TW202021625A - Methods of using anti-cd79b immunoconjugates - Google Patents

Abstract

Provided herein are methods of treating B-cell proliferative disorders (such as Diffuse Large B-Cell Lymphoma “DLBCL”) using immunoconjugates comprising anti-CD79b antibodies in combination with an alkylating agent (such as bendamustine) and an anti-CD20 antibody (such as rituximab).

Description

Method of using anti-CD79B immunoconjugate

The present invention relates to the treatment of B cell proliferative diseases by administering immunoconjugates containing anti-CD79b antibodies and alkylating agents (such as bendamustine) and anti-CD20 antibodies (such as rituximab), such as Diffuse large B-cell lymphoma (DLBCL) method.

Diffuse large B-cell lymphoma (DLBCL) accounts for approximately 25% of all newly diagnosed non-Hodgkin lymphoma cases (Armitage et al., Journal of Clinical Oncology, 16:2780-95, 1998; Swerdlow et al., International Agency for Research on Cancer (IARC), revised fourth edition, 2017). For treatment with rituximab, cyclophosphamide, adriamycin, vincristine and prednisone (R-CHOP) chemotherapy as the current standard of care, 30-40% of patients are refractory Or relapse after this treatment (Vitolo et al., Journal of Clinical Oncology, 25:3529-37, 2017; Coiffier et al., New England Journal of Medicine, 346:235-42, 2002). A higher treatment failure rate was observed in the lower-risk subgroups, including activated B-cell-like (ABC) and MYC/BCL2 dual manifestation lymphoma (DEL) (Scott et al., Journal of Clinical Oncology, 33: 2848- 56, 2015; Johnson et al., Journal of Clinical Oncology, 30:3452-59, 2012). For relapsed/refractory (R/R) patients, platinum rescue therapy followed by high-dose chemotherapy and autologous stem cell transplantation (ASCT) can cure up to 30-40% of patients who can withstand this therapy (Gisselbrecht et al., Journal of Clinical Oncology, 28:4184-90, 2010; Crump et al., Blood, 130:1800-08, 2017). However, for most R/R DLBCL patients who are not suitable for ASCT due to age, comorbidities, or insufficient response to salvage chemotherapy, and for relapse after ASCT, the median overall survival (OS) is approximately 6 months Patients have a poor prognosis (Czuczman et al., Clinical Cancer Research, 23:4127-37, 2017). There is no standard of care in this situation.

Recently, in the United States and the European Union, CD19-guided chimeric antigen receptor (CAR)-T cell therapy has been approved for use in third-line or later settings (Neelapu et al., New England Journal of Medicine, 377:2531-44, 2017 ; Schuster et al., Blood, 130:577, 2017). Although CAR-T cell therapy looks promising, its widespread use is restricted due to the lack of effective bridging therapy, therapeutic toxicity, and limited access due to high costs and the demand for specialized centers. Therefore, there are still significant unmet medical needs for R/R DLBCL patients who are not suitable for transplantation, including those patients who have failed ASCT. The present invention addresses this need and other needs.

Provided herein are methods and uses of anti-CD79b immunoconjugates for the treatment of B cell proliferative diseases in individuals in need thereof (eg, human individuals). Specifically, these methods and uses are based on polatuxumab vedotin and alkylating agents (such as bendamustine, such as bendamustine-HCl) and anti-CD20 antibodies (such as rituximab). Data from a randomized phase II clinical study in DLBCL patients who have previously received at least one DLBCL treatment. This study shows that it is comparable to treatment with alkylating agents (such as bendamustine, such as bendamustine-HCl) and anti-CD20 antibodies (such as rituximab) in the absence of anti-CD79b immunoconjugates Compared with a combination of an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin), an alkylating agent (e.g., bendamustine, e.g., bendamustine-HCl), and an anti-CD20 antibody (e.g., rituximab) Treatment reduces the risk of disease progression or death (PFS). In addition, compared with patients who received only alkylating agents (such as bendamustine, such as bendamustine-HCl) and anti-CD20 antibodies (such as rituximab), patients who received anti-CD79b immunoconjugates (such as , Polatuzumab vedotin), alkylating agents (for example, bendamustine, such as bendamustine-HCl) and anti-CD20 antibodies (for example, rituximab) patients showed statistically significant overall survival The improvement. The safety of anti-CD79b immunoconjugates (e.g., polatuzumab vedotin), alkylating agents (e.g. bendamustine, e.g. bendamustine-HCl) and anti-CD20 antibodies (e.g. rituximab) in combination seems to be the same The known safety characteristics of individual drugs are consistent, and no new safety signals have been identified for the combination.

, 其中Ab係抗CD79b抗體，其包含(i)包含SEQ ID NO：21之胺基酸序列之HVR-H1；(ii)包含SEQ ID NO：22之胺基酸序列之HVR-H2；(iii)包含SEQ ID NO：23之胺基酸序列之HVR-H3；(iv)包含SEQ ID NO：24之胺基酸序列之HVR-L1；(v)包含SEQ ID NO：25之胺基酸序列之HVR-L2；(vi)包含SEQ ID NO：26之胺基酸序列之HVR-L3，並且其中p為1至8，b)烷化劑，及(c)抗CD20抗體，其中該治療延長人之無進展生存期(PFS)。在一些實施例中，治療延長了個體之總生存期(OS)。This article provides a method for the treatment of diffuse large B-cell lymphoma (DLBCL) in a person in need, including administering an effective amount of: (a) an immunoconjugate of the following formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iii) ) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25 HVR-L2; (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and wherein p is 1 to 8, b) alkylating agent, and (c) anti-CD20 antibody, wherein the treatment is prolonged Human progression-free survival (PFS). In some embodiments, treatment prolongs the overall survival (OS) of the individual.

其中Ab係抗CD79b抗體，其包含(i)包含SEQ ID NO：21之胺基酸序列之HVR-H1；(ii)包含SEQ ID NO：22之胺基酸序列之HVR-H2；(iii)包含SEQ ID NO：23之胺基酸序列之HVR-H3；(iv)包含SEQ ID NO：24之胺基酸序列之HVR-L1；(v)包含SEQ ID NO：25之胺基酸序列之HVR-L2；(vi)包含SEQ ID NO：26之胺基酸序列之HVR-L3，並且其中p為1至8，(b)烷化劑，及(c)抗CD20抗體，其中該治療延長人之總生存期(OS)。The present invention provides a method for treating diffuse large B-cell lymphoma (DLBCL) in a person in need, comprising administering an effective amount of: (a) an immunoconjugate of the following formula

Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25 HVR-L2; (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and wherein p is 1 to 8, (b) alkylating agent, and (c) anti-CD20 antibody, wherein the treatment is prolonged Human overall survival (OS).

In some embodiments, humans reach a complete response (CR) after treatment with immunoconjugates, alkylating agents, and anti-CD20 antibodies. In some embodiments, the anti-CD79 antibody comprises: (i) VH, which comprises the amino acid sequence of SEQ ID NO: 19; and (ii) VL, which comprises the amino acid sequence of SEQ ID NO: 20. In some embodiments, the anti-CD79 antibody comprises (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 36 and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 35. In some embodiments, the immunoconjugate is polatuzumab vedotin. In some embodiments, the alkylating agent is 4-[5-[bis(2-chloroethyl)amino]-1-methylbenzimidazol-2-yl]butyric acid or a salt thereof. In some embodiments, the alkylating agent is bendamustine or a salt thereof. In some embodiments, the alkylating agent is bendamustine-HCl. In some embodiments, the anti-CD20 antibody system is rituximab, humanized B-Ly1 antibody (eg, atolizumab), ofatumumab, ublituximab, and/or iblituximab.

In some embodiments, the immunoconjugate is administered at a dosage of 1.8 mg/kg, the alkylating agent is administered at a dosage of 90 mg/m ² , and the anti-CD20 antibody is administered at a dosage of 375 mg/m ² . In some embodiments, the immunoconjugate, alkylating agent, and anti-CD20 antibody are administered for at least 6 21-day cycles, wherein during the 21-day cycle of the first cycle, the immunoconjugate is administered at a dose of 1.8 mg/kg on the second day For intravenous administration, the alkylating agent was administered intravenously at a dose of 90 mg/m ² on the second and third days, and the anti-CD20 antibody was administered intravenously at a dose of 375 mg/m ² on the first day. And in each 21-day cycle of cycles 2-6, the immunoconjugate is administered intravenously at a dose of 1.8 mg/kg on the first day, and the alkylating agent is administered at 90 mg/m ² on the first and second days The anti-CD20 antibody was administered intravenously at a dose of 375 mg/m ² on the first day. In some embodiments, the immunoconjugate and the alkylating agent are administered sequentially on the second day of the first cycle. In some embodiments, the immunoconjugate is administered before the alkylating agent. In some embodiments, the immunoconjugate, alkylating agent, and anti-CD20 antibody are administered sequentially on day 1 of cycles 2-6. In some embodiments, the anti-CD20 antibody is administered before the immunoconjugate, and wherein the immunoconjugate is administered before the alkylating agent on Day 1 of Cycles 2-6. In some embodiments, the immunoconjugate, alkylating agent and anti-CD20 antibody are further administered after the sixth cycle. In some embodiments, in each cycle after the 6th cycle, in each 21-day cycle, the immunoconjugate is administered intravenously at a dose of 1.8 mg/kg on day 1, and the alkylating agent is administered on day 1 and The anti-CD20 antibody was administered intravenously at a dose of 90 mg/m ^{2 on} the second day, and the anti-CD20 antibody was administered intravenously at a dose of 375 mg/m ² on the first day. In some embodiments, the anti-CD20 antibody is administered before the immunoconjugate, and wherein the immunoconjugate is administered before the alkylating agent on day 1 of each 21-day cycle of each cycle after the sixth cycle. Other exemplary dosing and administration regimens are provided elsewhere herein.

, 其中Ab係抗CD79b抗體，其包含(i)包含VH之重鏈，該VH包含SEQ ID NO：19之胺基酸序列，及(ii)包含VL之輕鏈，該VL包含SEQ ID NO：20之胺基酸序列，其中p為2至5，(b)苯達莫司汀或其鹽，及(c)利妥昔單抗，其中免疫結合物以1.8 mg/kg之劑量投與，苯達莫司汀或其鹽以90 mg/m² 之劑量投與，利妥昔單抗以375 mg/m² 之劑量投與，其中該治療延長了人之無進展生存期(PFS)及/或總生存期(OS)，並且其中：i) DLBCL係活化之B細胞樣DLBCL (ABC-DLBCL)或生髮中心B細胞樣DLBCL (GCB-BLBCL); iii) DLBCL係雙表現淋巴瘤(DEL); iv)人至少接受過兩種DLBCL先前療法; v)人至少接受過三種DLBCL先前療法;及/或vi)人已經接受了超過三種DLBCL先前療法。在一些實施例中，p介於3與4之間(例如，3.5)。在一些實施例中，重鏈包含胺基酸序列SEQ ID NO: 36，並且其中輕鏈包含胺基酸序列SEQ ID NO: 35。在一些實施例中，苯達莫司汀或其鹽係苯達莫司汀-HCl。在一些實施例中，在用免疫結合物、苯達莫司汀或其鹽及利妥昔單抗治療後，人達成完全響應(CR)。The present invention provides a method for treating diffuse large B-cell lymphoma in a person in need, comprising administering an effective amount of: (a) an immunoconjugate of the following formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) a heavy chain comprising VH, which VH comprises the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, which VL comprises SEQ ID NO: The amino acid sequence of 20, where p is 2 to 5, (b) bendamustine or its salt, and (c) rituximab, wherein the immunoconjugate is administered at a dose of 1.8 mg/kg, Bendamustine or its salt is administered at a dose of 90 mg/m ² , and rituximab is administered at a dose of 375 mg/m ^2. The treatment prolongs the progression-free survival (PFS) and / Or overall survival (OS), and in which: i) DLBCL line of activated B-cell-like DLBCL (ABC-DLBCL) or germinal center B-cell-like DLBCL (GCB-BLBCL); iii) DLBCL line of dual manifestation lymphoma (DEL ); iv) the person has received at least two prior therapies for DLBCL; v) the person has received at least three prior therapies for DLBCL; and/or vi) the person has received more than three prior therapies for DLBCL. In some embodiments, p is between 3 and 4 (eg, 3.5). In some embodiments, the heavy chain comprises the amino acid sequence of SEQ ID NO: 36, and wherein the light chain comprises the amino acid sequence of SEQ ID NO: 35. In some embodiments, bendamustine or its salt is bendamustine-HCl. In some embodiments, the person achieves a complete response (CR) after treatment with an immunoconjugate, bendamustine or its salt, and rituximab.

