an affected FLC with a serum free light chain (sFLC) level of at least 100 mg / L.

[0049] 6. The method of any one of embodiments 1 to 5, wherein the multiple myeloma is relapsed.

[0050] 7. The method of any one of embodiments 1 to 6, wherein the multiple myeloma is refractory.

[0051] 8. The method of any one of embodiments 1 to 7, wherein the bispecific antibody is administered intravenously to the subject.

[0052] 9. The method of embodiment 8, wherein the bispecific antibody is administered to the subject as an infusion.

[0053] 10. The method of embodiment 9, wherein the infusion is over a span of 1.5-3 hours.

[0054] 11. The method of embodiment 9, wherein the infusion is over a span of 2 hours.

[0055] 12. The method of any one of embodiments 1 to 11, wherein one or more therapeutic doses are administered weekly.

[0056] 13. The method of any one of embodiments 1 to 12, wherein the subject receives at least three therapeutic doses.

[0057] 14. The method of embodiment 13, wherein the three therapeutic doses are administered over a period of less than one month.

[0058] 15. The method of embodiment 14, wherein the three therapeutic doses are administered over a period of 14 or 15 days.

[0059] 16. The method of any one of embodiments 1 to 12, wherein the subject receives at least four treatment doses.

[0060] 17. The method of embodiment 16, wherein the four therapeutic doses are administered over a period of less than one month.

[0061] 18. The method of embodiment 17, wherein the four therapeutic doses are administered over a period of 21, 22, or 23 days.

[0062] 19. The method of any one of embodiments 1 to 18, wherein one or more therapeutic doses or each therapeutic dose ranges from:

[0063] (a) from about 1 μg / kg to about 1200 μg / kg; or

[0064] (b) from about 50 μg to about 96 mg.

[0065] 20. The method of embodiment 19, wherein one or more of the therapeutic doses or each therapeutic dose ranges from:

[0066] (a) about 3 μg / kg to about 600 μg / kg; or

[0067] (b) from about 150 μg to about 48 mg.

[0068] 21. The method of embodiment 19, wherein one or more of the therapeutic doses or each therapeutic dose ranges from:

[0069] (a) about 5 μg / kg to about 100 μg / kg; or

[0070] (b) from about 150 μg to about 8 mg.

[0071] 22. The method of embodiment 19, wherein one or more of the therapeutic doses or each therapeutic dose ranges from:

[0072] (a) about 10 μg / kg to about 200 μg / kg; or

[0073] (b) from about 500 μg to about 16 mg.

[0074] 23. The method of embodiment 19, wherein one or more of the therapeutic doses or each therapeutic dose ranges from:

[0075] (a) about 50 μg / kg to about 400 μg / kg; or

[0076] (b) from about 2.5 mg to about 32 mg.

[0077] 24. The method of embodiment 19, wherein one or more of the therapeutic doses or each therapeutic dose ranges from:

[0078] (a) about 100 μg / kg to about 600 μg / kg; or

[0079] (b) from about 5 mg to about 96 mg.

[0080] 25. The method of embodiment 19, wherein the one or more therapeutic doses or the first therapeutic dose is:

[0081] (a) about 1 μg / kg; or

[0082] (b) from about 50 μg to about 80 μg.

[0083] 26. The method of embodiment 19, wherein the one or more therapeutic doses or the first therapeutic dose is:

[0084] (a) about 3 μg / kg; or

[0085] (b) from about 150 μg to about 240 μg.

[0086] 27. The method of embodiment 19, wherein the one or more therapeutic doses or the first therapeutic dose is:

[0087] (a) about 6 μg / kg; or

[0088] (b) from about 300 μg to about 480 μg.

[0089] 28. The method of embodiment 19, wherein the one or more therapeutic doses or the first therapeutic dose is:

[0090] (a) about 12 μg / kg; or

[0091] (b) from about 600 μg to about 960 μg.

[0092] 29. The method of embodiment 19, wherein the one or more therapeutic doses or the first therapeutic dose is:

[0093] (a) about 24 μg / kg; or

[0094] (b) from about 1.2 mg to about 1.92 mg.

[0095] 30. The method of embodiment 19, wherein the one or more therapeutic doses or the first therapeutic dose is:

[0096] (a) about 48 μg / kg; or

[0097] (b) from about 2.4 mg to about 3.84 mg.

[0098] 31. The method of embodiment 19, wherein the one or more therapeutic doses or the first therapeutic dose is:

[0099] (a) about 96 μg / kg; or

[0100] (b) from about 4.8 mg to about 7.68 mg.

[0101] 32. The method of embodiment 19, wherein the one or more therapeutic doses or the first therapeutic dose is:

[0102] (a) about 192 μg / kg; or

[0103] (b) from about 9.6 mg to about 15.36 mg.

[0104] 33. The method of embodiment 19, wherein the one or more therapeutic doses or the first therapeutic dose is:

[0105] (a) about 384 μg / kg; or

[0106] (b) from about 19.2 mg to about 30.72 mg.

[0107] 34. The method of embodiment 19, wherein the one or more therapeutic doses or the first therapeutic dose is:

[0108] (a) about 600 μg / kg; or

[0109] (b) from about 30 mg to about 48 mg.

[0110] 35. The method of any one of embodiments 1 to 34, wherein administering a therapeutic dose of the bispecific antibody comprises

[0111] (a) administering a first therapeutic dose; and

[0112] (b) escalating the therapeutic dose to a final therapeutic dose.

[0113] 36. The method of embodiment 35, comprising administering a second therapeutic dose that is the same as the first therapeutic dose prior to increasing the therapeutic dose.

[0114] 37. The method of embodiment 35 or embodiment 36, wherein step (b) comprises increasing the therapeutic dose more than once.

[0115] 38. The method of any one of embodiments 35 to 37, wherein each incremental dose results in no more than twice the previous dose.

[0116] 39. The method of any one of embodiments 35 to 38, wherein the first therapeutic dose ranges from:

[0117] (a) about 1 μg / kg to about 6 μg / kg; or

[0118] (b) from about 50 μg to about 480 μg.

[0119] 40. The method of any one of embodiments 35 to 38, wherein the first therapeutic dose is:

[0120] (a) about 1 μg / kg; or

[0121] (b) from about 50 μg to about 80 μg.

[0122] 41. The method of any one of embodiments 35 to 38, wherein the first therapeutic dose is:

[0123] (a) about 3 μg / kg; or

[0124] (b) from about 150 μg to about 240 μg.

[0125] 42. The method of any one of embodiments 35 to 38, wherein the first therapeutic dose is:

[0126] (a) about 6 μg / kg; or

[0127] (b) from about 300 μg to about 480 μg.

[0128] 43. The method of any one of embodiments 35 to 38, wherein the final therapeutic dose is in the range of:

[0129] (a) about 5 μg / kg to about 600 μg / kg; or

[0130] (b) from about 150 μg to about 48 mg.

[0131] 44. The method of any one of embodiments 35 to 38, wherein the final therapeutic dose is in the range of:

[0132] (a) about 10 μg / kg to about 200 μg / kg; or

[0133] (b) from about 500 μg to about 16 mg.

[0134] 45. The method of any one of embodiments 35 to 38, wherein the final therapeutic dose is in the range of:

[0135] (a) about 50 μg / kg to about 400 μg / kg; or

[0136] (b) from about 2.5 mg to about 32 mg.

[0137] 46. ​​The method of any one of embodiments 35 to 38, wherein the final therapeutic dose is in the range of:

[0138] (a) about 100 μg / kg to about 600 μg / kg; or

[0139] (b) from about 5 mg to about 96 mg.

[0140] 47. The method of any one of embodiments 35 to 38, wherein the final therapeutic dose is:

[0141] (a) about 6 μg / kg; or

[0142] (b) from about 300 μg to about 480 μg.

[0143] 48. The method of any one of embodiments 35 to 38, wherein the final therapeutic dose is:

[0144] (a) about 12 μg / kg; or

[0145] (b) from about 600 μg to about 960 μg.

[0146] 49. The method of any one of embodiments 35 to 38, wherein the final therapeutic dose is:

[0147] (a) about 24 μg / kg; or

[0148] (b) from about 1.2 mg to about 1.92 mg.

[0149] 50. The method of any one of embodiments 35 to 38, wherein the final therapeutic dose is:

[0150] (a) about 48 μg / kg; or

[0151] (b) from about 2.4 mg to about 3.84 mg.

[0152] 51. The method of any one of embodiments 35 to 38, wherein the final therapeutic dose is:

[0153] (a) about 96 μg / kg; or

[0154] (b) from about 4.8 mg to about 7.68 mg.

[0155] 52. The method of any one of embodiments 35 to 38, wherein the final therapeutic dose is:

[0156] (a) about 192 μg / kg; or

[0157] (b) from about 9.6 mg to about 15.36 mg.

[0158] 53. The method of any one of embodiments 35 to 38, wherein the final therapeutic dose is:

[0159] (a) about 384 μg / kg; or

[0160] (b) from about 19.2 mg to about 30.72 mg.

[0161] 54. The method of any one of embodiments 35 to 38, wherein the final therapeutic dose is:

[0162] (a) about 600 μg / kg; or

[0163] (b) from about 30 mg to about 48 mg.

[0164] 55. The method of any one of embodiments 1 to 54, comprising administering to the subject an initial dose of the bispecific antibody prior to administering a first therapeutic dose of the bispecific antibody.

[0165] 56. The method of embodiment 55, wherein the initial dose is lower than the first therapeutic dose.

[0166] 57. The method of embodiment 55, wherein the initial dose is equal to the first therapeutic dose.

[0167] 58. The method of any one of embodiments 55 to 57, wherein administration of the initial dose begins one week prior to administration of the first therapeutic dose.

[0168] 59. The method of embodiment 55 or embodiment 58, wherein the initial dose is divided.

[0169] 60. The method of embodiment 59, wherein the initial dose is administered over a period of two days.

[0170] 61. The method of embodiment 60, wherein less than half of the initial dose is administered on the first day and the remainder of the initial dose is administered on the second day.

[0171] 62. The method of embodiment 61, wherein about one third of the initial dose is administered on the first day and about two thirds of the initial dose is administered on the second day.

[0172] 63. The method of any one of embodiments 55 to 62, wherein the initial dose is in the range of:

[0173] (a) about 0.5 μg / kg to about 6 μg / kg; or

[0174] (b) from about 25 μg to about 480 μg.

[0175] 64. The method of embodiment 63, wherein the initial dose is:

[0176] (a) about 1 μg / kg; or

[0177] (b) from about 50 μg to about 80 μg.

[0178] 65. The method of embodiment 63, wherein the initial dose is:

[0179] (a) about 2 μg / kg; or

[0180] (b) from about 100 μg to about 160 μg.

[0181] 66. The method of embodiment 63, wherein the initial dose is:

[0182] (a) about 3 μg / kg; or

[0183] (b) from about 150 μg to about 240 μg.

[0184] 67. The method of embodiment 63, wherein the initial dose is:

[0185] (a) about 4 μg / kg; or

[0186] (b) from about 200 μg to about 320 μg.

[0187] 68. The method of embodiment 63, wherein the initial dose is:

[0188] (a) about 5 μg / kg; or

[0189] (b) from about 250 μg to about 400 μg.

[0190] 69. The method of embodiment 63, wherein the initial dose is:

[0191] (a) about 6 μg / kg; or

[0192] (b) from about 300 μg to about 480 μg.

[0193] 70. The method of any one of embodiments 1 to 34, comprising

[0194] (a) administering one third of the initial dose of the bispecific antibody to the subject on day 1 of treatment;

[0195] (b) on day 2 of treatment, administering to the subject two-thirds of the initial dose of the bispecific antibody;

[0196] (c) administering a first therapeutic dose to the subject on one of days 5 to 11 of treatment;

[0197] (d) administering a second therapeutic dose to the subject on one of days 12-18 of treatment; and

[0198] (e) administering a third therapeutic dose to the subject on one of days 19-25 of treatment.

[0199] 71. The method according to embodiment 70, comprising

[0200] (a) administering one third of the initial dose of the bispecific antibody to the subject on day 1 of treatment;

[0201] (b) on day 2 of treatment, administering to the subject two-thirds of the initial dose of the bispecific antibody;

[0202] (c) administering a first therapeutic dose to the subject on one of days 6 to 10 of treatment;

[0203] (d) administering a second therapeutic dose to the subject on one of days 13-17 of treatment; and

[0204] (e) administering a third therapeutic dose to the subject on one of days 20-24 of treatment.

[0205] 72. The method according to embodiment 70, comprising

[0206] (a) administering one third of the initial dose of the bispecific antibody to the subject on day 1 of treatment;

[0207] (b) on day 2 of treatment, administering to the subject two-thirds of the initial dose of the bispecific antibody;

[0208] (c) administering a first therapeutic dose to the subject on one of days 7 to 9 of treatment;

[0209] (d) administering a second therapeutic dose to the subject on one of days 14-16 of treatment; and

[0210] (e) administering a third therapeutic dose to the subject on one of days 21-23 of treatment.

[0211] 73. The method according to embodiment 70, comprising

[0212] (a) administering one third of the initial dose of the bispecific antibody to the subject on day 1 of treatment;

[0213] (b) on day 2 of treatment, administering to the subject two-thirds of the initial dose of the bispecific antibody;

[0214] (c) administering a first treatment dose to the subject on day 8 of treatment;

[0215] (d) administering a second treatment dose to the subject on day 15 of treatment; and

[0216] (e) administering a third treatment dose to the subject on day 22 of treatment.

[0217] 74. The method of any one of embodiments 70 to 73, wherein said initial dose is the same as said first therapeutic dose.

[0218] 75. The method of any one of embodiments 70 to 73, wherein said first therapeutic dose is 2 to 8 times greater than said initial dose.

[0219] 76. The method of any one of embodiments 70 to 73, wherein said first therapeutic dose is twice said initial dose.

[0220] 77. The method of any one of embodiments 70 to 73, wherein said first therapeutic dose is 4 times said initial dose.

[0221] 78. The method of any one of embodiments 70 to 73, wherein said first therapeutic dose is 8 times greater than said initial dose.

[0222] 79. The method of any one of embodiments 70 to 78, wherein the second therapeutic dose is equal to the first therapeutic dose.

[0223] 80. The method of any one of embodiments 70 to 79, wherein said third therapeutic dose is the same as said first therapeutic dose.

[0224] 81. The method of any one of embodiments 1 to 80, further comprising administering to the subject one or more agents that reduce side effects of the bispecific antibody.

[0225] 82. The method of embodiment 81, wherein the agent is administered before, simultaneously with, or after initiation of treatment with the bispecific antibody or any combination thereof.

[0226] 83. The method of embodiment 81 or embodiment 82, wherein the side effect is cytokine release syndrome (CRS).

[0227] 84. The method of embodiment 83, wherein the one or more agents reduce the onset or severity of CRS.

[0228] 85. The method of any one of embodiments 81 to 84, wherein said one or more agents comprises a glucocorticoid.

[0229] 86. The method of embodiment 85, wherein the glucocorticoid is methylprednisolone.

[0230] 87. The method of embodiment 86, wherein the methylprednisolone is administered at a dose of at least 2 mg / kg.

[0231] 88. The method of any one of embodiments 81 to 83, wherein the one or more agents comprise acetaminophen, paracetamol, an antihistamine, a steroid, an anti-T cell directed therapy, or any combination thereof.

[0232] 89. The method of embodiment 88, wherein the one or more agents comprise an anti-T cell directed therapy that is tocilizumab, canakinumab, or any combination thereof.

[0233] 90. The method of any one of embodiments 1 to 89, further comprising administering a second therapeutic agent to the subject.

[0234] 91. The method of embodiment 90, wherein the bispecific antibody and the second therapeutic agent are administered simultaneously, separately, or over a period of time.

[0235] 92. The method of embodiment 90 or 91, wherein the second therapeutic agent is a gamma secretase inhibitor (GSI).

[0236] 93. The method of embodiment 92, wherein the GSI is LY-450139, PF-5212362, BMS-708163, MK-0752, ELN-318463, BMS-299897, LY-411575, DAPT, AL-101 (BMS-906024), AL-102 (BMS-986115), PF-3084014, RO4929097, or LY3039478.

[0237] 94. The method of embodiment 93, wherein the GSI is AL-102.

[0238] 95. The method of any one of embodiments 92 to 94, wherein the GSI is administered orally.

[0239] 96. The method of any one of embodiments 92 to 95, wherein said GSI is administered prior to administration of said bispecific antibody.

[0240] 97. The method of embodiment 90 or 91, wherein the second therapeutic agent is an immunomodulator.

[0241] 98. The method of embodiment 90 or 91, wherein the second therapeutic agent is an immune checkpoint inhibitor.

[0242] 99. The method of embodiment 90 or 91, wherein the second therapeutic agent is a TIM-3 inhibitor.

[0243] 100. A method according to embodiment 99, wherein the TIM-3 inhibitor is MBG453.

[0244] 101. The method of embodiment 90 or 91, wherein the second therapeutic agent is a LAG-3 inhibitor.

[0245] 102. The method of embodiment 101, wherein the LAG-3 inhibitor is LAG525.

[0246] 103. The method of embodiment 90 or 91, wherein the second therapeutic agent is a PD-1 inhibitor.

[0247] 104. The method of embodiment 103, wherein the PD-1 inhibitor is PDR001, nivolumab, pembrolizumab, pidilizumab, MEDI0680, REGN2810, TSR-042, PF-06801591, BGB-A317, BGB-108, INCSHR1210, or AMP-224.

[0248] 105. The method of embodiment 103 or 104, wherein the PD-1 inhibitor is PDR001.

[0249] 106. The method of embodiment 104 or 105, wherein the PD-1 inhibitor is administered at a dose of about 100 mg once every four weeks, or about 200 mg once every four weeks, or about 300 mg once every four weeks, or about 400 mg once every four weeks, or about 500 mg once every four weeks.

[0250] 107. The method of embodiment 106, wherein the PD-1 inhibitor is administered at a dose of about 400 mg once every four weeks.

[0251] 108. The method of any one of embodiments 90 to 107, wherein the second therapeutic agent is administered intravenously.

[0252] 109. The method of any one of embodiments 1 to 108, wherein the subject has been previously treated with at least two prior treatment regimens.

[0253] 110. The method of embodiment 109, wherein the prior treatment regimen did not comprise a multispecific antibody, e.g., a bispecific antibody.

[0254] 111. The method of embodiment 109 or 110, wherein the prior treatment regimen comprises an immunomodulatory drug (IMiD), a proteasome inhibitor, an anti-CD38 inhibitor, or any combination thereof.

[0255] 112. The method of any one of embodiments 109 to 111, wherein the prior treatment regimen comprised an IMiD which is lenalidomide, pomalidomide, or both.

[0256] 113. The method of any one of embodiments 109 to 112, wherein the prior treatment regimen included a proteasome inhibitor, said proteasome inhibitor being bortezomib, carfilzomib, or both.

[0257] 114. The method of any one of embodiments 109 to 113, wherein the prior treatment regimen comprises an anti-CD38 inhibitor which is an anti-CD38 antibody.

[0258] 115. The method of embodiment 114, wherein the anti-CD38 antibody is daratumumab.

[0259] 116. The method of any one of embodiments 109 to 115, wherein the prior treatment regimen comprises autologous bone marrow transplantation, BCMA CAR-T, BCMA antibody-drug conjugate, or any combination thereof.

[0260] 117. The method of any one of embodiments 1 to 116, wherein the subject is:

[0261] (a) No history of severe hypersensitivity reaction to BSBM3;

[0262] (b) No history of toxicity from previous BCMA-targeted agents;

[0263] (c) not having any other malignant diseases except the cancer being treated and / or prevented;

[0264] (d) no active, known or suspected autoimmune disease;

[0265] (e) Currently receiving treatment with a prohibited drug that cannot be discontinued for at least one week prior to the start of the method;

[0266] (f) not infected with human immunodeficiency virus (HIV), active hepatitis B virus (HBV), or hepatitis C virus (HCV);

[0267] (g) Not have impaired cardiac function or clinically significant cardiac disease including any of the following:

[0268] (i) Clinically significant and / or uncontrolled cardiac disease, such as congestive heart failure (NYHA class ≥ 2) requiring treatment, uncontrolled hypertension, or clinically significant arrhythmias;

[0269] (ii) QTcF > 470 msec on screening ECG or congenital long QT syndrome; or

[0270] (iii) acute myocardial infarction or unstable angina <3 months before study entry;

[0271] (h) no radiotherapy within 14 days before the first dose, except for local radiotherapy for osteolytic lesions or plasmacytomas;

[0272] (i) No major surgery within 2 weeks before the first dose;

[0273] (j) not using chronic systemic steroid therapy (≥10 mg / day of prednisone or equivalent), or any immunosuppressive therapy within 7 days after the first dose;

[0274] (k) Not receiving any systemic treatment with immunosuppressive drugs;

[0275] (l) No neuropathy of grade ≥ 2 or residual toxic effects from previous therapy that have not resolved to grade ≤ 1 or baseline

[0276] (m) not suffering from plasma cell leukemia or other plasmacytoid disorders other than multiple myeloma;

[0277] (n) Does not have any of the following clinical laboratory results:

[0278] (i) absolute neutrophil count (ANC) <1,000 / mm3 without growth factor support within 7 days before the start of treatment;

[0279] (ii) platelet count <75,000 mm3 without transfusion support within 7 days before the start of treatment;

[0280] (iii) bilirubin >1.5 times the upper limit of normal range (ULN);

[0281] (iv) aspartate aminotransferase (AST) or alanine aminotransferase (ALT) > 3 times ULN; or

[0282] (v) creatinine clearance calculated according to the Cockcroft-Gault equation <30 ml / min;

[0283] (o) No active infection requiring systemic therapy or other serious infection within 2 weeks prior to the first dose;

[0284] (p) absence of POEMS syndrome (plasma cell dyscrasia with polyneuropathy, organomegaly, endocrinopathy, monoclonal protein, and skin lesions);

[0285] (q) no prior allogeneic SCT at any time; or

[0286] (r) not taking any live vaccines against infectious diseases (e.g. influenza, varicella, pneumococcal) within 4 weeks of the first dose;

[0287] (s) not treated with cytotoxic antineoplastic drugs or small molecule targeted antineoplastic drugs or any experimental therapy within 14 days or 5 half-lives, whichever is shorter than the time before the first dose;

[0288] (t) No hematopoietic colony-stimulating growth factors (e.g., G-CSF, M-CSF), thrombopoietin mimetics, or erythrocyte stimulating agents were initiated ≤ 2 weeks before the start of treatment;

[0289] (u) No intravenous IG infusion for infection prevention within the last 28 days before treatment;

[0290] (v) no active central nervous system (CNS) involvement due to malignancy, or the presence of symptomatic CNS metastases or CNS metastases requiring local CNS-directed therapy (e.g., radiotherapy or surgery), or escalation of the dose of corticosteroids within 2 weeks prior to the start of treatment;

[0291] (w) not suffering from a serious medical or mental illness that might interfere with treatment;

[0292] (x) if female, is not pregnant or lactating (lactating);

[0293] (y) if they are women of childbearing potential (defined as women who are physiologically capable of becoming pregnant), they use two effective methods of contraception during dosing and for 6 months after the last dose of study drug, with at least one of the effective methods of contraception being a highly effective method of contraception (e.g., i) complete abstinence, ii) female sterilization, iii) male sterilization, or (iv) use of an oral, injectable, or implantable hormonal method of contraception or placement of an intrauterine device (IUD) or intrauterine system (IUS) or other form of hormonal contraception with similar efficacy (failure rate <1%), such as a hormonal vaginal ring or transdermal hormonal contraception); or

[0294] (z) any combination thereof.

[0295] 118. The method of any one of embodiments 1 to 117, wherein the bispecific antibody is administered continuously until the subject experiences toxicity, has clinical evidence of disease progression as determined by IMWG, and / or treatment is discontinued at the discretion of the treating physician.

[0296] 119. The method of any one of embodiments 1 to 118, wherein the bispecific antibody is administered in the form of a pharmaceutical composition comprising:

[0297] (a) the bispecific antibody;

[0298] (b) histidine;

[0299] (c) sucrose; and

[0300] (d)PS20.

