RU2799529C2 - Dosing strategy reducing cytokine release syndrome for cd3/c20 bispecific antibodies - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: group of inventions is described, including a method for administering a therapeutic protein to a subject according to a dosing schedule to reduce the incidence or severity of cytokine release syndrome or an infusion reaction in the treatment of CD20-expressing B-cell lymphoma (versions) and a method of the treatment of CD20-expressing B-cell lymphoma in a subject.

EFFECT: invention expands the arsenal of methods of the treatment of CD20-expressing B-cell lymphoma.

84 cl, 1 dwg, 10 tbl, 2 ex

Description

LINK TO SEQUENCE LISTING

[0001] Included in this application by reference is a sequence listing provided in machine-readable form as file 10496WO01-Sequence created August 30, 2019 and having a size of 162944 bytes.

FIELD OF TECHNOLOGY

[0002] The present invention is in the medical field and relates to dosing strategies and regimens for administering therapeutic antibodies (e.g., bispecific antibodies targeting T cells) that reduce the incidence and severity of cytokine release syndrome or infusion reaction in patients undergoing immunotherapy.

BACKGROUND OF THE INVENTION

[0003] Cytokine release syndrome (CRS) is a systemic inflammatory response that can be caused by a variety of factors, including some drugs. Cancer immunotherapies that activate T cells are associated with a particularly high risk of CRS, which is usually associated with target effects induced by binding of a bispecific antibody or chimeric antigen receptor (CAR) T cell to their antigen and subsequent activation of third-party immune cells and non-immune cells such as endothelial cells. Activation of foreign cells leads to massive release of a number of cytokines. IL-6, IL-10, and interferon (IFN)-ɣ are among the major cytokines that are persistently elevated in the serum of patients with CRS. When using T-cell activating therapies directed against tumor cells, CRS is triggered by a massive release of IFN-ɣ by activated T cells or by the tumor cells themselves. Secreted IFN-ɣ induces the activation of other immune cells, primarily macrophages, which in turn produce excess 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 extravasation and activation of the complement and coagulation cascade causing disseminated intravascular coagulation syndrome. In addition, IL-6 probably contributes to the development of cardiomyopathy by causing myocardial dysfunction. Shimabukaro-Vornhagen et al. , Journal for Immunotherapy of Cancer , 6:56, pp. 1-14, 2018. In some cases, symptoms associated with CRS are referred to as an infusion reaction (IR) if they occur less than six hours after the start of the infusion, and CRS, if they appear six hours or more after the start of the infusion.

[0004] Controlling the toxic properties of cancer immunotherapy is a complex clinical challenge. Reducing CRS or IR is a key feature of certain therapies, such as CAR T cells and bispecific antibodies targeting T cells. Mild CRS is usually treated symptomatically with antihistamines, antipyretics, and fluids. Severe CRS can be a life-threatening adverse event that requires prompt and intensive treatment. Reducing the tumor burden, limiting the dose of therapy, and premedication with steroids reduces the incidence of severe CRS, as does the use of anticytokine drugs. Tocilizumab, an anti-IL-6 antibody, has become the standard of care for initial treatment of severe CRS in some settings. However, the use of dose and therapy restrictions to minimize cytokine activity can be detrimental to the effectiveness of immunotherapy. Thus, there remains a need for alternative strategies to reduce the potentially life-threatening manifestations of CRS without negatively impacting the therapeutic benefits of immunotherapy.

BRIEF DESCRIPTION OF THE INVENTION

[0005] In one aspect, the present invention relates to a method of administering to a subject a therapeutic protein according to a dosing schedule to reduce the undesirable effects of a cytokine release syndrome or infusion reaction, comprising: (i) administering portions of the primary dose (D1) of said therapeutic protein at week 1 of the dosing regimen, where the specified primary dose is not more than 10 mg of the specified therapeutic protein, the first part of the dose (F1D1) is from 40% to 60% of the total primary dose and is administered to the subject on day 1 of week 1, and the second part of the dose (F2D1) is the remaining 40 % to 60% of the total primary dose and is administered to the subject 12-96 hours after administration of F1D1; (ii) administering portions of a secondary dose (D2) of said therapeutic protein at week 2 of the dosing regimen, where said secondary dose is no more than half of the maximum weekly dose of said therapeutic protein, the first portion of the dose (F1D2) being between 40% and 60% of the total secondary dose , the second portion of the dose (F2D2) represents the remaining 40% to 60% of the total secondary dose, and F2D2 is administered to the subject 12-96 hours after F1D2 administration in week 2 of the dosing regimen; and (iii) administering to the subject the specified maximum weekly dose of the specified therapeutic protein in the form of a single dose on one of the subsequent weeks of the dosing regimen.

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

[0007] In some embodiments, the method further comprises: (i) administering fractions of a tertiary dose (D3) of said therapeutic protein at week 1 of the dosing regimen, wherein said tertiary dose is at least half of said maximum weekly dose of said therapeutic protein and not more than said maximum weekly dose of said therapeutic protein, the first dose portion (F1D3) is 40% to 60% of the total tertiary dose, the second portion of the dose (F2D3) is the remaining 40% to 60% of the total tertiary dose, and F2D3 is administered to the subject 12 to 96 hours later after administration of F1D3 at week 1 of the dosing regimen; and (ii) administering to the subject the specified maximum weekly dose of the specified therapeutic protein in the form of a single dose on one of the subsequent weeks of the dosing regimen.

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

[0009] In some instances, said subsequent week is week 4 of the dosing regimen. In some instances, the subsequent week indicated is week 14 of the dosing regimen. In some instances, said subsequent week is any of weeks 4-36 of the dosing regimen. In various embodiments, the tertiary dose is administered as a single dose at weeks 4-12 of the dosing regimen.

[0010] In one aspect, the present invention relates to a method of administering to a subject a therapeutic protein according to a dosing schedule to reduce the undesirable effects of a cytokine release syndrome or infusion reaction, comprising: (i) administering portions of the primary dose (D1) of said therapeutic protein at week 1 of the dosing regimen, where the specified primary dose is not more than 10 mg of the specified therapeutic protein, the first part of the dose (F1D1) is from 40% to 60% of the total primary dose and is administered to the subject on day 1 of week 1, and the second part of the dose (F2D1) is the remaining 40 % to 60% of the total primary dose and is administered to the subject 12-96 hours after administration of F1D1; (ii) administering portions of a secondary dose (D2) of said therapeutic protein at week 2 of the dosing regimen, where said secondary dose is equal to the maximum weekly dose of said therapeutic protein, the first portion of the dose (F1D2) being 50% of the total secondary dose, the second portion of the dose (F2D2) is 50% of the total secondary dose, and F2D2 is administered to the subject 12-96 hours after F1D2 administration at week 2 of the dosing schedule; and (iii) administering to the subject the specified maximum weekly dose of the specified therapeutic protein in the form of a single dose on one of the subsequent weeks of the dosing regimen.

[0011] In one aspect, the present invention relates to a method of administering to a subject a therapeutic protein according to a dosing regimen to reduce the undesirable effects of a cytokine release syndrome or infusion reaction, comprising: (i) administering portions of the primary dose (D1) of said therapeutic protein at week 1 of the dosing regimen, where the specified primary dose is not more than 10 mg of the specified therapeutic protein, where D1 is administered to the subject in several parts of the dose (for example, F1D1, F2D1, F3D1, F4D1, F5D1) on consecutive days during week 1; (ii) administering portions of a secondary dose (D2) of said therapeutic protein at week 2 of the dosing regimen, wherein said secondary dose is equal to or less than the maximum weekly dose of said therapeutic protein and is administered to the subject in multiple portions (e.g., F1D2, F2D2, F3D2, F4D2, F5D2 ) on consecutive days in week 2; and (iii) administering to the subject the specified maximum weekly dose (MD) of the specified therapeutic protein in the form of several parts of the MD or as a single dose in one of the subsequent weeks of the dosing regimen.

[0012] In some embodiments, the second portion of the dose (F2) is administered to the subject 12-96 (e.g., 24-72) hours after the first portion of the dose (F1), optionally, the third portion of the dose (F3) is administered to the subject at least 24 hours after the second part of the dose (F2), optionally, the fourth part of the dose (F4) is administered to the subject at least 24 hours after the third part of the dose (F3), and optionally, the fifth part of the dose (F5) is administered to the subject at least than 24 hours after the fourth dose (F4) at week 1, 2, or 3 of the dosing regimen.

[0013] In some embodiments, the methods of the present invention further comprise administering one or more "maintenance" doses during the maintenance phase of the dosing regimen following completion of the weekly phase of the regimen. In some cases, each maintenance dose is administered 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 to 8 weeks, 1 to 12 weeks, or 1 to 16 weeks during the 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 every other week during the maintenance phase of the dosing regimen following the completion of the 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 the maintenance phase of the dosing regimen following the completion of the 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 every four weeks during the maintenance phase of the dosing regimen following the completion of the weekly phase of the dosing regimen. In some embodiments, the maintenance phase is up to 86 weeks. In some embodiments, the maintenance phase is up to 87 weeks. In some embodiments, the maintenance phase is up to 88 weeks. In some embodiments, 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 embodiments, the maintenance phase is at least 24 weeks. In some embodiments, the maintenance phase is 24 weeks.

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

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

[0017] In some cases, the maximum weekly dose (MD) of a therapeutic protein is between 5 mg and 320 mg. In various embodiments, 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, 80-160mg, 160-320mg, 5mg, 6mg, 7mg, 8mg, 12mg, 18mg, 27mg, 40mg, 80mg, 160mg or 320mg. In some embodiments, the maximum weekly dose is 80 mg. In some embodiments, the maximum weekly dose is 160 mg. In some embodiments, the maximum weekly dose is 320 mg.

In some cases, the maintenance dose of the therapeutic protein is between 5 mg and 320 mg. In various embodiments, the maintenance 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 -160mg, 160-320mg, 5mg, 6mg, 7mg, 8mg, 12mg, 18mg, 27mg, 40mg, 80mg, 160mg or 320mg. In some embodiments, the maintenance dose is 80 mg. In some embodiments, the maintenance dose is 160 mg. In some embodiments, the maintenance dose is 320 mg.

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

[0019] In some embodiments, the subject is diagnosed with cancer. In some cases, the cancer is a B-cell malignancy. In some cases, the B-cell malignancy is a CD20+ B-cell malignancy. In some cases, the cancer is 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, Waldenström macroglobulinemia, 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, melanoma, glioma, 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 Bladder. In some embodiments, the subject is diagnosed with follicular lymphoma (FL). In some cases, PL has a degree of 1-3a. In some embodiments, the subject is diagnosed with diffuse large B-cell lymphoma (DLBCL). In one embodiment, the subject is diagnosed with relapsed/refractory DLBCL. In some cases, a subject diagnosed with DLBCL has failed prior CAR-T therapy. In some embodiments, the subject is diagnosed with mantle cell lymphoma (MCL). In some cases, a subject diagnosed with MCL has failed prior therapy with a Bruton's tyrosine kinase (BTK) inhibitor. In some embodiments, the subject is diagnosed with marginal zone lymphoma (MLZ).

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

[0021] In various embodiments, the therapeutic protein is an antibody or antigen-binding fragment thereof. In some cases, the antibody is a fully human antibody. In some instances, 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 a tumor cell antigen. In some cases, the tumor cell antigen is selected from the group consisting of AFP, ALK, BAGE proteins, BCMA, BIRC5 (survivin), BIRC7, β-catenin, brc-abl, BRCA1, BORIS, CA9, carbonic anhydrase IX, caspase-8, CALR , CCR5, CD19, CD20 (MS4A1), CD22, CD40, CD70, CDK4, CEA, cyclin B1, CYP1B1, EGFR, EGFRvIII, ErbB2/Her2, ErbB3, ErbB4, ETV6-AML, EpCAM, EphA2, Fra-1, FOLR1 , GAGE proteins (e.g. GAGE-1, -2), GD2, GD3, GloboH, glypican-3, GM3, gp100, Her2, HLA/B-raf, HLA/k-ras, HLA/MAGE-A3, hTERT, LMP2, MAGE proteins (e.g. MAGE-1, -2, -3, -4, -6 and -12), MART-1, mesothelin, ML-IAP, Muc1, Muc2, Muc3, Muc4, Muc5, Muc16 (CA -125), MUM1, NA17, NY-BR1, NY-BR62, NY-BR85, NY-ESO1, p15, p53, PAP, PAX3, PAX5, PCTA-1, PLAC1, PRLR, PRAME, PSMA (FOLH1), proteins RAGE, Ras, RGS5, Rho, SART-1, SART-3, STEAP1, STEAP2, TAG-72, TGF-β, TMPRSS2, Thompson-nouvelle antigen (Tn), TRP-1, TRP-2, tyrosinase and uroplakin- 3.

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

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

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

[0025] In some embodiments, the tumor cell antigen is MUC16. In some instances, 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 serum concentration of the therapeutic protein is maintained at or above about 2,000 micrograms/liter (µg/L) following the maximum weekly dose throughout the dosing regimen. In some instances, the serum concentration of the therapeutic protein is maintained at or above about 2600 μg/L after the maximum weekly dose has been administered throughout the dosing regimen. In some embodiments, the serum concentration of the therapeutic protein is maintained at or above about 3700 µg/L following the maximum weekly dose throughout 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 (NSAID), an IL-6 antagonist, or an IL-6R antagonist. In some cases, the steroid is dexamethasone. In some cases, the NSAID is indomethacin. In some instances, 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 various embodiments, the implementation of the introduction of the second agent is stopped after the first administration of the maximum weekly dose throughout 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 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, idelicib, copanlisib, duvelisib, etoposide, methylprednisolone, cytarab ina, cisplatin, mesna, ifosfamide, mitoxantrone and procarbazine. In some cases, the second therapeutic agent includes a combination of cyclophosphamide, doxorubicin, vincristine, and prednisone. In some cases, the second therapeutic agent includes a combination of ifosfamide, cisplatin and etoposide. In some cases, the second therapeutic agent includes a combination of gemcitabine and oxaliplatin. In some cases, the second therapeutic agent includes 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 B cell cancer in a subject, comprising: (a) selecting a subject diagnosed with B cell cancer; and (b) administering to said subject a therapeutic protein in accordance with any of the methods discussed above or herein using a dosing regimen to reduce the undesirable effects of a cytokine release syndrome or infusion reaction. In some embodiments, the subject has previously received treatment with an anti-CD20 antibody. In some embodiments, the subject has previously received therapy using CAR-T. 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 embodiments discussed above or herein, the incidence of grade 3 CRS and IR is less than 10%. In some cases, the incidence of CRS grade 3 and IR is less than 7.5% or less than 7%. In some embodiments, the maximum weekly dose is 80 mg or more when the incidence of CRS and IR is less than 10%, less than 9%, less than 8%, less than 7.5%, or less than 7%. In any of the embodiments, the implementation of any dose administered in the form of a single dose, you can enter 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 within the scope of the present invention. Any particular meaning discussed above or herein may be combined with another related meaning discussed above or herein denoting a range in which said values represent the upper and lower limits of the range, and such ranges are within the scope of the present invention. A therapeutic protein for use in any of the methods discussed herein, or the use of a therapeutic protein for the manufacture of a medicament for use in any of the methods discussed herein, is also within the scope of this invention.

[0033] Other embodiments will become apparent from the review of the following detailed description of the invention.

BRIEF DESCRIPTION OF GRAPHICS

[0034] FIG. 1 shows the incidence of CRS/IR during the first five weeks of REGN1979 therapy at various dose levels.

DETAILED DESCRIPTION OF THE INVENTION

[0035] Before reading the description of the present invention, it should be understood that this invention is not limited to certain described methods and experimental conditions, since such methods and conditions may vary. It should also be understood that the terminology used herein is only intended to describe particular embodiments and is not limiting, as the scope of the present invention will only be limited by the appended claims.

[0036] Unless otherwise indicated, all technical and scientific terms used herein have the meaning generally understood by a person skilled in the art to which this invention belongs. The term "approximately" in relation to a given numerical value means that the indicated value may differ from the given value by no more than 1%. For example, the expression "about 100" includes 99 and 101, as well as all values in between (eg, 99.1, 99.2, 99.3, 99.4, etc.).

[0037] While 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 described below. 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 expressed on T cells as part of the multimolecular T cell receptor (TCR) and consisting of a homodimer or heterodimer formed by association of two of the four receptor chains: CD3-epsilon, CD3-delta, CD3-zeta and CD3-gamma. Human CD3 epsilon contains the amino acid sequence shown in SEQ ID NO: 1; human CD3 delta contains the amino acid sequence shown in SEQ ID NO: 2; human CD3 zeta contains the amino acid sequence shown in SEQ ID NO: 3; and CD3-gamma contains the amino acid sequence shown 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 corresponding protein, polypeptide, or protein fragment, unless explicitly stated to be of a non-human species. Thus, the expression "CD3" means human CD3, unless it is indicated that it belongs to a species other than human, for example, "mouse CD3", "monkey CD3", etc.

[0040] "CD3-binding antigen-binding domain", "CD3-binding antibody", or "anti-CD3 antibody" includes antibodies and antigen-binding fragments thereof that specifically recognize one subunit of CD3 (e.g., epsilon, delta, gamma, or zeta), as well as antibodies and antigen-binding fragments thereof that specifically recognize a dimeric complex of two CD3 subunits (eg, CD3 gamma/epsilon, delta/epsilon, and zeta/zeta dimers). The antibodies and antigen-binding fragments of the present invention can bind soluble CD3 and/or CD3 expressed on the cell surface. Soluble CD3 includes native CD3 proteins as well as recombinant variants of CD3 proteins, such as, for example, monomeric and dimeric CD3 constructs lacking a transmembrane domain or otherwise not associated with the cell membrane.

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

[0042] "CD20-binding antigen-binding domain", "CD20-binding antibody", or "anti-CD20 antibody" includes antibodies and antigen-binding fragments thereof that specifically recognize CD20.

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

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

[0045] The expression "PSMA" refers to the prostate-specific membrane antigen, also known as folate hydrolase 1 (FOLH1). PSMA is an integral, non-shedding glycoprotein that is expressed at high levels in prostate epithelial cells and is a surface marker for prostate cancer. The amino acid sequence of human PSMA is shown in SEQ ID NO: 7.

[0046] "PSMA-binding antigen-binding domain", "PSMA-binding antibody", or "anti-PSMA antibody" includes antibodies and antigen-binding fragments thereof that specifically recognize PSMA.

[0047] The expression "MUC16" refers to mucin 16. MUC16 is a highly glycosylated integral membrane glycoprotein with a single transmembrane domain that is expressed at high levels in ovarian cancer. The amino acid sequence of human MUC16 is shown in SEQ ID NO: 8.

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

[0049] The expression "STEAP2" refers to six-domain transmembrane prostate epithelial antigen 2. STEAP2 is an integral six-stranded transmembrane protein that is expressed at high levels in prostate epithelial cells and is a surface marker of prostate cancer cells. STEAP2 is a 490 amino acid protein encoded by the STEAP2 gene located in humans in the chromosomal region 7q21. The amino acid sequence of human STEAP2 is shown in SEQ ID NO: 9.

[0050] "STEAP2 binding antigen-binding domain", "STEAP2-binding antibody", or "anti-STEAP2 antibody" includes antibodies and antigen-binding fragments thereof that specifically recognize STEAP2.

[0051] The term "therapeutic protein" includes any polypeptide, including antibodies and antigen-binding fragments thereof, as well as bispecific antibodies and antigen-binding fragments thereof, that are used to prevent, treat, or alleviate 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 containing at least one complementarity determining region (CDR) that specifically binds to or interacts with a particular antigen (e.g., CD20, BCMA, PSMA, MUC16, STEAP2, or CD3) . The term "antibody" includes immunoglobulin molecules containing four polypeptide chains - two heavy (H) chains and two light (L) chains - connected by disulfide bonds, as well as their multimers (eg, IgM). The term "antibody" also includes immunoglobulin molecules composed of four polypeptide chains—two heavy (H) chains and two light (L) chains—linked by disulfide bonds. Each heavy chain contains a heavy chain variable region (abbreviated herein as HCVR or V _H ) and a heavy chain constant region. The heavy chain constant region contains three domains, C _H 1 , C _H 2 and C _H 3. Each light chain contains a light chain variable region (abbreviated herein as LCVR or V _L ) and a light chain constant region. The light chain constant region contains one domain (C _L 1). The V _H and V _L regions can be further subdivided into regions of hypervariability called complementarity determining regions (CDRs) that alternate with more conserved regions called framework regions (FRs). Each V _H and V _L consists of three CDRs and four FRs, arranged from amino to carboxy in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In various embodiments, the FR antibody (or antigen-binding portion thereof) may be identical to human germline sequences, or may be naturally or artificially modified. The consensus amino acid sequence can be determined based on a comparative analysis of two or more CDRs. The term "antibody" includes "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 produced, synthetic or genetically modified polypeptide or glycoprotein that specifically binds an antigen to form a complex. Antigen-binding fragments of an antibody can be obtained, for example, from full-length antibody molecules using any suitable standard technique, such as proteolytic cleavage or recombinant genetic engineering methods, including modification and expression of DNA encoding variable and optionally constant domains of antibodies. Such DNA is known and/or readily available from, for example, commercial sources, DNA libraries (including, for example, antibody phage libraries), or can be synthesized. The DNA can be sequenced and modified chemically or by molecular biology techniques, for example, to place one or more variable and/or constant domains in a suitable configuration, or to introduce codons, create cysteine residues, modify, add or remove amino acids, etc.

[0055] The expression "bispecific antigennegative molecule" refers to a protein, polypeptide or molecular complex containing at least a first antigennegative domain and a second antigennegative domain. Each antigen-binding domain in a bispecific antigen-binding molecule contains at least one CDR, which, alone or in combination with one or more additional CDRs and/or FRs, 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 molecules (scFv); (vi) dAb fragments; and (vii) minimal recognition units consisting of amino acid residues that mimic the hypervariable region of an antibody (e.g., an isolated complementarity determining region (CDR), e.g., a CDR3 peptide), or a FR3-CDR3-FR4 conformationally restricted peptide. Other genetically modified molecules such as domain specific antibodies, single domain antibodies, domain deleted antibodies, chimeric antibodies, CDR grafted antibodies, diabodies, tribodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, divalent nanobodies, etc. ), small modular protein (SMIP) immunopharmaceuticals, and shark IgNAR variable domains are also covered by the expression "antigen binding fragment".

[0057] An antigen binding fragment of an antibody typically contains at least one variable domain. The variable domain may be of any size or amino acid composition, and typically contains at least one CDR that is directly adjacent to or in reading frame with one or more framework sequences. In antigen-binding fragments having a V _H domain associated with a V _L domain, the V _H and V _L domains may be arranged relative to each other in any suitable order. For example, the variable region may be dimeric and contain V _H -V _H , V _H -V _L or V _L -V _L dimers. Alternatively, the antigen-binding fragment of an antibody may contain a V _H or V _L monomeric domain.

[0058] In certain embodiments, an antigen-binding fragment of an antibody may comprise at least one variable domain covalently linked to at least one constant domain. Non-limiting illustrative configurations of variable and constant domains that may be contained in an antigen-binding fragment of an antibody of the present invention include: (i) V _{H -CH} 1; (ii) V _H -C _H 2; (iii) V _H -C _H 3; (iv) V _H -C _H 1 -C _H 2; (v) V _H -C _H 1 -C _H 2 -C _H 3; (vi) V _H -C _H 2 -C _H 3; (vii) V _H -C _L ; (viii) V _L -C _H 1; (ix) V _{L -CH} 2; (x) V _L -C _H 3; (xi) V _L -C _H 1 -C _H 2; (xii) V _L -C _H 1 -C _H 2 -C _H 3; (xiii) V _L -C _H 2 -C _H 3; and (xiv) V _L -C _L . In any configuration of the variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains may be either directly linked to each other, or may be linked by a complete or partial hinge or linker region. The hinge region may be at least 2 (eg, 5, 10, 15, 20, 40, 60 or more) amino acids that result in a flexible or semi-flexible bond between adjacent variable and/or constant domains in a single polypeptide molecule. In addition, an antigen-binding fragment of an antibody of the present invention may comprise a homodimer or heterodimer (or other multimer) in any of the variable and constant domain configurations listed above, in non-covalent association with each other and/or with one or more V _H or V monomeric domains. _L (eg via disulfide(s) bond(s)).

[0059] As with full-length antibody molecules, antigen-binding fragments can be monospecific or multispecific (eg, bispecific). A multispecific antigen-binding fragment of an antibody typically contains at least two different variable domains, each capable of specifically binding to its own antigen or to a different epitope on the same antigen. Any multispecific antibody format, including exemplary bispecific antibody formats disclosed herein, can be adapted for use in the context of an antigen-binding fragment of an antibody of the present invention using standard techniques available in the art.

[0060] Antibodies of the present invention can perform their function through complement-dependent cytotoxicity (CDC) or antibody-dependent cell-mediated cytotoxicity (ADCC). "Complement dependent cytotoxicity" (CDC) refers to the lysis of antigen-expressing cells by an antibody of the invention in the presence of complement. "Antibody-dependent cell-mediated cytotoxicity" (ADCC) refers to a cell-mediated response in which non-specific cytotoxic cells expressing Fc receptors (FcRs) (e.g., natural killer (NK) cells, neutrophils, and macrophages) recognize bound antibody on the target cell and thereby leading to its lysis. CDC and ADCC can be measured using assays that are well known and available in the art. (See, for example, US Pat. Nos. 5,500,362 and 5,821,337, and Clynes et al. (1998) Proc. Natl. Acad. Sci. (USA) 95:652-656). The constant region of an antibody is important for the ability of an antibody to fix complement and mediate cell-dependent cytotoxicity. Thus, an antibody isotype can be selected based on whether it is desirable for a given antibody to mediate cytotoxicity. The antibodies of the present invention may include a human IgG heavy chain. In various embodiments, the heavy chain may be of the isotype IgG1, IgG2, IgG3, or IgG4.

[0061] In certain embodiments, 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 germline immunoglobulin sequences. Human antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by in vitro random or site-directed mutagenesis or in vivo somatic mutation), for example, in CDRs and, in particular, in CDR3. However, the term "human antibody" 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 human framework sequences.

[0062] The antibodies of the invention may, in some embodiments, be recombinant human antibodies. The term "recombinant human antibody" is intended to include all human antibodies produced, expressed, generated or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell (described further below), antibodies isolated from recombinant, combinatorial library of human antibodies (described further below), antibodies isolated from an animal (e.g. mouse), transgenic for human immunoglobulin genes (see, e.g., Taylor et al. (1992) Nucl. Acids Res. 20:6287- 6295), or antibodies produced, expressed, constructed or isolated by any other method that involves splicing human immunoglobulin gene sequences to produce other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or, in the case of an animal transgenic for human Ig sequences, somatic in vivo mutagenesis) and thus the amino acid sequences of the V _H and V _L regions of the recombinant antibodies are sequences which, although derived from and related to human germline V _H and V _L sequences, cannot naturally exist in the in vivo human antibody germline repertoire.

[0063] Human antibodies can exist in two forms, which are associated with hinge differences. In one form, the immunoglobulin molecule contains a stable four chain construct of approximately 150-160 kDa in which the dimers are held together by an interchain disulfide bond between the heavy chains. In the second form, the dimers are not linked by interchain disulfide bonds, and a molecule of approximately 75-80 kDa is formed, consisting of covalently linked light and heavy chains (semi-antibody). These forms are extremely difficult to separate, even after affine purification.

[0064] The frequency of appearance of the second form in various intact IgG isotypes is due, but not limited to, structural differences associated with the isotype of the hinge region of the antibody. A single amino acid substitution at the human IgG4 hinge region can significantly reduce the incidence of the second form (Angal et al. (1993) Molecular Immunology 30:105) to levels normally seen with the human IgG1 hinge. The present invention encompasses antibodies having one or more mutations in the hinge, C _H 2 or C _H 3 regions, which may be desirable, for example during manufacture, to improve the yield of the desired antibody form.

[0065] Antibodies of the invention may be isolated antibodies. "Isolated antibody" in the context of this document means an antibody that has been identified and separated and/or extracted from at least one component of its natural environment. For example, an antibody that has been isolated or recovered from at least one component of an organism, or from a tissue or cell in which the antibody naturally exists or is naturally produced, is an "isolated antibody" in the context of the present invention. An isolated antibody also includes an antibody in situ in a recombinant cell. Isolated antibodies are antibodies that have been subjected to at least one purification or isolation step. In certain embodiments, the isolated antibody may be substantially free of other cellular material and/or chemicals.

[0066] The antibodies discussed herein may contain one or more amino acid substitutions, insertions and/or deletions in the framework regions and/or CDR regions of the heavy and light chain variable domains compared to the corresponding germline sequences from which these were derived. antibodies. Such mutations can be readily identified by comparing the amino acid sequences disclosed herein with germline sequences available, for example, from public antibody sequence databases. The present invention includes antibodies and antigen-binding fragments thereof, derived from any of the amino acid sequences disclosed herein, in which one or more amino acids in one or more framework regions and/or CDR regions are mutated into the corresponding germline sequence residue(s), from which the antibody was derived, or to the corresponding residue(s) of another human germline sequence, or to a conservative amino acid substitution of the corresponding germline residue(s) (such sequence changes are collectively referred to herein as "germline mutations"). One skilled in the art, starting from the heavy and light chain variable region sequences disclosed herein, can readily generate a plurality of antibodies and antigen-binding fragments containing one or more individual germline mutations, or combinations thereof. In certain embodiments, all framework and/or CDR residues in the V _H and/or V _L domains are mutated back to residues contained in the original germline sequence from which the antibody was derived. In other embodiments, only some residues are mutated back to the original germline sequence, such as only mutated residues contained in the first 8 amino acids of FR1 or the last 8 amino acids of FR4, or only mutated residues contained in CDR1, CDR2, or CDR3. In other embodiments, one or more framework residues and/or CDR residues are mutated to the corresponding residue(s) of a different germline sequence (i.e., a germline sequence that differs from the germline sequence from which the antibody was originally derived). In addition, the antibodies of the present invention may contain any combination of two or more germline mutations within the framework regions and/or CDR regions, for example, in which some individual residues are mutated to the corresponding residue of a particular germline sequence, while some other residues that differ from the original germline sequence, are retained or mutated into the corresponding residue of a different germline sequence. Once generated, antibodies and antigen-binding fragments containing one or more germline mutations can be readily tested for one or more desired properties, such as improved binding specificity, increased binding affinity, improved or enhanced antagonistic or agonistic biological properties (as appropriate) , reduced immunogenicity, etc. Antibodies and antigen-binding fragments obtained in this General way, are covered by the present invention.

[0067] The present invention also includes antibodies containing variants of any of the HCVR, LCVR and/or CDR amino acid sequences disclosed herein having one or more conservative substitutions. For example, the present invention includes antibodies having the amino acid sequences HCVR, LCVR, and/or CDR with, for example, 10 or less, 8 or less, 6 or less, 4 or less, etc. conservative amino acid substitutions relative to any of the HCVR, LCVR and/or CDR amino acid sequences provided herein, anti-CD3 antibodies disclosed in WO 2014/047231 or WO 2017/053856, anti-CD20 x anti-CD3 bispecific antibodies disclosed in WO 2014/047231, anti-PSMA or anti-PSMA x anti-CD3 antibodies disclosed in WO 2017/023761, anti-MUC16 or anti-MUC16 x anti-CD3 antibodies disclosed in WO 2018/067331, anti-STEAP2 or anti- STEAP2 x anti-CD3 antibodies disclosed in WO 2018/058001, or anti-BCMA or anti-BCMA x anti-CD3 antibodies disclosed in US 62/700596 (filed July 19, 2018), each of these sources is included in this document through a link.

[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. One antigen may have more than one epitope. Thus, different antibodies can bind to different regions on an antigen and can have different biological effects. Epitopes may be conformational or linear. A conformational epitope is formed by space-contiguous amino acids from different segments of a linear polypeptide chain. A linear epitope is an epitope formed by adjacent amino acid residues in a polypeptide chain. In certain circumstances, an epitope may include fragments of saccharides, phosphoryl groups, or sulfonyl groups on an antigen.

[0069] The term "substantially identical" or "substantially identical" in relation to a nucleic acid or fragment thereof means that, when optimally aligned with the corresponding nucleotide insertions or deletions with another nucleic acid (or its complementary strand), there is an identity of nucleotide bases of a nucleotide at least about 95%, and more preferably at least about 96%, 97%, 98%, or 99%, as measured by any well-known sequence identity algorithm such as FASTA, BLAST, or GAP, as discussed below. A nucleic acid molecule substantially identical to the reference nucleic acid molecule may, in certain instances, encode a polypeptide having the same or substantially similar amino acid sequence as the polypeptide encoded by said reference nucleic acid molecule.

[0070] When applied to polypeptides, the term "substantially similar" or "substantially similar" means that two peptide sequences, when optimally aligned, for example, using the GAP or BESTFIT programs using default gap opening penalties, have at least 95 % sequence identity, even more preferably at least 98% or 99% sequence identity. Preferably, positions of residues that are not identical are distinguished by conservative amino acid substitutions. A "conservative amino acid substitution" is a substitution in which an amino acid residue is replaced with another amino acid residue having a side chain (R group) with similar chemical properties (eg, charge or hydrophobicity). In general, a conservative amino acid substitution does not substantially change the functional properties of the protein. In cases where two or more amino acid sequences differ from each other by conservative substitutions, the percent identity or degree of sequence similarity may be adjusted upwards 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) main side chains: lysine, arginine and histidine; (6) acidic side chains: aspartate and glutamate, and (7) sulfur-containing side chains - cysteine and methionine. Preferred groups of conservative amino acid substitutions are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamate-aspartate, and asparagine-glutamine. Alternatively, a conservative substitution is any change that has a positive value in the PAM250 log-likelihood matrix disclosed 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, also referred to as sequence identity, is commonly measured using sequence analysis software. Protein analysis software selects similar sequences by applying 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 that can be used with default parameters to determine homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species, or between a wild-type protein and its mutein. See, for example, GCG version 6.1. Polypeptide sequences can also be compared using FASTA with default or recommended parameters - programs in GCG version 6.1. FASTA (eg, FASTA2 and FASTA3) provides alignments and percent sequence identity of regions of best overlap between a given and a found sequence (see Pearson (2000) above). Another preferred algorithm for comparing a sequence according to the invention with a database containing a large number of sequences from different organisms is the BLAST computer program, especially BLASTP or TBLASTN, using default 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 of which is incorporated herein by reference.

Dosing Strategies and Administration Schedules

[0072] Dosing strategies have been developed that provide administration regimens to reduce the incidence or severity, or both, of cytokine release syndrome (CRS) or infusion response (IR) by administering a therapeutic protein to a patient for various therapies. In certain embodiments of the present invention, these strategies include multiple doses of a therapeutic protein or antigen-binding molecule (eg, antibody or bispecific antibody) that can be administered to a subject over a period of time, thereby providing a regimen. Methods according to this aspect of the invention include sequential administration of multiple doses of a therapeutic protein according to the invention to a subject. "Sequential administration" means that each dose of a therapeutic protein is administered to a subject at a separate point in time, eg, on different days at a predetermined interval (eg, several hours, days, weeks, or months). The present invention includes methods comprising sequentially administering to a patient a fractional primary dose of a therapeutic protein, followed by a fractional secondary dose of a therapeutic protein, optionally followed by a fractional tertiary dose of a therapeutic protein, followed by single doses of the maximum weekly dose of therapeutic protein. The present regimens allow for higher doses of therapeutic protein, which are desirable to enhance therapeutic efficacy, but without the negative effects associated with CRS or IR. Without wishing to be bound by any particular theory, it is believed that the present regimens provide for the priming of an immune response to administration of a therapeutic protein, minimizing the incidence and severity of CRS and IR in the early stages of the regimen, which then allows higher doses of therapeutic proteins to be administered later in the regimen. subsequent stages of the treatment regimen without significant adverse events associated with CRS or IR.

[0073] An exemplary dosing regimen includes: (i) administering portions of a primary dose of a therapeutic protein at week 1 (W1) of a dosing regimen wherein said primary dose is not more than 1 mg of therapeutic protein, the first portion of the dose (F1D1) is 50% of the primary dose, and it is administered to the subject on day 1 of week 1 and the second dose portion (F2D1) is 50% of the total primary dose and is administered to the subject no later than 96 hours after administration of F1D1; (ii) administering portions of a secondary dose of the therapeutic protein at week 2 (W2) of a dosing regimen where said secondary dose is no more than half of the maximum weekly dose of the therapeutic protein, the first portion of the dose (F1D2) being 50% of the secondary dose, the second portion of the dose (F2D2) is 50% of the secondary dose, and F1D2 and F2D2 are administered to the subject 96 hours apart on week 2 of the dosing regimen; and (iii) administering to the subject the specified maximum weekly dose of the therapeutic protein as a single dose in the subsequent week (Ws) of the dosing regimen.