In some embodiments, the immunoconjugate, bendamustine or a salt thereof, and rituximab are administered for at least 6 21-day cycles, wherein the immunoconjugate is administered intravenously at a dose of 1.8 mg/kg on day 2. Administration, bendamustine or its salt was administered intravenously at a dose of 90 mg/m ² on the second and third days, and 375 mg/m on the first day of the 21-day cycle of the first cycle Rituximab was administered intravenously at a dose of ² , and in every 21-day cycle after the first cycle, in each 21-day cycle, the immunoconjugate was intravenously at a dose of 1.8 mg/kg intravenously on the first day Administration, bendamustine or its salt was administered intravenously at a dose of 90 mg/m ^{2 on} the first and second days, and rituximab was administered at a dose of 375 mg/m ² on the first day Intravenous administration. In some embodiments, the immunoconjugate and bendamustine or its salt are administered sequentially on the second day of the first cycle. In some embodiments, the immunoconjugate is administered before bendamustine or a salt or solvate thereof. In some embodiments, the immunoconjugate, bendamustine or a salt thereof, and rituximab are administered sequentially on day 1 of cycles 2-6. In some embodiments, rituximab is administered before the immunoconjugate, and wherein the immunoconjugate is administered before the alkylating agent on day 1 of cycles 2-6. In some embodiments, the immunoconjugate, bendamustine or its salt, and rituximab are further administered after the 6th cycle, and wherein in each cycle after the 6th cycle, in each 21 In a daily cycle, the immunoconjugate was administered intravenously at a dose of 1.8 mg/kg on the first day, and bendamustine or its salt was administered intravenously at a dose of 90 mg/m ^{2 on} the first and second days And on the first day, rituximab was administered intravenously at a dose of 375 mg/m ² . In some embodiments, rituximab is administered before the immunoconjugate, and wherein the immunoconjugate is administered before the alkylating agent on day 1 of each 21-day cycle in each cycle after the 6th cycle . Other exemplary dosing and administration regimens are provided elsewhere herein.

In some embodiments, the treatment extends the human PFS to at least about 6, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8. , 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17 or more than 17 months. In some embodiments, treatment extends PFS to at least 7 months. In some embodiments, the treatment extends PFS to at least about 7.6 months. In some embodiments, treatment extends PFS to at least about 8 months. In some embodiments, treatment extends PFS to at least 11 months. In some embodiments, treatment extends PFS to at least 11.1 months.

In some embodiments, the treatment extends the human OS to at least about 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, or more than 12.5 month. In some embodiments, treatment prolongs OS to at least about 12 months. In some embodiments, treatment prolongs OS by at least about 12.4 months.

In some embodiments, DLBCL is an activated B cell-like DLBCL (ABC DLBCL). In some embodiments, DLBCL is germinal center B cell-like DLBCL (GCB DLBCL). In some embodiments, DLBCL (DLBCL-NOS) is not listed. In some embodiments, DLBCL is a dual manifestation lymphoma (DEL). In some embodiments, DLBCL is relapsed/refractory DLBCL. In some embodiments, the human does not have grade 3b follicular lymphoma, transformed and mild non-Hodgkin's lymphoma, or CNS lymphoma. In some embodiments, the human has received at least one previous therapy for DLBCL. In some embodiments, the human has received at least two previous therapies for DLBCL. In some embodiments, the human has received at least three previous therapies for DLBCL. In some embodiments, the person has received more than three previous therapies for DLBCL. In some embodiments, humans are not suitable for autologous stem cell transplantation (ASCT). In some embodiments, the ASCT is first-line ASCT, second-line ASCT, third-line ASCT, or beyond third-line ASCT. In some embodiments, previous autologous stem cell transplantation in humans has failed. In some embodiments, the human has received previous treatment with anti-CD20 agents. In some embodiments, the human has received previous treatment with bendamustine or its salt. In some embodiments, the person is refractory to the most recent prior therapy. In some embodiments, the most recent prior therapy is the standard of care therapy. In some embodiments, if the individual exhibits a partial response, minimal response, or no response to the treatment, the individual is refractory to the treatment. In some embodiments, the individual system is female. In some embodiments, an adult with unspecified (NOS) DLBCL has received at least one previous therapy (eg, for DLBCL).

, 其中Ab係抗CD79b抗體，其包含(i)包含SEQ ID NO：21之胺基酸序列之HVR-H1；(ii)包含SEQ ID NO：22之胺基酸序列之HVR-H2；(iii)包含SEQ ID NO：23之胺基酸序列之HVR-H3；(iv)包含SEQ ID NO：24之胺基酸序列之HVR-L1；(v)包含SEQ ID NO：25之胺基酸序列之HVR-L2；(vi)包含SEQ ID NO：26之胺基酸序列之HVR-L3，並且其中p為1至8，根據本文提供之方法，該抗體與烷化劑及抗CD20抗體組合用於治療有此需要之患有彌漫性大B細胞淋巴瘤(DLBCL)的人。在一些實施例中，抗CD79b抗體包含(i)包含VH之重鏈，該VH包含SEQ ID NO：19之胺基酸序列，及(ii)包含VL之輕鏈，該VL包含SEQ ID NO：20之胺基酸序列。A kit containing the immunoconjugate of the following formula is also provided

, Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iii) ) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25 HVR-L2; (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and wherein p is 1 to 8. According to the method provided herein, the antibody is used in combination with an alkylating agent and an anti-CD20 antibody For the treatment of people with diffuse large B-cell lymphoma (DLBCL) in need. In some embodiments, the anti-CD79b antibody comprises (i) a heavy chain comprising VH, the VH comprising the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, the VL comprising SEQ ID NO: 20 amino acid sequence.

, 其中Ab係抗CD79b抗體，其包含(i)包含VH之重鏈，該VH包含SEQ ID NO：19之胺基酸序列，及(ii)包含VL之輕鏈，該VL包含SEQ ID NO：20之胺基酸序列，並且其中p為2至5，根據本文提供之方法，該抗體與苯達莫司汀或其鹽及利妥昔單抗組合用於治療有此需要之患有彌漫性大B細胞淋巴瘤(DLBCL)的人。在一些實施例中，p介於3與4之間(例如，3.5)。在一些實施例中，重鏈包含SEQ ID NO：36之胺基酸序列，並且其中輕鏈包含SEQ ID NO：35之胺基酸序列。A kit containing the immunoconjugate of the following formula is also provided

, Wherein Ab is an anti-CD79b antibody, which comprises (i) a heavy chain comprising VH, which VH comprises the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, which VL comprises SEQ ID NO: The amino acid sequence of 20, and where p is 2 to 5. According to the method provided herein, the antibody is combined with bendamustine or its salt and rituximab for the treatment of patients with diffuse People with large B-cell lymphoma (DLBCL). In some embodiments, p is between 3 and 4 (eg, 3.5). In some embodiments, the heavy chain comprises the amino acid sequence of SEQ ID NO: 36, and wherein the light chain comprises the amino acid sequence of SEQ ID NO: 35.

, 其中Ab係抗CD79b抗體，其包含(i)包含SEQ ID NO：21之胺基酸序列之HVR-H1；(ii)包含SEQ ID NO：22之胺基酸序列之HVR-H2；(iii)包含SEQ ID NO：23之胺基酸序列之HVR-H3；(iv)包含SEQ ID NO：24之胺基酸序列之HVR-L1；(v)包含SEQ ID NO：25之胺基酸序列之HVR-L2；(vi)包含SEQ ID NO：26之胺基酸序列之HVR-L3，並且其中p在1與8之間，該抗體用於治療有此需要之人之彌漫性大B細胞淋巴瘤(DLBCL)的方法，該方法包括投與人有效量之免疫結合物、烷化劑及抗C20抗體，其中該治療延長人之無進展生存期(PFS)及/或總生存期(OS)。在一些實施例中，免疫結合物用於本文提供之方法中。在一些實施例中，抗CD79b抗體包含(i)包含VH之重鏈，該VH包含SEQ ID NO：19之胺基酸序列，及(ii)包含VL之輕鏈，該VL包含SEQ ID NO：20之胺基酸序列。Also provided is an immunoconjugate comprising the formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iii) ) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25 HVR-L2; (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and wherein p is between 1 and 8. The antibody is used for the treatment of diffuse large B cells in people in need A method for lymphoma (DLBCL), the method comprising administering an effective amount of an immunoconjugate, an alkylating agent and an anti-C20 antibody, wherein the treatment prolongs the progression-free survival (PFS) and/or overall survival (OS ). In some embodiments, immunoconjugates are used in the methods provided herein. In some embodiments, the anti-CD79b antibody comprises (i) a heavy chain comprising VH, the VH comprising the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, the VL comprising SEQ ID NO: 20 amino acid sequence.

, 其中Ab係抗CD79b抗體，其包含(i)包含VH之重鏈，該VH包含SEQ ID NO：19之胺基酸序列，及(ii)包含VL之輕鏈，該VL包含SEQ ID NO：20之胺基酸序列，其中p在2與5之間，該抗體用於治療有需要之人之彌漫性大B細胞淋巴瘤(DLBCL)之方法，該方法包括投與人有效量之(a)免疫結合物，(b)苯達莫司汀或其鹽，及(c)利妥昔單抗，其中免疫結合物以1.8 mg/kg之劑量投與，苯達莫司汀或其鹽以90 mg/m² 之劑量投與，並且利妥昔單抗以375 mg/m² 之劑量投與，並且其中該治療延長了人之無進展生存期(PFS)及/或總生存期(OS)。在一些實施例中，免疫結合物用於本文提供之方法中。在一些實施例中，p介於3與4之間(例如，3.5)。在一些實施例中，抗CD79抗體包含重鏈，該重鏈包含SEQ ID NO：36之胺基酸序列，並且輕鏈包含SEQ ID NO：35之胺基酸序列。Also provided is an immunoconjugate comprising the formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) a heavy chain comprising VH, the VH comprising the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, the VL comprising SEQ ID NO: The amino acid sequence of 20, where p is between 2 and 5. The antibody is used for the treatment of diffuse large B-cell lymphoma (DLBCL) in a person in need. The method comprises administering an effective amount of (a ) Immunoconjugate, (b) bendamustine or its salt, and (c) rituximab, wherein the immunoconjugate is administered at a dose of 1.8 mg/kg, and bendamustine or its salt is The dose of 90 mg/m ² was administered, and rituximab was administered at a dose of 375 mg/m ² , and the treatment prolonged the progression-free survival (PFS) and/or overall survival (OS ). In some embodiments, immunoconjugates are used in the methods provided herein. In some embodiments, p is between 3 and 4 (eg, 3.5). In some embodiments, the anti-CD79 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:36, and the light chain comprising the amino acid sequence of SEQ ID NO:35.

, 其中Ab係抗CD79b抗體，其包含(i)包含SEQ ID NO：21之胺基酸序列之HVR-H1；(ii)包含SEQ ID NO：22之胺基酸序列之HVR-H2；(iii)包含SEQ ID NO：23之胺基酸序列之HVR-H3；(iv)包含SEQ ID NO：24之胺基酸序列之HVR-L1；(v)包含SEQ ID NO：25之胺基酸序列之HVR-L2；(vi)包含SEQ ID NO：26之胺基酸序列之HVR-L3，並且其中p在1與8之間，該抗體用於治療有此需要之人之彌漫性大B細胞淋巴瘤(DLBCL)之方法，該方法包括投與人有效量之組成物、烷化劑及抗C20抗體，其中該治療延長人之無進展生存期(PFS)及/或總生存期(OS)。在一些實施例中，該組成物用於本文提供之方法中。在一些實施例中，抗CD79b抗體包含(i)包含VH之重鏈，該VH包含SEQ ID NO：19之胺基酸序列，及(ii)包含VL之輕鏈，該VL包含SEQ ID NO：20之胺基酸序列。Also provided is a composition (for example, a pharmaceutical composition) comprising an immunoconjugate of the following formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iii) ) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25 HVR-L2; (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and wherein p is between 1 and 8. This antibody is used for the treatment of diffuse large B cells in people in need A method for lymphoma (DLBCL), the method comprising administering an effective amount of a composition, an alkylating agent and an anti-C20 antibody, wherein the treatment prolongs the progression-free survival (PFS) and/or overall survival (OS) of the person . In some embodiments, the composition is used in the methods provided herein. In some embodiments, the anti-CD79b antibody comprises (i) a heavy chain comprising VH, the VH comprising the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, the VL comprising SEQ ID NO: 20 amino acid sequence.

, 其中Ab係抗CD79b抗體，其包含(i)包含VH之重鏈，該VH包含SEQ ID NO：19之胺基酸序列，及(ii)包含VL之輕鏈，該VL包含SEQ ID NO：20之胺基酸序列，其中p在2與5之間，該抗體用於治療有需要之人之彌漫性大B細胞淋巴瘤(DLBCL)之方法，該方法包括投與人有效量之(a)組成物，(b)苯達莫司汀或其鹽，及(c)利妥昔單抗，其中投與該組成物以提供1.8 mg/kg之免疫結合物劑量，苯達莫司汀或其鹽以90 mg/m² 之劑量投與，利妥昔單抗以375 mg/m² 之劑量投與，並且其中治療延長了人之無進展生存期(PFS)及/或總生存期(OS)。在一些實施例中，該組成物用於本文提供之方法中。在一些實施例中，p介於3與4之間(例如，3.5)。在一些實施例中，抗CD79抗體包含有包含胺基酸序列SEQ ID NO: 36之重鏈及包含胺基酸序列SEQ ID NO: 35之輕鏈。Also provided is a composition (for example, a pharmaceutical composition) comprising an immunoconjugate of the following formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) a heavy chain comprising VH, which VH comprises the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, which VL comprises SEQ ID NO: The amino acid sequence of 20, where p is between 2 and 5. The antibody is used for the treatment of diffuse large B-cell lymphoma (DLBCL) in a person in need. The method comprises administering an effective amount of (a ) Composition, (b) bendamustine or its salt, and (c) rituximab, wherein the composition is administered to provide an immunoconjugate dose of 1.8 mg/kg, bendamustine or The salt is administered at a dose of 90 mg/m ² , and rituximab is administered at a dose of 375 mg/m ² , and the treatment prolongs the progression-free survival (PFS) and/or overall survival ( OS). In some embodiments, the composition is used in the methods provided herein. In some embodiments, p is between 3 and 4 (eg, 3.5). In some embodiments, the anti-CD79 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 36 and a light chain comprising the amino acid sequence of SEQ ID NO: 35.

