DOSING STRATEGY THAT MITIGATES CYTOKINE RELEASE SYNDROME FOR CD3 / CD20 BIESPECIFIC ANTIBODIES.

Abstract

The present invention relates to administration regimens for therapeutic proteins (e.g., bispecific antibodies that activate T cells) that mitigate cytokine release syndrome and infusion-related reactions. The methods employ an initial fractionated dosing with the optional administration of additional agents such as steroids or cytokine antagonists, which are discontinued at the maximum weekly dosage during the course of the dosing regimen.

Description

DOSING STRATEGY THAT MITIGATES THE RELEASE SYNDROME OF CYTOKINES FOR BISECIFIC ANTIBODIES TO CD3 / CD20 l / U I / DUO REFERENCE TO A LISTING OF SEQUENCES

[0001] This application incorporates by reference the Sequence Listing submitted in computer readable format as file 10496W001-Sequence, created on August 30, 2019 and containing 162,944 bytes. FIELD OF THE INVENTION

[0002] The present invention is in the field of medicine, and relates to dosing strategies and administration regimens of therapeutic antibodies (eg, T cell-directed bispecific antibodies) that mitigate the prevalence and severity of cytokine release syndrome or an infusion-related reaction in patients undergoing immunotherapy. BACKGROUND OF THE INVENTION

[0003] Cytokine release syndrome (CRS) is a systemic inflammatory response that can be triggered by a variety of factors, including certain drugs. Cancer immunotherapies that activate T cells carry a particularly high risk of CRS, which is generally due to target effects induced by binding of a bispecific antibody or T cell chimeric antigen receptor (CAR). to its antigen and subsequent activation of bystander immune cells and non-immune cells, such as endothelial cells. Activation of bystander cells results in the massive release of a series of cytokines. IL-6, IL-10, and interferon (IFN)-γ are some of the major cytokines that are consistently elevated in the serum of CRS patients. With T cell activation therapies directed against tumor cells, CRS is triggered by the massive release of IFN-y by activated T cells or the tumor cells themselves. The secreted IFN-y induces the activation of other immune cells, mainly macrophages, which in turn produce excessive amounts of additional cytokines such as IL-6, TNF-α, and IL-10. IL-6, in particular, contributes to many of the key symptoms of CRS, including vascular leakage, and activation of the complement and coagulation cascade that induces disseminated intravascular coagulation. Furthermore, IL-6 likely contributes to cardiomyopathy by promoting myocardial dysfunction. Shimabukaro-Vornhagen et al., Journal for Immunotherapy of Cancer, 6:56, pp. 1-14, 2018. In some cases, the symptoms associated with CRS are called an infusion-related reaction (IRR) if they occur less than six hours after the start of the infusion, and CRS if they occur more than six hours later. after the start of the infusion. ι / u i rouo

[0004] The management of cancer immunotherapy toxicities is a challenging clinical problem. Mitigation of CRS or IRR is a hallmark of the administration of certain treatment modalities, for example, CAR T cells and T cell-directed bispecific antibodies. Low-grade CRS is usually treated symptomatically with antihistamines, antipyretics and liquids. Severe CRS can represent a life-threatening adverse event that requires prompt and aggressive treatment. The reduction of the tumor load, the limitations in the dose of the administered therapy, and premedication with steroids have reduced the incidence of severe CRS, as has the use of anti-cytokine treatments. Tocilizumab, an anti-IL-6 antibody, has become a standard initial treatment for severe CRS in some circumstances. However, the use of dose and treatment limitations to minimize cytokine activity may have detrimental effects on the efficacy of immunotherapy. Therefore, there remains a need for alternative strategies to mitigate the life-threatening effects of CRS without adversely affecting the therapeutic benefits of immunotherapies. BRIEF DESCRIPTION OF THE INVENTION

[0005] In one aspect, the present invention provides a method of administering a therapeutic protein to a subject in a dosage regimen to mitigate the adverse effects of cytokine release syndrome or infusion-related reaction, comprising: ( i) administering fractions of a primary dose (DI) of the therapeutic protein in week 1 of the dosing regimen, wherein the primary dose comprises no more than 10 mq of the therapeutic protein, a first fraction of the dose (F1D1) comprises 40% to 60% of the total primary dose and is administered to the subject on Day 1 of Week 1, and a second fraction of the dose (F2D1) comprises the remaining 40% to 60% of the total primary dose and is administered to the subject from 12 to 96 hours after the administration of the F1D1; (ii) administering fractions of a secondary dose (D2) of the therapeutic protein in week 2 of the dosing regimen, wherein the secondary dose is not more than one-half of a maximum weekly dose of the therapeutic protein, a first fraction of the dose (F1D2) comprising 40% to 60% of the total secondary dose, a second dose fraction (F2D2) comprising the remaining 40% to 60% of the total secondary dose, and the F2D2 is administered to the subject 12 to 96 hours after F1D2 administration during week 2 of the dosing regimen; and (iii) administering the maximum weekly dose of the therapeutic protein to the subject as a single dose in a subsequent week of the dosing regimen.

[0006] In some cases, the F2D1 is administered to the subject 24 to 96 hours after the F1D1 administration. In some cases, the F2D1 is administered to the subject 18 to 72 hours after the F1D1 administration. In some cases, the F2D2 is administered to the subject 24 to 96 hours after the F1D2 administration. In some cases, the F2D2 is administered to the subject 18 to 72 hours after the F1D2 administration. In some cases, the subsequent week is week 3 of the dosing regimen. In some cases, the subsequent week is week 4 of the dosing regimen. In some cases, the subsequent week is week 14 of the dosing regimen. In some cases, the subsequent week is any of weeks 4 through 36 of the dosing regimen.

[0007] In some embodiments, the method further comprises: (i) administering fractions of a tertiary dose (D3) of the therapeutic protein in week 3 of the dosing regimen, wherein the tertiary dose is not less than half of the maximum weekly dose of the therapeutic protein and not more than the maximum weekly dose of the therapeutic protein, a first dose fraction (F1D3) comprising 40% to 60% of the total tertiary dose, a second dose fraction ( F2D3) comprising the remaining 40% to 60% of the total tertiary dose, and F2D3 is administered to the subject 12 to 96 hours after F1D3 administration during week 3 of the dosing regimen; and (ii) administering the 1 / U I / DUO maximum weekly dose of the therapeutic protein to the subject as a single dose in a subsequent week of the dosing regimen.

[0008] In some cases, the F2D3 is administered to the subject 24 to 96 hours after the F1D3 administration. In some cases, the F2D3 is administered to the subject 18 to 72 hours after the F1D3 administration.

[0009] In some cases, the subsequent week is week 4 of the dosing regimen. In some cases, the subsequent week is week 14 of the dosing regimen. In some cases, the subsequent week is any of weeks 4 through 36 of the dosing regimen. In various modalities, the tertiary dose is administered as a single dose in weeks 4 through 12 of the dosing regimen.

[0010] In one aspect, the present invention provides a method of administering a therapeutic protein to a subject in a dosage regimen to mitigate the adverse effects of cytokine release syndrome or infusion-related reaction, comprising: ( i) administering fractions of a primary dose (DI) of the therapeutic protein in week 1 of the dosing regimen, wherein the primary dose comprises not more than 10 mg of the therapeutic protein, a fraction of the first dose (F1D1) that comprises 40% to 60% of the total primary dose and is administered to the subject on Day 1 of Week 1, and a second dose fraction (F2D1) comprising the remaining 40% to 60% of the total primary dose and it is administered to the subject 12 to 96 hours after the administration of the F1D1; (ii) administering fractions of a secondary dose (D2) of the therapeutic protein in week 2 of the dosing regimen, wherein the secondary dose is equal to a maximum weekly dose of the therapeutic protein, a first fraction of the dose (F1D2 ) comprising 50% of the total secondary dose, a second dose fraction (F2D2) comprising 50% of the total secondary dose, and the F2D2 is administered to the subject 12 to 96 hours after administration of the secondary dose. F1D2 during week 2 of the dosing regimen; and (iii) administering the maximum weekly dose of the therapeutic protein to the subject as a single dose in a subsequent week of the dosing regimen.

[0011] In one aspect, the present invention provides a method of administering a therapeutic protein to a subject in a dosage regimen to mitigate the adverse effects of cytokine release syndrome or infusion-related reaction, comprising: ( i) administering fractions of a primary dose (DI) of the therapeutic protein in week 1 of the dosing regimen, wherein the primary dose comprises no more than 10 mg of the therapeutic protein, wherein DI is administered to the subject in multiple fractions of the dose {for example, F1D1, F2D1, F3D1, F4D1, Λ / C / ZUZ l / U I / DUO F5D1) on subsequent days within week 1; (ii) administering fractions of a secondary dose (D2) of the therapeutic protein in week 2 of the dosing regimen, wherein the secondary dose is equal to or less than the maximum weekly dose of the therapeutic protein, and is administered in multiple fractions (eg, F1D2, F2D2, F3D2, F4D2, F5D2) to the subject on subsequent days within week 2; and (iii) administering the maximum weekly dose (MD) of the therapeutic protein to the subject as multiple fractions of the MD or as a single dose in a subsequent week of the dosing regimen.

[0012] In some embodiments, a second fractional dose (F2) is administered to the subject 12 to 96 (eg, 24 to 72) hours after administration of the first fractional dose (Fl), optionally a third fraction of the dose (F3) is administered to the subject not less than 24 hours after the administration of the second fraction of the dose (F2), optionally a fourth fraction of the dose (F4) is administered to the subject not less than 24 hours after the administration of the third fraction of the dose (F3), and optionally a fifth fraction of the dose (F5) is administered to the subject not less than 24 hours after the administration of the fourth fraction of the dose (F4) during weeks 1, 2, or 3 of the dosing regimen.

[0013] In some embodiments, the methods of the present disclosure further comprise administration of one or more maintenance doses during a maintenance phase of the dosing regimen, which follows completion of a weekly phase of the regimen. In some cases, each maintenance dose is given 2, 3, or 4 weeks after the immediately preceding dose. In one embodiment, the maintenance dose is the maximum weekly dose of the therapeutic protein administered as a single dose.

[0014] In some cases, the maximum weekly dose (MD) of the therapeutic protein is administered to the subject as a single dose for 1-8 weeks, 1-12 weeks, or 1-16 weeks during a weekly phase of the regimen. of dosage. In some instances, the maximum weekly dose of the therapeutic protein is administered to the subject as a single dose once every two weeks during a maintenance phase of the dosing regimen, which follows completion of a weekly phase of the dosing regimen. In some instances, the maximum weekly dose of the therapeutic protein is administered to the subject as a single dose once every three weeks during a maintenance phase of the dosing regimen, which follows completion of a weekly phase of the dosing regimen. In some instances, the maximum weekly dose of the therapeutic protein is administered to the subject as a single dose once every four weeks during a maintenance phase of the dosing regimen, which follows completion of a weekly phase of the dosing regimen. In some modalities, the maintenance phase is a period of up to 86 weeks. In some modalities, the maintenance phase is a period of up to 87 weeks. In some modalities, the maintenance phase is a period of up to 88 weeks. In some modalities, the maintenance phase is greater than 86 weeks, greater than 100 weeks, greater than 150 weeks, greater than 200 weeks, or greater than 250 weeks. In some modalities, the maintenance phase is at least 24 weeks. In some modalities, the maintenance phase is 24 weeks.

[0015] In various embodiments, the primary dose is 1 mg. In various modalities, the secondary dose is 20 mg. In various modalities, the tertiary dose is 40 mg. In various modalities, the tertiary dose is 80 mg. In various modalities, the tertiary dose is 160 mg. In various modalities, the tertiary dose is 320 mg.

[0016] In various embodiments, F1D1 comprises 50% of the total primary dose and F2D1 comprises 50% of the total primary dose. In various embodiments, F1D2 comprises 50% of the total secondary dose and F2D2 comprises 50% of the total secondary dose. In various embodiments, F1D3 comprises 50% of the total tertiary dose and F2D3 comprises 50% of the total tertiary dose.

[0017] In some cases, the maximum weekly dose (MD) A / C / ZUZ l / U I / DUO of therapeutic protein is from 5mg to 320mg. In various modalities, the maximum weekly dose of the therapeutic protein is 6-320 mg, 10-320 mg, 5-40 mg, 5-80 mg, 5-160 mg, 12-40 mg, 18-80 mg, 40- 80mg, 80-160mg, 160-320mg, 5mg, 6mg, 7mg, 8mg, 12mg, 18mg, 27mg, 40mg, 80mg, 160mg, or 320mg. In some modalities, the maximum weekly dose is 80 mg. In some modalities, the maximum weekly dose is 160 mg. In some modalities, the maximum weekly dose is 320 mg. In some cases, the maintenance dose of therapeutic protein is 5 mg to 320 mg. In various modalities, the maintenance dose of therapeutic protein is 6-320mg, 10-320mg, 5-40mg, 5-80mg, 5-160mg, 12-40mg, 18-80mg, 4080mg , 80-160mg, 160-320mg, 5mg, 6mg, 7mg, 8mg, 12mg, 18mg, 27mg, 40mg, 80mg, 160mg, or 320mg. In some modalities, the maintenance dose is 80 mg. In some modalities, the maintenance dose is 160 mg. In some modalities, the maintenance dose is 320 mg.

[0018] In some cases, each dose or fraction of a dose is administered to the subject over a period of 1 to 6 hours.

[0019] In some embodiments, the subject has been diagnosed with a cancer. In some cases, the cancer is a B-cell neoplasm. In some cases, the B-cell neoplasm is a CD20+ B-cell neoplasm. In some cases, the cancer is non-Hodgkin lymphoma, Hodgkin lymphoma, chronic lymphocytic leukemia, acute lymphoblastic leukemia, small lymphocytic lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, marginal zone lymphoma, macroglobulinemia Waldenstrom's lymphoma, primary mediastinal B-cell lymphoma, lymphoblastic lymphoma, or Burkitt's lymphoma. In some cases, the cancer is selected from pancreatic carcinoma, head and neck cancer, prostate cancer, malignant gliomas, osteosarcoma, colorectal cancer, gastric cancer, malignant mesothelioma, multiple myeloma, ovarian cancer, small cell lung cancer, non-small cell lung cancer, synovial sarcoma, thyroid cancer, breast cancer, melanomaglioma, breast cancer, squamous cell carcinoma, esophageal cancer, clear cell renal cell carcinoma, chromophobe cell renal carcinoma, oncocytoma, cell carcinoma transitional tumors, urothelial carcinoma, bladder adenocarcinoma, or small cell bladder carcinoma. In some embodiments, the subject has been diagnosed with follicular lymphoma (LE). In some cases, LE is grade 1-3a. In some embodiments, the subject has been diagnosed with diffuse large B-cell lymphoma (DLBCL). In one embodiment, the subject has been diagnosed with relapsed / refractory GBDL. In some cases, the subject diagnosed with DLBCL has failed prior CAR-T therapy. In some embodiments, the subject has been diagnosed with mantle cell lymphoma (MCL). In some cases, the subject diagnosed with MCL has failed prior Bruton tyrosine kinase (BTK) inhibitor therapy. In some embodiments, the subject has been diagnosed with marginal zone lymphoma (MZL).

[0020] In some cases, the subject is a human, a human adult, or a human child (under the age of eighteen).

[0021] In various embodiments, the therapeutic protein is an antibody or an antigen-binding fragment thereof. In some cases, the antibody is a fully human antibody. In some cases, the antibody is a bispecific antibody or an antigen-binding fragment thereof. In some embodiments, the bispecific antibody or antigen-binding fragment comprises a first antigen-binding arm that binds to a T cell antigen. In some cases, the T cell antigen is CD3. In some cases, the T cell antigen is CD28. In some embodiments, the bispecific antibody or antigen-binding fragment comprises a second antigen-binding arm that binds to an antigen on the tumor cell. In some cases, the tumor cell antigen is selected from the group consisting of AFP, ALK, BAGE, BCMA, BIRC5 (survivin), BIRC7, β-catenin, brc-abl, BRCA1, BORIS, CA9, carbonic anhydrase IX proteins. , caspase-8, CALR, CCR5, CD19, CD20 (MS4A1), CD22, CD40, CD70, CDK4, CEA, cyclin-Bl, CYP1B1, EGFR, Λ / C / ZUZ l / U I / DUO EGFRvIII, ErbB2 / Her2, ErbB3, ErbB4, ETV6-AML, EpCAM, EphA2, Fra-1, FOLR1, GAGE ​​proteins (eg, GAGE-1, -2), GD2, GD3, GloboH, glypican-3, GM3, gplOO, Her2, HLA / B-raf, HLA / kras, HLA / MAGE-A3, hTERT, LMP2, MAGE proteins (eg, MAGE-1, -2, -3, -4, -6, and -12) , MART-1, mesothelin, ML-IAP, Mucl, Muc2, Muc3, Muc4, Muc5, Mucl6 (CA-125), MUM1, NA17, NYBR1, NY-BR62, NY-BR85, NY-ESO1, pl5, p53, PAP, PAX3, PAX5, PCTA-1, PLACI, PRLR, PRAME, PSMA (FOLH1), RAGE proteins, Ras, RGS5, Rho, SART-1, SART-3, STEAP1, STEAP2, TAG-72, TGF-β, TMPRSS2, Thompson-nouvelle (Tn) antigen, TRP-1, TRP-2, tyrosinase, and uroplakin-3.

[0022] In some embodiments, the tumor cell antigen is CD20. In some cases, the bispecific antibody is an anti-CD20 x anti-CD3 antibody. In some cases, the anti-CD20 x anti-CD3 antibody is REGN1979.

[0023] In some embodiments, the tumor cell antigen is BCMA. In some cases, the bispecific antibody is an anti-BCMA x anti-CD3 antibody.

[0024] In some embodiments, the tumor cell antigen is PSMA. In some cases, the bispecific antibody is an anti-PSMA x anti-CD3 antibody.

[0025] In some embodiments, the tumor cell antigen is MUC16. In some cases, the bispecific antibody is an anti-MUC16 x anti-CD3 antibody.

[0026] In some embodiments, the tumor cell antigen is STEAP2. In some cases, the bispecific antibody is an anti-STEAP2 x anti-CD3 antibody.

[0027] In various embodiments, the therapeutic protein is maintained at a serum concentration of approximately 2000 micrograms / liter (mcg / L) or higher after administration of the maximum weekly dose for the duration of the dosing regimen. In some cases, the therapeutic protein is maintained at a serum concentration of approximately 2600 mcg / L or higher after administration of the maximum weekly dose for the duration of the dosing regimen. In some modalities, the therapeutic protein is maintained at a serum concentration of approximately 3700 mcg / L or higher after administration of the maximum weekly dose for the duration of the dosing regimen.

[0028] In some embodiments, the therapeutic protein is administered to the subject in combination with a second agent selected from a steroid, an antihistamine, acetaminophen, a non-steroidal anti-inflammatory drug (ΑΙΝΕ), an IL-6 antagonist, or an antagonist of the IL6R. In some cases, the steroid is dexamethasone. In some cases, the ΑΙΝΕ is indomethacin. In some cases, the IL-6 antagonist is an anti-IL-6 antibody, or the IL-6R antagonist is an anti-IL-6R antibody. In some embodiments, the anti-IL-6R antibody is sarilumab. In A / E / ZUZ I / U I / DUO various modalities, administration of the second agent is eliminated after a first administration of the maximum weekly dose for the duration of the dosing regimen. In other embodiments, the second agent is administered prior to administration of the therapeutic protein (eg, approximately one to three hours prior to F1D1, F2D1, F1D2, F2D2, F1D3, and / or F2D3). In still other embodiments, the therapeutic protein is administered by infusion over a period of time such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more hours.

[0029] In various embodiments, the therapeutic protein is administered to the subject in combination with a second therapeutic agent. In some embodiments, the second therapeutic agent comprises at least one of rituximab, obinutuzumab, cyclophosphamide, doxorubicin, vincristine, prednisone, prednisolone, bendamustine, lenalidomide, chlorambucil, ibritumomab tiuxetan, idelalisib, copanlisib, duvelisib, etoposide, methylprednisolone, cytarabine, cisplatin, mesna, ifosfamide, mitoxantrone, and procarbazine. In some cases, the second therapeutic agent comprises a combination of cyclophosphamide, doxorubicin, vincristine, and prednisone. In some cases, the second therapeutic agent comprises a combination of ifosfamide, cisplatin, and etoposide. In some cases, the second therapeutic agent comprises a combination of gemcitabine and oxaliplatin. In some cases, the second 1 / U I / DUO therapeutic agent comprises a combination of lenalidomide and rituximab. In some cases, the second therapeutic agent is lenalidomide.

[0030] In one aspect, the present invention includes a method of treating a B-cell cancer in a subject, comprising: (a) selecting a subject diagnosed with a B-cell cancer; and (b) administering a therapeutic protein to the subject according to any of the methods discussed above or herein using a dosage regimen to mitigate the adverse effects of cytokine release syndrome or infusion related reaction. In some embodiments, the subject has previously been treated with anti-CD20 antibody therapy. In some embodiments, the subject has previously been treated with a CAR-T therapy. In some cases, the B-cell cancer is selected from the group consisting of follicular lymphoma, diffuse large B-cell lymphoma, mantle cell lymphoma, and marginal zone lymphoma.

[0031] In any of the modalities discussed above or herein, the incidence of CRS and grade 3 IRR is less than 10%. In some cases, the incidence of CRS and grade 3 IRR is less than 7.5% or less than 7%. In some modalities, the maximum weekly dose is 80 mg or higher when the incidence of CRS and the IRR is less than 10%, less than 9%, less than 8%, less than 7.5% or less than 7%. In either modality, any dose administered as a single dose can be administered in no more than 1 hour.

[0032] In various embodiments, any of the features or components of any of the embodiments discussed above or herein may be combined, and such combinations are included within the scope of the present disclosure. Any specific value discussed above or herein may be combined with another related value discussed above or herein to cite a range with values ​​representing the upper and lower extremes of the range, and such ranges are included within the scope of the present description. A therapeutic protein for use in any of the methods discussed in the present description, or the use of a therapeutic protein in the manufacture of a medicament for use in any of the methods discussed in the present description, are also included within the scope. of this description.

[0033] Other embodiments will be apparent from a review of the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 illustrates the incidence of CRS / IRR during the first five weeks of REGN1979 therapy. A / E / ZUZ l / U I / DUO at various dose levels. DETAILED DESCRIPTION

[0035] Before describing the present invention, it should be understood that this invention is not limited to the particular methods and experimental conditions described, as these methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, as the scope of the present invention shall be limited only by the appended claims.

[0036] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as is commonly understood by a person skilled in the art to which this invention pertains. The term approximately, when used in reference to a particular stated numerical value, means that the value may differ from the stated value by no more than 1%. For example, the expression about 100 includes 99 and 101 and all values ​​in between (for example, 99.1, 99.2, 99.3, 99.4, etc.) .

[0037] Although any methods and materials similar or equivalent to those described herein may be used in the practice or testing of the present invention, preferred methods and materials are now described. All patents, applications and non-patent publications mentioned in this specification are incorporated herein by reference in their entirety.

[0038] The expression CD3 refers to an antigen that is expressed on T cells as part of the T cell multimolecular receptor (TCR) and that consists of a homodimer or heterodimer formed from the association of two of four chains. receptors: CD3-epsilon, CD3-delta, CD3zeta, and CD3-gamma. Human CD3-epsilon comprises the amino acid sequence as set forth in SEQ ID NO: 1; human CD3-delta comprises the amino acid sequence as set forth in SEQ ID NO: 2; human CD3-zeta comprises the amino acid sequence as set forth in SEQ ID NO: 3; and CD3-gamma comprises the amino acid sequence as set forth in SEQ ID NO 4.