[0074] Another exemplary administration schedule includes: (i) administering portions of the primary dose of therapeutic protein at week 1 (W1) of the dosing regimen, where said primary dose is not more than 1 mg of therapeutic protein, the first portion of the dose (F1D1) is 50% of the primary dose and is administered to the subject on day 1 of week 1 and the second portion of the dose (F2D1) is 50% of the total primary dose and is administered to the subject no later than 96 hours after administration of F1D1; (ii) administering portions of a secondary dose of the therapeutic protein at week 2 (W2) of a dosing regimen where said secondary dose is no more than half of the maximum weekly dose of the therapeutic protein, the first portion of the dose (F1D2) being 50% of the secondary dose, the second portion of the dose (F2D2) is 50% of the secondary dose, and F1D2 and F2D2 are administered to the subject 96 hours apart on week 2 of the dosing regimen; (iii) administering portions of a tertiary dose of the therapeutic protein at week 1 (W3) of the dosing regimen, wherein said tertiary dose is at least half of the specified maximum weekly dose of the therapeutic protein and not more than the specified maximum weekly dose of the therapeutic protein, the first dose portion (F1D3) is 50 % of the tertiary dose, the second portion of the dose (F2D3) is 50% of the tertiary dose, and F1D3 and F2D3 are administered to the subject 96 hours apart in week 1 of the dosing regimen; and (iv) administering to the subject the specified maximum weekly dose of the therapeutic protein as a single dose in the subsequent week (Ws) of the dosing regimen.

[0075] In various embodiments, the implementation of the primary dose of therapeutic protein may be from 0.1 mg to 10 mg or more. In some cases, the initial 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 initial 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 10 mg. In various embodiments, the implementation of the primary dose of therapeutic protein includes a range that includes any of the above values as the upper or lower end of the range (for example, 1-5 mg).

[0076] In various embodiments, the maximum weekly dose of therapeutic protein is from 5 mg to 320 mg. In some embodiments, the maximum weekly dose of the therapeutic protein is greater than 5 mg up to 320 mg. In some cases, the maximum weekly dose of a 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 -160mg, 160-320mg, 5mg, 6mg, 7mg, 8mg, 12mg, 18mg, 27mg, 40mg, 80mg, 160mg or 320mg. 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, 175, 180, 185, 190, 195, 200, 205, 210, 215 . In various embodiments, the implementation of the maximum weekly dose of therapeutic protein includes a range including any of the above values as the upper or lower end of the range (for example, 200-300 mg).

[0077] In various embodiments, the secondary dose is 50% of the maximum weekly dose of the therapeutic protein. In some cases, the secondary dose is 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 therapeutic protein. In some cases, the secondary dose of the therapeutic protein includes a range as a percentage of the maximum weekly dose, including any of the above values as the upper or lower end of the range (for example, 35-50%).

[0078] In various embodiments, the implementation of the tertiary dose is 50% of the maximum weekly dose of therapeutic protein. In some cases, the tertiary dose is 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% or 90% of the maximum weekly dose of therapeutic protein. In some cases, the tertiary dose of the therapeutic protein includes a range as a percentage of the maximum weekly dose, including any of the above values as the upper or lower end of the range (eg, 50-75%).

[0079] In various embodiments, each of the first dose portion and the second dose portion of the primary, secondary, and/or tertiary doses is 50% of the dose. In some cases, the first dose portion and the second dose portion of the primary, secondary, and/or tertiary doses constitute different percentages (adding up to 100%) of the total dose. For example, the first portion of the dose may be 45% of the dose, and the second portion of the dose may be 55% of the dose. Alternatively, the first portion of the dose may be 55% of the dose and the second portion of the dose may be 45% of the dose. In various embodiments, the implementation of the first and second parts 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 implementation of the primary, secondary and/or tertiary doses (D1, D2 and/or D3) can be divided into two or more parts. The various options for dividing doses into two parts have been discussed above. However, in some cases, the doses are divided into 3, 4 or 5 parts. For example, the primary dose may be divided into 5 portions, each representing 20% of the total primary dose, and each portion of the dose (F1D1, F2D1, F3D1, F4D1, and F5D1) may be administered to the subject for five consecutive days in the first week of the dosing regimen. . In other cases, for each part of the dose, the percentage of the total dose (for example, the primary dose) may be different. For example, if the primary dose is divided into 3 portions, the first portion of the dose (F1D1) may include 30% of the total primary dose, the second portion of the dose (F2D1) may include 30% of the total primary dose, and the third portion of the dose (F3D1) may include the remaining 40% total primary dose. Other combinations of percentages and number of fractional doses that add up to 100% of the total dose are expressly provided herein.

[0081] In one illustrative embodiment of the dosing regimen, the primary dose is 1 mg, each of the first dose portion (F1D1) and the second dose portion (F2D1) is 500 μg, the secondary dose is 6 mg, each of the first dose portion (F1D2) and the second dose portion (F2D2) is 3 mg, the tertiary dose is 12 mg, the first dose portion (F1D3) and the second dose portion (F2D3) are each 6 mg, and the maximum weekly dose is 12 mg of therapeutic protein.

[0082] In one illustrative embodiment of the dosing regimen, the primary dose is 1 mg, each of the first dose portion (F1D1) and the second dose portion (F2D1) is 500 μg, the secondary dose is 9 mg, each of the first dose portion (F1D2) and the second portion of the dose (F2D2) is 4.5 mg, the tertiary dose is 18 mg, each of the first portion of the dose (F1D3) and the second portion of the dose (F2D3) is 9 mg, and the maximum weekly dose is 18 mg of therapeutic protein.

[0083] In one illustrative embodiment of the dosing regimen, the primary dose is 1 mg, each of the first dose portion (F1D1) and the second dose portion (F2D1) is 500 μg, the secondary dose is 13.5 mg, each of the first dose portion (F1D2 ) and the second dose portion (F2D2) is 6.75 mg, the tertiary dose is 27 mg, the first dose portion (F1D3) and the second dose portion (F2D3) are each 13.5 mg, and the maximum weekly dose is 27 mg of therapeutic protein .

[0084] In one illustrative embodiment of the dosing regimen, the primary dose is 1 mg, each of the first dose portion (F1D1) and the second dose portion (F2D1) is 500 μg, the secondary dose is 20 mg, each of the first dose portion (F1D2) and the second dose installment (F2D2) is 10 mg, the tertiary dose is 40 mg, the first dose installment (F1D3) and the second dose installment (F2D3) are each 20 mg, and the maximum weekly dose is 40 mg of therapeutic protein.

[0085] In one illustrative embodiment of the dosing regimen, the primary dose is 1 mg, each of the first dose portion (F1D1) and the second dose portion (F2D1) is 500 μg, the secondary dose is 20 mg, each of the first dose portion (F1D2) and the second dose installment (F2D2) is 10 mg, the tertiary dose is 60 mg, the first dose installment (F1D3) and the second dose installment (F2D3) are each 30 mg, and the maximum weekly dose is 80 mg of therapeutic protein.

[0086] In one illustrative embodiment of the dosing regimen, the primary dose is 1 mg, each of the first dose portion (F1D1) and the second dose portion (F2D1) is 500 μg, the secondary dose is 20 mg, each of the first dose portion (F1D2) and the second dose portion (F2D2) is 10 mg, the tertiary dose is 80 mg, each of the first portion (F1D3) and the second portion (F2D3) is 40 mg, and the maximum weekly dose is 160 mg of therapeutic protein.

[0087] In one illustrative embodiment of the dosing regimen, the primary dose is 1 mg, each of the first dose portion (F1D1) and the second dose portion (F2D1) is 500 μg, the secondary dose is 20 mg, each of the first dose portion (F1D2) and the second portion of the dose (F2D2) is 10 mg, the tertiary dose is 120 mg, each of the first portion of the dose (F1D3) and the second portion of the dose (F2D3) is 60 mg, and the maximum weekly dose is 240 mg of therapeutic protein.

[0088] In one illustrative embodiment of the dosing regimen, the primary dose is 1 mg, each of the first dose portion (F1D1) and the second dose portion (F2D1) is 500 μg, the secondary dose is 20 mg, each of the first dose portion (F1D2) and the second dose portion (F2D2) is 10 mg, the tertiary dose is 160 mg, each of the first portion (F1D3) and the second portion (F2D3) is 80 mg, and the maximum weekly dose is 320 mg of therapeutic protein.

[0089] In one illustrative embodiment of the dosing regimen, the primary dose (D1) is 1 mg, each of the first dose portion (F1D1) and the second dose portion (F2D2) of the primary dose is 500 μg, the secondary dose (D2) is 3 mg, each of the first dose part (F1D2) and the second dose part (F2D2) of the secondary dose is 1.5 mg and the maximum weekly dose is 3 mg of therapeutic protein.

[0090] In one illustrative embodiment of the dosing regimen, the primary dose (D1) is 3 mg, each of the first dose portion (F1D1) and the second dose portion (F2D2) of the primary dose is 1.5 mg, the secondary dose (D2) is 9 mg , each of the first dose part (F1D2) and the second dose part (F2D2) of the secondary dose is 4.5 mg, and the maximum weekly dose is 9 mg of therapeutic protein.

[0091] In one illustrative embodiment of the dosing regimen, the primary dose (D1) is 5 mg, each of the first dose portion (F1D1) and the second dose portion (F2D2) of the primary dose is 2.5 mg, the secondary dose (D2) is 15 mg , each of the first dose part (F1D2) and the second dose part (F2D2) of the secondary dose is 7.5 mg, and the maximum weekly dose is 15 mg of therapeutic protein.

[0092] In one illustrative embodiment of the dosing regimen, the primary dose (D1) is 10 mg, each of the first dose portion (F1D1) and the second dose portion (F2D2) of the primary dose is 5 mg, the secondary dose (D2) is 30 mg, each of the first dose part (F1D2) and the second dose part (F2D2) of the secondary dose is 15 mg and the maximum weekly dose is 30 mg of therapeutic protein.

[0093] In various embodiments, the therapeutic protein is administered at a dose that maintains a serum concentration of at least about 2000 µg/L after administration of the maximum weekly dose throughout the dosing regimen. In some cases, the therapeutic protein is administered at a dose that maintains 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 the introduction of the maximum weekly dose throughout the dosing regimen.

[0094] In various embodiments, the therapeutic protein is administered at a dose that maintains an average serum concentration of at least about 2600 μg/L after administration of the maximum weekly dose throughout the dosing regimen. In some instances, the therapeutic protein is administered at a dose that maintains mean serum levels of at least about 2000, 2100, 2200, 2300, 2400, 2500, 2700, 2800, 2900, 3000, 3100, 3200, 3300, 3400, 3500, 3600, 3700, 3800, 3900 or 4000 mcg/l after the maximum weekly dose during the dosing regimen.

[0095] In various embodiments, the subsequent week (Ws) of the dosing schedule 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 (W11), 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 35 (W34 ), week 35 (W35) or week 36 (W36).

[0096] In various embodiments, the second fractional dose in any given week of the dosing regimen is administered no later than 24, 36, 48, 60, 72, 84, or 96 hours after the first fractional 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, 23, 24, 25, 26 or more weeks during the 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 for a number of weeks in a range including any of the above values as 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) every other week for up to 24 weeks or 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 the twice-weekly, three-times-weekly, four-times-weekly, or five-times-weekly (maintenance phase) phase of the dosing regimen, which may follow the completion of the weekly phase of the dosing regimen (ie. e. either weekly dosing of the maximum weekly dose, or divided dosing of primary, secondary and (optionally) tertiary doses). In some instances, the maximum weekly dose of a therapeutic protein is administered to a subject as a single dose (maintenance dose) once every two weeks, once every three weeks, or once every four weeks for a number of weeks in the range including any of the above values as the upper or lower end of the range (for example, 24-86 weeks).

[0099] In one illustrative embodiment of the dosing regimen, the primary dose is 1 mg, each of the first dose portion (F1D1) and the second dose portion (F2D1) is 500 μg, the secondary dose is 20 mg, each of the first dose portion (F1D2) and the second dose portion (F2D2) is 10 mg, the tertiary dose is 80 mg, each of the first dose portion (F1D3) and the second dose portion (F2D3) is 40 mg, and the maximum weekly dose is 160 mg of therapeutic protein, where the tertiary dose is administered in as a single dose (i.e., 80 mg) weekly (1p/week) during weeks 4 to 12 of the dosing regimen, and the maximum weekly dose is administered as a single dose (i.e., 160 mg) once every two weeks ( 1p/2wk), starting from week 14 of the dosing regimen.

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

[0101] In one illustrative embodiment of the dosing regimen, the primary dose is 1 mg, each of the first dose portion (F1D1) and the second dose portion (F2D1) is 500 μg, the secondary dose is 20 mg, each of the first dose portion (F1D2) and the second dose portion (F2D2) is 10 mg, the tertiary dose is 160 mg, each of the first dose portion (F1D3) and the second dose portion (F2D3) is 80 mg, and the maximum weekly dose is 320 mg of therapeutic protein, where the tertiary dose is administered in as a single dose (i.e. 160 mg) weekly (1p/week) during weeks 4 to 12 of the dosing regimen, and the maximum weekly dose is administered as a single dose (i.e. 320 mg) once every two weeks ( 1p/2wk), starting at week 14 of the dosing regimen.

[0102] In some instances, the above dosing regimen is for use in methods of treating aggressive lymphoma (eg, mantle cell lymphoma or marginal zone lymphoma). In some instances, the above dosing regimen is for use in methods of treating follicular lymphoma (eg, steps 1-3a). In some cases, the above dosing regimen is indicated for the treatment of diffuse large B-cell lymphoma (including relapsed or refractory DLBCL, for example, in patients who have failed previous CAR-T therapy).

[0103] In various embodiments, each dose or fractional dose of a therapeutic protein is administered to a subject over 1-4, 1-5, or 1-6 hours (eg, via infusion). In some cases, the dose or sub-dose is administered over 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 or more hours. In some cases, each dose or sub-dose of a therapeutic protein is administered to a subject over a time range including any of the above values as the upper or lower end of the range (eg, 1-8 hours). In various embodiments, 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 pre-treatment or in combination with a therapeutic protein. In some instances, a steroid, such as dexamethasone, is administered to the patient as a pre-treatment prior to the administration of primary split doses and secondary split doses, and optionally prior to tertiary split doses. In some embodiments, dexamethasone is administered to the patient approximately one to three hours prior to the first portion of the dose (F1D1). In some embodiments, the implementation of the dose of dexamethasone is administered by intravenous infusion. In some cases, a cytokine antagonist such as an anti-IL-6 antibody or an anti-IL-6R antibody is administered in combination with a therapeutic protein during primary, secondary, and optionally tertiary dosing. In some instances, a patient is administered an anti-CD20 monospecific antibody (eg, rituximab) as a pre-treatment prior to primary fractional doses and optionally secondary fractional doses, and optionally prior to tertiary fractional doses. These administrations of the second agent as pre-treatment or as a combination are terminated, in one embodiment, with the administration of the maximum weekly dose as a single dose, which begins, for example, at week 3, 4, 5, or 6 of the dosing regimen, so as not to artificially stifle cytokine activity and not thereby interfere with the therapeutic effects of the therapeutic protein.

[0105] The terms "primary dose", "secondary dose" and "tertiary dose" refer to the time sequence of administration of the antigen-binding molecule of the invention. Thus, the "primary dose" is the dose that is administered at the beginning of the treatment regimen; "secondary dose" is a dose that is administered after the primary dose (including all dose portions for the primary dose); and "tertiary doses" are doses that are administered after the secondary dose (including all dose portions for the secondary dose). Primary, secondary and tertiary doses (or parts of these doses) may contain a certain amount of therapeutic protein, as discussed in this document. In some cases, the "primary dose" and "secondary dose" and 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 "initial dose". In some cases, a "secondary dose" may be referred to as an "intermediate dose". In some cases, a "tertiary dose" may be referred to as a "step-up dose".

[0106] The phrase "immediately preceding dose" in a multi-dose sequence refers to the dose of the antigen-binding molecule that is administered to the patient prior to the immediate next dose in the sequence with no intermediate doses.

Antigen binding molecules

[0107] Antibodies for use according to the present invention may be monospecific, bispecific, or multispecific. Multispecific antibodies may be specific for different epitopes of the same 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 et al ., 2004, Trends Biotechnol. 22:238-244. The antibodies and bispecific molecules of the present invention may be linked to or co-expressed with another functional molecule, such as another peptide or protein. For example, an antibody or antibody fragment may be operably linked (e.g., by chemical linkage, genetic fusion, non-covalent association, or otherwise) 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 amino acid sequences of HCVR, LCVR and/or CDRs of the antibodies described herein, anti-CD3 antibodies disclosed in WO 2014/047231 or WO 2017/053856, anti-CD20 x anti-CD3 bispecific antibodies , disclosed in WO 2014/047231, anti-PSMA or anti-PSMA x anti-CD3 antibodies disclosed in WO 2017/023761, anti-MUC16 or anti-MUC16 x anti-CD3 antibodies disclosed in WO 2018/067331, anti- STEAP2 or anti-STEAP2 x anti-CD3 antibodies disclosed in WO 2018/058001, or anti-BCMA or anti-BCMA x anti-CD3 antibodies disclosed in US 62/700596 (filed July 19, 2018), each of these sources are incorporated herein by reference.

[0109] When using the expressions "anti-CD3 antibody" (anti-CD3 antibody), "anti-CD20 antibody", "anti-PSMA antibody", "anti-MUC16 antibody", "anti-STEAP2 antibodya", "anti-BCMA antibody", etc. both monospecific antibodies and bispecific antibodies containing the appropriate antigen-binding arm (eg, CD3) are meant herein. Thus, the present invention includes bispecific antibodies in which one immunoglobulin arm binds human CD3 and the other immunoglobulin arm is specific to, for example, human CD20, human PSMA, human MUC16, human STEAP2, or human BCMA.

[0110] In certain embodiments, the CD3 binding arm binds to human CD3 and induces the activation of human T cells. In certain embodiments, the CD3 binding arm binds weakly to human CD3 and induces the activation of human T cells. In other embodiments, the CD3 binding arm binds weakly to human CD3 and induces the killing of cells expressing the tumor associated antigen in the context of a bispecific or multispecific antibody. In other embodiments, the CD3 binding arm binds or associates weakly with human and cynomolgus monkey (monkey) CD3, however, the binding interaction is not detectable by in vitro assays known in the art. In certain embodiments, the 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, OX40, 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 contains a heavy chain variable domain (HCVR) and a light chain variable domain (LCVR). In the context of a bispecific antigen-binding molecule comprising a first and a second antigen-binding domain (e.g., a bispecific antibody), the CDRs of the first antigen-binding domain may be denoted by the prefix "A1" and the CDRs of the second antigen-binding domain may be denoted by the prefix "A2". Thus, 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 a bispecific anti-CD20 x anti-CD3 antibody known as REGN1979. REGN1979 contains a CD20 binding arm containing a heavy chain containing the amino acid sequence of SEQ ID NO: 10, a CD3 binding arm containing a heavy chain containing the amino acid sequence of SEQ ID NO: 11, and a common light chain (corresponding to both the CD20 and to CD3) containing the amino acid sequence of SEQ ID NO: 12. In some cases, the mature form of the antibody may not include C-terminal lysine residues from SEQ ID NO: 10 and 11. Thus, in some cases, the binding arm to CD20 REGN1979 contains a heavy chain containing residues 1-452 of SEQ ID NO: 10, and the binding arm to CD3 REGN1979 contains a heavy chain containing residues 1-448 of SEQ ID NO: 11. REGN1979 also contains the HCVR, LCVR and CDR sequences shown in the attached sequence listing . HCVR to CD20 corresponds to SEQ ID NO: 13, HCVR to CD3 corresponds to SEQ ID NO: 14, and total LCVR corresponds to SEQ ID NO: 15. Domains to CD20 HCDR1-HCDR2-HCDR3 correspond to SEQ ID NO: 16-17-18, respectively . Domains to CD3 HCDR1-HCDR2-HCDR3 correspond to SEQ ID NO: 19-20-21, respectively. The common domains LCDR1-LCDR2-LCDR3 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 contains a human IgG heavy chain constant region. In some cases, the human IgG heavy chain constant region is of the IgG1 isotype. In some cases, the human IgG heavy chain constant region is of the IgG4 isotype. In various embodiments, the bispecific antibody comprises a chimeric hinge that reduces binding to the Fcɣ receptor compared to a wild-type hinge of the same isotype. In some instances, 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, incorporated herein by reference.

[0114] The first antigen-binding domain and the second antigen-binding domain can be directly or indirectly linked to each other to form a bispecific antigen-binding molecule according to the present invention. Alternatively, each of the first antigen-binding domain and the second antigen-binding domain can be connected to a separate multimerization domain. The association of one multimerizing domain with another multimerizing domain facilitates the association between the two antigen-binding domains, thereby allowing the formation of a bispecific antigen-binding molecule. "Multimerization domain" means any macromolecule, protein, polypeptide, peptide or amino acid that has the ability to associate with a second multimerization domain of the same or similar structure or composition. For example, the multimerizing domain may be a polypeptide containing an immunoglobulin C _H 3 domain. A non-limiting example of a multimerizing component is the Fc portion of an immunoglobulin (comprising a C _H 2 -CH ₃ domain), such as the Fc domain of an IgG selected from IgG1, IgG2, IgG3, and IgG4 isotypes, and any allotype within each isotype group.

[0115] The bispecific antigen-binding molecules of the present invention typically comprise two multimerizing domains, eg two Fc domains, each individually part of a separate antibody heavy chain. The first and second multimerizing domains may be of the same IgG isotype, eg IgG1/IgG1, 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 implementation of the multimerization domain is an Fc fragment or an amino acid sequence from 1 to about 200 amino acids in length, containing at least one cysteine residue. In other embodiments, the multimerization domain is a cysteine residue or a short cysteine-containing peptide. Other multimerizing domains include peptides or polypeptides containing or consisting of a leucine zipper, a helix-loop motif, or a supercoil motif.

[0117] Any format or technology of bispecific antibodies can be used to produce the bispecific antigen-binding molecules of the present invention. For example, an antibody or fragment thereof having a first antigen binding specificity may be operably linked (e.g., by chemical linkage, genetic fusion, non-covalent association, or otherwise) to one or more other molecular entities, such as another antibody or antibody fragment, having a second antigen-binding specificity to provide a bispecific antigen-binding molecule. Specific exemplary bispecific formats that may be used in the context of the present invention include, but are not limited to, for example, scFv or diabodies based bispecific formats, IgG-scFv fusions, double variable domain (DVD)-Ig, quadrohybridoma, bump-to-trough ”, a common light chain (for example, a common light chain with “protrusions into depressions” interaction, etc.), CrossMab, CrossFab, (SEED)body, leucine lightning, Duobody, IgG1 / IgG2, Fab (DAF) - double action lgG and Mab ² bispecific formats (for a review of the above formats, see, for example, Klein et al . 2012, mAbs 4:6, 1-11, and references cited therein).

[0118] In the context of the bispecific antigen-binding molecules of the present invention, multimerizing domains, e.g., Fc domains, may contain one or more amino acid changes (e.g., insertions, deletions, or substitutions) compared to the naturally occurring version of the wild-type Fc domain. For example, the invention includes bispecific antigen-binding molecules containing one or more modifications in the Fc domain, resulting in a modified Fc domain having a modified binding interaction (eg, increased or decreased) between Fc and FcRn. In one embodiment, the bispecific antigen-binding molecule comprises a modification in the C _H 2 or _CH 3 region, said modification increasing the affinity of the Fc domain for FcRn in an acidic environment (e.g., in an endosome where the pH is from about 5.5 to about 6 ,0). Non-limiting examples of such Fc modifications include, for example, a modification at position 250 (eg, E or Q); 250 and 428 (for example, L or 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 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 includes modification 428L (eg, M428L) and 434S (eg, N434S); modification 428L, 259I (for example, V259I) and 308F (for example, V308F); modification 433K (for example, H433K) and 434 (for example, 434Y); modification 252, 254 and 256 (for example, 252Y, 254T and 256E); modification 250Q and 428L (for example, T250Q and M428L); and modification 307 and/or 308 (eg, 308F or 308P).

[0119] The present invention also includes bispecific antigen-binding molecules comprising a first C _H 3 domain and a second C _H 3 Ig domain, wherein the first and second C _H 3 Ig domains differ from each other by at least one amino acid, and wherein the difference is at least in one amino acid reduces the binding of the bispecific antibody to protein A compared to a bispecific antibody with no difference in amino acid. In one embodiment, the first C _H 3 Ig domain binds protein A, and the second C _H 3 Ig domain contains a mutation that reduces or abolishes protein A binding, such as an H95R modification (IMGT exon numbering; H435R EU numbering). The second C _H 3 may further comprise the modification Y96F (according to IMGT; according to Y436F according to EU). See, for example, US Pat. No. 8,586,713. Other modifications that may be contained in the second _CH 3 include: D16E, L18M, N44S, K52N, V57M, and V82I (as per IMGT; D356E, L358M, N384S, K392N, V397M, and V422I according to EU) in the case of IgG1 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 (as per IMGT; Q355R, N384S, K392N, V397M, R409K, E419Q and V422I as per EU) for IgG4 antibodies.

[0120] In certain embodiments, the Fc domain may be chimeric and contain a combination of Fc sequences derived from more than one immunoglobulin isotype. For example, a chimeric Fc domain may comprise part or all of the C _H 2 sequence derived from the C _H 2 region of human IgG1, human IgG2, or human IgG4 and part or all of the C _H 3 sequence derived from human IgG1, human IgG2, or human IgG4. . The chimeric Fc 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 in combination with a "lower hinge" sequence derived from a human IgG1, human IgG2, or human IgG4 hinge region. A specific example of a chimeric Fc domain that can be included in any of the antigen-binding molecules provided herein contains, N- to C-terminus: [IgG4 C _H 1] - [IgG4 upper hinge] - [IgG2 lower hinge] - [IgG4 CH2] - [IgG4 CH3]. Another example of a chimeric Fc domain that can be included in any of the antigen-binding molecules provided herein contains, N- to C-terminus: [IgG1 C _H 1] - [IgG1 upper hinge] - [IgG2 lower hinge ] - [IgG4 CH2] - [IgG1 CH3]. These and other examples of chimeric Fc domains that can be included in any of the antigen binding molecules of the present invention are described in US Patent Application Publication No. 2014/0243504, published August 28, 2014, incorporated herein in its entirety. Chimeric Fc domains having these common structural features and variants thereof may have altered Fc receptor binding, which in turn affects Fc effector function.

Binding properties of antigen-binding molecules

[0121] The term "binding" in the context of the binding of an antibody, immunoglobulin, antibody-binding fragment, or Fc-containing protein to any, for example, predefined antigen, such as a cell surface protein or fragment thereof, usually refers to an interaction or association between at least two entities or molecular structures, such as an antibody-antigen interaction.

[0122] For example, binding affinity typically corresponds to a K _D value of about 10 ^-7 M or less, for example, about 10 ^-8 M or less, for example, about 10 ^-9 M or less, when determined, for example, using surface plasmon technology resonance (SPR) on a BIAcore 3000 instrument using the antigen as ligand and antibody, Ig, antibody binding fragment, or Fc-containing protein as analyte (or anti-ligand). Cell-based binding strategies such as fluorescence-activated cell sorting (FACS) binding assays are also commonly used, and FACS data correlate well with other methods such as competitive radioligand binding 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, an antibody or antigen-binding protein of the present invention binds to a predetermined antigen or cell surface molecule (receptor) with an affinity corresponding to a K _D value that is at least ten times less than its affinity for binding to a non-specific antigen ( e.g. BSA, casein). According to the present invention, an antibody affinity corresponding to a K _D value equal to or less than ten times less than for a non-specific antigen can be considered as non-detectable binding, however, such an antibody can be paired with a second antigen-binding arm to obtain a bispecific antibody according to the invention.

[0124] The term “K _D ” (M) refers to the equilibrium dissociation constant of a particular antibody-antigen interaction, or the equilibrium dissociation constant of the binding of an antibody or antibody binding fragment to an antigen. Between K _D and binding affinity there is an inverse relationship, so the smaller the value of K _D , the higher, that is, stronger, affinity. Thus, the terms "higher affinity" or "stronger affinity" refer to a higher ability to interact and therefore a lower K _D value, and conversely, the terms "lower affinity" or "weaker affinity" refer to to a lower reactivity and therefore a higher K _D value. In some cases, a higher binding affinity (or K _D ) of a particular molecule ( e.g. antibody) to its binding partner molecule (e.g. antigen X) compared to with the binding affinity of that molecule (e.g., an antibody) to another binding partner molecule (e.g., a Y antigen) can be expressed as a binding ratio determined by dividing the larger K _D value (lower or weaker affinity) by the smaller K _D ( higher or stronger affinity), eg expressed as 5-fold or 10-fold binding affinity, as the case may be.

[0125] The term "k _d " (c-1 or 1/c) refers to the dissociation rate constant of a particular antibody-antigen interaction, or the dissociation rate constant of an antibody or antibody binding fragment. The specified value is also referred to as the k _off value.

[0126] The term " _ka " (M-1 x c-1 or 1/M) refers to the association rate constant of a particular antibody-antigen interaction, or the association rate constant of an antibody or antibody binding fragment.

[0127] The term "K _A " (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 k _a by k _d .

[0128]The term "EC50" or "EC₅₀" refers to the half maximum effective concentration, which includes the concentration of antibody that induces a response equal to half between the initial value and the maximum after the specified exposure time. EU₅₀is essentially the concentration of the antibody at which 50% of its maximum effect is observed. In some embodiments, the EC value₅₀ equal to the concentration of the antibody according to the invention, providing half of the maximum binding to cells expressing, for example, CD3 or a tumor-associated antigen (for example, CD20), when determined, for example, using FACS binding analysis. Thus, reduced or weaker binding is observed with an increased EC value.₅₀ or half maximum effective concentration.

[0129] In one embodiment, reduced binding can be defined as an increased EC ₅₀ concentration of an antibody that binds to half the maximum number of target cells.

[0130] In yet another embodiment, the EC ₅₀ value is the concentration of an antibody of the invention that causes half-maximal depletion of target cells by T cell cytotoxic activity. Thus, increased cytotoxic activity (eg, T cell-mediated killing of tumor cells) occurs at a reduced EC ₅₀ or half maximum effective concentration.

Sequence Options

[0131] The antibodies and bispecific antigen-binding molecules of the present invention may contain one or more amino acid substitutions, insertions and/or deletions in the framework regions and/or CDR regions of the heavy and light chain variable domains compared to the corresponding germline sequences from which they were derived. separate antigen-binding domains. Such mutations can be readily identified by comparing the amino acid sequences disclosed herein with germline sequences available, for example, from public antibody sequence databases. The antigen-binding molecules of the present invention may comprise antigen-binding domains derived from any of the exemplary amino acid sequences disclosed herein, in which one or more amino acids in one or more framework regions and/or CDR regions are mutated into the corresponding germline sequence residue(s). from which the antibody was derived, or to the corresponding residue(s) of another human germline sequence, or to a conservative amino acid substitution of the corresponding germline residue(s) (such sequence changes are collectively referred to herein as "germline mutations"). One skilled in the art, starting from the heavy and light chain variable region sequences disclosed herein, can readily generate a plurality of antibodies and antigen-binding fragments containing one or more individual germline mutations, or combinations thereof. In certain embodiments, all framework and/or CDR residues in the V _H and/or V _L domains are mutated back to residues contained in the original germline sequence from which the antigen binding domain was originally derived. In other embodiments, only some residues are mutated back to the original germline sequence, such as only mutated residues contained in the first 8 amino acids of FR1 or the last 8 amino acids of FR4, or only mutated residues contained in CDR1, CDR2, or CDR3. In other embodiments, one or more framework residues and/or CDR residues are mutated to the corresponding residue(s) of a different germline sequence (i.e., a germline sequence that differs from the germline sequence from which the antigen-binding domain was originally derived). In addition, antigen-binding domains may contain any combination of two or more germline mutations within the framework regions and/or CDR regions, for example, in which some individual residues are mutated to the corresponding residue of a particular germline sequence, while some other residues that differ from of the original germline sequence are retained or mutated into the corresponding residue of another germline sequence. Once generated, antigen-binding domains containing one or more germline mutations can be readily tested for one or more desired properties, such as improved binding specificity, increased binding affinity, improved or enhanced antagonistic or agonistic biological properties (as appropriate), reduced immunogenicity, etc. Bispecific antigen-binding molecules containing one or more antigen-binding domains derived from this general principle are within the scope of the present invention.

pH dependent binding

[0132] 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, antibodies of the invention may show increased binding to, for example, a tumor antigen such as CD20 at acidic pH compared to neutral pH. The expression "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. The expression "neutral pH" means a pH from about 7.0 to about 7.4. The expression "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.

[0133] In some cases, "reduced binding ... at acidic pH compared to neutral pH" is expressed by the ratio of the K _D value of the binding of an antibody to its antigen at acidic pH to the K _D value of the binding of an antibody to its antigen at neutral pH (or vice versa ). For example, an antibody or antigen-binding fragment thereof can be considered to exhibit "reduced binding to, for example, CD20 at acidic pH compared to neutral pH" in the context of the present invention if said antibody or antigen-binding fragment thereof exhibits an acidic/neutral K _D ratio of about 3.0 or more. In certain exemplary embodiments, the acid/neutral K _D ratio for an antibody or antigen-binding fragment of the present invention may 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 more.

[0134] Antibodies with pH-dependent binding characteristics can be obtained, for example, by screening a population of antibodies for reduced (or increased) binding to a particular antigen at acidic pH compared to neutral pH. In addition, modifications to 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, in a CDR) with a histidine residue, an antibody with reduced antigen binding at acidic pH compared to neutral pH can be obtained.

Antibodies containing Fc variants

[0135] In certain embodiments, the present invention provides antibodies and bispecific antigen-binding molecules comprising an Fc domain containing one or more mutations that increase or decrease antibody binding to the FcRn receptor, e.g., at acidic pH versus neutral pH. For example, the present invention includes antibodies containing a mutation in the C _H 2 or C _H 3 region of the Fc domain, wherein the mutation(s) increases the affinity of the Fc domain for FcRn in an acidic environment (e.g., in an endosome where the pH is between about 5, 5 to about 6.0). Such mutations can lead to an increase in the serum half-life of the antibody when administered to an animal. Non-limiting examples of such Fc modifications include, for example, a modification at position 250 (eg, E or Q); 250 and 428 (for example, L or 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 includes modification 428L (eg, M428L) and 434S (eg, N434S); modification 428L, 259I (for example, V259I) and 308F (for example, V308F); modification 433K (for example, H433K) and 434 (for example, 434Y); modification 252, 254 and 256 (for example, 252Y, 254T and 256E); modification 250Q and 428L (for example, T250Q and M428L); and modification 307 and/or 308 (eg, 308F or 308P).

[0136] For example, the present invention includes antibodies and bispecific antigen-binding molecules containing an Fc domain containing 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 (eg M252Y, S254T and T256E); 428L and 434S (eg M428L and N434S); and 433K and 434F (eg H433K and N434F). All possible combinations of the aforementioned Fc domain mutations and other mutations in the antibody variable domains disclosed herein are contemplated within the scope of the present invention.

Preparation of antigen-binding domains and construction of bispecific molecules

[0137] Antigen-binding domains specific for particular antigens can be obtained using any technology for producing antibodies known in the art. Once obtained, two different antigen-binding domains specific for two different antigens (eg, CD3 and CD20) can be appropriately positioned relative to each other to obtain a bispecific antigen-binding molecule according to the present invention using standard methods. (Discussion of exemplary bispecific antibody formats that can be used to construct bispecific antigen-binding molecules of the present invention is provided elsewhere in this document). In certain embodiments, one or more individual components (eg, heavy and light chains) of the multispecific antigen-binding molecules of the invention are derived from chimeric, humanized, or fully human antibodies. Methods for making such antibodies are well known in the art. For example, one or more heavy and/or light chains of the bispecific antigen-binding molecules of the present invention can be prepared using VELOCIMMUNE™ technology. Using VELOCIMMUNE™ technology (or any other human antibody technology), chimeric antibodies with high affinity for a specific antigen (eg, CD3 or CD20) having a human variable region and a mouse constant region are initially isolated. Antibodies are characterized and selected for desired characteristics, including affinity, selectivity, epitope, etc. The mouse constant regions are replaced with the desired human constant region to create fully human heavy and/or light chains that can be incorporated into the bispecific antigen binding molecules of the present invention.