[0301] 120. The method of embodiment 119, wherein the pharmaceutical composition is in liquid form.

[0302] 121. The method of embodiment 119 or embodiment 120, wherein the concentration of histidine in the pharmaceutical composition is about 20 mM.

[0303] 122. The method of any one of embodiments 119 to 121, wherein the concentration of sucrose in the pharmaceutical composition is about 240 mM.

[0304] 123. The method of any one of embodiments 119 to 122, wherein the concentration of PS20 in the pharmaceutical composition is about 0.04%.

[0305] 124. The method of any one of embodiments 119 to 123, wherein the pH of said pharmaceutical composition is about 5.5±0.3.

[0306] 125. A vial, comprising:

[0307] (a) 10 mg / mL of a bispecific antibody that binds to human BCMA and human CD3 and comprises:

[0308] (i) a first polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 1;

[0309] (ii) a second polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 2; and

[0310] (iii) a third polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 3;

[0311] (b) 20 mM histidine;

[0312] (c) 240 mM sucrose;

[0313] (d) 0.04% PS20; and

[0314] (e) pH is about 5.5±0.3.

[0315] 126. A method of treating or preventing cancer, the method comprising administering to a subject in need thereof one or more therapeutic doses of a multispecific antibody having binding specificity for at least a B cell maturation antigen (BCMA) and a T cell engaging arm, wherein the multispecific antibody, optionally BSBM3, is administered to the subject at the following doses:

[0316] (a) about 1 μg / kg to about 1000 μg / kg;

[0317] (b) about 0.25 μg / kg to about 1200 μg / kg;

[0318] (c) from about 0.5 μg / kg to about 900 μg / kg; or

[0319] (d) from about 1 μg / kg to about 600 μg / kg.

[0320] 127. A method according to embodiment 126, wherein the T cell engaging arm binds to CD-3.

[0321] 128. The method of Embodiment 126 or Embodiment 127, wherein the multispecific antibody is administered to the subject at a dose of about 0.5 μg / kg to about 900 μg / kg.

[0322] 129. The method of any one of embodiments 126 to 127, wherein said multispecific antibody is administered to said subject at a dose of about 1 μg / kg to about 600 μg / kg.

[0323] 130. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 1 μg / kg.

[0324] 131. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 3 μg / kg.

[0325] 132. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 6 μg / kg.

[0326] 133. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 10 μg / kg.

[0327] 134. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 12 μg / kg.

[0328] 135. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 20 μg / kg to about 40 μg / kg.

[0329] 136. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 24 μg / kg.

[0330] 137. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 30 μg / kg.

[0331] 138. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 40 μg / kg to about 80 μg / kg.

[0332] 139. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 48 μg / kg.

[0333] 140. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 80 μg / kg to about 120 μg / kg.

[0334] 141. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 96 μg / kg.

[0335] 142. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 100 μg / kg.

[0336] 143. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 150 μg / kg to about 250 μg / kg.

[0337] 144. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 200 μg / kg.

[0338] 145. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 300 μg / kg to about 500 μg / kg.

[0339] 146. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 400 μg / kg.

[0340] 147. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 500 μg / kg to about 700 μg / kg.

[0341] 148. The method of Embodiment 127 or Embodiment 129, wherein the multispecific antibody is administered to the subject at a dose of about 600 μg / kg.

[0342] 149. The method of any one of embodiments 126 to 148, wherein said multispecific antibody is administered intravenously to said subject.

[0343] 150. The method of any one of embodiments 126 to 149, wherein the multispecific antibody is administered over a span of 2 hours.

[0344] 151. The method of any one of embodiments 126 to 150, wherein said multispecific antibody is administered to said subject once a week for 4 weeks.

[0345] 152. The method of any one of embodiments 126 to 151, wherein a primary dose is administered to the subject prior to administration of the first therapeutic dose.

[0346] 153. The method of embodiment 152, wherein the administration of the initial dose is divided.

[0347] 154. The method of embodiment 152 or 153, wherein the initial dose is administered to the subject over two days.

[0348] 155. The method of embodiment 154, wherein one third of the initial dose is administered on the first day and two thirds of the initial dose is administered on the second day.

[0349] 156. The method of embodiment 154 or embodiment 155, wherein the two days are consecutive.

[0350] 157. The method of any one of embodiments 152 to 156, wherein said initial dose is a dose lower than said therapeutic dose.

[0351] 158. The method of any one of embodiments 127 to 157, wherein a side effect reducing agent is administered to the subject.

[0352] 159. The method of embodiment 158, wherein the side effect reducer reduces the onset or severity of cytokine release syndrome (CRS).

[0353] 160. The method of embodiment 158 ​​or 159, wherein the side effect reducing agent is a glucocorticoid.

[0354] 161. The method of embodiment 160, wherein the glucocorticoid is methylprednisolone.

[0355] 162. The method of embodiment 161, wherein the methylprednisolone is administered at at least 2 mg / kg.

[0356] 163. The method of embodiment 158 ​​or 159, wherein the side effect reducer is acetaminophen, paracetamol, an antihistamine, a steroid, an anti-T cell directed therapy, or any combination thereof.

[0357] 164. The method of embodiment 163, wherein the side effect reducing agent is an anti-T cell directed therapy of tocilizumab, canakinumab, or any combination thereof.

[0358] 165. The method of any one of embodiments 126 to 164, wherein a second therapeutic agent is administered to the subject.

[0359] 166. The method of embodiment 165, wherein the multispecific antibody and the second therapeutic agent are administered simultaneously, separately, or over a period of time.

[0360] 167. The method of embodiment 165 or 166, wherein the second therapeutic agent is a gamma secretase inhibitor (GSI).

[0361] 168. The method of embodiment 167, wherein the GSI is LY-450139, PF-5212362, BMS-708163, MK-0752, ELN-318463, BMS-299897, LY-411575, DAPT, AL-101 (BMS-906024), AL-102 (BMS-986115), PF-3084014, RO4929097, or LY3039478.

[0362] 169. The method of embodiment 168, wherein the GSI is AL-102.

[0363] 170. The method of any one of embodiments 167 to 169, wherein the GSI is administered orally.

[0364] 171. The method of any one of embodiments 167 to 170, wherein said GSI is administered prior to administration of said multispecific antibody.

[0365] 172. The method of embodiment 165 or 166, wherein the second therapeutic agent is an immunomodulator.

[0366] 173. The method of embodiment 165 or 166, wherein the second therapeutic agent is an immune checkpoint inhibitor.

[0367] 174. The method of embodiment 165 or 166, wherein the second therapeutic agent is a TIM-3 inhibitor.

[0368] 175. A method according to embodiment 174, wherein the TIM-3 inhibitor is MBG453.

[0369] 176. The method of embodiment 165 or 166, wherein the second therapeutic agent is a LAG-3 inhibitor.

[0370] 177. The method of embodiment 176, wherein the LAG-3 inhibitor is LAG525.

[0371] 178. The method of embodiment 165 or 166, wherein the second therapeutic agent is a PD-1 inhibitor.

[0372] 179. The method of embodiment 178, wherein the PD-1 inhibitor is PDR001, nivolumab, pembrolizumab, pidilizumab, MEDI0680, REGN2810, TSR-042, PF-06801591, BGB-A317, BGB-108, INCSHR1210, or AMP-224.

[0373] 180. The method of embodiment 178 or 179, wherein the PD-1 inhibitor is PDR001.

[0374] 181. The method of embodiment 179 or 180, wherein the PD-1 inhibitor is administered at a dose of about 100 mg once every four weeks, or about 200 mg once every four weeks, or about 300 mg once every four weeks, or about 400 mg once every four weeks, or about 500 mg once every four weeks.

[0375] 182. The method of embodiment 181, wherein the PD-1 inhibitor is administered at a dose of about 400 mg once every four weeks.

[0376] 183. The method of any one of embodiments 165-182, wherein the second therapeutic agent is administered intravenously.

[0377] 184. The method of any one of embodiments 126 to 183, wherein the cancer is a hematological cancer.

[0378] 185. The method of embodiment 184, wherein the blood cancer is multiple myeloma.

[0379] 186. A method according to embodiment 184 or 185, wherein the subject has relapsed and / or refractory multiple myeloma.

[0380] 187. The method of any one of embodiments 126 to 186, wherein the subject has been previously treated with at least two prior treatment regimens.

[0381] 188. The method of embodiment 187, wherein the prior treatment regimen did not comprise a multispecific antibody.

[0382] 189. The method of embodiment 187 or 188, wherein the prior treatment regimen comprises an immunomodulatory drug (IMiD), a proteasome inhibitor, an anti-CD38 inhibitor, or any combination thereof.

[0383] 190. The method of any one of embodiments 187 to 189, wherein the prior treatment regimen comprised an IMiD which is lenalidomide, pomalidomide, or both.

[0384] 191. The method of any one of embodiments 187 to 190, wherein the prior treatment regimen included a proteasome inhibitor, said proteasome inhibitor being bortezomib, carfilzomib, or both.

[0385] 192. The method of any one of embodiments 187 to 191, wherein the prior treatment regimen comprises an anti-CD38 inhibitor which is an anti-CD38 antibody.

[0386] 193. The method of embodiment 192, wherein the anti-CD38 antibody is daratumumab.

[0387] 194. A method according to any one of embodiments 187 to 193, wherein the prior treatment regimen includes autologous bone marrow transplantation, BCMA CAR-T, BCMA antibody-drug conjugate, or any combination thereof.

[0388] 195. The method of any one of embodiments 126 to 194, wherein the subject has:

[0389] (a) serum M-protein greater than or equal to 1.0 g / dL;

[0390] (b) urinary M-protein greater than or equal to 200 mg / 24 hours;

[0391] (c) serum free light chains (sFLC) above 100 mg / L of affected FLC; or

[0392] (d) any combination thereof.

[0393] 196. The method of any one of embodiments 126 to 195, wherein the subject is:

[0394] (a) having no history of severe hypersensitivity reaction to the multispecific antibody;

[0395] (b) No history of toxicity from previous BCMA-targeted agents;

[0396] (c) not having any other malignant diseases except the cancer being treated and / or prevented;

[0397] (d) no active, known or suspected autoimmune disease;

[0398] (e) Currently receiving treatment with a prohibited drug that cannot be discontinued for at least one week prior to the start of the method;

[0399] (f) not infected with human immunodeficiency virus (HIV), active hepatitis B virus (HBV), or hepatitis C virus (HCV);

[0400] (g) Not have impaired cardiac function or clinically significant cardiac disease including any of the following:

[0401] (i) Clinically significant and / or uncontrolled cardiac disease, such as congestive heart failure (NYHA class ≥ 2) requiring treatment, uncontrolled hypertension, or clinically significant arrhythmias;

[0402] (ii) QTcF > 470 msec on screening ECG or congenital long QT syndrome; or

[0403] (iii) acute myocardial infarction or unstable angina <3 months before study entry;

[0404] (h) no radiotherapy within 14 days before the first dose, except for local radiotherapy for osteolytic lesions or plasmacytomas;

[0405] (i) No major surgery within 2 weeks before the first dose;

[0406] (j) not using chronic systemic steroid therapy (≥10 mg / day of prednisone or equivalent), or any immunosuppressive therapy within 7 days after the first dose;

[0407] (k) Not receiving any systemic treatment with immunosuppressive drugs;

[0408] (l) No neuropathy of grade ≥ 3 or residual toxic effects of grade ≥ 2 from previous therapy;

[0409] (m) not suffering from plasma cell leukemia or other plasmacytoid disorders other than multiple myeloma;

[0410] (n) Does not have any of the following clinical laboratory results:

[0411] (i) absolute neutrophil count (ANC) <1,000 / mm3 without growth factor support within 7 days before the start of treatment;

[0412] (ii) platelet count <75,000 mm3 without transfusion support within 7 days before the start of treatment;

[0413] (iii) bilirubin >1.5 times the upper limit of normal range (ULN);

[0414] (iv) aspartate aminotransferase (AST) or alanine aminotransferase (ALT) > 3 times ULN; or

[0415] (v) creatinine clearance calculated according to the Cockcroft-Gault equation <30 ml / min;

[0416] (o) No active infection requiring systemic therapy or other serious infection within 2 weeks prior to the first dose;

[0417] (p) absence of POEMS syndrome (plasma cell dyscrasia with polyneuropathy, organomegaly, endocrinopathy, monoclonal protein, and skin lesions);

[0418] (q) no prior allogeneic SCT at any time; or

[0419] (r) not taking any live vaccines against infectious diseases (e.g. influenza, varicella, pneumococcal) within 4 weeks of the first dose;

[0420] (s) not treated with cytotoxic antineoplastic drugs or small molecule targeted antineoplastic drugs or any experimental therapy within 14 days or 5 half-lives, whichever is shorter than the time before the first dose;

[0421] (t) No hematopoietic colony-stimulating growth factors (e.g., G-CSF, M-CSF), thrombopoietin mimetics, or erythrocyte stimulating agents were initiated ≤ 2 weeks before the start of treatment;

[0422] (u) No intravenous IG infusion for infection prevention within the last 28 days before treatment;

[0423] (v) no active central nervous system (CNS) involvement due to malignancy, or the presence of symptomatic CNS metastases or CNS metastases requiring local CNS-directed therapy (e.g., radiotherapy or surgery), or escalation of the dose of corticosteroids within 2 weeks prior to the start of treatment;

[0424] (w) not suffering from a serious medical or mental illness that might interfere with treatment;

[0425] (x) women who are not pregnant or breastfeeding;

[0426] (y) women of childbearing potential (defined as all women who are physiologically capable of becoming pregnant), unless they are using a highly effective method of contraception during dosing and for 90 days after the last dose of study drug, where such highly effective method of contraception includes i) complete abstinence, ii) female sterilization, iii) male sterilization, or (iv) use of an oral, injectable, or implantable hormonal method of contraception or placement of an intrauterine device (IUD) or intrauterine system (IUS) or other form of hormonal contraception with similar efficacy (failure rate <1%), such as a hormonal vaginal ring or transdermal hormonal contraception; or

[0427] (z) any combination thereof.

[0428] 197. The method of any one of embodiments 126 to 196, wherein the multispecific antibody is a bispecific antibody.

[0429] 198. The method of embodiment 197, wherein the bispecific antibody specifically binds to human BCMA and comprises the following: a first polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 1; a second polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 2; and a third polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 3.

[0430] 199. The method of any one of embodiments 126 to 198, wherein the multispecific antibody is administered continuously until the subject experiences toxicity, has clinical evidence of disease progression as determined by IMWG, and / or treatment is discontinued at the discretion of the treating physician.

[0431] 200. A combination therapy comprising a multispecific antibody having binding specificity for at least a B cell maturation antigen (BCMA) and a T cell engaging arm and a second therapeutic agent.

[0432] 201. The combination therapy according to embodiment 200, wherein the second therapeutic agent is a gamma secretase inhibitor (GSI).

[0433] 202. The combination therapy according to embodiment 201, wherein the GSI is LY-450139, PF-5212362, BMS-708163, MK-0752, ELN-318463, BMS-299897, LY-411575, DAPT, AL-101 (BMS-906024), AL-102 (BMS-986115), PF-3084014, RO4929097, or LY3039478.

[0434] 203. The combination therapy according to embodiment 202, wherein the GSI is AL-102.

[0435] 204. The combination therapy of embodiment 200, wherein the second therapeutic agent is an immunomodulator.

[0436] 205. The combination therapy according to embodiment 200, wherein the second therapeutic agent is an immune checkpoint inhibitor.

[0437] 206. The combination therapy according to embodiment 200, wherein the second therapeutic agent is a TIM-3 inhibitor.

[0438] 207. The combination therapy according to embodiment 206, wherein the TIM-3 inhibitor is MBG453.

[0439] 208. The combination therapy according to embodiment 200, wherein the second therapeutic agent is a LAG-3 inhibitor.

[0440] 209. The combination therapy of embodiment 208, wherein the LAG-3 inhibitor is LAG525.

[0441] 210. The combination therapy according to embodiment 200, wherein the second therapeutic agent is a PD-1 inhibitor.

[0442] 211. The combination therapy according to embodiment 210, wherein the PD-1 inhibitor is PDR001, nivolumab, pembrolizumab, pidilizumab, MEDI0680, REGN2810, TSR-042, PF-06801591, BGB-A317, BGB-108, INCSHR1210, or AMP-224.

[0443] 212. The combination therapy according to any one of embodiments 200 to 211, wherein the combination comprises about 100 mg, or about 200 mg, or about 300 mg, or about 400 mg, or about 500 mg of the second therapeutic agent.

[0444] 213. The combination therapy of any one of embodiments 200 to 211, wherein the combination comprises about 2 mg, or about 10 mg, or about 20 mg, or about 40 mg, or about 80 mg, or about 160 mg, or about 320 mg of the compound; and about 100 mg, or about 200 mg, or about 300 mg, or about 400 mg, or about 500 mg of the second therapeutic agent.

[0445] 214. The combination therapy according to any one of embodiments 200 to 213, for use in treating cancer.

[0446] 215. The combination therapy according to any one of embodiments 200 to 214, for use in preventing cancer.

[0447] 216. The combination therapy of embodiment 214 or embodiment 215, for administration according to the method of any one of embodiments 90 to 124 or 165 to 197.

[0448] 217. Use of the combination therapy according to any one of embodiments 200 to 216 for the manufacture of a medicament for the treatment or prevention of cancer.

[0449] 218. Use of the combination therapy according to any one of embodiments 200 to 216 for treating cancer.

[0450] 219. Use of the combination therapy according to any one of embodiments 200 to 216 for preventing cancer.

[0451] 220. The combination therapy according to any one of embodiments 200 to 216 or the use according to embodiments 217 to 219, wherein the cancer is a blood cancer.

[0452] 221. The combination therapy or use according to embodiment 220, wherein the blood cancer is multiple myeloma.

[0453] 222. A pharmaceutical composition comprising

[0454] (a) a multispecific antibody having binding specificity for at least a B cell maturation antigen (BCMA) and a T cell engaging arm;

[0455] (b) histidine;

[0456] (c) sucrose; and

[0457] (d)PS20.

[0458] 223. The pharmaceutical composition of embodiment 222, wherein said composition is a liquid.

[0459] 224. The pharmaceutical composition of embodiment 222 or 223, wherein the histidine concentration is 20 mM.

[0460] 225. The pharmaceutical composition of any one of embodiments 222 to 224, wherein the sucrose concentration is 240 mM.

[0461] 226. The pharmaceutical composition of any one of embodiments 222 to 225, wherein the PS20 concentration is 0.04%.

[0462] 227. The pharmaceutical composition of any one of embodiments 222 to 226, wherein the pH is about 5.5±0.3.

[0463] 228. A vial, comprising:

[0464] (a) 10 mg / mL of a multispecific antibody having binding specificity for at least a B cell maturation antigen (BCMA) and a T cell engaging arm

[0465] (b) 20 mM histidine;

[0466] (c) 240 mM sucrose;

[0467] (d) 0.04% PS20; and

[0468] (e) pH is about 5.5±0.3. BRIEF DESCRIPTION OF THE DRAWINGS

[0469] Figure 1 : The format of the BCMA-binding molecule called BSBM3.

[0470] Figures 2A-2C BSBM3-mediated T cell proliferation, cytokine production, and specific lysis of KMS11 myeloma cells via RTCC are shown. Healthy donor T cells were co-cultured with luciferase-overexpressing KMS11 cells at a 1:1 ratio in the presence of the indicated concentrations of BSBM3 or non-targeting (NT) control antibody. Figure 2A Shown are the levels of IFNγ and TNFα as measured by MSD assay using cell culture supernatants collected at 24 hr. Figure 2B Flow cytometry was used to show the + T cell counts were determined by event counting and normalized to counting bead controls after 4 days of co-culture. Figure 2C Shown is the %RTCC (% lysis of KMS11 cells) at 72 hr measured by reduction in luciferase activity compared to KMS11 cells alone. Shown are the mean + / - SEM of T cells from three independent healthy donors, each with three independent experiments (9 biological replicates in total).

[0471] Figure 3 Show that RTCC assay represents the most sensitive in vitro functional assay. For three different types of in vitro functional assays (RTCC, T cell proliferation and cytokine production), the EC30 values ​​of BSBM3 are plotted (as shown in Figure 2). Each data point represents one of nine biological replicates (T cells from three healthy donors were tested, each in three independent experiments).

[0472] Figure 4 Soluble BCMA is shown to reduce the activity of BSBM3 in the RTCC assay. The EC30 values ​​of BSBM3 in the RTCC assay with the addition of the soluble BCMA shown are shown. Each data point represents one of nine biological replicates (T cells from three healthy donor T cells were tested separately, each tested in three independent experiments).

[0473] Figure 5Antitumor activity of BSBM3 against KMS11 xenografts in a human PBMC adoptive transfer mouse model is shown. NSG mice were inoculated with KMS11 cells via tail vein injection on day 0 (D0), adoptively transferred with PBMCs on D7, and treated with BSBM3 at the following doses: 0.03 mg / kg (triangles), 0.3 mg / kg (circles), or 3.0 mg / kg (diamonds) on D15. For control: tumor-bearing mice without human PBMCs (increasing circles), tumor-bearing mice with human PBMCs but without Ab treatment (squares). Results from one representative experiment are shown from three biological replicates. *p<0.05, Dunnett's multiple comparison test. Data are expressed as geometric mean + / - SEM from 5 mice / group.

[0474] Figure 6 The clinical trial study protocol for BSBM3 is shown.

[0475] Figure 7 The international staging system for the BSBM3 clinical trial is shown.

[0476] Figure 8 The effect of AL-102 on BCMA shedding and membrane BCMA expression levels in KMS11 cells is shown. Soluble BCMA levels (ng / mL) from KMS11 cell culture supernatants treated with serially diluted AL-102 for 20 hours are shown on the left Y-axis. Antibody binding capacity (ABC) of anti-BCMA antibody (clone 19F2) on the surface of these AL-102-treated KMS11 cells is shown on the right Y-axis.

[0477] Fig. 9 BSBM3 EC50 (mean + / - SEM) is shown with increasing concentrations of AL-102: 1000 nM, which was diluted 5-fold over an 8-point serial. T cell donors n = 3. AL-102 enhanced the RTCC potency of BSBM3 in a dose-dependent manner. DETAILED DESCRIPTION

[0478] Although some embodiments have been / will be shown and described in the full text, such embodiments are provided only by way of example. Without departing from the present invention, multiple modifications, changes and replacements will be readily apparent to those skilled in the art. It should be understood that various alternatives to the embodiments of the present invention described herein will be adopted in practicing the present invention.

[0479] The present disclosure provides a method for treating and / or preventing a disease (e.g., cancer), comprising administering a composition comprising a BCMA-binding molecule (particularly a BCMA-binding molecule referred to as BSBM3) to a subject in need thereof. In some aspects, the method further comprises administering one or more therapeutic agents, such as one or more anti-tumor agents. The present disclosure further provides formulations, administration, dosing regimens and schedules, biomarkers, drug combinations, and other relevant clinical features.

[0480] According to the present disclosure, other therapeutic agents that can be used in combination with BCMA-binding molecules (e.g., BSBM3) are not limited to inhibitors of inhibitory molecules (e.g., checkpoint inhibitors), activators of costimulatory molecules, chemotherapeutic agents, targeted anticancer therapies, oncolytics, cytotoxic agents, or any therapeutic agents disclosed herein. In some embodiments, the one or more therapeutic agents can be PD-1 inhibitors, LAG-3 inhibitors, cytokines, A2A antagonists, GITR agonists, TIM-3 inhibitors, STING agonists, and TLR7 agonists for the treatment and / or prevention of patients / subjects with cancer.

[0481] The details of the present disclosure are set forth in the specification attached below. Although methods and materials similar or equivalent to those described herein can be used for the practice or testing of the present disclosure, illustrative methods and materials are now described. According to the specification and according to the claims, other features, objects and advantages of the present disclosure will be clear. In this specification and the appended claims, the singular form also includes the plural form, unless the context clearly indicates otherwise. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those of ordinary skill in the art to which the present disclosure belongs, which are generally understood.