[0039] All references to proteins, polypeptides, and protein fragments herein are intended to refer to the human version of the respective protein, polypeptide, or protein fragment, unless explicitly specified as being from a non-human species. Thus, the term "CD3" means human CD3 unless specified as coming from a non-human species, eg, mouse CD3, monkey CD3, etc.

[0040] An antigen-binding domain that binds to CD3, - an antibody that binds to CD3 or an anti-CD3 antibody Λ / C / ZUZ I / U I / DUO includes antibodies and antigen-binding fragments thereof that specifically recognize a single CD3 subunit (eg, epsilon, delta, gamma, or zeta), as well as antibodies and fragments antigen-binding proteins thereof that specifically recognize a dimeric complex of two CD3 subunits (eg, CD3 gamma / epsilon, delta / epsilon, and zeta / zeta dimers). Antibodies and antigen-binding fragments of the present invention can bind to soluble CD3 and / or CD3 expressed on the cell surface. Soluble CD3 includes the natural CD3 proteins, as well as recombinant variants of the CD3 protein, such as, for example, monomeric and dimeric CD3 constructs that lack a transmembrane domain or are not otherwise associated with a cell membrane.

[0041] The expression CD20, refers to a non-glycosylated phosphoprotein expressed on the cell membranes of mature B cells. CD20 is considered a B-cell tumor-associated antigen because it is expressed on more than 95% of B-cell non-Hodgkin lymphomas (NHL) and other B-cell neoplasms, but it is absent on B-cell precursors, B cells. dendritic and plasma cells. The human CD20 protein has the amino acid sequence shown in SEQ ID NO:5.

[0042] An antigen-binding domain that binds CD20, an antibody that binds CD20, or an anti-CD20 antibody. A / E / ZUZ l / U I / DUO CD20 includes antibodies and antigen-binding fragments thereof that specifically recognize CD20.

[0043] The expression BCMA, refers to the B cell maturation antigen. BCMA (also known as TNFRSF17 and CD269) is a cell surface protein expressed on malignant plasma cells, and plays a central role in the regulation of the maturation and differentiation of B cells into immunoglobulin-producing plasma cells. The amino acid sequence of human BCMA is shown in SEQ ID NO: 6.

[0044] A BCMA-binding antigen-binding domain, a BCMA-binding antibody or an anti-BCMA antibody includes those antibodies and antigen-binding fragments thereof that specifically recognize BCMA.

[0045] The term PSMA refers to the prostate-specific membrane antigen, also known as folate hydrolase 1 (FOLH1). PSMA is a non-separable integral membrane glycoprotein that is highly expressed in prostate epithelial cells and is a cell surface marker of prostate cancer. The amino acid sequence of human PSMA is as set forth in SEQ ID NO: 7.

[0046] An antigen-binding domain that binds PSMA, an antibody that binds PSMA, or an anti- A / E / ZUZ l / U I / DUO PSMA includes those antibodies and antigen-binding fragments thereof that specifically recognize PSMA.

[0047] The expression MUC16, refers to mucin 16. MUC16 is a single transmembrane domain of highly glycosylated integral membrane glycoprotein that is highly expressed in ovarian cancer. The amino acid sequence of human MUC16 is as set forth in SEQ ID NO: 8.

[0048] A MUC16-binding antigen-binding domain, an antibody that binds MUC16 or an anti-MUC16 antibody includes those antibodies and antigen-binding fragments thereof that specifically recognize MUC16.

[0049] The expression STEAP2 refers to an epithelial antigen of six transmembrane segments of the prostate 2. STEAP2 is an integral protein that spans six transmembrane segments, which is highly expressed in the epithelial cells of the prostate and is a cell surface marker for prostate cancer. STEAP2 is a 490 amino acid protein encoded by the STEAP2 gene located on chromosome region 7q21 in humans. The amino acid sequence of human STEAP2 is set forth in SEQ ID NO: 9.

[0050] An antigen-binding domain that binds STEAP2, an antibody that binds STEAP2, or an anti-STEAP2 antibody includes those antibodies and antigen-binding fragments thereof that specifically recognize STEAP2. A / E / ZUZ l / U I / DUO

[0051] The term "therapeutic protein" includes any polypeptide, including antibodies and antigen-binding fragments thereof, and bispecific antibodies and antigen-binding fragments thereof, that are used to prevent, treat, or ameliorate any condition, disease or disorder in a subject.

[0052] The term "antigen-binding molecule" includes antibodies and antigen-binding fragments of antibodies, including, for example, bispecific antibodies.

[0053] The term "antibody" means any antigen-binding molecule or molecular complex comprising at least one complementarity-determining region (CDR) that specifically binds to or interacts with a particular antigen (for example, CD20, BCMA , PSMA, MUC16, STEAP2 or CD3). The term antibody includes immunoglobulin molecules that comprise four polypeptide chains, two heavy (H) and two light (L) chains interconnected by disulfide bonds, as well as their multimers (eg, IgM). The term antibody also includes immunoglobulin molecules that consist of four polypeptide chains, two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or Vh) and A / E / ZUZ l / U I / DUO a constant region of the heavy chain. The heavy chain constant region comprises three domains, ChI, Ch2 and Ch3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or Vl) and a light chain constant region. The constant region of the light chain comprises a domain (ClI). The Vh and Vl regions can also be subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with more conserved regions, termed framework regions (FR). Each Vh and Vl is composed of three CDRs and four FRs, arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In different embodiments of the invention, the FRs of the antibody (or the antigen-binding portion thereof) may be identical to human germline sequences, or may be naturally or artificially modified. A consensus amino acid sequence can be defined based on a parallel analysis of two or more CDRs. The term antibody includes a bispecific antibody unless otherwise indicated.

[0054] The term "antibody" also includes antigen-binding fragments of full-length antibody molecules. The terms antigen-binding portion of an antibody, antigen-binding fragment of an antibody, and the like, include any naturally occurring, enzymatically derived, synthetic, or genetically engineered I / U I / DUO polypeptide or glycoprotein that specifically binds to an antigen to form a complex. Antigen-binding fragments of an antibody can be derived, for example, from complete antibody molecules using any suitable standard technique, such as proteolytic digestion, or recombinant genetic engineering techniques involving DNA manipulation and expression. encoding variable and, optionally, constant antibody domains. Such DNA is known and / or readily available, for example, from commercial sources, DNA libraries (including, for example, phage-antibody libraries), or can be synthesized. DNA can be sequenced and manipulated chemically or through the use of molecular biology techniques, for example, to arrange one or more variable and / or constant domains in a suitable configuration, or to introduce codons, create tanker residues, modify, add or remove amino acids, etc.

[0055] The term "bispecific antigen-binding molecule" refers to a protein, polypeptide, or molecular complex comprising at least a first antigen-binding domain and a second antigen-binding domain. Each antigen-binding domain within the bispecific antigen-binding molecule comprises at least one CDR that, alone or in combination with one or more additional CDRs and / or FR ι / u i rouo, specifically binds to a particular antigen. . Bispecific antigen binding molecules include bispecific antibodies.

[0056] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units consisting of amino acid residues that mimic the hypervariable region of an antibody (for example, an isolated complementarity determining region (CDR), such as a CDR3 peptide), or a FR3CDR3- peptide. FR4 restricted. Other engineered molecules, such as domain-specific antibodies, single domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies , etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains, are also included within the term antigen-binding fragment.

[0057] An antigen-binding fragment of an antibody will typically comprise at least one variable domain. The variable domain can be of any size or amino acid composition, and generally comprises at least A / E / ZUZ l / U I / DUO a CDR that is adjacent to or in frame with one or more frame sequences. In antigen-binding fragments having a Vh domain associated with a Vl domain, the VH and VL domains may be positioned relative to each other in any suitable arrangement. For example, the variable region can be dimeric and contain Vh-Vh, Vh-Vl or Vl-Vl dimers. Alternatively, the antigen-binding fragment of an antibody may contain a monomeric Vh or Vl domain.

[0058] In certain embodiments, an antigen-binding fragment of an antibody may contain at least one variable domain covalently linked to at least one constant domain. Non-limiting examples of constant and variable domain configurations that can be found within an antigen-binding moiety of an antibody of the present invention include: (i) Vh-Ch1; (ü) Vh-Ch2; (iii) hCh3; (iv) Vh-Ch1-Ch2; (v) Vh-Ch1-Ch2-Ch3 ; (vi) Vh-Ch2-Ch3; (vii) Vh-Cl; (viii) Vl-Ch1; (ix) Vl-Ch2; (x) Vl-Ch3; (xi) Vl-Ch1-Ch2; (xii) Vl-Ch1-Ch2-Ch3 ; (xiii) Vl-Ch2-Ch3; and (xiv) VL-CL. In any configuration of the variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains may be directly linked to each other or may be linked by a full or partial hinge region or linker region. A hinge region may consist of at least 2 {for example, 5, 10, 15, 20, 40, 60 or more) amino acids that result in flexible or semi-flexible linkage between adjacent variable and / or constant domains in a single molecule. of polypeptides. In addition, an antigen-binding fragment of an antibody of the present invention may comprise a homodimer or heterodimer (or other multimer) of any of the above-listed constant and variable domain configurations in non-covalent association with each other and / or with one or more monomeric Vh or Vl domains (eg, via disulfide bonds).

[0059] Like full-length antibody molecules, antigen-binding fragments can be monospecific or multispecific (eg, bispecific). A multispecific antigen-binding fragment of an antibody will typically comprise at least two distinct variable domains, each variable domain being capable of specifically binding to a separate antigen or to a distinct epitope on the same antigen. Any multispecific antibody format, including the exemplary bispecific antibody formats described herein, can be adapted for use in the context of an antigen-binding fragment of an antibody of the present invention using available routine techniques. in technique.

[0060] The antibodies of the present invention may function through the cytotoxicity dependent on the A / E / ZUZ l / U I / DUO complement (CDC) or antibody-dependent cell-mediated cytotoxicity (ADCC). Complement-dependent cytotoxicity (CDC) refers to the lysis of cells expressing antigen by an antibody of the invention in the presence of complement. Antibody-dependent cell-mediated cytotoxicity (ADCC) refers to a cell-mediated reaction in which non-specific cytotoxic cells expressing Fe receptors (FcR) (for example, Natural Killer (NK) cells, neutrophils, and macrophages) recognize the antibody. bound antibody on a target cell and thus lead to lysis of the target cell. CDC and ADCC can be measured using assays that are well known and available in the art. (See, eg, US Patent Nos. 5,500,362 and 5,821,337, and Clynes et al. (1998) Proc. Nati. Acad. Sci. (USA) 95:652656). The constant region of an antibody is important in the ability of an antibody to fix complement and mediate cell-dependent cytotoxicity. Therefore, the isotype of an antibody can be selected on the basis of whether it is desirable for the antibody to mediate cytotoxicity. The antibodies of the present disclosure may include a human IgG heavy chain. In various embodiments, the weight may be of the IgG1, IgG2, IgG3, or IgG4 isotype.

[0061] In certain embodiments of the invention, the antibodies or bispecific antibodies are human antibodies. The term "human antibody" is intended to include antibodies having variable and constant regions derived from human germ line immunoglobulin sequences. Human antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (for example, mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo), for example, in the CDRs and in particular the CDR3. The term "human antibody," however, is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto the human framework sequences.

[0062] In some embodiments, the antibodies of the invention may be recombinant human antibodies. The term "recombinant human antibody" is intended to include all human antibodies that are prepared, expressed, created, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell (described below), antibodies isolated from a recombinant combinatorial human antibody library (described below), antibodies isolated from an animal (eg, mouse) that is transgenic for human immunoglobulin genes (see, eg, Taylor and others (1992) Nuci.Acids A / E / ZUZ l / U I / DUO Res. 20:6287-6295) or antibodies prepared, expressed, created, or isolated by any other means that involves splicing human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germinal immunoglobulin sequences. In certain embodiments, however, such recombinant human antibodies are subjected to in vitro mutagenesis (or, when a transgenic animal is used for human Ig sequences, in vivo somatic mutagenesis), and thus the amino acid sequences of the regions Vh and Vl of the recombinant antibodies are sequences which, although derived from and related to human germline Vh and Vl sequences, may not naturally exist in the germline repertoire of human antibodies in vivo.

[0063] Human antibodies can exist in two forms that are associated with hinge heterogeneity. In one form, an immunoglobulin molecule comprises a stable four-chain construct of approximately 150-160 kDa in which the dimers are held together by an interchain heavy chain disulfide bond. In a second form, the dimers are not linked via interchain disulfide bonds and a molecule of approximately 75-80 kDa is formed composed of a covalently coupled heavy and light chain (half antibody). These forms have been extremely difficult to separate, even after affinity purification.

[0064] The frequency of occurrence of the second form in various intact IgG isotypes is due to, but not limited to, structural differences associated with the isotype of the antibody hinge region. A single amino acid substitution in the hinge region of the human IgG4 hinge can significantly reduce the occurrence of the second form (Angal et al. (1993) Molecular Immunology 30:105) to levels typically observed using a hinge hinge. human IgGl. The present invention encompasses antibodies having one or more mutations in the hinge, Ch2, or Ch3 region that may be desirable, eg, in production, to improve the yield of the desired antibody form.

[0065] The antibodies of the invention may be isolated antibodies. An isolated antibody means an antibody that has been identified and separated, and / or recovered from at least one component of its natural environment. For example, an antibody that has been separated or eliminated from at least one component of an organism, or from a tissue or cell in which the antibody naturally occurs or occurs naturally, is an isolated antibody for purposes of the present invention. An isolated antibody also includes an antibody in situ within a recombinant cell. Isolated antibodies are antibodies that have been subjected to at least one purification or isolation step. According to certain embodiments, an isolated antibody can be substantially free of other cellular materials and / or chemicals.

[0066] The antibodies discussed herein may comprise one or more amino acid substitutions, insertions, and / or deletions in the framework and / or CDR regions of the heavy and light chain variable domains compared to germline sequences. from which the antibodies were derived. Such mutations can be readily determined by comparing the amino acid sequences described herein with germline sequences available in, for example, public antibody sequence databases. The present invention also includes antibodies and antigen-binding fragments thereof, which are derived from any of the amino acid sequences described herein, wherein one or more amino acids within one or more framework regions and / or CDR regions are mutated to the corresponding residues of the germline sequence from which the antibody is derived, or to corresponding residues of another human germline sequence, or to a conservative amino acid substitution of the corresponding germline residues (such germline changes sequence are referred to herein collectively as line mutations A / E / ZUZ l / U I / DUO germinal). A person skilled in the art can readily produce, from the heavy and light chain variable region sequences described herein, numerous antibodies and antigen-binding fragments comprising one or more individual germline or combinations thereof. In certain embodiments, all framework and / or CDR residues within the Vh and / or Vl domains are mutated back to residues found in the original germline sequence from which the antibody was derived. In other embodiments, only certain residues are mutated back to the original germline sequence, for example, only mutated residues found within the first 8 amino acids of FR1 or within the last 8 amino acids of FR4, or only mutated residues found within the first 8 amino acids of FR4. within CDR1, CDR2 or CDR3. In other embodiments, one or more of the framework and / or CDR residues are mutated to corresponding residues of a different germline sequence (i.e., a germline sequence that is different from the germline sequence of the which the antibody was originally derived). In addition, the antibodies of the present invention may contain any combination of two or more germline mutations in the framework and / or CDR regions, for example, where certain individual residues are mutated to the corresponding residue of a particular germline sequence. while other 1 / U I / DUO residues that differ from the original germline sequence are maintained or mutated to the corresponding residue of a different germline sequence. Once obtained, antibodies and antigen-binding fragments containing one or more germline mutations can be readily assayed for one or more desired properties, such as improved binding specificity, increased binding affinity, biological properties enhanced agonists or antagonists (as the case may be), reduced immunogenicity, etc. Antibodies and antigen-binding fragments obtained in this general manner are included within the present invention.

[0067] The present invention also includes antibodies comprising variants of any of the HCVR, LCVR and / or CDR amino acid sequences described herein having one or more conservative substitutions. For example, the present invention includes antibodies having HCVR, LCVR, and / or CDR amino acid sequences with, for example, 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, etc., conservative amino acid substitutions. with respect to any of the HCVR, LCVR and / or CDR amino acid sequences set forth herein, the anti-CD3 antibodies described in WO 2014 / 047231 or WO 2017 / 053856, the anti-CD20 x anti- CD3 described in the document WO 2014 / 047231, the anti-PSMA antibodies or A / C / ZUZ l / U I / DUO anti-PSMA x anti-CD3 described in document WO 2017 / 023761, the anti-MUC16 or anti-MUC16 x anti-CD3 antibodies described in document WO 2018 / 067331, the antibodies anti-STEAP2 or anti-STEAP2 x anti-CD3 described in WO 2018 / 058001, or the anti-BCMA or anti-BCMA x antiCD3 antibodies described in US 62 / 700,596 (filed July 19, 2018), each of which is incorporated herein by reference.

[0068] The term "epitope" refers to an antigenic determinant that interacts with a specific antigen-binding site in the variable region of an antibody molecule known as a paratope. A single antigen may have more than one epitope. Therefore, different antibodies can bind to different areas on an antigen and can have different biological effects. Epitopes can be conformational or linear. A conformational epitope is produced by the spatially juxtaposed amino acids of different segments of the linear polypeptide chain. A linear epitope is one produced by adjacent amino acid residues in a polypeptide chain. Under certain circumstances, an epitope may include saccharide residues, phosphoryl groups, or sulfonyl groups on the antigen.

[0069] The term substantially identical or substantially identical, when referring to a nucleic acid or fragment thereof, indicates that, when l / U I / DUO are optimally aligned with appropriate nucleotide insertions or deletions or with another nucleic acid (or its complementary strand), there is at least about 95%, and more preferably, at least about 96, 97, 98, or 99% nucleotide sequence identity, as measured by any well-known algorithm sequence identity, such as FASTA, BLAST or Gap, as explained below. A nucleic acid molecule having substantial identity with a reference nucleic acid molecule can, in some cases, encode a polypeptide that has an identical or substantially similar amino acid sequence to the polypeptide encoded by the reference nucleic acid molecule.

[0070] As applied to polypeptides, the term "substantially similar" or "substantially similar" means that two peptide sequences, when optimally aligned, such as by the GAP or BESTFIT programs using predetermined gap weights, share at least 95% share. % sequence identity, even more preferably, at least 98% or 99% sequence identity. Preferably, non-identical residue positions differ by conservative amino acid substitutions. A conservative amino acid substitution is one in which one amino acid residue is replaced by another amino acid residue that has a side chain (group l / U I / DUO R) with similar chemical properties (for example, charge or hydrophobicity). In general, a conservative amino acid substitution will not substantially modify the functional properties of a protein. In cases where two or more amino acid sequences differ from one another by conservative substitutions, the percentage sequence identity or degree of similarity can be adjusted upward to correct for the conservative nature of the substitution. Means for making this adjustment are well known to those skilled in the art. See, for example, Pearson (1994) Methods Mol. Biol. 24: 307-331, incorporated herein by reference. Examples of groups of amino acids that have side chains with similar chemical properties include (1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; (2) aliphatic hydroxyl side chains: serine and threonine; (3) amide-containing side chains: asparagine and glutamine; (4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; (5) basic side chains: lysine, arginine, and histidine; (6) acidic side chains: aspartate and glutamate, and (7) sulfur-containing side chains are cysteine ​​and methionine. Preferred conservative amino acid substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysinearginine, alanine-valine, glutamate-aspartate, and asparaginal / U I / DUO glutamine. Alternatively, a conservative replacement is any change that has a positive value in the PAM250 log-likelihood matrix described in Gonnet et al. (1992) Science 256: 1443-1445, incorporated herein by reference. A moderately conservative replacement is any change that has a non-negative value in the PAM250 log-likelihood matrix.

[0071] Sequence similarity for polypeptides, which is also known as sequence identity, is typically measured using sequence analysis software. Protein analysis software matches similar sequences using measures of similarity assigned to various substitutions, deletions, and other modifications, including conservative amino acid substitutions. For example, the GCG software contains programs, such as Gap and Bestfit, which can be used with predetermined parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species of organisms or between a wild-type protein and a mutein thereof. See, for example, GCG Version 6.1. Polypeptide sequences can also be compared using FASTA using default or recommended parameters, a program in GCG Version 6.1. FASTA (for example, FASTA2 and FASTA3) provides alignments and percentages of A / E / ZUZ l / U I / DUO sequence identity of the regions of best overlap between the unknown sequence and the search sequence (Pearson (2000) supra). Another preferred algorithm when comparing a sequence of the invention to a database containing a large number of sequences from different organisms is the BLAST computer program, especially BLASTP or TBLASTN, using predetermined parameters. See, for example, Altschul et al. (1990) J. Mol. Biol. 215:403-410 and Altschul et al. (1997) Nucleic Acids Res. 25:3389-402, each incorporated herein by reference. Dosing Strategies and Administration Regimens

[0072] Dosing strategies have been developed that provide administration regimens to mitigate the prevalence or severity, or both, of cytokine release syndrome (CRS) or infusion related reaction (IRR) by administration of a protein treatment to a patient for various therapies. In accordance with certain embodiments of the present invention, these strategies include multiple doses of a therapeutic protein or antigen-binding molecule (eg, an antibody or a bispecific antibody) that may be administered to a subject over a defined period of time to create a regime. Methods according to this aspect of the invention comprise sequential administration to a subject A / E / ZUZ l / U I / DUO of multiple doses of a therapeutic protein of the invention. Sequential administration means that each dose of a therapeutic protein is administered to the subject at a different time point, eg, on different days separated by a predetermined interval (eg, hours, days, weeks, or months). The present invention includes methods comprising sequentially administering to the patient a divided primary dose of a therapeutic protein, followed by a divided secondary dose of the therapeutic protein, optionally followed by a divided tertiary dose of the therapeutic protein, followed by single doses of the maximum weekly dose of the therapeutic protein. The present administration regimens allow for higher doses of the therapeutic protein that are desirable to enhance therapeutic efficacy, but without the deleterious effects associated with CRS or IRR. Without wishing to be bound by any particular theory, the present administration regimens provide priming of the immune response to therapeutic protein administration to minimize the incidence and severity of CRS and IRR during the initial phases of the treatment regimen, thus it then allows administration of higher doses of the therapeutic proteins during later phases of the treatment regimen without significant adverse events associated with CRS or IRR.

[0073] An exemplary administration regimen includes: (i) administering fractions of a primary dose of the therapeutic protein in week 1 (SI) of the dosing regimen, wherein the primary dose comprises not more than 1 mg of the therapeutic protein, a first fraction of the dose (F1D1) comprises 50% of the primary dose and is administered to the subject on Day 1 of Week 1, and a second dose fraction (F2D1) comprises 50% of the total primary dose and is administered to the subject within 96 hours following the administration of F1D1; (ii) administering fractions of a secondary dose of the therapeutic protein in week 2 (W2) of the dosing regimen, wherein the secondary dose is not more than one-half of a maximum weekly dose of the therapeutic protein, a fraction of the The first dose (F1D2) comprises 50% of the secondary dose, a second dose fraction (F2D2) comprises 50% of the secondary dose, and F1D2 and F2D2 are administered to the subject 96 hours apart for week 2 of the dosing regimen; and (iii) administering the maximum weekly dose of the therapeutic protein to the subject as a single dose in a subsequent week (Ss) of the dosing regimen.