[0138] Genetically modified animals can be used to produce human bispecific antigen-binding molecules. For example, a genetically modified mouse incapable of rearranging and expressing an endogenous mouse immunoglobulin light chain variable region sequence can be used, wherein the mouse expresses only one or two human light chain variable domains encoded by human immunoglobulin sequences operably linked to the mouse kappa constant region gene. at the endogenous mouse kappa locus. Such genetically modified mice can be used to generate fully human bispecific antigen binding molecules containing two different heavy chains associated with an identical light chain containing a variable domain derived from one of two different human variable light chain gene segments. (See, for example, US 2011/0195454). Fully human refers to an antibody, or an antigen-binding fragment thereof, or an immunoglobulin domain thereof, comprising the amino acid sequence encoded by DNA derived from a human sequence, throughout the entire length of each antibody polypeptide, or an antigen-binding fragment thereof, or its immunoglobulin domain. In some cases, a fully human sequence is obtained from a protein endogenous to humans. In other cases, a fully human protein or protein sequence contains a chimeric sequence in which the sequence of each of the components is derived from a human sequence. Without wishing to be bound by any particular theory, chimeric proteins, or chimeric sequences in general, are designed to minimize the creation of immunogenic epitopes at component sequence junctions, for example, compared to any regions or domains of wild-type human immunoglobulin.

Bioequivalents

[0139] The present invention encompasses antigen-binding molecules having amino acid sequences that differ from the exemplary molecules disclosed herein but retain the ability to bind the same antigen or antigens. Such variant molecules contain one or more additions, deletions or substitutions of amino acids compared to the original sequence, but exhibit biological activity that is essentially equivalent to that of the described antibodies or bispecific antigen-binding molecules.

[0140] The present invention includes antigen-binding molecules bioequivalent to any of the exemplary antigen-binding molecules provided herein. Two antigen-binding proteins or antibodies are considered to be bioequivalent if, for example, they are pharmaceutical equivalents or pharmaceutical alternatives, the rate and extent of absorption of which do not show a significant difference when administered at the same molar dose under similar experimental conditions, both in a single dose and in multiple doses. Some antigen-binding proteins will be considered equivalents or pharmaceutical alternatives if they are equivalent in their extent of absorption, but not in their rate of absorption, and yet may be considered bioequivalent because such differences in absorption rate are intentional and reflected in the labeling, do not have essential to achieve effective drug concentrations in the body, such as for chronic use, and are considered medically insignificant for the particular investigational drug.

[0141] In one embodiment, two antigen-binding proteins are bioequivalent if there are no clinically significant differences in their safety, purity, and potency.

[0142] In one embodiment, two antigen-binding proteins are bioequivalent if a patient can be switched from a reference product to a biological product and vice versa one or more times without an expected increase in the risk of adverse effects, including a clinically significant change in immunogenicity or a decrease in efficacy, according to compared with continuous therapy without such translation.

[0143] In one embodiment, two antigen-binding proteins are bioequivalent if they both act through the same mechanism or mechanisms of action for the condition or conditions of use, to the extent such mechanisms are known.

[0144] Bioequivalence can be demonstrated using in vivo and in vitro methods. Options for evaluating bioequivalence include, for example, (a) an in vivo test in humans or other mammals that measures the evolution of the concentration of an antibody or its metabolites in blood, plasma, serum, or other body fluid; (b) an in vitro test that has been correlated with human in vivo bioavailability data and is reasonably predictive; (c) an in vivo test in humans or other mammals that measures the time course of the respective pharmacological action of a single dose of an antibody (or its target); and (d) in a well-controlled clinical trial that establishes the safety, efficacy, or bioavailability or bioequivalence of an antigen-binding protein.

[0145] Bioequivalent variants of the exemplary antibodies and bispecific antigen-binding molecules provided herein can be constructed, for example, by various residue or sequence substitutions, or by deletion of terminal or internal residues or sequences that are not necessary to exhibit biological activity. For example, cysteine residues that are not essential for biological activity can be deleted or replaced with other amino acids to prevent the formation of unnecessary or incorrect intramolecular disulfide bridges upon renaturation. In other contexts, bioequivalent antigen-binding proteins may include variants of the exemplary antibodies and bispecific antigen-binding molecules provided herein containing amino acid substitutions that modify the glycosylation characteristics of the molecules, such as mutations that eliminate or abolish glycosylation.

Therapeutic drug and delivery

[0146] The present invention relates to pharmaceutical compositions containing antigen-binding molecules according to the present invention. Pharmaceutical compositions according to the invention are formulated with suitable carriers, excipients and other agents that provide improved transfer, delivery, tolerability and the like. Many suitable drugs can be found in the reference book 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 (cationic or anionic) vesicles (such as LIPOFECTIN™, Life Technologies, Carlsbad, CA), DNA conjugates, anhydrous absorbent pastes, emulsions such as "oil in water" and "water in oil", emulsions based on carbovax (polyethylene glycols with different molecular weight), semi-solid gels and semi-solid mixtures containing carbovax. See also Powell et al. "Compendium of excipients for parenteral formulations" PDA (1998) J Pharm Sci Technol 52:238-311.

[0147] Various delivery systems are known and can be used to administer the pharmaceutical composition of the invention, for example, encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, receptor-mediated endocytosis (see, for example, Wu et al. , 1987, J Biol Chem 262:4429-4432). Routes of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The composition may be administered by any convenient route, for example by infusion or bolus injection, by absorption through epithelial or mucocutaneous integuments (e.g., oral mucosa, rectal and intestinal mucosa, etc.), and may be administered together with other biologically active agents. The introduction can be systemic or local. In one embodiment, the therapeutic proteins of the invention are administered by intravenous infusion.

[0148] The pharmaceutical composition of the present invention can be delivered subcutaneously or intravenously using a standard needle and syringe. In addition, in the case of subcutaneous delivery, a pen is very suitable for delivering the pharmaceutical composition of the present invention. Such a syringe pen can be reusable or disposable. A reusable pen typically uses a replaceable cartridge containing a pharmaceutical composition. Once all of the pharmaceutical composition has been introduced from the cartridge and emptied, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen can then be reused. There is no replaceable cartridge in the disposable syringe pen. In contrast, a disposable pen comes already filled with a pharmaceutical composition contained in a reservoir inside the device. Once the reservoir is empty of the pharmaceutical composition, the device is discarded as a whole.

[0149] A variety of reusable pens and auto-injectors are suitable for subcutaneous delivery of a pharmaceutical composition of the present invention. Examples include, but are not limited to, AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK), DISETRONIC™ pen (Disetronic Medical Systems, Burgdorf, Switzerland), HUMALOG MIX 75/25™ pen, HUMALOG pen ™, HUMALIN 70/30™ pen (Eli Lilly and Co., Indianapolis, IN), NOVOPEN™ I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR™ (Novo Nordisk, Copenhagen, Denmark), BD™ pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPEN™, OPTIPEN PRO™, OPTIPEN STARLET™ and OPTICLIK™ (sanofi-aventis, Frankfurt, Germany), and others. Examples of disposable pens suitable for subcutaneous delivery of a pharmaceutical composition of the present invention include, but are not limited to, SOLOSTAR™ pen (sanofi-aventis), FLEXPEN™ (Novo Nordisk) and KWIKPEN™ (Eli Lilly), SURECLICK ^™ autoinjector (Amgen, Thousand Oaks, CA), PENLET ^TM (Haselmeier, Stuttgart, Germany), EPIPEN (Dey, LP) and HUMIRA ^TM (Abbott Labs, Abbott Park, IL), and others.

[0150] In certain situations, the pharmaceutical composition may be delivered in 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 yet 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, the controlled release system can be placed in close proximity to the target of the composition, resulting in only a fraction of the systemic dose being required (see, for example, Goodson, 1984, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138). Other controlled release systems are reviewed by Langer, 1990, Science 249:1527-1533.

[0151] Injectable forms may include dosage forms for intravenous, subcutaneous, intradermal and intramuscular injections, drip infusions, etc. These injection forms can be obtained using well-known methods. For example, injectable forms may be prepared, for example, by dissolving, suspending or emulsifying an antibody or salt thereof, as described above, in a sterile aqueous or oily medium commonly used for injection. As an aqueous injection medium, there are, for example, saline, isotonic solution containing glucose and other auxiliary agents, etc., which can be used in combination with a suitable solubilizing agent such as alcohol (for example, ethanol), polyhydric alcohol (e.g., propylene glycol, polyethylene glycol), nonionic surfactant [e.g., polysorbate 80, HCO-50 (adduct of polyoxyethylene (50 mol) and hydrogenated castor oil)], etc. As the oil medium, for example, sesame oil, soybean oil, etc. are used, which can be used in combination with a solubilizing agent such as benzyl benzoate, benzyl alcohol, etc. The injection thus prepared is preferably filled into a suitable ampoule.

Therapeutic uses of antigen-binding molecules

[0152] The present invention includes methods comprising administering to a subject in need thereof a therapeutic composition comprising a therapeutic protein. Said therapeutic composition may contain any of the antibodies or bispecific antigen-binding molecules as disclosed herein and a pharmaceutically acceptable carrier or diluent. The term "subject in need thereof" means a human or non-human animal that exhibits one or more symptoms or signs of cancer (eg, a subject with evidence of a tumor or suffering from any of the cancers mentioned herein below).

[0153] The antibodies and bispecific antigen-binding molecules of the invention (and therapeutic compositions containing them) can be successfully used, inter alia, in the treatment of any disease or disorder that can benefit from stimulating, activating and/or targeting an immune response. In particular, the antibodies or bispecific antigen-binding molecules of the present invention may be used to treat, prevent and/or alleviate any disease or disorder associated with or mediated by the expression or activity of, for example, CD20, PSMA, MUC16, STEAP2 or BCMA, or the proliferation of CD20+, PSMA+, MUC16+, STEAP2+ or BCMA+ cells. The mechanisms of action on which the therapies of the invention are based include the killing of cells expressing such antigens in the presence of effector cells, for example, by CSC, apoptosis, ADCC, phagocytosis, or a combination of two or more of these mechanisms.

[0154] 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 meninges, head and neck, oropharynx, lung and bronchial tree, gastrointestinal tract, male and female reproductive organs pathways, muscles, bones, skin and appendages, connective tissue, spleen, immune system, hematopoietic cells and bone marrow, liver and urinary tract, kidney, bladder and/or special sense organs such as the eye. In certain embodiments, the antibodies and bispecific antigen-binding molecules of the invention are used to treat one or more of the following cancers, but are not limited to: pancreatic carcinoma, head and neck cancer, prostate cancer, malignant gliomas, osteosarcoma, colorectal cancer, gastric cancer (eg, MET-amplifying gastric cancer), malignant mesothelioma, multiple myeloma, ovarian cancer, small cell lung cancer, non-small cell lung cancer, synovial sarcoma, thyroid cancer, breast cancer, melanoma, glioma, breast cancer (eg, ductal or intraductal carcinoma of the breast), squamous cell carcinoma, esophageal cancer, clear cell renal cell carcinoma, chromophobe renal cell carcinoma, oncocytoma (kidney), transitional cell carcinoma (kidney), urothelial carcinoma, adenocarcinoma (bladder), or small cell carcinoma (urinary bubble). In certain embodiments of the present invention, antibodies or bispecific antibodies can be successfully used to treat a patient suffering from treatment-refractory or treatment-resistant cancer, such as castration-resistant prostate cancer.

[0155] In some embodiments, the antigen-binding molecule is a bispecific anti-CD3 x anti-CD20 antibody that can be successfully used to treat CD20-expressing cancer, 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's macroglobulinemia, primary mediastinal B-cell lymphoma, lymphoblastic lymphoma or Burkitt's lymphoma. In some embodiments, the cancer is follicular lymphoma. In some embodiments, the cancer is diffuse large B-cell lymphoma (DLBCL). In some embodiments, the cancer is mantle cell lymphoma. In some embodiments, the cancer is a marginal zone lymphoma. In some embodiments, the cancer is follicular lymphoma and the maximum weekly dose of the bispecific antibody is 80 mg. In some embodiments, the cancer is DLBCL and the maximum weekly dose of the bispecific antibody is 80 mg. In some embodiments, the cancer is DLBCL and the maximum weekly dose of the bispecific antibody is 160 mg. In some embodiments, the cancer is DLBCL and the maximum weekly dose of the bispecific antibody is 320 mg. In any of these or other embodiments discussed herein, the cancer patient may have a history of prior therapy with a monospecific anti-CD20 antibody.

[0156] Non-Hodgkin's lymphoma (NHL) is the most common hematological malignancy. Among the heterogeneous group of NHL, 85-90% are of B-cell origin and include follicular lymphoma (FL), diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), marginal zone lymphoma (MLZ), and several other B-NHLs. Anti-CD20 antibodies in combination with chemotherapy are the standard of care for B-NHL; however, despite an initial response, many patients relapse, often with progressively shorter response times on subsequent lines of therapy and poor outcomes. Thus, in some embodiments, the antigen binding molecule is an anti-CD3 x anti-CD20 bispecific that binds to CD3+ T cells and CD20+ B cells to target CD20+ tumor cells through T cell mediated cytotoxicity. In some cases, the bispecific anti-CD3 x CD20 antibody is for the treatment of B cell cancer (eg, NHL) in a subject who has failed prior monospecific anti-CD20 antibody therapy.

[0157] For patients with less than a complete response to CAR-T therapy, outcomes tend to be poor, and there are no standard therapy options. Thus, in some cases, the anti-CD3 x CD20 bispecific antibody of the present invention is for the treatment of B-cell cancer (e.g., NHL such as DLBCL) in a subject who has failed prior CAR-T therapy or is refractory to prior CAR. -T therapy (eg, anti-CD19 CAR-T therapy).

[0158] In some embodiments, the antigen binding molecule is a bispecific anti-CD3 x anti-PSMA antibody that can be successfully used to treat PSMA-expressing cancer, including prostate cancer, kidney cancer, bladder cancer, colorectal cancer, and gastric cancer. In some embodiments, the cancer is prostate cancer (eg, castration-resistant prostate cancer).

[0159] In some embodiments, the antigen-binding molecule is a bispecific anti-CD3 x anti-MUC16 antibody that can be successfully used to treat MUC16-expressing cancer, including ovarian cancer, breast cancer, pancreatic cancer, non-small cell lung cancer, intrahepatic cholangiocarcinoma in massive form, adenocarcinoma of the cervix and adenocarcinoma of the gastrointestinal tract. In some embodiments, the cancer is ovarian cancer.

[0160] In some embodiments, the antigen-binding molecule is a bispecific anti-CD3 x anti-STEAP2 antibody that can be successfully used to treat STEAP2-expressing cancer, including prostate cancer, bladder cancer, cervical cancer, lung cancer, colon cancer. colon, 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).

[0161] In some embodiments, the antigen binding molecule is a bispecific anti-CD3 x anti-BCMA antibody that can be successfully used to treat BCMA-expressing cancer, including multiple myeloma or other B-cell or plasma cell cancers, such as Waldenström 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's lymphoma. In some embodiments, the cancer is multiple myeloma.

Combination Therapies

[0162] The present invention provides methods of treatment comprising administering 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 can be used in combination with or administered in combination with an antigen binding molecule of the present invention include, for example, an antitumor agent (e.g., chemotherapeutic agents including melphalan, vincristine (Oncovin), cyclophosphamide (Cytoxan), etoposide (VP- 16), doxorubicin (Adriamycin), liposomal doxorubicin (Doxil), bendamustine (Treanda), or any other drugs found effective in the treatment of plasma cell tumors in the subject). In certain embodiments, the implementation of the second therapeutic agent is a regimen, including radiation therapy or transplantation of hematopoietic stem cells. In certain embodiments, the implementation of the second therapeutic agent may be an immunomodulator. 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 may be a monoclonal antibody, an antibody-drug conjugate, a bispecific antibody conjugated to an anticancer agent, an immune checkpoint inhibitor, or combinations thereof. Pharmaceutical compositions of the present invention (e.g., pharmaceutical compositions comprising a bispecific antigen-binding molecule as disclosed herein) may also be administered as part of a treatment regimen comprising one or more therapeutic combinations selected from a monoclonal antibody other than those described herein. , which can interact with another antigen on the surface of plasma cells, a bispecific antibody having one arm that binds to an antigen on the surface of tumor cells and another arm that binds to an antigen on a T cell, an antibody-drug conjugate, a bispecific antibody, an antineoplastic agent-conjugated, checkpoint inhibitor, eg targeted to PD-1 or CTLA-4, or combinations thereof. In certain embodiments, checkpoint inhibitors can be selected from PD-1 inhibitors such as pembrolizumab (Keytruda), nivolumab (Opdivo), or cemiplimab. In certain embodiments, checkpoint inhibitors can 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.

[0163] The present invention also includes therapeutic combinations containing any of the antigen-binding molecules mentioned herein and an inhibitor of one or more of VEGF, Ang2, DLL4, EGFR, ErbB2, ErbB3, ErbB4, EGFRvIII, cMet, IGF1R, B-raf , PDGFR-α, PDGFR-β, FOLH1 (PSMA), PRLR, STEAP1, STEAP2, TMPRSS2, MSLN, CA9, or uroplakin, wherein said inhibitor is an aptamer, antisense, ribozyme, siRNA, peptibody, nanobody, or antibody fragment (e.g. , Fab fragment; F(ab') ₂ fragment; Fd fragment; Fv fragment; scFv; dAb fragment; or other constructed molecules such as diabodies, triabodies, tetrabodies, minibodies, and minimal recognition units). Antigen-binding molecules according to the invention can also be introduced and/or included in the combination preparation with antivirals, antibiotics, analgesics and/or NSAIDs. The antigen-binding molecules of the invention may also be administered as part of a treatment regimen that also includes radiation therapy and/or conventional chemotherapy.

[0164] The additional therapeutically active component(s) may be administered immediately before, simultaneously with, or shortly after administration of the antigen-binding molecule of the present invention (in the context of the present invention, such administration regimens are considered to be the administration of the antigen-binding molecule “in combination with” the additional therapeutically active component) .

[0165] The present invention includes pharmaceutical compositions in which the antigen-binding molecule of the present invention is contained in combination with one or more additional therapeutically active ingredients as described elsewhere herein.

EXAMPLES

[0166] The following examples are provided 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 of this invention consider to be their invention. Efforts have been made to ensure accuracy with respect to the numbers used (eg amounts, temperatures, etc.), but the possibility of some experimental errors and deviations should be taken into account. Unless otherwise indicated, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric.

Example 1: Clinical evaluation and dose escalation of a bispecific antibody

[0167] The clinical trial described below is an open-label, multicentre, phase 1 study to investigate the safety and tolerability of REGN1979, a bispecific monoclonal anti-CD20 x anti-CD3 antibody, in patients with CD20+ B-cell malignancies previously treated with CD20-targeted antibodies.

[0168] Objectives : The main objectives of the study were to evaluate the safety, tolerability, and dose-limiting toxicity (DLT) of intravenous (IV) REGN1979 and to investigate the antitumor activity of REGN1979 in an expanded cohort including diffuse large B-cell lymphoma (DLBCL) after treatment failure Chimeric antigen receptor T cells (CAR-T), aggressive lymphoma (except DLBCL after failed CAR-T therapy), grade 1-3a follicular lymphoma (FL), and chronic lymphocytic leukemia (CLL). Additional objectives of the study were to: (1) characterize the pharmacokinetic (PK) profile of REGN1979; (2) 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 antibodies, or chronic lymphocytic leukemia [CLL]); and (4) to investigate the preliminary antitumor activity of REGN1979 in the dose escalation portion of the study. The assessment of minimal residual disease (MRD) was performed in patients with CLL. The exploratory goals of the study were to evaluate biomarkers that may correlate with the mechanism of action, observed toxicity, and potential antitumor activity, including but not limited to: (1) cytokine profiles and evaluation of inflammatory markers (eg, C-reactive protein [CRP]); (2) peripheral blood B cell and T cell subpopulations and immune phenotyping; and (3) changes in gene expression in peripheral blood.

[0169] Study Design : Patients were divided into dose level (DL) cohorts, which consisted of an initial starting dose and subsequent higher escalation doses for the second and subsequent dosing. Patients were included in the study based on disease (NHL or CLL). Each DL had 2 cohorts (one for each disease) comprising 3 to 6 patients per NHL cohort and 1 to 6 patients per CLL cohort. Patients with small lymphocytic lymphoma (MLL) were included in the CLL cohort and underwent the same evaluations as the NHL cohort.

[0170] Patients who initially showed clinical improvement and who subsequently relapsed, progressed, or showed a suboptimal response to treatment could be re-treated with REGN1979 at the highest DL considered tolerable at the time of relapse or progression.

[0171] Patients underwent screening procedures to determine eligibility for inclusion 28 days prior to initial administration of REGN1979. Patients were enrolled sequentially based on disease (NHL or CLL) in order of sponsor confirmation of eligibility until each cohort was completed according to protocol criteria.

[0172] Separate independent dose escalation cohorts were created for NHL and CLL at each DL. Each DL consisted of an initial dose and a second and subsequent doses that were higher than the starting dose, provided that the initial dose was tolerated.

[0173] Dose escalation was performed according to the traditional 3+3 dose escalation schedule for patients with NHL. Three to six patients were scheduled per cohort based on observed toxicity.

[0174] Dose escalation was performed according to a modified 3+3 schedule with an accelerated dose titration component for patients with CLL. Based on observed toxicity, 1 to 6 patients per cohort were planned.

[0175] At the end of the dose escalation phase and after determining the recommended dose for further study in patients with NHL, three expanded cohorts were to be opened for 1) patients with DLBCL after failed CAR-T therapy (20 patients), 2) aggressive lymphoma (except DLBCL after CAR-T failure) (40 patients; of which 20 patients were to be included in Cohort 1 [160 mg escalation]) and Cohort 2 aggressive lymphoma [80 mg escalation]), and 3) patients with relapsing /refractory follicular lymphoma grades 1-3a (60 patients; of which 30 patients each were to be included in cohort 1 [80 mg boost] and cohort 2 [160 mg boost]). The weekly escalation dose of REGN1979 for each of these expanded cohorts (except for aggressive lymphoma cohort 2 and follicular lymphoma cohort 1) was set at 160 mg per week, followed by maintenance treatment with 320 mg of REGN1979 qd/wk. In aggressive lymphoma cohort 2 and follicular lymphoma cohort 1, the weekly escalation dose of REGN1979 was 80 mg and the maintenance dose of REGN1979 1p/2wk was 160 mg. Patients assigned to an 80 mg escalation dose received 80 mg weekly for a 4-week induction period following an initial dose increase followed by an additional 8 weekly doses and treatment with 160 mg qd/2 wk until progression, with the option to discontinue treatment once the patient demonstrates sustained response within at least 9 months after initially achieving a complete response. Patients assigned to the 160 mg escalation dose received 160 mg weekly for a 4-week induction period following an initial dose increase followed by an additional 8 weekly doses and treatment with 320 mg qd/2 wk until progression, with the option to discontinue treatment once the patient demonstrates sustained response within at least 9 months after the initial demonstration of the software.

[0176] The feasibility of a pre-dose of rituximab prior to the first administration of REGN1979 was studied in patients with NHL to determine whether such an intervention could reduce the incidence and severity of infusion reaction (IR) and cytokine release syndrome (CRS). In the rituximab pre-dose cohort, REGN1979 was administered at an escalating dose of 160 mg during weekly treatment and 320 mg during 1r/2wk maintenance treatment. Then another 24 patients were to be treated using the optimal dosing and dose regimen; along with 6 patients in the rituximab pre-dose group treated with the optimal dose, a total of 30 patients were studied for safety and tolerability.

[0177] At the first DL, for the dose escalation portion between initial doses of study drug for the first 3 patients with the same disease, a mandatory 48-hour waiting period was observed. Subsequent patients on the first DL were not treated on the same day, regardless of disease. In subsequent cohorts, provided that no unexpected toxicity was observed in previous cohorts or in this cohort, the initial infusions for the first 3 patients were administered at least 24 hours apart.

[0178] After each cohort of patients was recruited, received treatment, and completed the observation period of ESWL, the decision was made to open subsequent DL cohorts to include patients (or expand the current open DL cohort) after the safety data were reviewed by the sponsor and the researcher(s).

[0179] The observation period of ESWL was defined as the first 28 days of treatment, which in this study corresponds to the induction period. During induction, patients received 4 weekly injections of REGN1979.

[0180] To be eligible for evaluation of ESWL, an individual patient must have received at least the first 2 injections of REGN1979 (day 1 of week 1 ["initial or primary dose"] and day 1 of week 2 ["secondary and subsequent dose"]), or try DLT. Patients enrolled on DL11 and above must have received at least the first 3 doses of REGN1979 (initial dose at week 1, intermediate or secondary dose at week 2, and a higher tertiary or escalation dose at week 1) or experience SWL. In addition, the patient had to be observed for at least 28 days after the first injection and at least 21 days after the second injection.

[0181] Dose escalation and NHL and CLL patient cohorts are shown in Table 1 below.

Table 1. Dose escalation and cohorts

Dose level Initial dose (mcg) Intermediate dose (mcg) Maximum dose (increasing dose) (mcg) NHL n HLL n DL1 thirty - 100 Cohort 1 3-6 Cohort 1 1-6 DL2 100 - 300 Cohort 2 3-6 Cohort 2 1-6 DL3 300 - 1000 Cohort 3 3-6 Cohort 3 1-6 DL4 1000 - 2000 Cohort 4 3-6 Cohort 4 1-6 DL5 1000 - 3000 Cohort 5 3-6 Cohort 5 1-6 DL6 1000 - 4000 Cohort 6 3-6 Cohort 6 1-6 DL7 1000 - 5000 Cohort 7 3-6 Cohort 7 1-6 DL8 1000 - 6000 Cohort 8 3-6 Cohort 8 1-6 DL9 1000 - 7000 Cohort 9 3-6 Cohort 9 1-6 DL10 1000 - 8000 Cohort 10 3-6 Cohort 10 1-6 DL11 1000 6000 12 000 Cohort 11 3-6 Cohort 11 1-6 DL12 1000 9000 18 000 Cohort 12 3-6 Cohort 12 1-6 DL13 1000 13 500 27 000 Cohort 13 3-6 Cohort 13 1-6 DL14 1000 20,000 40 000 Cohort 14 3-6 Cohort 14 1-6 DL15 1000 20,000 80 000 Cohort 15 3-6 Cohort 15 1-6 DL16 1000 20,000 160 000 Cohort 16 3-6 Cohort 16 1-6 DL17 1000 20,000 320 000 Cohort 17 3-6 Cohort 17 1-6

[0182] In this study, each dose level includes an initial dose of REGN1979 followed by an increasing dose; for cohorts DL11 and above, a secondary dose was added until a booster dose was reached (see Table 1). Each of the initial (primary) dose, the intermediate (secondary) dose (if applicable), and the first dose of the boosting (tertiary) dose may optionally be administered in multiple doses over at least 2 days. All patients in the maximum dose cohorts of DL7 and above received split dosing (eg, dosing in installments) for the initial (primary) and intermediate (secondary) doses. Patients in the maximum dose cohorts of DL11 and above received split dosing (eg, dosing in installments) for the initial (primary), intermediate (secondary), and first boost (tertiary) doses followed by booster doses (maximum weekly dose).

[0183] Duration of the study : the treatment period is 9 months. Patients will receive 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 twice-monthly (1p/2wk) doses for a maintenance period until progression, with an option to discontinue treatment after 9 months after the patient achieves a complete response. Patients may be followed up for efficacy and safety for up to 15 months after the end of treatment.

[0184] Study Population : Assuming study enrollment up to DL17 and full enrollment in all expanded cohorts, up to 370 patients are planned at approximately 15 centers in the US and Germany. Up to 204 patients will be included in dose escalation cohorts up to DL17 for both diseases (NHL and CLL) during the dose escalation phase. Up to 100 patients, including 90 patients with NHL (20 patients with DLBCL after failed CAR-T therapy, 40 patients with aggressive lymphoma [excluding DLBCL after failed CAR-T therapy], 30 patients with grade 1-3a FL) and 10 patients with CLL will be included in expanded disease cohorts. Up to 42 patients will be included in the rituximab prep dose cohort to determine the optimal dosing regimen. Once the optimal rituximab pre-dose schedule has been established, an additional 24 patients will be included in the expanded rituximab pre-dose cohort and will be pooled with 6 patients from the above rituximab pre-dose cohort who received the optimal dosing schedule and dose. Thus, a total of 30 patients will be studied at the optimal rituximab prep dose schedule and dose.

[0185] Patients must have a confirmed CD20+ B cell malignancy with active disease, refractory to prior therapy, for which there are no standard treatment options, and for which anti-CD20 antibody treatment may be appropriate. Patients with NHL should have a history of anti-CD20 antibody therapy.

[0186] Inclusion Criteria : A patient must meet the following criteria to be eligible to participate in the study:

1. Have a documented CD20+ B-cell malignancy with active disease, refractory to prior therapy, for which there are no standard treatment options, and for which anti-CD20 antibody treatment may be appropriate:

B-NHL validated by NCI Working Group criteria (National Cancer Institute), 2007 (Cheson 2007, Appendix 2); And

CLL validated by the 2008 International Workshop on Chronic Lymphocytic Leukemia (IWCLL) working group criteria (Hallek 2008, Annex 3) - patients with small cell lymphocytic lymphoma (SLL) will be included in the CLL cohort and will undergo the same assessments as the NHL cohort.

Note: A patient with a CD20-negative lymph node biopsy (NHL) performed as standard of care just prior to study entry is still eligible for study provided they have previously confirmed CD20+ disease AND HAVE previously received treatment with rituximab or other anti-CD20 antibody therapy for about 6 months. Individual cases may be discussed with the medical supervisor.

2. Patients with NHL should have a history of anti-CD20 antibody therapy. Patients with CLL are not required to have a history of anti-CD20 antibody therapy provided that the patient has failed therapy with a BTK inhibitor or a PI3K inhibitor and the treating physician considers it appropriate to enroll the patient in a phase 1 study. To be included in the expanded Grade 1-3a FL cohort, patients must have received at least 2 previous lines of systemic therapy, including anti-CD20 antibody and an alkylating agent. To be included in an extended disease cohort of patients with DLBCL after failed CAR-T therapy, the patient must recover from the toxic effects of lymphodepletion therapy and CAR-T infusion. Prior CAR-T therapy does not have to be the last line of therapy used prior to enrollment in the study.

3. All patients (with B-cell NHL and CLL) must have at least one bidirectionally measurable lesion ≥1.5 cm) confirmed by CT scan or MRI if CT scan is not available.

4. CLL patients must have a white blood cell count (WBC) ≤200×109/L

5. Age ≥18 years old

6. Assessment of the functional state according to the scale of the Eastern United Group of Oncologists (ECOG) ≤1

7. Life expectancy of at least 6 months

8. Acceptable bone marrow function, confirmed by: a. Platelet count ≥75×109/l; b. Hemoglobin level ≥9 g/dl; c. Absolute neutrophil count (ACN) ≥1×109/l

Note: Patients with cell counts below the cut-offs above may be considered for inclusion in the study if, in the opinion of the investigator, the cause is bone marrow infiltration from an underlying disease. In such cases, the investigator should discuss eligibility with the sponsor and obtain written permission to enter the study.

9. Acceptable organ function confirmed by:

• Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels ≤2.5 X ULN

• Total bilirubin ≤1.5 X ULN

Note: Patients with Gilbert's syndrome may not meet this requirement provided that their total bilirubin has not changed from baseline.

• Estimated creatinine clearance according to the Cockcroft-Gault formula ≥50 ml/min

Note: Patients may be considered for inclusion in the study if, in the opinion of the investigator, abnormal laboratory results are related to the underlying disease. In such cases, the investigator should discuss eligibility with the sponsor and obtain written permission to enter the study.

Note: Patients with borderline Cockcroft-Gault creatinine clearance 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 prior tumor biopsy if, in the investigator's opinion, the patient has an accessible lesion that can be biopsied without significant risk to the patient.

11. Desire and readiness to follow the schedule of clinic visits and undergo procedures related to the study

12. Providing signed informed consent.

[0187] Exclusion Criteria : A patient meeting any of the following criteria will be excluded from the study:

1. Primary central nervous system (CNS) lymphoma or known or suspected CNS involvement of non-primary CNS NHL

2. Previous or current relevant CNS pathology such as

• Epilepsy, seizures, paresis, aphasia, apoplexy, severe brain injury, cerebellar disease, organic brain syndrome, psychosis, or

• Signs of inflammatory lesions and/or vasculitis on brain MRI

3. Standard antineoplastic chemotherapy (non-biologic) for 5x half-life or 28 days, whichever is shorter, prior to first administration of study drug.

4. Standard radiotherapy within 14 days prior to first administration of study drug.

Note: Palliative radiotherapy of a symptomatic lymph node/lesion is permitted provided that the irradiated lesion(s) or node(s) is not accepted as the target lesion for tumor assessment

5. Allogeneic stem cell transplant

6. Treatment with rituximab, alemtuzumab, or other experimental or commercial biological agent within 12 weeks prior to first administration of study drug.

Note: For patients with aggressive lymphoma requiring immediate treatment, the washout period may be reduced to 28 days. This will require discussion with the sponsor and written approval by the sponsor.

7. Immunosuppressive therapy (other than biological) within 28 days prior to first administration of study drug.

8. Treatment with an experimental non-biological agent within 28 days prior to the first administration of study drug.

9. History of allergic reactions associated with compounds with a chemical or biological composition similar to the study drug.

10. History of hypersensitivity to any compound from the group of tetracycline antibiotics.

11. Concomitant active malignancy for which the patient is being treated.

12. Identified active bacterial, viral, fungal, mycobacterial or other infection, or any serious episode of infection requiring hospitalization or treatment with intravenous anti-infective drugs within 4 weeks prior to the first injection.

13. Evidence of a significant comorbid disease or condition that could interfere with the study or put the patient at significant risk, including, but not limited to, serious cardiovascular disease (e.g., class III or IV cardiovascular disease according to the New York Heart Association, myocardial infarction in the previous 6 months, unstable arrhythmias or unstable angina) and/or serious lung disease (eg, obstructive pulmonary disease and a history of symptomatic bronchospasm).

Note: Patients with a history of cardiac disease should be evaluated by echocardiography or radionuclide ventriculography (MUGA) prior to the first administration of REGN1979 to ensure acceptable cardiac reserves and function.

14. Current treatment with systemic corticosteroids, except for the use of corticosteroids for other indications (not related to tumors and immunosuppression), up to a maximum dose of 10 mg / day of prednisone or its equivalent.

15. Infection with human immunodeficiency virus (HIV) or chronic infection with hepatitis B virus (HBV) or hepatitis C virus (HCV). Hepatitis B (HepBsAg+) patients who have a controlled infection (serum HBV DNA level below the detection limit AND ALSO receiving hepatitis B antiviral therapy) are eligible in consultation with the treating physician treating the infection.

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 before starting study drug treatment, during the study, and for at least 6 months after the last dose. Highly effective contraceptive measures include regular use of combined (estrogen and progestogen) hormonal contraception (oral, intravaginal, transdermal) or progestogen-only hormonal contraception (oral, injectable, implantable) associated with inhibition of ovulation starting 2 or more menstrual cycles prior to screening; intrauterine device; intrauterine hormonal system; bilateral tubal ligation; partner vasectomy; and/or sexual abstinence†, ‡.

*Postmenopausal women must have been amenorrheic for at least 12 months to be considered infertile. Pregnancy testing and contraception are not required for women with a confirmed hysterectomy or tubal ligation.

† Sexual abstinence is considered highly effective only if it is defined as abstaining from heterosexual intercourse during the entire period of risk associated with investigational treatment. The reliability of sexual abstinence needs to be assessed in relation to the duration of the clinical study and the patient's preferred and usual lifestyle.

‡ Intermittent abstinence (calendar, symptomatic, post-ovulation methods), coitus withdrawal (coitus interruptus), spermicide use alone, and the lactational amenorrhea method (LAM) are not acceptable methods of contraception. The female condom and the male condom should not be used together.

19. Administering a live vaccine within 28 days prior to the first administration of study drug

20. A member of the clinical trial site research team and/or his/her next of kin, unless prior authorization has been given by the sponsor.

[0188] Treatment : REGN1979 was supplied as a liquid in sterile disposable vials. Each vial contained REGN1979 at a concentration of 2 mg/mL. Detailed instructions for preparation and use have been provided to the clinical study sites in the Pharmaceutical Manual. A diluent was supplied for the preparation of study drug REGN1979.

[0189] Patients received REGN1979 weekly for a 4-week induction period followed by 8 weekly doses and 1p/2wk doses until progression at a dose appropriate to their assigned cohort.