It should be understood that one, some or all of the properties of the various embodiments described herein can be combined to form other embodiments of the invention. These and other aspects of the invention will become apparent to those familiar with the art. These and other embodiments of the present invention are further described by the following detailed description.

Provided herein is a method of treating or delaying the progression of lymphoma (e.g., diffuse large B-cell lymphoma (DLBCL), such as relapsed/refractory DLBCL) in an individual (e.g., human), including administering to the individual an effective amount of anti-CD79b immunity Conjugates, alkylating agents ( e.g. , bendamustine or bendamustine-HCl), and anti-CD20 agents ( e.g., anti-CD20 antibodies, such as rituximab). In some embodiments, treatment with anti-CD79 immunoconjugates, alkylating agents, and anti-CD20 agents prolongs the progression-free survival (PFS) and/or overall survival (OS) of the individual. In some embodiments, such treatment prolongs the progression-free survival (PFS) and/or overall survival (OS) of the individual, for example, and is included in the absence of an anti-CD79 immunoconjugate (e.g., huMA79bv28-MC-vc- In the case of PAB-MMAE or polatuzumab vedotin), PFS of an individual who is administered an alkylating agent (for example, bendamustine or bendamustine-HCl) and an anti-CD20 agent (for example, an anti-CD20 antibody) And/or OS compared. In some embodiments, individuals who are administered anti-CD79 immunoconjugates, alkylating agents, and anti-CD20 agents achieve complete remission (CR) after administration. Complete relief is also called "complete response".

, 其中Ab係抗CD79b抗體，其包含(i)包含SEQ ID NO：21之胺基酸序列之HVR-H1；(ii)包含SEQ ID NO：22之胺基酸序列之HVR-H2；(iii)包含SEQ ID NO：23之胺基酸序列之HVR-H3；(iv)包含SEQ ID NO：24之胺基酸序列之HVR-L1；(v)包含SEQ ID NO：25之胺基酸序列之HVR-L2；(vi)包含SEQ ID NO：26之胺基酸序列之HVR-L3，並且其中p為1至8 (例如，2至5，或3至4)，(b)烷化劑(例如，苯達莫司汀或苯達莫司汀-HCl)，及(c)抗CD20劑(例如利妥昔單抗)，其中免疫結合物以1.8 mg/kg之劑量投與，烷化劑(例如，苯達莫司汀或苯達莫司汀-HCl)以90 mg/m² 之劑量投與，並且以375 mg/m² 之劑量投與抗CD20劑(例如利妥昔單抗)，並且其中該治療延長個人之無進展生存期(PFS)及/或總生存期(OS)。在一些實施例中，投與抗CD79免疫結合物、苯達莫司汀(或苯達莫司汀-HCl)及利妥昔單抗之個體在投與後達成完全緩解(CR)。完全緩解也稱為「完全響應」。In some embodiments, the method comprises treating an individual with diffuse large B-cell lymphoma (DLBCL, such as relapsed/refractory DLBCL) by administering to the individual (a) an immunoconjugate comprising the formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iii) ) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25 HVR-L2; (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and wherein p is 1 to 8 (for example, 2 to 5, or 3 to 4), (b) alkylating agent (For example, bendamustine or bendamustine-HCl), and (c) anti-CD20 agents (for example, rituximab), wherein the immunoconjugate is administered at a dose of 1.8 mg/kg, alkylating Agent (for example, bendamustine or bendamustine-HCl) is administered at a dose of 90 mg/m ² and an anti-CD20 agent (for example, rituximab) is administered at a dose of 375 mg/m ² ), and wherein the treatment prolongs the progression-free survival (PFS) and/or overall survival (OS) of the individual. In some embodiments, individuals who are administered the anti-CD79 immunoconjugate, bendamustine (or bendamustine-HCl), and rituximab achieve complete remission (CR) after administration. Complete relief is also called "complete response".

In some embodiments, the individual has activated B-cell-like DLBCL (ABC DLBCL). In some embodiments, the individual has germinal center B cell-like DLBCL (GCB DLBCL). In some embodiments, the individual has DLBCL not specified (DLBCL-NOS). In some embodiments, the individual has dual manifestation lymphoma (DEL). In some embodiments, the individual does not respond to the initial treatment of DLBCL. In some embodiments, the individual has relapsed/refractory DLBCL. In some embodiments, the individual has received at least one, at least two, or at least three previous therapies for DLBCL. In some embodiments, the individual has received more than three previous therapies for DLBCL. In some embodiments, the individual is not suitable for autologous stem cell transplantation (ASCT) (eg, first-line ASCT, second-line ASCT, third-line ASCT, or beyond third-line ASCT). In some embodiments, the individual's previous autologous stem cell transplantation has failed. In some embodiments, the individual has received previous treatment with an anti-CD20 agent. In some embodiments, the individual has received prior treatment with bendamustine or bendamustine-HCl. In some embodiments, the individual is refractory to the most recent previous therapy. I. General technology

Unless otherwise indicated, the practice of the present invention will use the conventional techniques of molecular biology (including recombinant technology), microbiology, cell biology, biochemistry, and immunology, and these techniques are within the technical scope of this field. These techniques are fully explained in the literature, such as "Molecular Cloning: A Laboratory Manual", second edition (Sambrook et al., 1989); "Oligonucleotide Synthesis" (MJ Gait, ed., 1984); "Animal Cell Culture" (RI Freshney, ed., 1987); "Methods in Enzymology" (Academic Press); "Current Protocols in Molecular Biology" (FM Ausubel et al., 1987 and regularly updated); "PCR: The Polymerase Chain Reaction", (Mullis et al. Edited, 1994); "A Practical Guide to Molecular Cloning" (Perbal Bernard V., 1988); "Phage Display: A Laboratory Manual" (Barbas et al., 2001). II. Definition

Before describing the present invention in detail, it should be understood that the present invention is not limited to specific compositions or biological systems, and they can of course vary. It should be understood that the terminology used herein is for the purpose of describing specific embodiments and is not intended to be limiting.

As used in this specification and the scope of the accompanying application, the singular forms "one" and "the" include plural references unless the content clearly specifies otherwise. Thus, for example, reference to "a molecule" optionally includes a combination of two or more such molecules, and the like.

As used herein, the term "about" refers to the usual error range of individual values that are easily known to those skilled in the art. The reference to "about" a value or parameter in this text includes (and describes) an embodiment that pertains to that value or parameter.

It should be understood that the aspects and embodiments of the present invention described herein include "consisting of aspects and embodiments" and/or "essentially consisting of aspects and embodiments".

Unless otherwise indicated, the term "CD79b" as used herein refers to any natural CD79b from any vertebrate source, including mammals, such as primates ( e.g., humans, cynomolgus monkeys) ( "Cyno")) and rodents ( such as mice and rats). Human CD79b is also referred to herein as "Igβ", "B29", "DNA225786" or "PRO36249". An exemplary CD79b sequence including the signal sequence is shown in SEQ ID NO:1. An exemplary CD79b sequence without a signal sequence is shown in SEQ ID NO:2. The term "CD79b" encompasses "full-length" unprocessed CD79b and any form of CD79b produced by processing in cells. The term also covers naturally occurring variants of CD79b, such as splice variants, allele variants and isoforms. The CD79b polypeptides described herein can be isolated from a variety of sources, such as from a human tissue type or from another source, or prepared by recombinant or synthetic methods. "Native sequence CD79b polypeptide" includes a polypeptide having the same amino acid sequence as the corresponding CD79b polypeptide derived from nature. These native sequence CD79b polypeptides can be isolated from nature, or can be produced by recombinant or synthetic methods. The term "native sequence CD79b polypeptide" specifically includes naturally occurring truncated or secreted forms of a specific CD79b polypeptide (for example, extracellular domain sequence), naturally occurring variant forms of the polypeptide (for example, alternative splicing form) and natural Allelic variants that exist.

"CD20" as used herein refers to the human B lymphocyte antigen CD20 (also known as CD20, B lymphocyte surface antigen B1, Leu-16, Bp35, BM5 and LF5; this sequence is characterized by SwissProt database entry P11836). A hydrophobic transmembrane protein with a molecular weight of approximately 35 kD located on pre-B and mature B lymphocytes. (Valentine, MA et al., J. Biol. Chem. 264(19) (1989 11282-11287; Tedder, TF et al., Proc. Natl. Acad. Sci. USA 85 (1988) 208-12; Stamenkovic, I. Et al., J. Exp. Med. 167 (1988) 1975-80; Einfeld, DA et al., EMBO J. 7 (1988) 711-7; Tedder, TF et al., J. Immunol. 142 (1989) 2560- 8). The corresponding human gene is transmembrane 4 domain, subfamily A, member 1, also known as MS4A1. This gene encodes a member of the transmembrane 4A gene family. The members of the nascent protein family share common structural features and similarities Intron/exon splicing boundary to characterize, and shows a unique expression pattern in hematopoietic cells and non-lymphoid tissues. This gene encodes B lymphocyte surface molecules, which play a role in the development and differentiation of B cells into plasma cells Role. In a cluster of family members, this family member is localized to 11q12. The alternative splicing of the gene produces two transcript variants encoding the same protein.

The terms "CD20" and "CD20 antigen" are used interchangeably herein, and include any variants, isoforms, and species homologs of human CD20, which are naturally expressed by cells or on cells transfected with CD20 gene which performed. The binding of the antibody of the present invention to the CD20 antigen mediates the killing of CD20-expressing cells ( e.g., tumor cells) by inactivating CD20. The killing of CD20-expressing cells can occur by one or more of the following mechanisms: cell death/apoptosis induction, ADCC and CDC. As recognized in this technology, synonyms for CD20 include B lymphocyte antigen CD20, B lymphocyte surface antigen B1, Leu-16, Bp35, BM5 and LF5.

The term "CD20 antigen expression" is intended to mean the significant level of CD20 antigen expression in cells ( such as T- or B-cells). In one embodiment, patients treated according to the methods of the present invention exhibit significant levels of CD20 on B-cell tumors or cancers. Patients with "cancer manifesting CD20" can be determined by standard tests known in this technology. For example , immunohistochemistry (IHC) detection, FACS or PCR-based detection via corresponding mRNA is used to measure CD20 antigen expression.

"Affinity" refers to the total strength of non-covalent interactions between a single binding site of a molecule ( eg , an antibody) and its binding partner ( eg , an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the inherent binding affinity that reflects a 1:1 interaction between members of a binding pair ( eg , antibody and antigen). The affinity of molecule X to its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein. Specific illustrative and exemplary examples for measuring binding affinity are described below.

"Affinity maturation" antibody system refers to an antibody that has one or more changes in one or more hypervariable regions (HVR) compared with the parent antibody. The parent antibody does not have these changes, and the changes result in the antibody pair Improvement of antigen affinity.

The term "antibody" is used in the broadest sense herein and encompasses various antibody structures, including (but not limited to) monoclonal antibodies, multiple antibodies, multispecific antibodies ( such as bispecific antibodies), and antibody fragments, as long as they exhibit The required antigen binding activity is sufficient.

"Antibody fragment" refers to a molecule that is not a complete antibody, which includes the part of a complete antibody that binds to the antigen bound by the complete antibody. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab') ₂ ; bifunctional antibodies; linear antibodies; single-chain antibody molecules ( such as scFv); and many forms of antibody fragments Specific antibodies.

The "antibody that binds to the same epitope" as the reference antibody refers to an antibody that blocks the binding of the reference antibody to its antigen by 50% or more in the competition analysis, and on the contrary, the reference antibody blocks the reference antibody in the competition analysis The binding of the antibody to its antigen is 50% or more. This article provides illustrative competitive analysis.

The term "antigenic determinant" refers to the specific site on the antigen molecule to which the antibody binds.

The term "chimeric" antibody system refers to an antibody in which a part of the heavy chain and/or light chain is derived from a specific source or species, and the remaining part of the heavy chain and/or light chain is derived from a different source or species.

The "class" of an antibody refers to the type of constant domain or constant region possessed by its heavy chain. There are five main classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and some of these classes can be further divided into subclasses (isotypes), such as IgG ₁ , IgG ₂ , IgG ₃ , IgG ₄ , IgA ₁ and IgA ₂ . The heavy chain constant domains corresponding to different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

The terms "anti-CD79b antibody" and "antibody that binds CD79b" refer to antibodies that can bind to CD79b with an affinity sufficient to make the antibody useful as a diagnostic and/or therapeutic agent when targeting CD79b. Preferably, the extent to which the anti-CD79b antibody binds to irrelevant non-CD79b proteins is less than about 10% of the binding of the antibody to CD79b, as measured, for example, by radioimmunoassay (RIA). In certain embodiments, the CD79b-binding antibody has a dissociation constant (Kd) of ≤1 μM, ≤100 nM, ≤10 nM, ≤1 nM, or ≤0.1 nM. In certain embodiments, the anti-CD79b antibody binds to an epitope in CD79b that is conserved among CD79b from different species.

The term "anti-CD20 antibody" according to the present invention refers to an antibody capable of binding to CD20 with an affinity sufficient to make the antibody useful as a diagnostic and/or therapeutic agent when targeting CD20. In one embodiment, the extent to which the anti-CD20 antibody binds to irrelevant non-CD20 proteins is less than about 10% of the binding of the antibody to CD20, as measured, for example, by radioimmunoassay (RIA). In certain embodiments, the CD20-binding antibody has a dissociation constant (Kd) of ≤1 μM, ≤100 nM, ≤10 nM, ≤1 nM, or ≤0.1 nM. In certain embodiments, anti-CD20 antibodies bind to epitopes in CD20 that are conserved among CD20 from different species.