[0074] Another exemplary administration regimen includes: (i) administering fractions of a primary dose of the therapeutic protein in week 1 (SI) of the dosing regimen, wherein the primary dose comprises no more than 1 mg of the therapeutic protein , a first fraction of the dose A / E / ZUZ 1 / U I / DUO (F1D1) comprises 50% of the primary dose and is administered to the subject on Day 1 of Week 1, and a second fraction of the dose (F2D1) comprises 50% of the total primary dose and is administered to the subject within 96 hours of F1D1 administration; (ii) administering fractions of a secondary dose of the therapeutic protein in week 2 (W2) of the dosing regimen, wherein the secondary dose is not more than one-half of a maximum weekly dose of the therapeutic protein, a fraction of the The first dose (F1D2) comprises 50% of the secondary dose, a second dose fraction (F2D2) comprises 50% of the secondary dose, and F1D2 and F2D2 are administered to the subject 96 hours apart for week 2 of the dosing regimen; (iii) administering fractions of a tertiary dose of the therapeutic protein in week 3 (W3) of the dosing regimen, wherein the tertiary dose is not less than half the maximum weekly dose of the therapeutic protein and not more than the maximum weekly dose of therapeutic protein, a first dose fraction (F1D3) comprises 50% of the tertiary dose, a second dose fraction (F2D3) comprises 50% of the tertiary dose, and F1D3 and F2D3 are administered to the subject within 96 hours of each other during week 3 of the dosing regimen; and (iv) administering the maximum weekly dose of the therapeutic protein to the subject as a single dose in a subsequent week (Ss). A / E / ZUZ l / U I / DUO of the dosage regimen.

[0075] In various embodiments, the primary dose of the therapeutic protein can range from 0.1 mg to 10 mg or more. In some cases, the primary dose of the therapeutic protein is 0.5 mg to 10 mg, 1-10 mg, 2-5 mg, or 5-10 mg. In some cases, the primary dose of the therapeutic protein is 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10mg. In various embodiments, the primary dose of the therapeutic protein comprises a range that includes any of the values ​​noted above as the upper or lower end of the range (eg, 1-5 mg).

[0076] In various modalities, the maximum weekly dose of the therapeutic protein is 5 mg to 320 mg. In some modalities, the maximum weekly dose of the therapeutic protein is from more than 5 mg up to 320 mg. In some cases, the maximum weekly dose of the therapeutic protein is 6-320 mg, 10-320 mg, 5-40 mg, 5-80 mg, 5-160 mg, 12-40 mg, 18-80 mg, 40-80 mg, 80-160 mg, 160-320 mg, 5 mg, 6 mg, 7 mg, 8 mg, 12 mg, 18 mg, 27 mg, 40 mg, 80 mg, 160 mg, or 320 mg. In some cases, the maximum weekly dose of therapeutic protein is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 , 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 45, 50, 55, 60, 65, 70, 75, 80 , 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, A / E / ZUZ l / U I / DUO 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, or 320 mg. In various embodiments, the maximum weekly dose of the therapeutic protein comprises a range that includes any of the values ​​indicated above as the upper or lower end of the range {eg, 200-300 mq).

[0077] In various modalities, the secondary dose comprises 50% of the maximum weekly dose of the therapeutic protein. In some cases, the secondary dose comprises 49%, 48%, 47%, 46%, 45%, 44%, 43%, 42%, 41%, 40%, 39%, 38%, 37%, 36% , 35%, 34%, 33%, 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19 %, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, or 10% of the maximum weekly dose of the therapeutic protein. In some cases, the secondary dose of the therapeutic protein comprises a percentage range of the maximum weekly dose that includes any of the values ​​indicated above as the upper or lower end of the range {eg, 35-50

[0078] In various modalities, the tertiary dose comprises 50% of the maximum weekly dose of the therapeutic protein. In some cases, the tertiary dose comprises 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64% , 65%, 66%, 67%, 68%, 69%, 70%, 71%, t l / U I / DUO%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% of the maximum weekly dose of therapeutic protein. In some cases, the tertiary dose of the therapeutic protein comprises a percentage range of the maximum weekly dose that includes any of the values ​​indicated above as the upper or lower end of the range {eg, 50-75

[0079] In various embodiments, the first dose fraction and the second dose fraction of the primary, secondary and / or tertiary doses each comprise 50% of the dose. In some cases, the first dose fraction and the second dose fraction of the primary, secondary and / or tertiary doses comprise different percentages (totaling to 100%) of the total dose. For example, the first dose fraction may comprise 45% of the dose, and the second dose fraction may comprise 55% of the dose. Alternatively, the first dose fraction may comprise 55% of the dose, and the second dose fraction may comprise 45% of the dose. In various embodiments, the first and second fractions of the dose may include 10 / 90%, 15 / 85%, 20 / 80%, 25 / 75%, 30 / 70%, 35 / 65%. , 40% / 60%, 46% / 54%, 47% / 53%, 48% / 52%, 49% / 51%, or vice versa, of the total primary, secondary, or tertiary dose, respectively.

[0080] In various embodiments, the primary, secondary, and / or tertiary doses (DI, D2, and / or D3) may be divided into two or more fractions. Several options for dividing doses into two fractions are described above. In some cases, however, the doses are divided into 3, 4, or 5 fractions. For example, the primary dose could be divided into 5 fractions, each comprising 20% ​​of the total primary dose, and each dose fraction (F1D1, F2D1, F3D1, F4D1, and F5D1) may be administered to the subject in on five consecutive days during the first week of the dosing regimen. In other cases, the percentage of the total dose (eg, the primary dose) may vary between each fraction of the dose. For example, if the primary dose is divided into 3 fractions, the first dose fraction (F1D1) may include 30% of the total primary dose, the second dose fraction (F2D1) may include 30% of the dose total primary dose, and the third fraction of the dose (F3D1) may include the remaining 40% of the total primary dose. Other combinations of percentages and number of fractional doses that equal 100% of the total dose are expressly contemplated herein.

[0081] In an exemplary embodiment of the dosing regimen, the primary dose comprises 1 mg, the first dose fraction (F1D1) and the second dose fraction (F2D1) each comprise 500 mcg, the secondary dose comprises 6 mg, each of the first dose fraction (F1D2) and the second dose fraction (F2D2) comprises 3 mg, the tertiary dose comprises 12 mg, each of the first dose fraction (F1D3) and the second fraction of the dose (F2D3) comprises 6 mg, and the maximum weekly dose comprises 12 mg of the therapeutic protein.

[0082] In an exemplary embodiment of the dosing regimen, the primary dose comprises 1 mg, the first dose fraction (F1D1) and the second dose fraction (F2D1) each comprise 500 mcg, the secondary dose comprises 9 mg, each of the first dose fraction (F1D2) and the second dose fraction (F2D2) comprises 4.5 mg, the tertiary dose comprises 18 mg, each of the first dose fraction (F1D3) and the second fraction of the dose F2D3) comprises 9 mg, and the maximum weekly dose comprises 18 mg of the therapeutic protein.

[0083] In an exemplary embodiment of the dosing regimen, the primary dose comprises 1 mg, the first dose fraction (F1D1) and the second dose fraction (F2D1) each comprise 500 mcg, the secondary dose comprises 13.5 mg, each of the first dose fraction (F1D2) and the second dose fraction (F2D2) comprises 6.75 mg, the tertiary dose comprises 27 mg, each of the first dose fraction (F1D3) and the second fraction of the F2D3 dose) comprises 13.5 mg, and the maximum weekly dose comprises mg of the therapeutic protein.

[0084] In an exemplary embodiment of the dosing regimen, the primary dose comprises 1 mg, the first dose fraction (F1D1) and the second dose fraction (F2D1) each comprise 500 mcg, the secondary dose comprises 20 mg, each of the first dose fraction (F1D2) and the second dose fraction (F2D2) comprises 10 mg, the tertiary dose comprises 40 mg, each of the first dose fraction (F1D3) and the second fraction of the dose F2D3) comprises 20 mg, and the maximum weekly dose comprises 40 mg of the therapeutic protein.

[0085] In an exemplary embodiment of the dosing regimen, the primary dose comprises 1 mg, the first dose fraction (F1D1) and the second dose fraction (F2D1) each comprise 500 mcg, the secondary dose comprises 20 mg, each of the first dose fraction (F1D2) and the second dose fraction (F2D2) comprises 10 mg, the tertiary dose comprises 60 mg, each of the first dose fraction (F1D3) and the second fraction of the dose F2D3) comprises 30 mg, and the maximum weekly dose comprises 80 mg of the therapeutic protein.

[0086] In an exemplary embodiment of the dosing regimen, the primary dose comprises 1 mg, the first dose fraction (F1D1) and the second dose fraction (F2D1) each comprise 500 mcg, the secondary dose l / U I / DUO comprises 20 mg, each of the first fractional dose (F1D2) and the second fractional dose (F2D2) comprises 10 mg, the tertiary dose comprises 80 mg, each of the first fractional dose ( F1D3) and the second fraction of the dose F2D3) comprises 40 mg, and the maximum weekly dose comprises 160 mg of therapeutic protein.

[0087] In an exemplary embodiment of the dosing regimen, the primary dose comprises 1 mg, the first dose fraction (F1D1) and the second dose fraction (F2D1) each comprise 500 mcg, the secondary dose comprises 20 mg, each of the first dose fraction (F1D2) and the second dose fraction (F2D2) comprises 10 mg, the tertiary dose comprises 120 mg, each of the first dose fraction (F1D3) and the second fraction of the F2D3 dose) comprises 60 mg, and the maximum weekly dose comprises 240 mg of therapeutic protein.

[0088] In an exemplary embodiment of the dosing regimen, the primary dose comprises 1 mg, the first dose fraction (F1D1) and the second dose fraction (F2D1) each comprise 500 mcg, the secondary dose comprises 20 mg, each of the first dose fraction (F1D2) and the second dose fraction (F2D2) comprises 10 mg, the tertiary dose comprises 160 mg, each of the first dose fraction (F1D3) and the second fraction of the dose F2D3) comprises 80 mg, and the maximum weekly dose comprises 320 mg of the therapeutic protein.

[0089] In an exemplary embodiment of the dosing regimen, the primary dose (DI) comprises 1 mg, each of the first dose fraction (F1D1) and the second dose fraction (F2D2) of the primary dose comprising 500 mcg, the secondary dose (D2) comprises 3 mg, each of the first dose fraction (F1D2) and the second dose fraction (F2D2) of the secondary dose comprises 1.5 mg, and the maximum weekly dose comprises 3 mg of the therapeutic protein.

[0090] In an exemplary embodiment of the dosing regimen, the primary dose (DI) comprises 3 mg, each of the first dose fraction (F1D1) and the second dose fraction (F2D2) of the primary dose comprising 1.5 mg, the secondary dose (D2) comprises 9 mg, each of the first dose fraction (F1D2) and the second dose fraction (F2D2) of the secondary dose comprises 4.5 mg, and the maximum weekly dose comprises 9 mg of the therapeutic protein.

[0091] In an exemplary embodiment of the dosing regimen, the primary dose (DI) comprises 5 mg, each of the first dose fraction (F1D1) and the second dose fraction (F2D2) of the primary dose comprising 2.5 mg, the secondary dose (D2) comprises 15 mg, each of the first dose fraction (F1D2) and the second dose fraction (F2D2) of the secondary dose comprises 7.5 mg, and the maximum weekly dose comprises 15 mg of the therapeutic protein.

[0092] In an exemplary embodiment of the dosing regimen, the primary dose (DI) comprises 10 mg, each of the first dose fraction (F1D1) and the second dose fraction (F2D2) of the primary dose comprising 5 mg, the secondary dose (D2) comprises 30 mg, each of the first dose fraction (F1D2) and the second dose fraction (F2D2) of the secondary dose comprises 15 mg, and the maximum weekly dose comprises 30 mg of the therapeutic protein.

[0093] In various embodiments, the therapeutic protein is administered at a dose to maintain a serum concentration of at least about 2000 mcg / L after administration of the maximum weekly dose for the duration of the dosing regimen. In some cases, the therapeutic protein is administered at a dose to maintain a serum concentration of at least about 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3100, 3200, 3300, 3400, or 3500 mcg / L after administration of the maximum weekly dose for the duration of the dosing regimen.

[0094] In various embodiments, the therapeutic protein is administered at a dose to maintain an average serum concentration of at least about 2600 mcg / L after administration of the maximum weekly dose for the duration of the dosing regimen. In some cases, the therapeutic protein is administered at a dose to maintain an average serum concentration of at least about 2,000, 2,100, 2,200, 2,300, 2,400, 2,500, 2,700, 2,800, 2,900, 3,000, 3,100, 3,200. , 3300, 3400, 3500, 3600, 3700, 3800, 3900, or 4000 mcg / L after administration of the maximum weekly dose for the duration of the dosing regimen.

[0095] In various embodiments, the next week (Ss) of the dosing regimen is week 3 (W3), week 4 (W4), week 5 (W5), week 6 (W6), week 7 (W7), week 8 (W8), week 9 (W9), week 10 (W10), week 11 (Sil), week 12 (W12), week 13 (W13), week 14 (W14), week 15 (W15), week 16 (W16 ), week 17 (W17), week 18 (W18), week 19 (W19), week 20 (W20), week 21 (W21), week 22 (W22), week 23 (W23), week 24 (W24), week 25 (W25), week 26 (W26), week 27 (W27), week 28 (W29), week 30 (W30), week 31 (31), week 32 (32), week 33 (33), week 34 (W34) , week 35 (W35), or week 36 (W36).

[0096] In various embodiments, the second divided dose in any given week of the dosing regimen is administered within 24, 36, 48, 60, 72, 84, or 96 hours after administration of the first divided dose.

[0097] In various embodiments, the maximum weekly dose of the therapeutic protein is administered to the subject as a single dose for 1 to 8 weeks, or for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 19, 20, 21, 22, l / U I / DUO 23, 24, 25, 26 or more weeks during a weekly phase of the dosing regimen. In some cases, the maximum weekly dose of the therapeutic protein is administered to the subject as a single dose over a range of weeks that includes either the upper or lower end of the range (eg, 1-12 weeks). .

[0098] In various embodiments, the maximum weekly dose of the therapeutic protein is administered to the subject as a single dose (maintenance dose) once every two weeks for up to 2-4 weeks, or for up to 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54,55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84,85, 86, 87, 88, 89, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260 or more weeks during a biweekly, threeweekly, fourweekly, or fiveweekly phase (maintenance phase) of the dosing regimen, which may follow completion of the weekly phase of the dosing regimen (i.e., weekly dosing of the maximum weekly dose or divided dosing of primary, secondary and (optionally) tertiary doses.In some cases, the maximum weekly dose of the therapeutic protein is administered to the subject as a single dose (dose A / E / ZUZ l / U I / DUO maintenance) once every two weeks, once every three weeks, or once every four weeks during an interval of weeks that includes any of the values ​​indicated above as the upper or lower end of the interval (for example, 24-86 weeks).

[0099] In an exemplary embodiment of the dosing regimen, the primary dose comprises 1 mg, the first dose fraction (F1D1) and the second dose fraction (F2D1) each comprise 500 mcg, the secondary dose comprises 20 mg, each of the first dose fraction (F1D2) and the second dose fraction (F2D2) comprises 10 mg, the tertiary dose comprises 80 mg, each of the first dose fraction (F1D3) and the second fraction of the F2D3 dose) comprises 40 mg, and the maximum weekly dose comprises 160 mg of the therapeutic protein, where the tertiary dose is administered as a single dose (i.e., 80 mg) weekly (DS) for weeks 4 to 12 of the dosing regimen, and the maximum weekly dose is administered as a single dose (ie, 160 mg) once every two weeks (D2W) from week 14 onward of the dosing regimen.

[0100] In some instances, the dosing regimen identified above is for use in methods of treating an aggressive lymphoma (eg, mantle cell lymphoma or marginal zone lymphoma).

[0101] In an exemplary embodiment of the dosing regimen, the primary dose comprises 1 mg, the first dose fraction (F1D1) and the second dose fraction (F2D1) each comprise 500 mcg, the secondary dose comprises 20 mg, each of the first dose fraction (F1D2) and the second dose fraction (F2D2) comprises 10 mg, the tertiary dose comprises 160 mg, each of the first dose fraction (F1D3) and the second fraction of the F2D3 dose) comprises 80 mg, and the maximum weekly dose comprises 320 mg of the therapeutic protein, where the tertiary dose is administered as a single dose (i.e., 160 mg) weekly (DS) for weeks 4 to 12 of the dosing regimen, and the maximum weekly dose is administered as a single dose (ie, 320 mg) once every two weeks (D2W) from week 14 onward of the dosing regimen.

[0102] In some instances, the dosage regimen identified above is for use in methods of treating an aggressive lymphoma (eg, mantle cell lymphoma or marginal zone lymphoma). In some instances, the dosing regimen identified above is for use in methods of treating follicular lymphoma (eg, grade l-3a). In some cases, the dosing regimen identified above is for the treatment of diffuse large B-cell lymphoma (including relapsed or refractory DLBCL, eg, in patients who have failed prior CAR-T therapy). A / E / ZUZ l / U I / DUO

[0103] In various embodiments, each dose or fractionated dose of the therapeutic protein is administered to the subject over a period of 1-4, 1-5, or 1-6 hours (eg, by infusion). In some cases, the dose or fractionated dose is administered over a period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 or more hours. In some cases, each dose or fractional dose of the therapeutic protein is administered to the subject over a time interval that includes any of the values ​​indicated above as the upper or lower end of the interval (eg, 1-8 hours). In various modalities, each maximum weekly dose is administered as a single infusion.

[0104] In some embodiments of the administration regimens discussed herein, a second agent is administered as a pretreatment, or in combination with the therapeutic protein. In some cases, a spheroid, such as dexamethasone, is administered to the patient as a pretreatment prior to administration of the primary fractionated doses and secondary fractionated doses, and optionally prior to the tertiary fractionated doses. In some modalities, dexamethasone is administered to the patient approximately one to three hours prior to the first fractional dose (F1D1). In certain embodiments the dose of dexamethasone is administered by intravenous infusion. In some cases, a cytokine antagonist, 1 / U I / DUO such as an anti-IL-6 antibody or an anti-IL-6R antibody is administered in combination with the therapeutic protein during the primary, secondary, and secondary dose administrations. optionally tertiary. In some cases, a monospecific anti-CD20 antibody (eg, rituximab) is administered to the patient as a pretreatment prior to administration of the primary fractionated doses and optionally the secondary fractionated doses, and optionally prior to the tertiary fractionated doses. These pretreatment or combination administrations of the secondary agent are discontinued, in one embodiment, with single dose administration of the maximum weekly dose commencing at, for example, week 3, week 4, week 5, or week 6 of the dosage regimen so as not to artificially dampen the activity of cytokines and thereby impede the therapeutic effects of the therapeutic protein.

[0105] The terms primary dose, secondary dose, and tertiary dose, refer to the temporal sequence of administration of the antigen-binding molecule of the invention. Therefore, the primary dose is the dose that is administered at the beginning of the treatment regimen; the secondary dose is the dose that is administered after the primary dose (which includes all dose fractions of the primary dose); and tertiary doses are the doses given after the secondary dose (which includes A / E / ZUZ l / U I / DUO all fractions of the dose of the secondary dose). Primary, secondary, and tertiary doses (or dose fractions thereof) may contain an amount of the therapeutic protein as described herein. In some cases, the primary dose and the secondary dose, as well as the optional tertiary dose, may be referred to as loading doses, while subsequent maximum weekly doses may be referred to as maintenance doses. In some cases, the primary dose may be referred to as the starting dose. In some cases, the secondary dose may be referred to as the intermediate dose. In some cases, the tertiary dose may be referred to as the dose escalation.

[0106] The phrase the immediately preceding dose, in a sequence of multiple administrations, refers to the dose of the antigen-binding molecule that is administered to a patient prior to the administration of the next dose in the sequence with no intervening doses. . antigen binding molecules

[0107] Antibodies for use in connection with the present invention may be monospecific, bispecific, or multispecific. Multispecies antibodies may be specific for different epitopes of a target polypeptide or may contain antigen-binding domains specific for more than one target polypeptide. See, for example, Tutt et al., 1991, J. Immunol. 147:60-69; Kufer and others, 2004, Trends A / E / ZUZ l / U I / DUO Biotechnol. 22:238-244. The antibodies and bispecific antibodies of the present invention may be linked to or co-expressed with another functional molecule, eg, another peptide or protein. For example, an antibody or fragment thereof may be functionally linked (for example, by chemical coupling, gene fusion, non-covalent association, or in any other way) to one or more other molecular entities, such as another antibody or antibody fragment, to produce a bispecific or multispecific antibody with a second or additional binding specificity.

[0108] The present invention includes antibodies having the HCVR, LCVR and / or CDR amino acid sequences of the antibodies set forth herein, the anti-CD3 antibodies described in WO 2014 / 047231 or WO 2017 / 053856, the anti-CD20 x antiCD3 bispecific antibodies described in WO 2014 / 047231, the anti-PSMA or anti-PSMA x anti-CD3 antibodies described in WO 2017 / 023761, the anti-MUC16 or anti-MUC16 x anti-CD3 antibodies described in WO 2018 / 067331, the anti-STEAP2 or anti-STEAP2 x antiCD3 antibodies described in WO 2018 / 058001, or the anti-BCMA or anti-BCMA x anti-CD3 antibodies described in US 62 / 700,596 (filed 19 July 2018), each of which is incorporated herein by reference.

[0109] The use of the expression anti-CD3 antibody, anti-CD20 antibody, anti-PSMA antibody, antibody A / E / ZUZ I / U I / DUO anti-MUC16, anti-STEAP2 antibody, anti-BCMA antibody, or the like herein is intended to include both monospecific and bispecific antibodies that comprise the respective antigen-binding arm (for example, CD3). Therefore, the present invention includes bispecific antibodies in which one arm of an immunoglobulin binds to human CD3, and the other arm of the immunoglobulin is specific for, for example, human CD20, human PSMA, human MUGI6, human STEAP2, or human BCMA.

[0110] In certain embodiments, the CD3 binding arm binds to human CD3 and induces activation of human T cells. In certain embodiments, the CD3-binding arm weakly binds human CD3 and induces activation of human T cells. In other embodiments, the CD3 binding arm weakly binds human CD3 and induces inactivation of cells expressing the tumor associated antigen in the context of a bispecific or multispecific antibody. In other embodiments, the CD3-binding arm binds to or weakly associates with human and cynomolgus (monkey) CD3, although the binding interaction is not detectable by in vitro assays known in the art. In certain embodiments, bispecific antibodies or antigen-binding fragments for use in the present invention comprise an antigen-binding arm that binds to CD28, ICOS, HVEM, CD27, 4-1BB, Λ / C / ZUZ l / U I / DUO 0X40, DR3, GITR, CD30, SLAM, CD2, 2B4, CD226, TIM1, or TIM2 to induce T cell activation.

[0111] In certain exemplary embodiments of the present invention, the bispecific antigen-binding molecule is a bispecific antibody. Each antigen-binding domain of a bispecific antibody comprises a heavy chain variable domain (HCVR) and a light chain variable domain (LCVR). In the context of a bispecific antigen-binding molecule comprising first and second antigen-binding domains (for example, a bispecific antibody), the CDRs of the first antigen-binding domain may be designated with the prefix Al and the CDRs of the The second antigen-binding domain can be designated with the A2 prefix. Therefore, the CDRs of the first antigen-binding domain may be referred to herein as A1-HCDR1, A1-HCDR2, and A1-HCDR3; and the CDRs of the second antigen-binding domain may be referred to herein as A2-HCDR1, A2-HCDR2, and A2-HCDR3.