[0190] Only the rituximab prep dose cohort and the extension cohort received a single dose of rituximab (375 mg/m ² ) one day prior to the first dose of REGN1979 [i.e. on (-1) study day]. REGN1979 was started on day 1 of week 1 and the REGN1979 treatment period was 9 months. Patients received up to 24 doses of REGN1979: 4 weekly doses during a 4-week induction period followed by an additional 8 weekly doses and 12 or more doses given 1p/2wk during a maintenance period until progression. In the rituximab pre-dose cohort, REGN1979 was administered at an escalating dose of 160 mg during weekly treatment and 320 mg during 1r/2wk maintenance treatment. Dose groups with increasing doses of REGN1979 below 160 mg for weekly treatment and below 320 mg for REGN1979 maintenance treatment 1r/2wk can also be evaluated. An additional 24 patients were then evaluated at the optimal dosing schedule and dose; along with 6 patients in the rituximab prep dose group who received the optimal dosing regimen and dose, a total of 30 patients were studied for safety and tolerability.

[0191] Endpoints

[0192] Primary : The primary endpoints were safety (particularly adverse events [AEs] and EBRT) to determine the maximum tolerated dose (MTD) and/or optimal biological dose (OBD) as the recommended dose of REGN1979 for phase 2 (RP2D) ; and efficacy as measured by objective response rate (ORR) in an expanded cohort of patients with DLBCL after failed CAR-T therapy, in expanded cohorts 1 and 2 of aggressive lymphoma (except DLBCL after failed CAR-T therapy), an expanded cohort of FL grades 1-3a and the extended CLL cohort.

[0193] Secondary : Secondary endpoints were as follows:

• Pharmacokinetics: REGN1979 concentration

• Immunogenicity: antibodies to REGN1979

• Antitumor activity:

− Objective response rate (ORR)

• Evaluation of tumor response according to the Revised Criteria for Evaluation of Response in Malignant Lymphomas of the NCI International Working Group (NCI-WG)

• Evaluation of tumor response in accordance with the Guidelines for the Diagnosis and Treatment of CLL of the International Seminar on Chronic Lymphocytic Leukemia

• For patients included in extended NHL cohorts, assessment of tumor response according to the Lugano classification

− Progression-free survival (PFS) and overall survival (OS)

− Minimal Residual Disease (MRD) for CLL patients

[0194] Search endpoints include :

• Pharmacodynamic (PD) indicators, including:

− Subpopulations and phenotype of B-cells and T-cells

− Levels of circulating cytokines in the blood

− SRB

− Changes in gene expression in peripheral blood

[0195] Procedures and assessments

[0196] Baseline procedures : Brain MRI, electrocardiogram (ECG), testing for human immunodeficiency virus (HIV), hepatitis C virus (HCV), and hepatitis B virus (HBV), and coagulogram.

Safety procedures: history, physical examination, symptom assessment, assessment of general condition, clinical laboratory tests, vital signs, AEs, and concomitant medications.

[0197] Efficacy Evaluation Procedures : Tumor evaluation, including computed tomography or MRI, 18F-fluorodeoxyglucose (FDG-PET) positron emission tomography scan, bone marrow aspirate and biopsy, lymph node and/or tumor biopsy, and specimens peripheral blood (only for patients with CLL).

Blood samples were taken for evaluation of FC and anti-drug antibodies (ADA).

Biomarker samples were taken to monitor changes in cytokine production, serum levels of pro-inflammatory cytokines, and changes in subpopulations and lymphocyte activation status. In addition, these samples allowed for tumor or somatic genetic analysis of variations that influence the clinical course of the underlying disease or modulate the side effects of treatment.

[0198] Statistical Analysis Design : The study design was based on the traditional 3+3 design with 3-6 patients per DL for patients with NHL, and a modified 3+3 design with an accelerated dose titration component with 1-6 patients per DL for patients with HLL. The exact number of patients included in the study will depend on the number of patients (with NHL and CLL) observed with a specific EBRT protocol, and on treatment-related toxicity of Grade 2 or higher, at which acute effects (excluding associated laboratory abnormalities) resolve to ≤ grade 1 or baseline within 72 hours (CLL during the accelerated dose titration component), and the need to expand the currently defined DLs or open additional cohorts with lower DLs.

[0199] Patient recruitment is ongoing and up to 370 patients are planned. Up to 204 patients will be included in dose escalation cohorts up to DL17 for both diseases (NHL and CLL) during the dose escalation phase. Up to 100 patients, including 90 patients with NHL (20 patients with DLBCL after failed CAR-T therapy, 40 patients with aggressive lymphoma [excluding DLBCL after failed CAR-T therapy] and 30 patients with grade 1-3a FL) and 10 patients with CLL will be included in the expanded disease cohorts. Up to 42 patients will be included in the rituximab prep dose cohort to determine the optimal dosing regimen. An additional 24 patients will be included in the expanded rituximab prep dose cohort and will be pooled with 6 patients from the above rituximab prep dose cohort who received the optimal dosing regimen and dose. Thus, a total of 30 patients will be studied at the optimal rituximab prep dose schedule and dose.

[0200] The data are summarized using only descriptive statistics. Typically, data are aggregated by DL and by disease (NHL or CLL). Within NHL disease, data will also be summarized by subgroup and dose for DLBCL after CAR-T failure, cohort 1 and 2 aggressive lymphoma (except DLBCL after CAR-T failure), and grade 1-3a FL. Within NHL disease, data are also summarized by subgroups of indolent and aggressive NHL. Patient population characteristics and baseline characteristics are summarized across groups using descriptive statistics.

[0201] Generalizations and safety analyzes were performed on a sample for safety analysis (SAF). The primary safety analysis was based on treatment-emergent AEs (AEs). Based on this analysis, conclusions were drawn regarding the safety profile of REGN1979. All AEs reported in this study were coded using the current available version of the Medical Dictionary for Regulatory Practice (MedDRA®). Encoding will be done in terms of the lower level. Coded text, preferred term (PT), and major organ system class (SOC) were specified. Analysis of efficacy and baseline variables is performed using a performance analysis sample (FAS). Efficacy analyzes for expanded cohorts including DLBCL after CAR-T failure, aggressive lymphoma cohorts 1 and 2 (except DLBCL after CAR-T failure), FL grades 1-3a, and CLL will be performed separately after all patients in eligible cohort will visit at week 24 or withdraw from the study before that time.

[0202] Results : The results for patients who received the maximum weekly dose of 5 to 320 mg indicate a low incidence of CRS. Serum concentrations of REGN1979 in patients treated with a maximum weekly dose of only 12 mg were found to approach or exceed serum levels demonstrated to be effective in Raji tumor xenograft mouse models (data not shown). Patients receiving a maximum weekly dose of as little as 40 mg maintained a serum concentration above the trough concentration (2000 µg/L) shown to be effective in mouse xenograft Raji tumor models (data not shown). In addition, patients treated at these levels also experienced multiple partial and complete responses, as shown in Tables 2-9 below. The pooled safety and PK data for REGN1979 during the 13N cohort EBRT evaluation period (27,000 µg REGN1979) demonstrate that the CRS or IR control algorithm (i.e., stepwise dose escalation, split dosing during the first weeks of REGN1979 administration, and premedication with corticosteroids) has proven its worth. Efficacy in preventing severe CRS or IR despite increasing dose in each successive dose cohort. Split dosing provided improved patient safety at weeks 1-4 (data available), where there was a lower cumulative incidence of severe CRS/IR. In particular, the dosing strategy discussed herein provided a safer strategy for increasing doses to levels greater than 80 mg, even 160 mg or more, with fewer severe events at weeks 3 and 4 when higher doses achieved and exceeded desired serum concentrations. discussed above. In FIG. 1 shows the incidence of CRS/IR in patients receiving the maximum dose of 320 mg. To date, no patients have withdrawn from the study due to an adverse event of CRS/IR.

Table 2 Observed response in grade 1-3a follicular lymphoma

CD20xCD3 ^† <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) Stable disease, 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 achieved) 11.8 (4.37-11.83) N/A

^† No patient received an 80 mg dose of REGN1979

[0203] After cessation of data collection (Table 2), two more evaluable patients showed complete responses (CR), one at 40 mg and the other at 320 mg.

Table 3 Observed response in diffuse large B-cell lymphoma

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) Overall response rate, 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/impossible to assess, 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 achieved) N/A N/A N/A

[0204] Two of the three patients treated with 80 mg who demonstrated PO had ineffective CAR T cell therapy. All complete responses indicated in Table 3 were complete metabolic responses.

[0205] All POs at doses of 80 mg, 160 mg, and 320 mg are current POs when receiving study treatment, indicating sustained response.

Table 4 Observed response in DLBCL after failed CAR-T therapy

CD20xCD3 3 mg
(N=1) 27 mg
(N=1) 40 mg
(N=1) 80 mg
(N=3) 160 mg
(N=1) Overall response rate, n (%) 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/impossible to assess, n (%) 0 0 0

Table 5 Observed response in mantle cell lymphoma

CD20xCD3 5-12 mg
(N=1) 18-40 mg
(N=1) 160 mg
(N=1) Objective response (PO/OR) 1 (100%) 1 (100%) 0 Full answer 0 1 (100%) 0 Partial answer 1 (100%) 0 0 stable disease 0 0 0 progressive disease 0 0 0 Missing/impossible to evaluate 0 0 1 (100%)

Table 6 Observed response in marginal zone lymphoma

CD20xCD3 5-12 mg
(N=1) 18-40 mg
(N=1) 80 mg
(N=3) Objective Answer
(PO/CH) 0 1 (100%) 2 (66.7%) Full answer 0 0 2 (66.7%) Partial answer 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 (PO/OR) 0 1 (100%) 0 1 (100%) 1 (100%) 1 (100%) Full answer 0 0 0 0 1 (100%) 1 (100%) Partial answer 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/impossible to rate 0 0 0 0 0 0

Table 8. Responses of LMZ Patients by Dose Level

CD20xCD3 4 mg
(N=1) 5 mg
(N=1) 27 mg
(N=1) 80 mg
(N=3) Objective response (PO/OR) 1 (100%) 0 1 (100%) 2 (66.7%) Full answer 0 0 0 2 (66.7%) Partial answer 1 (100%) 0 1 (100%) 0 stable disease 0 0 0 0 progressive disease 0 1 (100%) 0 1 (33.3%) Missing/impossible to rate 0 0 0 0

Table 9. Responses in patients with other NHL according to dose level

CD20xCD3 4 mg
(N=1) 12 mg
(N=1) 18 mg
(N=1) 27 mg
(N=1) NHL subtype FL of unknown degree macroglobulinemia
waldenstrom FL degree 3b FL of unknown degree Objective response (PO/OR) 0 0 1 (100%) 0 Full answer 0 0 1 (100%) 0 Partial answer 0 0 0 0 stable disease 1 (100%) 0 0 0 progressive disease 0 0 0 0 Absent/
impossible to assess 0 1 (100%) 0 1 (100%)

[0206] In general, broad spectrum activity observed in heavily pretreated patients with relapsed/refractory B-NHL treated with REGN1979, including some patients with progression from prior CAR T cell therapy, included:

(i) FL grades 1-3a: 12/13 (92.3%) ORR; 8/13 PO (61.5%) at doses ≥5 mg;

(ii) DLBCL: 4/6 (66.7%) ORR (all CR) at doses of 80-160 mg, with 2 patients achieving CR after failing CD19-targeted CAR T cell therapy;

(iii) SCL: 3/3 responses at doses ≥5 mg, including one CR;

(iv) LMZ: 3/5 responses at doses ≥5 mg, including two CRs;

(v) Tolerability in patients with B-NHL has been demonstrated up to doses of 320 mg per week, without any EBRT in patients with B-NHL;

(vi) Most adverse events were 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 patient discontinued treatment due to CRS or neurological adverse events. Of the 96 patients at baseline, only seven developed grade 3 CRS; And

(ix) Dexamethasone did not suppress the cytotoxicity of REGN1979, had little effect on the upregulation of T cell activation, and inhibited the release of cytokines.

Example 2: Clinical Evaluation of a Bispecific Antibody

[0207] The clinical study described below is an open-label, multicentre, phase 2 study to evaluate the antitumor activity and safety of REGN1979, a bispecific anti-CD20 x anti-CD3 antibody, in patients with relapsed or refractory follicular lymphoma.

[0208] Objectives : The main objective of this study was to evaluate the antitumor activity of REGN1979 as a monotherapy based on the objective response rate (ORR) according to the Lugano classification of response in malignant lymphoma (Cheson, 2014) using an independent centralized assessment in patients with follicular lymphoma ( FL) relapsed after at least two previous lines of systemic therapy, including an anti-CD20 antibody and an alkylating agent, or refractory to them. Additional objectives in this study were: (1) to evaluate the antitumor activity of REGN1979 as monotherapy in patients with relapsed or refractory FL based on (a) ORR according to the Lugano classification (Cheson, 2014) as assessed by a local investigator, (b) complete response rates (CR) according to the Lugano classification, by an independent central assessment and assessment by a local investigator, (c) progression-free survival (PFS) according to the Lugano classification, by an independent central assessment and assessment by a local investigator, (d) overall survival (OS), (e) Duration of Response (DR) according to Lugano classification, as assessed by an independent central assessment and assessment by a local investigator, (f) frequency of disease control (FCC) according to Lugano classification, by an independent central assessment and assessment by a local investigator , (g) duration of disease control (DDC) according to the Lugano classification, as assessed by an independent centralized assessment and assessment by a local investigator; (2) evaluate the safety and tolerability of REGN1979; (3) evaluate the pharmacokinetics (PK) of REGN1979; (4) assess the immunogenicity of REGN1979; and (5) assess the impact of REGN1979 on quality of life as measured by approved tools: the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30) and the European 5 Measures and 3 Levels Quality of Life Questionnaire (EQ). -5D-3L).

[0209] Study design : The study consists of a screening period of up to 28 days, a total treatment period of up to 98 weeks, which includes 12 weekly (1r/wk) doses, followed by once every 2 weeks (1r/2wk) dosing at up to 86 weeks, and a follow-up period of 96 weeks.

[0210] 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 then a nominal (tertiary and maximum weekly) dose of 80 mg, according to the scheme dosing of 12 injections 1r/week followed by dosing of 80 mg REGN1979 1r/2week.

[0211] Recruitment is conducted in accordance with the design of an open non-comparative study.

[0212] The screening period begins with the signing of the Informed Consent Form (ICF) and ends when the patient is confirmed to be eligible for inclusion in the study and begins treatment, or when it is determined that the patient is ineligible and was found not to have passed the screening.

[0213] The treatment period begins with the initial administration of REGN1979 and consists of 12 infusions of REGN1979 1r/wk followed by a dose of 2r/wk for 86 weeks, so that the total treatment period is 98 weeks of study drug administration, unless the patient stops treatment with study drug drug due to disease progression, initiation of subsequent lymphoma therapy, an adverse event (AE), or any other cause.

[0214] The follow-up period will be 96 weeks after the last dose of study drug. All patients will be monitored every 12 weeks to determine survival to death, loss of communication with the patient, patient withdrawal of consent to follow-up, or termination of the study by the sponsor, whichever comes first. For patients who discontinue study drug for any reason other than disease progression, initiation of subsequent lymphoma therapy, or death, disease response will be assessed every 12 weeks during the follow-up period until disease progression, death, initiation of subsequent lymphoma therapy, or patient withdrawal of consent for follow-up, whichever comes first.

[0215] Study Duration : The study duration for each patient, excluding the screening period, will be approximately 194 weeks, unless the patient demonstrates disease progression or initiation of subsequent therapy, or until death occurs, loss of communication with the patient, patient withdrawal of consent for follow-up observation or termination of the study by the sponsor. The end of the study is the last visit of the last patient.

[0216] Study Population : Up to 481 patients at up to 100 centers will be included in the study. The study population will consist of patients 18 years of age or older who have previously received treatment with grade 1 to 3a FL that has relapsed with or is refractory to at least two previous lines of systemic therapy, including an anti-CD20 antibody and an alkylating agent. . Prior to enrollment in the study, histopathological confirmation of the diagnosis of FL by central evaluation will be required. Patients with grade 3b FL are not eligible for inclusion in the study. Refractory disease is defined as no response to standard treatment or progression within 6 months of last treatment.

[0217] Inclusion Criteria : Each patient must meet the following criteria to be eligible for inclusion in the study:

1. Age 18 and over

2. Before enrollment in the study, histopathological confirmation of the diagnosis of Grade 1 to 3a FL should be obtained by centralized assessment. Patients with grade 3b FL are not eligible for inclusion in the study. The subtype definition of follicular lymphoma is based on the World Health Organization (WHO) classification (Swerdlow, 2017).

3. The disease must have relapsed or must have been refractory to ≥2 previous lines of systemic therapy, including anti-CD20 antibody and an alkylating agent. In the opinion of the investigator, patients should be in need of FL therapy at the time of enrollment in the study.

4. Measurable manifestations of disease on cross-sectional imaging (defined as at least 1 bidirectionally measurable nodular lesion ≥1.5 cm in greatest transverse section (GTD), regardless of short section size) confirmed by diagnostic imaging ( computed tomography [CT] or magnetic resonance imaging [MRI]).

5. Assessment of the functional state according to the scale of the Eastern United Group of Oncologists, equal to 0 or 1.

6. Acceptable bone marrow function confirmed by: (a) platelet count ≥50×10 ⁹ /L. To meet the platelet eligibility criteria, the patient must not have received a platelet transfusion within 7 days prior to the first dose of REGN1979; (b) hemoglobin ≥9.0 g/dl; (c) absolute neutrophil count (ANC) ≥1.0×10 ⁹ /l. To meet the ANC eligibility criteria, the patient must not have received granulocyte colony stimulating factor within 2 days prior to the first dose of REGN1979.

7. Acceptable liver function: (a) total bilirubin ≤1.5 × upper limit of normal (ULN) (≤3 × ULN if associated with liver lymphoid infiltration); (b) alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels ≤2.5×ULN (≤5×ULN if associated with liver lymphoid infiltration); (c) alkaline phosphatase (AP) level ≤2.5 × ULN (≤5 × ULN if associated with liver lymphoid infiltration); NOTES: Regardless of the presence of lymphoid infiltration of lymphoma, a patient with AST >2.5 x ULN and/or ALT >2.5 x ULN concomitantly with total bilirubin >1.5 x ULN will be excluded, and patients with confirmed Gilbert's syndrome may not qualify. requirement for total bilirubin, provided that the value has not changed from baseline.

8. Serum creatinine ≤1.5 × ULN, or estimated Cockcroft-Gault creatinine clearance ≥50 ml/min; NOTE: Patients with an estimated creatinine clearance <50 ml/min may be considered for inclusion in the study if the measured creatinine clearance (based on 24-hour urine or other reliable method) is ≥50 ml/min.

9. Willingness to undergo a tumor biopsy at baseline. If the investigator determines that a baseline tumor biopsy cannot be performed safely, the sponsor may make an exception to the biopsy requirement only after discussion and approval by the medical supervisor.

10. Ability to understand the purpose and risks of the study and provide signed and dated informed consent and permission to use protected health information (in accordance with national and local privacy laws).

11. Desire and readiness to follow the schedule of visits to the clinic and undergo procedures related to the study.

12. Providing informed consent signed by the patient or legal representative.

13. Ability to understand and complete questionnaires related to the study.

[0218] 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 CNS involvement of non-primary CNS NHL (if CNS lymphoma is suspected, routine lumbar puncture analysis should be done in addition to mandatory head CT or MRI).

2. Treatment with any systemic antilymphoma therapy for 5 half-lives or within 28 days prior to first administration of study drug, whichever is shorter.

3. History of allogeneic stem cell transplantation.

4. Prior treatment with any chimeric antigen receptor (CAR-T) T cell therapy.

5. Continuous treatment with systemic corticosteroids with administration of more than 10 mg of prednisone or its anti-inflammatory equivalent per day within 72 hours prior to the start of study drug.

6. History of neurodegenerative disease or CNS motor dysfunction. History of uncontrolled seizure disorder, defined as any seizure within 12 months prior to study entry.

7. Within 28 days prior to the first administration of the study drug, vaccination with a vector with replication potential.

8. Other malignancy other than FL in the past 5 years, excluding non-melanoma skin cancer that has undergone potentially curative therapy, or cervical carcinoma in situ, or any other tumor that has been found to be effectively treated by radical local control and with a curative intent.

9. Evidence of a significant comorbid disease or condition that could interfere with the study or put the patient at significant risk, including, but not limited to, serious cardiovascular disease (e.g., New York Heart Association class III or IV cardiovascular disease, myocardial infarction in the previous 6 months, unstable arrhythmias or unstable angina) and/or serious lung disease (eg, obstructive pulmonary disease and a history of symptomatic bronchospasm).

10. Ejection fraction of the heart <40% according to echocardiography or radionuclide ventriculography (MUGA).

11. Any infection requiring hospitalization or treatment with intravenous anti-infective drugs within 2 weeks prior to first administration of study drug.

12. Uncontrolled infection with human immunodeficiency virus (HIV), hepatitis B or hepatitis C; or other uncontrolled infection, except for the following: (a) HIV patients who have a controlled infection (undetectable viral load and CD4 count above 350 cells/microliter, either spontaneously or on an ongoing antiviral regimen) are eligible; (b) patients with hepatitis B (HepBsAg+) who have a controlled infection (serum hepatitis B virus DNA level below the polymerase chain reaction [PCR] detection limit AND ALSO receiving hepatitis B antiviral therapy) are eligible; (c) patients with antibodies to hepatitis C virus (HCV Ab+) who have a controlled infection (HCV RNA not detectable by PCR, either spontaneously or in response to a successful previous course of anti-HCV therapy) are eligible.

13. History of severe allergic reaction associated with compounds with a chemical or biological composition similar to the study drug or excipient. In this context, a severe allergic reaction is defined as a reaction requiring hospitalization and/or treatment with epinephrine.

14. Known hypersensitivity to both allopurinol and rasburicase.

15. A member of the clinical trial site research team or his/her next of kin, unless prior authorization has been given by the sponsor.

16. Women with a positive pregnancy test result for serum β-hCG at the screening visit. If the result is positive, pregnancy must be ruled out by ultrasound in order for the patient to participate in the study.

17. Patients placed in a medical institution on the basis of a decision of the judicial or administrative authorities.

18. Pregnant or lactating women.

19. Women of childbearing potential* or men who do not wish to use highly effective contraception prior to the initial dose/start of the first treatment, during the study, and for at least 6 months after the last dose. Highly effective contraceptive measures include: (a) regular use of combined (estrogen and progestogen) hormonal contraception (oral, intravaginal, transdermal) or progestogen-only hormonal contraception (oral, injectable, implantable) associated with inhibition of ovulation commencing 2 or more menstrual periods. cycles before screening; (b) an intrauterine device (IUD); intrauterine hormone-releasing system (HCV); (c) bilateral tubal ligation; (d) partner's vasectomy; (e) and/or sexual abstinence ^† , ^‡ .

*Postmenopausal women must have been amenorrheic for at least 12 months to be considered infertile. Pregnancy testing and contraception are not required for women with a confirmed hysterectomy or tubal ligation.

^† Sexual abstinence is considered highly effective only if it is defined as abstaining from heterosexual intercourse during the entire period of risk associated with investigational drugs. The reliability of sexual abstinence must be assessed in relation to the duration of the clinical study and the subject's preferred and usual lifestyle.

^‡ Intermittent abstinence (calendar, symptomatic, post-ovulation methods), coitus withdrawal (coitus interruptus), spermicide use alone, and the lactational amenorrhea method (LAM) are not acceptable methods of contraception. The female condom and the male condom should not be used together.

[0219] Treatment : REGN1979 will be administered by IV infusion at an initial dose of 1 mg during week 1, an intermediate dose of 20 mg at week 2, and a nominal dose of 80 mg or 160 mg during subsequent injections. For the initial dose, the intermediate dose and the first nominal dose (primary, secondary and tertiary doses, respectively), the treatment will be divided into 2 separate infusions, each for 4 hours on each of 2 days, preferably consecutive but not more than 3 days (for example, day 1 of week 1 and day 2 of week 1). Subsequent treatment (maximum weekly doses; eg, 320 mg) may be administered as a single infusion or as 2 separate infusions, and may be administered over 1 to 4 hours, depending on tolerance. Study treatment consisted of 12 doses of 1p/wk followed by 1p/2w of dosing for 86 weeks, for a total period of study drug administration of 98 weeks.

[0220] Endpoints : The primary endpoint of the study is ORR from the first dose to 194 weeks after the first dose according to the Lugano Classification of Response in Malignant Lymphoma (Cheson, 2014) and according to an independent centralized assessment, in patients with FL relapsed after at least at least two previous lines of systemic therapy, including an antibody to CD20 and an alkylating agent, or refractory to them. The 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) frequency of AE from the first dose to 194 weeks after the first dose, according to the Lugano classification, as assessed by an independent centralized assessment and assessment by a local investigator; (3) PFS from the first dose up to 194 weeks after the first dose, according to the Lugano classification, as assessed by an independent centralized assessment and assessment by a local investigator; (4) OS from first dose to 194 weeks after first dose; (5) TO from the first dose up to 194 weeks after the first dose, according to the Lugano classification, as determined by an independent centralized assessment and assessment by a local investigator; (6) CHKD from the first dose up to 194 weeks after the first dose, according to the Lugano classification, as assessed by an independent centralized assessment and assessment by a local investigator; (7) DCS from the first dose to 194 weeks after the first dose, according to the Lugano classification, as determined by an independent centralized assessment and assessment by a local investigator; (8) the incidence and severity of adverse events that occurred during treatment (AELI) from the first dose to 194 weeks after the first dose; and (9) change in patient outcome ratings, from first dose to 194 weeks after first dose, as measured by approved agents: EORTC QLQ-C30 and EQ-5D-3L.

[0221] Procedures and Evaluations : For all patients, the disease will be assessed radiologically using computed tomography (CT) or magnetic resonance imaging (MRI), and positron emission tomography imaging with ¹⁸ F-fluorodeoxyglucose (FDG-PET) injection. . Tumor response according to the Lugano classification criteria will be reviewed by an independent centralized radiological examination. A bone marrow aspirate, bone marrow biopsy, and lymph node and/or tumor biopsy will be taken and the samples will be evaluated histologically and may be used for other studies, including immunohistochemistry. Safety will be assessed by assessing vital signs, physical examination, Eastern Joint Oncology Group (ECOG) functional status score, electrocardiogram (ECG), AE rate, and providing information on concomitant medications. Laboratory tests include complete blood counts, blood chemistry, serum immunoglobulins G (IgG), serum pregnancy tests (if applicable), ferritin, and C-reactive protein (CRP). Blood samples will be taken for PK and anti-drug antibody (ADA) assessment. Peripheral blood samples will be taken to evaluate changes in biomarkers (eg, cytokine production, serum levels of pro-inflammatory cytokines, and changes in subpopulations and lymphocyte activation status). In addition, these samples will allow tumor or somatic genetic analysis of variations that influence the clinical course of the underlying disease or modulate the side effects of treatment. Quality of life assessment will be conducted using the EORTC QLQ-C30 and EQ-5D-3L self-administered questionnaires.

[0222] Statistical analysis design : This study is designed to evaluate the efficacy and safety of REGN1979 in patients with FL relapsed after at least two previous lines of systemic therapy, or refractory to them. Analysis of the primary efficacy endpoint will be performed after all patients have completed the 28-week study treatment period prior to evaluations and completed tumor response assessments, or dropped out of the study. Sample size rationale: A one-stage exact binomial design is adopted for the primary endpoint of the OER. Two-sided 95% confidence intervals for observed ORRs were calculated based on a sample size of 100. For 100 patients, if the observed ORR is at least 60%, 66%, 70%, and 75%, the lower limit of 95% CI would rule out an ORR of 49% , 55%, 60% and 65%, respectively; those. The ORR is significantly different from 49%, 55%, 60% and 65% as shown in Table 10 below.

Table 10 Two-tailed 95% exact confidence intervals for the observed ORR for a sample size of 100 patients

Number of respondents Observed MRO 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

With a sample size of 100 patients, if the true treatment effect of REGN1979 is 64%, 70%, 75%, or 80%, the probability that the observed lower bound of 95% CI would rule out 49%, 55%, 60%, or 65% is respectively 82%, 83%, 89% or 91%. The sample size will be further increased by 10% to account for patients who dropped out of the study prematurely. Thus, the total sample size will be 112 patients.

[0223] Statistical Methods : Patient population characteristics and baseline characteristics will be summarized using descriptive statistics. The primary efficacy endpoint is the OER according to the Lugano classification, based on an independent central assessment. ORRs will be summarized along with a two-sided 95% confidence interval. Patients not eligible for the best overall response score will be considered non-responders. Secondary efficacy endpoints of OER as determined by investigator assessment according to the Lugano classification, as well as RR and PCD rates as assessed by the local investigator and independent central assessment according to the Lugano classification, will be summarized together with a two-sided 95% confidence interval. Other secondary efficacy endpoints, including TO, DCH, PFS, and OS, will be summarized using the median and its 95% confidence interval using the Kaplan-Meier method according to the Lugano classification. Disease control rates will be summarized along with a two-sided confidence interval. Quality of life measured using approved tools: EORTC QLQ-C30 and EQ-5D-3L will be summarized using descriptive statistics. Observations and safety measures, including drug exposure, AEs, laboratory data, vital signs, and ECOG performance assessment, will be summarized and presented in tables and lists.

[0224] Interim Analysis : An interim analysis will be conducted after the first 50 patients complete tumor evaluation at 28 weeks or drop out of the study early. The ORR and corresponding 95% confidence interval will be summarized. Because the main purpose of this intermediate analysis is to point estimate of OER and to characterize the accuracy of the point estimate, no hypothesis testing is performed with this intermediate analysis. Thus Type I error correction is not applicable for this planned interim analysis. For other efficiency endpoints, a two-sided 95% confidence interval will also be presented.

[0225] Additional studies of the efficacy of REGN1979 may be included in this study or in additional studies, including (a) patients with follicular lymphoma (grades 1-3a) as third-line or higher (3L+) therapy, (b) patients with follicular lymphoma (grades 1-3a) eligible for full-dose chemotherapy as second or higher (2L+) line therapy, (c) patients with follicular lymphoma (grades 1-3a) not eligible for full-dose chemotherapy as 2L+ therapy , (d) patients with follicular lymphoma (grades 1-3a) previously untreated and eligible for full-dose chemoimmunotherapy, (e) patients with follicular lymphoma (grades 1-3a) previously untreated and not eligible for full-dose chemoimmunotherapy, (f) patients with follicular lymphoma (grades 1-3a) eligible for full-dose chemoimmunotherapy as first-line therapy (1L) instead of standard treatment, (g) patients with follicular lymphoma (grades 1-3a) not eligible for full-dose chemoimmunotherapy , as 1L therapy instead of standard treatment, (h) patients with follicular lymphoma (grades 1-3a) eligible for full-dose chemoimmunotherapy, as 2L+ therapy instead of standard treatment, (i) patients with follicular lymphoma (grades 1-3a), not eligible for full-dose chemoimmunotherapy, as 2L+ therapy instead of standard treatment, and/or (j) patients with follicular lymphoma in combination with standard treatment.

[0226] Additional studies of the efficacy of REGN1979 may be included in the present study or in additional studies, including (a) patients with diffuse large B-cell lymphoma (DLBCL), primary or transformed, as 3L+ therapy, (b) patients with DLBCL after failed CAR-T therapy, (c) DLBCL patients eligible for autologous HSCT as 2L+ therapy, (d) DLBCL patients not eligible for HSCT as 2L+ therapy, (e ) previously untreated DLBCL patients with poor molecular prognostic factors (B cells outside the germinal center, "double whammy" or "triple whammy") and eligible for full-dose chemoimmunotherapy, (f) previously untreated DLBCL patients , with poor molecular prognostic factors (B cells outside the germinal center, "double whammy" or "triple whammy") and not eligible for full-dose chemoimmunotherapy, (g) patients with DLBCL not previously treated with CAR-T, (h ) patients with DLBCL at a maximum weekly dose of 320 mg, (i) patients with DLBCL in combination with standard treatment, (j) patients with DLBCL for full-dose chemoimmunotherapy, as 1L therapy instead of standard treatment, (k) patients with DLBCL not eligible for full-dose chemoimmunotherapy, as 1L therapy instead of standard treatment, (l) DLBCL patients eligible for auto-HSCT as 2L+ therapy instead of standard treatment, and/or (m) DLBCL patients not eligible for auto-HSCT, as 2L+ therapy instead of standard treatment.

[0227] Additional studies of the efficacy of REGN1979 may be included in the present study or in additional studies, including (a) patients with mantle cell lymphoma (MCL) after treatment failure with a BTK inhibitor, as 2L+ therapy, (b) in patients with lymphoma marginal zone (LMZ), as 2L+ therapy; and/or (c) in patients with lymphoblastic lymphoma, lymphoplasmacytic lymphoma, Burkitt's lymphoma, or other B-NHL subtypes, as 2L+ therapy.

[0228] Additional studies of the efficacy of REGN1979 may be included in the present study or in additional studies, including (a) patients with CD20+ B cell malignancy who received a single dose of rituximab the day before the first dose of REGN1979. Only in this rituximab pre-dose cohort and extension cohort will a single dose of rituximab (375 mg/m ² ) be administered the day before the first dose of REGN1979 [i.e. on (-1) study day]. REGN1979 will be started on day 1 of week 1 and the treatment period for REGN1979 will be 9 months. Patients will receive up to 24 doses of REGN1979: 4 weekly doses during a 4-week induction period, followed by an additional 8 weekly doses and 12 doses given 1p/2wk over a 6-month maintenance period. In the first part of this pre-dose cohort, rituximab REGN1979 will be given with an 80 mg escalation dose. Once the optimal dosing regimen has been determined, one additional dose group of 6 patients at the 320 mg REGN1979 escalation dose will be evaluated at the optimal dosing regimen. Dose groups with increasing doses of REGN1979 from 80 to 320 mg can also be evaluated. A further 24 patients will then be evaluated at this optimal dosing schedule and optimal dose, and together with the 6 patients in the rituximab pre-dose arm treated with optimal dose, a total of 30 patients will be investigated for safety and tolerability.

[0229] In any of the standard treatment combination studies, the combination may include REGN1979 plus CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone), ICE (ifosfamide, carboplatin, and etoposide), Gem-Ox (gemcitabine and oxaliplatin), lenalidomide, or lenalidomide plus rituximab.

[0230] The scope of the present invention should not be limited to the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will be apparent to those skilled in the art from the foregoing description. Such modifications are intended to be covered by the appended claims.