[0482] 6.1. Definitions

[0483] As used herein, the following terms are intended to have the following meanings:

[0484] ADCC: As used herein, "ADCC" or "antibody-dependent cell-mediated cytotoxicity" refers to a cell-mediated reaction in which nonspecific cytotoxic cells expressing FcγRs recognize bound antibodies on target cells and subsequently cause lysis of the target cells. ADCC is associated with binding to FcγRIIIa; increased binding to FcγRIIIa results in increased ADCC activity.

[0485] ADCP : "ADCP" or antibody-dependent cell-mediated phagocytosis as used herein refers to a cell-mediated reaction in which nonspecific phagocytes expressing FcγRs recognize bound antibody on a target cell and subsequently cause phagocytosis of the target cell.

[0486] Other medicines : For convenience, agents used in combination with the antigen binding molecules of the present disclosure are referred to herein as "additional" agents.

[0487] Antibody : The term "antibody" as used herein refers to a polypeptide (or group of polypeptides) of the immunoglobulin family that can non-covalently, reversibly and specifically bind to an antigen. For example, a naturally occurring "antibody" of the IgG type is a tetramer comprising at least two heavy chains (H) and two light chains (L) interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region comprises three domains, namely CH1, CH2 and CH3. Each light chain comprises a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region consists of one domain (abbreviated herein as CL). The VH and VL regions can be further subdivided into hypervariable regions, called complementary determining regions (CDRs), interspersed with more conservative regions called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs arranged in the following order from the amino terminal to the carboxyl terminal: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant region of the antibody can mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system. The term "antibody" includes, but is not limited to, monoclonal antibodies, human antibodies, humanized antibodies, camelid antibodies, chimeric antibodies, bispecific or multispecific antibodies, and anti-idiotype (anti-Id) antibodies (including, for example, anti-Id antibodies for antibodies of the present disclosure). These antibodies may belong to any isotype / type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY) or subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2).

[0488] The light chain and the heavy chain are divided into structural homology regions and functional homology regions. The terms "constant" and "variable" are used functionally. In this regard, it should be understood that the variable domains of the light chain (VL) and the heavy chain (VH) portions determine antigen recognition and specificity. In contrast, the constant domains of the light chain (CL) and the heavy chain (CH1, CH2 or CH3) confer important biological properties such as secretion, transplacental mobility, Fc receptor binding, complement binding, etc. By convention, the farther the constant region domain is from the antigen binding site or amino terminus of the antibody, the larger its number. In wild-type antibodies, the variable region is at the N-terminus and the constant region is at the C-terminus; the CH3 domain and the CL domain actually contain the carboxyl termini of the heavy chain and the light chain, respectively.

[0489] Antibody fragments :As used herein, the term "antibody fragment" of an antibody refers to one or more parts of an antibody. In some embodiments, these parts are part of one or more contact domains of an antibody. In some other embodiments, these parts are antigen binding fragments (which retain the ability to bind to antigens non-covalently, reversibly and specifically), sometimes referred to herein as "antigen binding fragments", "antigen binding fragments thereof", "antigen binding portions", etc. Examples of binding fragments include, but are not limited to, single-chain Fv (scFv); Fab fragments; monovalent fragments consisting of VL, VH, CL and CH1 domains; F(ab)2 fragments, which are bivalent fragments comprising two Fab fragments connected by a disulfide bridge in the hinge region; Fd fragments consisting of VH and CH1 domains; Fv fragments consisting of the VL and VH domains of a single arm of an antibody; dAb fragments consisting of VH domains (Ward et al., 1989, Nature [Nature] 341: 544-546); and isolated complementary determining regions (CDRs). Thus, the term "antibody fragment" encompasses proteolytic fragments of an antibody (eg, Fab and F(ab)2 fragments) and engineered proteins comprising one or more portions of an antibody (eg, scFv).

[0490] Antibody fragments can also be incorporated into single domain antibodies, maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, v-NARs and bis-scFvs (see, e.g., Hollinger and Hudson, 2005 Nature Biotechnology 23: 1126-1136). Antibody fragments can be grafted into scaffolds based on polypeptides such as type III fibronectin (Fn3) (see U.S. Pat. No. 6,703,199, which describes fibronectin polypeptide monomers).

[0491] Antibody fragments can be incorporated into single-chain molecules comprising a pair of tandem Fv fragments (e.g., VH-CH1-VH-CH1), which together with complementary light chain polypeptides (e.g., VL-VC-VL-VC) form a pair of antigen binding regions (Zapata et al., 1995, Protein Eng. 8:1057-1062; and U.S. Pat. No. 5,641,870).

[0492] Antibody numbering system :In this specification, unless otherwise stated, reference to numbered amino acid residues in antibody domains is based on the EU numbering system. This system was originally designed by Edelman et al., 1969, Proc. Nat'l Acad. Sci. USA [Proceedings of the National Academy of Sciences of the United States] 63: 78-85 and by Kabat et al., 1991, in Sequences of Proteins of Immunological Interest [protein sequences with immunological importance], US Department of Health and Human Services (NIH, USA), the National Institutes of Health, USA, and described in detail.

[0493] Antigen binding domain : The term "antigen binding domain" or "ABD" refers to the portion of an antigen binding molecule that has the ability to bind to an antigen non-covalently, reversibly and specifically. Exemplary ABDs include antigen binding fragments and immunoglobulin-based and non-immunoglobulin-based scaffolds that retain the ability to bind to an antigen non-covalently, reversibly and specifically. As used herein, the term "antigen binding domain" encompasses antibody fragments that retain the ability to bind to an antigen non-covalently, reversibly and specifically.

[0494] Antigen binding domain chain or ABD chain :A single ABD may exist as one polypeptide chain (e.g., in the case of scFv) or be formed by the association of more than one polypeptide chain (e.g., in the case of Fab). As used herein, the term "ABD chain" refers to all or part of the ABD present on a single polypeptide chain. The use of the term "ABD chain" is intended for convenience and for descriptive purposes only, and does not imply a specific configuration or production method.

[0495] Fab : The term "antigen-binding fragment" of an antibody refers to a portion of an antibody that retains the ability to non-covalently, reversibly and specifically bind to an antigen.

[0496] Antigen Binding Molecules: The term "antigen binding molecule" refers to a molecule, such as an antibody, comprising one or more antigen binding domains. The antigen binding molecule may comprise one or more polypeptide chains, for example, one, two, three, four or more polypeptide chains. These polypeptide chains in an antigen binding molecule may be associated with each other directly or indirectly (for example, a first polypeptide chain may be associated with a second polypeptide chain, which in turn may be associated with a third polypeptide chain to form an antigen binding molecule, wherein the first and second polypeptide chains are directly associated with each other, the second and third polypeptide chains are directly associated with each other, and the first and third polypeptide chains are indirectly associated with each other through the second polypeptide chain).

[0497] Association : In the context of a domain or region in an antigen binding molecule, the term "association" refers to a functional relationship between two or more polypeptide chains and / or two or more parts of a single polypeptide chain. In particular, the term "association" means that two or more polypeptides (or parts of a single polypeptide) associate with each other, for example, non-covalently by molecular interactions and / or covalently by one or more disulfide bridges or chemical cross-links, thereby producing a functional antigen binding domain. Examples of associations that may exist in an antigen binding molecule include, but are not limited to, associations between Fc regions in an Fc domain, associations between VH and VL regions in a Fab or Fv, and associations between CH1 and CL in a Fab.

[0498] B cells: As used herein, the term "B cell" refers to a cell of the B cell lineage, which is a type of leukocyte of the lymphocyte subtype. Examples of B cells include plasmablasts, plasma cells, lymphoplasmacytoid cells, memory B cells, follicular B cells, marginal zone B cells, B-1 cells, B-2 cells, and regulatory B cells.

[0499] B-cell malignancies: As used herein, a B cell malignancy refers to the uncontrolled proliferation of B cells. Examples of B cell malignancies include non-Hodgkin's lymphoma (NHL), Hodgkin's lymphoma, leukemia, and myeloma. For example, the B-cell malignancy can be, but is not limited to, multiple myeloma, chronic lymphocytic leukemia (CLL) / small lymphocytic lymphoma (SLL), follicular lymphoma, mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), marginal zone lymphoma, Burkitt's lymphoma, lymphoplasmacytic lymphoma (Waldenstrom's macroglobulinemia), hairy cell leukemia, primary central nervous system (CNS) lymphoma, primary mediastinal large B-cell lymphoma, mediastinal gray zone lymphoma (MGZL), splenic marginal zone B-cell lymphoma, extranodal marginal zone B-cell lymphoma of MALT, nodal marginal zone B-cell lymphoma, and primary effusion lymphoma, and plasmacytoid dendritic cell neoplasm.

[0500] BCMA :As used herein, the term "BCMA" refers to B cell maturation antigen. BCMA (also known as TNFRSF17, BCMA or CD269) is a member of the tumor necrosis receptor (TNFR) family and is primarily expressed on terminally differentiated B cells (e.g., memory B cells and plasma cells). Its ligands include B cell activating factor (BAFF) and proliferation inducing ligand (APRIL). Protein BCMA is encoded by gene TNFRSF17. Exemplary BCMA sequences are available in the Uniprot database with accession number Q02223.

[0501] BCMA binding molecules : The term "BCMA-binding molecule" refers to a molecule that specifically binds to BCMA (particularly human BCMA). Examples of BCMA-binding molecules include multispecific binding molecules (which include at least one ABD that binds to BCMA), for example, multispecific antibodies, bispecific antibodies, and other bispecific binding molecules. The specific BCMA-binding molecule disclosed herein is referred to as BSBM3 herein.

[0502] Bispecific Binding Molecules : The term "bispecific binding molecule" or "BBM" refers to a molecule that specifically binds to two antigens and comprises two or more ABDs. The BBM of the present disclosure comprises at least one antigen binding domain that is specific for BCMA and at least one antigen binding domain that is specific for a different antigen (e.g., a component of a TCR complex). A BBM may comprise one, two, three, four, or even more polypeptide chains.

[0503] Two-price : In the context of antigen binding molecules, the term "bivalent" as used herein refers to an antigen binding molecule with two ABDs. The domains may be the same or different. Therefore, a bivalent antigen binding molecule may be monospecific or bispecific. A bivalent BBM comprises an ABD that specifically binds to BCMA and another ABD that binds to another antigen (e.g., a component of a TCR complex).

[0504] BSBM3 : BSMB3 refers to a BCMA-binding molecule comprising: (a) a first polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 1; (b) a second polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 2; and (c) a third polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 3. When co-expressed, the first, second and third polypeptides associate to form a Figure 1 Binding molecules in the configuration shown.

[0505] cancer : The term "cancer" refers to a disease characterized by the uncontrolled (and often rapid) growth of abnormal cells. Cancer cells can spread locally or to other parts of the body through the bloodstream and lymphatic system. Examples of various cancers are described herein and include, but are not limited to, leukemia, multiple myeloma, asymptomatic myeloma, Hodgkin's lymphoma, and non-Hodgkin's lymphoma, for example, any of the aforementioned BCMA-positive cancers of any type. The term "cancerous B cell" refers to a B cell that is undergoing or has undergone uncontrolled proliferation.

[0506] CD3 : The term "CD3" or "differentiation cluster 3" refers to the differentiation cluster 3 co-receptor of the T cell receptor. CD3 helps activate cytotoxic T cells (e.g., CD8+ naive T cells) and helper T cells (e.g., CD4+ naive T cells) and is composed of four different chains: a CD3γ chain (e.g., Genbank accession numbers NM_000073 and MP_000064 (human)), a CD3δ chain (e.g., Genbank accession numbers NM_000732, NM_001040651, NP_00732 and NP_001035741 (human)), and two CD3ε chains (e.g., Genbank accession numbers NM_000733 and NP_00724 (human)). The chains of CD3 are highly related cell surface proteins of the immunoglobulin superfamily containing a single extracellular immunoglobulin domain. The CD3 molecule associates with the T cell receptor (TCR) and the ζ chain to form a T cell receptor (TCR) complex, the role of which is to generate activation signals in T lymphocytes. Unless expressly stated otherwise, references to CD3 in this application may refer to CD3 co-receptors, CD3 co-receptor complexes, or any polypeptide chain of CD3 co-receptor complexes.

[0507] chimeric antibodies : The term "chimeric antibody" (or its antigen binding fragment) is an antibody molecule (or its antigen binding fragment) in which (a) the constant region or a portion thereof is altered, replaced or substituted so that the antigen binding site (variable region) is connected to a constant region of a different or altered type, effector function and / or kind, or to a completely different molecule (e.g., enzyme, toxin, hormone, growth factor, drug, etc.) that imparts new properties to the chimeric antibody; or (b) the variable region or a portion thereof is altered, replaced or substituted with a variable region having a different or altered antigen specificity. For example, a mouse antibody can be modified by replacing its constant region with a constant region from a human immunoglobulin. Due to replacement by a human constant region, the chimeric antibody can retain its specificity for recognizing an antigen while having reduced antigenicity in humans compared to the original mouse antibody.

[0508] Complementarity determining region :As used herein, the term "complementarity determining region" or "CDR" refers to the sequence of amino acids within the variable region of an antibody that imparts antigen specificity and binding affinity. For example, generally, there are three CDRs (e.g., CDR-H1, CDR-H2, and CDR-H3) in each heavy chain variable region, and there are three CDRs (CDR-L1, CDR-L2, and CDR-L3) in each light chain variable region. The precise amino acid sequence boundaries of a given CDR can be determined using any of a number of well-known schemes, including those described by Kabat et al., 1991, "Sequences of Proteins of Immunological Interest", 5th ed., National Institutes of Health, Public Health Service, Bethesda, Maryland ("Kabat" numbering scheme); Al-Lazikani et al., 1997 JMB 273, 927-948 ("Josiah" numbering scheme), or a combination thereof, and ImMunoGenTics (IMGT) numbering (Lefranc, 1999, The Immunologist, 7:132-136; Lefranc et al., 2003, Dev. Comp. Immunol. 27, 55-77 ("IMGT" numbering scheme). In the case of a given CDR region (e.g., HC CDR1, HC CDR2, HC CDR3, LC CDR4, LC CDR5, LC CDR6, LC CDR7, LC CDR8, LC CDR9, LC CDR10, LC CDR110, LC CDR120, LC CDR130, LC CDR140, LC CDR150, LC CDR160, LC CDR170, LC CDR180, LC CDR190, LC CDR210, LC CDR221 In some embodiments, the CDRs correspond to amino acid residues defined as part of a Kabat CDR, and amino acid residues defined as part of a Josiah CDR. As used herein, CDRs defined according to the "Josiah" numbering scheme are sometimes also referred to as "hypervariable loops."

[0509] For example, under Kabat numbering, the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 31-35 (CDR-H1) (e.g., one or more insertions after position 35), 50-65 (CDR-H2), and 95-102 (CDR-H3); and the CDR amino acid residues in the light chain variable domain (VL) are numbered 24-34 (CDR-L1) (e.g., one or more insertions after position 27), 50-56 (CDR-L2), and 89-97 (CDR-L3). As another example, under Josiah numbering, the CDR amino acid numbers in VH are 26-32 (CDR-H1) (e.g., one or more insertions after position 31), 52-56 (CDR-H2), and 95-102 (CDR-H3); and the amino acid residues in VL are 26-32 (CDR-L1) (e.g., one or more insertions after position 30), 50-52 (CDR-L2), and 91-96 (CDR-L3). By combining the CDR definitions of Kabat and Josiah, the CDRs include, for example, amino acid residues 26-35 (CDR-H1), 50-65 (CDR-H2), and 95-102 (CDR-H3) in human VH and amino acid residues 24-34 (CDR-L1), 50-56 (CDR-L2), and 89-97 (CDR-L3) in human VL or consist of them. According to IMGT, the CDR amino acid residues in VH are numbered approximately 26-35 (CDR1), 51-57 (CDR2), and 93-102 (CDR3), and the CDR amino acid residues in VL are numbered approximately 27-32 (CDR1), 50-52 (CDR2), and 89-97 (CDR3) (according to "Kabat" numbering). Under IMGT, the program IMGT / DomainGap Align can be used to determine the CDR regions of an antibody. In general, unless otherwise specified, the antibody molecule may include any combination of one or more Kabat CDRs and / or Josiah CDRs.

[0510] parallel : The term "concurrently" is not limited to administering therapies (e.g., prophylactic or therapeutic agents) at exactly the same time, but rather means that the pharmaceutical compositions comprising the antigen binding molecules are administered to a subject in a sequence and within a time interval such that the molecules can work together with one or more additional therapies to provide increased benefit (compared to if they were administered otherwise).

[0511] Conservative sequence modification: The term "conservative sequence modification" refers to an amino acid modification that does not significantly affect or change the binding characteristics of a BCMA binding molecule or its components (e.g., ABD or Fc region). Such conservative modifications include amino acid substitutions, additions, and deletions. Modifications can be introduced into BBM by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions are substitutions in which an amino acid residue is replaced by an amino acid residue with a similar side chain. Families of amino acid residues with similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), non-polar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, one or more amino acid residues in a BBM can be replaced with other amino acid residues from the same side chain family, and the altered BBM can be tested, for example, for binding to a target molecule and / or for effective heterodimerization and / or effector function.

[0512] Epitope : An epitope or antigenic determinant is a portion of an antigen that can be recognized by an antibody or other antigen binding moiety as described herein. An epitope can be linear or conformational.

[0513] Effector function: The term "effector function" refers to the activity of an antibody molecule, which is mediated by binding through the domain of the antibody rather than the antigen binding domain, usually mediated by the binding of an effector molecule. Effector functions include complement-mediated effector functions, which are mediated by the binding of, for example, the C1 component of the complement to the antibody. The activation of complement is important in the opsonization and lysis of cell pathogens. The activation of complement also stimulates inflammatory responses and can participate in autoimmune hypersensitivity reactions. Effector functions also include Fc receptor (FcR)-mediated effector functions, which can be triggered by the binding of the constant domain of the antibody to the Fc receptor (FcR). The binding of antibodies to Fc receptors on the cell surface triggers a large number of important and different biological responses, including phagocytosis and destruction of antibody-coated particles, clearance of immune complexes, ADCC, ADCP, release of inflammatory mediators, placental transport, and control of immunoglobulin production. The effector function of an antibody can be changed by, for example, enhancing or reducing the affinity of the antibody to effector molecules such as Fc receptors or complement components. Binding affinity will generally be altered by modifying the effector molecule binding site, and in this case it is appropriate to locate the site of interest and modify at least part of the site in a suitable manner. It is also envisioned that changes in the binding site on an antibody to an effector molecule need not significantly alter the overall binding affinity, but may alter the geometry of the interaction, resulting in ineffective effector mechanisms, as in nonproductive binding. It is further envisioned that effector function may also be altered by modifying sites that are not directly involved in effector molecule binding but that otherwise participate in the performance of effector function.

[0514] Fab :As used herein, "Fab" or "Fab region" refers to a polypeptide region comprising VH, CH1, VL, and CL immunoglobulin domains. These terms can refer to this region alone, or to this region in the context of an antigen binding molecule.

[0515] The Fab domain is formed by the association of the CH1 domain attached to the VH domain and the CL domain attached to the VL domain. The VH domain is paired with the VL domain to form the Fv region, and the CH1 domain is paired with the CL domain to further stabilize the binding module. The disulfide bond between the two constant domains can further stabilize the Fab domain.

[0516] Fab regions can be produced by proteolytic cleavage of immunoglobulin molecules (e.g., using enzymes such as papain) or by recombinant expression. In native immunoglobulin molecules, Fab is formed by the association of two different polypeptide chains (e.g., VH-CH1 on one chain associates with VL-CL on another chain). The Fab region is typically recombinantly expressed, typically on two polypeptide chains, although single-chain Fab is also contemplated herein.

[0517] Fc region :As used herein, the term "Fc region" or "Fc chain" refers to a polypeptide comprising the CH2-CH3 domain of an IgG molecule, and in some cases, includes a hinge. In the EU numbering of human IgG1, the CH2-CH3 domain includes amino acid 231 to amino acid 447, and the hinge is amino acid 216 to amino acid 230. Therefore, the definition of "Fc region" includes amino acid 231-447 (CH2-CH3) or 216-447 (hinge-CH2-CH3), or a fragment thereof. "Fc fragment" in this context may contain fewer amino acids from one or both of the N-terminal and C-terminal ends, but still retains the ability to form a dimer with another Fc region, which is detectable (e.g., non-denaturing chromatography, size exclusion chromatography) using standard methods (usually based on size). Human IgG Fc region has specific uses in the present disclosure, and may be an Fc region from human IgG1, IgG2, or IgG4.

[0518] Fc domain : The term "Fc domain" refers to a pair of associated Fc regions. The two Fc regions dimerize to produce an Fc domain. The two Fc regions in an Fc domain can be identical (such Fc domains are referred to herein as "Fc homodimers") or different from each other (such Fc domains are referred to herein as "Fc heterodimers").

[0519] F : The term "Fv", "Fv fragment" or "Fv region" refers to the region of the VL and VH domains of an antibody fragment that is in tight, non-covalent association (VH-VL dimer). In this configuration, the three CDRs of each variable domain interact to define the target binding site. Typically, six CDRs confer target binding specificity to the antigen binding molecule. However, in some cases, even a single variable domain (or half of an Fv that only contains three CDRs specific to the target) may have the ability to recognize and bind to the target. In natural immunoglobulin molecules, the VH and VL of Fv are located on separate polypeptide chains but can be engineered into single-chain Fv (scFv). The term also includes Fv engineered by introducing disulfide bonds for further stability.

[0520] Reference to VH-VL dimers herein is not intended to convey any particular configuration. For example, in a scFv, VH may be N-terminal or C-terminal to VL (the VH and VL are typically connected by a linker as discussed herein).

[0521] Half Antibody: The term "half antibody" refers to a molecule that comprises at least one ABD or one ABD chain and can associate with another molecule comprising an ABD or an ABD chain through, for example, a disulfide bridge or molecular interactions (e.g., knob-in-hole interactions between Fc heterodimers). A half antibody can be composed of one polypeptide chain or more than one polypeptide chain (e.g., two polypeptide chains of Fab). In one embodiment, a half antibody comprises an Fc region.

[0522] An example of a half antibody is a molecule comprising the heavy chain and light chain of an antibody (e.g., an IgG antibody). Another example of a half antibody is a molecule comprising a first polypeptide and a second polypeptide, wherein the first polypeptide comprises a VL domain and a CL domain, and the second polypeptide comprises a VH domain, a CH1 domain, a hinge domain, a CH2 domain, and a CH3 domain, wherein the VL and VH domains form an ABD. Yet another example of a half antibody is a polypeptide comprising a scFv domain, a CH2 domain, and a CH3 domain.

[0523] A half antibody may include more than one ABD, for example a half antibody comprising (in order from N-terminus to C-terminus) a scFv domain, a CH2 domain, a CH3 domain, and another scFv domain.

[0524] A half-antibody may also include an ABD chain that, when associated with another ABD chain in another half-antibody, forms a complete ABD.

[0525] Thus, a BBM may comprise one, more typically two, or even more than two half-antibodies, and a half-antibody may comprise one or more ABDs or one or more ABD chains.

[0526] In some BBMs, the first half antibody will associate with the second half antibody, e.g., heterodimerization. In other BBMs, the first half antibody will be covalently linked to the second half antibody, e.g., via a disulfide bridge or chemical cross-linking. In yet other BBMs, the first half antibody will associate with the second half antibody via covalent attachment and non-covalent interactions, e.g., disulfide bridges and knob-in-hole interactions.

[0527] The term "half antibody" is intended for descriptive purposes only and does not indicate a particular configuration or method of production. Descriptions of half antibodies as the "first" half antibody, the "second" half antibody, the "left" half antibody, the "right" half antibody, etc. are merely for convenience and descriptive purposes.

[0528] Hole: In the context of a knob-in-hole structure, a "hole" refers to at least one amino acid side chain that is recessed into the interface of a first Fc chain and can therefore be positioned in a complementary "knob" on the adjacent interfacing surface of a second Fc chain, thereby stabilizing the Fc heterodimer and thereby, for example, favoring the formation of Fc heterodimers over Fc homodimers.