[0112] In one embodiment, the therapeutic protein is an anti-CD20 x anti-CD3 bispecific antibody known as REGN1979. REGN1979 comprises an anti-CD20 binding arm comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 10, an anti-CD3 binding arm comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 11, and a common light chain (corresponding to both anti-CD20 and anti-CD3 binding arms) comprising the amino acid sequence of SEQ ID NO: 12. In some cases, the mature form of the antibody may not include the C-terminal residues of the Usin of SEQ ID NO: 10 and 11. Therefore, in some cases, the anti-CD20 binding arm of REGN1979 comprises a heavy chain comprising residues 1-452 of SEQ ID NO: 10, and the antiCD3 binding arm of REGN1979 comprises a heavy chain comprising residues 1-448 of SEQ ID. NO: 11. REGN1979 also comprises the HCVR, LCVR and CDR sequences set forth in the attached sequence listing. The anti-CD20 HCVR corresponds to SEQ ID NO: 13, the anti-CD3 HCVR corresponds to SEQ ID NO: 14, and the common LCVR corresponds to SEQ ID NO: 15. The anti-CD20 domains HCDR1-HCDR2- HCDR3 correspond to SEQ ID NO: 1617-18, respectively. The anti-CD3 domains HCDR1-HCDR2HCDR3 correspond to SEQ ID NO: 19-20-21, respectively. The LCDR1-LCDR2-LCDR3 common domains correspond to SEQ ID NO: 22-23-24, respectively.

[0113] The bispecific antigen binding molecules discussed above or herein may be bispecific antibodies. In some cases, the bispecific antibody comprises a constant region of the human IgG heavy chain. In some cases, the constant region of the human IgG heavy chain is the IgG1 isotype. In some cases, the constant region of the human IgG heavy chain is the IgG4 isotype. In various embodiments, the bispecific antibody comprises a chimeric hinge that reduces Fcy receptor binding relative to a wild-type hinge of the same isotype. In some cases, the bispecific antigen-binding molecules of the present invention comprise any of the formats discussed in Brinkmann et al., MABS, 9(2) :182-212, 2017, which is incorporated herein by reference.

[0114] The first antigen-binding domain and the second antigen-binding domain may be directly or indirectly linked to each other to form a bispecific antigen-binding molecule of the present invention. Alternatively, the first antigen-binding domain and the second antigen-binding domain may each be linked to a different multimerization domain. Association of one multimerization domain with another multimerization domain facilitates association between the two antigen-binding domains, thus forming a bispecific antigen-binding molecule. A multimerization domain is any macromolecule, protein, polypeptide, peptide, or amino acid that has the ability to associate with a second multimerization domain that has the same or similar structure or constitution. For example, a multimerization domain can be a polypeptide comprising a Ch3 immunoglobulin domain. A non-limiting example of a multimerization component is an Fe portion of an immunoglobulin (comprising a Ch2-Ch3 domain), for example, an Fe domain of an IqG selected from the IgG1, IgG2, IqG3, and IgG4 isotypes, as well as any allotype within each isotype group.

[0115] The bispecific antigen-binding molecules of the present invention will typically comprise two multimerization domains, eg, two Fc domains that are each individually part of a distinct antibody heavy chain. The first and second multimerization domains can be of the same IqG isotype, eg, IqG1 / IqG1, IgG2 / IgG2, IgG4 / IgG4. Alternatively, the first and second multimerization domains may be of different IgG isotypes, eg, IgG1 / IgG2, IgG1 / IgG4, IgG2 / IgG4, etc.

[0116] In certain embodiments, the multimerization domain is an Fe fragment or an amino acid sequence from 1 to about 200 amino acids in length that contains at least one cysteine ​​residue. In other embodiments, the multimerization domain is a cysteine ​​residue, or a short cysteine-containing peptide. Other multimerization domains include peptides or polypeptides that comprise or consist of a leucine zipper, a helix loop motif, or a coiled coil motif.

[0117] Any bispecific antibody format or technology can be used to make the antigen-binding molecules of the present invention bispecific. For example, ι / u i rou or an antibody or a fragment thereof having a first antigen-binding specificity may be operatively linked (for example, by chemical coupling, gene fusion, non-covalent association, or in any other way) to one or plus different molecular entities, such as another antibody or antibody fragment having a second antigen-binding specificity to produce a bispecific antigen-binding molecule. Specific exemplary bispecific formats that may be used in the context of the present invention include, without limitation, for example, scFv or diabody-based bispecific formats, IgG-scFv fusions, Double Variable Domain (DVD)-Ig, Quadroma, buttons Knobs-lnto-holes, Common Light Chain (e.g., Common Light Chain with Buttons in Buttonholes, etc.), CrossMab, CrossFab, (SEED)body, Leucine Zipper, Duobody, IgGl / IgG2, Double Effect Fab (DAF)-IgG, and bispecific formats of Mab2{see, eg, Klein et al. 2012, mAbs 4:6, 1-11, and the references cited therein, for a review of earlier formats).

[0118] In the context of the bispecific antigen-binding molecules of the present invention, multimerization domains, eg, Fe domains, may comprise one or more amino acid changes (eg, insertions, deletions or substitutions) in comparison with the natural wild-type version of the Fe domain. For example, A / E / ZUZ I / U I / DUO The invention includes bispecific antigen-binding molecules comprising one or more modifications in the Fe domain that result in a modified Fe domain having a modified binding interaction {for example, improved or decreased) between Fe and FcRn. In one embodiment, the bispecific antigen binding molecule comprises a modification in a Ch2 or Ch3 region, wherein the modification increases the affinity of the Fe domain with respect to FcRn in an acidic environment {for example, in an endosome where the pH ranges from about 5.5 to about 6.0). Non-limiting examples of such Fe modifications include, for example, a modification at position 250 {eg, E or Q); 250 and 428 {eg, L or F) ; 252 {for example, L / Y / F / W or T) , 254 {for example, S or T) and 256 {for example, S / R / Q / E / D or T); or a modification at position 428 and / or 433 {eg, L / R / S / P / Q or K) and / or 434 {eg, H / F or Y) ; or a modification at position 250 and / or 428; or a modification at position 307 or 308 {eg, 308F, V308F), and 434. In one embodiment, the modification comprises a modification of 428L {eg, M428L) and 434S {eg, N434S); a modification of 428L, 2591 {eg V259I) and 308F {eg V308F) ; a 433K modification {eg H433K) and a 434 modification {eg 434Y); a modification of 252, 254 and 256 {for example, 252Y, 254T and 256E); a modification of 250Q and 428L {for example, T250Q and A / E / ZUZ l / U I / DUO M428L); and a modification of 307 and / or 308 (eg, 308F or 308P).

[0119] The present invention also includes bispecific antigen-binding molecules comprising a first Ch3 domain and a second Ch3 Ig domain, wherein the first and second Ch3 Ig domains differ from each other by at least one amino acid, and where where at least one amino acid difference reduces the binding of the bispecific antibody to protein A compared to a bispecific antibody that has no amino acid difference. In one embodiment, the first Ch3 Ig domain binds protein A and the second Ch3 Ig domain contains a mutation that decreases or abolishes protein A binding, such as a modification of H95R (according to IMGT numbering). of exons; H435R according to EU numbering). The second Ch3 may further comprise a modification of Y96F (according to IMGT; Y436F according to EU). See, for example, US Pat. No. 8,586,713. Other modifications that can be found within the second Ch3 include: D16E, L18M, N44S, K52N, V57M, and V82I (according to IMGT; D356E, L358M, N384S, K392N, V397M, and V422I according to EU) in the case of the IgGl antibodies; N44S, K52N and V82I (IMGT; N384S, K392N and V422I according to EU) in the case of IgG2 antibodies; and Q15R, N44S, K52N, V57M, R69K, E79Q and V82I (according to IMGT; Q355R, N384S, K392N, V397M, R409K, E419Q and V422I according to EU) in the case of IgG4 antibodies.

[0120] In certain embodiments, the Fe domain may be chimeric, combining Fe sequences derived from more than one immunoglobulin isotype. For example, a chimeric Fc domain may comprise part or all of a Ch2 sequence derived from a Ch2 region of human IgG1, human IgG2 or human IgG4, and part or all of a Ch3 sequence derived from a human IgGl, human IgG2 or human IgG4. A chimeric Fe domain may also contain a chimeric hinge region. For example, a chimeric hinge may comprise an upper hinge sequence, derived from a human IgG1, human IgG2, or human IgG4 hinge region, combined with a lower hinge sequence, derived from a human IgG1 hinge region, human IgG2 or human IgG4. A particular example of a chimeric Fc domain that can be included in any of the antigen-binding molecules set forth herein comprises, from N-terminal to C-terminal: [IgG4 ChI]-[IgG4 upper hinge]-[lower hinge of IgG2] - [IgG4 CH2] - [IgG4 CH3]. Another example of a chimeric Fc domain that can be included in any of the antigen-binding molecules set forth herein comprises, from N-terminal to C-terminal: [IgGl ChI] - [IgGl upper hinge] - [IgGl lower hinge] IgG2] - [IgG4 CH2] - [IgGl CH3]. These and other examples of chimeric Fe domains that can be included A / E / ZUZ I / U I / DUO in any of the antigen binding molecules of the present invention are described in United States Publication 2014 / 0243504, published August 28, 2014, which is incorporated herein at its entirety. Chimeric Fe domains having these general structural arrangements, and variants thereof, may have impaired binding to the Fe receptor, which in turn affects the effector function of Fe. Binding properties of antigen-binding molecules

[0121] The term binding, in the context of the binding of an antibody, immunoglobulin, antibody-binding fragment, or Fe-containing protein, to any of, for example, a predetermined antigen, such as a cell surface protein or a fragment thereof, typically refers to an interaction or association between a minimum of two molecular entities or structures, such as an antibody-antigen interaction.

[0122] For example, the binding affinity typically corresponds to a Kd value of about ΙΟ-7M or less, such as about 10~8M or less, such as about 10-9M or less, when determined, for example, by surface plasmon resonance (SPR) technology on a BIAcore 3000 instrument using the antigen as the ligand and the antibody, Ig, antibody-binding fragment, or Fe-containing protein as the A / E / ZUZ l / U I / DUO analyte (or antiligand). Cell-based binding strategies, such as fluorescence-activated cell sorting and binding (FACS) assays, are also routinely used, and FACS data correlate well with other methods, such as competitive binding of radioligands and SPR (Benedict, CA, J Immunol Methods. 1997, 201(2):223-31; Geuijen, CA, et al. J Immunol Methods. 2005, 302(1-2):68-77).

[0123] Accordingly, the antibody or antigen-binding protein of the invention binds to the predetermined antigen or cell surface molecule (receptor) that has an affinity corresponding to a Kd value that is at least ten times less than its affinity to bind to a non-specific antigen (eg, BSA, casein). In accordance with the present invention, the affinity of an antibody that corresponds to a Kd value that is equal to or ten-fold lower than that of a non-specific antigen can be considered non-detectable binding, however, this antibody can be paired with a second. antigen binding arm for the production of a bispecific antibody of the invention.

[0124] The term Kd(M) refers to the equilibrium dissociation constant of a particular antibody-antigen interaction, or the equilibrium dissociation constant of an antibody or antibody-binding fragment that binds an antigen. There is an inverse relationship between l / U I / DUO Kd and the binding affinity, therefore, the lower the Kd value, the higher the affinity, ie stronger. Therefore, the terms higher affinity or stronger affinity are related to a greater ability to form an interaction and, therefore, a lower Kd value, and, conversely, the terms lower affinity or weaker affinity are related with a lower ability to form an interaction and therefore a higher Kd value. In some circumstances, a higher binding affinity (or Kd) of a particular molecule (eg, an antibody) to its interacting partner molecule (eg, antigen X) compared to the binding affinity of the molecule (eg, an antibody) to another interacting partner molecule (for example, antigen Y) can be expressed as a binding ratio determined by dividing the highest Kd value (lower or weaker affinity) by the lower Kd value (higher or stronger affinity). ), for example, expressed as the 5-fold or 10-fold higher binding affinity, as the case may be.

[0125] The term kd (s -1 or 1 / s) refers to the dissociation rate constant of a particular antibody-antigen interaction, or the dissociation rate constant of an antibody or antibody-binding fragment. Such a value is also called a kOff value. The term ka(M-l x sec-1 or 1 / M) refers to the l / U I / DUO association rate constant of a particular antibody-antigen interaction, or the association rate constant of a binding antibody or fragment. to the antibody.

[0126] The term Ka (M-1 or 1 / M) refers to the equilibrium association constant of a particular antibody-antigen interaction, or the equilibrium association constant of an antibody or antibody-binding fragment. The equilibrium association constant is obtained by dividing the k by the kd.

[0127] The term EC50 or EC50 refers to half the maximum effective concentration, which includes the concentration of an antibody that induces a response midway between baseline and maximum after a specified exposure time. The EC50 essentially represents the concentration of an antibody where 50% of its maximum effect is observed. In certain embodiments, the EC50 value is equal to the concentration of an antibody of the invention that provides half-maximal binding to cells expressing, eg, CD3 or tumor-associated antigens (eg, CD20), as determined, for example, by a FACS binding assay. Therefore, reduced or weaker binding is observed with an EC50, or half the maximum value of the increased effective concentration.

[0128] In one embodiment, decreased binding can be defined as an increase in EC50 antibody concentration that allows binding at half-maximal amount A / E / ZUZ l / U I / DUO of the target cells.

[0129] In another embodiment, the ECso value represents the concentration of an antibody of the invention that causes semi-maximal depletion of target cells by T-cell cytotoxic activity. Thus, increased cytotoxic activity (eg, T-cell mediated inactivation of tumor cells) is observed with a decreased EC50, or half the maximum value of the effective concentration. sequence variants

[0130] The antibodies and bispecific antigen-binding molecules of the present invention may comprise one or more amino acid substitutions, insertions, and / or deletions in the framework and / or CDR regions of the heavy chain and heavy chain variable domains. slight compared to the corresponding germline sequences from which the individual antigen-binding domains were derived. Such mutations can be readily determined by comparing the amino acid sequences described herein with germline sequences available in, for example, public antibody sequence databases. The antigen-binding molecules of the present invention may comprise antigen-binding domains that are derived from any of the exemplary amino acid sequences described herein, wherein one or more amino acids are within one or more framework and / or CDR regions. mutate to the corresponding residues of the germline sequence from which the antibody is derived, or to corresponding residues of another human germline sequence, or to a conservative amino acid substitution of the corresponding germline residues (such changes sequence mutations are collectively referred to herein as germline mutations). A person skilled in the art can readily produce, from the heavy and light chain variable region sequences described herein, numerous antibodies and antigen-binding fragments comprising one or more individual germline or combinations thereof. In certain embodiments, all framework and / or CDR residues within the Vh and / or Vl domains are mutated back to residues found in the original germline sequence from which the antigen-binding domain was originally derived. In other embodiments, only certain residues are mutated back to the original germline sequence, for example, only mutated residues found within the first 8 amino acids of FR1 or within the last 8 amino acids of FR4, or only mutated residues found within the first 8 amino acids of FR4. within CDR1, CDR2 or CDR3. In other embodiments, one or more of the framework and / or CDR residues are mutated to the corresponding residues of a sequence in the line. A / E / ZUZ I / U I / DUO different germline (ie, a germline sequence that is different from the germline sequence from which the antigen-binding domain is originally derived). In addition, antigen-binding domains may contain any combination of two or more germline mutations within the framework and / or CDR regions, for example, where certain individual residues are mutated to the corresponding residue of a germline sequence. particular, while residues other than the original germline sequence are maintained or mutated to the corresponding residue of a different germline sequence. Once obtained, antigen-binding domains containing one or more germline mutations can be readily analyzed for one or more desired properties, such as improved binding specificity, increased binding affinity, improved agonist or antagonist biological properties. (as the case may be), reduced immunogenicity, etc. Bispecific antigen-binding molecules comprising one or more antigen-binding domains obtained in this general manner are included within the present invention. pH dependent binding

[0131] The present invention includes antibodies and bispecific antigen-binding molecules with pH-dependent binding characteristics. For example, an antibody of the present invention may exhibit reduced binding to, for example, a tumor antigen such as CD20 at acidic pH compared to neutral pH. Alternatively, the antibodies of the invention may exhibit enhanced binding to, for example, a tumor antigen such as CD20 at acidic pH compared to neutral pH. The term "acidic pH" includes pH values ​​less than about 6.2, for example, about 6.0, 5.95, 5.9, 5.85, 5.8, 5.75, 5.7, 5.65, 5.6, 5.55, 5.5, 5.45, 5.4, 5.35, 5.3, 5.25, 5.2, 5.15, 5.1, 5.05, 5.0 or less. By "neutral pH" is meant a pH of about 7.0 to about 7.4. The term "neutral pH" includes pH values ​​of approximately 7.0, 7.05, 7.1, 7.15, 7.2, 7.25, 7.3, 7.35, and 7.4.

[0132] In certain instances, reduced binding...at acidic pH compared to neutral pH is expressed in terms of a ratio of the Kd value of binding of the antibody to its antigen at acidic pH to the Kd value of binding of the antibody to its antigen at neutral pH (or vice versa). For example, an antibody, or antigen-binding fragment thereof, may be considered to exhibit reduced binding to, for example, CD20 at acidic pH compared to neutral pH for purposes of the present invention if the antibody or antigen-binding fragment thereof antigen binding thereof exhibits an acid / neutral Kd ratio of approximately 3.0 or greater. In certain exemplary l / U I / DUO embodiments, the acidic / neutral Kd ratio for an antibody or antigen-binding fragment of the present invention can be approximately 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 20.0. 25.0, 30.0, 40.0, 50.0, 60.0, 70.0, 100.0 or greater.

[0133] Antibodies with pH-dependent binding characteristics can be obtained, for example, by screening a population of antibodies for reduced (or enhanced) binding to a particular antigen at acidic pH, compared to neutral pH. Furthermore, modifications of the antigen-binding domain at the amino acid level can produce antibodies with pH-dependent characteristics. For example, by replacing one or more amino acids of an antigen-binding domain (eg, within a CDR) with a histidine residue, an antibody with reduced antigen-binding at acidic pH relative to pH can be obtained. neutral. Antibodies comprising Fe variants

[0134] According to certain embodiments of the present invention, there are provided antibodies and bispecific antigen-binding molecules comprising an Fc domain comprising one or more mutations that increase or decrease antibody binding to the FcRn receptor, for example , at acidic pH compared to neutral pH. By For example, the present invention includes antibodies comprising a mutation in the Ch2 or Ch3 region of the Fe domain, wherein the mutation increases the affinity of the Fe domain to FcRn in an acidic environment (for example, in an endosome where the pH varies from about 5.5 to about 6.0). Such mutations can result in an increased half-life of the antibody in the serum when administered to an animal. Non-limiting examples of such Fe modifications include, for example, a modification at position 250 (eg, E or Q); 250 and 428 (eg Lo F); 252 (eg, L / Y / F / W or T), 254 (eg, S or T), and 256 (eg, S / R / Q / E / D or T); or a modification at position 428 and / or 433 (eg H / L / R / S / P / Q or K) and / or 434 (eg H / F or Y); or a modification at position 250 and / or 428; or a modification at position 307 or 308 (eg, 308F, V308F), and 434. In one embodiment, the modification comprises a modification of 428L (eg, M428L) and 4343 (eg, N434S); a modification of 428L, 2591 (eg V259I) and 308F (eg V308F); a 433K modification (eg, H433K) and a 434 modification (eg, 434Y); a modification of 252, 254 and 256 (for example, 252Y, 254T and 256E); a modification of 250Q and 428L (for example, T250Q and M428L); and a modification of 307 and / or 308 (eg, 308F or 308P).

[0135] A / E / ZUZ l / U I / DUO For example, the present invention includes antibodies and bispecific antigen-binding molecules that comprise an Fc domain comprising one or more pairs or groups of mutations selected from the group consisting of: 250Q and 248L {eg, T250Q and M248L); 252Y, 254T and 256E {for example, M252Y, S254T and T256E) ; 428L and 434S {for example, M428L and N434S) ; and 433K and 434F {eg, H433K and N434F). All possible combinations of the above Fe domain mutations, and other mutations within the antibody variable domains described herein, are contemplated within the scope of the present invention. Preparation of antigen-binding domains and construction of bispecific molecules

[0136] Antigen binding domains specific for particular antigens can be prepared by any antibody generating technology known in the art. Once obtained, two different antigen-binding domains specific for two different antigens (for example, CD3 and CD20), can be suitably arranged with respect to one another to produce a bispecific antigen-binding molecule of the present invention by using methods of routine. (A discussion of exemplary bispecific antibody formats that can be used to construct the bispecific antigen-binding molecules of the present invention is provided elsewhere herein.) In certain embodiments, one or more of the individual components (eg, the heavy and light chains) of the multispecific antigen-binding molecules of the invention are derived from chimeric, humanized, or fully human antibodies. Methods for producing such antibodies are well known in the art. For example, one or more of the heavy and / or light chains of the bispecific antigen-binding molecules of the present invention can be prepared using the VELOCIMMUNE™ technology. Using VELOCIMMUNE™ technology (or any other human antibody-generating technology), high-affinity chimeric antibodies to a particular antigen (eg, CD3 or CD20) that have a human variable region and a constant region are initially isolated. of mouse. Antibodies are characterized and selected based on desirable characteristics, including affinity, selectivity, epitope, etc. The mouse constant regions are replaced with a desired human constant region to generate fully human heavy and / or light chains that can be incorporated into the bispecific antigen-binding molecules of the present invention.

[0137] Genetically modified animals can be used to make human bispecific antigen-binding molecules. For example, a genetically engineered mouse that is incapable of rearranging and expressing an endogenous mouse immunoglobulin light chain variable sequence can be used, wherein the mouse expresses only one or two human light chain variable domains encoded by the human immunoglobulin sequences operably linked to the mouse constant kappa gene at the endogenous mouse kappa locus. Such genetically modified mice can be used to produce fully human bispecific antigen-binding molecules comprising two different heavy chains that are associated with an identical light chain comprising a variable domain derived from one of two variable chain region gene segments. different human light. (See, for example, US 2011 / 0195454). Fully human refers to an antibody, or antigen-binding fragment or immunoglobulin domain thereof, comprising an amino acid sequence encoded by a DNA derived from a human sequence over the full length of each antibody polypeptide or fully human-binding fragment. antigen or immunoglobulin domain thereof. In some cases, the fully human sequence is derived from a protein endogenous to a human. In other cases, the fully human protein or protein sequence comprises a chimeric sequence where each component sequence is derived from a human sequence. Although not limited by any theory, chimeric proteins or chimeric sequences are generally designed to minimize the creation of immunogenic epitopes at junctions of component sequences, for example, compared to l / U I / DUO any region or domain. of wild-type human immunoglobulin. bioequivalent

[0138] The present invention encompasses antigen-binding molecules that have amino acid sequences that vary from the exemplary molecules described herein, but that retain the ability to bind to the same antigen(s). Such variant molecules may comprise one or more amino acid additions, deletions, or substitutions when compared to the parent sequence, but exhibit biological activity that is essentially equivalent to that of the disclosed bispecific antibodies or antigen-binding molecules. The present invention includes antigen binding molecules that are bioequivalent to any of the exemplary antigen binding molecules set forth herein. Two antigen-binding proteins, or antibodies, are considered bioequivalent if, for example, they are pharmaceutical equivalents or pharmaceutical alternatives whose rate and degree of absorption do not show a significant difference when administered at the same molar dose under similar experimental conditions, either in single or multiple doses. Some antigen-binding proteins will be considered equivalent or pharmaceutical alternatives if they are equivalent in the extent of their absorption but not in the rate of absorption l / U I / DUO, and yet can be considered bioequivalent because such differences in rate absorption rates are intentional and are reflected in the labeling, are not essential to achieve effective concentrations of the drug in the body, eg, in chronic use, and are considered medically insignificant for the particular drug under study.