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Ser Phe Gly Thr Leu Lys Lys Glu Gly Trp Arg Pro Arg Arg Thr Ile

405 410 415

Leu Phe Ala Ser Trp Asp Ala Glu Glu Phe Gly Leu Leu Gly Ser Thr

420 425 430

Glu Trp Ala Glu Glu Asn Ser Arg Leu Leu Gln Glu Arg Gly Val Ala

435 440 445

Tyr Ile Asn Ala Asp Ser Ser Ile Glu Gly Asn Tyr Thr Leu Arg Val

450 455 460

Asp Cys Thr Pro Leu Met Tyr Ser Leu Val His Asn Leu Thr Lys Glu

465 470 475 480

Leu Lys Ser Pro Asp Glu Gly Phe Glu Gly Lys Ser Leu Tyr Glu Ser

485 490 495

Trp Thr Lys Lys Ser Pro Ser Pro Glu Phe Ser Gly Met Pro Arg Ile

500 505 510

Ser Lys Leu Gly Ser Gly Asn Asp Phe Glu Val Phe Phe Gln Arg Leu

515 520 525

Gly Ile Ala Ser Gly Arg Ala Arg Tyr Thr Lys Asn Trp Glu Thr Asn

530 535 540

Lys Phe Ser Gly Tyr Pro Leu Tyr His Ser Val Tyr Glu Thr Tyr Glu

545 550 555 560

Leu Val Glu Lys Phe Tyr Asp Pro Met Phe Lys Tyr His Leu Thr Val

565 570 575

Ala Gln Val Arg Gly Gly Met Val Phe Glu Leu Ala Asn Ser Ile Val

580 585 590

Leu Pro Phe Asp Cys Arg Asp Tyr Ala Val Val Leu Arg Lys Tyr Ala

595 600 605

Asp Lys Ile Tyr Ser Ile Ser Met Lys His Pro Gln Glu Met Lys Thr

610 615 620

Tyr Ser Val Ser Phe Asp Ser Leu Phe Ser Ala Val Lys Asn Phe Thr

625 630 635 640

Glu Ile Ala Ser Lys Phe Ser Glu Arg Leu Gln Asp Phe Asp Lys Ser

645 650 655

Asn Pro Ile Val Leu Arg Met Met Asn Asp Gln Leu Met Phe Leu Glu

660 665 670

Arg Ala Phe Ile Asp Pro Leu Gly Leu Pro Asp Arg Pro Phe Tyr Arg

675 680 685

His Val Ile Tyr Ala Pro Ser Ser His Asn Lys Tyr Ala Gly Glu Ser

690 695 700

Phe Pro Gly Ile Tyr Asp Ala Leu Phe Asp Ile Glu Ser Lys Val Asp

705 710 715 720

Pro Ser Lys Ala Trp Gly Glu Val Lys Arg Gln Ile Tyr Val Ala Ala

725 730 735

Phe Thr Val Gln Ala Ala Ala Glu Thr Leu Ser Glu Val Ala

740 745 750

<210> 8

<211> 14507

<212> PRT

<213> Artificial sequence

<220>

<223> human protein MUC16 (CA-125)