[0529] Host cells or recombinant host cells : The term "host cell" or "recombinant host cell" refers to a cell that is genetically engineered, for example, by the introduction of a heterologous nucleic acid. It should be understood that this term refers not only to a specific subject cell, but also to the progeny of this cell. Because some changes may occur in the offspring due to mutations or environmental influences, this progeny may be different from the parent cell in fact, but is still included in the term "host cell" as used herein. The host cell can carry the heterologous nucleic acid transiently, for example, on an extrachromosomal heterologous expression vector, or stably carry the heterologous nucleic acid, for example, by integrating the heterologous nucleic acid into the host cell genome. For the purpose of expressing antigen binding molecules, the host cell can be a cell line of mammalian origin or a cell line with mammalian-like characteristics, such as monkey kidney cells (COS, for example, COS-1, COS-7), HEK293, baby hamster kidney (BHK, for example, BHK21), Chinese hamster ovary (CHO), NSO, PerC6, BSC-1, human hepatocellular carcinoma cells (for example, Hep G2), SP2 / 0, HeLa, Maddox bovine kidney (MDBK), myeloma and lymphoma cells or derivatives thereof and / or engineered variants. Engineered variants include, for example, modified and / or site-specific integration site derivatives of glycan profiles.

[0530] Humanization: The "humanized" form of the term non-human (e.g., mouse) antibody is a chimeric antibody containing the minimum sequence derived from a non-human immunoglobulin. In most cases, a humanized antibody is a human immunoglobulin (acceptor antibody), wherein the residues from the hypervariable region of the acceptor are replaced by the residues of the hypervariable region (donor antibody) with desired specificity, affinity and capacity from a non-human species (e.g., mouse, rat, rabbit or non-human primate). In some cases, the framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. In addition, the humanized antibody may be included in residues not found in the acceptor antibody or the donor antibody. These modifications are made to further improve antibody performance. Generally, a humanized antibody will include substantially all of the following items: at least one, typically two variable domains, wherein all or substantially all hypervariable loops correspond to those hypervariable loops of a non-human immunoglobulin, and all or substantially all FRs are those FRs of a human immunoglobulin sequence. The humanized antibody optionally also includes an immunoglobulin constant region (Fc), typically at least a portion of a human immunoglobulin constant region. Humanized antibodies are generally less immunogenic to humans than non-humanized antibodies and thus provide therapeutic benefits in certain circumstances. Humanized antibodies can be produced using known methods. See, for example, Hwang et al., 2005, Methods 36:35; Queen et al., 1989, Proc. Natl. Acad. Sci. USA 86:10029-10033; Jones et al., 1986, Nature 321:522-25, 1986; Riechmann et al., 1988, Nature 332:323-27; Ver Hoeyen et al., 1988, Science 239:1534-36; Orlandi et al., 1989, Proc. Natl. Acad. Sci. USA 86:3833-3837; U.S. Patent Nos. 5,225,539; 5,530,101; 5,585,089; 5,693,761; 5,693,762; and 6,180,370; and WO90 / 07861.See also the following review articles and references cited therein: Presta, 1992, Curr. Op. Struct. Biol. 2:593-596; Vaswani and Hamilton, 1998, Ann. Allergy, Asthma & Immunol. 1:105-115; Harris, 1995, Biochem. Soc. Transactions 23:1035-1038; Hurle and Gross, 1994, Curr. Op. Biotech. 5:428-433.

[0531] Human Antibodies :As used herein, the term "human antibody" includes antibodies having variable regions in which both the framework region and the CDR region are derived from sequences of human origin. In addition, if the antibody contains a constant region, the constant region is also derived from such human sequences, such as human germline sequences or mutant forms of human germline sequences, or antibodies containing consensus framework sequences derived from human framework sequence analysis, such as, for example, as described in Knappik et al., 2000. J Mol Biol [Molecular Biology] 296, 57-86. The structure and position of immunoglobulin variable domains (e.g., CDRs) can be defined using a well-known numbering scheme (e.g., the Kabat numbering scheme, the Chothia numbering scheme, or any combination of Kabat and Chothia) (see, e.g., Lazikani et al., 1997, J. Mol. Bio. 273:927-948; Kabat et al., 1991, Sequences of Proteins of Immunological Interest, 5th Edition, NIH Publication No. 91-3242 U.S. Department of Health and Human Services; Chothia et al., 1987, J. Mol. Biol. 196:901-917; Chothia et al., 1989, Nature 342:877-883).

[0532] Human antibodies may include amino acid residues not encoded by human sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro, or by somatic mutation in vivo, or conservative substitutions to promote stability or production). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species (e.g., a mouse) have been grafted into human framework sequences.

[0533] combination: As used herein, "combination" administration means that two (or more) different therapies are delivered to a subject during the subject's illness, for example, two or more therapies are delivered after the subject is diagnosed with a disorder and before the disorder is cured or cleared or before treatment is terminated for other reasons.

[0534] pestle( Knob ): In the context of a knob-in-hole structure, "knob" refers to at least one amino acid side chain that protrudes from the surface of a first Fc chain and can therefore be positioned in a complementary "hole" on the interface of a second Fc chain, thereby stabilizing the Fc heterodimer and thereby, for example, favoring the formation of Fc heterodimers over Fc homodimers.

[0535] Knobs and holes (or knobs-into-holes) One mechanism of Fc heterodimerization is often referred to in the art as "knobs and holes," or "knob-in-holes," or "knobs-into-holes." These terms refer to amino acid mutations that produce steric effects to favor the formation of Fc heterodimers over Fc homodimers, as described, for example, in Ridgway et al., 1996, Protein Engineering 9(7):617; Atwell et al., 1997, J. Mol. Biol. 270:26; and U.S. Pat. No. 8,216,805. Knobs and holes mutations can be combined with other strategies to improve heterodimerization.

[0536] Monoclonal antibodies As used herein, the term "monoclonal antibody" refers to polypeptides derived from the same genetic source, including antibodies, antibody fragments, molecules (including BBM), etc.

[0537] unit price : As used herein, in the context of antigen binding molecules, the term "monovalent" refers to an antigen binding molecule with a single antigen binding domain.

[0538] Multispecific Binding Molecules : The term "multispecific binding molecule" or "MBM" refers to an antigen binding molecule that specifically binds to at least two antigens and comprises two or more ABDs. The ABDs can each independently be an antibody fragment (e.g., scFv, Fab, nanobody), a ligand, or a non-antibody derived binder (e.g., fibronectin, Fynomer, DARPin).

[0539] Mutation or modification: In the context of the primary amino acid sequence of a polypeptide, the terms "modification" and "mutation" refer to amino acid substitutions, insertions, and / or deletions relative to a reference polypeptide polypeptide sequence. In addition, the term "modification" further encompasses changes to amino acid residues, such as by chemical conjugation (e.g., chemical conjugation of a drug or polyethylene glycol moiety) or post-translational modification (e.g., glycosylation).

[0540] Nucleic Acids : The term "nucleic acid" is used interchangeably with the term "polynucleotide" in this article, and refers to deoxyribonucleotides or ribonucleotides and polymers thereof in single-stranded or double-stranded form. The term encompasses nucleic acids containing known nucleotide analogs or modified backbone residues or connections, and nucleic acids are synthetic, naturally occurring and non-naturally occurring, have binding properties similar to reference nucleic acids, and are metabolized in a manner similar to reference nucleotides. Examples of such analogs include, but are not limited to, phosphorothioates, phosphoramidates, methylphosphonates, chiral-methylphosphonates, 2-O-methyl ribonucleotides and peptide-nucleic acids (PNAs).

[0541] Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions) and complementary sequences as well as explicitly indicated sequences. Specifically, as described in detail below, degenerate codon substitutions can be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed bases and / or deoxyinosine residues (Batzer et al., (1991) Nucleic Acid Res. 19:5081; Ohtsuka et al., (1985) J. Biol. Chem. 260:2605-2608; and Rossolini et al., (1994) Mol. Cell. Probes 8:91-98).

[0542] Operatively connected: The term "operably connected" refers to a functional relationship between two or more peptides or polypeptide domains or nucleic acid (e.g., DNA) segments. In the context of a fusion protein or other polypeptide, the term "operably connected" means connecting two or more amino acid segments to produce a functional polypeptide. For example, in the context of an antigen binding molecule, a separate ABM (or a chain of ABMs) can be operably connected via a peptide linker sequence. In the context of a nucleic acid encoding a fusion protein, such as a polypeptide chain of an antigen binding molecule, "operably connected" means connecting two nucleic acids so that the amino acid sequence encoded by the two nucleic acids remains in frame. In the context of transcriptional regulation, the term refers to the functional relationship between a transcriptional regulatory sequence and a transcribed sequence. For example, if a promoter or enhancer sequence stimulates or regulates the transcription of a coding sequence in an appropriate host cell or other expression system, the promoter or enhancer sequence is operably connected to the coding sequence.

[0543] Peptides and proteins : The terms "polypeptide" and "protein" are used interchangeably herein to refer to polymers of amino acid residues. The term encompasses amino acid polymers in which one or more amino acid residues are artificial chemical mimetics of the corresponding naturally occurring amino acids, as well as amino acid polymers applicable to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. In addition, the term encompasses amino acid polymers derivatized, for example, by synthetic derivatization of one or more side chains or ends, glycosylation, pegylation, circular permutations, cyclizations, attachment to linkers of other molecules, fusion to proteins or protein domains, and the addition of peptide tags or peptide labels.

[0544] Identification : As used herein, the term "recognizes" refers to an ABD that finds and interacts with (eg, binds to) its epitope.

[0545] Single-chain Fab or scFab : The terms "single-chain Fab" and "scFab" refer to a polypeptide comprising an antibody heavy chain variable domain (VH), an antibody constant domain 1 (CH1), an antibody light chain variable domain (VL), an antibody light chain constant domain (CL), and a linker such that the VH and VL are associated with each other and the CH1 and CL are associated with each other. In some embodiments, the antibody domains and linkers have one of the following orders in the direction from N-terminus to C-terminus: a) VH-CH1-linker-VL-CL, b) VL-CL-linker-VH-CH1, c) VH-CL-linker-VL-CH1 or d) VL-CH1-linker-VH-CL. The linker can be a polypeptide having at least 30 amino acids, e.g., between 32 and 50 amino acids. The single-chain Fab is stabilized by a natural disulfide bond between the CL domain and the CH1 domain.

[0546] Simultaneous or parallel delivery : In some embodiments, when the delivery of the second treatment begins, the delivery of the first treatment is still ongoing, so there is an overlap in terms of administration. This is sometimes referred to as "simultaneous delivery" or "concurrent delivery" in this article. In some embodiments of each case, the treatment is more effective due to the combined administration. For example, compared with the results observed when the second treatment is administered in the absence of the first treatment, the second treatment is more effective, such as an equivalent effect is observed with less of the second treatment, or the second treatment reduces the symptoms to a greater extent, or a similar situation is observed for the first treatment. In some embodiments, delivery reduces symptoms or other parameters related to the disorder more than the results observed when one treatment is delivered in the absence of the other. The effects of the two treatments can be partially additive, completely additive, or greater than additive. The delivery can be such that the effect of the first treatment delivered is still detectable when the second treatment is delivered.

[0547] Single-chain Fv or scFv : "Single-chain Fv" or "scFv" herein refers to a variable heavy domain covalently attached to a variable light domain, typically using an ABD linker as discussed herein, to form an scFv or scFv domain. The scFv domain can be in either direction from the N-terminus to the C-terminus (VH-linker-VL or VL-linker-VH). For a review of scFv, see Plückthun, in The Pharmacology of Monoclonal Antibodies [monoclonal antibody pharmacology], Vol. 113, Rosenburg and Moore, eds., (1994) Springer-Verlag [Springer], New York, pp. 269-315.

[0548] Bind specifically (or selectively) : The term "specifically (or selectively) binds" to an antigen or epitope refers to a binding reaction that determines the presence of a homologous antigen or epitope in a heterogeneous population of proteins and other biological agents. The antigen binding molecules or ABDs disclosed herein typically have a binding affinity of less than 5x10 -2 M, less than 10 -2 M, less than 5x10 -3 M, less than 10 -3 M, less than 5x10 -4 M, less than 10 -4 M, less than 5x10 -5 M, less than 10 -5 M, less than 5x10 -6 M, less than 10 -6 M, less than 5x10 -7 M, less than 10 - 7 M, less than 5x10-8 M, less than 10 -8 M, less than 5x10 -9 M, or less than 10 -9 The dissociation rate constant (KD) (koff / kon) of M is 1:1, and binds to the target antigen with an affinity at least two times (and more typically at least 20 times, at least 50 times, or at least 100 times) greater than its affinity for binding to a nonspecific antigen (e.g., HSA). The binding affinity can be measured using Biacore, SPR or BLI assays.

[0549] The term "specific binding" does not exclude cross-species reactivity. For example, an antigen binding moiety (e.g., an antigen binding fragment of an antibody) that "specifically binds" to an antigen from one species may also "specifically bind" to the antigen in one or more other species. Therefore, such cross-species reactivity itself does not change the classification of the antigen binding moiety as a "specific" binder. In certain embodiments, an antigen binding domain that specifically binds to a human antigen has cross-species reactivity with one or more non-human mammal species, e.g., primate species (including but not limited to one or more of cynomolgus monkeys (Macaca fascicularis), common macaques (Macaca mulatta), and pig-tailed monkeys (Macaca nemestrina)) or rodent species (e.g., mice (Mus musculus)). In other embodiments, the antigen binding domain does not have cross-species reactivity.

[0550] Subjects : The term "subject" includes humans and non-human animals. Non-human animals include all vertebrates, such as mammals and non-mammals such as non-human primates, sheep, dogs, cows, chickens, amphibians and reptiles. Unless otherwise indicated, the terms "patient" or "subject" are used interchangeably herein.

[0551] Tandem VH domains:As used herein, the term "tandem of VH domains (or VH)" refers to a string of VH domains, which consists of many identical VH domains of an antibody. The C-terminus of each of the VH domains (except the last one at the end of the tandem) is connected (with or without a linker) to the N-terminus of another VH domain. The tandem has at least 2 VH domains, and in some embodiments, the BBM has 3, 4, 5, 6, 7, 8, 9, or 10 VH domains. The tandem of VH can be produced by connecting the encoding nucleic acid of each VH domain in the desired order (this ensures that the nucleic acid is prepared as a single polypeptide chain) using a recombinant method (with or without a linker). The N-terminus of the first VH domain in the tandem is defined as the N-terminus of the tandem, and the C-terminus of the last VH domain in the tandem is defined as the C-terminus of the tandem.

[0552] Tandem VL domains :As used herein, the term "tandem of VL domains (or VL)" refers to a string of VL domains, which is composed of many identical VL domains of an antibody. The C-terminus of each VL domain (except the last one at the end of the series) is connected to (with or without a joint) the N-terminus of another VL. The series has at least 2 VL domains, and in some embodiments, the BBM has 3, 4, 5, 6, 7, 8, 9, or 10 VL domains. The series of VL can be produced by connecting the encoding nucleic acid of each VL domain (which ensures that the nucleic acid is prepared as a single polypeptide chain) in a desired order using a recombinant method (with or without a joint). The N-terminus of the first VL domain in the series is defined as the N-terminus of the series, and the C-terminus of the last VL domain in the series is defined as the C-terminus of the series.

[0553] Target Antigen : As used herein, "target antigen" refers to a molecule that is non-covalently, reversibly and specifically bound by an antigen binding domain.

[0554] Therapeutically effective amount : "Therapeutically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result.

[0555] Treatment:As used herein, the term "treatment (treat, treatment and treating)" refers to the progress of proliferative disorders due to the administration of one or more antigen binding molecules, the alleviation of severity and / or duration or improvement, or the improvement of one or more symptoms (e.g., one or more identifiable symptoms) of proliferative disorders. In certain embodiments, the term "treatment (treat, treatment and treating)" refers to improving at least one measurable physical parameter of proliferative disorders, such as the growth of tumors, which is not necessarily identifiable by patients. In other embodiments, the term "treatment (treat, treatment and treating)" refers to physically, or physiologically, or by both, suppressing the progress of proliferative disorders by, for example, stabilizing identifiable symptoms. In other embodiments, the term "treatment (treat, treatment and treating)" refers to reducing or stabilizing tumor size or cancer cell counts.

[0556] Tumor : The term "tumor" can be used interchangeably with the term "cancer" herein, e.g., both terms encompass solid and liquid tumors, such as diffuse or circulating tumors. As used herein, the term "cancer" or "tumor" includes pre-malignant and malignant cancers and tumors.

[0557] Variable region :As used herein, "variable region" or "variable domain" refers to the region of immunoglobulin, which comprises one or more Ig domains encoded by any one of Vκ, Vλ, and / or VH genes (these genes constitute κ, λ, and heavy chain immunoglobulin genetic loci, respectively) and contains CDRs that confer antigen specificity. "Variable heavy domain" can be paired with "variable light domain" to form antigen binding domain ("ABD"). In addition, each variable domain comprises three hypervariable regions ("complementarity determining regions", "CDR") (variable heavy domain is CDR-H1, CDR-H2, CDR-H3 and variable light domain is CDR-L1, CDR-L2, CDR-L3) and four framework regions (FR), which are arranged in the following order from amino terminal to carboxyl terminal: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4.

[0558] Carrier: The term "carrier" means a polynucleotide molecule capable of transporting another polynucleotide connected thereto. One type of carrier is a "plasmid", which refers to a circular double-stranded DNA loop, wherein additional DNA segments can be connected. Another type of carrier is a viral carrier, wherein additional DNA segments can be connected to the viral genome. Some carriers can replicate autonomously in the host cell into which they are introduced (e.g., bacterial carriers and additional mammalian carriers with bacterial replication origins). Other carriers (e.g., non-additional mammalian carriers) can be integrated into the genome of the host cell after being introduced into the host cell, thereby replicating together with the host genome. In addition, some carriers can guide the expression of genes operably connected thereto. Such carriers are referred to herein as "recombinant expression carriers" (or simply "expression carriers"). Generally, expression carriers useful in recombinant DNA technology are usually in the form of plasmids. In this specification, "plasmid" and "carrier" are used interchangeably because plasmids are the most commonly used carrier forms. However, the present disclosure is intended to include such other forms of expression vectors, such as viral vectors (eg, replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve the same functions.

[0559] VH : The term "VH" refers to the variable region of an immunoglobulin heavy chain of an antibody, including the heavy chain of Fv, scFv, dsFv or Fab.

[0560] V L : The term "VL" refers to the variable region of an immunoglobulin light chain (including the light chain of Fv, scFv, dsFv or Fab).

[0561] VH-VL or VH-VL pair : When referring to a VH-VL pair, whether located on the same polypeptide chain or different polypeptide chains, the terms "VH-VL" and "VH-VL pair" are used for convenience and are not intended to convey any particular orientation unless otherwise indicated by the context. Thus, a scFv comprising a "VH-VL" or a "VH-VL pair" may have the VH and VL domains in any orientation, for example, with the VH located at the N-terminus of the VL or the VL located at the N-terminus of the VH.

[0562] BCMA-binding molecules

[0563] The present disclosure provides treatment regimens and formulations of BCMA-binding molecules.Preferred BCMA-binding molecules are multispecific binding molecules (e.g., multispecific antibodies) that bind to BCMA and CD3, more specifically bispecific binding molecules (e.g., bispecific antibodies).

[0564] In a particularly preferred embodiment, the BCMA binding molecule is a molecule referred to herein as BSBM3. BSBM3 has a Fab domain targeting BCMA and a single-chain Fv (scFv) domain targeting CD3. BSBM3 consists of three polypeptides that, when expressed in the same cell, form Figure 1 Two half antibodies are shown. The first half antibody, consisting of a heavy chain polypeptide having an amino acid sequence of SEQ ID NO: 1 associated with a light chain polypeptide having an amino acid sequence of SEQ ID NO: 2, comprises a Fab domain that binds to CD19. The second half antibody, consisting of a heavy chain polypeptide having an amino acid sequence of SEQ ID NO: 3, comprises a scFv domain that binds to CD3. The Fc domain of BSBM3 comprises a substituent that eliminates binding to human Fcγ receptors in order to reduce the risk of non-selective T cell activation via FcR (Fc receptor)-mediated cross-linking. Without being bound by theory, it is believed that the Fc domain confers IgG-like in vivo persistence due to the affinity of unmodified FcRn (neonatal Fc receptor). It is also believed, without being bound by theory, that simultaneous binding of multiple molecules of BSBM3 to BCMA on multiple myeloma (MM) cells and the CD3 subunit of the T cell receptor (TCR) complex on T cells results in TCR cross-linking and the formation of cytolytic immune synapses, which leads to activation of T cells and specific lysis of MM cells.

[0565] 6.3. Pharmaceutical Compositions

[0566] The BCMA-binding molecules disclosed herein can be formulated as pharmaceutical compositions comprising the BCMA-binding molecules, for example, the pharmaceutical compositions contain one or more pharmaceutically acceptable excipients or carriers. In order to prepare a pharmaceutical or sterile composition comprising the BCMA-binding molecules disclosed herein, the BCMA-binding molecule preparation can be combined with one or more pharmaceutically acceptable excipients or carriers.

[0567] For example, formulations of BCMA-binding molecules can be prepared by mixing the BCMA-binding molecule with a physiologically acceptable carrier, excipient, or stabilizer in the form of, for example, a lyophilized powder, a slurry, an aqueous solution, a lotion, or a suspension (see, e.g., Hardman et al., 2001, Goodman and Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill, New York, NY; Gennaro, 2000, Remington: The Science and Practice of Pharmacy, Lippincott, Williams, and Wilkins, New York, NY; Avis et al. (eds.), 1993, Pharmaceutical Dosage Forms: General Medications, Marcel Dekker, NY; Dekker, NY; Lieberman et al. (eds.), 1990, Pharmaceutical Dosage Forms: Tablets, Marcel Dekker, NY; Lieberman et al. (eds.), 1990, Pharmaceutical Dosage Forms: Disperse Systems, Marcel Dekker, NY; Weiner and Kotkoskie, 2000, Excipient Toxicity and Safety, Marcel Dekker, Inc., NY).

[0568] For intravenous formulations, the BCMA-binding molecules can be formulated with one or more excipients. In one embodiment, the BCMA-binding molecules are formulated with amino acids. In one embodiment, the BCMA-binding molecules are formulated with sugars. In one embodiment, the BCMA-binding molecules are formulated with surfactants. In one embodiment, the BCMA-binding molecules are formulated with water. In some embodiments, the BCMA-binding molecules are formulated with one or more of amino acids, sugars, or surfactants.

[0569] In some embodiments, the amino acid may be histidine. In some embodiments, the sugar may be sucrose. In some embodiments, the surfactant may be a polysorbate, such as polysorbate 20 ("PS20"), also known as Tween 20.

[0570] Thus, the present disclosure provides a pharmaceutical composition comprising a BCMA-binding molecule and (a) an amino acid, such as histidine; (b) a sugar, such as sucrose; (c) a surfactant, such as PS20; or (d) a combination of any two or all of the foregoing. For parenteral (e.g., intravenous) administration, the pharmaceutical composition can be a liquid pharmaceutical composition.

[0571] Suitable concentrations of histidine range from 10 mM to 50 mM. In one embodiment, the concentration of histidine is 20 mM.

[0572] Suitable concentrations of sucrose range from 150 mM to 300 mM. In one embodiment, the concentration of sucrose is 240 mM.

[0573] Suitable concentrations of PS20 range from 0.02% to 0.06%. In one embodiment, the concentration of PS20 is 0.04%.

[0574] The pharmaceutical composition can be lyophilized and reconstituted in a suitable volume of liquid to obtain a solution for administration comprising, for example, one or more of histidine, sucrose, and PS20 in the above concentrations.

[0575] In some embodiments, the pH of the formulation can be acidic. For example, in one embodiment, the pH of the formulation can be about 5.0 to about 6.5. In one embodiment, the pH of the formulation can be about 5.0 to about 6.0. In some embodiments, the pH of the formulation can be about 5.5.