[0139] In one embodiment, two antigen-binding proteins are bioequivalent if there are no clinically significant differences in their safety, purity, and potency.

[0140] In one embodiment, two antigen-binding proteins are bioequivalent if a patient can switch one or more times between the reference product and the biologic without an expected increased risk of adverse effects, including a clinically significant change in immunogenicity, or decreased effectiveness, compared to continued therapy without such a change.

[0141] In one embodiment, two antigen-binding proteins are bioequivalent if they both act through a common mechanism(s) of action for the condition(s) of use, to the extent such mechanisms are known.

[0142] Bioequivalence can be demonstrated by both in vivo and in vitro methods. Bioequivalence measures include, for example, (a) an in vivo test in humans or A / E / ZUZ l / U I / DUO other mammals, where the concentration of the antibody or its metabolites is measured in blood, plasma, serum or other biological fluid as a function of time; (b) an in vitro test that correlates with and is reasonably predictive of human in vivo bioavailability data; (c) an in vivo test in humans or other mammals where the appropriate acute pharmacological effect of the antibody (or its target) is measured as a function of time; and (d) in a well-controlled clinical trial that establishes the safety, efficacy, or bioavailability or bioequivalence of an antigen-binding protein.

[0143] Bioequivalent variants of the exemplary antibodies and bispecific antigen-binding molecules set forth herein can be constructed by, for example, various residue or sequence substitutions or by deletion of terminal or internal sequences or residues gue they are not required for biological activity. For example, cysteine ​​residues that are not essential for biological activity can be removed or replaced with other amino acids to prevent unnecessary or incorrect intramolecular disulfide bond formation after renaturation. In other contexts, bioequivalent antigen-binding proteins may include variants of the exemplary antibodies and the bispecific antigen-binding molecules discussed herein that comprise amino acid changes that modify the A / E / ZUZ l / U I / DUO glycosylation characteristics of molecules, for example, mutations that abolish or remove glycosylation. Therapeutic formulation and administration

[0144] The present invention provides pharmaceutical compositions comprising the antigen binding molecules of the present invention. The pharmaceutical compositions of the invention are formulated with suitable carriers, excipients, and other agents that provide improved transfer, delivery, tolerability, and the like. A multitude of suitable formulations can be found in the form known to all pharmaceutical chemists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid-containing vesicles (cationic or anionic) (such as LIPOFECTIN™, Life Technologies, Carlsbad, CA), DNA conjugates, anhydrous absorption, oil-in-water and water-in-oil emulsions, carbowax emulsions (polyethylene glycols of various molecular weights), semisolid gels and semisolid mixtures containing carbowax. See also Powell et al. Compendium of excipients for parenteral formulations PDA (1998) J Pharm Sci Technol 52:238311.

[0145] Several delivery systems are known that can be used to deliver the pharmaceutical composition of l / U I / DUO the invention, for example, encapsulation in liposomes, microparticles, microcapsules, recombinant cells that can express the mutant viruses, endocytosis mediated by receptors (see, eg, Wu et al., 1987, J. Biol. Chem. 262:4429-4432). Methods of introduction include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The composition can be administered by any convenient route, for example, by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (for example, oral mucosa, rectal and intestinal mucosa, etc.), and can be Administer together with other biologically active agents. Administration can be systemic or local. In one embodiment, the therapeutic proteins of the invention are administered by intravenous infusion.

[0146] A pharmaceutical composition of the present invention can be administered subcutaneously or intravenously with a standard needle and syringe. Furthermore, with respect to subcutaneous administration, a pen-type administration device has applications in the administration of a pharmaceutical composition of the present invention. Said pen-like delivery device may be reusable or disposable. A reusable pen-type administration device typically uses A / E / ZUZ l / U I / DUO a replaceable cartridge containing a pharmaceutical composition. Once all of the pharmaceutical composition has been dispensed into the cartridge and the cartridge is empty, the cartridge can be easily discarded and replaced with a fresh cartridge containing the pharmaceutical composition. The pen-type delivery device can then be reused. In a disposable pen-type administration device, the cartridge is not replaced. Instead, the disposable pen-like delivery device comes pre-filled with the pharmaceutical composition that is held in a reservoir within the device. Once the reservoir is emptied of the pharmaceutical composition, the entire device is discarded.

[0147] Numerous pen-type delivery devices and reusable autoinjectors have applications in the subcutaneous administration of a pharmaceutical composition of the present invention. Examples include, but are not limited to, AUTOPEN™ (Owen Mumford, Inc., Woodstock, RN), DISETRONIC™ Pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75 / 25™ Pen, HUMALOG™ Pen, Ballpoint Pen HUMALIN 70 / 30™ (Eli Lilly and Co., Indianapolis, IN), NOVOPEN™ I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR™ (Novo Nordisk, Copenhagen, Denmark), BD™ Ballpoint Pen (Becton Dickinson, Franklin Lakes, NJ) , OPTIPEN™, OPTIPEN PRO™, OPTIPEN STARLET™, and OPTICLIK™ (sanofi-aventis, Frankfurt, Germany), to name a few. Examples of l / U I / DUO disposable pen-type administration devices that have applications in the subcutaneous administration of a pharmaceutical composition of the present invention include, but are not limited to, the SOLOSTAR™ pen (sanofi-aventis), the FLEXPEN™ ( Novo Nordisk), and the KWIKPEN™ (Eli Lilly), the SURECLICK™ autoinjector (Amgen, Thousand Oaks, CA), the PENLET™ (Haselmeier, Stuttgart, Germany), the EPIPEN (Dey, L.P.), and the HUMIRA™ pen (Abbott Labs, Abbott Park IL), to name a few.

[0148] In certain situations, the pharmaceutical composition can be administered via a controlled release system. In one embodiment, a pump may be used (see Langer, supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng. 14:201). In another embodiment, polymeric materials can be used; see, Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Pres., Boca Raton, Florida. In yet another embodiment, a controlled release system can be placed in close proximity to the target of the composition, requiring only a fraction of the systemic dose (see, for example, Goodson, 1984, in Medical Applications of Controlled Release, supra , vol. 2, pp. 115-138). Other controlled release systems are discussed in the review by Langer, 1990, Science 249:1527-1533.

[0149] Injectable preparations may include dosage forms for intravenous, subcutaneous, intracutaneous and intramuscular injections, drip infusions, etc. These injectable preparations can be prepared by publicly known methods. For example, injectable preparations can be prepared, for example, by dissolving, suspending or emulsifying the antibody, or its salts described above, in a sterile aqueous medium or an oily medium conventionally used for injections. As the aqueous medium for injections, there is, for example, a physiological saline solution, an isotonic solution containing glucose and other auxiliary agents, etc., which can be used in combination with a suitable solubilizing agent, such as an alcohol (for example, ethanol), a polyalcohol (eg, propylene glycol, polyethylene glycol), a nonionic surfactant [eg, polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)], etc. As the oily medium, for example, sesame oil, soybean oil, etc., which can be used in combination with a solubilizing agent, such as benzyl benzoate, benzyl alcohol, etc., are used. The injection thus prepared is preferably filled into a suitable ampoule. Therapeutic uses of antigen-binding molecules

[0150] The present invention includes methods comprising administering to a subject in need thereof a therapeutic composition comprising a therapeutic protein. The therapeutic composition can comprise any of the antibodies or bispecific antigen binding molecules described herein and a pharmaceutically acceptable carrier or diluent. The expression "a subject in need thereof" means a human or non-human animal exhibiting one or more symptoms or indications of cancer (for example, a subject expressing a tumor or suffering from any of the types of cancer mentioned herein below). ).

[0151] The antibodies and bispecific antigen-binding molecules of the invention (and therapeutic compositions comprising them) are useful, inter alia, for the treatment of any disease or disorder where stimulation, activation and / or targeting of an immune response would be beneficial. In particular, the antibodies or bispecific antigen-binding molecules of the present invention can be used for the treatment, prevention and / or amelioration of any disease or disorder associated with or mediated by, for example, the expression or activity of CD20, PMSA, MUC16, STEAP2, or BCMA or proliferation of CD20+, PSMA+, MUC16+, STEAP2+, or BCMA+ cells. The mechanism of action by which the therapeutic methods of the invention are achieved includes inactivation of cells expressing said antigens in the presence of effector cells, for example, by CDC, apoptosis, ADCC, phagocytosis, or by a combination of two. or more of these mechanisms.

[0152] The antigen-binding molecules of the present invention can be used to treat, for example, primary and / or metastatic tumors arising in the brain and A / E / ZUZ l / U I / DUO the meninges, head and neck, oropharynx, lung and bronchial tree, gastrointestinal tract, male and female reproductive tract, muscle, bone, skin and appendages, connective tissue, spleen, immune system, cells blood-forming and bone marrow, liver and urinary tract, kidney, bladder, and / or special sensory organs such as the eye. In certain embodiments, the bispecific antibodies and antigen-binding molecules of the invention are used to treat one or more of, but are not limited to, the following cancers: pancreatic carcinoma, head and neck cancer, prostate cancer, gliomas malignant cancers, osteosarcoma, colorectal cancer, gastric cancer (eg, MET-amplified gastric cancer), malignant mesothelioma, multiple myeloma, ovarian cancer, small cell lung cancer, non-small cell lung cancer, synovial sarcoma, cancer of the thyroid, breast cancer, melanomaglioma, breast cancer (for example, ductal or intraductal breast carcinoma, squamous cell carcinoma, esophageal cancer, clear cell renal cell carcinoma, chromophobe renal cell carcinoma, oncocytoma (renal), cell carcinoma transitional (renal) carcinoma, urothelial carcinoma, adenocarcinoma (bladder), or small cell carcinoma (bladder) According to certain embodiments of the present invention, antibodies or bispecific antibodies are useful for treating a patient afflicted with a refractory or resistant to treatment for cancer A / E / ZUZ l / U I / DUO example, castration resistant prostate cancer.

[0153] In some embodiments, the antigen-binding molecule is an anti-CD3 x anti-CD20 bispecific antibody useful for treating a cancer that expresses CD20, including non-Hodgkin's lymphoma, Hodgkin's lymphoma, chronic lymphocytic leukemia, acute lymphoblastic leukemia, small lymphocytic lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, marginal zone lymphoma, Waldenstrom macroglobulinemia, primary mediastinal B-cell lymphoma, lymphoblastic lymphoma, or Burkitt's lymphoma. In some embodiments, the cancer is follicular lymphoma. In some forms, the cancer is diffuse large B-cell lymphoma (DLBCL). In some forms, the cancer is mantle cell lymphoma. In some modalities, the cancer is marginal zone lymphoma. In some modalities, the cancer is follicular lymphoma and the maximum weekly dose of the bispecific antibody is 80 mg. In some modalities, the cancer is DLBCL and the maximum weekly dose of the bispecific antibody is 80 mg. In some modalities, the cancer is DLBCL and the maximum weekly dose of the bispecific antibody is 160 mg. In some modalities, the cancer is DLBCL and the maximum weekly dose of the bispecific antibody is 320 mg. In any of these modalities, or others discussed in the present description, the cancer patient A / E / ZUZ l / U I / DUO may have been pretreated with anti-CD20 monospecific antibody therapy.

[0154] Non-Hodgkin's lymphoma (NHL) is the most common hematologic malignancy. Within a heterogeneous group of NHL, 85-90% are of B-cell origin and include follicular lymphoma (LE), diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL) and various other B-NHL. Anti-CD20 antibodies in combination with chemotherapy are the standard of care for the treatment of B-NHL; however, despite initial responses, many patients relapse, often with progressively shorter durations of response in later lines of therapy and with poor outcomes. Thus, in some embodiments, the antigen-binding molecule is a bispecific anti-CD3 x anti-CD20 that binds to CD3+ T cells and CD20+ B cells, targeting CD20+ tumor cells through mediated cytotoxicity. by T cells. In some instances, the anti-CD3 x CD20 bispecific antibody is for the treatment of a B-cell cancer (eg, NHL) in a subject who has failed prior therapy with an anti-CD20 monospecific antibody .

[0155] For patients with less than a complete response to CAR-T therapy, outcomes are generally poor and there are no standard therapeutic options. Therefore, in some cases, the anti-CD3 x CD20 bispecific antibody of the present invention is for the treatment of a B-cell cancer (for example, an NHL such as DLBCL) in a subject who has failed prior therapy. CAR-T or not responding to prior CAR-T therapy (eg, anti-CD19 CAR-T therapy).

[0156] In some embodiments, the antigen binding molecule is an anti-CD3 x anti-PSMA bispecific antibody useful for treating a PSMA-expressing cancer including prostate cancer, kidney cancer, bladder cancer, liver cancer, colorectal cancer and gastric cancer. In some embodiments, the cancer is prostate cancer (eg, castration resistant prostate cancer).

[0157] In some embodiments, the antigen binding molecule is an anti-CD3 x anti-MUC16 bispecific antibody useful for treating a cancer that expresses MUC16, including ovarian cancer, breast cancer, pancreatic cancer, cancer non-small cell lung cancer, mass-forming intrahepatic cholangiocarcinoma, adenocarcinoma of the cervix, and adenocarcinoma of the gastric tract. In some embodiments, the cancer is ovarian cancer.

[0158] In some embodiments, the antigen-binding molecule is an anti-CD3 x anti-STEAP2 bispecific antibody useful for treating a cancer expressing STEAP2 including prostate cancer, bladder cancer, neck cancer ι / u i rouo uterine cancer, lung cancer, colon cancer, kidney cancer, breast cancer, pancreatic cancer, stomach cancer, uterine cancer, and ovarian cancer. In some embodiments, the cancer is prostate cancer {eg, castration resistant prostate cancer).

[0159] In some embodiments, the antigen binding molecule is an anti-CD3 x anti-BCMA bispecific antibody useful for treating a BCMA-expressing cancer including multiple myeloma or other B-cell or plasma cell cancers, such as multiple myeloma. Waldenstrom's macroglobulinemia, Burkitt's lymphoma, and diffuse large B-cell lymphoma, non-Hodgkin's lymphoma, chronic lymphocytic leukemia, follicular lymphoma, mantle cell lymphoma, marginal zone lymphoma, lymphoplasmacytic lymphoma , and Hodgkin lymphoma. In some forms, the cancer is multiple myeloma. Combination therapies

[0160] The present invention provides methods comprising administration of a pharmaceutical composition comprising any of the exemplary antibodies and bispecific antigen binding molecules described herein in combination with one or more additional therapeutic agents. Exemplary additional therapeutic agents that may be combined with or administered in combination with an antigen binding molecule of the A / E / ZUZ I / U I / DUO present invention include, for example, an antitumor agent (for example, chemotherapeutic agents including melphalan, vincristine (Oncovin), cyclophosphamide (Cytoxan), etoposide (VP-16), doxorubicin (Adriamycin ), liposomal doxorubicin (Doxil), obendamustine (Treanda), or any other known to be effective in treating a plasma cell tumor in a subject). In certain embodiments, the second therapeutic agent is a regimen comprising radiation therapy or hematopoietic stem cell transplantation. In certain embodiments, the second therapeutic agent can be an immunomodulatory agent. In certain embodiments, the second therapeutic agent may be a proteasome inhibitor, including bortezomib (Velcade), carfilzomib (Kyprolis), ixazomib (Ninlaro). In certain embodiments, the second therapeutic agent may be a histone deacetylase inhibitor such as panobinostat (Farydak). In certain embodiments, the second therapeutic agent can be a monoclonal antibody, an antibody-drug conjugate, a bispecific antibody conjugated to an antitumor agent, an immune checkpoint inhibitor, or combinations thereof. The pharmaceutical compositions of the present invention (for example, pharmaceutical compositions comprising a bispecific antigen-binding molecule as described herein) may also be administered as part of a therapeutic I / U I / DUO regimen comprising one or more selected therapeutic combinations of a monoclonal antibody other than those described herein, which can interact with a different antigen on the surface of the plasma cell, a bispecific antibody, which has an arm that binds to an antigen on the surface of the plasma cell tumor cell and the other arm binds to an antigen on the T cell, an antibody-drug conjugate, a bispecific antibody conjugated to an antitumor agent, a checkpoint inhibitor, for example, one that targets PD-1 or CTLA -4, or their combinations. In certain embodiments, checkpoint inhibitors may be selected from PD-1 inhibitors, such as pembrolizumab (Keytruda), nivolumab (Opdivo), or cemiplimab. In certain embodiments, the checkpoint inhibitors may be selected from PD-L1 inhibitors, such as atezolizumab (Tecentriq), avelumab (Bavencio), or Durvalumab (Imfinzi)). In certain embodiments, checkpoint inhibitors may be selected from CTLA-4 inhibitors, such as ipilimumab (Yervoy). Other combinations that can be used in conjunction with an antibody of the invention are described above.

[0161] The present invention also includes therapeutic combinations comprising any of the antigen binding molecules mentioned herein and an inhibitor of one or more of VEGF, Ang2, DLL4, EGFR, ErbB2, ErbB3, A / E / ZUZ l / U I / DUO 100 ErbB4, EGFRvIII, cMet, IGF1R, B-raf, PDGFR-a, PDGFR-β, FOLH1 (PSMA), PRLR, STEAP1, STEAP2, TMPRSS2, MSLN, CA9, uroplakin, where the inhibitor is an aptamer, an antisense molecule , a ribozyme, siRNA, peptibody, nanobody or antibody fragment (for example, Fab fragment; F(ab')2 fragment, Fd fragment; Fv fragment; scFv; dAb fragment; or other genetically modified molecules, such as diabodies, tribodies, tetrabodies, minibodies and minimal recognition units). The antigen binding molecules of the invention may also be administered and / or co-formulated in combination with antivirals, antibiotics, analgesics, and / or ΑΙΝΕ. The antigen binding molecules of the invention may also be administered as part of a treatment regimen that also includes radiation treatment and / or conventional chemotherapy.

[0162] The therapeutically active additional component may be administered just before, simultaneously with, or shortly after administration of an antigen binding molecule of the present invention; (For the purposes of the present description, such administration regimens are considered the administration of an antigen-binding molecule in combination with an additional therapeutically active component).

[0163] The present invention includes pharmaceutical compositions in which an antigen-binding molecule of the A / E / ZUZ l / U I / DUO 101 The present invention is co-formulated with one or more therapeutically active additional ingredients, as described elsewhere herein. EXAMPLES

[0164] The following examples are set forth to provide those skilled in the art with a complete disclosure and description of how to make and use the methods and compositions of the invention, and are not intended to limit the scope of what the inventors consider to be their invention. Efforts have been made to ensure accuracy with respect to the numbers used (eg, amounts, temperature, etc.), but some experimental error and bias should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric. Example 1: Clinical evaluation and dose escalation of a bispecific antibody

[0165] The clinical study described below is a phase 1 multicenter open-label study to investigate the safety and tolerability of REGN1979, an anti-CD20 x anti-CD3 bispecific monoclonal antibody, in patients with previously treated CD20+ B-cell malignancies. with antibody therapy directed at CD20. Λ / C / ZUZ l / U I / DUO 102

[0166] Objectives: The primary objectives of the study were to assess the safety, tolerability, and dose-limiting toxicities (DLT) of REGN1979 administered intravenously (IV) and to study the antitumor activity of REGN1979 in expansion cohorts comprising the diffuse large B-cell lymphoma (DLBCL) after failure of chimeric antigen receptor T-cell therapy (CART), aggressive lymphoma (other than DLBCL after failure of CAR-T therapy), follicular lymphoma (FLE) ) grade 13a, and chronic lymphocytic leukemia (CLL). The secondary objectives of the study were: (1) to characterize the pharmacokinetic (PK) profile of REGN1979; (2) to assess the immunogenicity of REGN1979; (3) to study the preliminary antitumor activity of REGN1979 administered to patients with CD20+ B-cell malignancies (non-Hodgkin's lymphoma [NHL] previously treated with anti-CD20 antibody therapy, or chronic lymphocytic leukemia [CLL]); and (4) to study the preliminary antitumor activity of REGN1979 in the dose escalation portion of the study. Minimal residual disease (MRD) assessments were performed in the CLL patients. The exploratory objectives of the study were to evaluate biomarkers that can be correlated with the mechanism of action, observed toxicity, and potential antitumor activity including, but not limited to: (1) cytokine profiling and assessment of inflammatory markers (for A / C / ZUZ l / U I / DUO 103 example, C-reactive protein [CRP]); (2) subsets of peripheral blood B cells and T cells and immunological phenotyping; and (3) changes in gene expression in peripheral blood.

[0167] Study Design: Patients were assigned to a dose level (ND) cohort consisting of an initial starting dose, followed by higher escalating doses for the second administration and subsequent doses. Patients were enrolled based on indication (NHL or CLL). In each ND, there were 2 cohorts (one for each indication), with 3-6 patients per NHL cohort and 1-6 patients per CLL cohort. Patients with small lymphocytic lymphoma (SLL) were enrolled in the CLL group and followed for NHL evaluations.

[0168] Patients who initially showed clinical benefit and who subsequently relapsed or progressed or had a suboptimal response to treatment may have been re-treated with REGN1979 at the highest ND considered tolerable at the time of relapse or progression .

[0169] Patients underwent screening procedures for eligibility within 28 days prior to initial REGN1979 administration. Patients were enrolled sequentially based on indication (NHL or CLL) in order of sponsor eligibility confirmation until each cohort was completed as per A / E / ZUZ l / U I / DUO 104 the criteria of the protocol.

[0170] There were independent dose escalation cohorts for NHL and CLL at each ND. Each ND consisted of an initial dose and a second and subsequent dose, which was greater than the starting dose, as long as the initial dose was tolerated.

[0171] The dose escalation followed a traditional 3+3 dose escalation design for NHL patients. Three to six patients per cohort were planned depending on the observed toxicity.

[0172] Dose escalation followed a modified 3+3 with an accelerated titration component for CLL patients. Depending on the observed toxicity, 1 to 6 patients per cohort were planned.