<400> 8

Met Leu Lys Pro Ser Gly Leu Pro Gly Ser Ser Ser Pro Thr Arg Ser

1 5 10 15

Leu Met Thr Gly Ser Arg Ser Thr Lys Ala Thr Pro Glu Met Asp Ser

20 25 30

Gly Leu Thr Gly Ala Thr Leu Ser Pro Lys Thr Ser Thr Gly Ala Ile

35 40 45

Val Val Thr Glu His Thr Leu Pro Phe Thr Ser Pro Asp Lys Thr Leu

50 55 60

Ala Ser Pro Thr Ser Ser Val Val Gly Arg Thr Thr Gln Ser Leu Gly

65 70 75 80

Val Met Ser Ser Ala Leu Pro Glu Ser Thr Ser Arg Gly Met Thr His

85 90 95

Ser Glu Gln Arg Thr Ser Pro Ser Leu Ser Pro Gln Val Asn Gly Thr

100 105 110

Pro Ser Arg Asn Tyr Pro Ala Thr Ser Met Val Ser Gly Leu Ser Ser

115 120 125

Pro Arg Thr Arg Thr Ser Ser Thr Glu Gly Asn Phe Thr Lys Glu Ala

130 135 140

Ser Thr Tyr Thr Leu Thr Val Glu Thr Thr Ser Gly Pro Val Thr Glu

145 150 155 160

Lys Tyr Thr Val Pro Thr Glu Thr Ser Thr Thr Glu Gly Asp Ser Thr

165 170 175

Glu Thr Pro Trp Asp Thr Arg Tyr Ile Pro Val Lys Ile Thr Ser Pro

180 185 190

Met Lys Thr Phe Ala Asp Ser Thr Ala Ser Lys Glu Asn Ala Pro Val

195 200 205

Ser Met Thr Pro Ala Glu Thr Thr Val Thr Asp Ser His Thr Pro Gly

210 215 220

Arg Thr Asn Pro Ser Phe Gly Thr Leu Tyr Ser Ser Phe Leu Asp Leu

225 230 235 240

Ser Pro Lys Gly Thr Pro Asn Ser Arg Gly Glu Thr Ser Leu Glu Leu

245 250 255

Ile Leu Ser Thr Thr Gly Tyr Pro Phe Ser Ser Pro Glu Pro Gly Ser

260 265 270

Ala Gly His Ser Arg Ile Ser Thr Ser Ala Pro Leu Ser Ser Ser Ser Ala

275 280 285

Ser Val Leu Asp Asn Lys Ile Ser Glu Thr Ser Ile Phe Ser Gly Gln

290 295 300

Ser Leu Thr Ser Pro Leu Ser Pro Gly Val Pro Glu Ala Arg Ala Ser

305 310 315 320

Thr Met Pro Asn Ser Ala Ile Pro Phe Ser Met Thr Leu Ser Asn Ala

325 330 335

Glu Thr Ser Ala Glu Arg Val Arg Ser Thr Ile Ser Ser Leu Gly Thr

340 345 350

Pro Ser Ile Ser Thr Lys Gln Thr Ala Glu Thr Ile Leu Thr Phe His

355 360 365

Ala Phe Ala Glu Thr Met Asp Ile Pro Ser Thr His Ile Ala Lys Thr

370 375 380

Leu Ala Ser Glu Trp Leu Gly Ser Pro Gly Thr Leu Gly Gly Thr Ser

385 390 395 400

Thr Ser Ala Leu Thr Thr Thr Ser Pro Ser Thr Thr Leu Val Ser Glu

405 410 415

Glu Thr Asn Thr His His Ser Thr Ser Gly Lys Glu Thr Glu Gly Thr

420 425 430

Leu Asn Thr Ser Met Thr Pro Leu Glu Thr Ser Ala Pro Gly Glu Glu

435 440 445

Ser Glu Met Thr Ala Thr Leu Val Pro Thr Leu Gly Phe Thr Leu

450 455 460

Asp Ser Lys Ile Arg Ser Pro Ser Gln Val Ser Ser Ser His Pro Thr

465 470 475 480

Arg Glu Leu Arg Thr Thr Gly Ser Thr Ser Gly Arg Gln Ser Ser Ser

485 490 495

Thr Ala Ala His Gly Ser Ser Asp Ile Leu Arg Ala Thr Thr Ser Ser

500 505 510

Thr Ser Lys Ala Ser Ser Trp Thr Ser Glu Ser Thr Ala Gln Gln Phe

515 520 525

Ser Glu Pro Gln His Thr Gln Trp Val Glu Thr Ser Pro Ser Met Lys

530 535 540

Thr Glu Arg Pro Pro Ala Ser Thr Ser Val Ala Ala Pro Ile Thr Thr

545 550 555 560

Ser Val Pro Ser Val Val Ser Gly Phe Thr Thr Leu Lys Thr Ser Ser

565 570 575

Thr Lys Gly Ile Trp Leu Glu Glu Thr Ser Ala Asp Thr Leu Ile Gly

580 585 590

Glu Ser Thr Ala Gly Pro Thr Thr His Gln Phe Ala Val Pro Thr Gly

595 600 605

Ile Ser Met Thr Gly Gly Ser Ser Thr Arg Gly Ser Gln Gly Thr Thr

610 615 620

His Leu Leu Thr Arg Ala Thr Ala Ser Ser Glu Thr Ser Ala Asp Leu

625 630 635 640

Thr Leu Ala Thr Asn Gly Val Pro Val Ser Val Ser Pro Ala Val Ser

645 650 655

Lys Thr Ala Ala Gly Ser Ser Pro Pro Gly Gly Thr Lys Pro Ser Tyr

660 665 670

Thr Met Val Ser Ser Val Ile Pro Glu Thr Ser Ser Leu Gln Ser Ser

675 680 685

Ala Phe Arg Glu Gly Thr Ser Leu Gly Leu Thr Pro Leu Asn Thr Arg

690 695 700

His Pro Phe Ser Ser Pro Glu Pro Asp Ser Ala Gly His Thr Lys Ile

705 710 715 720

Ser Thr Ser Ile Pro Leu Leu Ser Ser Ala Ser Val Leu Glu Asp Lys

725 730 735

Val Ser Ala Thr Ser Thr Phe Ser His His Lys Ala Thr Ser Ser Ile

740 745 750

Thr Thr Gly Thr Pro Glu Ile Ser Thr Lys Thr Lys Pro Ser Ser Ala

755 760 765

Val Leu Ser Ser Met Thr Leu Ser Asn Ala Ala Thr Ser Pro Glu Arg

770 775 780

Val Arg Asn Ala Thr Ser Pro Leu Thr His Pro Ser Pro Ser Gly Glu

785 790 795 800

Glu Thr Ala Gly Ser Val Leu Thr Leu Ser Thr Ser Ala Glu Thr Thr

805 810 815

Asp Ser Pro Asn Ile His Pro Thr Gly Thr Leu Thr Ser Glu Ser Ser

820 825 830

Glu Ser Pro Ser Thr Leu Ser Leu Pro Ser Val Ser Gly Val Lys Thr

835 840 845

Thr Phe Ser Ser Ser Thr Pro Ser Thr His Leu Phe Thr Ser Gly Glu

850 855 860

Glu Thr Glu Glu Thr Ser Asn Pro Ser Val Ser Gln Pro Glu Thr Ser

865 870 875 880

Val Ser Arg Val Arg Thr Thr Leu Ala Ser Thr Ser Val Pro Thr Pro

885 890 895

Val Phe Pro Thr Met Asp Thr Trp Pro Thr Arg Ser Ala Gln Phe Ser

900 905 910

Ser Ser His Leu Val Ser Glu Leu Arg Ala Thr Ser Ser Thr Ser Val

915 920 925

Thr Asn Ser Thr Gly Ser Ala Leu Pro Lys Ile Ser His Leu Thr Gly

930 935 940

Thr Ala Thr Met Ser Gln Thr Asn Arg Asp Thr Phe Asn Asp Ser Ala

945 950 955 960

Ala Pro Gln Ser Thr Thr Trp Pro Glu Thr Ser Pro Arg Phe Lys Thr

965 970 975

Gly Leu Pro Ser Ala Thr Thr Thr Val Ser Thr Ser Ala Thr Ser Leu

980 985 990

Ser Ala Thr Val Met Val Ser Lys Phe Thr Ser Pro Ala Thr Ser Ser

995 1000 1005

Met Glu Ala Thr Ser Ile Arg Glu Pro Ser Thr Thr Ile Leu Thr

1010 1015 1020

Thr Glu Thr Thr Asn Gly Pro Gly Ser Met Ala Val Ala Ser Thr

1025 1030 1035

Asn Ile Pro Ile Gly Lys Gly Tyr Ile Thr Glu Gly Arg Leu Asp

1040 1045 1050

Thr Ser His Leu Pro Ile Gly Thr Thr Ala Ser Ser Glu Thr Ser

1055 1060 1065

Met Asp Phe Thr Met Ala Lys Glu Ser Val Ser Met Ser Val Ser

1070 1075 1080

Pro Ser Gln Ser Met Asp Ala Ala Gly Ser Ser Thr Pro Gly Arg

1085 1090 1095

Thr Ser Gln Phe Val Asp Thr Phe Ser Asp Asp Val Tyr His Leu

1100 1105 1110

Thr Ser Arg Glu Ile Thr Ile Pro Arg Asp Gly Thr Ser Ser Ala

1115 1120 1125

Leu Thr Pro Gln Met Thr Ala Thr His Pro Pro Ser Pro Asp Pro

1130 1135 1140

Gly Ser Ala Arg Ser Thr Trp Leu Gly Ile Leu Ser Ser Ser Pro

1145 1150 1155

Ser Ser Pro Thr Pro Lys Val Thr Met Ser Ser Thr Phe Ser Thr

1160 1165 1170

Gln Arg Val Thr Thr Ser Met Ile Met Asp Thr Val Glu Thr Ser

1175 1180 1185

Arg Trp Asn Met Pro Asn Leu Pro Ser Thr Thr Ser Leu Thr Pro

1190 1195 1200

Ser Asn Ile Pro Thr Ser Gly Ala Ile Gly Lys Ser Thr Leu Val

1205 1210 1215

Pro Leu Asp Thr Pro Ser Pro Ala Thr Ser Leu Glu Ala Ser Glu

1220 1225 1230

Gly Gly Leu Pro Thr Leu Ser Thr Tyr Pro Glu Ser Thr Asn Thr

1235 1240 1245

Pro Ser Ile His Leu Gly Ala His Ala Ser Ser Glu Ser Pro Ser

1250 1255 1260

Thr Ile Lys Leu Thr Met Ala Ser Val Val Lys Pro Gly Ser Tyr

1265 1270 1275

Thr Pro Leu Thr Phe Pro Ser Ile Glu Thr His Ile His Val Ser

1280 1285 1290

Thr Ala Arg Met Ala Tyr Ser Ser Gly Ser Ser Pro Glu Met Thr

1295 1300 1305

Ala Pro Gly Glu Thr Asn Thr Gly Ser Thr Trp Asp Pro Thr Thr

1310 1315 1320

Tyr Ile Thr Thr Thr Asp Pro Lys Asp Thr Ser Ser Ala Gln Val

1325 1330 1335

Ser Thr Pro His Ser Val Arg Thr Leu Arg Thr Thr Glu Asn His

1340 1345 1350

Pro Lys Thr Glu Ser Ala Thr Pro Ala Ala Tyr Ser Gly Ser Pro

1355 1360 1365

Lys Ile Ser Ser Ser Pro Asn Leu Thr Ser Pro Ala Thr Lys Ala

1370 1375 1380

Trp Thr Ile Thr Asp Thr Thr Glu His Ser Thr Gln Leu His Tyr

1385 1390 1395

Thr Lys Leu Ala Glu Lys Ser Ser Gly Phe Glu Thr Gln Ser Ala

1400 1405 1410

Pro Gly Pro Val Ser Val Val Ile Pro Thr Ser Pro Thr Ile Gly

1415 1420 1425

Ser Ser Thr Leu Glu Leu Thr Ser Asp Val Pro Gly Glu Pro Leu

1430 1435 1440

Val Leu Ala Pro Ser Glu Gln Thr Thr Ile Thr Leu Pro Met Ala

1445 1450 1455

Thr Trp Leu Ser Thr Ser Leu Thr Glu Glu Met Ala Ser Thr Asp

1460 1465 1470

Leu Asp Ile Ser Ser Pro Ser Ser Pro Met Ser Thr Phe Ala Ile

1475 1480 1485

Phe Pro Pro Met Ser Thr Pro Ser His Glu Leu Ser Lys Ser Glu

1490 1495 1500

Ala Asp Thr Ser Ala Ile Arg Asn Thr Asp Ser Thr Thr Leu Asp

1505 1510 1515

Gln His Leu Gly Ile Arg Ser Leu Gly Arg Thr Gly Asp Leu Thr

1520 1525 1530

Thr Val Pro Ile Thr Pro Leu Thr Thr Thr Trp Thr Ser Val Ile

1535 1540 1545

Glu His Ser Thr Gln Ala Gln Asp Thr Leu Ser Ala Thr Met Ser

1550 1555 1560

Pro Thr His Val Thr Gln Ser Leu Lys Asp Gln Thr Ser Ile Pro

1565 1570 1575

Ala Ser Ala Ser Pro Ser His Leu Thr Glu Val Tyr Pro Glu Leu

1580 1585 1590

Gly Thr Gln Gly Arg Ser Ser Ser Glu Ala Thr Thr Phe Trp Lys

1595 1600 1605

Pro Ser Thr Asp Thr Leu Ser Arg Glu Ile Glu Thr Gly Pro Thr

1610 1615 1620

Asn Ile Gln Ser Thr Pro Met Asp Asn Thr Thr Thr Gly Ser

1625 1630 1635

Ser Ser Ser Gly Val Thr Leu Gly Ile Ala His Leu Pro Ile Gly

1640 1645 1650

Thr Ser Ser Pro Ala Glu Thr Ser Thr Asn Met Ala Leu Glu Arg

1655 1660 1665

Arg Ser Ser Thr Ala Thr Val Ser Met Ala Gly Thr Met Gly Leu

1670 1675 1680

Leu Val Thr Ser Ala Pro Gly Arg Ser Ile Ser Gln Ser Leu Gly

1685 1690 1695

Arg Val Ser Ser Val Leu Ser Glu Ser Thr Thr Glu Gly Val Thr

1700 1705 1710

Asp Ser Ser Lys Gly Ser Ser Pro Arg Leu Asn Thr Gln Gly Asn

1715 1720 1725

Thr Ala Leu Ser Ser Ser Leu Glu Pro Ser Tyr Ala Glu Gly Ser

1730 1735 1740

Gln Met Ser Thr Ser Ile Pro Leu Thr Ser Ser Pro Thr Thr Pro

1745 1750 1755

Asp Val Glu Phe Ile Gly Gly Ser Thr Phe Trp Thr Lys Glu Val

1760 1765 1770

Thr Thr Val Met Thr Ser Asp Ile Ser Lys Ser Ser Ala Arg Thr

1775 1780 1785

Glu Ser Ser Ser Ala Thr Leu Met Ser Thr Ala Leu Gly Ser Thr

1790 1795 1800

Glu Asn Thr Gly Lys Glu Lys Leu Arg Thr Ala Ser Met Asp Leu

1805 1810 1815

Pro Ser Pro Thr Pro Ser Met Glu Val Thr Pro Trp Ile Ser Leu

1820 1825 1830

Thr Leu Ser Asn Ala Pro Asn Thr Thr Asp Ser Leu Asp Leu Ser

1835 1840 1845

His Gly Val His Thr Ser Ser Ala Gly Thr Leu Ala Thr Asp Arg

1850 1855 1860

Ser Leu Asn Thr Gly Val Thr Arg Ala Ser Arg Leu Glu Asn Gly

1865 1870 1875

Ser Asp Thr Ser Ser Lys Ser Leu Ser Met Gly Asn Ser Thr His

1880 1885 1890

Thr Ser Met Thr Tyr Thr Glu Lys Ser Glu Val Ser Ser Ser Ile

1895 1900 1905

His Pro Arg Pro Glu Thr Ser Ala Pro Gly Ala Glu Thr Thr Leu

1910 1915 1920

Thr Ser Thr Pro Gly Asn Arg Ala Ile Ser Leu Thr Leu Pro Phe

1925 1930 1935

Ser Ser Ile Pro Val Glu Glu Val Ile Ser Thr Gly Ile Thr Ser

1940 1945 1950

Gly Pro Asp Ile Asn Ser Ala Pro Met Thr His Ser Pro Ile Thr

1955 1960 1965

Pro Pro Thr Ile Val Trp Thr Ser Thr Gly Thr Ile Glu Gln Ser

1970 1975 1980

Thr Gln Pro Leu His Ala Val Ser Ser Glu Lys Val Ser Val Gln

1985 1990 1995

Thr Gln Ser Thr Pro Tyr Val Asn Ser Val Ala Val Ser Ala Ser

2000 2005 2010

Pro Thr His Glu Asn Ser Val Ser Ser Gly Ser Ser Thr Ser Ser

2015 2020 2025

Pro Tyr Ser Ser Ala Ser Leu Glu Ser Leu Asp Ser Thr Ile Ser

2030 2035 2040

Arg Arg Asn Ala Ile Thr Ser Trp Leu Trp Asp Leu Thr Thr Ser

2045 2050 2055

Leu Pro Thr Thr Thr Trp Pro Ser Thr Ser Leu Ser Glu Ala Leu

2060 2065 2070

Ser Ser Gly His Ser Gly Val Ser Asn Pro Ser Ser Thr Thr Thr

2075 2080 2085

Glu Phe Pro Leu Phe Ser Ala Ala Ser Thr Ser Ala Ala Lys Gln

2090 2095 2100

Arg Asn Pro Glu Thr Glu Thr His Gly Pro Gln Asn Thr Ala Ala

2105 2110 2115

Ser Thr Leu Asn Thr Asp Ala Ser Ser Val Thr Gly Leu Ser Glu

2120 2125 2130

Thr Pro Val Gly Ala Ser Ile Ser Ser Glu Val Pro Leu Pro Met

2135 2140 2145

Ala Ile Thr Ser Arg Ser Asp Val Ser Gly Leu Thr Ser Glu Ser

2150 2155 2160

Thr Ala Asn Pro Ser Leu Gly Thr Ala Ser Ser Ala Gly Thr Lys

2165 2170 2175

Leu Thr Arg Thr Ile Ser Leu Pro Thr Ser Glu Ser Leu Val Ser

2180 2185 2190

Phe Arg Met Asn Lys Asp Pro Trp Thr Val Ser Ile Pro Leu Gly

2195 2200 2205

Ser His Pro Thr Thr Asn Thr Glu Thr Ser Ile Pro Val Asn Ser

2210 2215 2220

Ala Gly Pro Pro Gly Leu Ser Thr Val Ala Ser Asp Val Ile Asp

2225 2230 2235

Thr Pro Ser Asp Gly Ala Glu Ser Ile Pro Thr Val Ser Phe Ser

2240 2245 2250

Pro Ser Pro Asp Thr Glu Val Thr Thr Ile Ser His Phe Pro Glu

2255 2260 2265

Lys Thr Thr His Ser Phe Arg Thr Ile Ser Ser Leu Thr His Glu

2270 2275 2280

Leu Thr Ser Arg Val Thr Pro Ile Pro Gly Asp Trp Met Ser Ser

2285 2290 2295

Ala Met Ser Thr Lys Pro Thr Gly Ala Ser Pro Ser Ile Thr Leu

2300 2305 2310

Gly Glu Arg Arg Thr Ile Thr Ser Ala Ala Pro Thr Thr Ser Pro

2315 2320 2325

Ile Val Leu Thr Ala Ser Phe Thr Glu Thr Ser Thr Val Ser Leu

2330 2335 2340

Asp Asn Glu Thr Thr Val Lys Thr Ser Asp Ile Leu Asp Ala Arg

2345 2350 2355

Lys Thr Asn Glu Leu Pro Ser Asp Ser Ser Ser Ser Ser Asp Leu

2360 2365 2370

Ile Asn Thr Ser Ile Ala Ser Ser Thr Met Asp Val Thr Lys Thr

2375 2380 2385

Ala Ser Ile Ser Pro Thr Ser Ile Ser Gly Met Thr Ala Ser Ser

2390 2395 2400

Ser Pro Ser Leu Phe Ser Ser Asp Arg Pro Gln Val Pro Thr Ser

2405 2410 2415

Thr Thr Glu Thr Asn Thr Ala Thr Ser Pro Ser Val Ser Ser Asn

2420 2425 2430

Thr Tyr Ser Leu Asp Gly Gly Ser Asn Val Gly Gly Thr Pro Ser

2435 2440 2445

Thr Leu Pro Pro Phe Thr Ile Thr His Pro Val Glu Thr Ser Ser

2450 2455 2460

Ala Leu Leu Ala Trp Ser Arg Pro Val Arg Thr Phe Ser Thr Met

2465 2470 2475

Val Ser Thr Asp Thr Ala Ser Gly Glu Asn Pro Thr Ser Ser Asn

2480 2485 2490

Ser Val Val Thr Ser Val Pro Ala Pro Gly Thr Trp Thr Ser Val

2495 2500 2505

Gly Ser Thr Thr Asp Leu Pro Ala Met Gly Phe Leu Lys Thr Ser

2510 2515 2520

Pro Ala Gly Glu Ala His Ser Leu Leu Ala Ser Thr Ile Glu Pro

2525 2530 2535

Ala Thr Ala Phe Thr Pro His Leu Ser Ala Ala Val Val Thr Gly

2540 2545 2550

Ser Ser Ala Thr Ser Glu Ala Ser Leu Leu Thr Thr Ser Glu Ser

2555 2560 2565

Lys Ala Ile His Ser Ser Pro Gln Thr Pro Thr Thr Pro Thr Ser

2570 2575 2580

Gly Ala Asn Trp Glu Thr Ser Ala Thr Pro Glu Ser Leu Leu Val

2585 2590 2595

Val Thr Glu Thr Ser Asp Thr Thr Leu Thr Ser Lys Ile Leu Val

2600 2605 2610

Thr Asp Thr Ile Leu Phe Ser Thr Val Ser Thr Pro Pro Ser Lys

2615 2620 2625

Phe Pro Ser Thr Gly Thr Leu Ser Gly Ala Ser Phe Pro Thr Leu

2630 2635 2640

Leu Pro Asp Thr Pro Ala Ile Pro Leu Thr Ala Thr Glu Pro Thr

2645 2650 2655

Ser Ser Leu Ala Thr Ser Phe Asp Ser Thr Pro Leu Val Thr Ile

2660 2665 2670

Ala Ser Asp Ser Leu Gly Thr Val Pro Glu Thr Thr Leu Thr Met

2675 2680 2685

Ser Glu Thr Ser Asn Gly Asp Ala Leu Val Leu Lys Thr Val Ser

2690 2695 2700

Asn Pro Asp Arg Ser Ile Pro Gly Ile Thr Ile Gln Gly Val Thr

2705 2710 2715

Glu Ser Pro Leu His Pro Ser Ser Thr Ser Pro Ser Lys Ile Val

2720 2725 2730

Ala Pro Arg Asn Thr Thr Tyr Glu Gly Ser Ile Thr Val Ala Leu

2735 2740 2745

Ser Thr Leu Pro Ala Gly Thr Thr Gly Ser Leu Val Phe Ser Gln

2750 2755 2760

Ser Ser Glu Asn Ser Glu Thr Thr Ala Leu Val Asp Ser Ser Ala

2765 2770 2775

Gly Leu Glu Arg Ala Ser Val Met Pro Leu Thr Thr Gly Ser Gln

2780 2785 2790

Gly Met Ala Ser Ser Gly Gly Ile Arg Ser Gly Ser Thr His Ser

2795 2800 2805

Thr Gly Thr Lys Thr Phe Ser Ser Leu Pro Leu Thr Met Asn Pro

2810 2815 2820

Gly Glu Val Thr Ala Met Ser Glu Ile Thr Asn Arg Leu Thr

2825 2830 2835

Ala Thr Gln Ser Thr Ala Pro Lys Gly Ile Pro Val Lys Pro Thr

2840 2845 2850

Ser Ala Glu Ser Gly Leu Leu Thr Pro Val Ser Ala Ser Ser Ser

2855 2860 2865

Pro Ser Lys Ala Phe Ala Ser Leu Thr Thr Ala Pro Pro Thr Trp

2870 2875 2880

Gly Ile Pro Gln Ser Thr Leu Thr Phe Glu Phe Ser Glu Val Pro

2885 2890 2895

Ser Leu Asp Thr Lys Ser Ala Ser Leu Pro Thr Pro Gly Gln Ser

2900 2905 2910

Leu Asn Thr Ile Pro Asp Ser Asp Ala Ser Thr Ala Ser Ser Ser

2915 2920 2925

Leu Ser Lys Ser Pro Glu Lys Asn Pro Arg Ala Arg Met Met Thr

2930 2935 2940

Ser Thr Lys Ala Ile Ser Ala Ser Ser Phe Gln Ser Thr Gly Phe

2945 2950 2955

Thr Glu Thr Pro Glu Gly Ser Ala Ser Pro Ser Met Ala Gly His

2960 2965 2970

Glu Pro Arg Val Pro Thr Ser Gly Thr Gly Asp Pro Arg Tyr Ala

2975 2980 2985

Ser Glu Ser Met Ser Tyr Pro Asp Pro Ser Lys Ala Ser Ser Ala

2990 2995 3000

Met Thr Ser Thr Ser Leu Ala Ser Lys Leu Thr Thr Leu Phe Ser

3005 3010 3015

Thr Gly Gln Ala Ala Arg Ser Gly Ser Ser Ser Ser Pro Ile Ser

3020 3025 3030

Leu Ser Thr Glu Lys Glu Thr Ser Phe Leu Ser Pro Thr Ala Ser

3035 3040 3045

Thr Ser Arg Lys Thr Ser Leu Phe Leu Gly Pro Ser Met Ala Arg

3050 3055 3060

Gln Pro Asn Ile Leu Val His Leu Gln Thr Ser Ala Leu Thr Leu

3065 3070 3075

Ser Pro Thr Ser Thr Leu Asn Met Ser Gln Glu Glu Pro Pro Glu

3080 3085 3090

Leu Thr Ser Ser Gln Thr Ile Ala Glu Glu Glu Gly Thr Thr Ala

3095 3100 3105

Glu Thr Gln Thr Leu Thr Phe Thr Pro Ser Glu Thr Pro Thr Ser

3110 3115 3120

Leu Leu Pro Val Ser Ser Pro Thr Glu Pro Thr Ala Arg Arg Lys

3125 3130 3135

Ser Ser Pro Glu Thr Trp Ala Ser Ser Ile Ser Val Pro Ala Lys

3140 3145 3150

Thr Ser Leu Val Glu Thr Thr Asp Gly Thr Leu Val Thr Thr Ile

3155 3160 3165

Lys Met Ser Ser Gln Ala Ala Gln Gly Asn Ser Thr Trp Pro Ala

3170 3175 3180

Pro Ala Glu Glu Thr Gly Ser Ser Pro Ala Gly Thr Ser Pro Gly

3185 3190 3195

Ser Pro Glu Met Ser Thr Thr Leu Lys Ile Met Ser Ser Lys Glu

3200 3205 3210

Pro Ser Ile Ser Pro Glu Ile Arg Ser Thr Val Arg Asn Ser Pro

3215 3220 3225

Trp Lys Thr Pro Glu Thr Thr Val Pro Met Glu Thr Thr Val Glu

3230 3235 3240

Pro Val Thr Leu Gln Ser Thr Ala Leu Gly Ser Gly Ser Thr Ser

3245 3250 3255

Ile Ser His Leu Pro Thr Gly Thr Thr Ser Pro Thr Lys Ser Pro

3260 3265 3270

Thr Glu Asn Met Leu Ala Thr Glu Arg Val Ser Leu Ser Pro Ser

3275 3280 3285

Pro Pro Glu Ala Trp Thr Asn Leu Tyr Ser Gly Thr Pro Gly Gly

3290 3295 3300

Thr Arg Gln Ser Leu Ala Thr Met Ser Ser Val Ser Leu Glu Ser

3305 3310 3315

Pro Thr Ala Arg Ser Ile Thr Gly Thr Gly Gln Gln Ser Ser Pro

3320 3325 3330

Glu Leu Val Ser Lys Thr Thr Gly Met Glu Phe Ser Met Trp His

3335 3340 3345

Gly Ser Thr Gly Gly Thr Thr Gly Asp Thr His Val Ser Leu Ser

3350 3355 3360

Thr Ser Ser Asn Ile Leu Glu Asp Pro Val Thr Ser Pro Asn Ser

3365 3370 3375

Val Ser Ser Leu Thr Asp Lys Ser Lys His Lys Thr Glu Thr Trp

3380 3385 3390

Val Ser Thr Thr Ala Ile Pro Ser Thr Val Leu Asn Asn Lys Ile

3395 3400 3405

Met Ala Ala Glu Gln Gln Thr Ser Arg Ser Val Asp Glu Ala Tyr

3410 3415 3420

Ser Ser Thr Ser Ser Trp Ser Asp Gln Thr Ser Gly Ser Asp Ile

3425 3430 3435

Thr Leu Gly Ala Ser Pro Asp Val Thr Asn Thr Leu Tyr Ile Thr

3440 3445 3450

Ser Thr Ala Gln Thr Thr Ser Leu Val Ser Leu Pro Ser Gly Asp

3455 3460 3465

Gln Gly Ile Thr Ser Leu Thr Asn Pro Ser Gly Gly Lys Thr Ser

3470 3475 3480

Ser Ala Ser Ser Val Thr Ser Pro Ser Ile Gly Leu Glu Thr Leu

3485 3490 3495

Arg Ala Asn Val Ser Ala Val Lys Ser Asp Ile Ala Pro Thr Ala

3500 3505 3510

Gly His Leu Ser Gln Thr Ser Ser Pro Ala Glu Val Ser Ile Leu

3515 3520 3525

Asp Val Thr Thr Ala Pro Thr Pro Gly Ile Ser Thr Thr Ile Thr

3530 3535 3540

Thr Met Gly Thr Asn Ser Ile Ser Thr Thr Thr Pro Asn Pro Glu

3545 3550 3555

Val Gly Met Ser Thr Met Asp Ser Thr Pro Ala Thr Glu Arg Arg

3560 3565 3570

Thr Thr Ser Thr Glu His Pro Ser Thr Trp Ser Ser Thr Ala Ala

3575 3580 3585

Ser Asp Ser Trp Thr Val Thr Asp Met Thr Ser Asn Leu Lys Val

3590 3595 3600

Ala Arg Ser Pro Gly Thr Ile Ser Thr Met His Thr Thr Ser Phe

3605 3610 3615

Leu Ala Ser Ser Thr Glu Leu Asp Ser Met Ser Thr Pro His Gly

3620 3625 3630

Arg Ile Thr Val Ile Gly Thr Ser Leu Val Thr Pro Ser Ser Asp

3635 3640 3645

Ala Ser Ala Val Lys Thr Glu Thr Ser Thr Ser Glu Arg Thr Leu

3650 3655 3660

Ser Pro Ser Asp Thr Thr Ala Ser Thr Pro Ile Ser Thr Phe Ser

3665 3670 3675

Arg Val Gln Arg Met Ser Ile Ser Val Pro Asp Ile Leu Ser Thr

3680 3685 3690

Ser Trp Thr Pro Ser Ser Thr Glu Ala Glu Asp Val Pro Val Ser

3695 3700 3705

Met Val Ser Thr Asp His Ala Ser Thr Lys Thr Asp Pro Asn Thr

3710 3715 3720

Pro Leu Ser Thr Phe Leu Phe Asp Ser Leu Ser Thr Leu Asp Trp

3725 3730 3735

Asp Thr Gly Arg Ser Leu Ser Ser Ala Thr Ala Thr Thr Ser Ala

3740 3745 3750

Pro Gln Gly Ala Thr Thr Pro Gln Glu Leu Thr Leu Glu Thr Met

3755 3760 3765

Ile Ser Pro Ala Thr Ser Gln Leu Pro Phe Ser Ile Gly His Ile

3770 3775 3780

Thr Ser Ala Val Thr Pro Ala Ala Met Ala Arg Ser Ser Gly Val

3785 3790 3795

Thr Phe Ser Arg Pro Asp Pro Thr Ser Lys Lys Ala Glu Gln Thr

3800 3805 3810

Ser Thr Gln Leu Pro Thr Thr Thr Ser Ala His Pro Gly Gln Val

3815 3820 3825

Pro Arg Ser Ala Ala Thr Thr Leu Asp Val Ile Pro His Thr Ala

3830 3835 3840

Lys Thr Pro Asp Ala Thr Phe Gln Arg Gln Gly Gln Thr Ala Leu

3845 3850 3855

Thr Thr Glu Ala Arg Ala Thr Ser Asp Ser Trp Asn Glu Lys Glu

3860 3865 3870

Lys Ser Thr Pro Ser Ala Pro Trp Ile Thr Glu Met Met Asn Ser

3875 3880 3885

Val Ser Glu Asp Thr Ile Lys Glu Val Thr Ser Ser Ser Ser Ser Val

3890 3895 3900

Leu Arg Thr Leu Asn Thr Leu Asp Ile Asn Leu Glu Ser Gly Thr

3905 3910 3915

Thr Ser Ser Pro Ser Trp Lys Ser Ser Pro Tyr Glu Arg Ile Ala

3920 3925 3930

Pro Ser Glu Ser Thr Thr Asp Lys Glu Ala Ile His Pro Ser Thr

3935 3940 3945

Asn Thr Val Glu Thr Thr Gly Trp Val Thr Ser Ser Glu His Ala

3950 3955 3960

Ser His Ser Thr Ile Pro Ala His Ser Ala Ser Ser Lys Leu Thr

3965 3970 3975

Ser Pro Val Val Thr Thr Ser Thr Arg Glu Gln Ala Ile Val Ser

3980 3985 3990

Met Ser Thr Thr Thr Trp Pro Glu Ser Thr Arg Ala Arg Thr Glu

3995 4000 4005

Pro Asn Ser Phe Leu Thr Ile Glu Leu Arg Asp Val Ser Pro Tyr

4010 4015 4020

Met Asp Thr Ser Ser Thr Thr Gln Thr Ser Ile Ile Ser Ser Pro

4025 4030 4035

Gly Ser Thr Ala Ile Thr Lys Gly Pro Arg Thr Glu Ile Thr Ser

4040 4045 4050

Ser Lys Arg Ile Ser Ser Phe Leu Ala Gln Ser Met Arg Ser

4055 4060 4065

Ser Asp Ser Pro Ser Glu Ala Ile Thr Arg Leu Ser Asn Phe Pro

4070 4075 4080

Ala Met Thr Glu Ser Gly Gly Met Ile Leu Ala Met Gln Thr Ser

4085 4090 4095

Pro Pro Gly Ala Thr Ser Leu Ser Ala Pro Thr Leu Asp Thr Ser

4100 4105 4110

Ala Thr Ala Ser Trp Thr Gly Thr Pro Leu Ala Thr Thr Gln Arg

4115 4120 4125

Phe Thr Tyr Ser Glu Lys Thr Thr Leu Phe Ser Lys Gly Pro Glu

4130 4135 4140

Asp Thr Ser Gln Pro Ser Pro Ser Val Glu Glu Thr Ser Ser

4145 4150 4155

Ser Ser Ser Leu Val Pro Ile His Ala Thr Thr Ser Pro Ser Asn

4160 4165 4170

Ile Leu Leu Thr Ser Gln Gly His Ser Pro Ser Ser Thr Pro Pro

4175 4180 4185

Val Thr Ser Val Phe Leu Ser Glu Thr Ser Gly Leu Gly Lys Thr

4190 4195 4200

Thr Asp Met Ser Arg Ile Ser Leu Glu Pro Gly Thr Ser Leu Pro

4205 4210 4215

Pro Asn Leu Ser Ser Thr Ala Gly Glu Ala Leu Ser Thr Tyr Glu

4220 4225 4230

Ala Ser Arg Asp Thr Lys Ala Ile His His Ser Ala Asp Thr Ala

4235 4240 4245

Val Thr Asn Met Glu Ala Thr Ser Ser Glu Tyr Ser Pro Ile Pro

4250 4255 4260

Gly His Thr Lys Pro Ser Lys Ala Thr Ser Pro Leu Val Thr Ser

4265 4270 4275

His Ile Met Gly Asp Ile Thr Ser Ser Thr Ser Val Phe Gly Ser

4280 4285 4290

Ser Glu Thr Thr Glu Ile Glu Thr Val Ser Ser Val Asn Gln Gly

4295 4300 4305

Leu Gln Glu Arg Ser Thr Ser Gln Val Ala Ser Ser Ala Thr Glu

4310 4315 4320

Thr Ser Thr Val Ile Thr His Val Ser Ser Gly Asp Ala Thr Thr

4325 4330 4335

His Val Thr Lys Thr Gln Ala Thr Phe Ser Ser Gly Thr Ser Ile

4340 4345 4350

Ser Ser Pro His Gln Phe Ile Thr Ser Thr Asn Thr Phe Thr Asp

4355 4360 4365

Val Ser Thr Asn Pro Ser Thr Ser Leu Ile Met Thr Glu Ser Ser

4370 4375 4380

Gly Val Thr Ile Thr Thr Gln Thr Gly Pro Thr Gly Ala Ala Thr

4385 4390 4395

Gln Gly Pro Tyr Leu Leu Asp Thr Ser Thr Met Pro Tyr Leu Thr

4400 4405 4410

Glu Thr Pro Leu Ala Val Thr Pro Asp Phe Met Gln Ser Glu Lys

4415 4420 4425

Thr Thr Leu Ile Ser Lys Gly Pro Lys Asp Val Ser Trp Thr Ser

4430 4435 4440

Pro Pro Ser Val Ala Glu Thr Ser Tyr Pro Ser Ser Leu Thr Pro

4445 4450 4455

Phe Leu Val Thr Thr Ile Pro Pro Ala Thr Ser Thr Leu Gln Gly

4460 4465 4470

Gln His Thr Ser Ser Pro Val Ser Ala Thr Ser Val Leu Thr Ser

4475 4480 4485

Gly Leu Val Lys Thr Thr Asp Met Leu Asn Thr Ser Met Glu Pro

4490 4495 4500

Val Thr Asn Ser Pro Gln Asn Leu Asn Asn Pro Ser Asn Glu Ile

4505 4510 4515

Leu Ala Thr Leu Ala Ala Thr Thr Asp Ile Glu Thr Ile His Pro

4520 4525 4530

Ser Ile Asn Lys Ala Val Thr Asn Met Gly Thr Ala Ser Ser Ala

4535 4540 4545

His Val Leu His Ser Thr Leu Pro Val Ser Ser Glu Pro Ser Thr

4550 4555 4560

Ala Thr Ser Pro Met Val Pro Ala Ser Ser Met Gly Asp Ala Leu

4565 4570 4575

Ala Ser Ile Ser Ile Pro Gly Ser Glu Thr Thr Asp Ile Glu Gly

4580 4585 4590

Glu Pro Thr Ser Ser Leu Thr Ala Gly Arg Lys Glu Asn Ser Thr

4595 4600 4605

Leu Gln Glu Met Asn Ser Thr Thr Glu Ser Asn Ile Ile Leu Ser

4610 4615 4620

Asn Val Ser Val Gly Ala Ile Thr Glu Ala Thr Lys Met Glu Val

4625 4630 4635

Pro Ser Phe Asp Ala Thr Phe Ile Pro Thr Pro Ala Gln Ser Thr

4640 4645 4650

Lys Phe Pro Asp Ile Phe Ser Val Ala Ser Ser Arg Leu Ser Asn

4655 4660 4665

Ser Pro Pro Met Thr Ile Ser Thr His Met Thr Thr Thr Gln Thr

4670 4675 4680

Gly Ser Ser Gly Ala Thr Ser Lys Ile Pro Leu Ala Leu Asp Thr

4685 4690 4695

Ser Thr Leu Glu Thr Ser Ala Gly Thr Pro Ser Val Val Thr Glu

4700 4705 4710

Gly Phe Ala His Ser Lys Ile Thr Thr Ala Met Asn Asn Asp Val

4715 4720 4725

Lys Asp Val Ser Gln Thr Asn Pro Pro Phe Gln Asp Glu Ala Ser

4730 4735 4740

Ser Pro Ser Ser Gln Ala Pro Val Leu Val Thr Thr Leu Pro Ser

4745 4750 4755

Ser Val Ala Phe Thr Pro Gln Trp His Ser Thr Ser Ser Pro Val

4760 4765 4770

Ser Met Ser Ser Val Leu Thr Ser Ser Leu Val Lys Thr Ala Gly

4775 4780 4785

Lys Val Asp Thr Ser Leu Glu Thr Val Thr Ser Ser Pro Gln Ser

4790 4795 4800

Met Ser Asn Thr Leu Asp Asp Ile Ser Val Thr Ser Ala Ala Thr

4805 4810 4815

Thr Asp Ile Glu Thr Thr His Pro Ser Ile Asn Thr Val Val Thr

4820 4825 4830

Asn Val Gly Thr Thr Gly Ser Ala Phe Glu Ser His Ser Thr Val

4835 4840 4845

Ser Ala Tyr Pro Glu Pro Ser Lys Val Thr Ser Pro Asn Val Thr

4850 4855 4860

Thr Ser Thr Met Glu Asp Thr Thr Ile Ser Arg Ser Ile Pro Lys

4865 4870 4875

Ser Ser Lys Thr Thr Arg Thr Glu Thr Glu Thr Thr Ser Ser Leu

4880 4885 4890

Thr Pro Lys Leu Arg Glu Thr Ser Ile Ser Gln Glu Ile Thr Ser

4895 4900 4905

Ser Thr Glu Thr Ser Thr Val Pro Tyr Lys Glu Leu Thr Gly Ala

4910 4915 4920

Thr Thr Glu Val Ser Arg Thr Asp Val Thr Ser Ser Ser Ser Ser Thr

4925 4930 4935

Ser Phe Pro Gly Pro Asp Gln Ser Thr Val Ser Leu Asp Ile Ser

4940 4945 4950

Thr Glu Thr Asn Thr Arg Leu Ser Thr Ser Pro Ile Met Thr Glu

4955 4960 4965

Ser Ala Glu Ile Thr Ile Thr Thr Gln Thr Gly Pro His Gly Ala

4970 4975 4980

Thr Ser Gln Asp Thr Phe Thr Met Asp Pro Ser Asn Thr Thr Pro

4985 4990 4995

Gln Ala Gly Ile His Ser Ala Met Thr His Gly Phe Ser Gln Leu

5000 5005 5010

Asp Val Thr Thr Leu Met Ser Arg Ile Pro Gln Asp Val Ser Trp

5015 5020 5025

Thr Ser Pro Pro Ser Val Asp Lys Thr Ser Ser Pro Ser Ser Phe

5030 5035 5040

Leu Ser Ser Pro Ala Met Thr Pro Ser Leu Ile Ser Ser Thr

5045 5050 5055

Leu Pro Glu Asp Lys Leu Ser Ser Pro Met Thr Ser Leu Leu Thr

5060 5065 5070

Ser Gly Leu Val Lys Ile Thr Asp Ile Leu Arg Thr Arg Leu Glu

5075 5080 5085

Pro Val Thr Ser Ser Leu Pro Asn Phe Ser Ser Thr Ser Asp Lys

5090 5095 5100

Ile Leu Ala Thr Ser Lys Asp Ser Lys Asp Thr Lys Glu Ile Phe

5105 5110 5115

Pro Ser Ile Asn Thr Glu Glu Thr Asn Val Lys Ala Asn Asn Ser

5120 5125 5130

Gly His Glu Ser His Ser Pro Ala Leu Ala Asp Ser Glu Thr Pro

5135 5140 5145

Lys Ala Thr Thr Gln Met Val Ile Thr Thr Thr Val Gly Asp Pro

5150 5155 5160

Ala Pro Ser Thr Ser Met Pro Val His Gly Ser Ser Glu Thr Thr

5165 5170 5175

Asn Ile Lys Arg Glu Pro Thr Tyr Phe Leu Thr Pro Arg Leu Arg

5180 5185 5190

Glu Thr Ser Thr Ser Gln Glu Ser Ser Phe Pro Thr Asp Thr Ser

5195 5200 5205

Phe Leu Leu Ser Lys Val Pro Thr Gly Thr Ile Thr Glu Val Ser

5210 5215 5220

Ser Thr Gly Val Asn Ser Ser Ser Lys Ile Ser Thr Pro Asp His

5225 5230 5235

Asp Lys Ser Thr Val Pro Pro Asp Thr Phe Thr Gly Glu Ile Pro

5240 5245 5250

Arg Val Phe Thr Ser Ser Ile Lys Thr Lys Ser Ala Glu Met Thr

5255 5260 5265

Ile Thr Thr Gln Ala Ser Pro Pro Glu Ser Ala Ser His Ser Thr

5270 5275 5280

Leu Pro Leu Asp Thr Ser Thr Thr Leu Ser Gln Gly Gly Thr His

5285 5290 5295

Ser Thr Val Thr Gln Gly Phe Pro Tyr Ser Glu Val Thr Thr Leu

5300 5305 5310

Met Gly Met Gly Pro Gly Asn Val Ser Trp Met Thr Thr Pro Pro

5315 5320 5325

Val Glu Glu Thr Ser Ser Val Ser Ser Leu Met Ser Ser Pro Ala

5330 5335 5340

Met Thr Ser Pro Ser Pro Val Ser Ser Thr Ser Pro Gln Ser Ile

5345 5350 5355

Pro Ser Ser Pro Leu Pro Val Thr Ala Leu Pro Thr Ser Val Leu

5360 5365 5370

Val Thr Thr Thr Asp Val Leu Gly Thr Thr Ser Pro Glu Ser Val

5375 5380 5385

Thr Ser Ser Pro Pro Asn Leu Ser Ser Ile Thr His Glu Arg Pro

5390 5395 5400

Ala Thr Tyr Lys Asp Thr Ala His Thr Glu Ala Ala Met His His

5405 5410 5415

Ser Thr Asn Thr Ala Val Thr Asn Val Gly Thr Ser Gly Ser Gly

5420 5425 5430

His Lys Ser Gln Ser Ser Val Leu Ala Asp Ser Glu Thr Ser Lys

5435 5440 5445

Ala Thr Pro Leu Met Ser Thr Thr Ser Thr Leu Gly Asp Thr Ser

5450 5455 5460

Val Ser Thr Ser Thr Pro Asn Ile Ser Gln Thr Asn Gln Ile Gln

5465 5470 5475

Thr Glu Pro Thr Ala Ser Leu Ser Pro Arg Leu Arg Glu Ser Ser

5480 5485 5490

Thr Ser Glu Lys Thr Ser Ser Thr Thr Glu Thr Asn Thr Ala Phe

5495 5500 5505

Ser Tyr Val Pro Thr Gly Ala Ile Thr Gln Ala Ser Arg Thr Glu

5510 5515 5520

Ile Ser Ser Ser Arg Thr Ser Ile Ser Asp Leu Asp Arg Pro Thr

5525 5530 5535

Ile Ala Pro Asp Ile Ser Thr Gly Met Ile Thr Arg Leu Phe Thr

5540 5545 5550

Ser Pro Ile Met Thr Lys Ser Ala Glu Met Thr Val Thr Thr Gln

5555 5560 5565

Thr Thr Thr Pro Gly Ala Thr Ser Gln Gly Ile Leu Pro Trp Asp

5570 5575 5580

Thr Ser Thr Thr Leu Phe Gln Gly Gly Thr His Ser Thr Val Ser

5585 5590 5595

Gln Gly Phe Pro His Ser Glu Ile Thr Thr Leu Arg Ser Arg Thr

5600 5605 5610

Pro Gly Asp Val Ser Trp Met Thr Thr Pro Pro Val Glu Glu Thr

5615 5620 5625

Ser Ser Gly Phe Ser Leu Met Ser Pro Ser Met Thr Ser Pro Ser

5630 5635 5640

Pro Val Ser Ser Thr Ser Pro Glu Ser Ile Pro Ser Ser Pro Leu

5645 5650 5655

Pro Val Thr Ala Leu Leu Thr Ser Val Leu Val Thr Thr Asn

5660 5665 5670

Val Leu Gly Thr Thr Ser Pro Glu Pro Val Thr Ser Ser Pro Pro

5675 5680 5685

Asn Leu Ser Ser Pro Thr Gln Glu Arg Leu Thr Thr Tyr Lys Asp

5690 5695 5700

Thr Ala His Thr Glu Ala Met His Ala Ser Met His Thr Asn Thr

5705 5710 5715

Ala Val Ala Asn Val Gly Thr Ser Ile Ser Gly His Glu Ser Gln

5720 5725 5730

Ser Ser Val Pro Ala Asp Ser His Thr Ser Lys Ala Thr Ser Pro

5735 5740 5745

Met Gly Ile Thr Phe Ala Met Gly Asp Thr Ser Val Ser Thr Ser

5750 5755 5760

Thr Pro Ala Phe Phe Glu Thr Arg Ile Gln Thr Glu Ser Thr Ser

5765 5770 5775

Ser Leu Ile Pro Gly Leu Arg Asp Thr Arg Thr Ser Glu Glu Ile

5780 5785 5790

Asn Thr Val Thr Glu Thr Ser Thr Val Leu Ser Glu Val Pro Thr

5795 5800 5805

Thr Thr Thr Thr Glu Val Ser Arg Thr Glu Val Ile Thr Ser Ser

5810 5815 5820

Arg Thr Thr Ile Ser Gly Pro Asp His Ser Lys Met Ser Pro Tyr

5825 5830 5835

Ile Ser Thr Glu Thr Ile Thr Arg Leu Ser Thr Phe Pro Phe Val

5840 5845 5850

Thr Gly Ser Thr Glu Met Ala Ile Thr Asn Gln Thr Gly Pro Ile

5855 5860 5865

Gly Thr Ile Ser Gln Ala Thr Leu Thr Leu Asp Thr Ser Ser Thr

5870 5875 5880

Ala Ser Trp Glu Gly Thr His Ser Pro Val Thr Gln Arg Phe Pro

5885 5890 5895

His Ser Glu Glu Thr Thr Thr Met Ser Arg Ser Thr Lys Gly Val

5900 5905 5910

Ser Trp Gln Ser Pro Pro Ser Val Glu Glu Thr Ser Ser Pro Ser

5915 5920 5925

Ser Pro Val Pro Leu Pro Ala Ile Thr Ser His Ser Ser Ser Leu Tyr

5930 5935 5940

Ser Ala Val Ser Gly Ser Ser Pro Thr Ser Ala Leu Pro Val Thr

5945 5950 5955

Ser Leu Leu Thr Ser Gly Arg Arg Lys Thr Ile Asp Met Leu Asp

5960 5965 5970

Thr His Ser Glu Leu Val Thr Ser Ser Leu Pro Ser Ala Ser Ser

5975 5980 5985

Phe Ser Gly Glu Ile Leu Thr Ser Glu Ala Ser Thr Asn Thr Glu

5990 5995 6000

Thr Ile His Phe Ser Glu Asn Thr Ala Glu Thr Asn Met Gly Thr

6005 6010 6015

Thr Asn Ser Met His Lys Leu His Ser Ser Val Ser Ile His Ser

6020 6025 6030

Gln Pro Ser Gly His Thr Pro Pro Lys Val Thr Gly Ser Met Met

6035 6040 6045

Glu Asp Ala Ile Val Ser Thr Ser Thr Pro Gly Ser Pro Glu Thr

6050 6055 6060

Lys Asn Val Asp Arg Asp Ser Thr Ser Pro Leu Thr Pro Glu Leu

6065 6070 6075

Lys Glu Asp Ser Thr Ala Leu Val Met Asn Ser Thr Thr Glu Ser

6080 6085 6090

Asn Thr Val Phe Ser Ser Val Ser Leu Asp Ala Ala Thr Glu Val

6095 6100 6105

Ser Arg Ala Glu Val Thr Tyr Tyr Asp Pro Thr Phe Met Pro Ala

6110 6115 6120

Ser Ala Gln Ser Thr Lys Ser Pro Asp Ile Ser Pro Glu Ala Ser

6125 6130 6135

Ser Ser His Ser Asn Ser Pro Pro Leu Thr Ile Ser Thr His Lys

6140 6145 6150

Thr Ile Ala Thr Gln Thr Gly Pro Ser Gly Val Thr Ser Leu Gly

6155 6160 6165

Gln Leu Thr Leu Asp Thr Ser Thr Ile Ala Thr Ser Ala Gly Thr

6170 6175 6180

Pro Ser Ala Arg Thr Gln Asp Phe Val Asp Ser Glu Thr Thr Ser

6185 6190 6195

Val Met Asn Asn Asp Leu Asn Asp Val Leu Lys Thr Ser Pro Phe

6200 6205 6210

Ser Ala Glu Glu Ala Asn Ser Leu Ser Ser Gln Ala Pro Leu Leu

6215 6220 6225

Val Thr Thr Ser Pro Ser Pro Val Thr Ser Thr Leu Gln Glu His

6230 6235 6240

Ser Thr Ser Ser Leu Val Ser Val Thr Ser Val Pro Thr Pro Thr

6245 6250 6255

Leu Ala Lys Ile Thr Asp Met Asp Thr Asn Leu Glu Pro Val Thr

6260 6265 6270

Arg Ser Pro Gln Asn Leu Arg Asn Thr Leu Ala Thr Ser Glu Ala

6275 6280 6285

Thr Thr Asp Thr His Thr Met His Pro Ser Ile Asn Thr Ala Val

6290 6295 6300

Ala Asn Val Gly Thr Thr Ser Ser Pro Asn Glu Phe Tyr Phe Thr

6305 6310 6315

Val Ser Pro Asp Ser Asp Pro Tyr Lys Ala Thr Ser Ala Val Val

6320 6325 6330

Ile Thr Ser Thr Ser Gly Asp Ser Ile Val Ser Thr Ser Met Pro

6335 6340 6345

Arg Ser Ser Ala Met Lys Lys Ile Glu Ser Glu Thr Thr Phe Ser

6350 6355 6360

Leu Ile Phe Arg Leu Arg Glu Thr Ser Thr Ser Gln Lys Ile Gly

6365 6370 6375

Ser Ser Ser Asp Thr Ser Thr Val Phe Asp Lys Ala Phe Thr Ala

6380 6385 6390

Ala Thr Thr Glu Val Ser Arg Thr Glu Leu Thr Ser Ser Ser Arg

6395 6400 6405