"Isolated" antibodies are antibodies that have been separated from components of their natural environment. In some embodiments, the antibody is purified to greater than 95% or 99% purity, such as by, for example, electrophoresis ( e.g. , SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography ( e.g. , ion exchange or Reverse phase HPLC) determined. For a review of methods used to analyze antibody purity, see, for example, Flatman et al., J. Chromatogr. B 848:79-87 (2007). The "variable region" or "variable domain" of an antibody refers to the amino terminal domain of an antibody heavy or light chain. The variable domain of the heavy chain can be referred to as "VH". The variable domain of the light chain can be called "VL". These domains are usually the most variable part of the antibody and contain the antigen binding site.

"Isolated nucleic acid encoding an anti-CD79b antibody" refers to one or more nucleic acid molecules encoding antibody heavy and light chains (or fragments thereof), including such nucleic acid molecules and their presence in a single vector or separate vectors The nucleic acid molecule(s) at one or more locations in the host cell.

As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, that is, the individual antibodies constituting the population are identical and/or bind to the same epitope, except for possible variants Antibodies, for example, contain naturally occurring mutations or appear during the production of monoclonal antibody preparations, and these variants are generally present in small amounts. In contrast to multiple antibody preparations which typically include different antibodies directed against different determinants (antigenic determinants), each monoclonal antibody of the monoclonal antibody preparation is directed against a single determinant on the antigen. Therefore, the modifier "monoclonal" indicates that the characteristics of the antibody are obtained from a substantially homogeneous population of antibodies, and should not be interpreted as requiring the production of the antibody by any specific method. For example, the monoclonal antibody to be used according to the present invention can be produced by a variety of techniques, including but not limited to fusion tumor methods, recombinant DNA methods, phage display methods, and the use of transgenic animals containing all or part of the human immunoglobulin locus The methods, these methods and other exemplary methods for making monoclonal antibodies are described herein.

"Naked antibody" refers to an antibody that is not bound to a heterogeneous part ( for example , a cytotoxic part) or a radioactive label. The naked antibody may be present in a pharmaceutical formulation.

"Native antibodies" refer to naturally occurring immunoglobulin molecules with varying structures. For example, a native IgG antibody is a heterotetrameric glycoprotein of about 150,000 daltons, composed of two identical light chains and two identical heavy chains bonded by disulfide bonds. From N-terminus to C-terminus, each heavy chain has a variable region (VH) (also called variable heavy domain or heavy chain variable domain), followed by three constant domains (CH1, CH2, and CH3). Similarly, from the N-terminus to the C-terminus, each light chain has a variable region (VL) (also called a variable light domain or a light chain variable domain), followed by a constant light (CL) domain. The light chain of an antibody can be designated as one of two types called kappa and lambda based on the amino acid sequence of its constant domain.

The term "Fc region" herein is used to define the C-terminal region of an immunoglobulin heavy chain that contains at least a part of the constant region. The term includes native sequence Fc region and variant Fc region. In one embodiment, the Fc region of a human IgG heavy chain extends from Cys226 or from Pro230 to the carboxy terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc region may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region is based on the EU numbering system, which is also known as the EU index, such as Kabat et al., Sequences of Proteins of Immunological Interest , 5th edition Described in Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

"Framework" or "FR" refers to variable domain residues that are not hypervariable region (HVR) residues. The FR of a variable domain generally consists of four FR domains: FR1, FR2, FR3, and FR4. Therefore, HVR and FR sequences generally appear in VH (or VL) in the following order: FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

For the purpose of this article, "acceptor human framework" is a light chain variable domain (VL) framework or heavy chain variable domain (VH) framework derived from a human immunoglobulin framework or a human common framework as defined below The framework of the amino acid sequence. The acceptor human framework "derived from" the human immunoglobulin framework or the human common framework may include the same amino acid sequence of the human immunoglobulin framework or the human common framework, or it may contain amino acid sequence changes. In some embodiments, the number of amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 One or less, 3 or less, or 2 or less. In some embodiments, the sequence of the VL acceptor human framework is identical to the VL human immunoglobulin framework sequence or the human common framework sequence.

The terms "full-length antibody", "whole antibody" and "whole antibody" are used interchangeably herein and refer to an antibody whose structure is substantially similar to that of a native antibody or has a heavy chain containing an Fc region as defined herein.

The terms "host cell", "host cell line" and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include "transformants" and "transformed cells", which include primary transformed cells and progeny derived from them regardless of the number of generations. The offspring may not be exactly the same as the parent cell in terms of nucleic acid content, but may contain mutations. This document includes mutant progeny that have the same function or biological activity as that selected or selected for the original transformed cell.

"Human antibodies" are antibodies that have amino acid sequences that correspond to those of antibodies produced by human or human cells or are derived from non-human sources that utilize the human antibody lineage or other human antibody coding sequences. This definition of human antibodies specifically excludes humanized antibodies that contain non-human antigen-binding residues.

"Human common framework" refers to the framework of the amino acid residues most frequently occurring in the selected human immunoglobulin VL or VH framework sequence. In general, the human immunoglobulin VL or VH sequence is selected from a subgroup of variable domain sequences. Generally speaking, the sequence subgroup is as in Kabat et al., Sequences of Proteins of Immunological Interest , Fifth Edition, NIH Publication 91-3242, Bethesda MD (1991), Vols 1-3. In one embodiment, for VL, the subgroup is subgroup κI as in Kabat et al. (supra). In one embodiment, for VH, the subgroup is subgroup III as in Kabat et al. (supra).

"Humanized" antibodies refer to chimeric antibodies containing amino acid residues from non-human HVR and amino acid residues from human FR. In certain embodiments, the humanized antibody will comprise substantially all of at least one and typically two variable domains, wherein all or substantially all of the HVR (e.g., CDR) corresponds to those of the non-human antibody, and All or substantially all FRs correspond to those of human antibodies. The humanized antibody may optionally comprise at least a part of the constant region of an antibody derived from a human antibody. The "humanized form" of an antibody (eg, a non-human antibody) refers to an antibody that has undergone humanization.

The term "hypervariable region" or "HVR" as used herein refers to each region of an antibody variable domain that is hypervariable in sequence and/or forms a structurally defined loop ("hypervariable loop"). Generally speaking, a natural four-chain antibody contains six HVRs; three in VH (H1, H2, H3), and three in VL (L1, L2, L3). HVR usually contains amino acid residues from hypervariable loops and/or amino acid residues from the "complementarity determining region" (CDR). The complementarity determining region has the highest sequence variability and/or is involved in antigen recognition. Exemplary hypervariable rings exist in amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2) and 96-101 (H3). (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987).) Exemplary CDRs (CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3) exist Amino acid residues 24-34 in L1, amino acid residues 50-56 in L2, amino acid residues 89-97 in L3, amino acid residues 31-35B in H1, amino acid residues in H2 Residues 50-65 and H3 amino acid residues 95-102. (Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition, Public Health Service, National Institutes of Health, Bethesda, MD (1991).) In addition to CDR1 in VH, CDR usually contains an amine that forms a hypervariable ring Base acid residues. CDRs also include "specificity determining residues" or "SDRs" as residues that contact the antigen. SDR is contained in a region of CDR called abbreviated-CDR (abbreviated-CDR) or a-CDR. Exemplary a-CDR (a-CDR-L1, a-CDR-L2, a-CDR-L3, a-CDR-H1, a-CDR-H2 and a-CDR-H3) are present in the amino acid residues of L1 Group 31-34, L2 amino acid residue 50-55, L3 amino acid residue 89-96, H1 amino acid residue 31-35B, H2 amino acid residue 50-58 and H3 The amino acid residues 95-102. ( See Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008).) Unless otherwise indicated, HVR residues and other residues ( e.g. , FR residues) in the variable domain are described herein according to Kabat et al. Person, number above.

The term "variable region" or "variable domain" refers to the domain of an antibody heavy or light chain involved in the binding of the antibody to the antigen. The variable domains of the heavy chain and light chain (VH and VL, respectively) of native antibodies generally have similar structures, and each domain contains four conserved framework regions (FR) and three hypervariable regions (HVR). ( See, for example, Kindt et al. Kuby Immunology , 6th edition, WH Freeman and Co., page 91 (2007).) A single VH or VL domain may be sufficient to confer antigen binding specificity. In addition, antibodies that bind to a specific antigen can be isolated using VH or VL domains from antibodies that bind the antigen to screen a library of complementary VL or VH domains, respectively. See, for example , Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

"Effector functions" refer to their biological activities attributable to the Fc region of antibodies, which vary with antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; cell surface receptors (such as B cell receptors) ) Down-regulation; and B cell activation.

"CD79b polypeptide variant" refers to the full-length native sequence CD79b polypeptide sequence disclosed herein, the CD79b polypeptide sequence lacking the signal peptide disclosed herein, the extracellular domain of the CD79b polypeptide disclosed herein with or without the signal peptide, or herein Any other fragments of the disclosed full-length CD79b polypeptide sequence (e.g., those encoded by a nucleic acid that represents only a portion of the complete coding sequence of the full-length CD79b polypeptide) have at least about 80% amino acid sequence identity as defined herein The CD79b polypeptide is preferably an active CD79b polypeptide. Such CD79b polypeptide variants include, for example, a CD79b polypeptide in which one or more amino acid residues are added or deleted at the N- or C-terminus of the full-length native amino acid sequence. Generally, the CD79b polypeptide variant will be the same as the full-length native sequence CD79b polypeptide sequence disclosed herein, the CD79b polypeptide sequence lacking the signal peptide disclosed herein, the extracellular domain of the CD79b polypeptide disclosed herein with or without the signal peptide, or as disclosed herein Any other fragment of the full-length CD79b polypeptide sequence has at least about 80% amino acid sequence identity, or at least about 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89 %, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% amino acid sequence identity. Generally, CD79b variant polypeptides are at least about 10 amino acids long, or at least about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170 , 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420 ,430,440,450,460,470,480,490,500,510,520,530,540,550,560,570,580,590,600 amino acids long, or longer. Optionally, the CD79b variant polypeptide will have no more than one conservative amino acid substitution when compared to the native CD79b polypeptide sequence, or no more than about 2, 3, 4, 5, 6, 7, compared to the native CD79b polypeptide sequence. 8, 9, or 10 conservative amino acid substitutions.

The "percent (%) amino acid sequence identity" of the reference polypeptide sequence is defined as the sequence identity after aligning candidate sequences and introducing gaps (if necessary) to achieve the maximum percent sequence identity. For any conservative substitution of a part, the percentage of amino acid residues in the candidate sequence that are identical to the amino acid residues in the reference polypeptide sequence. It can be aligned in a variety of ways within the skill of this technology, such as using publicly available computer software, such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software, to determine the percent identity of amino acid sequences. Those skilled in the art can determine the appropriate parameters for aligning sequences, including any algorithms required to achieve maximum alignment over the full length of the sequences being compared. However, for the purposes of this article, the sequence comparison computer program ALIGN-2 was used to generate% amino acid sequence identity values. The ALIGN-2 sequence comparison computer program was created by Genentech, and the source code has been archived by user files of the U.S. Copyright Office, Washington D.C., 20559, where it is registered under the U.S. Copyright Registration No. TXU510087. The ALIGN-2 program is publicly available from Genentech, South San Francisco, California, or can be compiled from source code. The ALIGN-2 program should be compiled for UNIX operating system, including digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and remain unchanged.

In the case where ALIGN-2 is used for amino acid sequence comparison, the given amino acid sequence A is relative to, with or against the% amino acid sequence identity of the given amino acid sequence B (which can either be expressed as having or containing Relative, and or against a certain% amino acid sequence identity of a predetermined amino acid sequence B) is calculated as follows: 100×fraction X/Y Where X is the number of amino acid residues scored as unanimous matches by the program in the A and B alignment of the sequence alignment program ALIGN-2, and Y is the total number of amino acid residues in B. It should be understood that when the length of the amino acid sequence A is not equal to the length of the amino acid sequence B, the% amino acid sequence identity of A relative to B will not be equal to the% amino acid sequence identity of B relative to A. Unless specifically stated otherwise, all% amino acid sequence identity values used herein are obtained using the ALIGN-2 computer program as described in the previous paragraph.

As used herein, the term "vector" refers to a nucleic acid molecule that can propagate another nucleic acid molecule to which it is linked. The term includes vectors that are autonomously replicating nucleic acid structures as well as vectors that are embedded in the genome of a host cell into which it has been introduced. Certain vectors can direct the performance of operably linked nucleic acids. Such vectors are referred to herein as "performance vectors".

An "immunoconjugate" is an antibody that binds to one or more heterologous molecules (including but not limited to cytotoxic agents). \

In the context of the formula provided herein, "p" refers to the average number of drug moieties per antibody, which can be in the range of, for example, about 1 to about 20 drug moieties per antibody, and in certain embodiments , 1 to about 8 drug parts per antibody. The present invention includes a composition comprising a mixture of an antibody-drug compound of Formula I, wherein the average drug loading of each antibody is about 2 to about 5, or about 3 to about 4, (for example, about 3.5).