[0173] Following completion of the dose escalation phase and determination of the recommended dose for further studies in NHL patients, three expansion cohorts were opened for 1) DLBCL patients after therapy failure with CAR-T (20 patients), 2) aggressive lymphoma (other than DLBCL after failure of CAR-T therapy) (40 patients; among whom 20 patients were each enrolled in aggressive lymphoma cohort 1 [dose 160 mg dose escalation]), and Cohort 2 [80 mg dose escalation]), and 3) patients with relapsed / refractory grade l-3a follicular lymphoma (60 patients; among whom 30 were each enrolled in the cohort 1 [step dose of 80 A / E / ZUZ l / U I / DUO 105 mg] and cohort 2 [160 mg dose escalation]). The escalating weekly dose of REGN1979 for each of these expansion cohorts (except for aggressive lymphoma cohort 2 and follicular lymphoma cohort 1) was established at 160 mg weekly, followed by maintenance treatment every 2 weeks with 320 mg. by REGN1979. In aggressive lymphoma cohort 2 and follicular lymphoma cohort 1, the weekly dose escalation of REGN1979 was 80 mg, and the every 2-week maintenance dose of REGN1979 was 160 mg. For patients assigned to an 80 mg dose escalation, patients received 80 mg weekly over a 4-week lead-in period, after initial dose escalation, followed by an additional 8 weekly doses, and treatment with 160 mg every 2 weeks until progression, with an option to discontinue treatment thereafter that the patient has shown a durable response for at least 9 months after the initial demonstration of a RG. For patients assigned to a 160 mg dose escalator, patients received 160 mg weekly for a 4-week lead-in period, after the initial dose escalation, followed by an additional 8 weekly doses, and treatment with 320 mg every 2 weeks until progression, with an option to discontinue treatment after the patient has shown a durable response for at least 9 months after the A / E / ZUZ l / U I / DUO initial demonstration of a CR. 106

[0174] The utility of a starting dose of rituximab was studied prior to the first administration of REGN1979 in patients with NHL to determine whether this intervention can decrease the incidence and severity of infusion-related reaction (IRR) and syndrome. cytokine release (SLC). In the initial rituximab cohort, REGN1979 was administered using a 160 mg entry dose during weekly treatment and 320 mg during every 2-week maintenance treatment. Subsequently, another 24 patients were to be treated with the optimal dose and dosage regimen; Together with the 6 patients in the initial rituximab group treated with the optimal dose, a total of 30 patients were reviewed for safety and tolerability.

[0175] In the first ND for the dose escalation portion, there was a required 48 hour waiting period between initial study drug administrations for the first 3 patients within the same indication. Subsequent patients in the first ND were not treated on the same day, regardless of the indication. In subsequent cohorts, provided no unexpected toxicity was observed in earlier or within-cohort cohorts, initial infusions for the first 3 patients were administered at least 24 hours apart.

[0176] After each patient cohort was enrolled, treated, and the observation period of 107 NDT, the subsequent ND cohorts were determined to open for enrollment (or the current open ND cohort expansion) was determined once both the sponsor and investigator(s) had reviewed the safety data.

[0177] The NDT observation period was defined as the first 28 days of treatment, which in this study corresponds to the lead-in period. During induction, patients were treated with 4 weekly administrations of REGN1979.

[0178] To be assessed as NDT, an individual patient must have received at least the first 2 administrations of REGN1979 (week 1 day 1 [initial or primary dose] and week 2 day 1 [secondary and subsequent dose]), or experienced a NDT. Patients enrolled at ND11 and above must have received at least the first 3 administrations of REGN1979 (initial dose week 1, secondary or intermediate dose week 2, and tertiary or upper dose escalation week 3) or experienced an NDT. In addition, the patient must have been evaluated for at least 28 days from the first administration, and at least 21 days from the second administration.

[0179] Dose escalation and cohorts for NHL and CLL patients are shown in Table 1, below. l / U I / DUO Table 1. Dose escalation and cohorts. 108 Dose Level Starting Dose (mcg) Intermediate Dose (mcg) Maximum Dose (Dose Escalation) (mcg) NHL n CLL n ND1 30 - 100 Cohort 1 3-6 Cohort 1 1-6 ND2 100 - 300 Cohort 2 3-6 Cohort 2 1-6 ND3 300 - 1000 Cohort 3 3-6 Cohort 3 1-6 ND4 1000 - 2000 Cohort 4 3-6 Cohort 4 1-6 ND5 1000 - 3000 Cohort 5 3-6 Cohort 5 1-6 ND6 1000 - 4000 Cohort 6 3-6 Cohort 6 1-6 ND7 1000 - 5000 Cohort 7 3-6 Cohort 7 1-6 ND8 1000 - 6000 Cohort 8 3-6 Cohort 8 1-6 ND9 1000 - 7000 Cohort 9 3-6 Cohort 9 1 -6 ND10 1000 - 8000 Cohort 10 3-6 Cohort 10 1-6 ND11 1000 6,000 12,000 Cohort 11 3-6 Cohort 11 1-6 ND12 1000 9,000 18,000 Cohort 12 3-6 Cohort 12 1-6 ND13 0.01 370.0 13 cohort 13 3-6 cohort 13 1-6 ND14 1000 20,000 40,000 cohort 14 3-6 cohort 14 1-6 ND15 1000 20,000 80,000 cohort 15 3-6 cohort 15 1-6 ND16 1000 20,000 160,000 cohort 16 3-6 cohort 16 1-6 ND17 1000 20,000 320,000 Cohort 17 3-6 Cohort 17 1-6

[0180] In this study, each dose level comprises an initial dose of REGN1979 followed by an escalating dose; for cohorts ND11 and above, a secondary dose has been added before reaching dose escalation (see Table 1). The initial (primary) dose, the intermediate (secondary) dose (if applicable), and the first administration of the escalating (tertiary) dose are each divided 109 optionally in at least 2 days. All patients in the ND7 and higher maximum dose cohorts received divided doses (eg, fractionated dosing) for the initial (primary) and intermediate (secondary) doses. Patients in the ND11 and higher maximum dose cohorts received divided dosing (eg, fractionated dosing) for initial (primary), intermediate (secondary), and first (tertiary) dose escalation, followed by dose escalations (weekly maximum dose). subsequently.

[0181] Study Duration: The treatment period is 9 months. Patients will be treated with up to 24 doses of REGN1979 - 4 weekly doses over a 4-week induction period, followed by an additional 8 weekly doses, and 12 or more doses given twice monthly (every 2 weeks) during a maintenance period. until progression, with the option to discontinue treatment after 9 months after the patient achieved a complete response. Patients can be monitored for efficacy and safety for up to 15 months after the end of treatment.

[0182] Study Population: Assuming enrollment through ND17 and full enrollment in all expansion cohorts, up to 370 patients are planned at approximately 15 sites in the United States and Germany. Up to 204 patients will be enrolled in the l / U I / DUO augmentation cohorts 110 the dose up to LD17 for both indications (NHL and CLL) during the dose escalation phase. Disease-specific expansion cohorts will enroll up to 100 patients comprising 90 NHL patients (20 patients with DLBCL after failure of CAR-T therapy, 40 patients with aggressive lymphoma [other than DLBCL after failure of therapy with CAR-T], 30 patients with LE grade 13a) and 10 patients with CLL. Up to 42 patients will be enrolled in the initial rituximab cohort to determine the optimal dosing regimen. Once the optimal rituximab lead-in dose regimen is determined, an additional 24 patients will be enrolled in the initial rituximab expansion and combined with the 6 patients from the previous rituximab lead-in group treated with the optimal dose and dosing regimen. Therefore, a total of 30 patients will be evaluated with the optimal starting dose and dosing regimen of rituximab.

[0183] Patients must have a documented CD20+ B-cell neoplasm, with active disease unresponsive to prior therapy, for whom no standard of care option exists, and for whom treatment with an anti-CD20 antibody may be appropriate. Patients with NHL must have been previously treated with anti-CD20 antibody therapy.

[0184] Inclusion Criteria: A patient must meet the following criteria to be eligible for inclusion A / E / ZUZ l / U I / DUO 111 in the studio: 1. Have a documented CD20+ B-cell neoplasm, with active disease unresponsive to prior therapy, for whom no standard of care option exists, and for whom treatment with an anti-CD20 antibody may be appropriate: • B-NHL Confirmed by NCI Task Force Criteria, 2007 (Cheson 2007, Appendix 2); and • CLL confirmed by the International Workshop on Chronic Lymphocytic Leukemia (IWCLL) Working Group Criteria, 2008 (Hallek 2008, Appendix 3) - Patients with small lymphocytic lymphoma (SLL) will be enrolled in the CLL group and follow the NHL evaluations. Note - A patient with a CD20-negative lymph node (NHL) biopsy performed as standard of care just prior to enrollment remains eligible for the study as long as the patient has previously documented CD20+ disease AND has been previously treated with rituximab or another antibody therapy directed against CD20 in approximately 6 months. Individual cases can be discussed with the medical monitor. 2. Patients with NHL must have received prior treatment with anti-CD20 antibody therapy. Patients with CLL need not have received prior treatment with anti-CD20 antibody therapy, as long as A / E / ZUZ l / U I / DUO 112 the patient has failed a BTK inhibitor or a PI3K inhibitor and the treating physician deems it appropriate for the patient to enter a phase 1 trial. For inclusion in the grade l-3a LF expansion cohort, patients they must have received at least 2 prior lines of systemic therapy, which includes an anti-CD20 antibody and an alkylating agent. For inclusion in the disease-specific expansion cohort enrolling patients with DLBCL after CAR-T therapy failure, the patient must have recovered from the toxicities of lymphodepletion therapy and CAR-T infusion . Prior CAR-T therapy need not be the most recent line of therapy prior to study enrollment. 3. All patients (B-cell NHL and CLL) must have at least one bidimensionally measurable lesion >1.5 cm) documented by computed tomography or magnetic resonance imaging, if computed tomography is not feasible. 4. Patients with CLL must have white blood cells (WBC) <200 x 109 / L 5. Age >18 years 6. Eastern Cooperative Oncology Group (ECOG) Performance Status <1 7. Life expectancy of at least 6 months 8. Adequate bone marrow function documented by: a. A / C / ZUZ l / U I / DUO 113 Platelet count >75 x 109 / L; b. Hb level >9 g / dL; c. ANC >1 x 109 / L Note - Patients with cell counts below the thresholds listed above may be considered for enrollment if, in the Investigator's opinion, the reason is believed to be due to bone marrow infiltration from underlying disease. In such cases, the investigator must discuss eligibility with the sponsor and receive approval for enrollment in writing. 9. Adequate organ function documented by: • Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) <2.5 X ULN • Total bilirubin <1.5 X ULN Note - Patients with Gilbert's syndrome do not need to meet this requirement as long as their total bilirubin does not change from baseline. • Cockcroft-Gault creatinine clearance >50 mL / min Note - Patients may be considered for enrollment if, in the opinion of the Investigator, the abnormal laboratory results are due to underlying disease. In such cases, the investigator must discuss eligibility with the sponsor and receive approval for enrollment in writing. Note - Patients with creatinine clearance at the A / E / ZUZ l / U I / DUO 114 Cockcroft-Gault cutoffs may be considered for enrollment if measured creatinine clearance (based on 24-hour urine or other reliable method) is >50 mL / min. 10. Willingness to undergo a mandatory tumor biopsy prior to treatment, if in the investigator's opinion, the patient has an accessible lesion that would require biopsy without significant risk to the patient. 11. Be willing and able to comply with clinical visits and study-related procedures 12. Provide signed informed consent.

[0185] Exclusion Criteria: A patient who meets any of the following criteria will be excluded from the study: 1. Primary central nervous system (CNS) lymphoma or known or suspected CNS involvement from non-primary CNS NHL 2. History of relevant or current CNS pathology, such as • Epilepsy, seizures, paresis, aphasia, apoplexy, severe brain injury, cerebellar disease, organic brain syndrome, psychosis, or • Evidence of the presence of inflammatory lesions and / or vasculitis on brain MRI 3. Standard antineoplastic chemotherapy (non-biological) A / E / ZUZ l / U I / DUO 115 within 5 times the half-life or within 28 days, whichever is shorter, prior to the first study drug administration. 4. Standard radiation therapy within 14 days of the first study drug administration. Note - Palliative radiation therapy to a symptomatic lymph node / lesion is permitted provided the irradiated lesion(s) or node(s) are not included as a target lesion for tumor evaluations 5. Allogeneic stem cell transplant 6. Treatment with rituximab, alemtuzumab, or other commercial or investigational biologic agent within 12 weeks prior to first study drug administration. Note - For patients with aggressive lymphoma requiring immediate treatment, the washout period may be reduced to 28 days. This will require a discussion with the sponsor and written approval. 7. Immunosuppressive therapy (other than biological) within 28 days of the first study drug administration. . Treatment with an investigational nonbiologic agent within 28 days of the first study drug administration. 9. History of allergic reactions attributed to compounds of similar chemical or biological composition to l / U I / DUO 116 study drug. 10. History of hypersensitivity to any compound in the tetracycline group of antibiotics. 11. Concurrent active neoplasm for which the patient is receiving treatment. 12. Known active bacterial, viral, fungal, mycobacterial or other infection or any significant episode of infection requiring hospitalization or treatment with intravenous anti-infectives within 4 weeks of first administration. 13. Evidence of a significant concurrent illness or medical condition that could interfere with the conduct of the study, or place the patient at significant risk including, but not limited to, significant cardiovascular disease (for example, class III or IV heart disease New York Heart Association, myocardial infarction within the past 6 months, unstable arrhythmias or unstable angina) and / or significant pulmonary disease (eg, obstructive pulmonary disease and history of symptomatic bronchospasm). Note - Patients with a medical history of cardiac disease should be evaluated by ECHO or multigroup acquisition (MUGA) scanning prior to the first administration of REGN1979 to ensure proper cardiac function and reserves. A / C / ZUZ l / U I / DUO suitable. 117 14. Continuous treatment with systemic corticosteroids, with the exception of the use of corticosteroids for other indications (non-tumor and non-immunosuppressive) up to a maximum of 10 mg / day of prednisone or equivalent. 15. Human immunodeficiency virus (HIV) infection or chronic infection with hepatitis B virus (HBV) or hepatitis C virus (HCV). Patients with hepatitis B (HepBsAg+) who have a controlled infection (serum hepatitis B virus DNA that is below the limit of detection AND who are receiving antiviral therapy for hepatitis B) may consult with their infection management physician. . 16. Known hypersensitivity to both allopurinol and rasburicase. 17. Pregnant or lactating women. 18. Women of childbearing potential* who are unwilling to use highly effective contraception prior to initial study drug treatment, during the study, and for at least 6 months after the last dose. Highly effective contraceptive measures include stable use of combined (containing estrogen and progestogen) hormonal contraceptives (oral, intravaginal, transdermal) or progestogen-only hormonal contraceptives (oral, injectable, implantable) associated with inhibition of ovulation initiated 2 or more menstrual cycles before screening; intrauterine device; system 118 intrauterine that releases hormones; bilateral tubal ligation; vasectomized couple; and or sexual abstinence!, i. * Postmenopausal women must be amenorrheic for at least 12 months to not be considered of childbearing potential. Pregnancy tests and contraception are not necessary for women with a documented hysterectomy or tubal ligation. t Sexual abstinence is considered a highly effective method only if it is defined as abstaining from heterosexual intercourse during the entire period of risk associated with the study treatment. The reliability of sexual abstinence should be assessed in relation to the duration of the clinical trial and the preferred and habitual lifestyle of the patient. i Periodic withdrawal (calendar, symptothermal, postovulatory methods), withdrawal (coitus interruptus), spermicides only, and the lactational amenorrhea (LAM) method are not acceptable methods of contraception. The female condom and the male condom should not be used together. 19. Administration of live vaccines within 28 days of first study drug administration 20. Member of the clinical site study team and / or her immediate family, unless prior approval is granted by the Sponsor.

[0186] Treatments: REGN1979 was supplied as a 119 liquid in single-use sterile vials. Each vial contained REGN1979 at a concentration of 2 mq / mL. Detailed instructions for preparation and administration were provided to the centers in the pharmacy manual. Diluent was supplied for preparation of study drug REGN1979.

[0187] Patients received REGN1979 weekly for a 4-week lead-in period, followed by another 8 weekly doses, and doses every 2 weeks until progression, at one dose per their assigned cohort.

[0188] In the initial rituximab and expansion only cohort, a single dose of rituximab (375 mg / m2) was administered one day prior to the first dose of REGN1979 [ie, study day (-1)]. REGN1979 was started at Week 1 Day 1, and the treatment period for REGN1979 was 9 months. Patients were treated with up to 24 doses of REGN1979: 4 weekly doses over a 4-week lead-in period, followed by an additional 8 weekly doses, and 12 or more doses given every 2 weeks during a maintenance period, until progression. In the initial rituximab cohort, REGN197 9 was administered using a 160 mg entry dose during weekly treatment and 320 mg during every 2-week maintenance treatment. Dose groups with escalating doses of REGN1979 below 160 mg weekly treatment and below 320 mg REGN1979 as treatment may also be evaluated. A / E / ZUZ l / U I / DUO 120 maintenance every 2 weeks. Subsequently, an additional 24 patients were evaluated with the optimal dose and dosing regimen; Together with the 6 patients in the initial rituximab group treated with the optimal dose and dosing regimen, a total of 30 patients were reviewed for safety and tolerability.

[0189] Evaluation criteria

[0190] Primary: The primary endpoints were safety (specifically, adverse events [AEs] and DLTs) to determine the maximum tolerated dose (MTD) and / or optimal biological dose (BOD) as the recommended dose of the phase 2 (DRF2) of REGN1979; and efficacy as measured by objective response rate (ORR) in the expansion cohort of patients with DLBCL after CART failure, expansion cohorts 1 and 2 in aggressive lymphoma (other than DLBCL after CART failure). therapy with CART), the expansion cohort in FL grade I-3a, and the CLL expansion cohort.

[0191] Secondary: Secondary endpoints were: • Pharmacokinetics: REGN1979 concentration • Immunogenicity: anti-REGN1979 antibodies • Antitumor activity: - Objective response rate (ORR) • Evaluation of the tumor response according to the Criteria NCI International Working Group (NCI-WG) Revised Response Guidelines for Malignant Lymphoma 121 • Assessment of tumor response according to the International Workshop on Chronic Lymphocytic Leukemia Guidelines for the Diagnosis and Treatment of CLL • For patients enrolled in the cohorts of NHL expansion, assessment of tumor response according to the Lugano Classification - Progression-free survival (PFS) and overall survival (OS) - Minimal residual disease (MRD) for patients with CLL

[0192] Exploratory endpoints include: • Pharmacodynamic (PD) measures including: - Subsets and phenotypes of B cells and T cells - Circulating cytokine levels -CRP - Changes in gene expression in peripheral blood

[0193] Procedures and evaluations

[0194] Initial procedures: brain MRI, electrocardiogram (ECG), human immunodeficiency virus (HIV), hepatitis C virus (HCV) and hepatitis B virus (HBV) tests, and coagulation. A / E / ZUZ l / U I / DUO Safety procedures: medical history, exam 122 physical, assessment of symptoms, assessment of functional status, clinical laboratory tests, vital signs, AE, and concomitant medication.

[0195] Efficacy Procedures: Tumor evaluations, including CT or MRI scans, 18Ffluorodeoxyglucose positron emission tomography-positron emission tomography (FDGPET), bone marrow aspirate and biopsies, lymph node biopsies, and / or tumors, and peripheral blood samples (CLL patients only). Blood samples were collected for PK and anti-drug antibodies (ADA) evaluation. Biomarker samples were collected to monitor changes in cytokine production, serum levels of proinflammatory cytokines, and changes in lymphocyte subsets and activation status. In addition, these samples allowed for somatic or tumor genetic analyzes to detect variations that impact the clinical course of the underlying disease or modulate the side effects of treatment.

[0196] Statistical plan: The study design was based on a traditional 3+3 design with 3-6 patients per ND for NHL patients, and a modified 3+3 design with an accelerated titration component with 1-6 patients. by ND for patients with CLL. The exact number of patients A / E / ZUZ l / U I / DUO 123 enrolled will depend on the number of patients (NHL and CLL) observed with protocol-defined LDN and grade 2 or higher treatment-related toxicity in which acute effects (with the exception of associated laboratory abnormalities) resolve within grade < 1 or baseline within 72 hours (LLC during accelerated titration component), and need to expand currently defined NDs, or open additional cohorts to lower NDs.

[0197] Patient enrollment is ongoing and up to 370 patients are planned. Up to 204 patients will be enrolled in the dose-escalation cohorts through LD17 for both indications (NHL and CLL) during the dose-escalation phase. Patients will be enrolled in disease-specific expansion cohorts, up to 100 patients comprising 90 NHL patients (20 patients with DLBCL after failure of CAR-T therapy, 40 patients with aggressive lymphoma [other than DLBCL after the failure of therapy with CAR-T] and 30 patients with FL grade l-3a) and 10 with CLL. Up to 42 patients will be enrolled in the initial rituximab cohort to determine the optimal dosing regimen. An additional 24 patients will be enrolled in an initial rituximab expansion and combined with the 6 patients from the previous rituximab initial cohort treated with the optimal dosage and dosing regimen. Therefore, a total of 30 will be evaluated. A / E / ZUZ l / U I / DUO 124 patients on the optimal starting regimen and dose of rituximab.

[0198] Data is summarized using only descriptive statistics. In general, data is summarized by ND, and by indication (NHL or LLC). Within the NHL indication, data will also be summarized by subgroups and doses of DLBCL after CAR-T therapy failure, aggressive lymphoma (other than DLBCL after CAR-T therapy failure) Cohorts 1 and 2 , and EL grade l-3a. Within the NHL indication, data are also summarized by indolent and aggressive NHL subgroups. Demographic and baseline characteristics are summarized descriptively by group.

[0199] Summaries and safety analyzes were performed on the Safety Analysis Set (SAF). The primary safety analysis was based on treatment-emergent AEs (TEAEs). This analysis comprised the basis on which conclusions regarding the safety profile of REGN1979 were drawn. All AEs reported in this study were coded using the currently available version of the Medical Dictionary for Regulatory Activities (MedDRA®). The coding will be done with the terms of the lowest level. Verbatim text, preferred term (TP), and primary system organ class (SOC) were listed. Endpoint analysis for efficacy l / U I / DUO 125 baseline and baseline values ​​is performed in the efficacy analysis set (FAS). Efficacy analyzes for expansion cohorts comprising DLBCL after CAR-T therapy failure, aggressive lymphoma (other than DLBCL after CAR-T failure) cohorts 1 and 2, FL grade I-3a , and LLC will be performed separately after all patients in the respective cohort have completed the 24-week visit or discontinued the study before this time.