[0576] Suitable pH ranges for liquid pharmaceutical compositions comprising BCMA-binding molecules for parenteral (e.g., intravenous) administration are acidic, e.g., from about 5.0 to about 6.5. In certain aspects, the pH is from about 5.0 to about 6.0, and in some embodiments, the pH is about 5.5.

[0577] A suitable concentration range for the BCMA binding molecule is between 5 mg / mL and 20 mg / mL, and in one embodiment is 10 mg / mL.

[0578] Thus, in specific embodiments, the present disclosure provides a vial comprising (a) 10 mg / mL BSBM3; (b) 20 mM histidine; (c) 240 mM sucrose; (d) 0.04% PS20; and (e) a pH of about 5.5±0.3.

[0579] 7. Drug administration

[0580] 7.1. Amount of BCMA-binding molecules

[0581] BCMA-binding molecules can be used to prevent and / or treat cancer.

[0582] In some embodiments, the BCMA-binding molecule may be administered to a subject at about 0.5 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 1 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 10 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 30 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 50 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 75 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 100 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 200 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 300 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 400 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 500 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 600 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 700 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 800 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 900 μg / kg. In some embodiments, the BCMA-binding molecule may be administered to a subject at about 1000 μg / kg.

[0583] For example, in some embodiments, the BCMA binding molecule can be administered to a subject at a dose of from about 1 μg / kg to about 20 μg / kg. In some embodiments, the BCMA binding molecule can be administered to a subject at a dose of from about 20 μg / kg to about 40 μg / kg. In some embodiments, the BCMA binding molecule can be administered to a subject at a dose of from about 80 μg / kg to about 120 μg / kg. In some embodiments, the BCMA binding molecule can be administered to a subject at a dose of from about 150 μg / kg to about 250 μg / kg. In some embodiments, the BCMA binding molecule can be administered to a subject at a dose of from about 300 μg / kg to about 500 μg / kg. In some embodiments, the BCMA binding molecule can be administered to a subject at a dose of from about 500 μg / kg to about 700 μg / kg. In some embodiments, the BCMA binding molecule can be administered to a subject at a dose of from about 600 μg / kg to about 900 μg / kg.

[0584] In some embodiments, the BCMA binding molecule can be administered to a subject at a dose of from about 1 μg / kg to about 1000 μg / kg. In some embodiments, the BCMA binding molecule can be administered to a subject at a dose of from about 10 μg / kg to about 900 μg / kg. In some embodiments, the BCMA binding molecule can be administered to a subject at a dose of from about 20 μg / kg to about 800 μg / kg. In some embodiments, the BCMA binding molecule can be administered to a subject at a dose of from about 30 μg / kg to about 700 μg / kg. In some embodiments, the BCMA binding molecule can be administered to a subject at a dose of from about 50 μg / kg to about 600 μg / kg. In some embodiments, the BCMA binding molecule can be administered to a subject at a dose of from about 75 μg / kg to about 500 μg / kg. In some embodiments, the BCMA binding molecule can be administered to a subject at a dose of from about 100 μg / kg to about 400 μg / kg. In some embodiments, the BCMA-binding molecule can be administered to a subject at a dose of from about 150 μg / kg to about 300 μg / kg. In some embodiments, the BCMA-binding molecule can be administered to a subject at a dose of from about 200 μg / kg to about 250 μg / kg.

[0585] Furthermore, throughout this disclosure, any administration amount disclosed can be used to administer the BCMA-binding molecule, for example, as a first or subsequent therapeutic dose.

[0586] In one embodiment, the therapeutic dose may be about 1 μg / kg to about 1200 μg / kg or about 50 μg to about 96 mg. In another embodiment, the therapeutic dose may be about 3 μg / kg to about 600 μg / kg or about 150 μg to about 48 mg. In another embodiment, the therapeutic dose may be about 5 μg / kg to about 100 μg / kg or about 150 μg to about 8 mg. In another embodiment, the therapeutic dose may be about 10 μg / kg to about 200 μg / kg or about 500 μg to about 16 mg. In another embodiment, the therapeutic dose may be about 50 μg / kg to about 400 μg / kg or about 2.5 mg to about 32 mg. In another embodiment, the therapeutic dose may be about 100 μg / kg to about 600 μg / kg or about 5 mg to about 96 mg. In another embodiment, the therapeutic dose may be about 1 μg / kg or about 50 μg to about 80 μg. In another embodiment, the therapeutic dose may be about 3 μg / kg. In another embodiment, the therapeutic dose may be about 150 μg to about 240 μg. In another embodiment, the therapeutic dose may be about 6 μg / kg or about 300 μg to about 480 μg. In another embodiment, the therapeutic dose may be about 12 μg / kg or about 600 μg to about 960 μg. In another embodiment, the therapeutic dose may be about 24 μg / kg or about 1.2 mg to about 1.92 mg. In another embodiment, the therapeutic dose may be about 48 μg / kg or about 2.4 mg to about 3.84 mg. In another embodiment, the therapeutic dose may be about 96 μg / kg or about 4.8 mg to about 7.68 mg. In another embodiment, the therapeutic dose may be about 192 μg / kg or about 9.6 mg to about 15.36 mg. In another embodiment, the therapeutic dose may be about 384 μg / kg or about 19.2 mg to about 30.72 mg. In another embodiment, the therapeutic dose may be about 600 μg / kg or about 30 mg to about 48 mg.

[0587] In some cases, a first dose is needed or used. The first dose can be any dose described herein, and in some embodiments is lower than the first therapeutic dose. For example, if the first therapeutic dosage of the BCMA binding molecule is 30 μg / kg, the first dose can be given at any dose lower than 30 μg / kg. In this particular case, the first dose can be given at a dose lower than 30 μg / kg (e.g., lower than 29 μg / kg, such as 10 μg / kg or 1 μg / kg). In other embodiments, the first dose is equal to the first therapeutic dose. The first dose can be administered in a single administration or alternatively multiple administrations (e.g., twice). In some embodiments, one-third of the first dose is administered on the first day, and two-thirds of the first dose is administered on the second day (e.g., the day after the first day).

[0588] In one embodiment, the initial dose ranges from about 0.5 μg / kg to about 6 μg / kg or about 25 μg to about 480 μg. In another embodiment, the initial dose is about 1 μg / kg or about 50 μg to about 80 μg. In another embodiment, the initial dose is about 2 μg / kg or about 100 μg to about 160 μg. In another embodiment, the initial dose is about 3 μg / kg or about 150 μg to about 240 μg. In another embodiment, the initial dose is about 4 μg / kg or about 200 μg to about 320 μg. In another embodiment, the initial dose is about 5 μg / kg or about 250 μg to about 400 μg. In another embodiment, the initial dose is about 6 μg / kg or about 300 μg to about 480 μg.

[0589] 7.2. Time of administration

[0590] Administration can be performed for several hours. For example, if the administration of the BCMA binding molecule is performed intravenously, it can be accomplished by infusion. Infusion can occur in a time span of about 30 minutes to about 6 hours. In some embodiments, infusion can occur in a time span of about 30 minutes. In some embodiments, infusion can occur in a time span of about 1 hour. In some embodiments, infusion can occur in a time span of about 1.5 hours. In some embodiments, infusion can occur in a time span of about 2 hours. In some embodiments, infusion can occur in a time span of about 2.5 hours. In some embodiments, infusion can occur in a time span of about 3 hours. In some embodiments, infusion can occur in a time span of about 3.5 hours. In some embodiments, infusion can occur in a time span of about 4 hours. In some embodiments, infusion can occur in a time span of about 4.5 hours. In some embodiments, infusion can occur in a time span of about 5 hours. In some embodiments, infusion can occur in a time span of about 5.5 hours. In some embodiments, infusion can occur in a time span of about 6 hours.

[0591] In some embodiments, the infusion can occur in a time span of about 30 minutes to about 1 hour. In some embodiments, the infusion can occur in a time span of about 1 hour to about 2 hours. In some embodiments, the infusion can occur in a time span of about 2 hours to about 3 hours. In some embodiments, the infusion can occur in a time span of about 3 hours to about 4 hours. In some embodiments, the infusion can occur in a time span of about 4 hours to about 5 hours. In some embodiments, the infusion can occur in a time span of about 5 hours to about 6 hours.

[0592] In some embodiments, the infusion can occur in a time span of about 30 minutes to about 6 hours. In some embodiments, the infusion can occur in a time span of about 1 hour to about 5 hours. In some embodiments, the infusion can occur in a time span of about 1.5 hours to about 4 hours. In some embodiments, the infusion can occur in a time span of about 2 hours to about 3 hours.

[0593] In addition, any dosing time disclosed throughout can be used to administer the BCMA-binding molecules and / or any other therapeutic agents disclosed throughout.

[0594] 7.3. Dosage regimen

[0595] In some embodiments, the BCMA-binding molecule can be administered once a week. In some embodiments, the BCMA-binding molecule can be administered twice a week. In some embodiments, the BCMA-binding molecule can be administered once every two weeks.

[0596] In some embodiments, the BCMA-binding molecule can be administered in a single dose. In some embodiments, the BCMA-binding molecule can be administered in two doses. In some embodiments, the BCMA-binding molecule can be administered in three doses. In some embodiments, the BCMA-binding molecule can be administered in four doses.

[0597] In some embodiments, the BCMA-binding molecule can be administered for 1 week. In some embodiments, the BCMA-binding molecule can be administered for 2 weeks. In some embodiments, the BCMA-binding molecule can be administered for 3 weeks. In some embodiments, the BCMA-binding molecule can be administered for 4 weeks.

[0598] In some embodiments, the BCMA-binding molecules may be administered until remission (with respect to cancer), e.g., until a response is observed. In some embodiments, the BCMA-binding molecules may be administered until partial remission, e.g., until a partial response is observed. In some embodiments, the BCMA-binding molecules may be administered until complete remission, e.g., until a complete response is observed.

[0599] With respect to any given initial dose, the initial dose may be administered before the first therapeutic dose at any time prior to administering the therapeutic dose. For example, the initial dose may be administered once a week prior to administering the first therapeutic dose. In another example, the initial dose may be administered twice within a week prior to administering the first therapeutic dose.

[0600] In some embodiments, one third of the initial dose is administered to the subject on day 1 of the treatment course, and the remainder of the initial dose is administered on day 2 of the treatment. In some embodiments, the subject is then administered a first therapeutic dose on one of days 5-11 of the treatment (e.g., one of days 6-10, one of days 7-9, or day 8); the subject is then administered a second therapeutic dose on one of days 12-18 of the treatment (e.g., one of days 13-17, one of days 14-16, or day 15); and the subject is then administered a third therapeutic dose on one of days 19-25 of the treatment (e.g., one of days 20-24, one of days 21-23, or day 22).

[0601] 7.4. Side Effect Reducers

[0602] Regarding side effect reducing agents, the agents and dosages described throughout this disclosure can be used in the manner described throughout this disclosure. In addition, these side effect reducing agents can be used as known to be safe and effective.

[0603] 8. Combination

[0604] The BCMA binding molecules disclosed herein can be used in combination with other known agents and therapies. For example, the BCMA binding molecules can be used in a treatment regimen that is combined with surgery, chemotherapy, antibodies, radiation, peptide vaccines, steroids, cytotoxins, proteasome inhibitors, immunomodulatory drugs (e.g., IMiDs), BH3 mimetics, cytokine therapy, stem cell transplantation, or any combination thereof.

[0605] For convenience, agents used in combination with a BCMA-binding molecule are referred to herein as "additional" agents.

[0606] As used herein, "combination" administration means delivering two (or more) different treatments to a subject during the subject's illness, such as delivering two or more treatments after the subject is diagnosed with a disorder and before the disorder is cured or cleared or before treatment is terminated for other reasons. In some embodiments, delivery of the first treatment is still ongoing when delivery of the second treatment begins, so there is overlap in terms of administration. This is sometimes referred to as "simultaneous delivery" or "concurrent delivery" herein. The term "concurrent" is not limited to administering the therapy (e.g., BCMA-binding molecule and additional agent) at exactly the same time, but means administering a pharmaceutical composition comprising a BCMA-binding molecule to a subject in a sequence and within a certain time interval so that the BCMA-binding molecule can work together with one or more additional therapies to provide an increased benefit (compared to if they were administered otherwise). For example, each therapy can be administered to a subject at the same time or sequentially in any order at different time points; however, if not administered at the same time, they should be administered sufficiently close in time to provide the desired therapeutic effect.

[0607] The BCMA-binding molecule and one or more additional agents can be administered simultaneously or sequentially in the same or separate compositions. For sequential administration, the BCMA-binding molecule can be administered first and the additional agent can be administered secondly, or the order of administration can be reversed.

[0608] The BCMA-binding molecule and one or more additional agents can be administered to a subject in any suitable form and by any suitable route. In some embodiments, the routes of administration are the same. In other embodiments, the routes of administration are different.

[0609] In other embodiments, delivery of one treatment ends before delivery of the other treatment begins.

[0610] In some embodiments of each case, the treatment is more effective due to the combined administration. For example, the second treatment is more effective than the results observed when the second treatment is administered in the absence of the first treatment, such as an equivalent effect is observed with less of the second treatment, or the second treatment reduces symptoms to a greater extent, or similar conditions are observed for the first treatment. In some embodiments, delivery results in a greater reduction in symptoms or other parameters associated with the disorder than the results observed when the other treatment is delivered in the absence of one treatment. The effects of the two treatments may be partially additive, fully additive, or greater than additive. The delivery may be such that the effects of the first treatment delivered are still detectable when the second treatment is delivered.

[0611] The BCMA-binding molecule and / or additional agent can be administered during an active disorder, or during remission or less active disease. The BCMA-binding molecule can be administered prior to treatment with one or more additional agents, concurrently with treatment with one or more additional agents, after treatment with one or more additional agents, or during remission of the disorder.

[0612] When administered in combination, the BCMA-binding molecule and / or one or more additional agents can be administered in an amount or dosage that is higher, lower, or the same as the amount or dosage of each agent when used alone (e.g., as a monotherapy).

[0613] One or more additional agents of the combination therapy disclosed herein can be administered to the subject in parallel. The term "concurrently" is not limited to administering therapy (e.g., a prophylactic or therapeutic agent) at exactly the same time, but means administering a pharmaceutical composition comprising a BCMA-binding molecule to a subject in a sequence and at a certain time interval, so that the molecules disclosed herein can work together with one or more additional therapies to provide increased benefits (compared to when they are administered in other ways). For example, each therapy can be administered to the subject at the same time or in any order at different time points; however, if not administered at the same time, the therapy should be administered sufficiently close in time to provide the desired treatment or preventive effect. Each therapy can be administered to the subject in any appropriate form and by any suitable route.

[0614] The BCMA-binding molecule and one or more additional agents can be administered to a subject by the same or different routes of administration.

[0615] The BCMA-binding molecule and one or more additional agents can be administered in cycles. Cycling therapy involves administering a first therapy (e.g., a first prophylactic or therapeutic agent) for a period of time, then administering a second therapy (e.g., a second prophylactic or therapeutic agent) for a period of time, optionally, then administering a third therapy (e.g., a prophylactic or therapeutic agent) for a period of time, etc., and repeating this sequential administration, i.e., a cycle, to reduce the development of resistance to one of the therapies, to avoid or reduce the side effects of one of the therapies, and / or to improve the efficacy of the therapies.

[0616] In certain instances, the one or more additional agents are other anti-cancer agents, anti-allergic agents, anti-nausea agents (or anti-emetic agents), analgesics, cytoprotective agents, and combinations thereof.

[0617] The BCMA-binding molecules can be used in combination with a gamma secretase inhibitor ("GSI").

[0618] Thus, in one aspect, the disclosure provides a method for treating a subject having a disease associated with BCMA expression, the method comprising administering to the subject an effective amount of: (i) a BCMA-binding molecule, and (ii) a gamma secretase inhibitor (GSI).

[0619] In another aspect, the disclosure provides a method for treating a subject who has received treatment for a disease associated with BCMA expression, the method comprising administering to the subject an effective amount of: (i) a BCMA-binding molecule, and (ii) a GSI.

[0620] In one embodiment, the BCMA-binding molecule and the GSI are administered simultaneously or sequentially. In one embodiment, the BCMA-binding molecule is administered prior to the administration of the GSI. In one embodiment, the GSI is administered prior to the administration of the BCMA-binding molecule. In one embodiment, the BCMA-binding molecule and the GSI are administered simultaneously.

[0621] In one embodiment, the GSI is administered prior to administration of the BCMA-binding molecule (e.g., the GSI is administered 1, 2, 3, 4, or 5 days prior to administration of the BCMA-binding molecule), optionally wherein the subject exhibits an increase in cell surface BCMA expression levels and / or a decrease in soluble BCMA levels after administration of the GSI and prior to administration of the BCMA-binding molecule.

[0622] In some embodiments, the GSI is a small molecule that reduces the expression and / or function of γ-secretase, such as a small molecule GSI disclosed herein. In one embodiment, the GSI is selected from LY-450139, PF-5212362, BMS-708163, MK-0752, ELN-318463, BMS-299897, LY-411575, DAPT, AL-101 (also known as BMS-906024), AL-102 (also known as BMS-986115), PF-3084014, RO4929097, and LY3039478. In one embodiment, the GSI is selected from PF-5212362, ELN-318463, BMS-906024, and LY3039478. Exemplary GSIs are disclosed in Takebe et al., Pharmacol Ther. 2014 Feb;141(2):140-9; and Ran et al., EMBO Mol Med. 2017 Jul;9(7):950-966. In some embodiments, the GSI is AL-101. In some embodiments, the GSI is AL-102. The structure of AL-102 is shown below:

[0623]

[0624] In some embodiments, MK-0752 is administered in combination with docetaxel. In some embodiments, MK-0752 is administered in combination with gemcitabine. In some embodiments, BMS-906024 is administered in combination with chemotherapy.

[0625] In another embodiment, the GSI is a compound described in U.S. Pat. No. 7,468,365. In one embodiment, the GSI is LY-450139, i.e., semacid, (S)-2-hydroxy-3-methyl-N-((S)-1-(((S)-3-methyl-2-oxo-2,3,4,5-tetrahydro-1H-benzo[d]azepin-1-yl)amino)-1-oxopropan-2-yl)butanamide, or a pharmaceutically acceptable salt thereof. In one embodiment, the GSI is

[0626]

[0627] or a pharmaceutically acceptable salt thereof.

[0628] In another embodiment, the GSI is a compound described in U.S. Pat. No. 7,687,666. In one embodiment, the GSI is PF-5212362, i.e., begacestat, GSI-953, or (R)-5-chloro-N-(4,4,4-trifluoro-1-hydroxy-3-(trifluoromethyl)butan-2-yl)thiophene-2-sulfonamide, a pharmaceutically acceptable salt thereof. In one embodiment, the GSI is

[0629]

[0630] or a pharmaceutically acceptable salt thereof.

[0631] In another embodiment, the GSI is a compound described in U.S. Pat. No. 8,084,477. In one embodiment, the GSI is BMS-708163, i.e., avagacestat, or (R)-2-((4-chloro-N-(2-fluoro-4-(1,2,4-oxadiazol-3-yl)benzyl)phenyl)sulfonamide)-5,5,5-trifluoropentanamide, or a pharmaceutically acceptable salt thereof. In one embodiment, the GSI is

[0632]

[0633] or a pharmaceutically acceptable salt thereof.

[0634] In some embodiments, the GSI is a compound described in U.S. Pat. No. 7,160,875. In one embodiment, the GSI is RO4929097, i.e., (S)-2,2-dimethyl-N1-(6-oxo-6,7-dihydro-5H-dibenzo[b,d]azepin-7-yl)-N3-(2,2,3,3,3-pentafluoropropyl)malonamide, or a pharmaceutically acceptable salt thereof. In one embodiment, the GSI is

[0635]

[0636] or a pharmaceutically acceptable salt thereof.

[0637] In some embodiments, the GSI is

[0638]

[0639] or a pharmaceutically acceptable salt thereof.

[0640] In some embodiments, the GSI is a compound described in U.S. Pat. No. 6,984,663. In one embodiment, the GSI is MK-0752, i.e., 3-((1S,4R)-4-((4-chlorophenyl)sulfonyl)-4-(2,5-difluorophenyl)cyclohexyl)propanoic acid, or a pharmaceutically acceptable salt thereof. In some embodiments, the GSI is

[0641]

[0642] or a pharmaceutically acceptable salt thereof.

[0643] In some embodiments, the GSI is a compound described in U.S. Pat. No. 7,795,447. In one embodiment, the GSI is PF-3084014, i.e., nirogacestat or (S)-2-(((S)-6,8-difluoro-1,2,3,4-tetrahydronaphthyl-2-yl)amino)-N-(1-(2-methyl-1-(neopentylamino)propan-2-yl)-1H-imidazol-4-yl)pentanamide, or a pharmaceutically acceptable salt thereof.

[0644] In some embodiments, the GSI is

[0645]

[0646] or a pharmaceutically acceptable salt thereof.

[0647] In some embodiments, the GSI is a compound described in U.S. Pat. No. 7,939,657. In one embodiment, the GSI is ELN-318463, i.e., HY-50882 or (R)-N-(4-bromobenzyl)-4-chloro-N-(2-oxoazepan-3-yl)benzenesulfonamide, or a pharmaceutically acceptable salt thereof. In some embodiments, the GSI is

[0648]

[0649] or a pharmaceutically acceptable salt thereof.

[0650] In some embodiments, the GSI is a compound described in U.S. Pat. No. 8,629,136. In one embodiment, the GSI is BMS-906024, i.e., (2R, 3S)-N-[(3S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepine-3-yl]-2,3-bis(3,3,3-trifluoropropyl)succinamide, or a pharmaceutically acceptable salt thereof. In one embodiment, the GSI is

[0651]

[0652] or a pharmaceutically acceptable salt thereof.

[0653] In some embodiments, the GSI is a compound described in U.S. Pat. No. 8,629,136. In one embodiment, the GSI is LY3039478, i.e., crenigacestat or 4,4,4-trifluoro-N-((R)-1-(((S)-5-(2-hydroxyethyl)-6-oxo-6,7-dihydro-5H-benzo[d]pyrido[2,3-b]azepin-7-yl)amino)-1-oxopropan-2-yl)butanamide, or a pharmaceutically acceptable salt thereof. In some embodiments, the GSI is:

[0654]

[0655] or a pharmaceutically acceptable salt thereof.

[0656] In some embodiments, the GSI is BMS-299897, i.e., 2-[(1R)-1-[[(4-chlorophenyl)sulfonyl](2,5-difluorophenyl)amino]ethyl-5-fluorophenylbutyric acid or a pharmaceutically acceptable salt thereof. In some embodiments, the GSI is

[0657]

[0658] or a pharmaceutically acceptable salt thereof.

[0659] In some embodiments, the GSI is LY-411575, i.e., LSN-411575, (S)-2-((S)-2-(3,5-difluorophenyl)-2-hydroxyacetamido)-N-((S)-5-methyl-6-oxo-6,7-dihydro-5H-dibenzo[b,d]azepin-7-yl)propanamide, or a pharmaceutically acceptable salt thereof. In some embodiments, the GSI is

[0660]

[0661] or a pharmaceutically acceptable salt thereof.

[0662] In some embodiments, the GSI is DAPT, i.e., N-[(3,5-difluorophenyl)acetyl]-L-alanyl-2-phenyl]glycine-1,1-dimethylethyl ester or a pharmaceutically acceptable salt thereof. In some embodiments, the GSI is

[0663]

[0664] or a pharmaceutically acceptable salt thereof.

[0665] In some embodiments, the GSI is a compound described in U.S. Patent Publication No. US-2015-307533 (e.g., in the table on pages 13-16). In some embodiments, the GSI is selected from:

[0666]

[0667] or a pharmaceutically acceptable salt thereof.

[0668] In some embodiments, the GSI is a compound of U.S. Pat. No. 8,188,069. In one embodiment, the GSI is

[0669] or a pharmaceutically acceptable salt thereof.

[0670] In some embodiments, the GSI is a compound described in U.S. Pat. No. 9,096,582 (e.g., in the table on pages 13-17). In some embodiments, the GSI is selected from:

[0671]

[0672] or a pharmaceutically acceptable salt thereof.