Thr Ser Ile Gln Gly Thr Glu Lys Pro Thr Met Ser Pro Asp Thr

6410 6415 6420

Ser Thr Arg Ser Val Thr Met Leu Ser Thr Phe Ala Gly Leu Thr

6425 6430 6435

Lys Ser Glu Glu Arg Thr Ile Ala Thr Gln Thr Gly Pro His Arg

6440 6445 6450

Ala Thr Ser Gln Gly Thr Leu Thr Trp Asp Thr Ser Ile Thr Thr

6455 6460 6465

Ser Gln Ala Gly Thr His Ser Ala Met Thr His Gly Phe Ser Gln

6470 6475 6480

Leu Asp Leu Ser Thr Leu Thr Ser Arg Val Pro Glu Tyr Ile Ser

6485 6490 6495

Gly Thr Ser Pro Pro Ser Val Glu Lys Thr Ser Ser Ser Ser Ser

6500 6505 6510

Leu Leu Ser Leu Pro Ala Ile Thr Ser Pro Ser Pro Val Pro Thr

6515 6520 6525

Thr Leu Pro Glu Ser Arg Pro Ser Ser Pro Val His Leu Thr Ser

6530 6535 6540

Leu Pro Thr Ser Gly Leu Val Lys Thr Thr Asp Met Leu Ala Ser

6545 6550 6555

Val Ala Ser Leu Pro Pro Asn Leu Gly Ser Thr Ser His Lys Ile

6560 6565 6570

Pro Thr Thr Ser Glu Asp Ile Lys Asp Thr Glu Lys Met Tyr Pro

6575 6580 6585

Ser Thr Asn Ile Ala Val Thr Asn Val Gly Thr Thr Thr Ser Glu

6590 6595 6600

Lys Glu Ser Tyr Ser Ser Val Pro Ala Tyr Ser Glu Pro Pro Lys

6605 6610 6615

Val Thr Ser Pro Met Val Thr Ser Phe Asn Ile Arg Asp Thr Ile

6620 6625 6630

Val Ser Thr Ser Met Pro Gly Ser Ser Glu Ile Thr Arg Ile Glu

6635 6640 6645

Met Glu Ser Thr Phe Ser Leu Ala His Gly Leu Lys Gly Thr Ser

6650 6655 6660

Thr Ser Gln Asp Pro Ile Val Ser Thr Glu Lys Ser Ala Val Leu

6665 6670 6675

His Lys Leu Thr Thr Gly Ala Thr Glu Thr Ser Arg Thr Glu Val

6680 6685 6690

Ala Ser Ser Arg Arg Thr Ser Ile Pro Gly Pro Asp His Ser Thr

6695 6700 6705

Glu Ser Pro Asp Ile Ser Thr Glu Val Ile Pro Ser Leu Pro Ile

6710 6715 6720

Ser Leu Gly Ile Thr Glu Ser Ser Asn Met Thr Ile Ile Thr Arg

6725 6730 6735

Thr Gly Pro Pro Leu Gly Ser Thr Ser Gln Gly Thr Phe Thr Leu

6740 6745 6750

Asp Thr Pro Thr Thr Ser Ser Arg Ala Gly Thr His Ser Met Ala

6755 6760 6765

Thr Gln Glu Phe Pro His Ser Glu Met Thr Thr Val Met Asn Lys

6770 6775 6780

Asp Pro Glu Ile Leu Ser Trp Thr Ile Pro Ser Ile Glu Lys

6785 6790 6795

Thr Ser Phe Ser Ser Ser Leu Met Pro Ser Pro Ala Met Thr Ser

6800 6805 6810

Pro Pro Val Ser Ser Thr Leu Pro Lys Thr Ile His Thr Thr Pro

6815 6820 6825

Ser Pro Met Thr Ser Leu Leu Thr Pro Ser Leu Val Met Thr Thr

6830 6835 6840

Asp Thr Leu Gly Thr Ser Pro Glu Pro Thr Thr Ser Ser Pro Pro

6845 6850 6855

Asn Leu Ser Ser Thr Ser His Glu Ile Leu Thr Thr Asp Glu Asp

6860 6865 6870

Thr Thr Ala Ile Glu Ala Met His Pro Ser Thr Ser Thr Ala Ala

6875 6880 6885

Thr Asn Val Glu Thr Thr Ser Ser Gly His Gly Ser Gln Ser Ser

6890 6895 6900

Val Leu Ala Asp Ser Glu Lys Thr Lys Ala Thr Ala Pro Met Asp

6905 6910 6915

Thr Thr Ser Thr Met Gly His Thr Thr Val Ser Thr Ser Met Ser

6920 6925 6930

Val Ser Ser Glu Thr Thr Lys Ile Lys Arg Glu Ser Thr Tyr Ser

6935 6940 6945

Leu Thr Pro Gly Leu Arg Glu Thr Ser Ile Ser Gln Asn Ala Ser

6950 6955 6960

Phe Ser Thr Asp Thr Ser Ile Val Leu Ser Glu Val Pro Thr Gly

6965 6970 6975

Thr Thr Ala Glu Val Ser Arg Thr Glu Val Thr Ser Ser Gly Arg

6980 6985 6990

Thr Ser Ile Pro Gly Pro Ser Gln Ser Thr Val Leu Pro Glu Ile

6995 7000 7005

Ser Thr Arg Thr Met Thr Arg Leu Phe Ala Ser Pro Thr Met Thr

7010 7015 7020

Glu Ser Ala Glu Met Thr Ile Pro Thr Gln Thr Gly Pro Ser Gly

7025 7030 7035

Ser Thr Ser Gln Asp Thr Leu Thr Leu Asp Thr Ser Thr Thr Lys

7040 7045 7050

Ser Gln Ala Lys Thr His Ser Thr Leu Thr Gln Arg Phe Pro His

7055 7060 7065

Ser Glu Met Thr Thr Leu Met Ser Arg Gly Pro Gly Asp Met Ser

7070 7075 7080

Trp Gln Ser Ser Pro Ser Leu Glu Asn Pro Ser Ser Leu Pro Ser

7085 7090 7095

Leu Leu Ser Leu Pro Ala Thr Thr Ser Pro Pro Pro Ile Ser Ser

7100 7105 7110

Thr Leu Pro Val Thr Ile Ser Ser Ser Pro Leu Pro Val Thr Ser

7115 7120 7125

Leu Leu Thr Ser Ser Pro Val Thr Thr Thr Asp Met Leu His Thr

7130 7135 7140

Ser Pro Glu Leu Val Thr Ser Ser Pro Pro Lys Leu Ser His Thr

7145 7150 7155

Ser Asp Glu Arg Leu Thr Thr Gly Lys Asp Thr Thr Asn Thr Glu

7160 7165 7170

Ala Val His Pro Ser Thr Asn Thr Ala Ala Ser Asn Val Glu Ile

7175 7180 7185

Pro Ser Ser Gly His Glu Ser Pro Ser Ser Ala Leu Ala Asp Ser

7190 7195 7200

Glu Thr Ser Lys Ala Thr Ser Pro Met Phe Ile Thr Ser Thr Gln

7205 7210 7215

Glu Asp Thr Thr Val Ala Ile Ser Thr Pro His Phe Leu Glu Thr

7220 7225 7230

Ser Arg Ile Gln Lys Glu Ser Ile Ser Ser Ser Leu Ser Pro Lys Leu

7235 7240 7245

Arg Glu Thr Gly Ser Ser Val Glu Thr Ser Ser Ala Ile Glu Thr

7250 7255 7260

Ser Ala Val Leu Ser Glu Val Ser Ile Gly Ala Thr Thr Glu Ile

7265 7270 7275

Ser Arg Thr Glu Val Thr Ser Ser Ser Arg Thr Ser Ile Ser Gly

7280 7285 7290

Ser Ala Glu Ser Thr Met Leu Pro Glu Ile Ser Thr Thr Arg Lys

7295 7300 7305

Ile Ile Lys Phe Pro Thr Ser Pro Ile Leu Ala Glu Ser Ser Glu

7310 7315 7320

Met Thr Ile Lys Thr Gln Thr Ser Pro Pro Gly Ser Thr Ser Glu

7325 7330 7335

Ser Thr Phe Thr Leu Asp Thr Ser Thr Thr Pro Ser Leu Val Ile

7340 7345 7350

Thr His Ser Thr Met Thr Gln Arg Leu Pro His Ser Glu Ile Thr

7355 7360 7365

Thr Leu Val Ser Arg Gly Ala Gly Asp Val Pro Arg Pro Ser Ser

7370 7375 7380

Leu Pro Val Glu Glu Thr Ser Pro Pro Ser Ser Gln Leu Ser Leu

7385 7390 7395

Ser Ala Met Ile Ser Pro Ser Pro Val Ser Ser Thr Leu Pro Ala

7400 7405 7410

Ser Ser His Ser Ser Ser Ala Ser Val Thr Ser Leu Leu Thr Pro

7415 7420 7425

Gly Gln Val Lys Thr Thr Glu Val Leu Asp Ala Ser Ala Glu Pro

7430 7435 7440

Glu Thr Ser Ser Pro Pro Ser Leu Ser Ser Thr Ser Val Glu Ile

7445 7450 7455

Leu Ala Thr Ser Glu Val Thr Thr Asp Thr Glu Lys Ile His Pro

7460 7465 7470

Phe Ser Asn Thr Ala Val Thr Lys Val Gly Thr Ser Ser Ser Gly

7475 7480 7485

His Glu Ser Pro Ser Ser Val Leu Pro Asp Ser Glu Thr Thr Lys

7490 7495 7500

Ala Thr Ser Ala Met Gly Thr Ile Ser Ile Met Gly Asp Thr Ser

7505 7510 7515

Val Ser Thr Leu Thr Pro Ala Leu Ser Asn Thr Arg Lys Ile Gln

7520 7525 7530

Ser Glu Pro Ala Ser Ser Leu Thr Thr Arg Leu Arg Glu Thr Ser

7535 7540 7545

Thr Ser Glu Glu Thr Ser Leu Ala Thr Glu Ala Asn Thr Val Leu

7550 7555 7560

Ser Lys Val Ser Thr Gly Ala Thr Thr Glu Val Ser Arg Thr Glu

7565 7570 7575

Ala Ile Ser Phe Ser Arg Thr Ser Met Ser Gly Pro Glu Gln Ser

7580 7585 7590

Thr Met Ser Gln Asp Ile Ser Ile Gly Thr Ile Pro Arg Ile Ser

7595 7600 7605

Ala Ser Ser Val Leu Thr Glu Ser Ala Lys Met Thr Ile Thr Thr

7610 7615 7620

Gln Thr Gly Pro Ser Glu Ser Thr Leu Glu Ser Thr Leu Asn Leu

7625 7630 7635

Asn Thr Ala Thr Thr Pro Ser Trp Val Glu Thr His Ser Ile Val

7640 7645 7650

Ile Gln Gly Phe Pro His Pro Glu Met Thr Thr Ser Met Gly Arg

7655 7660 7665

Gly Pro Gly Gly Val Ser Trp Pro Ser Pro Pro Phe Val Lys Glu

7670 7675 7680

Thr Ser Pro Pro Ser Ser Pro Leu Ser Leu Pro Ala Val Thr Ser

7685 7690 7695

Pro His Pro Val Ser Thr Thr Phe Leu Ala His Ile Pro Pro Ser

7700 7705 7710

Pro Leu Pro Val Thr Ser Leu Leu Thr Ser Gly Pro Ala Thr Thr

7715 7720 7725

Thr Asp Ile Leu Gly Thr Ser Thr Glu Pro Gly Thr Ser Ser Ser

7730 7735 7740

Ser Ser Leu Ser Thr Thr Ser His Glu Arg Leu Thr Thr Tyr Lys

7745 7750 7755

Asp Thr Ala His Thr Glu Ala Val His Pro Ser Thr Asn Thr Gly

7760 7765 7770

Gly Thr Asn Val Ala Thr Thr Ser Ser Gly Tyr Lys Ser Gln Ser

7775 7780 7785

Ser Val Leu Ala Asp Ser Ser Pro Met Cys Thr Ser Thr Met

7790 7795 7800

Gly Asp Thr Ser Val Leu Thr Ser Thr Pro Ala Phe Leu Glu Thr

7805 7810 7815

Arg Arg Ile Gln Thr Glu Leu Ala Ser Ser Leu Thr Pro Gly Leu

7820 7825 7830

Arg Glu Ser Ser Gly Ser Glu Gly Thr Ser Ser Gly Thr Lys Met

7835 7840 7845

Ser Thr Val Leu Ser Lys Val Pro Thr Gly Ala Thr Thr Glu Ile

7850 7855 7860

Ser Lys Glu Asp Val Thr Ser Ile Pro Gly Pro Ala Gln Ser Thr

7865 7870 7875

Ile Ser Pro Asp Ile Ser Thr Arg Thr Val Ser Trp Phe Ser Thr

7880 7885 7890

Ser Pro Val Met Thr Glu Ser Ala Glu Ile Thr Met Asn Thr His

7895 7900 7905

Thr Ser Pro Leu Gly Ala Thr Thr Gln Gly Thr Ser Thr Leu Asp

7910 7915 7920

Thr Ser Ser Thr Thr Ser Leu Thr Met Thr His Ser Thr Ile Ser

7925 7930 7935

Gln Gly Phe Ser His Ser Gln Met Ser Thr Leu Met Arg Arg Gly

7940 7945 7950

Pro Glu Asp Val Ser Trp Met Ser Pro Pro Leu Leu Glu Lys Thr

7955 7960 7965

Arg Pro Ser Phe Ser Leu Met Ser Ser Pro Ala Thr Thr Ser Pro

7970 7975 7980

Ser Pro Val Ser Ser Thr Leu Pro Glu Ser Ile Ser Ser Ser Pro

7985 7990 7995

Leu Pro Val Thr Ser Leu Leu Thr Ser Gly Leu Ala Lys Thr Thr

8000 8005 8010

Asp Met Leu His Lys Ser Ser Glu Pro Val Thr Asn Ser Pro Ala

8015 8020 8025

Asn Leu Ser Ser Thr Ser Val Glu Ile Leu Ala Thr Ser Glu Val

8030 8035 8040

Thr Thr Asp Thr Glu Lys Thr His Pro Ser Ser Asn Arg Thr Val

8045 8050 8055

Thr Asp Val Gly Thr Ser Ser Ser Gly His Glu Ser Thr Ser Phe

8060 8065 8070

Val Leu Ala Asp Ser Gln Thr Ser Lys Val Thr Ser Pro Met Val

8075 8080 8085

Ile Thr Ser Thr Met Glu Asp Thr Ser Val Ser Thr Ser Thr Pro

8090 8095 8100

Gly Phe Phe Glu Thr Ser Arg Ile Gln Thr Glu Pro Thr Ser Ser

8105 8110 8115

Leu Thr Leu Gly Leu Arg Lys Thr Ser Ser Ser Glu Gly Thr Ser

8120 8125 8130

Leu Ala Thr Glu Met Ser Thr Val Leu Ser Gly Val Pro Thr Gly

8135 8140 8145

Ala Thr Ala Glu Val Ser Arg Thr Glu Val Thr Ser Ser Ser Arg

8150 8155 8160

Thr Ser Ile Ser Gly Phe Ala Gln Leu Thr Val Ser Pro Glu Thr

8165 8170 8175

Ser Thr Glu Thr Ile Thr Arg Leu Pro Thr Ser Ser Ser Ile Met Thr

8180 8185 8190

Glu Ser Ala Glu Met Met Ile Lys Thr Gln Thr Asp Pro Pro Gly

8195 8200 8205

Ser Thr Pro Glu Ser Thr His Thr Val Asp Ile Ser Thr Thr Pro

8210 8215 8220

Asn Trp Val Glu Thr His Ser Thr Val Thr Gln Arg Phe Ser His

8225 8230 8235

Ser Glu Met Thr Thr Leu Val Ser Arg Ser Pro Gly Asp Met Leu

8240 8245 8250

Trp Pro Ser Gln Ser Ser Val Glu Glu Thr Ser Ser Ala Ser Ser

8255 8260 8265

Leu Leu Ser Leu Pro Ala Thr Thr Ser Pro Ser Pro Val Ser Ser

8270 8275 8280

Thr Leu Val Glu Asp Phe Pro Ser Ala Ser Leu Pro Val Thr Ser

8285 8290 8295

Leu Leu Asn Pro Gly Leu Val Ile Thr Thr Asp Arg Met Gly Ile

8300 8305 8310

Ser Arg Glu Pro Gly Thr Ser Ser Thr Ser Asn Leu Ser Ser Thr

8315 8320 8325

Ser His Glu Arg Leu Thr Thr Leu Glu Asp Thr Val Asp Thr Glu

8330 8335 8340

Asp Met Gln Pro Ser Thr His Thr Ala Val Thr Asn Val Arg Thr

8345 8350 8355

Ser Ile Ser Gly His Glu Ser Gln Ser Ser Val Leu Ser Asp Ser

8360 8365 8370

Glu Thr Pro Lys Ala Thr Ser Pro Met Gly Thr Thr Tyr Thr Met

8375 8380 8385

Gly Glu Thr Ser Val Ser Ile Ser Thr Ser Asp Phe Phe Glu Thr

8390 8395 8400

Ser Arg Ile Gln Ile Glu Pro Thr Ser Ser Leu Thr Ser Gly Leu

8405 8410 8415

Arg Glu Thr Ser Ser Ser Glu Arg Ile Ser Ser Ala Thr Glu Gly

8420 8425 8430

Ser Thr Val Leu Ser Glu Val Pro Ser Gly Ala Thr Thr Glu Val

8435 8440 8445

Ser Arg Thr Glu Val Ile Ser Ser Arg Gly Thr Ser Met Ser Gly

8450 8455 8460

Pro Asp Gln Phe Thr Ile Ser Pro Asp Ile Ser Thr Glu Ala Ile

8465 8470 8475

Thr Arg Leu Ser Thr Ser Pro Ile Met Thr Glu Ser Ala Glu Ser

8480 8485 8490

Ala Ile Thr Ile Glu Thr Gly Ser Pro Gly Ala Thr Ser Glu Gly

8495 8500 8505

Thr Leu Thr Leu Asp Thr Ser Thr Thr Thr Phe Trp Ser Gly Thr

8510 8515 8520

His Ser Thr Ala Ser Pro Gly Phe Ser His Ser Glu Met Thr Thr

8525 8530 8535

Leu Met Ser Arg Thr Pro Gly Asp Val Pro Trp Pro Ser Leu Pro

8540 8545 8550

Ser Val Glu Glu Ala Ser Ser Val Ser Ser Ser Leu Ser Ser Pro

8555 8560 8565

Ala Met Thr Ser Thr Ser Phe Phe Ser Thr Leu Pro Glu Ser Ile

8570 8575 8580

Ser Ser Ser Pro His Pro Val Thr Ala Leu Leu Thr Leu Gly Pro

8585 8590 8595

Val Lys Thr Thr Asp Met Leu Arg Thr Ser Ser Glu Pro Glu Thr

8600 8605 8610

Ser Ser Pro Pro Asn Leu Ser Ser Thr Ser Ala Glu Ile Leu Ala

8615 8620 8625

Thr Ser Glu Val Thr Lys Asp Arg Glu Lys Ile His Pro Ser Ser

8630 8635 8640

Asn Thr Pro Val Val Asn Val Gly Thr Val Ile Tyr Lys His Leu

8645 8650 8655

Ser Pro Ser Ser Val Leu Ala Asp Leu Val Thr Thr Lys Pro Thr

8660 8665 8670

Ser Pro Met Ala Thr Thr Ser Thr Leu Gly Asn Thr Ser Val Ser

8675 8680 8685

Thr Ser Thr Pro Ala Phe Pro Glu Thr Met Met Thr Gln Pro Thr

8690 8695 8700

Ser Ser Leu Thr Ser Gly Leu Arg Glu Ile Ser Thr Ser Gln Glu

8705 8710 8715

Thr Ser Ser Ala Thr Glu Arg Ser Ala Ser Leu Ser Gly Met Pro

8720 8725 8730

Thr Gly Ala Thr Thr Lys Val Ser Arg Thr Glu Ala Leu Ser Leu

8735 8740 8745

Gly Arg Thr Ser Thr Pro Gly Pro Ala Gln Ser Thr Ile Ser Pro

8750 8755 8760

Glu Ile Ser Thr Glu Thr Ile Thr Arg Ile Ser Thr Pro Leu Thr

8765 8770 8775

Thr Thr Gly Ser Ala Glu Met Thr Ile Thr Pro Lys Thr Gly His

8780 8785 8790

Ser Gly Ala Ser Ser Gln Gly Thr Phe Thr Leu Asp Thr Ser Ser

8795 8800 8805

Arg Ala Ser Trp Pro Gly Thr His Ser Ala Ala Thr His Arg Ser

8810 8815 8820

Pro His Ser Gly Met Thr Thr Pro Met Ser Arg Gly Pro Glu Asp

8825 8830 8835

Val Ser Trp Pro Ser Arg Pro Ser Val Glu Lys Thr Ser Pro Pro

8840 8845 8850

Ser Ser Leu Val Ser Leu Ser Ala Val Thr Ser Pro Ser Pro Leu

8855 8860 8865

Tyr Ser Thr Pro Ser Glu Ser Ser His Ser Ser Pro Leu Arg Val

8870 8875 8880

Thr Ser Leu Phe Thr Pro Val Met Met Lys Thr Thr Asp Met Leu

8885 8890 8895

Asp Thr Ser Leu Glu Pro Val Thr Thr Ser Pro Pro Ser Met Asn

8900 8905 8910

Ile Thr Ser Asp Glu Ser Leu Ala Thr Ser Lys Ala Thr Met Glu

8915 8920 8925

Thr Glu Ala Ile Gln Leu Ser Glu Asn Thr Ala Val Thr Gln Met

8930 8935 8940

Gly Thr Ile Ser Ala Arg Gln Glu Phe Tyr Ser Ser Tyr Pro Gly

8945 8950 8955

Leu Pro Glu Pro Ser Lys Val Thr Ser Pro Val Val Thr Ser Ser

8960 8965 8970

Thr Ile Lys Asp Ile Val Ser Thr Thr Ile Pro Ala Ser Ser Glu

8975 8980 8985

Ile Thr Arg Ile Glu Met Glu Ser Thr Ser Thr Leu Thr Pro Thr

8990 8995 9000

Pro Arg Glu Thr Ser Thr Ser Gln Glu Ile His Ser Ala Thr Lys

9005 9010 9015

Pro Ser Thr Val Pro Tyr Lys Ala Leu Thr Ser Ala Thr Ile Glu

9020 9025 9030

Asp Ser Met Thr Gln Val Met Ser Ser Ser Arg Gly Pro Ser Pro

9035 9040 9045

Asp Gln Ser Thr Met Ser Gln Asp Ile Ser Thr Glu Val Ile Thr

9050 9055 9060

Arg Leu Ser Thr Ser Pro Ile Lys Thr Glu Ser Thr Glu Met Thr

9065 9070 9075

Ile Thr Thr Gln Thr Gly Ser Pro Gly Ala Thr Ser Arg Gly Thr

9080 9085 9090

Leu Thr Leu Asp Thr Ser Thr Thr Phe Met Ser Gly Thr His Ser

9095 9100 9105

Thr Ala Ser Gln Gly Phe Ser His Ser Gln Met Thr Ala Leu Met

9110 9115 9120

Ser Arg Thr Pro Gly Asp Val Pro Trp Leu Ser His Pro Ser Val

9125 9130 9135

Glu Glu Ala Ser Ser Ala Ser Phe Ser Leu Ser Ser Pro Val Met

9140 9145 9150

Thr Ser Ser Ser Pro Val Ser Ser Thr Leu Pro Asp Ser Ile His

9155 9160 9165

Ser Ser Ser Leu Pro Val Thr Ser Leu Leu Thr Ser Gly Leu Val

9170 9175 9180

Lys Thr Thr Glu Leu Leu Gly Thr Ser Ser Glu Pro Glu Thr Ser

9185 9190 9195

Ser Pro Pro Asn Leu Ser Ser Thr Ser Ala Glu Ile Leu Ala Ile

9200 9205 9210

Thr Glu Val Thr Thr Asp Thr Glu Lys Leu Glu Met Thr Asn Val

9215 9220 9225

Val Thr Ser Gly Tyr Thr His Glu Ser Pro Ser Ser Val Leu Ala

9230 9235 9240

Asp Ser Val Thr Thr Lys Ala Thr Ser Ser Met Gly Ile Thr Tyr

9245 9250 9255

Pro Thr Gly Asp Thr Asn Val Leu Thr Ser Thr Pro Ala Phe Ser

9260 9265 9270

Asp Thr Ser Arg Ile Gln Thr Lys Ser Lys Leu Ser Leu Thr Pro

9275 9280 9285

Gly Leu Met Glu Thr Ser Ile Ser Glu Glu Thr Ser Ser Ala Thr

9290 9295 9300

Glu Lys Ser Thr Val Leu Ser Ser Val Pro Thr Gly Ala Thr Thr

9305 9310 9315

Glu Val Ser Arg Thr Glu Ala Ile Ser Ser Ser Arg Thr Ser Ile

9320 9325 9330

Pro Gly Pro Ala Gln Ser Thr Met Ser Ser Asp Thr Ser Met Glu

9335 9340 9345

Thr Ile Thr Arg Ile Ser Thr Pro Leu Thr Arg Lys Glu Ser Thr

9350 9355 9360

Asp Met Ala Ile Thr Pro Lys Thr Gly Pro Ser Gly Ala Thr Ser

9365 9370 9375

Gln Gly Thr Phe Thr Leu Asp Ser Ser Ser Thr Ala Ser Trp Pro

9380 9385 9390

Gly Thr His Ser Ala Thr Thr Gln Arg Phe Pro Gln Ser Val Val

9395 9400 9405

Thr Thr Pro Met Ser Arg Gly Pro Glu Asp Val Ser Trp Pro Ser

9410 9415 9420

Pro Leu Ser Val Glu Lys Asn Ser Pro Ser Ser Leu Val Ser

9425 9430 9435

Ser Ser Ser Val Thr Ser Pro Ser Pro Leu Tyr Ser Thr Pro Ser

9440 9445 9450

Gly Ser Ser His Ser Ser Pro Val Pro Val Thr Ser Leu Phe Thr

9455 9460 9465

Ser Ile Met Met Lys Ala Thr Asp Met Leu Asp Ala Ser Leu Glu

9470 9475 9480

Pro Glu Thr Thr Ser Ala Pro Asn Met Asn Ile Thr Ser Asp Glu

9485 9490 9495

Ser Leu Ala Ala Ser Lys Ala Thr Thr Glu Thr Glu Ala Ile His

9500 9505 9510

Val Phe Glu Asn Thr Ala Ala Ser His Val Glu Thr Thr Ser Ala

9515 9520 9525

Thr Glu Glu Leu Tyr Ser Ser Ser Pro Gly Phe Ser Glu Pro Thr

9530 9535 9540

Lys Val Ile Ser Pro Val Val Thr Ser Ser Ser Ile Arg Asp Asn

9545 9550 9555

Met Val Ser Thr Thr Met Pro Gly Ser Ser Gly Ile Thr Arg Ile

9560 9565 9570

Glu Ile Glu Ser Met Ser Ser Leu Thr Pro Gly Leu Arg Glu Thr

9575 9580 9585

Arg Thr Ser Gln Asp Ile Thr Ser Ser Thr Glu Thr Ser Thr Val

9590 9595 9600

Leu Tyr Lys Met Pro Ser Gly Ala Thr Pro Glu Val Ser Arg Thr

9605 9610 9615

Glu Val Met Pro Ser Ser Arg Thr Ser Ile Pro Gly Pro Ala Gln

9620 9625 9630

Ser Thr Met Ser Leu Asp Ile Ser Asp Glu Val Val Thr Arg Leu

9635 9640 9645

Ser Thr Ser Pro Ile Met Thr Glu Ser Ala Glu Ile Thr Ile Thr

9650 9655 9660

Thr Gln Thr Gly Tyr Ser Leu Ala Thr Ser Gln Val Thr Leu Pro

9665 9670 9675

Leu Gly Thr Ser Met Thr Phe Leu Ser Gly Thr His Ser Thr Met

9680 9685 9690

Ser Gln Gly Leu Ser His Ser Glu Met Thr Asn Leu Met Ser Arg

9695 9700 9705

Gly Pro Glu Ser Leu Ser Trp Thr Ser Pro Arg Phe Val Glu Thr

9710 9715 9720

Thr Arg Ser Ser Ser Ser Leu Thr Ser Leu Pro Leu Thr Thr Ser

9725 9730 9735

Leu Ser Pro Val Ser Ser Thr Leu Leu Asp Ser Ser Pro Ser Ser

9740 9745 9750

Pro Leu Pro Val Thr Ser Leu Ile Leu Pro Gly Leu Val Lys Thr

9755 9760 9765

Thr Glu Val Leu Asp Thr Ser Ser Glu Pro Lys Thr Ser Ser Ser

9770 9775 9780

Pro Asn Leu Ser Ser Thr Ser Val Glu Ile Pro Ala Thr Ser Glu

9785 9790 9795

Ile Met Thr Asp Thr Glu Lys Ile His Pro Ser Ser Asn Thr Ala

9800 9805 9810

Val Ala Lys Val Arg Thr Ser Ser Ser Val His Glu Ser His Ser

9815 9820 9825

Ser Val Leu Ala Asp Ser Glu Thr Thr Ile Thr Ile Pro Ser Met

9830 9835 9840

Gly Ile Thr Ser Ala Val Asp Asp Thr Thr Val Phe Thr Ser Asn

9845 9850 9855

Pro Ala Phe Ser Glu Thr Arg Arg Ile Pro Thr Glu Pro Thr Phe

9860 9865 9870

Ser Leu Thr Pro Gly Phe Arg Glu Thr Ser Thr Ser Glu Glu Thr

9875 9880 9885

Thr Ser Ile Thr Glu Thr Ser Ala Val Leu Tyr Gly Val Pro Thr

9890 9895 9900

Ser Ala Thr Thr Glu Val Ser Met Thr Glu Ile Met Ser Ser Asn

9905 9910 9915

Arg Ile His Ile Pro Asp Ser Asp Gln Ser Thr Met Ser Pro Asp

9920 9925 9930

Ile Ile Thr Glu Val Ile Thr Arg Leu Ser Ser Ser Ser Met Met

9935 9940 9945

Ser Glu Ser Thr Gln Met Thr Ile Thr Thr Gln Lys Ser Ser Pro

9950 9955 9960

Gly Ala Thr Ala Gln Ser Thr Leu Thr Leu Ala Thr Thr Thr Ala

9965 9970 9975

Pro Leu Ala Arg Thr His Ser Thr Val Pro Arg Phe Leu His

9980 9985 9990

Ser Glu Met Thr Thr Leu Met Ser Arg Ser Pro Glu Asn Pro Ser

9995 10000 10005

Trp Lys Ser Ser Leu Phe Val Glu Lys Thr Ser Ser Ser Ser Ser

10010 10015 10020

Leu Leu Ser Leu Pro Val Thr Thr Ser Pro Ser Val Ser Ser Thr

10025 10030 10035

Leu Pro Gln Ser Ile Pro Ser Ser Ser Phe Ser Val Thr Ser Leu

10040 10045 10050

Leu Thr Pro Gly Met Val Lys Thr Thr Asp Thr Ser Thr Glu Pro

10055 10060 10065

Gly Thr Ser Leu Ser Pro Asn Leu Ser Gly Thr Ser Val Glu Ile

10070 10075 10080

Leu Ala Ala Ser Glu Val Thr Thr Asp Thr Glu Lys Ile His Pro

10085 10090 10095

Ser Ser Ser Met Ala Val Thr Asn Val Gly Thr Thr Ser Ser Gly

10100 10105 10110

His Glu Leu Tyr Ser Ser Val Ser Ile His Ser Glu Pro Ser Lys

10115 10120 10125

Ala Thr Tyr Pro Val Gly Thr Pro Ser Ser Met Ala Glu Thr Ser

10130 10135 10140

Ile Ser Thr Ser Met Pro Ala Asn Phe Glu Thr Thr Gly Phe Glu

10145 10150 10155

Ala Glu Pro Phe Ser His Leu Thr Ser Gly Phe Arg Lys Thr Asn

10160 10165 10170

Met Ser Leu Asp Thr Ser Ser Val Thr Pro Thr Asn Thr Pro Ser

10175 10180 10185

Ser Pro Gly Ser Thr His Leu Leu Gln Ser Ser Lys Thr Asp Phe

10190 10195 10200

Thr Ser Ser Ala Lys Thr Ser Ser Pro Asp Trp Pro Pro Ala Ser

10205 10210 10215

Gln Tyr Thr Glu Ile Pro Val Asp Ile Ile Thr Pro Phe Asn Ala

10220 10225 10230

Ser Pro Ser Ile Thr Glu Ser Thr Gly Ile Thr Ser Phe Pro Glu

10235 10240 10245

Ser Arg Phe Thr Met Ser Val Thr Glu Ser Thr His His Leu Ser

10250 10255 10260

Thr Asp Leu Leu Pro Ser Ala Glu Thr Ile Ser Thr Gly Thr Val

10265 10270 10275

Met Pro Ser Leu Ser Glu Ala Met Thr Ser Phe Ala Thr Thr Gly

10280 10285 10290

Val Pro Arg Ala Ile Ser Gly Ser Gly Ser Pro Phe Ser Arg Thr

10295 10300 10305

Glu Ser Gly Pro Gly Asp Ala Thr Leu Ser Thr Ile Ala Glu Ser

10310 10315 10320

Leu Pro Ser Ser Thr Pro Val Pro Phe Ser Ser Ser Thr Phe Thr

10325 10330 10335

Thr Thr Asp Ser Ser Thr Ile Pro Ala Leu His Glu Ile Thr Ser

10340 10345 10350

Ser Ser Ala Thr Pro Tyr Arg Val Asp Thr Ser Leu Gly Thr Glu

10355 10360 10365

Ser Ser Thr Thr Glu Gly Arg Leu Val Met Val Ser Thr Leu Asp

10370 10375 10380

Thr Ser Ser Gln Pro Gly Arg Thr Ser Ser Ser Pro Ile Leu Asp

10385 10390 10395

Thr Arg Met Thr Glu Ser Val Glu Leu Gly Thr Val Thr Ser Ala

10400 10405 10410

Tyr Gln Val Pro Ser Leu Ser Thr Arg Leu Thr Arg Thr Asp Gly

10415 10420 10425

Ile Met Glu His Ile Thr Lys Ile Pro Asn Glu Ala Ala His Arg

10430 10435 10440

Gly Thr Ile Arg Pro Val Lys Gly Pro Gln Thr Ser Thr Ser Pro

10445 10450 10455

Ala Ser Pro Lys Gly Leu His Thr Gly Gly Thr Lys Arg Met Glu

10460 10465 10470

Thr Thr Thr Thr Ala Leu Lys Thr Thr Thr Thr Thr Ala Leu Lys Thr

10475 10480 10485

Thr Ser Arg Ala Thr Leu Thr Thr Ser Val Tyr Thr Pro Thr Leu

10490 10495 10500

Gly Thr Leu Thr Pro Leu Asn Ala Ser Met Gln Met Ala Ser Thr

10505 10510 10515

Ile Pro Thr Glu Met Met Ile Thr Thr Pro Tyr Val Phe Pro Asp

10520 10525 10530

Val Pro Glu Thr Thr Ser Ser Leu Ala Thr Ser Leu Gly Ala Glu

10535 10540 10545

Thr Ser Thr Ala Leu Pro Arg Thr Thr Pro Ser Val Phe Asn Arg

10550 10555 10560

Glu Ser Glu Thr Thr Ala Ser Leu Val Ser Arg Ser Gly Ala Glu

10565 10570 10575

Arg Ser Pro Val Ile Gln Thr Leu Asp Val Ser Ser Ser Glu Pro

10580 10585 10590

Asp Thr Thr Ala Ser Trp Val Ile His Pro Ala Glu Thr Ile Pro

10595 10600 10605

Thr Val Ser Lys Thr Thr Pro Asn Phe Phe His Ser Glu Leu Asp

10610 10615 10620

Thr Val Ser Ser Thr Ala Thr Ser His Gly Ala Asp Val Ser Ser

10625 10630 10635

Ala Ile Pro Thr Asn Ile Ser Pro Ser Glu Leu Asp Ala Leu Thr

10640 10645 10650

Pro Leu Val Thr Ile Ser Gly Thr Asp Thr Ser Thr Thr Phe Pro

10655 10660 10665

Thr Leu Thr Lys Ser Pro His Glu Thr Glu Thr Arg Thr Thr Trp

10670 10675 10680

Leu Thr His Pro Ala Glu Thr Ser Ser Thr Ile Pro Arg Thr Ile

10685 10690 10695

Pro Asn Phe Ser His His Glu Ser Asp Ala Thr Pro Ser Ile Ala

10700 10705 10710

Thr Ser Pro Gly Ala Glu Thr Ser Ser Ala Ile Pro Ile Met Thr

10715 10720 10725

Val Ser Pro Gly Ala Glu Asp Leu Val Thr Ser Gln Val Thr Ser

10730 10735 10740

Ser Gly Thr Asp Arg Asn Met Thr Ile Pro Thr Leu Thr Leu Ser

10745 10750 10755

Pro Gly Glu Pro Lys Thr Ile Ala Ser Leu Val Thr His Pro Glu

10760 10765 10770

Ala Gln Thr Ser Ser Ala Ile Pro Thr Ser Thr Ile Ser Pro Ala

10775 10780 10785

Val Ser Arg Leu Val Thr Ser Met Val Thr Ser Leu Ala Ala Lys

10790 10795 10800

Thr Ser Thr Thr Asn Arg Ala Leu Thr Asn Ser Pro Gly Glu Pro

10805 10810 10815

Ala Thr Thr Val Ser Leu Val Thr His Pro Ala Gln Thr Ser Pro

10820 10825 10830

Thr Val Pro Trp Thr Thr Ser Ile Phe Phe His Ser Lys Ser Asp

10835 10840 10845

Thr Thr Pro Ser Met Thr Thr Ser His Gly Ala Glu Ser Ser Ser

10850 10855 10860

Ala Val Pro Thr Pro Thr Val Ser Thr Glu Val Pro Gly Val Val

10865 10870 10875

Thr Pro Leu Val Thr Ser Ser Arg Ala Val Ile Ser Thr Thr Ile

10880 10885 10890

Pro Ile Leu Thr Leu Ser Pro Gly Glu Pro Glu Thr Thr Pro Ser

10895 10900 10905

Met Ala Thr Ser His Gly Glu Glu Ala Ser Ser Ala Ile Pro Thr

10910 10915 10920

Pro Thr Val Ser Pro Gly Val Pro Gly Val Val Thr Ser Leu Val

10925 10930 10935

Thr Ser Ser Arg Ala Val Thr Ser Thr Thr Ile Pro Ile Leu Thr

10940 10945 10950

Phe Ser Leu Gly Glu Pro Glu Thr Thr Pro Ser Met Ala Thr Ser

10955 10960 10965

His Gly Thr Glu Ala Gly Ser Ala Val Pro Thr Val Leu Pro Glu

10970 10975 10980

Val Pro Gly Met Val Thr Ser Leu Val Ala Ser Ser Arg Ala Val

10985 10990 10995

Thr Ser Thr Thr Leu Pro Thr Leu Thr Leu Ser Pro Gly Glu Pro

11000 11005 11010

Glu Thr Thr Pro Ser Met Ala Thr Ser His Gly Ala Glu Ala Ser

11015 11020 11025

Ser Thr Val Pro Thr Val Ser Pro Glu Val Pro Gly Val Val Thr

11030 11035 11040

Ser Leu Val Thr Ser Ser Ser Gly Val Asn Ser Thr Ser Ile Pro

11045 11050 11055

Thr Leu Ile Leu Ser Pro Gly Glu Leu Glu Thr Thr Pro Ser Met

11060 11065 11070

Ala Thr Ser His Gly Ala Glu Ala Ser Ser Ala Val Pro Thr Pro

11075 11080 11085

Thr Val Ser Pro Gly Val Ser Gly Val Val Thr Pro Leu Val Thr

11090 11095 11100

Ser Ser Arg Ala Val Thr Ser Thr Thr Ile Pro Ile Leu Thr Leu

11105 11110 11115

Ser Ser Ser Glu Pro Glu Thr Thr Pro Ser Met Ala Thr Ser His

11120 11125 11130

Gly Val Glu Ala Ser Ser Ala Val Leu Thr Val Ser Pro Glu Val

11135 11140 11145

Pro Gly Met Val Thr Ser Leu Val Thr Ser Ser Arg Ala Val Thr

11150 11155 11160

Ser Thr Thr Ile Pro Thr Leu Thr Ile Ser Ser Asp Glu Pro Glu

11165 11170 11175

Thr Thr Thr Ser Leu Val Thr His Ser Glu Ala Lys Met Ile Ser

11180 11185 11190

Ala Ile Pro Thr Leu Ala Val Ser Pro Thr Val Gln Gly Leu Val

11195 11200 11205

Thr Ser Leu Val Thr Ser Ser Gly Ser Glu Thr Ser Ala Phe Ser

11210 11215 11220

Asn Leu Thr Val Ala Ser Ser Gln Pro Glu Thr Ile Asp Ser Trp

11225 11230 11235

Val Ala His Pro Gly Thr Glu Ala Ser Ser Val Val Pro Thr Leu

11240 11245 11250

Thr Val Ser Thr Gly Glu Pro Phe Thr Asn Ile Ser Leu Val Thr

11255 11260 11265

His Pro Ala Glu Ser Ser Ser Thr Leu Pro Arg Thr Thr Ser Arg

11270 11275 11280

Phe Ser His Ser Glu Leu Asp Thr Met Pro Ser Thr Val Thr Ser

11285 11290 11295

Pro Glu Ala Glu Ser Ser Ser Ala Ile Ser Thr Thr Ile Ser Pro

11300 11305 11310

Gly Ile Pro Gly Val Leu Thr Ser Leu Val Thr Ser Ser Gly Arg

11315 11320 11325

Asp Ile Ser Ala Thr Phe Pro Thr Val Pro Glu Ser Pro His Glu

11330 11335 11340

Ser Glu Ala Thr Ala Ser Trp Val Thr His Pro Ala Val Thr Ser

11345 11350 11355

Thr Thr Val Pro Arg Thr Thr Pro Asn Tyr Ser His Ser Glu Pro

11360 11365 11370

Asp Thr Thr Pro Ser Ile Ala Thr Ser Pro Gly Ala Glu Ala Thr

11375 11380 11385

Ser Asp Phe Pro Thr Ile Thr Val Ser Pro Asp Val Pro Asp Met

11390 11395 11400

Val Thr Ser Gln Val Thr Ser Ser Gly Thr Asp Thr Ser Ile Thr

11405 11410 11415

Ile Pro Thr Leu Thr Leu Ser Ser Gly Glu Pro Glu Thr Thr Thr

11420 11425 11430

Ser Phe Ile Thr Tyr Ser Glu Thr His Thr Ser Ser Ala Ile Pro

11435 11440 11445

Thr Leu Pro Val Ser Pro Gly Ala Ser Lys Met Leu Thr Ser Leu

11450 11455 11460

Val Ile Ser Ser Gly Thr Asp Ser Thr Thr Thr Phe Pro Thr Leu

11465 11470 11475

Thr Glu Thr Pro Tyr Glu Pro Glu Thr Thr Ala Ile Gln Leu Ile

11480 11485 11490

His Pro Ala Glu Thr Asn Thr Met Val Pro Arg Thr Thr Pro Lys

11495 11500 11505

Phe Ser His Ser Lys Ser Asp Thr Thr Leu Pro Val Ala Ile Thr

11510 11515 11520

Ser Pro Gly Pro Glu Ala Ser Ser Ala Val Ser Thr Thr Thr Ile

11525 11530 11535

Ser Pro Asp Met Ser Asp Leu Val Thr Ser Leu Val Pro Ser Ser

11540 11545 11550

Gly Thr Asp Thr Ser Thr Thr Phe Pro Thr Leu Ser Glu Thr Pro

11555 11560 11565

Tyr Glu Pro Glu Thr Thr Ala Thr Trp Leu Thr His Pro Ala Glu

11570 11575 11580

Thr Ser Thr Thr Val Ser Gly Thr Ile Pro Asn Phe Ser His Arg

11585 11590 11595

Gly Ser Asp Thr Ala Pro Ser Met Val Thr Ser Pro Gly Val Asp

11600 11605 11610

Thr Arg Ser Gly Val Pro Thr Thr Thr Ile Pro Pro Ser Ile Pro

11615 11620 11625

Gly Val Val Thr Ser Gln Val Thr Ser Ser Ala Thr Asp Thr Ser

11630 11635 11640

Thr Ala Ile Pro Thr Leu Thr Pro Ser Pro Gly Glu Pro Glu Thr

11645 11650 11655

Thr Ala Ser Ser Ala Thr His Pro Gly Thr Gln Thr Gly Phe Thr

11660 11665 11670

Val Pro Ile Arg Thr Val Pro Ser Ser Glu Pro Asp Thr Met Ala

11675 11680 11685

Ser Trp Val Thr His Pro Pro Gln Thr Ser Thr Pro Val Ser Arg

11690 11695 11700

Thr Thr Ser Ser Phe Ser His Ser Ser Pro Asp Ala Thr Pro Val

11705 11710 11715

Met Ala Thr Ser Pro Arg Thr Glu Ala Ser Ser Ala Val Leu Thr

11720 11725 11730

Thr Ile Ser Pro Gly Ala Pro Glu Met Val Thr Ser Gln Ile Thr

11735 11740 11745

Ser Ser Gly Ala Ala Thr Ser Thr Thr Val Pro Thr Leu Thr His

11750 11755 11760

Ser Pro Gly Met Pro Glu Thr Thr Ala Leu Leu Ser Thr His Pro

11765 11770 11775

Arg Thr Glu Thr Ser Lys Thr Phe Pro Ala Ser Thr Val Phe Pro

11780 11785 11790

Gln Val Ser Glu Thr Thr Ala Ser Leu Thr Ile Arg Pro Gly Ala

11795 11800 11805

Glu Thr Ser Thr Ala Leu Pro Thr Gln Thr Thr Ser Ser Leu Phe

11810 11815 11820

Thr Leu Leu Val Thr Gly Thr Ser Arg Val Asp Leu Ser Pro Thr

11825 11830 11835

Ala Ser Pro Gly Val Ser Ala Lys Thr Ala Pro Leu Ser Thr His

11840 11845 11850

Pro Gly Thr Glu Thr Ser Thr Met Ile Pro Thr Ser Thr Leu Ser

11855 11860 11865

Leu Gly Leu Leu Glu Thr Thr Gly Leu Leu Ala Thr Ser Ser Ser

11870 11875 11880

Ala Glu Thr Ser Thr Ser Thr Leu Thr Leu Thr Val Ser Pro Ala

11885 11890 11895

Val Ser Gly Leu Ser Ser Ala Ser Ile Thr Thr Asp Lys Pro Gln

11900 11905 11910

Thr Val Thr Ser Trp Asn Thr Glu Thr Ser Pro Ser Val Thr Ser

11915 11920 11925

Val Gly Pro Pro Glu Phe Ser Arg Thr Val Thr Gly Thr Thr Met

11930 11935 11940

Thr Leu Ile Pro Ser Glu Met Pro Thr Pro Pro Lys Thr Ser His

11945 11950 11955

Gly Glu Gly Val Ser Pro Thr Thr Ile Leu Arg Thr Thr Met Val

11960 11965 11970

Glu Ala Thr Asn Leu Ala Thr Thr Gly Ser Ser Pro Thr Val Ala

11975 11980 11985

Lys Thr Thr Thr Thr Thr Phe Asn Thr Leu Ala Gly Ser Leu Phe Thr

11990 11995 12000

Pro Leu Thr Thr Pro Gly Met Ser Thr Leu Ala Ser Glu Ser Val

12005 12010 12015

Thr Ser Arg Thr Ser Tyr Asn His Arg Ser Trp Ile Ser Thr Thr

12020 12025 12030

Ser Ser Tyr Asn Arg Arg Tyr Trp Thr Pro Ala Thr Ser Thr Pro

12035 12040 12045

Val Thr Ser Thr Phe Ser Pro Gly Ile Ser Thr Ser Ser Ile Pro

12050 12055 12060

Ser Ser Thr Ala Ala Thr Val Pro Phe Met Val Pro Phe Thr Leu

12065 12070 12075

Asn Phe Thr Ile Thr Asn Leu Gln Tyr Glu Glu Asp Met Arg His

12080 12085 12090

Pro Gly Ser Arg Lys Phe Asn Ala Thr Glu Arg Glu Leu Gln Gly

12095 12100 12105

Leu Leu Lys Pro Leu Phe Arg Asn Ser Ser Leu Glu Tyr Leu Tyr

12110 12115 12120

Ser Gly Cys Arg Leu Ala Ser Leu Arg Pro Glu Lys Asp Ser Ser

12125 12130 12135

Ala Thr Ala Val Asp Ala Ile Cys Thr His Arg Pro Asp Pro Glu

12140 12145 12150

Asp Leu Gly Leu Asp Arg Glu Arg Leu Tyr Trp Glu Leu Ser Asn

12155 12160 12165

Leu Thr Asn Gly Ile Gln Glu Leu Gly Pro Tyr Thr Leu Asp Arg

12170 12175 12180

Asn Ser Leu Tyr Val Asn Gly Phe Thr His Arg Ser Ser Met Pro

12185 12190 12195

Thr Thr Ser Thr Pro Gly Thr Ser Thr Val Asp Val Gly Thr Ser

12200 12205 12210

Gly Thr Pro Ser Ser Ser Pro Ser Pro Thr Thr Ala Gly Pro Leu

12215 12220 12225

Leu Met Pro Phe Thr Leu Asn Phe Thr Ile Thr Asn Leu Gln Tyr

12230 12235 12240

Glu Glu Asp Met Arg Arg Thr Gly Ser Arg Lys Phe Asn Thr Met

12245 12250 12255

Glu Ser Val Leu Gln Gly Leu Leu Lys Pro Leu Phe Lys Asn Thr

12260 12265 12270

Ser Val Gly Pro Leu Tyr Ser Gly Cys Arg Leu Thr Leu Leu Arg

12275 12280 12285

Pro Glu Lys Asp Gly Ala Ala Thr Gly Val Asp Ala Ile Cys Thr

12290 12295 12300

His Arg Leu Asp Pro Lys Ser Pro Gly Leu Asn Arg Glu Gln Leu

12305 12310 12315

Tyr Trp Glu Leu Ser Lys Leu Thr Asn Asp Ile Glu Glu Leu Gly

12320 12325 12330

Pro Tyr Thr Leu Asp Arg Asn Ser Leu Tyr Val Asn Gly Phe Thr

12335 12340 12345

His Gln Ser Ser Val Ser Thr Thr Ser Thr Pro Gly Thr Ser Thr

12350 12355 12360

Val Asp Leu Arg Thr Ser Gly Thr Pro Ser Ser Leu Ser Ser Pro

12365 12370 12375

Thr Ile Met Ala Ala Gly Pro Leu Leu Val Pro Phe Thr Leu Asn

12380 12385 12390

Phe Thr Ile Thr Asn Leu Gln Tyr Gly Glu Asp Met Gly His Pro

12395 12400 12405

Gly Ser Arg Lys Phe Asn Thr Thr Glu Arg Val Leu Gln Gly Leu

12410 12415 12420

Leu Gly Pro Ile Phe Lys Asn Thr Ser Val Gly Pro Leu Tyr Ser

12425 12430 12435

Gly Cys Arg Leu Thr Ser Leu Arg Ser Glu Lys Asp Gly Ala Ala

12440 12445 12450

Thr Gly Val Asp Ala Ile Cys Ile His His Leu Asp Pro Lys Ser

12455 12460 12465

Pro Gly Leu Asn Arg Glu Arg Leu Tyr Trp Glu Leu Ser Gln Leu

12470 12475 12480

Thr Asn Gly Ile Lys Glu Leu Gly Pro Tyr Thr Leu Asp Arg Asn

12485 12490 12495

Ser Leu Tyr Val Asn Gly Phe Thr His Arg Thr Ser Val Pro Thr

12500 12505 12510

Ser Ser Thr Pro Gly Thr Ser Thr Val Asp Leu Gly Thr Ser Gly

12515 12520 12525

Thr Pro Phe Ser Leu Pro Ser Pro Ala Thr Ala Gly Pro Leu Leu

12530 12535 12540

Val Leu Phe Thr Leu Asn Phe Thr Ile Thr Asn Leu Lys Tyr Glu

12545 12550 12555

Glu Asp Met His Arg Pro Gly Ser Arg Lys Phe Asn Thr Thr Glu

12560 12565 12570

Arg Val Leu Gln Thr Leu Leu Gly Pro Met Phe Lys Asn Thr Ser

12575 12580 12585

Val Gly Leu Leu Tyr Ser Gly Cys Arg Leu Thr Leu Leu Arg Ser

12590 12595 12600

Glu Lys Asp Gly Ala Ala Thr Gly Val Asp Ala Ile Cys Thr His

12605 12610 12615

Arg Leu Asp Pro Lys Ser Pro Gly Val Asp Arg Glu Gln Leu Tyr

12620 12625 12630

Trp Glu Leu Ser Gln Leu Thr Asn Gly Ile Lys Glu Leu Gly Pro

12635 12640 12645

Tyr Thr Leu Asp Arg Asn Ser Leu Tyr Val Asn Gly Phe Thr His

12650 12655 12660