[0200] Results: The results of patients given a maximum weekly dose of 5 to 320 mg indicate a low incidence of CRS. Serum concentrations of REGN1979 in patients receiving a maximum weekly dose of as little as 12 mg were observed to approach or exceed serum concentration levels that have been shown to be effective in Raj i tumor xenograft mouse models (not shown). show the data). Patients receiving a maximum weekly dose of as little as 40 mg maintained a serum concentration exceeding the minimum concentration level (2000 pg / L) shown to be effective in Raji tumor xenograft mouse models (data not shown). ). In addition, numerous partial and complete responses were also observed in patients treated at these levels, as shown in Tables 2-9, below. The accumulated safety of the REGN1979 and the experience l / U I / DUO 126 of PK through the NDT evaluation period of cohort 13N (27,000 mcg of REGN1979) demonstrates that the management algorithm for SLC or IRR reactions (ie, incremental dose escalation, split dosing during the first weeks of administration of REGN1979, and corticosteroid premedication) has been shown to be effective in preventing severe CRS or IRR despite incremental increases in dosage in successive dose cohorts. Split dosing provided a patient safety benefit at weeks 1 to 4 (available data), where fewer overall incidents of serious CRS / IRR were observed. In particular, the dosing strategy discussed in the present description provided a safer strategy to increase doses to levels above 80 mg, even 160 mg or more, with fewer serious events occurring at weeks 3 and 4 when higher doses high levels reached and exceeded the target serum concentrations discussed above. Figure 1 illustrates the incidence of CRS / IRR for patients receiving up to a maximum dose of 320 mg. To date, no patient has discontinued treatment due to a CRS / IRR adverse event. A / E / ZUZ l / U I / DUO Table 2. Response observed in Grade 13a Follicular Lymphoma. CD20xCD3t <5 mg (N=7) 5-12 mg (N=5) 18-40 mg (N=6) 160 mg (N=1) Overall response rate, n (%) 1 (14.3) 5(100 ) 5 (83.4) 1 (100) Complete response, n (%) 1 (14.3) 4 (80) 4 (66.7) 0 Partial response, n (%) 0 1 (20) 1 (16.7) 1 (100) Disease stable, n (%) 4 (57.1) 0 1 (16.7) 0 Progressive disease, n (%) 2 (28.6) 0 0 0 Duration of response, median (95% CI), months 5.3 N / A N / A (5.75 - not reached) 11.8 (4.37-11.83) N / A A / E / ZUZ l / U I / DUO+ No patient received an 80 mg dose of REGN1979

[0201] After data cut-off (Table 2), two additional evaluable patients showed complete responses (CR), one at 40 mg and the second at 320 mg. 128 Table 3. Response observed in Diffuse B-Cell Lymphoma. Big Λ / tZ / ZUZ l / U I / DUO CD20xCD3 <5 mg (N=15) 5-12 mg (N=11) 18-40 mg (N=11) 80 mg (N=3) 160 mg (N=3) 320 mg (N=2) Rate of overall response, n (%) 2(13.3) 2 (18.2) 6 (54.5) 3 (100) 1 (33.3) 1 (50.0) Complete response, n (%) 0 1 (9.1) 2(18.2) 3 (100 ) 1 (33.3) 1 (50.0) Partial response, n (%) 2(13.3) 1 (9.1) 4 (36.4) 0 0 0 Stable disease, n (%) 4(26.7) 4 (36.4) 3 (27.3) 0 1 (33.3) 1 (50.0) Progressive disease, n (%) 8 (53.3) 4 (36.4) 1 (9.1) 0 1 (33.3) 0 Missing / cannot be assessed, n (%) 1 (6.7) 1 (9.1) 1 (9.1) 0 0 0 Duration of response, median (95% CI), months 2.1 (1.5-2.6) N / A 4.4 (2.5-not met) N / A N / A N / A

[0202] Two of the three patients with 80 mg CR were failures on CAR T cell therapy. All complete responses noted in Table 3 were complete metabolic responses.

[0203] All CRs at the 80 mg, 160 mg, and 320 mg doses are continuous CRs on study treatment, pointing to durability of response. Table 4. Observed response in LDCLB after failure of CAR-T therapy. 129 CD20xCD3 3 mg (N=1) 27 mg (N=1) 40 mg (N=1) 80 mg (N=3) 160 mg (N=1) Overall response rate, π (%) 0 0 2 (66.7 ) Complete response, n (%) 0 0 2 (66.7) Partial response, n (%) 0 0 0 Stable disease, n (%) 0 1 (100) 0 Progressive disease, n (%) 1 (100) 0 1 (100) 1 (33.3) 1 (100) Missing / cannot be assessed, n (%) 0 0 0 l / U I / DUO Table 5. Response observed in the Lymphoma of the cells of the CD20xCD3 5-12 mg (N=1) 18-40 mg (N=1) 160 mg (N=1) Objective response (CR / PR) 1 (100%) 1 (100%) 0 Complete response 0 1 (100 %) 0 Partial response 1 (100%) 0 0 Stable disease 0 0 0 Progressive disease 0 0 0 Missing / Cannot assess 0 0 1 (100%) 130 Table 6. Response Observed in Marginal Zone Lymphoma CD20xCD3 5-12 mg (N=1) 18-40 mg (N=1) 80 mg (N=3) Objective response (CR / PR) 0 1 (100%) 2 (66.7 %) Complete response 0 0 2 ( 66.7%) Partial response 0 1 (100%) 0 Stable disease 0 0 0 Progressive disease 1 (100%) 0 1 (33.3%) Table 7. Responses in MCL Patients by Dose Level CD20xCD3 0.3 mg (N=1) 2 mg (N=1) 4 mg (N=1) 8 mg (N=1) 27 mg (N=1) 160 mg (N=1) Objective response (CR / PR) 0 1 (100%) 0 1 (100%) 1 (100%) 1 (100%) Full response 0 0 0 0 1 (100%) 1 (100%) Partial response 0 1 (100%) 0 1 (100 %) 0 0 Stable disease 0 0 0 0 0 0 Progressive disease 1 (100%) 0 1 (100%) 0 0 0 Missing / Cannot assess 0 0 0 0 0 0 131 Table 8. Responses in patients with MZL by dose level. CD20xCD3 4 mg (N=1) 5 mg (N=1) 27 mg (N=1) 80 mg (N=3) Objective response (CR / PR) 1 (100%) 0 1 (100%) 2 (66.7 %) Complete response 0 0 0 2 (66.7%) Partial response 1 (100%) 0 1 (100%) 0 Stable disease 0 0 0 0 Progressive disease 0 1 (100%) 0 1 (33.3%) Missing / Don't know can evaluate 0 0 0 0 A / E / ZUZ l / U I / DUO Table 9. Responses in patients with other NHL by level of CD20xCD3 4 mg (N=1) 12 mg (N=1) 18 mg (N=1) 27 mg (N=1) Subtype NHL LF grade unknown Waldenstrom's macroglobulinemia LF grade 3b LF grade unknown Objective response (CR / PR) 0 0 1 (100%) 0 Complete response 0 0 1 (100%) 0 Partial response 0 0 0 0 Stable disease 1 (100%) 0 0 0 Progressive disease 0 0 0 0 Missing / Cannot assess 0 1 (100 %) 0 1 (100%)

[0204] Overall, the overall activity observed in relapsed / refractory B-NHL patients heavily treated with REGN1979, including some with progression after prior CAR T cell therapy, included: 132 (i) FL Grade l-3a: 12 / 13 (92.3%) ORR; 8 / 13 CR (61.5%) at doses >5 mg; (ii) LDCBG: 4 / 6 (66.7%) ORR (all CR) at doses 80-160 mg, with two patients achieving CR after failure of CD19-directed CAR T-cell therapy; (iii) LCM: 3 / 3 responses to doses >5 mg, including a CR; (iv) LZM: response 3 / 5 to doses >5 mg, including two CRs; (v) Tolerability has been demonstrated in B-NHL patients up to doses of 320 mg weekly, with no LDN observed in B-NHL patients; (vi) Most adverse events are mild to moderate in severity; (vii) Infections were reported in 49.4% of patients (14.8% grade 3-4, with two deaths (2.5%)); (viii) No patients discontinued treatment due to CRS or neurological adverse events. Of 96 initial patients, only seven patients experienced grade 3 CRS; and (ix) Dexamethasone did not inhibit REGN1979 cytotoxicity, modestly affected the upregulation of T cell activation, and inhibited cytokine release. Example 2: Clinical evaluation of a bispecific antibody

[0205] The clinical study described below is 133 a multicenter phase 2 open-label study to evaluate the antitumor activity and safety of REGN1979, an anti-CD20 x anti-CD3 bispecific antibody, in patients with relapsed or refractory follicular lymphoma.

[0206] Objectives: The primary objective of this study is to evaluate the antitumor activity of a single agent REGN1979, as measured by the objective response rate (ORR) according to the Lugano Classification of Response in Malignant Lymphoma (Cheson, 2014 ) by an independent central review, in patients with follicular lymphoma (LE) who have relapsed or are refractory to at least 2 prior lines of systemic therapy, including an anti-CD20 antibody and an alkylating agent. The secondary objectives of this study are: (1) to assess the antitumor activity of single agent REGN1979 in patients with relapsed or refractory LE, as measured by (a) ORR according to the Lugano Classification (Cheson, 2014) as assessed by the local investigator , (b) complete response rate (CR) according to the Lugano Classification according to the independent central evaluation, and the evaluation of the local investigator, (c) progression-free survival (PFS) according to the Lugano Classification according to the independent central assessment, and local investigator assessment, (d) overall survival (OS), (e) duration of response (DOR) according to the Lugano Classification as assessed by an independent central review, and the 134 local investigator assessment, (f) disease control rate (DCR) according to the Lugano Classification as assessed by an independent central review, and local investigator assessment, (g) duration of disease control (DDC) ) according to the Lugano Classification as assessed by an independent central review, and assessment by the local investigator; (2) to assess the safety and tolerability of REGN1979; (3) to evaluate the pharmacokinetics (PK) of REGN1979; (4) to assess the immunogenicity of REGN1979; and (5) to assess the effect of the REGN1979 on quality of life, as measured by the validated instruments of the European Organization for Research and Treatment Cancer Quality of Life Questionnaire (EORTC QLQ-C30) and EuroQoL 5 Dimensions 3 Levels (EQ-5D-3L).

[0207] Study Design: The study consists of a screening period of up to 28 days, a total treatment period of up to 98 weeks including 12 weekly doses (CS) followed by dosing every 2 weeks (C2W) for up to 86 weeks , and a post-treatment follow-up period of 96 weeks.

[0208] REGN1979 is administered as a single agent intravenously (IV) at an initial (primary) dose of 1 mg, followed by an intermediate (secondary) dose of 20 mg, and subsequently a nominal (tertiary and maximum) dose. weekly) of 80 mg in a dosing regimen of 12 A / E / ZUZ l / U I / DUO 135 CS treatments, followed by 80 mg dosing of REGN1979 every 2 weeks.

[0209] The inscription follows a single arm open design.

[0210] The evaluation period begins with the signing of the informed consent form (IGF) and ends when the patient is confirmed to be eligible for the study and begins treatment, or when the patient is determined to be ineligible and is designated as a failure in the evaluation.

[0211] The treatment period begins with the initial administration of REGN1979 and consists of 12 CS infusions of REGN1979 followed by dosing every 2 weeks for 86 weeks for a total treatment period of 98 weeks of study drug dosing, unless the patient discontinues study treatment due to disease progression, initiation of subsequent lymphoma therapy, adverse events (AEs), or any other reason.

[0212] The post-treatment follow-up period will be 96 weeks after the last dose of study treatment. All patients will be followed every 12 weeks to determine survival status until death, loss to follow-up, withdrawal of patient consent for follow-up, or termination of the study by the sponsor, whichever 136 occurs first. For patients who discontinued study treatment for any reason other than disease progression, initiation of subsequent lymphoma therapy, or death, disease response will be assessed every 12 weeks during the post-study follow-up period. treatment until time of disease progression, death, initiation of subsequent lymphoma therapy, or withdrawal of patient consent for follow-up, whichever occurs first.

[0213] Study Duration: The study duration for each patient, excluding the screening period, will be approximately 194 weeks, unless the patient has disease progression or begins subsequent therapy, or until time of death, loss to follow-up, withdrawal of patient consent for follow-up, or termination of the study by the sponsor. The end of the study is defined as the last visit of the last patient.

[0214] Study Population: Up to 481 patients will be enrolled at up to 100 centers. The study population will consist of patients 18 years of age or older with previously treated grade 1 to 3a LE who have relapsed or are refractory to at least 2 prior lines of systemic therapy, including an anti-CD20 antibody and an alkylating agent. Central histopathological confirmation of the diagnosis of LE will be required A / C / ZUZ l / U I / DUO 137 before registration. Patients with grade 3b FL are not eligible. Refractory disease is defined as failure to respond to a standard regimen or progression within 6 months to the last treatment.

[0215] Inclusion Criteria: Each patient must meet the following criteria to be eligible for inclusion in the study: 1. 18 years of age or older 2. Central histopathological confirmation of the diagnosis of FL Grade 1 to 3a must be obtained prior to study enrollment. Patients with grade 3b FL are not eligible. Follicular lymphoma subtyping is based on the World Health Organization (WHO) classification (Swerdlow, 2017) . 3. Disease must be relapsed or refractory to >2 prior lines of systemic therapy, including anti-CD20 antibody and alkylating agent. In the investigator's opinion, patients should require LF therapy at the time of study enrollment. 4. Disease measurable on cross-sectional imaging (defined as at least 1 bidimensionally measurable nodal lesion >1.5 cm in greatest transverse diameter (GTD) regardless of minor axis diameter) documented by diagnostic imaging (computed tomography [CT] or magnetic resonance imaging [MRI]). A / E / ZUZ l / U I / DUO 138 5. Eastern Cooperative Oncology Group (ECOG) performance status 0 or 1. 6. Adequate bone marrow function documented by: (a) Platelet count >50 x 109 / L. A patient cannot have received a platelet transfusion in the 7 days prior to the first dose of REGN1979 to meet platelet eligibility criteria; (b) Hemoqlobin >9.0 g / dL; (c) absolute neutrophil count (ANC) >1.0 x 109 / L. A patient cannot have received granulocyte colony-stimulating factor within 2 days prior to the first dose of REGN1979 to meet the ANC eligibility criteria. 7. Adequate liver function: (a) Total bilirubin <1.5 x ULN (<3 x ULN if attributed to liver infiltration of lymphoma); (b) Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) <2.5 x ULN (<5 x ULN if attributed to liver lymphoma infiltration); (c) Alkaline phosphatase (ALP) <2.5 x ULN (<5 x ULN if attributed to lymphoma infiltration in the liver); NOTES - Regardless of the presence of lymphoma infiltration in the liver, a patient with an AST >2.5 x ULN and / or ALT >2.5 x ULN concurrent with a total bilirubin >1.5 x ULN will be excluded, and patients with Gilbert's syndrome known are not required to meet this total bilirubin requirement as long as the value does not change from A / E / ZUZ l / U I / DUO 139 to the initial value level. 8. Serum creatinine <1.5 x ULN, or creatinine clearance calculated by the Cockcroft-Gault formula >50 mL / min; NOTE: Patients with an estimated creatinine clearance <50 mL / min may be considered for enrollment if the measured creatinine clearance (based on 24-hour urine collection or other reliable method) is >50 mL / min. 9. Availability to undergo a tumor biopsy at baseline. If an investigator has determined that an initial tumor biopsy cannot be safely obtained, the sponsor may grant an exception to the biopsy requirement only after discussion and approval with the medical monitor. 10. Ability to understand the purpose and risks of the study and provide signed and dated informed consent and authorization to use protected health information (in accordance with national and local subject privacy regulations). 11. Be willing and able to comply with clinical visits and study-related procedures. 12. Provide informed consent signed by the study patient or a legally acceptable representative. 13. Able to understand and complete the questionnaires related to the study. 140

[0216] Exclusion Criteria: A patient who meets any of the following criteria will be excluded from the study: 1. Primary central nervous system (CNS) lymphoma or known non-primary CNS NHL involvement (suspected CNS lymphoma should be evaluated by lumbar puncture, as appropriate, in addition to mandatory head CT or MRI). 2. Treatment with any systemic anti-lymphoma therapy within 5 half-lives or within 28 days prior to the first study drug administration, whichever is shorter. 3. History of allogeneic stem cell transplantation. 4. Prior treatment with any chimeric antigen receptor T cell (CAR-T) therapy. 5. Continuous systemic corticosteroid therapy with more than 10 mg per day of prednisone or an equivalent anti-inflammatory within 72 hours of study drug initiation. 6. History of neurodegenerative disease or CNS movement disorder. History of uncontrolled seizure disorder, defined as any seizure within the 12 months prior to study enrollment. 7. Vaccination within 28 days prior to the first study drug administration with a vector having A / E / ZUZ l / U I / DUO 141 replication potential. 8. Other neoplasms except FL within the last 5 years, with the exception of non-melanoma skin cancer that has undergone potentially curative therapy or cervical carcinoma in situ, or any other tumor deemed to be effectively treated with definitive local control and with curative intent. 9. Evidence of significant concurrent illness or medical condition that could interfere with study performance or place the patient at significant risk, including, but not limited to, significant cardiovascular disease {for example, Class III or IV heart disease of the New York Heart Association, myocardial infarction within the previous 6 months, unstable arrhythmias, or unstable angina) and / or significant pulmonary disease (eg, obstructive pulmonary disease and history of symptomatic bronchospasm). 10. Cardiac ejection fraction <40% by echocardiogram or multi-stage acquisition (MUGA) scan. 11. Any infection requiring hospitalization or intravenous anti-infective treatment within 2 weeks of first study drug administration. 12. Uncontrolled infection with the human immunodeficiency virus (HIV), hepatitis B or hepatitis C; or other uncontrolled infection, except: (a) HIV-infected patients who have a controlled infection (load A / E / ZUZ l / U I / DUO 142 undetectable viral and CD4 count greater than 350 cells / microliter, either spontaneously or with a stable antiviral regimen); (b) Patients with hepatitis B (HepBsAg+) who have a controlled infection (serum hepatitis B virus DNA polymerase chain reaction [PCR] that is below the limit of detection AND who are receiving antiviral therapy to hepatitis B); (c) Patients who are positive for antibodies to hepatitis C virus (HCV Ab+) who have a controlled infection (HCV RNA undetectable by PCR, either spontaneously or in response to a previous successful course of hepatitis C virus) are permitted. anti-HCV therapy). 13. History of severe allergic reaction attributed to compounds with a chemical or biological composition similar to that of the drug or excipient under study. A severe allergic reaction is defined for this purpose as one that requires hospitalization and / or treatment with epinephrine. 14. Known hypersensitivity to both allopurinol and rasburicase. 15. Clinical site study team member or immediate family, unless prior approval is granted by the sponsor. 16. Women with a positive serum β-hCG pregnancy test at the screening visit. If positive, pregnancy must be ruled out by ultrasound for the patient to be eligible. A / E / ZUZ l / U I / DUO 143 17. Patients who are admitted to an institution by virtue of an order issued by the judicial or administrative authorities. 18. Pregnant or lactating women. 19. Women of childbearing potential* or men who are unwilling to use highly effective contraception prior to the initial dose / start of first treatment, during the study, and for at least 6 months after the last dose. Highly effective contraceptive measures include: (a) stable use of combined (containing estrogen and progestogen) hormonal contraception (oral, intravaginal, transdermal) or progestogen-only hormonal contraception (oral, injectable, implantable) associated with inhibition of ovulation started 2 or more menstrual cycles before screening; (b) intrauterine device (IUD); intrauterine hormone releasing system (IUS); (c) bilateral tubal ligation; (d) vasectomized couple; (e) and / or sexual abstinence, *. Postmenopausal women must be amenorrheic for at least 12 months to not be considered of childbearing age. Pregnancy tests and contraception are not necessary for women with a documented hysterectomy or tubal ligation. +Sexual abstinence is considered a highly effective l / U I / DUO method only if it is defined as abstaining from intercourse 144 heterosexuals throughout the risk period associated with the study drugs. The reliability of sexual abstinence should be assessed in relation to the duration of the clinical trial and the subject's usual and preferred lifestyle. * Periodic withdrawal (calendar, symptothermal, post-ovulation methods), withdrawal (coitus interruptus), spermicides only, and the lactational amenorrhea (LAM) method are not acceptable methods of contraception. The female condom and the male condom should not be used together.

[0217] Treatment: REGN1979 will be administered by intravenous infusion at an initial dose of 1 mg during week 1, an intermediate dose of 20 mg during week 2, and a nominal dose of 80 mg or 160 mg during subsequent administrations. For the initial dose, the intermediate dose and the first nominal dose (primary, secondary and tertiary doses, respectively), the treatments will be divided into 2 separate infusions, each for 4 hours every 2 days, preferably consecutive but not more than 3 days. of difference {for example, week 1 day 1 and week 1 day 2). Subsequent treatments (maximum weekly doses, eg 320 mg) may be given as a single infusion or as 2 separate infusions and may be given over 1 to 4 hours depending on tolerability. The study treatment comprises 12 CS administrations A / C / ZUZ l / U I / DUO 145 followed by dosing every 2 weeks for 86 weeks, for a total of 98 weeks of study drug dosing.

[0218] Endpoints: The primary study endpoint is ORR from 1st dose to 194 weeks post 1st dose, based on the Lugano Classification of Response in Malignant Lymphoma (Cheson, 2014) and according to an independent central review, in patients with FL who have relapsed or are refractory to at least 2 prior lines of systemic therapy, including an anti-CD20 antibody and an alkylating agent. Secondary endpoints are: (1) ORR according to the Lugano Classification as assessed by the local investigator from the first dose to 194 weeks after the first dose; (2) CR rate from the first dose to 194 weeks after the first dose, according to the Lugano Classification, based on independent central review, and local investigator assessment; (3) PFS from the first dose to 194 weeks after the first dose, according to the Lugano Classification, based on independent central review, and local investigator assessment; (4) OS from the first dose to 194 weeks after the first dose; (5) DOR from the first dose to 194 weeks after the first dose, according to the Lugano Classification, based on central review 146 independent and local investigator evaluation; (6) DCR from the first dose to 194 weeks after the first dose, according to the Lugano Classification, based on independent central review, and local investigator assessment; (7) DDC from the first dose to 194 weeks after the first dose, according to the Lugano Classification, based on independent central review, and local investigator assessment; (8) Incidence and severity of treatment-emergent adverse events (TEAEs) from the first dose to 194 weeks after the first dose and (9) Changes in patient-reported outcome scores from the first dose to 194 weeks. after the first dose, as measured by EORTC validated instruments QLQ-C30 and EQ-5D-3L.

[0219] Procedures and Assessments: For all patients, disease will be assessed radiologically using computed tomography (CT) or magnetic resonance imaging (MRI) and 18Ffluorodeoxyglucose (FDG) positron emission tomography (FDG) imaging. -PET). Tumor response according to the Lugano Classification criteria will be adjudicated by an independent central radiological review. A bone marrow aspirate, bone marrow biopsy, and lymph node and / or tumor biopsy will be performed, and 147 samples will be evaluated histologically, and can be used for other studies, including immunohistochemistry. Safety will be assessed by assessment of vital signs, physical examination, Eastern Cooperative Oncology Group (ECOG) performance status, electrocardiogram (ECG), incidence of AEs, and report of concomitant medication. Laboratory evaluations include complete blood count with differential, blood glucose values, serum immunoglobulins G (IgG), serum pregnancy tests (if relevant), ferritin, and C-reactive protein (CRP). Blood samples will be collected for PK and Anti-Drug Antibodies (ADA) evaluation. Peripheral blood samples will be collected to assess changes in biomarkers (eg, cytokine production, serum levels of proinflammatory cytokines, and changes in lymphocyte subsets, and activation status). In addition, these samples will allow somatic or tumor genetic analysis to detect variations that affect the clinical course of the underlying disease or modulate the side effects of treatment. Quality of life assessments will be performed using the EORTC QLQ-C30 and EQ5D-3L self-administered questionnaires.

[0220] Statistical plan: This study is designed to evaluate the efficacy and safety of REGN1979 for patients with LE who have relapsed or are refractory to at least 2 A / E / ZUZ l / U I / DUO 148 previous lines of systemic therapies. Analysis of the primary efficacy endpoint will be performed after all patients have completed the 28-week study treatment period until assessments and have had an assessment of tumor responses or have withdrawn from the study. Sample size justification: A one-stage exact binomial design is adopted for the ORR primary endpoint. Two-sided 95% confidence intervals for the observed ORRs were calculated based on the sample size 100. With 100 patients, if the observed ORR is at least 60%, 66%, 70%, and 75%, the lower limit of the 95% CI will exclude the ORR of 49%, 55%, 60% and 65% respectively; that is, the ORR is significantly different from 49%, 55%, 60%, and 65% as shown in Table 10, below. Table 10. The exact two-sided 95% confidence intervals for the observed ORR given a sample size of 100 patients. A / E / ZUZ l / U I / DUO Number of responders ORR observed 95% CI- lower 95% CI- upper 60 0.60 0.497 0.697 66 0.66 0.558 0.752 70 0.70 0.6002 0.788 75 0.75 0.653 0.831 149 With a sample size of 100 patients, if the true treatment effect of REGN1979 is 64%, 70%, 75%, or 80%, the probability is 82%, 83%, 89%, or 91% for the observed lower bound of 95% CI to exclude 49%, 55%, 60% or 65% respectively. The sample size will be further increased by 10% to account for patients who withdraw early from the study. Therefore, the total sample size will be 112 patients.