[0673] In some embodiments, the GSI is a compound described in U.S. Patent Publication No. US-2011-0257163 (e.g., in paragraphs

[0506] to

[0553] ). In some embodiments, the GSI is selected from:

[0674]

[0675]

[0676] and pharmaceutically acceptable salts thereof.

[0677] In some embodiments, the GSI is selected from:

[0678]

[0679] and pharmaceutically acceptable salts thereof. In some embodiments, the GSI is selected from:

[0680]

[0681]

[0682] and pharmaceutically acceptable salts thereof.

[0683] In some embodiments, the GSI is an antibody molecule that reduces the expression and / or function of γ-secretase. In some embodiments, the GSI is an antibody molecule that targets a subunit of γ-secretase. In some embodiments, the GSI is selected from an anti-presenilin antibody molecule, an anti-nicastrin antibody molecule, an anti-APH-1 antibody molecule, or an anti-PEN-2 antibody molecule.

[0684] Exemplary antibody molecules targeting a gamma-secretase subunit (eg, for example, presenilin, desenelin, APH-1, or PEN-2) are described in US 8,394,376, US 8,637,274, and US 5,942,400.

[0685] In one aspect, the present disclosure provides a method for treating a subject with a B cell disorder or disorder, the method comprising administering to the subject an effective amount of: (i) a BCMA binding molecule, and (ii) a gamma secretase modulator (e.g., a GSI). Exemplary B cell disorders or disorders that can be treated with a combination of a BCMA binding molecule and a gamma secretase modulator include multiple myeloma, Waldenstrom's macroglobulinemia, chronic lymphocytic leukemia, B cell non-Hodgkin's lymphoma, plasmacytoma, Hodgkin's lymphoma, follicular lymphoma, non-cleaved small cell lymphoma, endemic Burkitt's lymphoma, sporadic Burkitt's lymphoma, marginal zone lymphoma, extranodal mucosa-associated lymphoid tissue lymphoma, nodular monocytic B cell lymphoma, splenic lymphoma, mantle cell lymphoma, large cell lymphoma, diffuse mixed cell lymphoma, immunoblastic lymphoma, primary mediastinal B cell lymphoma, pulmonary B cell angiocentric lymphoma, Lymphoma, small lymphocytic lymphoma, B-cell proliferation of uncertain malignant potential, lymphomatoid granulomatosis, post-transplant lymphoproliferative disorder, immunoregulatory disorders, rheumatoid arthritis, myasthenia gravis, idiopathic thrombocytopenic purpura, antiphospholipid syndrome, Chagas disease, Graves disease, Wegener's granulomatosis, polyarteritis nodosa, Sjögren's syndrome, pemphigus vulgaris, scleroderma, multiple sclerosis, antiphospholipid syndrome, ANCA-associated vasculitis, Goodpasture's disease, Kawasaki disease, autoimmune hemolytic anemia, rapidly progressive glomerulonephritis, heavy chain disease, primary or immune cell-associated amyloidosis, and monoclonal gammopathy of undetermined significance.

[0686] In some embodiments, the γ-secretase modulator is a γ-secretase modulator described in WO 2017 / 019496. In some embodiments, the γ-secretase modulator is γ-secretase inhibitor I (GSI I) Z-Leu-Leu-norleucine; γ-secretase inhibitor II (GSI II); γ-secretase inhibitor III (GSI III), N-benzyloxycarbonyl-Leu-leucine, N-(2-naphthoyl)-Val-phenylalaninal; γ-secretase inhibitor IV (GSI IV); γ-secretase inhibitor V (GSI V), N-benzyloxycarbonyl-Leu-phenylalaninal; γ-secretase inhibitor VI (GSI VI), 1-(S)-endo-N-(1,3,3)-trimethylbicyclo[2.2.1]hept-2-yl)-4-fluorophenylsulfonamide; γ-secretase inhibitor VII (GSI VII), menthyloxycarbonyl-LL-CHO; γ-secretase inhibitor IX (GSI IX), (DAPT), N-[N-(3,5-difluorophenylacetyl-L-alanyl)]-S-phenylglycine tert-butyl ester; γ-secretase inhibitor X (GSI X), {1S-benzyl-4R-[1-(1S-carbamoyl-2-phenylethylcarbamoyl)-1S-3-methylbutylcarbamoyl]-2R-hydroxy-5-phenylpentyl}carbamic acid tert-butyl ester; γ-secretase inhibitor XI (GSI XI), 7-amino-4-chloro-3-methoxyisocoumarin; γ-secretase inhibitor XII (GSIXII), Z-Ile-Leu-CHO; γ-secretase inhibitor XIII (GSI XIII), Z-Tyr-Ile-Leu-CHO; γ-secretase inhibitor XIV (GSI XIV), Z-Cys(t-Bu)-Ile-Leu-CHO; γ-secretase inhibitor XVI (GSI XVI), N-[N-3,5-difluorophenylacetyl]-L-alanyl-S-phenylglycine methyl ester; γ-secretase inhibitor XVII (GSI XVII); γ-secretase inhibitor XIX (GSI XIX), benzo[e][l,4]diazepin-3-yl)-butyramide; γ-secretase inhibitor XX (GSIXX), (S,S)-2-[2-(3,5-difluorophenyl)acetamido]-N-(5-methyl-6-oxo-6,7-dihydro-5H-dibenzo[b,d]azepin-7-yl)propanamide; γ-secretase inhibitor XXI (GSI XXI), N-[N-3,5-difluorophenylacetyl]-L-alanyl-S-phenylglycine methyl ester; XXI), (S,S)-2-[2-(3,5-difluorophenyl)-acetylamino]-N-(l-methyl-2-oxo-5-phenyl-2-,3-dihydro-lH-benzo[e][l,4]diazepin-3-yl)-propionamide; γ40 secretase inhibitor I, N-trans-3,5-dimethoxycinnamoyl-Ile-leucine;γ40 secretase inhibitor II, N-tert-butyloxycarbonyl-Gly-Val-valine; Isovaleryl-V V-Sta-A-Sta-OCH3; MK-0752 (Merck); MRK-003 (Merck); Semaxit / LY450139 (Eli Lilly) Lilly); RO4929097; PF-03084014; BMS-708163; MPC-7869 (γ-secretase modifier), YO-01027 (dibenzazepine); LY411575 (Eli Lilly); L-685458 (Sigma-Aldrich); BMS-289948 (4-chloro-N-(2,5-difluorophenyl)-N-((lR)-{4-fluoro-2-[3-(lH-imidazol-l-yl)propyl]phenyl}ethyl)benzenesulfonamide hydrochloride); or BMS-299897 (4-[2-((lR)-l-{[(4-chlorophenyl)sulfonyl]-2,5-difluoroaniline}ethyl)-5-fluorophenylbutyric acid) (Bristol Myers Squibb Company Squibb);

[0687] 9. Treatment Indications

[0688] The BCMA binding molecules disclosed herein can be used to treat any disease associated with BCMA expression. In one aspect, the present disclosure provides a method for treating cancer in a subject. The method includes administering a BCMA binding molecule to a subject, thereby treating the subject's cancer. Examples of cancers that can be treated by BCMA targeting agents are cancers associated with BCMA expression, such as multiple myeloma (also known as MM) (see Claudio et al., 2002, Blood [blood]. 100 (6): 2175-86; and Novak et al., 2004, Blood [blood]. 103 (2): 689-94). Multiple myeloma, also known as plasma cell myeloma or Kahler's disease, is a cancer characterized by the accumulation of abnormal or malignant plasma B cells in the bone marrow. Typically, cancer cells invade adjacent bones, destroy bone structure and cause bone pain and fractures. Most cases of myeloma are also characterized by the production of paraproteins (also known as M proteins or myeloma proteins), which are abnormal immunoglobulins produced by excessive clonal proliferation of malignant plasma cells. According to the diagnostic criteria of the International Myeloma Working Group (IMWG), multiple myeloma is diagnosed when the serum paraprotein level exceeds 30 g / L (see Kyle et al., 2009, Leukemia. [Leukemia] 23: 3-9). Other symptoms or signs of multiple myeloma include renal impairment or renal failure, bone lesions, anemia, hypercalcemia, and neurological symptoms.

[0689] 10. Patient groups

[0690] BCMA binding molecules can be used to treat subjects in need. The subject may be diagnosed with cancer, such as a blood cancer, such as multiple myeloma. In some embodiments, the subject may have been previously treated with one or more therapeutic agents. In some embodiments, treatment may fail.

[0691] In some embodiments, the subject has previously received one or more prior treatments for their disease. In some embodiments, the subject has previously received a prior treatment for their disease. In some embodiments, the subject has previously received a prior treatment for their disease. In some embodiments, the subject has previously received a prior treatment for their disease. In some embodiments, the subject has previously received a prior treatment for their disease.

[0692] In some embodiments, the subject has previously received an IMiD, a proteasome inhibitor, an anti-CD38 antibody, or any combination thereof. In some embodiments, the subject has previously received an IMiD. In some embodiments, the subject has previously received a proteasome inhibitor. In some embodiments, the subject has previously received an anti-CD38 antibody.

[0693] 10.1. Inclusion criteria

[0694] Subjects who can be treated with the BCMA-binding molecule may include subjects who have signed an informed consent form prior to treatment with the BCMA-binding molecule.

[0695] Subjects that can be treated with the BCMA-binding molecules may include subjects who are male or female subjects 18 years of age or older.

[0696] Subjects that can be treated with the BCMA-binding molecule may include subjects with an Eastern Cooperative Oncology Group (ECOG) performance status of less than or equal to two (2). The ECOG performance status can be determined at any time prior to treatment with the BCMA-binding molecule.

[0697] The subject that can be treated with the BCMA binding molecules can include a subject with a confirmed cancer diagnosis. For example, the subject that can be treated with the BCMA binding molecules can be a subject with a confirmed multiple myeloma diagnosis. The subject can also receive two or more standard of care (SoC) schemes. The SoC scheme can include IMiD (such as lenalidomide or pomalidomide), proteasome inhibitors (such as bortezomib, carfilzomib) and / or anti-CD38 agents (such as daratumumab. The subject can also have relapse and / or refractory or intolerance to each scheme. Even after receiving previous treatment, the subject may also have written evidence of disease progression (IMWG standards). The subject can also previously receive prior autologous bone marrow transplantation, BCMACAR-T or BCMA-ADC.

[0698] Subjects that can be treated with the BCMA-binding molecules may include subjects with measurable disease, defined by serum M-protein levels greater than or equal to 1.0 g / dL. Subjects that can be treated with the BCMA-binding molecules may include subjects with measurable disease, defined by urine M-protein levels greater than or equal to 200 mg / 24 hours. Subjects that can be treated with the BCMA-binding molecules may include subjects with measurable disease, defined by serum free light chains (sFLC) greater than 100 mg / L of affected FLC.

[0699] Subjects that can be treated with the BCMA binding molecules may include subjects who are willing to undergo continuous bone marrow aspiration and / or biopsy. Continuous bone marrow aspiration and / or biopsy can be performed at any time before treatment with the BCMA binding molecules. Continuous bone marrow aspiration and / or biopsy can be performed at any time after treatment with the BCMA binding molecules. Continuous bone marrow aspiration and / or biopsy can be performed to assess disease status and biomarkers / pharmacodynamics.

[0700] 10.2. Exclusion criteria

[0701] In some embodiments, the subject that can be treated with the BCMA-binding molecule may not have or have one or more of the following exclusion criteria disclosed in this section 10.2. For example, in some embodiments, if the subject has or has any of the following exclusion criteria disclosed in this section 10.2, the BCMA-binding molecule should not be used to treat it. As another example, the subject that can be treated with the BCMA-binding molecule may not have or have two or more of the following exclusion criteria disclosed in this section 10.2. As another example, the subject that can be treated with the BCMA-binding molecule may not have or have three or more of the following exclusion criteria disclosed in this section 10.2. As another example, the subject that can be treated with the BCMA-binding molecule may not have or have four or more of the following exclusion criteria disclosed in this section 10.2. As another example, the subject that can be treated with the BCMA-binding molecule may not have or have five or more of the following exclusion criteria disclosed in this section 10.2.

[0702] Subjects that may be treated with the BCMA-binding molecules may include subjects that may not have undergone prior radiation therapy. In other embodiments, the subject may have received prior radiation therapy. In some embodiments, radiation therapy is not performed within one month of the start of treatment. In some embodiments, radiation therapy is not performed within three weeks of the start of treatment. In some embodiments, radiation therapy is not performed within two weeks of the start of treatment. In some embodiments, radiation therapy is not performed within one week of the start of treatment.

[0703] Some exceptions to prior radiation therapy may be used, for example, if the radiation therapy is localized. For example, localized radiation therapy may be used for bone lesions, such as osteolytic lesions. Or in some cases, localized radiation therapy may be used for plasmacytomas. In these cases, the subject may be eligible for treatment with the BCMA-binding molecule.

[0704] Subjects that may be treated with the BCMA-binding molecules may include subjects who may not have had major surgery recently. In some embodiments, the most recent major surgery is not completed within six months of the start of treatment. In some embodiments, the most recent major surgery is not completed within five months of the start of treatment. In some embodiments, the most recent major surgery is not completed within four months of the start of treatment. In some embodiments, the most recent major surgery is not completed within three months of the start of treatment. In some embodiments, the most recent major surgery is not completed within two months of the start of treatment. In some embodiments, the most recent major surgery is not completed within one month of the start of treatment. In some embodiments, the most recent major surgery is not completed within three weeks of the start of treatment. In some embodiments, the most recent major surgery is not completed within two weeks of the start of treatment. In some embodiments, the most recent major surgery is not completed within one week of the start of treatment.

[0705] Subjects that can be treated with the BCMA-binding molecules may include subjects that may not use steroid therapy. In some embodiments, the steroid can be prednisone, dexamethasone, cortisol, its equivalent or any other corticosteroid for human use. In some embodiments, steroid therapy should not be chronic steroid therapy. For example, the use of 10 mg or more of prednisone or equivalent per day can be considered as chronic steroid therapy.

[0706] Some exceptions may be made if the steroid is administered topically, by inhalation, nasally, or ophthalmically. In these cases, the subject may be eligible for treatment with the BCMA-binding molecule.

[0707] Subjects that may be treated with the BCMA-binding molecules may include subjects who may not be using any immunosuppressive therapy / drug. In some embodiments, the immunosuppressive therapy / drug may not be administered within one month of treatment with the BCMA-binding molecules. In some embodiments, the immunosuppressive therapy / drug may not be administered within four weeks of treatment with the BCMA-binding molecules. In some embodiments, the immunosuppressive therapy / drug may not be administered within three weeks of treatment with the BCMA-binding molecules. In some embodiments, the immunosuppressive therapy / drug may not be administered within two weeks of treatment with the BCMA-binding molecules. In some embodiments, the immunosuppressive therapy / drug may not be administered within one week of treatment with the BCMA-binding molecules. In some embodiments, the immunosuppressive drug treatment is not systemic treatment.

[0708] With respect to steroid therapy and / or immunosuppressive therapy, these considerations are independent of potential pre-, co-, or post-treatment with an immunosuppressive agent to prevent / mitigate any side effects (e.g., CRS) associated with treatment with a BCMA-binding molecule. In other words, a person who receives immunosuppressive therapy before / co-treated / after as part of a treatment regimen comprising a BCMA-binding molecule can still be treated with the BCMA-binding molecule.

[0709] Subjects that can be treated with the BCMA-binding molecules may include subjects who may not have used any BCMAxCD3 bispecific antibody therapy in the past.

[0710] Subjects that can be treated with the BCMA-binding molecules may include subjects who may not have or have had a history of hypersensitivity reactions to any of the ingredients comprising the BCMA-binding molecules. For example, the subject may not have or have had a hypersensitivity reaction to any excipient in the formulation. In some embodiments, the subject may not have or have had a hypersensitivity reaction to other monoclonal antibodies. In some embodiments, the hypersensitivity reaction is a severe hypersensitivity reaction.

[0711] Subjects that may be treated with the BCMA-binding molecules may include subjects who may not have experienced toxicity from any previously treated BCMA-targeted agents.

[0712] Subjects that can be treated with the BCMA-binding molecule may include subjects that do not suffer from any malignant disease other than the disease being treated with the BCMA-binding molecule. In other words, subjects may include subjects that do not suffer from two or more malignant diseases, one of which is not treated by the BCMA-binding molecule.

[0713] Some exceptions may be made if a malignancy has been previously treated and a complete response / remission of the malignancy has been observed. In other words, if the previous treatment for the malignancy was curative. In some embodiments, the malignancy has not recurred in the past five years. In some embodiments, the malignancy has not recurred in the past four years. In some embodiments, the malignancy has not recurred in the past three years. In some embodiments, the malignancy has not recurred in the past two years. In some embodiments, the malignancy has not recurred in the past year. In some embodiments, the malignancy has not recurred in the past six months. Other exceptions may include subjects with basal cell skin cancer and squamous cell skin cancer that have been completely excised. Other exceptions may include completely excised carcinoma in situ of any type. In these cases, the subject may receive treatment with the BCMA-binding molecule.

[0714] Subjects that can be treated with the BCMA-binding molecules may include subjects who do not have an active autoimmune disease. Subjects that can be treated with the BCMA-binding molecules may include subjects who are not known to have an autoimmune disease. Subjects that can be treated with the BCMA-binding molecules may include subjects who are not suspected of having an autoimmune disease.

[0715] Some exceptions for autoimmune diseases can be made for subjects with vitiligo, hypothyroidism, or psoriasis. If a subject has hypothyroidism, the subject may have residual hypothyroidism. In some embodiments, if a subject has residual hypothyroidism, the subject who can be treated with the BCMA-binding molecule only requires hormone replacement. If a subject has psoriasis, the subject who can be treated with the BCMA-binding molecule does not require systemic treatment. In some embodiments, if a subject has psoriasis, the condition is not expected to recur. In these cases, the subject can receive treatment with the BCMA-binding molecule.

[0716] The subjects that can be treated with the BCMA-binding molecules may include subjects that have not been treated with prohibited drugs. In some embodiments, the subject is not treated with prohibited drugs, and the prohibited drugs cannot be discontinued for at least three months before the start of treatment. In some embodiments, the subject is not treated with prohibited drugs, and the prohibited drugs cannot be discontinued for at least two months before the start of treatment. In some embodiments, the subject is not treated with prohibited drugs, and the prohibited drugs cannot be discontinued for at least one month before the start of treatment. In some embodiments, the subject is not treated with prohibited drugs, and the prohibited drugs cannot be discontinued for at least four weeks before the start of treatment. In some embodiments, the subject is not treated with prohibited drugs, and the prohibited drugs cannot be discontinued for at least three weeks before the start of treatment. In some embodiments, the subject is not treated with prohibited drugs, and the prohibited drugs cannot be discontinued for at least two weeks before the start of treatment. In some embodiments, the subject is not treated with prohibited drugs, and the prohibited drugs cannot be discontinued for at least one week before the start of treatment. In some embodiments, the subject is not treated with prohibited drugs, and the prohibited drugs cannot be discontinued for at least six days before the start of treatment. In some embodiments, the subject is not treated with prohibited drugs, and the prohibited drugs cannot be discontinued for at least five days before the start of treatment. In some embodiments, the subject is not treated with a prohibited drug, and the prohibited drug cannot be discontinued for at least four days before the start of treatment. In some embodiments, the subject is not treated with a prohibited drug, and the prohibited drug cannot be discontinued for at least three days before the start of treatment. In some embodiments, the subject is not treated with a prohibited drug, and the prohibited drug cannot be discontinued for at least two days before the start of treatment. In some embodiments, the subject is not treated with a prohibited drug, and the prohibited drug cannot be discontinued for at least one day before the start of treatment.

[0717] Subjects that can be treated with the BCMA-binding molecules can include subjects who do not have grade 2 or greater neuropathy.

[0718] Subjects who can be treated with the BCMA-binding molecules can include subjects who do not have residual toxic effects greater than or equal to Grade 1 from any prior therapy.

[0719] Subjects that can be treated with the BCMA-binding molecules can include subjects who do not have plasma cell leukemia and other plasmacytoid disorders other than multiple myeloma.

[0720] Subjects that can be treated with the BCMA-binding molecules may include subjects who do not have clinical laboratory results with an absolute neutrophil count (ANC) greater than 1,000 / mm3 without growth factor support. The ANC count can be measured 1 month before the start of treatment. In some embodiments, the ANC count can be measured 4 weeks before the start of treatment. In some embodiments, the ANC count can be measured 3 weeks before the start of treatment. In some embodiments, the ANC count can be measured 2 weeks before the start of treatment. In some embodiments, the ANC count can be measured 1 week before the start of treatment. In some embodiments, the ANC count can be measured 6 days before the start of treatment. In some embodiments, the ANC count can be measured 5 days before the start of treatment. In some embodiments, the ANC count can be measured 4 days before the start of treatment. In some embodiments, the ANC count can be measured 3 days before the start of treatment. In some embodiments, the ANC count can be measured 2 days before the start of treatment. In some embodiments, the ANC count can be measured 1 day before the start of treatment.

[0721] Subjects that can be treated with the BCMA-binding molecule may include subjects who do not have clinical laboratory results with a platelet count of less than 75,000 mm3 without transfusion support. The platelet count can be measured 1 month before the start of treatment. In some embodiments, the platelet count can be measured 4 weeks before the start of treatment. In some embodiments, the platelet count can be measured 3 weeks before the start of treatment. In some embodiments, the platelet count can be measured 2 weeks before the start of treatment. In some embodiments, the platelet count can be measured 1 week before the start of treatment. In some embodiments, the platelet count can be measured 6 days before the start of treatment. In some embodiments, the platelet count can be measured 5 days before the start of treatment. In some embodiments, the platelet count can be measured 4 days before the start of treatment. In some embodiments, the platelet count can be measured 3 days before the start of treatment. In some embodiments, the platelet count can be measured 2 days before the start of treatment. In some embodiments, the platelet count can be measured 1 day before the start of treatment.

[0722] The subject that can be treated with the BCMA-binding molecule may include a subject that does not have a clinical laboratory result of a bilirubin level higher than 1.5 times the upper limit of the normal range (ULN). In some embodiments, the bilirubin level may be higher than 1.1 times of ULN. In some embodiments, the bilirubin level may be higher than 1.2 times of ULN. In some embodiments, the bilirubin level may be higher than 1.3 times of ULN. In some embodiments, the bilirubin level may be higher than 1.4 times of ULN. In some embodiments, the bilirubin level may be higher than 1.6 times of ULN. In some embodiments, the bilirubin level may be higher than 1.7 times of ULN. In some embodiments, the bilirubin level may be higher than 1.8 times of ULN. In some embodiments, the bilirubin level may be higher than 1.9 times of ULN. In some embodiments, the bilirubin level may be higher than 2.0 times of ULN.

[0723] The subject that can be treated with the BCMA-binding molecule may include a subject that does not have a clinical laboratory result of an aspartate aminotransferase (AST) level that is higher than 3 times the upper limit of the normal range (ULN). In some embodiments, the AST level may be higher than 1.5 times of the ULN. In some embodiments, the AST level may be higher than 2.0 times of the ULN. In some embodiments, the AST level may be higher than 2.5 times of the ULN. In some embodiments, the AST level may be higher than 3.5 times of the ULN. In some embodiments, the AST level may be higher than 4.0 times of the ULN. In some embodiments, the AST level may be higher than 4.5 times of the ULN. In some embodiments, the AST level may be higher than 5.0 times of the ULN.

[0724] The subject that can be treated with the BCMA-binding molecule may include a subject that does not have a clinical laboratory result of an alanine aminotransferase (ALT) level that is higher than 3 times the upper limit of the normal range (ULN). In some embodiments, the ALT level may be higher than 1.5 times of the ULN. In some embodiments, the ALT level may be higher than 2.0 times of the ULN. In some embodiments, the ALT level may be higher than 2.5 times of the ULN. In some embodiments, the ALT level may be higher than 3.5 times of the ULN. In some embodiments, the AST level may be higher than 4.0 times of the ULN. In some embodiments, the ALT level may be higher than 4.5 times of the ULN. In some embodiments, the ALT level may be higher than 5.0 times of the ULN.