As used herein, the term "cytotoxic agent" refers to a substance that inhibits or prevents cell function and/or causes cell death or destruction. Cytotoxic agents include but are not limited to radioisotopes ( such as At ²¹¹ , I ¹³¹ , I ¹²⁵ , Y ⁹⁰ , Re ¹⁸⁶ , Re ¹⁸⁸ , Sm ¹⁵³ , Bi ²¹² , P ³² , Pb ²¹² and radioactive isotopes of Lu); chemotherapeutic agents Or drugs ( such as methotrexate, adriamycin, vinblastine alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, nitrophenylbutyrate Mustard, daunorubicin or other intercalating agents); growth inhibitors; enzymes and fragments thereof, such as nucleolytic enzymes; antibiotics; toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including Fragments and/or variants; and various anti-tumor or anti-cancer agents disclosed below.

The terms "cancer" and "cancerous" refer to or describe a physiological condition in mammals, which is typically characterized by unregulated cell growth. Examples of cancer include, but are not limited to, B-cell lymphoma (including low-grade/follicular non-Hodgkin’s lymphoma (NHL); small lymphocyte (SL) NHL; intermediate/follicular NHL; intermediate-grade diffuse NHL; high-grade immunity Blastic NHL; high-grade lymphoblastic NHL; high-grade small non-dividing cell NHL; giant mass disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom macroglobulinemia); chronic lymphocytic leukemia (CLL) Acute lymphocytic leukemia (ALL); hairy cell leukemia; chronic myeloblastic leukemia; and post-transplant lymphoproliferative disease (PTLD), as well as keloidosis, edema (such as edema associated with brain tumors) and Meigs syndrome Related abnormal vascular proliferation. More specific examples include, but are not limited to, relapsed or refractory NHL, first-line low-grade NHL, stage III/IV NHL, chemotherapy-resistant NHL, precursor B lymphoblastic leukemia and/or lymphoma, small lymphocytic lymphoma, B cell Chronic lymphocytic leukemia and/or prolymphocytic leukemia and/or small lymphocytic lymphoma, B-cell prolymphocytic lymphoma, immunocytoma and/or lymphoplasmacytic lymphoma, lymphoplasmacytic lymphoma Tumor, marginal zone B-cell lymphoma, splenic marginal zone lymphoma, extranodal marginal zone-MALT lymphoma, nodular marginal zone lymphoma, hairy cell leukemia, plasmacytoma and/or plasma cell myeloma, low grade/follicle Lymphoma, intermediate/follicular NHL, mantle cell lymphoma, follicular center lymphoma (follicular), intermediate diffuse NHL, diffuse large B-cell lymphoma (DLBCL), aggressive NHL (including invasive First-line NHL and aggressive recurrence NHL), NHL that relapsed after autologous stem cell transplantation or refractory to autologous stem cell transplantation, primary mediastinal large B-cell lymphoma, primary exudative lymphoma, high-grade immunoblasts NHL, high-grade lymphoblastic NHL, high-grade small non-dividing cell NHL, huge mass disease NHL, Burkitt’s lymphoma, precursor (peripheral) large granular lymphocytic leukemia, mycosis fungoides and/or seiza Sezary syndrome, cutaneous (cutaneous) lymphoma, degenerative large cell lymphoma, vascular center lymphoma.

"Individual" or "subject" is a mammal. Mammals include, but are not limited to, domesticated animals ( such as cows, sheep, cats, dogs, and horses), primates ( such as humans and non-human primates, such as monkeys), rabbits, and rodents ( such as small animals). Rats and rats). In certain embodiments, the individual or subject is a human.

The "effective amount" of a medicament (such as a pharmaceutical formulation) refers to the amount that is effective to achieve the desired treatment or prevention result at the necessary dose for a necessary period of time.

The term "pharmaceutical formulation" refers to a formulation that allows the biological activity of the active ingredients contained therein to be effective, and does not contain additional components that have unacceptable toxicity to the individual to whom the formulation will be administered.

"Pharmaceutically acceptable carrier" refers to the ingredients in the pharmaceutical formulations other than the active ingredients, which are non-toxic to the individual. Pharmaceutically acceptable carriers include but are not limited to buffers, excipients, stabilizers or preservatives.

As used herein, "treatment" (and its grammatical variations, such as "treat" or "treating") refers to clinical intervention in an attempt to change the natural course of the individual being treated, and can be achieved Prevention or in the course of clinical pathology. Desirable therapeutic effects include but are not limited to reducing free light chains, preventing the occurrence or recurrence of the disease, alleviating symptoms, reducing any direct or indirect pathological results of the disease, reducing the rate of disease progression, improving or alleviating the disease state, and alleviating or improving the prognosis . In some embodiments, the antibodies described herein are used to delay disease production or slow the progression of disease.

The term "CD79b positive cancer" refers to cancers that include cells that express CD79b on the surface. In some embodiments, the expression of CD79b on the cell surface is determined using CD79b antibody in methods such as immunohistochemistry, FACS, and the like. Alternatively, it is believed that the expression of CD79b mRNA is related to the expression of CD79b on the cell surface and can be determined by a method selected from in situ hybridization and RT-PCR (including quantitative RT-PCR).

As used herein, "in combination with" refers to the administration of one form of treatment in addition to another form of treatment. Therefore, "in combination with" refers to the administration of one form of treatment before, during or after the administration of another form of treatment to the individual.

"Chemotherapeutic agents" are compounds suitable for the treatment of cancer. Examples of chemotherapeutics include erlotinib (TARCEVA ^® , Genentech/OSI Pharm.), bortezomib (VELCADE ^® , Millennium Pharm.), disulfiram (disulfiram), epigallium Catechin gallate (epigallocatechin gallate), salinosporamide A (salinosporamide A), carfilzomib (carfilzomib), 17-AAG (geldanamycin), radicicol (radicicol), lactate dehydrogenase A (LDH-A), fulvestrant (FASLODEX ^® , AstraZeneca), sunitinib (SUTENT ^® , Pfizer/Sugen), letrozole ) (FEMARA ^® , Novartis), imatinib mesylate (GLEEVEC ^® , Novartis), finasunate (VATALANIB ^® , Novartis), oxaliplatin (ELOXATIN ^® , Sanofi), 5-FU (5-fluorouracil), leucovorin, Rapamycin (Sirolimus, RAPAMUNE ^® , Wyeth), Lapatinib (TYKERB ^® , GSK572016, Glaxo Smith Kline), Lonafamib (SCH 66336), Sorafenib (NEXAVAR ^® , Bayer Labs), Gefitinib (IRESSA ^® , AstraZeneca), AG1478 , Alkylating agents, such as thiotepa and CYTOXAN ^® cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; nitrogen Propidium, such as benzodopa, carboquone, metedopa and uredopa; ethyleneimine and methylmelamine, including altretamine, Triethylene melamine, triethylene phosphatidamide, tris ethylene thiophosphatidamide and trimethylol melamine; poly acetogenins (especially Bullatacin and bullatacinone); camptothecin (including topotecan and irinotecan); bryostatin; calllystatin ); CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycin (specifically prednisone ( prednisone and prednisolone); cyproterone acetate; 5α-reductase, including finasteride and dutasteride); vorinostat ( vorinostat, romidepsin, panobinostat, valproic acid, mocetinostat, dolastatin; aldesleukin, talc times Duocarmycin (including synthetic analogues KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; spongistatin; nitrogen Mustards, such as chlorambucil, chlomaphazine, chlorophosphamide, estramustine, ifosfamide, dichloromethyldiethyl Amine (mechlorethamine), dichloromethyldiethylamine oxide hydrochloride, melphalan (melphalan), novembichin (novembichin), phenesterine (phenesterine), prednimustine (prednimustine), chlorine Trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine ( lomustine), nimustine and ranimnustine; antibiotics, such as enediyne antibiotics (e.g. calicheamicin, especially calicheamicin γ1I and calicheamicin) ω1I ( Angew Chem. Int l. Ed. Engl. 1994 33:183-186); dynemicin (dynemicin), including danemycin A; bisphosphonates, such as clodronate (clodronate); esperamicin ( esperamicin); and neocarzinostatin (neocarzinostatin) chromophore and related chromoprotein (chromoprotein) enediyne antibiotic chromophore), aclacinomysin, actinomycin, and atoxin ( authramycin), azaserine (azaserine), bleomycin (bleomycin), actinomycin C (cactinomycin), carabicin (carabicin), caminomycin (caminomycin), carzinophilin (carzinophilin), Chromomycinis (chromomycinis), actinomycin D (dactinomycin), daunorubicin (daunorubicin), detorubicin (detorubicin), 6-diazo-5-pendoxy-L-nor-leucine, ADRIAMYCIN ^® (doxorubicin), N-morpholinyl adriamycin, cyano (N-morpholinyl) adriamycin, 2-(N-pyrrolyl) adriamycin and deoxydoxorubicin (deoxydoxorubicin), epirubicin (epirubicin), esorubicin (esorubicin), everolimus (everolimus), sotrataurin (sotrataurin), idarubicin (idarubicin), marcellomycin (marcellomycin) ), mitomycin (such as mitomycin C), mycophenolic acid, nogalamycin, olivomycin, peplomycin, wave Phhromycin (porfiromycin), puromycin (puromycin), tri-iron adriamycin (quelamycin), rhodoubicin (rodorubicin), streptomycin (streptonigrin), streptozocin, tuberculosis Tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites, such as methotrexate and 5-fluorouracil (5-FU); Folic acid analogs, such as denopterin, methotrexate, pteropterin, trimetrexate; purine Pyrimidine analogs, such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs, such as ancitabine, a Azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine , Enocitabine, floxuridine; androgens, such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane ), testolactone (testolactone); antiadrenaline, such as aminoglutethimide, mitotane, trilostane; folic acid supplements, such as frolinic acid ; Acetyl glucaldehyde ester (aceglatone); aldophosphamide glycoside (aldophosphamide glycoside); aminolevulinic acid (aminolevulinic acid); eniluracil (eniluracil); amsacrine (amsacrine); bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elfomithine Ammonium (elliptinium acetate); epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; similar to beauty Maytansinoid, such as maytansine and ansamitocin; mitoguazone; mitoxantrone; mopidamnol; diamine nitrate Nitraerine; pentostatin; phenamet; pirarubicin ; Losoxantrone (losoxantrone); podophyllinic acid; 2-ethylhydrazide (2-ethylhydrazide); procarbazine; PSK ^® polysaccharide complex (JHS Natural Products, Eugene, Oreg. ); razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2, 2',2"-Trichlorotriethylamine; trichothecene (especially T-2 toxin, verracurin A), roridin A and serpentine ( anguidine)); urethan; vindesine; dacarbazine; mannitol mustard (mannomustine); dibromomannitol (mitobronitol); dibromodurol (mitolactol); Pipobroman; gacytosine; arabinoside ("Ara-C");cyclophosphamide;thiotepa; taxoid, such as taxol (TAXOL) (Paclitaxel; Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE ^® (Cremophor-free), paclitaxel albumin engineered nanoparticle formulation (American Pharmaceutical Partners, Schaumberg, Ill.) and TAXOTERE ^® (docetaxel/doxetaxel; Sanofi-Aventis); chloranmbucil; GEMZAR ^® (gemcitabine); 6-thioguanine ( 6-thioguanine); mercaptopurine; methotrexate; platinum analogues such as cisplatin and carboplatin; vinblastine; etoposide (VP-16) ); ifosfamide; mitoxantrone; vincristine (vincristine); NAVELBINE ^® (vinorelbine (vi norelbine)); novantrone; teniposide; edatrexate; daunorubicin; aminopterin; capecitabine (XELODA ^® ); ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoid, such as Retinoic acid; and pharmaceutically acceptable salts, acids and derivatives of any of the above; and combinations of two or more of the above, such as CHOP, cyclophosphamide, doxorubicin, The abbreviation for the combination therapy of vincristine and prednisolone, and the abbreviation for the combination therapy of FOLFOX, oxaliplatin (ELOXATIN ^TM ) and 5-FU and leucovorin. Additional examples include chemotherapeutics, including bendamustine (or bendamustine-HCl) (TREANDA®), ibrutinib, lenalidomide, and/or idelaris (GS-1101) .

Additional examples of chemotherapeutic agents include antihormonal agents used to regulate, reduce, block, or inhibit the effects of hormones that promote cancer growth, and are usually in the form of systemic or whole-body treatment. It can be a hormone by itself. Examples include anti-estrogen and selective estrogen receptor modulators (SERM), including, for example, tamoxifen (including NOLVADEX® tamoxifen), raloxifene (EVISTA®), droloxifene, 4- Hydroxytamoxifen, Travoxifene, Cxifen, LY117018, Onlastone and Toremifene (FARESTON®); Antiprogesterone; Estrogen receptor downregulator (ERD); Estrogen receptor antagonist Agents, such as Fulvestrant (FASLODEX®); agents used to suppress or shut down the ovaries, such as luteinizing hormone (LHRH) agonists, such as leuprolide acetate (LUPRON® and ELIGARD®), gosere acetate Lin, buserelin acetate, and triptorelin; antiandrogens, such as flutamide, nilutamide, and bicalutamide; and aromatase inhibitors that inhibit aromatase that regulates estrogen production in the adrenal glands, such as 4(5)-imidazole, amiluminide, megestrol acetate (MEGASE®), exemestane (AROMASIN®), formestan (formestanie), fadrozole, vorcloazole (RIVISOR®), Letrozole (FEMARA®) and Anastrozole (ARIMIDEX®). In addition, this definition of chemotherapeutics includes bisphosphonates, such as clodronate (for example, BONEFOS® or OSTAC®), etidronate (DIDROCAL®), NE-58095, zoledronic acid/zoledronate Phosphonate (ZOMETA®), Alendronate (FOSAMAX®), Pamidronate (AREDIA®), Tiludronate (SKELID®) or Risedronate (ACTONEL®); and Satabine (1,3-dioxolane cytosine analogue); antisense oligonucleotides, especially those that inhibit gene expression in signal pathways related to abnormal cell proliferation , Such as PKC-α, Raf, H-Ras and epidermal growth factor receptor (EGF-R); vaccines such as THERATOPE® vaccine and gene therapy vaccines, such as ALLOVECTIN® vaccine, LEUVECTIN® vaccine and VAXID® vaccine.