[0221] Statistical methods: Demographic and baseline characteristics will be summarized descriptively. The primary efficacy endpoint is ORR according to the Lugano Classification based on an independent central review. The ORR will be summarized along with the two-sided 95% confidence interval. Patients who are not evaluated for the best overall response will be considered as non-responders. Secondary ORR efficacy endpoints by investigator review according to the Lugano Classification, and CR and DCR rate by local investigator assessment and by independent central review according to the Lugano Classification they will be summarized along with the two-sided 95% confidence interval. The other secondary efficacy endpoints, including DOR, DDC, PFS, and OS will be summarized by the median and its 95% confidence interval using the Kaplan-Meier method according to the 150 Lugano classification. The disease control rate will be summarized together with the two-sided confidence interval. The quality of life measured by the EORTC validated instruments QLQ-C30 and EQ-5D-3L will be summarized using descriptive statistics. Observations and safety measurements, including drug exposure, AEs, laboratory data, vital signs, and ECOG performance status will be summarized and presented in tables and listings.

[0222] Interim analysis: An interim analysis will be performed after the first 50 patients who have completed the 28-week tumor assessments or have withdrawn from the study earlier. The ORR and associated 95% confidence interval will be summarized. Since the primary purpose of this interim analysis is the point estimate of the ORR and characterization of the precision of the point estimate, there are no hypothesis tests associated with this interim analysis. Therefore, the Type I error adjustment is not applicable for this planned interim analysis. For other efficacy endpoints, a two-sided 95% confidence interval will also be presented.

[0223] Further REGN1979 efficacy investigations may be included in the present study, or in additional studies, including in (a) patients with follicular lymphoma (grade l-3a) as third-line or higher (3L+) therapy, ( b) patients with follicular lymphoma (grade l-3a) who 151 are candidates for full-dose chemotherapy as second-line or higher (2L+) therapy, (c) patients with follicular lymphoma (grade l-3a) who are not candidates for full-dose chemotherapy as 2L+ therapy, (d) patients with follicular lymphoma (grade I-3a) who are previously untreated and are candidates for full-dose chemoimmunotherapy, (e) patients with follicular lymphoma (grade I-3a) who are previously untreated and are not candidates for full-dose chemoimmunotherapy full dose, (f) patients with follicular lymphoma (grade 1-3a) eligible for full-dose chemoimmunotherapy as first-line (IL) therapy versus standard of care, (g) patients with follicular lymphoma (grade 1-3a) unfit for full-dose chemoimmunotherapy immunotherapy as IL therapy versus standard of care, (h) patients with follicular lymphoma (grade l-3a) eligible for full-dose chemoimmunotherapy as 2L+ therapy versus standard of care, (i) patients with follicular lymphoma ( grade l-3a) not eligible for full-dose chemoimmunotherapy as 2L+ therapy versus standard of care and / or (j) patients with follicular lymphoma in combination with standard of care.

[0224] Further REGN1979 efficacy investigations may be included in the present study, or in additional studies, including in (a) patients with de novo or transformed diffuse large B-cell lymphoma (DLBCL) as 3L+ therapy, (b) patients with DLBCL after failure of the A / E / ZUZ l / U I / DUO 152 CAR-T therapy, (c) DLBCL patients eligible for autologous HSCT (Hematopoietic Stem Cell Transplantation) as 2L+ therapy, (d) DLBCL patients ineligible for HSCT as 2L+ therapy, (e) no treatment DLBCL patients previous history, with poor molecular prognostic factors (non-germinal center B, double or triple hit) and suitable for full-dose chemoimmunotherapy, (f) patients with DLBCL who have not previously been treated, poor molecular prognostic factors (non-germinal center B , double or triple hit) and not suitable for full-dose chemoimmunotherapy, (g) patients with DLBCL who have not received CAR-T, (h) patients with DLBCL with a maximum weekly dose of 320 mg, (i) patients with DBLK in combination with standard of care, (j) patients with DLBCL for full-dose GIM as 1L therapy versus standard of care, (k) patients with DLBCL not eligible for full-dose GIM as 1L therapy versus standard of care, (1) patients with DLBCL who are eligible for self-HSCT as 2L+ therapy versus standard of care, and / or (m) patients with DLBCL who are not eligible for self-HSCT as 2L+ therapy versus standard of care.

[0225] Further REGN1979 efficacy investigations may be included in the present study, or in further studies, including in (a) patients with MCL after BTK inhibitor failure as 2L+ therapy, (b) in patients with lymphoma of the A / E / ZUZ l / U I / DUO 153 marginal zone (MZL) as 2L+ therapy, and / or (c) in patients with lymphoblastic lymphoma, lymphoplasmacytic lymphoma, Burkitt's lymphoma, or other B-NHL subtypes as 2L+ therapy.

[0226] Additional REGN1979 efficacy investigations may be included in the present study, or in additional studies, including in (a) patients with a CD20+ B-cell neoplasm who have received a single dose of rituximab one day prior to the first dose of REGN1979. In this initial and expansion cohort of rituximab only a single dose of rituximab (375 mg / m2) will be administered one day before the first dose of REGN1979 [ie, the day of the study (-1)]. REGN1979 will start at Week 1 Day 1, and the treatment period for REGN1979 will be 9 months. Patients will be treated with up to 24 doses of REGN1979: 4 weekly doses over a 4-week lead-in period, followed by an additional 8 weekly doses, and 12 doses given every 2 weeks over a 6-month maintenance period. In the first part of this initial rituximab cohort, REGN1979 will be administered using an 80 mg dose escalation. Once an optimal dose regimen is identified, an additional dose group of 6 patients will be evaluated with a 320 mg dose escalation of REGN1979 at the optimal dose regimen. Dose groups with REGN1979 dose escalations between 80 mg and 320 mg can also be evaluated. Subsequently, 24 patients will be evaluated A / E / ZUZ l / U I / DUO An additional 154 patients with this optimal dose regimen and the optimal dose, and together with the 6 patients in the initial rituximab group treated with the optimal dose, a total of 30 patients will be reviewed for safety and tolerability.

[0227] In any of the standard of care combination studies, the combination may include REGN1979 plus CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone), ICE (ifosfamide, carboplatin, and etoposide), GemOx (gemcitabine and oxaliplatin), lenalidomide , or lenalidomid 10 plus rituximab.

[0228] The scope of the present invention is not limited to the embodiments described herein. In fact, various modifications of the invention, in addition to those described herein, will be apparent to persons skilled in the art from the foregoing description. Such modifications are intended to fall within the scope of the appended claims.

Claims

1. A method of administering a therapeutic protein to a subject on a dosing regimen to mitigate the adverse effects of cytokine release syndrome or an infusion-related reaction, comprising: administering fractions of a primary dose (PD) of the therapeutic protein in week 1 of the dosing regimen, wherein the primary dose comprises no more than 10 mg of the therapeutic protein, a first dose fraction (F1D1) comprises 40% to 60% of the total primary dose and is administered to the subject on day 1 of week 1, and a second dose fraction (F2D1) comprises the remaining 40% to 60% of the total primary dose and is administered to the subject 12 to 96 hours after the administration of F1D1;administering fractions of a secondary dose (D2) of the therapeutic protein in week 2 of the dosing regimen, wherein the secondary dose is no more than half of a maximum weekly dose of the therapeutic protein, a first dose fraction (F1D2) comprises 40 to 60% of the total secondary dose, a second dose fraction (F2D2) comprises the remaining 40% to 60% of the total secondary dose, and F2D2 is administered to the subject 12 to 96 hours after the administration of F1D2 during week 2 of the dosing regimen; and administering the maximum weekly dose of the therapeutic protein to the subject as a single dose in a later week of the dosing regimen.

2. The method of claim 1, wherein F2D1 is administered to the subject 24 to 96 hours after the administration of F1D1.

3. The method of claim 1, wherein F2D1 is administered to the subject 18 to 72 hours after the administration of F1D1.

4. The method of any of claim 13, wherein F2D2 is administered to the subject 24 to 96 hours after the administration of F1D2.

5. The method of any of claim 13, wherein F2D2 is administered to the subject 18 to 72 hours after the administration of F1D2.

6. The method of any of claims 1-5, wherein the later week is week 3 of the dosing regimen.

7. The method of any of claims 1-5, wherein the later week is week 4 of the dosing regimen.

8. The method of any of claims 1-5, wherein the later week is week 14 of the dosing regimen.

9. The method of any of claims 1- A / C / ZUZ l / UI / DUO 157 5, wherein the later week is any of weeks 4 to 36 of the dosing regimen.

10. The method of any of claim 15, further comprising: administering fractions of a tertiary dose (D3) of the therapeutic protein in week 3 of the dosing regimen, wherein the tertiary dose is not less than half of the maximum weekly dose of the therapeutic protein and not more than the maximum weekly dose of the therapeutic protein, a first dose fraction (F1D3) comprises from 40% to 60% of the total tertiary dose, a second dose fraction (F2D3) comprises the remaining 40% to 60% of the total tertiary dose, and F2D3 is administered to the subject 12 to 96 hours after the administration of F1D3 during week 3 of the dosing regimen; and administering the maximum weekly dose of the therapeutic protein to the subject as a single dose in a subsequent week of the dosing regimen.

11. The method of claim 10, wherein F2D3 is administered to the subject 24 to 96 hours after the administration of F1D3.

12. The method of claim 10, wherein F2D3 is administered to the subject 18 to 72 hours after the administration of F1D3.

13. The method of any of claims 10 l / UI / DUO 158 12, wherein the later week is week 4 of the dosing regimen.

14. The method of any of claims 1012, wherein the later week is week 14 of the dosing regimen.

15. The method of any one of claim 1012, wherein the later week is any one of weeks 4 to 36 of the dosing regimen.

16. The method of any one of claim 1012, wherein the tertiary dose is administered as a single dose in weeks 4 to 12 of the dosing regimen.

17. A method of administering a therapeutic protein to a subject on a dosing regimen to mitigate the adverse effects of cytokine release syndrome or an infusion-related reaction, comprising: administering fractions of a primary dose (PD) of the therapeutic protein in week 1 of the dosing regimen, wherein the primary dose comprises no more than 10 mg of the therapeutic protein, a first dose fraction (F1D1) comprises 40% to 60% of the total primary dose and is administered to the subject on day 1 of week 1, and a second dose fraction (F2D1) comprises the remaining 40% to 60% of the total primary dose and is administered to the subject 12 to 96 hours after the administration of F1D1;A / C / ZUZ l / UI / DUO 159 administer fractions of a secondary dose (D2) of the therapeutic protein in week 2 of the dosing regimen, wherein the secondary dose is equal to a maximum weekly dose of the therapeutic protein, a first dose fraction (F1D2) comprises 40% to 60% of the total secondary dose, a second dose fraction (F2D2) comprises the remaining 40% to 60% of the total secondary dose, and F2D2 is administered to the subject 12 to 96 hours after the administration of F1D2 during week 2 of the dosing regimen; and administer the maximum weekly dose of the therapeutic protein to the subject as a single dose in a later week of the dosing regimen.

18. The method of any of claim 117, wherein the maximum weekly dose of the therapeutic protein is administered to the subject as a single dose for 1 to 8 weeks, 1 to 12 weeks, or 1 to 16 weeks during a weekly phase of the dosing regimen.

19. The method of any of claim 118, further comprising administering the maximum weekly dose of the therapeutic protein to the subject as a single dose once every two weeks during a maintenance phase of the dosing regimen, following the completion of a weekly phase of the dosing regimen.

20. The method of any of claims 1 A / E / ZUZ l / UI / DUO 160 18, further comprising administering the maximum weekly dose of the therapeutic protein to the subject as a single dose once every three weeks during a maintenance phase of the dosing regimen, which follows the completion of a weekly phase of the dosing regimen.

21. The method of any of claim 118, further comprising administering the maximum weekly dose of the therapeutic protein to the subject as a single dose once every four weeks during a maintenance phase of the dosing regimen, following the completion of a weekly phase of the dosing regimen.

22. The method of any of claims 1921, wherein the maintenance phase is at least 24 weeks.

23. The method of claim 22, wherein the maintenance phase is 24 weeks.

24. The method of any of claims 123, wherein the primary dose (DI) is 1 mg.

25. The method of any of claim 124, wherein the secondary dose (D2) is 20 mg.

26. The method of any of claims 1016 or 18-25, wherein the tertiary dose is 40 mg.

27. The method of any of claims 1016 or 18-25, wherein the tertiary dose is 80 mg.

28. The method of any of claims 10-16 or 18-25, wherein the tertiary dose is 160 mg. 161 29. The method of any of claims 1016 or 18-25, wherein the tertiary dose is 320 mg.

30. The method of any of claim 129, wherein F1D1 comprises 50% of the total primary dose and F2D1 comprises 50% of the total primary dose.

31. The method of any of claim 130, wherein F1D2 comprises 50% of the total secondary dose and F2D2 comprises 50% of the total secondary dose.

32. The method of any of claims 1016 or 18-31, wherein F1D3 comprises 50% of the total tertiary dose, and F2D3 comprises 50% of the total tertiary dose.

33. The method of any of claim 132, wherein the maximum weekly dose of the therapeutic protein is from 5 mg to 320 mg.

34. The method of claim 33, wherein the maximum weekly dose of the therapeutic protein is 6-320 mg, 10-320 mg, 5-40 mg, 5-80 mg, 5-160 mg, 12-40 mg, 18-80 mg, 40-80 mg, 80-160 mg, 160-320 mg, 5 mg, 6 mg, 7 mg, 8 mg, 12 mg, 18 mg, 27 mg, 40 mg, 80 mg, 160 mg, 320 mg, 480 mg or 640 mg.

35. The method of claim 34, wherein the maximum weekly dose is 80 mg.

36. The method of claim 34, wherein the maximum weekly dose is 160 mg.

37. The method of claim 34, wherein the maximum weekly dose is 320 mg.

38. The method of any of claim 137, wherein each dose or fraction of a dose is administered to the subject over a period of 1 to 6 hours.

39. The method of any of claim 138, wherein the subject has been diagnosed with cancer, for example, blood cancer.

40. The method of claim 39, wherein the cancer is a B-cell neoplasm.

41. The method of claim 40, wherein the B-cell neoplasm is a CD20+ B-cell neoplasm.

42. The method of claim 40 or 41, wherein the cancer is a non-Hodgkin lymphoma, Hodgkin lymphoma, chronic lymphocytic leukemia, acute lymphoblastic leukemia, small lymphocytic lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, marginal zone lymphoma, Burkitt lymphoma, primary mediastinal B-cell lymphoma, lymphoblastic lymphoma, or Waldenstrom macroglobulinemia.

43. The method of any of claim 138, wherein the subject has been diagnosed with follicular lymphoma (FL).

44. The method of claim 43, wherein the LE is grade l-3a.

45. The method of any of claims 1 A / E / ZUZ l / UI / DUO 163 38, wherein the subject has been diagnosed with diffuse large B cell lymphoma (DLBCL).

46. ​​The method of claim 45, wherein the subject has failed prior CAR-T therapy.

47. The method of any of claim 138, wherein the subject has been diagnosed with mantle cell lymphoma (MCL).

48. The method of claim 47, wherein the subject has failed prior therapy with Bruton's tyrosine kinase (BTK) inhibitors.

49. The method of any of claim 138, wherein the subject has been diagnosed with marginal zone lymphoma (MZL).

50. The method of claim 39, wherein the cancer is selected from pancreatic carcinoma, head and neck cancer, prostate cancer, malignant gliomas, osteosarcoma, colorectal cancer, gastric cancer, malignant mesothelioma, multiple myeloma, ovarian cancer, small cell lung cancer, non-small cell lung cancer, synovial sarcoma, thyroid cancer, breast cancer, melanomaglioma, breast cancer, squamous cell carcinoma, esophageal cancer, clear cell renal cell carcinoma, chromophobe renal cell carcinoma, oncocytoma, transitional cell carcinoma, urothelial carcinoma, bladder adenocarcinoma, or small cell carcinoma of the bladder.

51. The method of any of claims 150, wherein the subject is an adult human.

52. The method of any of claim 151, wherein the therapeutic protein is an antibody or an antigen-binding fragment thereof.

53. The method of claim 52, wherein the antibody is a fully human antibody.

54. The method of claim 52 or 53, wherein the antibody is a bispecific antibody or an antigen-binding fragment thereof.

55. The method of claim 54, wherein a first antigen-binding arm of the bispecific antibody or antigen-binding fragment binds to a T cell antigen.

56. The method of claim 55, wherein the T cell antigen is CD3.

57. The method of claim 55, wherein the T cell antigen is CD28.

58. The method of any of claims 5457, wherein the second antigen-binding arm of the bispecific antibody or antigen-binding fragment binds to a tumor cell antigen.

59. The testing method 58, where the tumor cell antigen is selected from the group that 165 consists of the proteins AFP, ALK, BAGE, BCMA, BIRC5 (survivine), BIRC7, β-catenin, brc-abl, BRCA1, BORIS, CA9, carbonic anhydrase IX, caspasa-8, CALR, CCR5, CD19, CD20 (MS4A1), CD22, CD40, CD70, CDK4, CEA, cyclin-Bl, CYP1B1, EGFR, EGFRvIII, ErbB2 / Her2, ErbB3, ErbB4, ETV6-AML, EpCAM, EphA2, Fra-1, FOLR1, GAGE ​​proteins (por example, GAGE-1, -2), GD2, GD3, GloboH, glypican-3, GM3, gplOO, Her2, HLA / B-raf, HLA / kras, HLA / MAGE-A3, hTERT, LMP2, MAGE proteins (for example, MAGE-1, -2, -3, -4, -6 and -12), MART-1, mesothelin, ML-IAP, Mucl, Muc2, Muc3, Muc4, Muc5, Mucl6 (CA-125), MUM1, NA17, NYBR1, NY-BR62, NY-BR85, NY-ESO1, pl5, p53, PAP, PAX3, PAX5, PCTA-1, PLACI, PRLR, PRAME, PSMA (FOLH1), RAGE proteins, Ras, RGS5, Rho, SART-1, SART-3, STEAP1, STEAP2, TAG-72, TGF-β, TMPRSS2, Thompson-nouvelle antigen (Tn), TRP-1, TRP-2, tyrosinase and uroplaquine-3.

60. The method of claim 59, wherein the tumor cell antigen is CD20.

61. The method of claim 60, wherein the bispecific antibody is an anti-CD20 x anti-CD3 antibody.

62. The method of claim 61, wherein the anti-CD20 x anti-CD3 antibody is REGN1979, or an antibody comprising the HCVR / LCVR of REGN1979, or an antibody comprising the CDR of REGN1979.

63. The method of claim 59, wherein the tumor cell antigen is BCMA. A / E / ZUZ l / UI / DUO 166 64. The method of claim 63, wherein the bispecific antibody is an anti-BCMA x anti-CD3 antibody.

65. The method of claim 59, wherein the tumor cell antigen is PSMA.

66. The method of claim 65, wherein the bispecific antibody is an anti-PSMA x anti-CD3 antibody.

67. The method of claim 59, wherein the tumor cell antigen is MUC16.

68. The method of claim 67, wherein the bispecific antibody is an anti-MUC16 x anti-CD3 antibody.

69. The method of claim 59, wherein the tumor cell antigen is STEAP2.

70. The method of claim 69, wherein the bispecific antibody is an anti-STEAP2 x anti-CD3 antibody.

71. The method of any of claim 170, wherein the therapeutic protein is maintained at a serum concentration of approximately 2000 mcg / L or higher after administration of the maximum weekly dose for the duration of the dosing regimen.

72. The method of claim 71, wherein the therapeutic protein is maintained at a serum concentration of approximately 2600 mcg / L or higher after administration of the maximum weekly dose 167 for the duration of the dosing regimen.

73. The method of claim 72, wherein the therapeutic protein is maintained at an average serum concentration of at least approximately 3700 mcg / L after administration of the maximum weekly dose for the duration of the dosing regimen.

74. The method of any of claim 173, wherein the therapeutic protein is administered to the subject in combination with a second agent selected from a spheroid, an antihistamine, acetaminophen, a non-steroidal anti-inflammatory drug (NSAID), an IL-6 antagonist, or an IL-6R antagonist.

75. The method of claim 74, wherein the spheroid is dexamethasone or the NSAID is indomethacin.

76. The method of claim 75, wherein dexamethasone is administered to the subject approximately one to three hours before the first fraction of the dose (F1D1).

77. The method of claim 75, wherein dexamethasone is administered to the subject approximately one to three hours before F1D1, F2D1, F1D2, and F2D2.

78. The method of claim 75, wherein dexamethasone is administered to the subject approximately one to three hours before F1D1, F2D1, F1D2, F2D2, F1D3, and F2D3. A / E / ZUZ l / UI / DUO 79. The method of claim 74, wherein the IL-6 antagonist is an anti-IL-6 antibody or the IL-6R antagonist is an anti-IL-6R antibody.

80. The method of claim 79, wherein the anti-IL-6R antibody is sarilumab.

81. The method of any of claim 7480, wherein the administration of the second agent is eliminated after a first administration of the maximum weekly dose for the duration of the dosing regimen.

82. The method of any of claim 181, wherein the therapeutic protein is administered to the subject in combination with a second therapeutic agent.

83. The method of claim 82, wherein the second therapeutic agent comprises at least one of rituximab, obinutuzumab, cyclophosphamide, doxorubicin, vincristine, prednisone, prednisolone, bendamustine, lenalidomide, chlorambucil, ibritumomab tiuxetan, idelalisib, copanlisib, duvelisib, etoposide, methylprednisolone, cytarabine, cisplatin, mesna, ifosfamide, mitoxantrone, and procarbazine.

84. The method of claim 82, wherein the second therapeutic agent comprises a combination of cyclophosphamide, doxorubicin, vincristine, and prednisone.

85. The method of claim 82, wherein the second therapeutic agent comprises a combination of ifosfamide, cisplatin, and etoposide.

86. The method of claim 82, wherein the second therapeutic agent comprises a combination of gemcitabine and oxaliplatin.

87. The method of claim 82, wherein the second therapeutic agent comprises a combination of lenalidomide and rituximab.

88. The method of claim 82, wherein the second therapeutic agent is lenalidomide.

89. A method of treating B-cell cancer in a subject, comprising: selecting a subject diagnosed with B-cell cancer; and administering a therapeutic protein to the subject according to the method of any of claim 188.

90. The method of claim 89, wherein the subject has been previously treated with cancer therapy.

91. The method of claim 89 or 90, wherein the subject is refractory to prior treatment or has relapsed after prior treatment.

92. The method of claim 89, wherein the subject has been previously treated with anti-CD20 antibody therapy.

93. The method of any of claims 89- A / E / ZUZ l / UI / DUO 92, wherein the subject has been previously treated with CAR-T therapy 170.

94. The method of any of claims 8993, wherein the B-cell cancer is selected from the group consisting of follicular lymphoma, diffuse large B-cell lymphoma, mantle cell lymphoma, and marginal zone lymphoma.

95. The method of any of claim 194, wherein the incidence of SLC and grade 3 IRR is less than 10%. 10 96. The method of claim 95, wherein the incidence of SLC and grade 3 IRR is less than 7.5% or less than 7%.

97. The method of claim 95 or 96, wherein the maximum weekly dose is 80 mg or more. 15 98. The method of any of claims 1-97, wherein any dose administered as a single dose is administered in no more than 1 hour.