[0725] Subjects that can be treated with the BCMA-binding molecules may include subjects who do not have a clinical laboratory result with a calculated creatinine clearance of less than 30 ml / min. In some embodiments, the calculated creatinine clearance is less than 10 ml / min. In some embodiments, the calculated creatinine clearance is less than 20 ml / min. In some embodiments, the calculated creatinine clearance is less than 40 ml / min. In some embodiments, the calculated creatinine clearance is less than 50 ml / min. The calculated creatinine clearance can be measured by any known method. For example, the Cockcroft-Gault equation can be used to calculate creatinine clearance.

[0726] The subject that can be treated with the BCMA-binding molecule may include a subject that does not suffer from impaired cardiac function. The subject that can be treated with the BCMA-binding molecule does not suffer from clinically significant heart disease. For example, the subject does not suffer from clinically significant and / or uncontrolled heart disease, such as congestive heart failure (e.g., NYHA grade ≥2) that needs to be treated, uncontrolled hypertension, or clinically significant arrhythmias. In some embodiments, the subject does not have QTcF>470msec when screening ECG or congenital long QT syndrome. In some embodiments, less than 3 months before treatment, the subject does not suffer from acute myocardial infarction or unstable angina.

[0727] Subjects that can be treated with the BCMA-binding molecules may include subjects without active infections. In some embodiments, the subject does not have an active infection that requires systemic treatment. In some embodiments, the subject does not have any serious infection within one month prior to treatment. In some embodiments, the subject does not have any serious infection within four weeks prior to treatment. In some embodiments, the subject does not have any serious infection within three weeks prior to treatment. In some embodiments, the subject does not have any serious infection within two weeks prior to treatment. In some embodiments, the subject does not have any serious infection within one week prior to treatment.

[0728] Subjects that can be treated with the BCMA-binding molecules can include subjects who do not have POEMS syndrome (plasma cell dyscrasia with polyneuropathy, organomegaly, endocrinopathy, monoclonal protein, skin lesions).

[0729] Subjects that can be treated with the BCMA-binding molecules can include subjects without any prior allogeneic SCT.

[0730] Subjects that can be treated with the BCMA-binding molecules can include subjects without human immunodeficiency virus (HIV infection).

[0731] Subjects that can be treated with the BCMA-binding molecules may include subjects without active hepatitis B (HBV) or hepatitis C (HCV) infection. Some exceptions may be made to the HBV / HCV requirements if the disease is controlled under antiviral therapy. In some cases, for example, if HBV or HCV is clinically indicated or if the patient has a history of HBV or HCV infection, HBV / HCV should be tested.

[0732] Subjects that can be treated with the BCMA-binding molecules may include subjects who will not use any live vaccines for infectious diseases during treatment. In some embodiments, the subject will not use any live vaccines within 2 weeks of the start of treatment. In some embodiments, the subject will not use any live vaccines within 3 weeks of the start of treatment. In some embodiments, the subject will not use any live vaccines within 4 weeks of the start of treatment. In some embodiments, the subject will not use any live vaccines within 1 month of the start of treatment. In some embodiments, the subject will not use any live vaccines within 2 months of the start of treatment. In some embodiments, the subject will not use any live vaccines within 3 months of the start of treatment.

[0733] Subjects that can be treated with BCMA-binding molecules may include subjects who have not been treated with cytotoxic or small molecule targeted anti-tumor drugs or any experimental therapy before treatment. In some embodiments, within 1 month before starting treatment with the BCMA-binding molecule, the subject has not been treated with cytotoxic or small molecule targeted anti-tumor drugs or any experimental therapy. In some embodiments, within 4 weeks before starting treatment with the BCMA-binding molecule, the subject has not been treated with cytotoxic or small molecule targeted anti-tumor drugs or any experimental therapy. In some embodiments, within 3 weeks before starting treatment with the BCMA-binding molecule, the subject has not been treated with cytotoxic or small molecule targeted anti-tumor drugs or any experimental therapy. In some embodiments, within 2 weeks before starting treatment with the BCMA-binding molecule, the subject has not been treated with cytotoxic or small molecule targeted anti-tumor drugs or any experimental therapy. In some embodiments, within 1 week before starting treatment with the BCMA-binding molecule, the subject has not been treated with cytotoxic or small molecule targeted anti-tumor drugs or any experimental therapy. In some embodiments, the subject has not been treated with cytotoxic or small molecule targeted anti-tumor drugs or any experimental therapy within 10 half-lives before starting treatment with the BCMA-binding molecule. In some embodiments, the subject has not been treated with cytotoxic or small molecule targeted anti-tumor drugs or any experimental therapy within 7 half-lives before starting treatment with the BCMA-binding molecule. In some embodiments, the subject has not been treated with cytotoxic or small molecule targeted anti-tumor drugs or any experimental therapy within 5 half-lives before starting treatment with the BCMA-binding molecule. In some embodiments, the subject has not been treated with cytotoxic or small molecule targeted anti-tumor drugs or any experimental therapy within 4 half-lives before starting treatment with the BCMA-binding molecule. In some embodiments, the subject has not been treated with cytotoxic or small molecule targeted anti-tumor drugs or any experimental therapy within 3 half-lives before starting treatment with the BCMA-binding molecule. In some embodiments, the subject has not been treated with cytotoxic or small molecule targeted anti-tumor drugs or any experimental therapy within 2 half-lives before starting treatment with the BCMA-binding molecule.

[0734] Subjects that can be treated with the BCMA-binding molecules may include subjects who have not induced hematopoietic colony stimulating growth factors (e.g., G-CSF, M-CSF), thrombopoietin mimetics, or erythrocyte stimulating agents less than or equal to two weeks before the start of treatment. In some cases, the initiation does not occur less than one month before the start of treatment. In some cases, the initiation does not occur less than four weeks before the start of treatment. In some cases, the initiation does not occur less than three weeks before the start of treatment. In some cases, the initiation does not occur less than one week before the start of treatment.

[0735] Subjects may receive the BCMA-binding molecule if they received a thrombopoietin mimetic more than two weeks prior to treatment with the BCMA-binding molecule and the subject is stable on the dose.

[0736] Subjects that can be treated with the BCMA-binding molecules can include subjects that have not yet received GM-CSF.

[0737] Subjects that can be treated with the BCMA-binding molecule may include subjects who have not yet received intravenous IG infusions given to prevent infection. In some embodiments, the IG infusion should end 3 months before starting treatment with the BCMA-binding molecule. In some embodiments, the IG infusion should end 2 months before starting treatment with the BCMA-binding molecule. In some embodiments, the IG infusion should end 1 month before starting treatment with the BCMA-binding molecule. In some embodiments, the IG infusion should end 4 weeks before starting treatment with the BCMA-binding molecule. In some embodiments, the IG infusion should end 3 weeks before starting treatment with the BCMA-binding molecule. In some embodiments, the IG infusion should end 4 weeks before starting treatment with the BCMA-binding molecule. In some embodiments, the IG infusion should end 1 week before starting treatment with the BCMA-binding molecule.

[0738] Subjects that can be treated with the BCMA-binding molecules may include subjects who have active central nervous system (CNS) involvement due to malignancy, or symptomatic CNS metastasis, or the presence of CNS metastasis requiring local CNS directed therapy (e.g., radiotherapy or surgery), or an increase in the dose of corticosteroids within 2 weeks prior to the start of treatment. In some embodiments, the CNS problem should not occur 3 months prior to the start of treatment. In some embodiments, the CNS problem should not occur 2 months prior to the start of treatment. In some embodiments, the CNS problem should not occur 1 month prior to the start of treatment. In some embodiments, the CNS should not occur 4 weeks prior to the start of treatment. In some embodiments, the CNS should not occur 3 weeks prior to the start of treatment. In some embodiments, the CNS should not occur 1 week prior to the start of treatment.

[0739] Subjects that can be treated with the BCMA-binding molecules can include subjects suffering from any serious medical or psychiatric illness that may interfere with treatment with the BCMA-binding molecules.

[0740] Subjects that can be treated with the BCMA-binding molecules can include subjects that are not pregnant or lactating (lactating) women. Pregnancy can be defined as the state of a woman after conception until the termination of pregnancy, as confirmed by a positive hCG laboratory test.

[0741] In some embodiments, subjects who can be treated with the BCMA-binding molecules are not women of childbearing potential unless they use effective contraceptive methods (e.g., two, including a highly effective method) during dosing and for 6 months after the last dose of the study drug. Women of childbearing potential can be defined as all women who are physiologically capable of becoming pregnant. Women can be considered postmenopausal and infertile if they have 12 months of natural (spontaneous) amenorrhea and have an appropriate clinical profile (i.e., age-appropriate, history of vasomotor symptoms) or have undergone surgical bilateral oophorectomy (with or without hysterectomy), total hysterectomy, or tubal ligation at least six weeks before. In the case of a single oophorectomy, it is considered to have no fertility potential only when the female reproductive status has been confirmed by subsequent hormone level assessment.

[0742] Highly effective methods of contraception include, but are not limited to, total abstinence; female sterilization; male sterilization; and use of oral, injectable, or implantable hormonal contraceptive methods or placement of an intrauterine device (IUD) or intrauterine system (IUS) and other forms of hormonal contraception with similar efficacy (failure rate <1%) (e.g., hormonal vaginal ring or transdermal hormonal contraception). Other effective methods of contraception include barrier methods such as condoms or occlusive caps (diaphragms or cervical / vault caps) and spermicides (e.g., foams, gels, films, creams, or vaginal suppositories).

[0743] With regard to abstinence, cyclic abstinence (eg, calendar, ovulation-based, temperature-based, postovulation methods) and withdrawal are not acceptable methods of contraception.

[0744] With regard to female sterilization, examples include, but are not limited to, surgical bilateral oophorectomy, with or without hysterectomy), total hysterectomy, or tubal ligation at least six weeks prior to study treatment. In the case of oophorectomy alone, this is only done if the female's reproductive status has been confirmed by subsequent hormone level assessment.

[0745] For male sterilization, this must have occurred at least 6 months prior to screening. For female subjects, the vasectomized male partner should be the subject's only partner.

[0746] With respect to the use of oral contraceptives, in some embodiments, a woman must be stable on the same pill for a minimum of three months prior to starting treatment with the BCMA-binding molecule.

[0747] 11. Examples

[0748] 11.1. Example 1: Identification of BSBM3

[0749] BCMA is a cell surface receptor expressed on plasma cells and other B cell malignancies, particularly multiple myeloma. For efficient drug development, it is highly desirable to have antibodies that cross-react with both human antigens as well as the corresponding antigens in model non-human primate species (e.g., cynomolgus monkeys) for the purpose of nonclinical pharmacokinetic and toxicology studies.

[0750] To identify antibodies that cross-react with both human and cynomolgus BCMA, an initial phage library containing human antibody fragments was subjected to four rounds of panning against recombinant human and cynomolgus BCMA antigens. Approximately 400 single phage colonies were picked from the fourth round of panning, and nine unique clones were selected for amplification and rescue as phage for phage ELISA. The clones were analyzed for affinity to human and cynomolgus BCMA.

[0751] One of the clones was affinity matured as a yeast surface scFv. After multiple rounds of screening, the affinity-matured anti-BCMA pool was cloned into a heterodimeric bispecific antibody format ( Figure 1 ), which were expressed in HEK 293 cells and tested for their ability to bind BCMA on tumor cells and to activate T cells in a target-dependent manner using the Jurkat NFAT luciferase (JNL) reporter gene assay.

[0752] From these assays, a bispecific binding molecule referred to herein as BSBM3 was identified. The sequence of BSBM3 is shown in Table 1 below:

[0753]

[0754]

[0755]

[0756]

[0757] The activity of BSBM3 was compared with that of ch2B4_C29, a BCMA-CD3 bispecific antibody under development for the treatment of multiple myeloma (see, WO 2016 / 0166629). Preliminary data from bivalent BSBM3 and h2B4_C29 from KMS11 and PBMC / T cell co-culture studies showed that bivalent BSBM3 mediated lower levels of cytokine induction compared to h2B4_C29 (data not shown), indicating that patients treated with BSBM3 may have a reduced risk of cytokine release syndrome compared to patients treated with h2B4_C29. Preliminary data also showed that in the presence of KMS11 cells, T cells activated by h2B4_C29 mediated more TCR downregulation than T cells activated by bivalent BSBM3 (data not shown), indicating that BSBM3 can exhibit more persistent anticancer activity than h2B4_C29. Furthermore, in the KMS11 xenograft model, some initial data suggest that BSBM3 (as well as h2B4_C29) has superior anti-tumor activity compared to BCMA-CD3 bispecific molecules from EngMab and Janssen.

[0758] 11.2. Example 2: Characteristics of BSBM3

[0759] BSBM3 is produced in Chinese hamster ovary (CHO) cells and belongs to the IgG1 isotype subclass. Figure 1 As shown, BSBM3 has a Fab domain targeting BCMA, a single-chain Fv (scFv) domain targeting CD3, and the Fc domain confers IgG-like in vivo persistence due to unmodified FcRn (neonatal Fc receptor) affinity. The Fc domain of BSBM3 contains substituents that eliminate binding to human Fcγ receptors and reduce the risk of non-selective T cell activation via FcR (Fc receptor)-mediated cross-linking. Table 2 summarizes its affinity for BCMA and CD3. The simultaneous binding of multiple molecules of BSBM3 to BCMA on multiple myeloma (MM) cells and the CD3 subunit of the T cell receptor (TCR) complex on T cells leads to TCR cross-linking and the formation of cytolytic immune synapses, which leads to the activation of T cells and the specific lysis of MM cells.

[0760]

[0761] Data are shown as mean and SD from three biological replicates.

[0762] 11.3. Example 3: Nonclinical Pharmacology (In Vitro)

[0763] The activity of BSBM3 was characterized in an in vitro co-culture system with BCMA+ myeloma cell line KMS11 and healthy donor T cells. At concentrations ≥1nM, BSBM3 induced T cell proliferation and cytokine secretion (Figure 2). Consistently, BSBM3 mediated effective redirected T cell cytotoxicity (RTCC) to KMS11 in a concentration-dependent manner (Figure 2). In contrast, the non-targeted control antibody NT-CD3 (with the same anti-CD3 scFv, but non-targeted Fab instead of anti-BCMA Fab) did not induce T cell proliferation or significant killing of KMS11 cells, indicating that specific binding to BCMA on tumor cells is required for T cell activation and cytotoxicity. These data show that in the presence of BCMA+ cells, BSBM3 can effectively and specifically activate T cells, resulting in specific killing of target cells.

[0764] To determine the in vitro assay that provided the most sensitive readout for BSBM3 activity, EC30 values ​​were calculated from different assays, each of which was performed with nine biological replicates. (T cells from three healthy donors were tested, each replicated in three independent experiments; Figure 3 ). The redirected T cell cytotoxicity (RTCC) assay detects specific lysis of MM cells and shows the most sensitive and reproducible EC30 values. Therefore, the minimum expected biological effect level (MABEL) that informs the starting dose was calculated based on the EC30 values ​​from the RTCC assay.

[0765] BCMA has been shown to be protease cleaved within its transmembrane domain by γ-secretase, resulting in the shedding of its extracellular domain as a soluble factor (hereinafter referred to as soluble BCMA), which is used as a bait to neutralize its ligand APRIL (Laurent 2015). It is reported that the average serum level of soluble BCMA is 39 ng / mL in healthy subjects, 89 ng / mL in subjects with smoldering myeloma, and 506 ng / mL in newly diagnosed MM subjects (Ghermezi et al., 2017, Haematologica. [Hematology] 102 (4): 785-795). Soluble BCMA in the blood and bone marrow of subjects has the potential to bind to BSBM3 and interfere with the activity of BSBM3. As expected, in the presence of 30, 100 or 300 ng / mL of soluble BCMA, the EC30 of BSBM3 increased by 6 times, 15 times, and 41 times, respectively ( Figure 4). Because 93% of subjects with active and untreated MM have shed BCMA above 107.6 ng / mL in their serum (Ghermezi et al., 2017, Haematologica [Hematology] 102(4):785-795), the RTCC assay containing 100 ng / mL of soluble BCMA may better represent the activity of BSBM3 in the subjects. Therefore, when calculating the starting dose using the MABEL method, an EC30 of 0.753 ng / mL (at 100 ng / mL soluble BCMA) should be considered. This is consistent with the approach described by Saber et al., 2017, Regul Toxicol Pharmacol [Regulatory Toxicology and Pharmacology]; 90:144-152 for CD3-directed bispecific antibodies.

[0766] 11.4. Example 4: Nonclinical Pharmacology (In Vivo)

[0767] Using the KMS11 xenograft model, the in vivo activity of BSBM3 was evaluated in immunocompromised NSG mice adoptively transferred with human PBMCs from healthy donors ( Figure 5 ). KMS11 cells were engineered to overexpress luciferase, and tumor burden was then measured by bioluminescence intensity (BLI). In three independent experiments (using PBMCs from two different healthy donors), mice treated with BSBM3 at doses of ≥ 0.3 mg / kg showed strong tumor rejection (data).

[0768] The adoptive transfer model with KMS11 xenografts provides support for the mechanism of action of BSBM3. However, it may over-predict anti-MM activity because adoptively transferred human T cells are extremely active, as indicated by significantly higher expression of activation markers compared to T cells in donor PBMCs after isolation (data show; Ali et al., 2012, PLoS ONE [Public Library of Science Comprehensive]; 7(8): e44219). Therefore, the dose that demonstrated anti-MM activity of BSBM3 in this model cannot be directly transferred to subjects.

[0769] 11.5. Example 5: Nonclinical Pharmacokinetics and Metabolism

[0770] To investigate the pharmacokinetics (PK) of BSBM3 in a non-binding species, NSG mouse PK studies were performed in the presence or absence of human peripheral blood mononuclear cells (PBMCs). The concentration-time plots showed a biexponential decline in serum levels, as expected for monoclonal antibodies in non-binding species. In NSG mice humanized with human PBMCs, exposure as measured by AUClast was lower than in PBMC-naive mice. At the last time point, nonlinear elimination became apparent, indicating the expected target-mediated drug disposition (TMDD).

[0771] BSBM3 binds to two targets (BCMA and CD3) in cynomolgus monkeys, therefore, the toxicokinetic characteristics of BSBM3 were studied in a single-dose non-GLP toxicology study (data not shown) and a 4-week GLP toxicology study (data not shown). From the single-dose study, it was determined that exposure to BSBM3 increased in a dose-proportional manner at the test doses of 0.3, 1, and 3 mg / kg as measured by AUClast. Of the five animals dosed, one animal (0.3 mg / kg dose) was confirmed to have anti-drug antibodies (ADA).

[0772] In a 4-week GLP toxicology study in cynomolgus monkeys, animals received 5 weekly intravenous injections of BSBM3 (at 1, 3, and 10 mg / kg). After intravenous injection, the maximum exposure to BSBM3 was observed at the first time point after administration, 0.667 to 4.17 hr after administration. Exposure to BSBM3, as measured by Cmax and AUC0-tau (tau = 7 days), increased in an approximately dose-proportional manner over the dose range of 1 to 10 mg / kg and was similar in both sexes. At all dose levels, the cumulative amount was approximately 1.5 to 1.9 times (based on AUC0-tau) after 4 weeks of intravenous administration. No significant gender differences were observed. In the main part of the study, ADA was detected on day 28 (1 of 24 treated animals at a dose of 1 mg / kg). In the 6-week recovery portion of the study, ADA was detected on day 57 (1 of 6 treated animals, dosed at 3 mg / kg) and day 71 (2 of 6 treated animals, control group). The data suggest that ADA may not have a significant effect on TK. ADA was not detected in control animals.

[0773] 11.6. Example 6: Nonclinical Toxicology

[0774] The safety of BSBM3 was investigated in in vitro and in vivo studies.In vivo studies were conducted in cynomolgus monkeys, identified as a pharmacologically relevant species for BSBM3.

[0775] Results from safety pharmacology studies indicate that BSBM3 poses a low risk to the central nervous system (CNS), respiratory and cardiovascular systems. In general, results from in vivo studies were consistent with the expected pharmacology associated with BSBM3 as follows: a decrease in peripheral blood, bone marrow and tissue B cells and plasma cells; and an acute increase after dose of selective serum cytokines; and more persistent blood and tissue T-cell activation. In GLP studies, at all BSBM3 dose levels, it was noted that lymphocytes in the gut-associated lymphoid tissue (GALT), lymph nodes and spleen (B cell area) were depleted. During the recovery phase, lymphocyte hyperplasia in these organs was consistent with the regenerative process. In addition, mixed cellular immune-inflammatory lesions were observed in various organs (i.e., gastrointestinal tract (GIT), liver, spleen, heart, kidney, lungs), and were sometimes associated with infectious agents.

[0776] In the GLP study, the highest non-serious toxic dose (HNSTD) was identified as 1 mg / kg.

[0777] 11.7. Example 7: Clinical study

[0778] Based on Figure 6 A clinical trial of the indicated regimen to determine the safety and efficacy of BSBM3 in subjects with multiple myeloma who have received two or more lines of standard of care (SoC) treatment including an IMiD (e.g., lenalidomide or pomalidomide), a proteasome inhibitor (e.g., bortezomib, carfilzomib), and an anti-CD38 agent (e.g., daratumumab), and who are relapsed and / or refractory or intolerant to each regimen.

[0779] The study consisted of a dose-escalation portion followed by an expansion portion.

[0780] This is a FIH, Phase I, multicenter, open-label study to determine the safety and efficacy of BSBM3 (a bispecific antibody that specifically binds to BCMA and CD3 as described throughout this disclosure) in subjects with multiple myeloma who have received two or more lines of standard of care (SoC) treatment including an IMiD (e.g., lenalidomide or pomalidomide), a proteasome inhibitor (e.g., bortezomib, carfilzomib), and an anti-CD38 agent (e.g., daratumumab), and are relapsed and / or refractory or intolerant to each regimen, have documented evidence of disease progression according to the International Myeloma Working Group (IMWG) criteria, and who are ineligible for treatment with other regimens known to provide clinical benefit.

[0781] 11.7.1. Inclusion criteria

[0782] Subjects included in the trial had an established diagnosis of multiple myeloma and had received two or more standard of care (SoC) regimens including an IMiD (e.g., lenalidomide or pomalidomide), a proteasome inhibitor (e.g., bortezomib, carfilzomib), and an anti-CD38 agent (e.g., daratumumab), if available, and were relapsed and / or refractory or intolerant to each regimen, had documented evidence of disease progression (IMWG criteria), and must not have been eligible for treatment with other regimens known to provide clinical benefit, as determined by the investigator. (Subjects who had received prior autologous bone marrow transplantation, BCMA CART, or BCMA-ADC therapy and who otherwise met the inclusion criteria were eligible to participate in the study); at screening, had an Eastern Cooperative Oncology Group (ECOG) performance status ≤ 2; and had measurable disease defined by at least 1 of the following 3 measurements: (i) serum M-protein ≥ 1.0 g / dL; (ii) urine M-protein ≥ 200 mg / 24 hours; or (iii) serum free light chain (sFLC) > 100 mg / L affected FLC.