In some embodiments, chemotherapeutic agents include topoisomerase 1 inhibitors ( e.g. , LURTOTECAN®); anti-estrogens, such as fulvestrant; Kit inhibitors, such as imatinib or EXEL-0862 (tyrosine Kinase inhibitors); EGFR inhibitors such as erlotinib or cetuximab; anti-VEGF inhibitors such as bevacizumab; arinotecan; rmRH ( for example , ABARELIX®); lapatinib And lapatinib xylene sulfonate (ErbB-2 and EGFR tyrosine kinase small molecule inhibitor, also known as GW572016); 17AAG (geldanamycin derivative, heat shock protein (Hsp) 90 Toxins), and pharmaceutically acceptable salts, acids and derivatives of any of the above.

Chemotherapeutic agents also include antibodies, such as Alemtuzumab (Campath), Bevacizumab (AVASTIN®, Genentech); Cetuximab (ERBITUX®, Imclone); Panitumumab (VECTIBIX®, Amgen), Rituximab (RITUXAN®, Genentech/Biogen Idec), ublituximab, Ofatumumab, Irituximab, Pertuzumab (OMNITARG®, 2C4, Genentech), Trastuzumab (HERCEPTIN) ®, Genentech), tositumomab (Bexxar, Colixia) and antibody drug conjugates, gemtuzumab ozogamicin (MYLOTARG®, Wyeth). Other humanized monoclonal antibodies that have therapeutic potential as a drug in combination with the compound include: apolizumab, aselizumab, tocilizumab, bapiizumab Anti (bapineuzumab), bivacizumab (bivatuzumab mertansine), mocantuzumab (cantuzumab mertansine), cedelizumab (cedelizumab), pegylated certolizumab pegol (certolizumab pegol), cidfusituzumab, cidtuzumab, daclizumab, eculizumab, efalizumab, epratuzumab, erlizumab, famvirizumab Anti-felvizumab, fontolizumab, gemtuzumab ozogamicin, inotuzumab ozogamicin, ipilimumab, labetuzumab labetuzumab, lintuzumab, matuzumab, mepolizumab, motavizumab, motovizumab, natal Natalizumab, nimotuzumab, nolovizumab, numavizumab, ocrelizumab, omalizumab, palivizumab, pascolizumab, pecfusituzumab, Pertuzumab (pectuzumab), pexelizumab (pexelizumab), palivizumab (ralivizumab), ranibizumab (ranibizumab), relizumab (reslivizumab), relizumab ( reslizumab), resyvizumab, rovelizumab, ruplizumab, sibrotuzumab, siplizumab, sotuzumab, tatuzumab ( tacatuzumab tetraxetan), tadocizumab, talizumab, tefibazumab b), tocilizumab (tocilizumab), toralizumab (toralizumab), Tucotuzumab celmoleukin (tucotuzumab celmoleukin), tucusituzumab, umavizumab, urtoxazumab, ustekinumab, Visilizumab and anti-interleukin-12 (ABT-874/J695, Wyeth Research and Abbott Laboratories), which is a recombinant human-specific full-length IgG1 λ antibody, which has been genetically modified to recognize Baisu-12 p40 protein.

The term "package insert" is used to refer to the instructions customarily included in the commercial packaging of therapeutic products, which contain warnings about indications, usage, dosage, administration, combination therapy, contraindications and/or the use of such therapeutic products的信息。 Information.

"Alkyl" refers to a C ₁ -C ₁₈ hydrocarbon containing normal chain carbon atoms, secondary carbon atoms, tertiary carbon atoms or ring carbon atoms. Alkyl group methyl (Me, -CH ₃ ), ethyl (Et, -CH ₂ CH ₃ ), 1-propyl (n-Pr, n-propyl, -CH ₂ CH ₂ CH ₃ ), 2-propane Group (i-Pr, isopropyl, -CH(CH ₃ ) ₂ ), 1-butyl (n-Bu, n-butyl, -CH ₂ CH ₂ CH ₂ CH ₃ ), 2-methyl-1- Propyl (i-Bu, isobutyl, -CH ₂ CH(CH ₃ ) ₂ ), 2-butyl (s-Bu, second butyl, -CH(CH ₃ )CH ₂ CH ₃ ), 2- Methyl-2-propyl (t-Bu, tertiary butyl, -C(CH ₃ ) ₃ ), 1-pentyl (n-pentyl, -CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2- Pentyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₃ ), 3-pentyl (-CH(CH ₂ CH ₃ ) ₂ ), 2-methyl-2-butyl(-C(CH ₃ ) ₂ CH ₂ CH ₃ ), 3-methyl-2-butyl (-CH(CH ₃ )CH(CH ₃ ) ₂ ), 3-methyl-1-butyl (-CH ₂ CH ₂ CH(CH ₃ ) ₂ ), 2-methyl-1-butyl (-CH ₂ CH(CH ₃ )CH ₂ CH ₃ ), 1-hexyl (-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-hexyl ( -CH(CH ₃ )CH ₂ CH ₂ CH ₂ CH ₃ ), 3-hexyl (-CH(CH ₂ CH ₃ )(CH ₂ CH ₂ CH ₃ )), 2-methyl-2-pentyl(-C (CH ₃ ) ₂ CH ₂ CH ₂ CH ₃ ), 3-methyl-2-pentyl (-CH(CH ₃ )CH(CH ₃ )CH ₂ CH ₃ ), 4-methyl-2-pentyl ( -CH(CH ₃ )CH ₂ CH(CH ₃ ) ₂ ), 3-methyl-3-pentyl (-C(CH ₃ )(CH ₂ CH ₃ ) ₂ ), 2-methyl-3-pentyl (-CH(CH ₂ CH ₃ )CH(CH ₃ ) ₂ ), 2,3-dimethyl-2-butyl (-C(CH ₃ ) ₂ CH(CH ₃ ) ₂ ), 3,3-di Methyl-2-butyl (-CH(CH ₃ )C(CH ₃ ) ₃ .

The term "C ₁ -C ₈ alkyl" as used herein refers to a straight or branched chain saturated or unsaturated hydrocarbon having 1 to 8 carbon atoms. Representative "C ₁ -C ₈ alkyl groups" include (but are not limited to) -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, -n-hexyl, -n-heptyl,- N-octyl, -n-nonyl and -n-decyl; and branched C ₁ -C ₈ alkyl groups include (but are not limited to) -isopropyl, -second butyl, -isobutyl, -tertiary butyl Group, -isopentyl, 2-methylbutyl, unsaturated C ₁ -C ₈ alkyl including (but not limited to) -vinyl, -allyl, -1-butenyl, -2-butene Group, -isobutenyl, -1-pentenyl, -2-pentenyl, -3-methyl-1-butenyl, -2-methyl-2-butenyl, -2,3-di Methyl-2-butenyl, 1-hexyl, 2-hexyl, 3-hexyl, -ethynyl, -propynyl,-1-butynyl,-2-butynyl,-1-pentynyl ,-2-pentynyl, -3-methyl-1-butynyl. The C ₁ -C ₈ alkyl group may be unsubstituted or substituted with one or more groups including (but not limited to) the following: -C ₁ -C ₈ alkyl, -O-(C ₁ -C ₈ alkyl) , -Aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH ₂ , -C(O)NHR', -C(O) N(R') ₂ -NHC(O)R', -SO ₃ R', -S(O) ₂ R', -S(O)R', -OH, -halogen, -N ₃ , -NH ₂ , -NH(R'), -N(R') ₂ and -CN; wherein each R'is independently selected from H, -C ₁ -C ₈ alkyl and aryl.

The term "C ₁ -C ₁₂ alkyl" as used herein refers to a straight or branched chain saturated or unsaturated hydrocarbon having 1 to 12 carbon atoms. The C ₁ -C ₁₂ alkyl group may be unsubstituted or substituted with one or more groups including but not limited to: -C ₁ -C ₈ alkyl, -O-(C ₁ -C ₈ alkyl),- Aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH ₂ , -C(O)NHR', -C(O)N( R') ₂ -NHC(O)R', -SO ₃ R', -S(O) ₂ R', -S(O)R', -OH, -halogen, -N ₃ , -NH ₂ ,- NH(R'), -N(R') ₂ and -CN; wherein each R'is independently selected from H, -C ₁ -C ₈ alkyl and aryl.

The term "C ₁ -C ₆ alkyl" as used herein refers to a straight or branched chain saturated or unsaturated hydrocarbon having 1 to 6 carbon atoms. Representative "C ₁ -C ₆ alkyl groups" include (but are not limited to) -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl and -n-hexyl; and branched C _1- C ₆ alkyl groups include (but are not limited to) -isopropyl, -second butyl, -isobutyl, -tertiary butyl, -isopentyl and 2-methylbutyl; unsaturated C ₁ -C ₆ Alkyl groups include (but are not limited to) -vinyl, -allyl, -1-butenyl, -2-butenyl and -isobutenyl, -1-pentenyl, -2-pentenyl, -3-methyl-1-butenyl, -2-methyl-2-butenyl, -2,3-dimethyl-2-butenyl, 1-hexyl, 2-hexyl and 3-hexyl . The C ₁ -C ₆ alkyl group may be unsubstituted or substituted with one or more groups as described above for the C ₁ -C ₈ alkyl group.

The term "C ₁ -C ₄ alkyl" as used herein refers to a straight or branched chain saturated or unsaturated hydrocarbon having 1 to 4 carbon atoms. Representative "C ₁ -C ₄ alkyl groups" include (but are not limited to) -methyl, -ethyl, -n-propyl, -n-butyl; and branched C ₁ -C ₄ alkyl groups include (but are not limited to) ) -Isopropyl, -second butyl, -isobutyl, -tertiary butyl; unsaturated C ₁ -C ₄ alkyl includes (but not limited to) -vinyl, -allyl, -1- Butenyl, -2-butenyl and -isobutenyl. The C ₁ -C ₄ alkyl group may be unsubstituted or substituted with one or more groups as described above for the C ₁ -C ₈ alkyl group.

The "hydrocarbyloxy group" is an alkyl group bonded to oxygen with a single bond. Exemplary alkoxy groups include, but are not limited to, methoxy (-OCH ₃ ) and ethoxy (-OCH ₂ CH ₃ ). The "C ₁ -C ₅ hydrocarbonoxy group" is a hydrocarbonoxy group having 1 to 5 carbon atoms. The hydrocarbyloxy group may be unsubstituted or substituted with one or more groups as described above for the alkyl group.

"Alkenyl" is a C ₂ -C ₁₈ hydrocarbon with at least one site of unsaturation (i.e. carbon-carbon sp ² double bond) and containing normal chain carbon atoms, secondary carbon atoms, tertiary carbon atoms or ring carbon atoms . Examples include (but are not limited to): vinyl (ethylene/vinyl; -CH=CH ₂ ), allyl (-CH ₂ CH=CH ₂ ), cyclopentenyl (-C ₅ H ₇ ) and 5-hexyl Alkenyl (-CH ₂ CH ₂ CH ₂ CH ₂ CH=CH ₂ ). "C ₂ -C ₈ alkenyl group" has at least one unsaturation site (that is, carbon-carbon sp ² double bond) and contains 2 to 8 normal chain carbon atoms, secondary carbon atoms, tertiary carbon atoms or rings Hydrocarbon of carbon atoms.

"Alkynyl" is a C ₂ -C ₁₈ hydrocarbon with at least one unsaturation site (ie, carbon-carbon sp- bond) and containing normal chain carbon atoms, secondary carbon atoms, tertiary carbon atoms or ring carbon atoms. Examples include (but are not limited to): ethynyl (-C≡CH) and propargyl (-CH ₂ C≡CH). "C ₂ -C ₈ alkynyl group" has at least one site of unsaturation (i.e. carbon-carbon sp parameter bond) and contains 2 to 8 normal chain carbon atoms, secondary carbon atoms, tertiary carbon atoms or ring carbons Atomic hydrocarbon.

"Alkylene" refers to a saturated branched chain having 1-18 carbon atoms and having two monovalent group centers obtained by removing two hydrogen atoms from the same carbon atom or two different carbon atoms of the parent alkane Or linear or cyclic alkyl. Typical alkylene groups include but are not limited to: methylene (-CH ₂ -), 1,2-ethyl (-CH ₂ CH ₂ -), 1,3-propyl (-CH ₂ CH ₂ CH ₂ -) , 1,4-Butyl (-CH ₂ CH ₂ CH ₂ CH ₂ -) and similar groups.

The "C ₁ -C ₁₀ alkylene group" is a linear saturated alkyl group having the formula -(CH ₂ ) _1-10 -. Examples of C ₁ -C ₁₀ alkylene include methylene, ethylene, propylene, butylene, pentylene, hexylene, heptylene, octylene, nonylylene, and decylene.

"Alkenylene" refers to an unsaturated branch having 2-18 carbon atoms and having two monovalent group centers obtained by removing two hydrogen atoms from the same carbon atom or two different carbon atoms of the parent alkene Chain or linear or cyclic alkyl. Typical alkenylene groups include (but are not limited to): 1,2-ethylene (-CH=CH-).