Trp Ile Pro Val Pro Thr Ser Ser Thr Pro Gly Thr Ser Thr Val

12665 12670 12675

Asp Leu Gly Ser Gly Thr Pro Ser Ser Leu Pro Ser Pro Thr Thr

12680 12685 12690

Ala Gly Pro Leu Leu Val Pro Phe Thr Leu Asn Phe Thr Ile Thr

12695 12700 12705

Asn Leu Lys Tyr Glu Glu Asp Met His Cys Pro Gly Ser Arg Lys

12710 12715 12720

Phe Asn Thr Thr Glu Arg Val Leu Gln Ser Leu Leu Gly Pro Met

12725 12730 12735

Phe Lys Asn Thr Ser Val Gly Pro Leu Tyr Ser Gly Cys Arg Leu

12740 12745 12750

Thr Leu Leu Arg Ser Glu Lys Asp Gly Ala Ala Thr Gly Val Asp

12755 12760 12765

Ala Ile Cys Thr His Arg Leu Asp Pro Lys Ser Pro Gly Val Asp

12770 12775 12780

Arg Glu Gln Leu Tyr Trp Glu Leu Ser Gln Leu Thr Asn Gly Ile

12785 12790 12795

Lys Glu Leu Gly Pro Tyr Thr Leu Asp Arg Asn Ser Leu Tyr Val

12800 12805 12810

Asn Gly Phe Thr His Gln Thr Ser Ala Pro Asn Thr Ser Thr Pro

12815 12820 12825

Gly Thr Ser Thr Val Asp Leu Gly Thr Ser Gly Thr Pro Ser Ser

12830 12835 12840

Leu Pro Ser Pro Thr Ser Ala Gly Pro Leu Leu Val Pro Phe Thr

12845 12850 12855

Leu Asn Phe Thr Ile Thr Asn Leu Gln Tyr Glu Glu Asp Met His

12860 12865 12870

His Pro Gly Ser Arg Lys Phe Asn Thr Thr Glu Arg Val Leu Gln

12875 12880 12885

Gly Leu Leu Gly Pro Met Phe Lys Asn Thr Ser Val Gly Leu Leu

12890 12895 12900

Tyr Ser Gly Cys Arg Leu Thr Leu Leu Arg Pro Glu Lys Asn Gly

12905 12910 12915

Ala Ala Thr Gly Met Asp Ala Ile Cys Ser His Arg Leu Asp Pro

12920 12925 12930

Lys Ser Pro Gly Leu Asn Arg Glu Gln Leu Tyr Trp Glu Leu Ser

12935 12940 12945

Gln Leu Thr His Gly Ile Lys Glu Leu Gly Pro Tyr Thr Leu Asp

12950 12955 12960

Arg Asn Ser Leu Tyr Val Asn Gly Phe Thr His Arg Ser Ser Val

12965 12970 12975

Ala Pro Thr Ser Thr Pro Gly Thr Ser Thr Val Asp Leu Gly Thr

12980 12985 12990

Ser Gly Thr Pro Ser Ser Leu Pro Ser Pro Thr Thr Ala Val Pro

12995 13000 13005

Leu Leu Val Pro Phe Thr Leu Asn Phe Thr Ile Thr Asn Leu Gln

13010 13015 13020

Tyr Gly Glu Asp Met Arg His Pro Gly Ser Arg Lys Phe Asn Thr

13025 13030 13035

Thr Glu Arg Val Leu Gln Gly Leu Leu Gly Pro Leu Phe Lys Asn

13040 13045 13050

Ser Ser Val Gly Pro Leu Tyr Ser Gly Cys Arg Leu Ile Ser Leu

13055 13060 13065

Arg Ser Glu Lys Asp Gly Ala Ala Thr Gly Val Asp Ala Ile Cys

13070 13075 13080

Thr His His Leu Asn Pro Gln Ser Pro Gly Leu Asp Arg Glu Gln

13085 13090 13095

Leu Tyr Trp Gln Leu Ser Gln Met Thr Asn Gly Ile Lys Glu Leu

13100 13105 13110

Gly Pro Tyr Thr Leu Asp Arg Asn Ser Leu Tyr Val Asn Gly Phe

13115 13120 13125

Thr His Arg Ser Ser Gly Leu Thr Thr Ser Thr Pro Trp Thr Ser

13130 13135 13140

Thr Val Asp Leu Gly Thr Ser Gly Thr Pro Ser Pro Val Pro Ser

13145 13150 13155

Pro Thr Thr Thr Gly Pro Leu Leu Val Pro Phe Thr Leu Asn Phe

13160 13165 13170

Thr Ile Thr Asn Leu Gln Tyr Glu Glu Asn Met Gly His Pro Gly

13175 13180 13185

Ser Arg Lys Phe Asn Ile Thr Glu Ser Val Leu Gln Gly Leu Leu

13190 13195 13200

Lys Pro Leu Phe Lys Ser Thr Ser Val Gly Pro Leu Tyr Ser Gly

13205 13210 13215

Cys Arg Leu Thr Leu Leu Arg Pro Glu Lys Asp Gly Val Ala Thr

13220 13225 13230

Arg Val Asp Ala Ile Cys Thr His Arg Pro Asp Pro Lys Ile Pro

13235 13240 13245

Gly Leu Asp Arg Gln Gln Leu Tyr Trp Glu Leu Ser Gln Leu Thr

13250 13255 13260

His Ser Ile Thr Glu Leu Gly Pro Tyr Thr Leu Asp Arg Asp Ser

13265 13270 13275

Leu Tyr Val Asn Gly Phe Thr Gln Arg Ser Ser Val Pro Thr Thr

13280 13285 13290

Ser Thr Pro Gly Thr Phe Thr Val Gln Pro Glu Thr Ser Glu Thr

13295 13300 13305

Pro Ser Ser Leu Pro Gly Pro Thr Ala Thr Gly Pro Val Leu Leu

13310 13315 13320

Pro Phe Thr Leu Asn Phe Thr Ile Thr Asn Leu Gln Tyr Glu Glu

13325 13330 13335

Asp Met Arg Arg Pro Gly Ser Arg Lys Phe Asn Thr Thr Glu Arg

13340 13345 13350

Val Leu Gln Gly Leu Leu Met Pro Leu Phe Lys Asn Thr Ser Val

13355 13360 13365

Ser Ser Leu Tyr Ser Gly Cys Arg Leu Thr Leu Leu Arg Pro Glu

13370 13375 13380

Lys Asp Gly Ala Ala Thr Arg Val Asp Ala Val Cys Thr His Arg

13385 13390 13395

Pro Asp Pro Lys Ser Pro Gly Leu Asp Arg Glu Arg Leu Tyr Trp

13400 13405 13410

Lys Leu Ser Gln Leu Thr His Gly Ile Thr Glu Leu Gly Pro Tyr

13415 13420 13425

Thr Leu Asp Arg His Ser Leu Tyr Val Asn Gly Phe Thr His Gln

13430 13435 13440

Ser Ser Met Thr Thr Thr Arg Thr Pro Asp Thr Ser Thr Met His

13445 13450 13455

Leu Ala Thr Ser Arg Thr Pro Ala Ser Leu Ser Gly Pro Met Thr

13460 13465 13470

Ala Ser Pro Leu Leu Val Leu Phe Thr Ile Asn Phe Thr Ile Thr

13475 13480 13485

Asn Leu Arg Tyr Glu Glu Asn Met His His Pro Gly Ser Arg Lys

13490 13495 13500

Phe Asn Thr Thr Glu Arg Val Leu Gln Gly Leu Leu Arg Pro Val

13505 13510 13515

Phe Lys Asn Thr Ser Val Gly Pro Leu Tyr Ser Gly Cys Arg Leu

13520 13525 13530

Thr Leu Leu Arg Pro Lys Lys Asp Gly Ala Ala Thr Lys Val Asp

13535 13540 13545

Ala Ile Cys Thr Tyr Arg Pro Asp Pro Lys Ser Pro Gly Leu Asp

13550 13555 13560

Arg Glu Gln Leu Tyr Trp Glu Leu Ser Gln Leu Thr His Ser Ile

13565 13570 13575

Thr Glu Leu Gly Pro Tyr Thr Leu Asp Arg Asp Ser Leu Tyr Val

13580 13585 13590

Asn Gly Phe Thr Gln Arg Ser Ser Val Pro Thr Thr Ser Ile Pro

13595 13600 13605

Gly Thr Pro Thr Val Asp Leu Gly Thr Ser Gly Thr Pro Val Ser

13610 13615 13620

Lys Pro Gly Pro Ser Ala Ala Ser Pro Leu Leu Val Leu Phe Thr

13625 13630 13635

Leu Asn Phe Thr Ile Thr Asn Leu Arg Tyr Glu Glu Asn Met Gln

13640 13645 13650

His Pro Gly Ser Arg Lys Phe Asn Thr Thr Glu Arg Val Leu Gln

13655 13660 13665

Gly Leu Leu Arg Ser Leu Phe Lys Ser Thr Ser Val Gly Pro Leu

13670 13675 13680

Tyr Ser Gly Cys Arg Leu Thr Leu Leu Arg Pro Glu Lys Asp Gly

13685 13690 13695

Thr Ala Thr Gly Val Asp Ala Ile Cys Thr His His Pro Asp Pro

13700 13705 13710

Lys Ser Pro Arg Leu Asp Arg Glu Gln Leu Tyr Trp Glu Leu Ser

13715 13720 13725

Gln Leu Thr His Asn Ile Thr Glu Leu Gly Pro Tyr Ala Leu Asp

13730 13735 13740

Asn Asp Ser Leu Phe Val Asn Gly Phe Thr His Arg Ser Ser Val

13745 13750 13755

Ser Thr Thr Ser Thr Pro Gly Thr Pro Thr Val Tyr Leu Gly Ala

13760 13765 13770

Ser Lys Thr Pro Ala Ser Ile Phe Gly Pro Ser Ala Ala Ser His

13775 13780 13785

Leu Leu Ile Leu Phe Thr Leu Asn Phe Thr Ile Thr Asn Leu Arg

13790 13795 13800

Tyr Glu Glu Asn Met Trp Pro Gly Ser Arg Lys Phe Asn Thr Thr

13805 13810 13815

Glu Arg Val Leu Gln Gly Leu Leu Arg Pro Leu Phe Lys Asn Thr

13820 13825 13830

Ser Val Gly Pro Leu Tyr Ser Gly Cys Arg Leu Thr Leu Leu Arg

13835 13840 13845

Pro Glu Lys Asp Gly Glu Ala Thr Gly Val Asp Ala Ile Cys Thr

13850 13855 13860

His Arg Pro Asp Pro Thr Gly Pro Gly Leu Asp Arg Glu Gln Leu

13865 13870 13875

Tyr Leu Glu Leu Ser Gln Leu Thr His Ser Ile Thr Glu Leu Gly

13880 13885 13890

Pro Tyr Thr Leu Asp Arg Asp Ser Leu Tyr Val Asn Gly Phe Thr

13895 13900 13905

His Arg Ser Ser Val Pro Thr Thr Ser Thr Gly Val Val Ser Glu

13910 13915 13920

Glu Pro Phe Thr Leu Asn Phe Thr Ile Asn Asn Leu Arg Tyr Met

13925 13930 13935

Ala Asp Met Gly Gln Pro Gly Ser Leu Lys Phe Asn Ile Thr Asp

13940 13945 13950

Asn Val Met Gln His Leu Leu Ser Pro Leu Phe Gln Arg Ser Ser

13955 13960 13965

Leu Gly Ala Arg Tyr Thr Gly Cys Arg Val Ile Ala Leu Arg Ser

13970 13975 13980

Val Lys Asn Gly Ala Glu Thr Arg Val Asp Leu Leu Cys Thr Tyr

13985 13990 13995

Leu Gln Pro Leu Ser Gly Pro Gly Leu Pro Ile Lys Gln Val Phe

14000 14005 14010

His Glu Leu Ser Gln Gln Thr His Gly Ile Thr Arg Leu Gly Pro

14015 14020 14025

Tyr Ser Leu Asp Lys Asp Ser Leu Tyr Leu Asn Gly Tyr Asn Glu

14030 14035 14040

Pro Gly Pro Asp Glu Pro Pro Thr Thr Pro Lys Pro Ala Thr Thr

14045 14050 14055

Phe Leu Pro Pro Leu Ser Glu Ala Thr Thr Ala Met Gly Tyr His

14060 14065 14070

Leu Lys Thr Leu Thr Leu Asn Phe Thr Ile Ser Asn Leu Gln Tyr

14075 14080 14085

Ser Pro Asp Met Gly Lys Gly Ser Ala Thr Phe Asn Ser Thr Glu

14090 14095 14100

Gly Val Leu Gln His Leu Leu Arg Pro Leu Phe Gln Lys Ser Ser

14105 14110 14115

Met Gly Pro Phe Tyr Leu Gly Cys Gln Leu Ile Ser Leu Arg Pro

14120 14125 14130

Glu Lys Asp Gly Ala Ala Thr Gly Val Asp Thr Thr Cys Thr Tyr

14135 14140 14145

His Pro Asp Pro Val Gly Pro Gly Leu Asp Ile Gln Gln Leu Tyr

14150 14155 14160

Trp Glu Leu Ser Gln Leu Thr His Gly Val Thr Gln Leu Gly Phe

14165 14170 14175

Tyr Val Leu Asp Arg Asp Ser Leu Phe Ile Asn Gly Tyr Ala Pro

14180 14185 14190

Gln Asn Leu Ser Ile Arg Gly Glu Tyr Gln Ile Asn Phe His Ile

14195 14200 14205

Val Asn Trp Asn Leu Ser Asn Pro Asp Pro Thr Ser Ser Glu Tyr

14210 14215 14220

Ile Thr Leu Leu Arg Asp Ile Gln Asp Lys Val Thr Thr Leu Tyr

14225 14230 14235

Lys Gly Ser Gln Leu His Asp Thr Phe Arg Phe Cys Leu Val Thr

14240 14245 14250

Asn Leu Thr Met Asp Ser Val Leu Val Thr Val Lys Ala Leu Phe

14255 14260 14265

Ser Ser Asn Leu Asp Pro Ser Leu Val Glu Gln Val Phe Leu Asp

14270 14275 14280

Lys Thr Leu Asn Ala Ser Phe His Trp Leu Gly Ser Thr Tyr Gln

14285 14290 14295

Leu Val Asp Ile His Val Thr Glu Met Glu Ser Ser Val Tyr Gln

14300 14305 14310

Pro Thr Ser Ser Ser Ser Thr Gln His Phe Tyr Leu Asn Phe Thr

14315 14320 14325

Ile Thr Asn Leu Pro Tyr Ser Gln Asp Lys Ala Gln Pro Gly Thr

14330 14335 14340

Thr Asn Tyr Gln Arg Asn Lys Arg Asn Ile Glu Asp Ala Leu Asn

14345 14350 14355

Gln Leu Phe Arg Asn Ser Ser Ile Lys Ser Tyr Phe Ser Asp Cys

14360 14365 14370

Gln Val Ser Thr Phe Arg Ser Val Pro Asn Arg His His Thr Gly

14375 14380 14385

Val Asp Ser Leu Cys Asn Phe Ser Pro Leu Ala Arg Arg Val Asp

14390 14395 14400

Arg Val Ala Ile Tyr Glu Glu Phe Leu Arg Met Thr Arg Asn Gly

14405 14410 14415

Thr Gln Leu Gln Asn Phe Thr Leu Asp Arg Ser Ser Val Leu Val

14420 14425 14430

Asp Gly Tyr Ser Pro Asn Arg Asn Glu Pro Leu Thr Gly Asn Ser

14435 14440 14445

Asp Leu Pro Phe Trp Ala Val Ile Leu Ile Gly Leu Ala Gly Leu

14450 14455 14460

Leu Gly Val Ile Thr Cys Leu Ile Cys Gly Val Leu Val Thr Thr

14465 14470 14475

Arg Arg Arg Lys Lys Glu Gly Glu Tyr Asn Val Gln Gln Gln Cys

14480 14485 14490

Pro Gly Tyr Tyr Gln Ser His Leu Asp Leu Glu Asp Leu Gln

14495 14500 14505

<210> 9

<211> 490

<212> PRT

<213> Homo sapiens

<220>

<223> human protein STEAP2 (STAMP1)

<400> 9

Met Glu Ser Ile Ser Met Met Gly Ser Pro Lys Ser Leu Ser Glu Thr

1 5 10 15

Phe Leu Pro Asn Gly Ile Asn Gly Ile Lys Asp Ala Arg Lys Val Thr

20 25 30

Val Gly Val Ile Gly Ser Gly Asp Phe Ala Lys Ser Leu Thr Ile Arg

35 40 45

Leu Ile Arg Cys Gly Tyr His Val Val Ile Gly Ser Arg Asn Pro Lys

50 55 60

Phe Ala Ser Glu Phe Phe Pro His Val Val Asp Val Thr His His Glu

65 70 75 80

Asp Ala Leu Thr Lys Thr Asn Ile Ile Phe Val Ala Ile His Arg Glu

85 90 95

His Tyr Thr Ser Leu Trp Asp Leu Arg His Leu Leu Val Gly Lys Ile

100 105 110

Leu Ile Asp Val Ser Asn Asn Met Arg Ile Asn Gln Tyr Pro Glu Ser

115 120 125

Asn Ala Glu Tyr Leu Ala Ser Leu Phe Pro Asp Ser Leu Ile Val Lys

130 135 140

Gly Phe Asn Val Val Ser Ala Trp Ala Leu Gln Leu Gly Pro Lys Asp

145 150 155 160

Ala Ser Arg Gln Val Tyr Ile Cys Ser Asn Asn Ile Gln Ala Arg Gln

165 170 175

Gln Val Ile Glu Leu Ala Arg Gln Leu Asn Phe Ile Pro Ile Asp Leu

180 185 190

Gly Ser Leu Ser Ser Ala Arg Glu Ile Glu Asn Leu Pro Leu Arg Leu

195 200 205

Phe Thr Leu Trp Arg Gly Pro Val Val Val Ala Ile Ser Leu Ala Thr

210 215 220

Phe Phe Phe Leu Tyr Ser Phe Val Arg Asp Val Ile His Pro Tyr Ala

225 230 235 240

Arg Asn Gln Gln Ser Asp Phe Tyr Lys Ile Pro Ile Glu Ile Val Asn

245 250 255

Lys Thr Leu Pro Ile Val Ala Ile Thr Leu Leu Ser Leu Val Tyr Leu

260 265 270

Ala Gly Leu Leu Ala Ala Ala Tyr Gln Leu Tyr Tyr Gly Thr Lys Tyr

275 280 285

Arg Arg Phe Pro Pro Trp Leu Glu Thr Trp Leu Gln Cys Arg Lys Gln

290 295 300

Leu Gly Leu Leu Ser Phe Phe Phe Ala Met Val His Val Ala Tyr Ser

305 310 315 320

Leu Cys Leu Pro Met Arg Arg Ser Glu Arg Tyr Leu Phe Leu Asn Met

325 330 335

Ala Tyr Gln Gln Val His Ala Asn Ile Glu Asn Ser Trp Asn Glu Glu

340 345 350

Glu Val Trp Arg Ile Glu Met Tyr Ile Ser Phe Gly Ile Met Ser Leu

355 360 365

Gly Leu Leu Ser Leu Leu Ala Val Thr Ser Ile Pro Ser Val Ser Asn

370 375 380

Ala Leu Asn Trp Arg Glu Phe Ser Phe Ile Gln Ser Thr Leu Gly Tyr

385 390 395 400

Val Ala Leu Leu Ile Ser Thr Phe His Val Leu Ile Tyr Gly Trp Lys

405 410 415

Arg Ala Phe Glu Glu Glu Tyr Tyr Arg Phe Tyr Thr Pro Pro Asn Phe

420 425 430

Val Leu Ala Leu Val Leu Pro Ser Ile Val Ile Leu Gly Lys Ile Ile

435 440 445

Leu Phe Leu Pro Cys Ile Ser Arg Lys Leu Lys Arg Ile Lys Lys Gly

450 455 460

Trp Glu Lys Ser Gln Phe Leu Glu Glu Gly Met Gly Gly Thr Ile Pro

465 470 475 480

His Val Ser Pro Glu Arg Val Thr Val Met

485 490

<210> 10

<211> 453

<212> PRT

<213> Artificial sequence

<220>

<223> Heavy chain REGN1979 to CD20

<400> 10

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Asn Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Glu Trp Val

35 40 45

Ser Val Ile Ser Trp Asn Ser Asp Ser Ile Gly Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met His Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Lys Asp Asn His Tyr Gly Ser Gly Ser Tyr Tyr Tyr Tyr Gln Tyr

100 105 110

Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala

115 120 125

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser

130 135 140

Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

145 150 155 160

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

165 170 175

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

180 185 190

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr

195 200 205

Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg

210 215 220

Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Pro

225 230 235 240

Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

245 250 255

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

260 265 270

Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val

275 280 285

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser

290 295 300

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

305 310 315 320

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser

325 330 335

Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

340 345 350

Gln Val Tyr Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu

405 410 415

Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Leu Gly Lys

450

<210> 11

<211> 449

<212> PRT

<213> Artificial sequence

<220>

<223> Heavy chain REGN1979 to CD3

<400> 11

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Lys Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Lys Asp Asn Ser Gly Tyr Gly His Tyr Tyr Tyr Gly Met Asp Val

100 105 110

Trp Gly Gln Gly Thr Thr Val Thr Val Ala Ser Ala Ser Thr Lys Gly

115 120 125

Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser

130 135 140

Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val

145 150 155 160

Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe

165 170 175

Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val

180 185 190

Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val

195 200 205

Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys

210 215 220

Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn Arg Phe Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Lys

<210> 12

<211> 214

<212> PRT

<213> Artificial sequence

<220>

<223> Generic light chain REGN1979

<400> 12

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Tyr Ile Asn Trp Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 13

<211> 127

<212> PRT

<213> Artificial sequence

<220>

<223> HCVR REGN1979 to CD20

<400> 13

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Asn Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Glu Trp Val

35 40 45

Ser Val Ile Ser Trp Asn Ser Asp Ser Ile Gly Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met His Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Lys Asp Asn His Tyr Gly Ser Gly Ser Tyr Tyr Tyr Tyr Gln Tyr

100 105 110

Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120 125

<210> 14

<211> 123

<212> PRT

<213> Artificial sequence

<220>

<223> HCVR REGN1979 to CD3

<400> 14

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Lys Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Lys Asp Asn Ser Gly Tyr Gly His Tyr Tyr Tyr Gly Met Asp Val

100 105 110

Trp Gly Gln Gly Thr Thr Val Thr Val Ala Ser

115 120

<210> 15

<211> 108

<212> PRT

<213> Artificial sequence

<220>

<223> General LCVR REGN1979

<400> 15

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Tyr Ile Asn Trp Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg

100 105

<210> 16

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> HCDR1 REGN1979 to CD20

<400> 16

Gly Phe Thr Phe Asn Asp Tyr Ala

15

<210> 17

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> HCDR2 REGN1979 to CD20

<400> 17

Ile Ser Trp Asn Ser Asp Ser Ile

15

<210> 18

<211> 20

<212> PRT

<213> Artificial sequence

<220>

<223> HCDR3 REGN1979 to CD20

<400> 18

Ala Lys Asp Asn His Tyr Gly Ser Gly Ser Tyr Tyr Tyr Tyr Gln Tyr

1 5 10 15

Gly Met Asp Val

20

<210> 19

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> HCDR1 REGN1979 to CD3

<400> 19

Gly Phe Thr Phe Asp Asp Tyr Thr

15

<210> 20

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> HCDR2 REGN1979 to CD3

<400> 20

Ile Ser Trp Asn Ser Gly Ser Ile

15

<210> 21

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> HCDR3 REGN1979 to CD3

<400> 21

Ala Lys Asp Asn Ser Gly Tyr Gly His Tyr Tyr Tyr Gly Met Asp Val

1 5 10 15

<210> 22

<211> 6

<212> PRT

<213> Artificial sequence

<220>

<223> Common LCDR1 REGN1979

<400> 22

Gln Ser Val Ser Ser Asn

15

<210> 23

<211> 3

<212> PRT

<213> Artificial sequence

<220>

<223> Common LCDR2 REGN1979

<400> 23

Gly Ala Ser

1

<210> 24

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Common LCDR3 REGN1979

<400> 24

Gln His Tyr Ile Asn Trp Pro Leu Thr

15

<---

Claims (94)

1. A method of administering to a subject a therapeutic protein according to a dosing schedule to reduce the incidence or severity of a cytokine release syndrome or infusion reaction in the treatment of CD20-expressing B-cell lymphoma, comprising: administration of primary dose portions (D1) of said therapeutic protein at week 1 of the dosing regimen, where said primary dose is not more than 10 mg of said therapeutic protein, the first dose portion (F1D1) is between 40% and 60% of the total primary dose and is administered to the subject at day 1 of week 1, and the second portion of the dose (F2D1) is the remaining 40% to 60% of the total primary dose, and is administered to the subject 12-96 hours after administration of F1D1; administration of portions of a secondary dose (D2) of said therapeutic protein at week 2 of the dosing regimen, where said secondary dose is higher than the primary dose and is no more than half of the maximum weekly dose of said therapeutic protein, the first portion of the dose (F1D2) is 40% up to 60% of the total secondary dose, the second portion of the dose (F2D2) is the remaining 40% to 60% of the total secondary dose, and F2D2 is administered to the subject 12-96 hours after F1D2 administration in week 2 of the dosing regimen; And administering to the subject the specified maximum weekly dose of the specified therapeutic protein in the form of a single dose on one of the subsequent weeks of the dosing regimen. 2. The method of claim 1 wherein F2D1 is administered to the subject 24-96 hours after F1D1 is administered. 3. The method of claim 1 wherein F2D1 is administered to the subject 18-72 hours after F1D1 is administered. 4. The method according to any one of paragraphs. 1-3, where F2D2 is administered to a subject 24-96 hours after administration of F1D2. 5. The method according to any one of paragraphs. 1-3, where F2D2 is administered to a subject 18-72 hours after administration of F1D2. 6. The method according to any one of paragraphs. 1-5, where the indicated subsequent week is week 3 of the dosing regimen. 7. The method according to any one of paragraphs. 1-5, where the indicated subsequent week is week 4 of the dosing regimen. 8. The method according to any one of paragraphs. 1-5, where the indicated subsequent week is week 14 of the dosing regimen. 9. The method according to any one of paragraphs. 1-5, where said subsequent week is any of weeks 4-36 of the dosing regimen. 10. The method according to any one of paragraphs. 1-5, further comprising: administration of portions of a tertiary dose (D3) of said therapeutic protein at week 1 of the dosing regimen, where said tertiary dose is at least half of the specified maximum weekly dose of said therapeutic protein and not more than the specified maximum weekly dose of said therapeutic protein, the first portion of the dose (F1D3) is from 40% to 60% of the total tertiary dose, the second portion of the dose (F2D3) is the remaining 40% to 60% of the total tertiary dose, and F2D3 is administered to the subject 12-96 hours after F1D3 administration in week 1 of the dosing regimen; And administering to the subject the specified maximum weekly dose of the specified therapeutic protein in the form of a single dose on one of the subsequent weeks of the dosing regimen. 11. The method of claim 10, wherein the F2D3 is administered to the subject 24-96 hours after the administration of the F1D3. 12. The method of claim 10, wherein the F2D3 is administered to the subject 18-72 hours after the administration of the F1D3. 13. The method according to any one of paragraphs. 10-12, where the subsequent week indicated is week 4 of the dosing regimen. 14. The method according to any one of paragraphs. 10-12, where the subsequent week indicated is week 14 of the dosing regimen. 15. The method according to any one of paragraphs. 10-12, where said subsequent week is any of weeks 4-36 of the dosing regimen. 16. The method according to any one of paragraphs. 10-12 wherein the tertiary dose is administered as a single dose at weeks 4-12 of the dosing regimen. 17. A method of administering to a subject a therapeutic protein according to a dosing schedule to reduce the incidence or severity of a cytokine release syndrome or infusion reaction in the treatment of CD20-expressing B-cell lymphoma, comprising: administration of primary dose portions (D1) of said therapeutic protein at week 1 of the dosing regimen, where said primary dose is not more than 10 mg of said therapeutic protein, the first dose portion (F1D1) is between 40% and 60% of the total primary dose and is administered to the subject at day 1 of week 1, and the second portion of the dose (F2D1) is the remaining 40% to 60% of the total primary dose and is administered to the subject 12-96 hours after administration of F1D1; administration of portions of a secondary dose (D2) of said therapeutic protein at week 2 of the dosing regimen, where said secondary dose is equal to the maximum weekly dose of said therapeutic protein, the first portion of the dose (F1D2) is 40% to 60% of the total secondary dose, the second portion of the dose (F2D2 ) represents the remaining 40% to 60% of the total secondary dose, and F2D2 is administered to the subject 12-96 hours after F1D2 administration at week 2 of the dosing regimen; And administering to the subject the specified maximum weekly dose of the specified therapeutic protein in the form of a single dose on one of the subsequent weeks of the dosing regimen. 18. The method according to any one of paragraphs. 1-17 wherein said 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 the weekly phase of the dosing regimen. 19. The method according to any one of paragraphs. 1-18 further comprising administering to the subject the specified maximum weekly dose of the therapeutic protein as a biweekly single dose during the maintenance phase of the dosing regimen following the completion of the weekly phase of the dosing regimen. 20. The method according to any one of paragraphs. 1-18 further comprising administering to the subject the specified maximum weekly dose of the therapeutic protein as a single dose every three weeks during the maintenance phase of the dosing regimen following the completion of the weekly phase of the dosing regimen. 21. The method according to any one of paragraphs. 1-18 further comprising administering to the subject the specified maximum weekly dose of the therapeutic protein as a single dose every four weeks during the maintenance phase of the dosing regimen following the completion of the weekly phase of the dosing regimen. 22. The method according to any one of paragraphs. 19-21, where the maintenance phase is at least 24 weeks. 23. The method of claim 22 wherein the maintenance phase is 24 weeks. 24. The method according to any one of paragraphs. 1-23, where the indicated primary dose (D1) is 1 mg. 25. The method according to any one of paragraphs. 1-24, where the indicated secondary dose (D2) is 20 mg. 26. The method according to any one of paragraphs. 10-16 or 18-25, where the specified tertiary dose is 40 mg. 27. The method according to any one of paragraphs. 10-16 or 18-25, where the specified tertiary dose is 80 mg. 28. The method according to any one of paragraphs. 10-16 or 18-25, where the specified tertiary dose is 160 mg. 29. The method according to any one of paragraphs. 10-16 or 18-25, where the indicated tertiary dose is 320 mg. 30. The method according to any one of paragraphs. 1-29, where F1D1 is 50% of the total primary dose and F2D1 is 50% of the total primary dose. 31. The method according to any one of paragraphs. 1-30, where F1D2 is 50% of the total secondary dose and F2D2 is 50% of the total secondary dose. 32. The method according to any one of paragraphs. 10-16 or 18-31, where F1D3 is 50% of the total tertiary dose and F2D3 is 50% of the total tertiary dose. 33. The method according to any one of paragraphs. 1-32, where the specified maximum weekly dose of therapeutic protein is from 5 to 320 mg. 34. The method according to p. 33, where the specified maximum weekly dose of 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 said maximum weekly dose is 80 mg. 36. The method of claim 34 wherein said maximum weekly dose is 160 mg. 37. The method of claim 34, wherein said maximum weekly dose is 320 mg. 38. The method according to any one of paragraphs. 1-37, wherein each dose or portion of a dose is administered to a subject over a period of 1 to 6 hours. 39. The method according to any one of paragraphs. 1-38, wherein the CD20-expressing B cell lymphoma is non-Hodgkin's lymphoma, diffuse large B cell lymphoma, follicular lymphoma, mantle cell or marginal zone lymphoma, Burkitt's lymphoma, primary mediastinal B cell lymphoma. 40. The method according to any one of paragraphs. 1-38, wherein the subject is diagnosed with follicular lymphoma (FL). 41. The method according to p. 40, where the PL has a degree of 1-3a. 42. The method according to any one of paragraphs. 1-38, wherein the subject is diagnosed with diffuse large B-cell lymphoma (DLBCL). 43. The method of claim 42, wherein the subject has failed prior CAR-T therapy. 44. The method according to any one of paragraphs. 1-38, wherein the subject is diagnosed with mantle cell lymphoma (MCL). 45. The method of claim 44, wherein the subject has failed prior therapy with a Bruton's tyrosine kinase (BTK) inhibitor. 46. The method according to any one of paragraphs. 1-38, wherein the subject is diagnosed with marginal zone lymphoma (LMZ). 47. The method according to any one of paragraphs. 1-46, where the subject is an adult. 48. The method according to any one of paragraphs. 1-47, where the therapeutic protein is an antibody or antigen-binding fragment. 49. The method of claim 48, wherein the antibody is a fully human antibody. 50. The method of claim 48 or 49, wherein the antibody is a bispecific antibody or an antigen-binding fragment thereof. 51. The method of claim 50, wherein the first antigen-binding arm of the bispecific antibody or antigen-binding fragment binds to a T-cell antigen. 52. The method of claim 51 wherein the T cell antigen is CD3. 53. The method of claim 51 wherein the T cell antigen is CD28. 54. The method according to any one of paragraphs. 50-53, wherein the second antigen-binding arm of the bispecific antibody or antigen-binding fragment binds to CD20. 55. The method of claim 54 wherein the bispecific antibody is an anti-CD20 x anti-CD3 antibody. 56. The method of claim 55 wherein the anti-CD20 x anti-CD3 antibody is a REGN1979 antibody or an antibody containing HCVR/LCVR REGN1979 regions or an antibody containing REGN1979 CDR regions. 57. The method according to any one of paragraphs. 1-56, wherein the serum concentration of the therapeutic protein is maintained at or above about 2000 μg/L after the indicated maximum weekly dose is administered throughout the dosing regimen. 58. The method of claim 57, wherein the serum concentration of the therapeutic protein is maintained at or above about 2600 μg/l after the indicated maximum weekly dose is administered throughout the dosing regimen. 59. The method of claim 58, wherein the mean serum concentration of the therapeutic protein is maintained at at least about 3700 μg/L following the administration of said maximum weekly dose throughout the dosing regimen. 60. The method according to any one of paragraphs. 1-59, wherein 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 (NSAID), an IL-6 antagonist, or an IL-6R antagonist. 61. The method of claim 60 wherein the steroid is dexamethasone or the NSAID is indomethacin. 62. The method of claim 61, wherein the dexamethasone is administered to the subject approximately one to three hours prior to the first portion of the dose (F1D1). 63. The method of claim 61, wherein the dexamethasone is administered to the subject approximately one to three hours prior to F1D1, F2D1, F1D2, and F2D2. 64. The method of claim 61, wherein the dexamethasone is administered to the subject approximately one to three hours prior to F1D1, F2D1, F1D2, F2D2, F1D3, and F2D3. 65. The method of claim 60 wherein the IL-6 antagonist is an anti-IL-6 antibody or the IL-6R antagonist is an anti-IL-6R antibody. 66. The method of claim 65 wherein the anti-IL-6R antibody is sarilumab. 67. The method according to any one of paragraphs. 60-66, where the administration of the second agent is stopped after the first administration of the specified maximum weekly dose throughout the dosing regimen. 68. The method according to any one of paragraphs. 1-67, wherein the therapeutic protein is administered to a subject in combination with a second therapeutic agent. 69. The method of claim 68 wherein the second therapeutic agent comprises at least one of rituximab, obinutuzumab, cyclophosphamide, doxorubicin, vincristine, prednisone, prednisolone, bendamustine, lenalidomide, chlorambucil, ibritumomab tiuxetan, idelicib, copanlisib, duvelisib, etoposide, methyl prednisolone , cytarabine, cisplatin, mesna, ifosfamide, mitoxantrone, and procarbazine. 70. The method of claim 68 wherein the second therapeutic agent comprises a combination of cyclophosphamide, doxorubicin, vincristine, and prednisone. 71. The method of claim 68 wherein the second therapeutic agent comprises a combination of ifosfamide, cisplatin and etoposide. 72. The method of claim 68 wherein the second therapeutic agent comprises a combination of gemcitabine and oxaliplatin. 73. The method of claim 68 wherein the second therapeutic agent comprises a combination of lenalidomide and rituximab. 74. The method of claim 68 wherein the second therapeutic agent is lenalidomide. 75. A method of treating CD20-expressing B-cell lymphoma in a subject, comprising: selecting a subject diagnosed with CD20-expressing B-cell lymphoma; And administering to said subject a therapeutic protein in accordance with the method of any one of paragraphs. 1-74. 76. The method of claim 75, wherein the subject has previously received anti-cancer therapy. 77. The method of claim 75 or 76 wherein the subject is refractory to prior treatment or has relapsed after prior treatment. 78. The method of claim 75, wherein the subject has previously received treatment with an anti-CD20 antibody. 79. The method according to any one of paragraphs. 75-78, where the subject has previously received therapy using CAR-T. 80. The method according to any one of paragraphs. 75-79, 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. 81. The method according to any one of paragraphs. 1-80, where the incidence of CRS grade 3 and IR is less than 10%. 82. The method according to claim 81, where the incidence of CRS grade 3 and IR is less than 7.5% or less than 7%. 83. The method according to p. 81 or 82, where the specified maximum weekly dose is 80 mg or more. 84. The method according to any one of paragraphs. 1-83, wherein any dose administered as a single dose is administered no more than 1 hour apart.

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