[0783] 11.7.2. Exclusion criteria

[0784] Subjects who met any of the following criteria were not eligible for inclusion in this study: radiation therapy within 14 days prior to the first dose of study drug, except for local radiation therapy for lytic lesions or plasmacytomas; major surgery within 2 weeks prior to the first dose of study drug; use of systemic chronic steroid therapy (≥10 mg / day of prednisone or equivalent), or any immunosuppressive therapy (topical, inhaled, nasal, or ophthalmic steroids are allowed) within 7 days of the first dose of study treatment; previous use of BCMAxCD3 bispecific therapy; subjects receiving systemic treatment with any immunosuppressive drug (except steroids as described above); a history of serious hypersensitivity reaction to any component of one or more study drugs and other mAbs and / or their excipients; subjects with prior toxicity from BCMA-targeted agents; malignant disease, except for those being treated in this study. (Exceptions to the exclusions include the following: malignancies that have been treated with curative intent and have not recurred within 2 years prior to study treatment; completely excised basal cell skin cancer and squamous cell skin cancer, and completely excised carcinoma in situ of any type); active, known or suspected autoimmune disease, except for subjects with vitiligo, residual hypothyroidism requiring hormone replacement only, psoriasis not requiring systemic therapy, or conditions not expected to recur; subjects currently receiving contraindicated medications that cannot be discontinued for at least one week prior to the start of treatment; subjects with ≥ Grade 2 neuropathy and residual Toxic effects must have resolved to ≤ Grade 1 or baseline; Subjects with plasma cell leukemia and other plasmacytoid disorders (except MM); Any of the following clinical laboratory results: (i) absolute neutrophil count (ANC) <1,000 / mm3 without growth factor support within 7 days before the start of treatment; (ii) platelet count <75,000 mm3 without transfusion support within 7 days before the start of treatment; (iii) bilirubin >1.5 times the upper limit of normal range (ULN); (iv) aspartate aminotransferase (AST) or alanine aminotransferase (ALT) >2.5 times the ULN.5-fold; (v) creatinine clearance calculated by the Cockcroft-Gault equation <30 ml / min; (vi) impaired cardiac function or clinically significant cardiac disease; active infection requiring systemic therapy or other serious infection within 2 weeks before the first dose of study drug; POEMS syndrome (plasma cell dyscrasia with polyneuropathy, organomegaly, endocrinopathies, monoclonal proteins, skin lesions); previous allogeneic SCT at any time before signing the informed consent for the study; human immunodeficiency virus (HIV infection); active hepatitis B (HBV) or hepatitis C (HCV) infection; use of any live vaccine against infectious diseases (e.g., influenza, varicella, pneumococcus) within 4 weeks of the start of study treatment; treatment with cytotoxic antineoplastic drugs or small molecule targeted antineoplastic drugs or any experimental therapy within 14 days or 5 half-lives, which is less than the time before the first dose of study treatment; induced hematopoiesis ≤ 2 weeks before the start of study treatment Colony stimulating growth factors (e.g., G-CSF, M-CSF), thrombopoietin mimetics, or erythrocyte stimulating agents; intravenous IG infusions given for infection prevention must be discontinued ≥ 28 days prior to the start of study treatment; active central nervous system (CNS) involvement due to malignancy, or the presence of symptomatic CNS metastases or CNS metastases requiring local CNS-directed therapy (e.g., radiation therapy or surgery), or dose increases of corticosteroids within 2 weeks prior to the start of study treatment; serious medical or psychiatric illness that may interfere with participation in this clinical study; pregnant or lactating (lactating) women, where pregnancy is defined as the state of a woman after conception until termination of pregnancy, as confirmed by a positive hCG laboratory test; and women of childbearing potential (defined as all women who are physiologically capable of becoming pregnant), unless at the time of informed consent, they are using two effective methods of contraception (including at least one highly effective method of contraception) during dosing and for a period of 6 months after the last dose of study drug. .

[0785] 11.7.3. Pharmaceutical products

[0786] The drug product was formulated as liquid in vial (LIVI) and consisted of 10 mg / mL BSBM3, 20 mM histidine, 240 mM sucrose, PS20 0.04%, pH 5.5 ± 0.3.

[0787] All doses and all dose changes prescribed and administered to subjects during the study were recorded on the Dosing Administration Record, eCRF.

[0788] Table 3 Study drugs

[0789]

[0790] Even after the expansion part under RD begins, exploration of alternative doses and / or dosing schedules of BSBM3 may be evaluated stepwise. Subjects will be assigned to cohorts at all sites in this overall study in an alternating manner if recruited simultaneously.

[0791] 11.7.4. Treatment process

[0792] BSBM3 will be administered weekly (Q1W) initially. Treatment with study drug will continue until the subject experiences unacceptable toxicity, disease progression per IMWG, or treatment is discontinued at the discretion of the investigator or patient. The study design is summarized in Figure 6 In. Alternative dosing schedules (e.g., Q2W, Q3W, TIW) may be implemented during the study if supported by emerging data (including preliminary PK, PD, and efficacy results from this ongoing trial). If clinically significant cytokine release syndrome (CRS) or related symptoms are observed during dose escalation, initial dosing options may be introduced and subsequent dosing schedules modified.

[0793] The design of this Phase I, open-label study was selected to characterize the safety and tolerability of BSBM3 in subjects with relapsed and / or refractory multiple myeloma who have been treated with at least 2 prior regimens and have received an IMiD, a proteasome inhibitor, and an anti-CD38 antibody (if available), and to determine the recommended dose and schedule for further study. If necessary, the dose escalation allowed the establishment of the MTD of BSBM3 and was guided by a Bayesian logistic regression model (BLRM).

[0794] BLRM is a well-established method for estimating MTD in cancer subjects. Adaptive BLRM will be guided by the principles of escalation with overdose control (EWOC) to control the risk of DLT in future subjects in the study. The use of Bayesian response adaptive models for small data sets has been accepted by the EMEA (“Guideline on clinical trials in small populations”, February 1, 2007) and has been endorsed by numerous publications (Babb et al., 1998, Stat Med; 17(10): 1103-20); (Neuenschwander et al., 2008, Stat Med; 27(13): 2420-39); (Neuenschwander et al., 2010, Clin Trials; 7(1): 5-18); (Neuenschwander et al., 2014, In A Bayesian Industry Approach to Phase I Combination Trials in Oncology. In Statistical Methods in Drug Combination Studies. Zhao W and Yang H (eds.), Chapman and Hall / CRC, 2014), and its development and appropriate use is an aspect of the FDA's Critical Path Program.

[0795] At the dose-escalation meeting, decisions on new dose levels are made based on tolerability and safety information in the subjects, including the DLT risk estimate derived from the BLRM, and a review of the available PK, PD, and initial activity information at the time of the decision.

[0796] 11.7.5. Dose escalation

[0797] During dose escalation, subjects with relapsed and / or refractory MM will receive BSB3 until the MTD / RD is reached. During the escalation process, an estimated 21 subjects will be needed to determine the MTD / RD.

[0798] The safety (including dose-DLT relationship) and tolerability of the study treatment will be evaluated, and based on review of these data, one or more regimens and one or more doses for use in the expansion portion will be determined. The RD will also be guided by available information on PK, PD, and preliminary anti-tumor activity. Dose escalation will be guided by an adaptive Bayesian logistic regression model (BLRM) that follows the principles of dose escalation with overdosing control (EWOC).

[0799] Once one or more MTD / RDs are determined in the escalation part, additional subjects will be recruited in the expansion part to further characterize the PK, PD, and safety characteristics of the study drug and to evaluate the initial anti-tumor activity of BSBM3. More than one dose level under the Q1W regimen may be explored as the RD for expansion. In addition, alternative dosing regimens may be explored in the escalation part. One or more RDs of the new regimen may be stated.

[0800] In the expansion part, subjects with relapsed and / or refractory MM will be treated with BSBM3. The expansion part will enroll approximately 20 subjects. Enrollment may be stopped early based on ongoing review of data from the expansion cohort.

[0801] 11.7.5.1. Tentative Dosing

[0802] The dose of BSBM3 was proposed based on a comprehensive evaluation of the following: predicted pharmacokinetics, mechanism of action, in vitro efficacy (to inform the MABEL dosing approach), the impact of circulating BCMA, and in vivo safety from GLP toxicology studies in cynomolgus monkeys. Subjects were given a starting dose of BSBM3 of 3 mcg / kg, administered as an intravenous infusion over 2 hours.

[0803] The starting dose of BSBM3 and the dose levels that can be evaluated during this trial are described in Table 4. This starting dose is supported by the EC50 value (approximately 0.07 μg / mL) from the RTCC assay (without added recombinant soluble BCMA), which is believed to represent the most clinically relevant measure of pharmacological activity and is the most sensitive and reproducible assay readout for BSBM3 in vitro (data not shown).

[0804] The actual dose level will be determined based on available toxicity, pharmacokinetic, and pharmacodynamic data and will be guided by the BLRM. Dose escalation will continue until one or more MTDs or RDs are determined.

[0805] Table 4 Tentative dose levels

[0806]

[0807]

[0808] Alternatively, if during dose escalation, 2 patients experience an event of Grade ≥ 3 infusion-related reaction (IRR) or cytokine release syndrome (CRS) that does not resolve to Grade ≤ 1 or baseline within 48 hours, the initial dose will be used.

[0809] The initial dose will be selected at a dose level determined to be safe (the dose should be at least one dose level lower than the maximum dose tested in the previous group and meet the EWOC criteria). In addition, as an additional safety measure, one-third of the initial dose will be given on day 1 and two-thirds of the initial dose will be given on day 2. Once the initial dose level is determined, the dose levels that can be evaluated in subsequent groups relative to the initial dose can be determined and listed in Table 5. For example, if the initial dose is determined to be 100mcg / kg (i.e., dose level X in Table 5), the dose on day 1 will be 33.33mcg / kg and the dose on day 2 will be 66.66mcg / kg. The third infusion and subsequent infusions (on days 8, 15 and 22) will be 200mcg / kg (i.e., dose level X+1, where X+1 is the next tentative dose level after X listed in Table 3). The actual dose level will be determined based on available toxicity, pharmacokinetic and pharmacodynamic data. A separate BHLRM will be constructed to guide dose escalation under EWOC criteria. Dose escalation will continue until one or more MTDs or RDs are determined.

[0810] Table 5 Tentative dose levels for the first dose

[0811]

[0812]

[0813] The initial dose level may be changed, if necessary, based on evolving trial safety and tolerability results.

[0814] 11.7.5.2. Criteria for Dose Escalation and Determination of MTD / RD

[0815] Dose escalation is performed to establish one or more doses of BSBM3 to be used in the expansion portion. Specifically, one or more doses are considered to have the most appropriate benefit-risk, taking into account the maximum tolerated dose (MTD), as assessed by review of safety, tolerability, PK, any available efficacy, and PD.

[0816] The MTD is the highest dose assessed to have a risk of less than 25% of dose-limiting toxicity (DLT) in more than 33% of treated subjects during the DLT evaluation period. The dose or doses selected for the expansion portion may be any dose equal to or less than the MTD and may be declared without determining the MTD.

[0817] Each dose escalation cohort will start with 1 to 6 newly treated subjects. They must have adequate exposure and follow-up to be considered evaluable for dose escalation decisions.

[0818] If any subject experiences a DLT during the DLT assessment period, the minimum cohort size will increase to three.

[0819] If one or more subjects discontinue and do not meet the evaluability criteria, alternative strategies can be used to recruit additional subjects into the same cohort to support the benefit-risk assessment.

[0820] The treatment period will start on Day 1 of Cycle 1. For the purpose of scheduling and evaluation, the treatment cycle will consist of 28 days. For each group in which the dose level on Day 1 of Cycle 1 is higher than any dose previously tested and shown to be safe, a staggered approach will be adopted for the first two subjects in the group. After the first subject is administered, the next subject will be administered at least 72 hours after the previous subject is administered. After completing this staggered dosing of the first two subjects, subsequent subjects will be treated without misalignment, however, only 1 patient in the group will receive its first infusion on any given day. When all subjects in the group have completed the DLT assessment period or terminated, a dose escalation decision will be made. The decision will be based on a combination of all relevant data available from all dose levels evaluated in ongoing studies, including safety information, available PK, available PD, and initial efficacy.

[0821] Through the Bayesian logistic regression model (BLRM), any dose escalation decision will not exceed the dose level that meets the EWOC principle. For any dose level, the dose of the next escalation group will not exceed a 100% increase from the previously tested safe dose. Investigators and sponsors can recommend smaller increases in dose after considering all available clinical data.

[0822] To better understand the safety, tolerability, PK, PD, or anti-tumor activity of BSBM3 before or as further escalation is performed, enriched cohorts of 1-6 subjects may be enrolled at any dose level, or at a dose lower than the highest dose previously tested and shown to be safe.

[0823] To reduce the risk of exposing subjects to excessively toxic doses, if 2 subjects experience DLT in a new cohort, the BLRM will be updated with the latest information from all cohorts without waiting for all subjects from the current cohort to complete the assessment period.

[0824] - If the 2 DLTs occur in an escalation cohort, enrollment in that cohort will be stopped and the next cohort will be opened at a lower dose that meets the EWOC criteria.

[0825] - If the 2 DLTs occur in the enriched cohort, additional subjects may be recruited to the open cohort only if the dose still meets the EWOC criteria after re-evaluation of all relevant data. Alternatively, if recruitment to the same dose is not possible, a new cohort of subjects may be recruited to a lower dose that meets the EWOC criteria. In addition, if 2 or more patients experience DLT in a dosing cohort, the next dose escalation level will not exceed 50% of the previous dose level.

[0826] 11.7.6. CRS Management

[0827] Each patient should receive at least 2 doses of tocilizumab before infusion of BSBM3. Hospitals should use additional doses of tocilizumab promptly. Supportive care, tocilizumab, and corticosteroids have been used for effective management of CRS. Prompt responses to tocilizumab have been observed in most subjects.

[0828] Cytokine release syndrome (CRS) was identified based on clinical manifestations (see Table 6). Other causes of fever, hypoxia, and hypotension were evaluated and treated, and subjects were monitored for signs or symptoms of CRS for at least 4 weeks after treatment with BSBM3. If signs or symptoms of CRS occurred at any time, the subject was advised to seek medical attention immediately.

[0829] At the first signs of CRS (see Table 6), patients should be promptly evaluated for hospitalization and treated with supportive care, tocilizumab, and / or corticosteroids as indicated.

[0830] Recommended treatments for the management of CRS are presented below in Tables 7 and 8. The approach to CRS management is a guideline, and investigators may act or modify treatments at their discretion based on the needs of individual subjects.

[0831] Table 6 Clinical signs and symptoms associated with CRS (Lee et al., 2014, Blood 124(2):188-95)

[0832]

[0833] Table 7 CRS Management

[0834]

[0835]

[0836] * If there is no improvement after tocilizumab and steroids, consider other anti-cytokine and anti-T-cell therapies. These therapies may include siltuximab (11 mg / kg intravenously over 1 hour), high-dose steroids (eg, high-dose methylprednisolone or equivalent, with steroid doses determined according to local ICU practice), cyclophosphamide, antithymocyte globulin (ATG), or alemtuzumab.

[0837] Table 8 High-dose vasopressors

[0838]

[0839] *Vasopressin and Septic Shock Trial (VASST) Norepinephrine Equivalent Equation:

[0840] NE dose = [norepinephrine (mcg / min)] + [dopamine (mcg / kg / min) ÷ 2] + [epinephrine (mcg / min)] + [phenylephrine (mcg / min) ÷ 10] (Russell et al., 2008, N Engl J Med; 358(9): 877-87)

[0841] If the subject does not respond to tocilizumab, other anti-cytokine therapies may also be considered based on the availability of other anti-cytokine therapies. If the subject still experiences sustained CRS despite the administration of anti-cytokine directed therapy, anti-T cell therapy (e.g., cyclophosphamide, anti-thymocyte globulin (ATG) or alemtuzumab) may be considered. These therapies are collected in appropriate CRFs.

[0842] Management of CRS is based solely on clinical parameters as described in Table 7. Ferritin, CRP, and serum cytokine levels cannot be used for clinical management decisions. Cases of transient left ventricular dysfunction as assessed by echocardiography (ECHO) have been reported in some subjects with severe CRS (Grade 4). Therefore, consider monitoring cardiac function by ECHO during severe CRS, especially in the setting of prolonged severe hemodynamic instability, delayed response to high-dose vasopressors, and / or severe fluid overload.

[0843] 11.7.7. Primary End Point

[0844] The primary endpoints of this study are listed in Table 9.

[0845] Table 9 Primary endpoints

[0846]

[0847] Dose-limiting toxicity (DLT) was defined as an adverse event or abnormal laboratory value assessed as clinically relevant occurring ≤28 days after the first dose of study treatment.

[0848] 11.7.8. Results

[0849] BSBM3 was found to be safe and well tolerated, and had antitumor activity.

[0850] 11.8. Example 8 Effective Dose Range of AL-102 for BCMA Shedding Inhibition 11.8.1. Overview

[0851] To evaluate the effect of the gamma-secretase inhibitor (GSI) AL-102 on B cell maturation antigen (BCMA) shedding in KMS11 cells in vitro.

[0852] Materials and methods

[0853] 11.8.2.1. GSI treatment of KMS11 cells

[0854] KMS11-Luc cells were plated at 1.5×10 5 Cells / well were cultured in RPMI1640 (Gibco #11875-085) supplemented with 20% FBS (Seradigm #1500-500) and L-glutamine (Thermo Fisher #25030-081) in a final volume of 200 μL, which included 12-point, 5-fold serial dilutions of AL-102. The highest starting concentration of AL-102 was 1 μM. The cells were incubated at 37°C / 5% CO2 for 20 hours. The cells were pelleted, the supernatant was collected for measurement of shed BCMA levels, and the cell pellets were stained to assess BCMA membrane expression levels.

[0855] 11.8.2.2. Measurement of shed BCMA levels by ELISA

[0856] Soluble BCMA levels in the supernatant were determined by ELISA according to the protocol provided by the supplier (R&D Systems #DY193). In brief, recombinant human BCMA-Fc protein was included in the kit and used to generate a standard curve. The collected samples were measured and the sBCMA concentration was extrapolated from the standard curve. The quantitative values ​​as determined by the kit were divided by 5.5 to correct for the molecular weight difference between the BCMA-Fc fusion protein (32,554.6 Da) used as the standard curve in the kit and the mass of the endogenously shed BCMA extracellular domain (5,899.3 Da). The results were analyzed using SoftMax Pro v5.4.1 and plotted in GraphPad Prism. 11.8.2.3. Analysis of BCMA membrane expression by flow cytometry

[0857] The cells were precipitated by centrifugation, and the supernatant was transferred to a fresh plate and frozen at -80 ° C for subsequent sBCMA analysis by ELISA. For membrane BCMA analysis, the cell pellet was resuspended in 100 μL BD staining buffer containing BSA (BD#554657) and stained with anti-BCMA-PE (Biolegend, clone 19F21.25ul / test) and fixable viability dye eFluor506 (ThermoScientific, 1:800 dilution) at 4 ° C for 30 minutes. Samples were analyzed by flow cytometry on BD LSR Fortessa instruments. FlowJo v10 software was used for analysis. Using Quantum Simply Cellular beads (Bangs Laboratories), the anti-BCMA antibody binding capacity (ABC) on KMS11 cells was determined according to the scheme provided by the supplier. ABC is an estimate of the number of receptors per cell. These results were plotted against the concentration of AL-102 in Graphpad Prism software.

[0858] 11.8.3. Results

[0859] AL-102 effectively inhibited BCMA shedding from KMS11 cells in a dose-dependent manner, resulting in increased BCMA expression on the cell surface within the same effective dose range ( Figure 8 ). Untreated KMS11 cells had a BCMA antibody binding capacity (ABC) of approximately 14,000. The average ABC under 1 μM AL-102 treatment was approximately 285,000, and cell surface BCMA expression under AL-102 treatment increased 20-fold.

[0860] 11.9. Example 9: Effect of AL-102 on the efficacy of BSBM3

[0861] 11.9.1. Overview

[0862] To evaluate the ability of AL-102 to enhance BSBM3 activity, a redirected T cell cytotoxicity (RTCC) assay was performed using human T cells and BCMA-expressing multiple myeloma cell lines treated with a dose-range combination of BSBM3 and AL-102 in a 10x8 matrix format.

[0863] Materials and methods

[0864] 11.9.2.1. Isolation of healthy human T cells

[0865] Human T cells were enriched from the peripheral blood of three healthy human donors. First, Ficoll-Paque PLUS density gradient (GE Healthcare #17-1440-02) was used in Leucosep tubes (Greiner #227290) to separate peripheral blood mononuclear cells (PBMC) from donor blood and stored in liquid nitrogen in the form of live frozen aliquots. PBMCs were thawed and all T cells were separated by negative selection according to the manufacturer's recommended protocol (Miltenyi #130-096-535). Unlabeled cell fractions rich in T cells were collected by manual magnetic separation on LS columns (Miltenyi #130-042-401). T cells were prepared in T cell medium (TCM) consisting of RPMI-1640 (Gibco #11875-085), 10% FBS (Seradigm #1500-500), 1% Pen / Strep (Life Technologies #15070063), 1% L-glutamine (Thermo Fisher Scientific #25030-081), 1% non-essential amino acids (NEAA) (Life Technologies #11140-050), sodium pyruvate (NaPy) (Life Technologies #11360-070), HEPES (Life Technologies, catalog #15630080), and 0.1% 2-β-mercaptoethanol (2-BME) (Life Technologies, catalog #21985-023).

[0866] 11.9.2.2. KMS11 Multiple Myeloma Cell Line

[0867] KMS11 multiple myeloma cell line was cultured in RPMI1640 supplemented with 20% FBS (Gibco #11875-085, Seradigm #1500-500).

[0868] 11.9.2.3. Redirected T cell cytotoxicity (RTCC) assay

[0869] The target MM cell line KMS11 was transduced to constitutively express luciferase (KMS11-Luc) and used to measure cell viability / survival rate. The KMS11-Luc cells were precipitated and resuspended in fresh medium just before plating to remove any basal level of shed BCMA that may be present. 7,500 KMS11-Luc target cells in 10 μL TCM were added to the wells of a 384-well plate (Corning #3765). 10 μL of 10 nM concentration of BSBM3 was diluted 5 times, and 10 μL of 1000 nM concentration of AL-102 was diluted 5 times and distributed to the corresponding wells of the assay plate. 15,000 T cells in 10 μL TCM were added to the corresponding wells of the assay plate, and E:T was 2:1. The assay was incubated at 37°C / 5% CO2 for 48 hours, and then the luciferase activity was measured according to the manufacturer's protocol to indicate target cell viability (BrightGlo, Promega #E2650). Plates were read on an Envision microplate reader. Only target cells without T cells or antibodies (KMS11-Luc) were used as controls and represented 100% luciferase activity (100% viability). Data were plotted and analyzed using GraphPadPrism software. EC50 values ​​were calculated using S-type, four-parameter nonlinear regression curve fitting.

[0870] 11.9.3. Results

[0871] RTCC assays were set up with three individual T cell donors cultured with KMS11-Luc cells in the presence of BSBM3 alone or a dose response curve of BSBM3 in combination with AL-102. BSBM3 exhibited a dose-dependent effect on KMS11-Luc cell death with an EC50 value of 1 nM. The combination of BSBM3 with AL-102 increased the killing capacity of BSBM3 ( Fig. 9). AL-102 reduced the EC50 value of BSBM3 from 1 nM to 0.02 nM, indicating that the RTCC activity of BSBM3 was enhanced in the presence of AL-102. This represents a 50-fold increase in BSBM3 potency. AL-102 at 0.32 nM or less had minimal effect on BSBM3 potency, had a moderate effect at 1.6 nM, and showed the greatest enhancement of BSBM3 potency at 8 nM or higher concentrations. These results indicate that the combination of AL-102 and BSBM3 synergistically enhances the RTCC potency of BSBM3.

Claims

1. A method of treating a subject having multiple myeloma, the method comprising administering to the subject one or more therapeutic doses of a bispecific antibody that binds to human BCMA and human CD3 and comprises: (a) a first polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 1; (b) a second polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 2; and (c) a third polypeptide, wherein the amino acid sequence of the third polypeptide comprises the amino acid sequence of SEQ ID NO:

3.

2. The method of claim 1, wherein the subject has measurable disease.

3. The method of claim 2, wherein the subject has a serum M-protein level ≥ 1 g / dL.

4. The method of claim 2 or claim 3, wherein the subject produces a urine M-protein level of ≥ 200 mg / 24 hours.

5. The method according to any one of claims 2 to 4, wherein the subject has an affected FLC with a serum free light chain (sFLC) level of at least 100 mg / L.

6. The method according to any one of claims 1 to 5, wherein the multiple myeloma is relapsed.

7. The method according to any one of claims 1 to 6, wherein the multiple myeloma is refractory.

8. The method according to any one of claims 1 to 7, wherein the bispecific antibody is administered to the subject intravenously.

9. The method of claim 8, wherein the bispecific antibody is administered to the subject as an infusion.

10. The method of claim 9, wherein the infusion is over a span of 1.5-3 hours.

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