"Alkynylene" refers to an unsaturation with two monovalent groups having 2-18 carbon atoms and having two monovalent group centers obtained by removing two hydrogen atoms from the same carbon atom or two different carbon atoms of the parent alkyne Branched or linear or cyclic alkyl. Typical alkynylene groups include, but are not limited to, acetylene (-C≡C-), propargyl (-CH ₂ C≡C-) and 4-pentynyl (-CH ₂ CH ₂ CH ₂ C≡C-).

"Aryl" refers to a carbocyclic aromatic group. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, and anthracenyl. The carbocyclic aromatic group or heterocyclic aromatic group may be unsubstituted or substituted with one or more groups including but not limited to the following: -C ₁ -C ₈ alkyl, -O-(C ₁ -C ₈ alkyl), -aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH ₂ , -C(O)NHR',- C(O)N(R') ₂ -NHC(O)R', -S(O) ₂ R', -S(O)R', -OH, -halogen, -N ₃ , -NH ₂ ,- NH(R'), -N(R') ₂ and -CN; wherein each R'is independently selected from H, -C ₁ -C ₈ alkyl and aryl.

The "C ₅ -C ₂₀ aryl group" is an aryl group having 5 to 20 carbon atoms in the carbocyclic aromatic ring. Examples of C ₅ -C ₂₀ aryl groups include, but are not limited to, phenyl, naphthyl, and anthracenyl. The C ₅ -C ₂₀ aryl group may be substituted or unsubstituted as described above for the aryl group. The "C ₅ -C ₁₄ aryl group" is an aryl group having 5 to 14 carbon atoms in the carbocyclic aromatic ring. Examples of C ₅ -C ₁₄ aryl groups include, but are not limited to, phenyl, naphthyl, and anthracenyl. The C ₅ -C ₁₄ aryl group may be substituted or unsubstituted as described above for the aryl group.

其中苯基可未經取代或經至多四個包括但不限於以下之基團取代：-C₁ -C₈ 烷基、-O-(C₁ -C₈ 烷基)、-芳基、-C(O)R'、-OC(O)R'、-C(O)OR'、-C(O)NH₂ 、-C(O)NHR'、-C(O)N(R')₂ -NHC(O)R'、-S(O)₂ R'、-S(O)R'、-OH、-鹵素、-N₃ 、-NH₂ 、-NH(R')、-N(R')₂ 及-CN；其中各R'係獨立地選自H、-C₁ -C₈ 烷基及芳基。"Aryl" is an aryl group that has two covalent bonds and can be in ortho, meta or para configuration as shown in the following structure:

The phenyl group may be unsubstituted or substituted with up to four groups including but not limited to the following: -C ₁ -C ₈ alkyl, -O-(C ₁ -C ₈ alkyl), -aryl, -C (O)R', -OC(O)R', -C(O)OR', -C(O)NH ₂ , -C(O)NHR', -C(O)N(R') _2- NHC(O)R', -S(O) ₂ R', -S(O)R', -OH, -halogen, -N ₃ , -NH ₂ , -NH(R'), -N(R' ) ₂ and -CN; wherein each R'is independently selected from H, -C ₁ -C ₈ alkyl and aryl.

"Arylalkyl" refers to an acyclic alkyl group in which a hydrogen atom bonded to a carbon atom (usually a terminal or sp ³ carbon atom) is replaced by an aryl group. Typical arylalkyl groups include, but are not limited to, benzyl, 2-phenylethyl-1-yl, 2-phenylvin-1-yl, naphthylmethyl, 2-naphthylethyl-1-yl, 2- Naphthylethen-1-yl, naphthobenzyl, 2-naphthophenyleth-1-yl and similar groups. The arylalkyl group contains 6 to 20 carbon atoms, for example, the alkyl portion (including alkyl, alkenyl, or alkynyl) of the arylalkyl group is 1 to 6 carbon atoms and the aryl portion is 5 to 14 carbon atoms .

"Heteroarylalkyl" refers to an acyclic alkyl group in which a hydrogen atom bonded to a carbon atom (usually a terminal or sp ³ carbon atom) is replaced by a heteroaryl group. Typical heteroarylalkyl groups include, but are not limited to, 2-benzimidazolylmethyl, 2-furylethyl and the like. Heteroarylalkyl contains 6 to 20 carbon atoms, for example, the alkyl portion (including alkyl, alkenyl or alkynyl) of heteroarylalkyl is 1 to 6 carbon atoms and the heteroaryl portion is 5 to 14 Carbon atoms and 1 to 3 heteroatoms selected from N, O, P and S. The heteroaryl portion of the heteroarylalkyl group can be a monocyclic ring with 3 to 7 ring members (2 to 6 carbon atoms) or 7 to 10 ring members (4 to 9 carbon atoms and 1 to 3 Bicyclic ring (heteroatom selected from N, O, P and S), for example: bicyclic [4,5], [5,5], [5,6] or [6,6] system.

"Substituted alkyl", "substituted aryl" and "substituted arylalkyl" respectively mean an alkyl, aryl and aryl group in which one or more hydrogen atoms are each independently replaced by a substituent alkyl. Typical substituents include (but are not limited to) -X, -R, -O ^- , -OR, -SR, -S ^- , -NR ₂ , -NR ₃ , =NR, -CX ₃ , -CN, -OCN, -SCN, -N=C=O, -NCS, -NO, -NO ₂ , =N ₂ , -N ₃ , NC(=O)R, -C(=O)R, -C(=O)NR _{^{2, -SO 3 -, -SO 3}} H, -S (= O) 2 R, -OS (= O) 2 OR, -S (= O) 2 NR, -S (= O) R, -OP ( =O)(OR) ₂ , -P(=O)(OR) ₂ , -PO ^- ₃ , -PO ₃ H ₂ , -C(=O)R, -C(=O)X, -C(= _{S) R, -CO 2 R,} -CO 2 -, -C (= S) OR, -C (= O) SR, -C (= S) SR, -C (= O) NR 2, -C ( =S)NR ₂ , -C(=NR)NR ₂ , wherein each X is independently halogen: F, Cl, Br or I; and each R is independently -H, C ₂ -C ₁₈ alkyl, C ₆ -C ₂₀ aryl, C ₃ -C ₁₄ heterocycle, protecting group or prodrug moiety. The alkylene, alkenylene and alkynylene groups described above may also be substituted similarly.

"Heteroaryl" and "heterocyclic ring" refer to a ring system in which one or more ring atoms are heteroatoms (such as nitrogen, oxygen, and sulfur). The heterocyclic group contains 3 to 20 carbon atoms and 1 to 3 heteroatoms selected from N, O, P, and S. The heterocyclic ring can be a monocyclic ring having 3 to 7 ring members (2 to 6 carbon atoms and 1 to 3 heteroatoms selected from N, O, P, and S) or having 7 to 10 ring members (4 to 9 carbon atoms and 1 to 3 heteroatoms selected from N, O, P and S) bicyclic ring, for example: bicyclic ring [4,5], [5,5], [5,6] or [6,6 ]system.

Exemplary heterocycles are described in, for example, Paquette, Leo A., "Principles of Modern Heterocyclic Chemistry" (WA Benjamin, New York, 1968), specifically Chapters 1, 3, 4, 6, 7 and 9; "The Chemistry of Heterocyclic Compounds, A series of Monographs" (John Wiley & Sons, New York, 1950), Volumes 13, 14, 16, 19 and 28 of the specific language; and J. Am. Chem. Soc. (1960) 82 : 5566.

Examples of heterocyclic rings include (for example and are not limited to) pyridyl, dihydropyridinyl, tetrahydropyridinyl (piperidinyl), thiazolyl, tetrahydrothienyl, tetrahydrothienyl oxysulfide, pyrimidinyl, furanyl, Thienyl, pyrrolyl, pyrazolyl, imidazolyl, tetrazolyl, benzofuranyl, thionaphthyl, indolyl, indolenyl, quinolinyl, isoquinolinyl, benzimidazolyl, piper Pyridinyl, 4-piperidinone, pyrrolidinyl, 2-pyrrolidinone, pyrrolinyl, tetrahydrofuranyl, bis-tetrahydrofuranyl, tetrahydropiperanyl, bis-tetrahydropiperanyl, tetrahydroquine Linyl, tetrahydroisoquinolinyl, decahydroquinolinyl, octahydroisoquinolinyl, azetyl, triazinyl, 6H-1,2,5-thiadiazinyl, 2H,6H-1, 5,2-Dithiazinyl, thienyl, thioanthranyl, piperanyl, isobenzofuranyl, oxacenyl, oxanyl, phenoxathiyl, 2H-pyrrolyl, isothiazolyl, iso Oxazolyl, pyrazinyl, pyridazinyl, indolazinyl, isoindolyl, 3H-indolyl, 1H-indazolyl, purinyl, 4H-quinazinyl, phthalazinyl, naphthyridinyl , Quinoxalinyl, quinazolinyl, octolinyl, pteridine, 4aH-carbazolyl, carbazolyl, β-carbolinyl, phenanthridinyl, acridinyl, pyrimidinyl, phenantholinyl, phenanthrene Azinyl, phenothiazinyl, furanyl, phenanthrazinyl, isopropyl, oxacyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperazinyl, indolinyl , Isoindolinyl, quindinyl, morpholinyl, oxazolidinyl, benzotriazolyl, benzisoxazolyl, oxindole, benzoxazolinyl and isatinyl .

For example and without limitation, carbon-bonded heterocycles are bonded at the following positions: 2, 3, 4, 5 or 6 of pyridine; 3, 4, 5 or 6 of pyridazine; 2 of pyrimidine 4, 5 or 6 position; 2, 3, 5 or 6 position of pyrazine; 2, 3, 4 or 5 position of furan, tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole; of oxazole, imidazole or thiazole Position 2, 4 or 5; Position 3, 4 or 5 of isoxazole, pyrazole or isothiazole; Position 2 or 3 of aziridine; Position 2, 3 or 4 of azetidine; Position 2 of quinoline , 3, 4, 5, 6, 7 or 8 positions; or 1, 3, 4, 5, 6, 7 or 8 positions of isoquinoline. More generally, carbon-bonded heterocycles include 2-pyridyl, 3-pyridyl, 4-pyridyl, 5-pyridyl, 6-pyridyl, 3-pyridazinyl, 4-pyridazinyl, 5-pyridyl Azinyl, 6-pyridazinyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl, 2-pyrazinyl, 3-pyrazinyl, 5-pyrazinyl, 6-pyrimidinyl Azinyl, 2-thiazolyl, 4-thiazolyl, or 5-thiazolyl.

For example and without limitation, the nitrogen-bonded heterocycle is bonded at the following positions: aziridine, azetidine, pyrrole, pyrrolidine, 2-pyrroline, 3-pyrroline, imidazole, imidazole 1 of pyridine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline, 1H-indazole Position; 2-position of isoindole or isoindoline; 4-position of morpholine; and 9-position of carbazole or β-carboline. More generally, the nitrogen-bonded heterocycle includes 1-aziridinyl, 1-azetidinyl, 1-pyrrolyl, 1-imidazolyl, 1-pyrazolyl, and 1-piperidinyl.

"C ₃ -C ₈ heterocyclic ring" refers to an aromatic or non-aromatic C ₃ -C ₈ carbocyclic ring in which one to four ring carbon atoms are independently replaced by heteroatoms from the group consisting of O, S and N . Representative examples of C ₃ -C ₈ heterocycles include, but are not limited to, benzofuranyl, benzothiophene, indolyl, benzopyrazolyl, coumarin, isoquinolinyl, pyrrolyl, thienyl, Furyl, thiazolyl, imidazolyl, pyrazolyl, triazolyl, quinolinyl, pyrimidinyl, pyridyl, pyridonyl, pyrazinyl, pyridazinyl, isothiazolyl, isoxazolyl and tetrazole base. The C ₃ -C ₈ heterocycle may be unsubstituted or substituted with up to seven groups including but not limited to the following: -C ₁ -C ₈ alkyl, -O-(C ₁ -C ₈ alkyl), -aryl基, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH ₂ , -C(O)NHR', -C(O)N(R ') ₂ -NHC(O)R', -S(O) ₂ R', -S(O)R', -OH, -halogen, -N ₃ , -NH ₂ , -NH(R'),- N(R') ₂ and -CN; wherein each R'is independently selected from H, -C ₁ -C ₈ alkyl and aryl.

"C ₃ -C ₈ heterocyclic group" refers to the C ₃ -C ₈ heterocyclic group defined above, in which one hydrogen atom of the heterocyclic group is replaced by a bond. The C ₃ -C ₈ heterocyclic group may be unsubstituted or substituted with up to six groups including but not limited to: -C ₁ -C ₈ alkyl, -O-(C ₁ -C ₈ alkyl),- Aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH ₂ , -C(O)NHR', -C(O)N( R') ₂ -NHC(O)R', -S(O) ₂ R', -S(O)R', -OH, -halogen, -N ₃ , -NH ₂ , -NH(R'), -N(R') ₂ and -CN; wherein each R'is independently selected from H, -C ₁ -C ₈ alkyl and aryl.

"C ₃ -C ₂₀ heterocyclic ring" refers to an aromatic or non-aromatic C ₃ -C ₈ carbocyclic ring in which one to four ring carbon atoms are independently replaced by heteroatoms from the group consisting of O, S and N . The C ₃ -C ₂₀ heterocyclic ring may be unsubstituted or substituted with up to seven groups including but not limited to: -C ₁ -C ₈ alkyl, -O-(C ₁ -C ₈ alkyl), -aryl基, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH ₂ , -C(O)NHR', -C(O)N(R ') ₂ -NHC(O)R', -S(O) ₂ R', -S(O)R', -OH, -halogen, -N ₃ , -NH ₂ , -NH(R'),- N(R') ₂ and -CN; wherein each R'is independently selected from H, -C ₁ -C ₈ alkyl and aryl.

"C ₃ -C ₂₀ heterocyclic group" refers to the C ₃ -C ₂₀ heterocyclic group defined above, in which one hydrogen atom of the heterocyclic group is replaced by a bond.

"Carbocyclic ring" means a saturated or unsaturated ring having 3 to 7 carbon atoms in the form of a monocyclic ring or having 7 to 12 carbon atoms in the form of a bicyclic ring. The monocyclic carbocyclic ring has 3 to 6 ring atoms, more usually 5 or 6 ring atoms. The bicyclic carbocyclic ring has, for example, 7 to 12 ring atoms arranged in a bicyclic ring [4,5], [5,5], [5,6] or [6,6] system or arranged in a bicyclic ring [5,6] or [ 6,6] 9 or 10 ring atoms of the system. Examples of monocyclic carbon rings include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclopentyl Hexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cycloheptyl and cyclooctyl.

"C ₃ -C ₈ carbocyclic ring" is a 3-membered, 4-membered, 5-membered, 6-membered, 7-membered or 8-membered saturated or unsaturated non-aromatic carbocyclic ring. Representative C ₃ -C ₈ carbocyclic rings include, but are not limited to, -cyclopropyl, -cyclobutyl, -cyclopentyl, -cyclopentadienyl, -cyclohexyl, -cyclohexenyl, -1,3- Cyclohexadienyl, -1,4-cyclohexadienyl, -cycloheptyl, -1,3-cycloheptadienyl, -1,3,5-cycloheptatrienyl, -cyclooctyl And-cyclooctadienyl. The C ₃ -C ₈ carbocyclic group may be unsubstituted or substituted by one or more groups including but not limited to the following: -C ₁ -C ₈ alkyl, -O-(C ₁ -C ₈ alkyl) , -Aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH ₂ , -C(O)NHR', -C(O) N(R') ₂ -NHC(O)R', -S(O) ₂ R', -S(O)R', -OH, -halogen, -N ₃ , -NH ₂ , -NH(R' ), -N(R') ₂ and -CN; wherein each R'is independently selected from H, -C ₁ -C ₈ alkyl and aryl.

"C ₃ -C ₈ carbocyclic group" refers to the C ₃ -C ₈ carbocyclic group defined above, in which one hydrogen atom of the carbocyclic group is replaced by a bond.

"Linker" refers to a chemical moiety that contains a covalent bond or a series of atoms that makes the antibody covalently attached to the drug moiety. In various embodiments, the linker includes a divalent group, such as alkyldiyl, aryldiyl, heteroaryldiyl, moieties such as -(CR ₂ ) _n O(CR ₂ ) _n -, alkyl Repeating units of oxy groups ( such as polyethyleneoxy, PEG, polymethyleneoxy) and repeating units of alkylamine groups ( such as polyethyleneamino, Jeffamine ^TM ); and diacid esters and acetone Amines include succinate, succinamide, diglycolate, malonate, and hexanamide. In various embodiments, the linker may include one or more amino acid residues, such as valine, phenylalanine, lysine, and homolysine.

The term "palm" refers to a molecule that has the non-overlapping nature of the mirror image partner, and the term "non-palm" refers to a molecule that can be superimposed on its mirror image partner.

The term "stereoisomers" refers to compounds that have the same chemical composition, but differ in the arrangement of atoms or groups in space.

"Diastereomers" refer to stereoisomers that have two or more palmar centers and the molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting point, boiling point, spectral properties, and reactivity. Mixtures of diastereomers can be separated under high-resolution analysis procedures such as electrophoresis and chromatography.

"Enantiomers" refers to two stereoisomers of a compound that are non-superimposable mirror images of each other.

The definitions and conventions of stereochemistry used herein generally follow SP Parker, ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., Stereochemistry of Organic Compounds ( 1994) John Wiley & Sons, New York. Many organic compounds exist in optically active forms, that is, they have the ability to rotate the plane of plane-polarized light. When describing optically active compounds, the prefixes D and L, or R and S are used to indicate the absolute configuration of the molecule around its palm center. The prefixes d and l or (+) and (-) are used to designate plane-polarized light by the rotation of the compound, where (-) or l means that the compound is levorotatory. Compounds with the prefix (+) or d are dextrorotatory. For a given chemical structure, these stereoisomers are the same except that they are mirror images of each other. Specific stereoisomers can also be referred to as enantiomers, and mixtures of such isomers are often referred to as enantiomers. A 50:50 mixture of enantiomers is called a racemic mixture or a racemate, which can exist when there is no stereoselection or stereospecificity in a chemical reaction or process. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers that lacks optical activity.

The "leaving group" refers to a functional group that can be substituted by another functional group. Certain leaving groups are well known in the art, and examples include, but are not limited to, halide ( e.g., chloro, bromo, iodo), methanesulfonyl (methylsulfonyl), p-toluenesulfonyl Group (toluenesulfonyl), trifluoromethanesulfonyl (trifluoromethanesulfonate) and trifluoromethanesulfonate.

The term "protecting group" refers to a substituent generally used to block or protect a specific functional group when reacting other functional groups on a compound. For example, an "amino protecting group" is a substituent attached to an amino functional group in a blocking or protecting compound. Suitable amino protecting groups include, but are not limited to, acetyl, trifluoroacetyl, tertiary butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ), and 9-tinylmethyleneoxycarbonyl ( Fmoc). For a general description of protecting groups and their use, see TW Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991 or later editions. III. Method

This article provides a method for treating B-cell proliferative diseases (such as diffuse large B-cell lymphoma (DLBCL), such as relapsed/refractory DLBCL) in an individual (human individual) in need thereof, including administering to the individual an effective amount of (a) An immunoconjugate comprising an antibody that binds to CD79b linked to a cytotoxic agent and (b) at least one additional therapeutic agent, wherein the treatment (eg, treatment regimen) prolongs the progression-free survival (PFS) of the individual. In some embodiments, treatment (e.g., treatment regimen) prolongs the overall survival (OS) of the individual. This article also provides a method for treating B-cell proliferative diseases (such as diffuse large B-cell lymphoma (DLBCL), such as relapsed/refractory DLBCL) in an individual, including administering to the individual an effective amount of (a) an immunoconjugate , Which comprises an antibody that binds to CD79b linked to a cytotoxic agent and (b) at least one additional therapeutic agent, wherein the treatment (eg, treatment regimen) prolongs the overall survival (OS) of the individual. In some embodiments, after treatment with the immunoconjugate and at least one additional therapeutic agent, the individual achieves complete remission (CR), for example, as described in further detail elsewhere herein. (Additional details about CR are provided below.) In some embodiments, the at least one additional therapeutic agent is a chemotherapeutic agent. In some embodiments, the at least one additional therapeutic agent is a cytotoxic agent.

Provided herein is a method for treating a B cell proliferative disease in an individual in need (human individual), comprising administering to the individual an effective amount of (a) an immunoconjugate comprising an anti-CD79b antibody linked to a cytotoxic agent (That is, the anti-CD79b immunoconjugate and (b) alkylating agent. In some embodiments, the alkylating agent is 4-[5-[bis(2-chloroethyl)amino]-1-methylbenzene Bimidazol-2-yl]butyric acid or a salt or solvate thereof. In some embodiments, the alkylating agent is bendamustine or a salt or solvate thereof. In some embodiments, bendamustine or Its salt or solvate is bendamustine-HCl. In some embodiments, the anti-CD79b immunoconjugate is huMA79bv28-MC-vc-PAB-MMAE. In some embodiments, the immunoconjugate is polatuzumab vedotin (CAS Registration number 1313206-42-6).

This article provides a method for treating B-cell proliferative diseases (such as DLBCL, such as relapsed/refractory DLBCL) in an individual in need (a human individual), including administering to the individual an effective amount of (a) comprising a cytotoxic agent Linked immunoconjugates of anti-CD79b antibodies (ie anti-CD79b immunoconjugates), (b) alkylating agents, and (c) anti-CD20 agents (e.g. anti-CD20 antibodies), wherein the treatment (e.g., treatment regimen) prolongs the individual The progression-free survival (PFS). In some embodiments, treatment (e.g., treatment regimen) prolongs the overall survival (OS) of the individual. Also provided herein is a method of treating a B-cell proliferative disorder (e.g. DLBCL, such as relapsed/refractory DLBCL) in an individual, comprising administering to the individual an effective amount of (a) an immunization comprising an anti-CD79b antibody linked to a cytotoxic agent Conjugates (ie anti-CD79b immunoconjugates), (b) alkylating agents, and (c) anti-CD20 agents (e.g. anti-CD20 antibodies), wherein the treatment (e.g., treatment regimen) prolongs the overall survival (OS) of the individual. In some embodiments, the individual achieves complete remission (CR) after treatment with immunoconjugates, alkylating agents, and anti-CD20 agents, for example, as described in further detail elsewhere herein. (Additional details about CR are provided below.) In some embodiments, the anti-CD20 agent is an anti-CD20 antibody. In some embodiments, the anti-CD20 antibody system rituximab. In some embodiments, the anti-CD20 antibody system is a humanized B-Ly1 antibody. In some embodiments, the humanized B-Ly1 antibody system atolizumab. In some embodiments, the anti-CD20 antibody system ofatumumab, ublituximab and/or iblituximab. In some embodiments, the alkylating agent is 4-[5-[bis(2-chloroethyl)amino]-1-methylbenzimidazol-2-yl]butanoic acid or a salt or solvate thereof. In some embodiments, the alkylating agent is bendamustine or a salt or solvate thereof. In some embodiments, bendamustine or a salt or solvate thereof is bendamustine-HCl. In some embodiments, the anti-CD79b immunoconjugate is huMA79bv28-MC-vc-PAB-MMAE. In some embodiments, the immunoconjugate is polatuzumab vedotin (CAS Registry Number 1313206-42-6).

In some embodiments, B-cell proliferative disorders such as lymphoma (eg, B-cell non-Hodgkin's lymphoma (NHL)) and lymphocytic leukemia. Such lymphomas and lymphocytic leukemias include, for example, a) follicular lymphoma, b) small non-dividing cell lymphoma/Burkitt’s lymphoma (including endemic Burkitt’s lymphoma, sporadic Burkitt’s lymphoma) Lymphoma and non-Burkitt’s lymphoma), c) marginal zone lymphoma (including extranodal marginal zone B-cell lymphoma (mucosa-associated lymphoid tissue lymphoma, MALT), nodal marginal zone B-cell lymphoma, splenic margin Regional lymphoma), d) mantle cell lymphoma (MCL), =) large cell lymphoma (including B-cell diffuse large cell lymphoma (DLCL), diffuse mixed cell lymphoma, immunoblastic lymphoma, original Primary mediastinal B-cell lymphoma, angiocentral lymphoma-pulmonary B-cell lymphoma), f) hairy cell leukemia, g) lymphocytic lymphoma, Waldenstrom’s macroglobulinemia, h) Acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL), B-cell young lymphocytic leukemia, i) plasma cell tumor, plasma cell myeloma, multiple myeloma , Plasmacytoma, and/or j) Hodgkin’s disease.

In some embodiments, the B cell proliferative disorder is cancer. In some embodiments, the B-cell proliferative disorder is lymphoma, non-Hodgkin’s lymphoma (NHL), aggressive NHL, relapsed aggressive NHL, relapsed and slow NHL, refractory NHL, refractory and slow NHL, chronic lymphoma Cellular leukemia (CLL), small lymphocytic lymphoma, leukemia, hairy cell leukemia (HCL), acute lymphocytic leukemia (ALL), or mantle cell lymphoma. In some embodiments, the B-cell proliferative disease is NHL, such as mild NHL and/or aggressive NHL. In some embodiments, the B-cell proliferative disease line and mild follicular lymphoma or diffuse large B-cell lymphoma (DLBCL).

The term "co-administration" or "co-administering" refers to the combination of an anti-CD79b immunoconjugate and at least one additional therapeutic agent (for example, an alkylating agent and an anti-CD20 agent) as two One (or more) separate formulations (or as a single formulation containing an anti-CD79b immunoconjugate and at least one additive) are administered. In the case of using separate formulations, the co-administration can be carried out simultaneously or sequentially in any order, wherein there is preferably a time period during which all active agents simultaneously exert their biological activities. The anti-CD79b immunoconjugate and at least another therapeutic agent ( e.g., alkylating agent and anti-CD20 agent) are co-administered simultaneously or sequentially. In some embodiments, when all the therapeutic agents are administered together sequentially, the dose is administered in two separate administrations on the same day, or one agent is administered on day 1, and the other agent is administered on day 2 to day 7. , Such as co-administration between the 2nd and the 4th day. In some embodiments, the term "sequentially" means within 7 days after the dose of the first component, for example within 4 days after the dose of the first component; the term "simultaneously" means at the same time. The term "co-administration" with regard to the maintenance dose of the anti-CD79b immunoconjugate and at least one additional therapeutic agent ( for example , alkylating agent and anti-CD20 agent) means that the maintenance dose can be co-administered at the same time, if the treatment cycle is for all The medication is appropriate, for example weekly. Or, for example, the anti-CD79b immunoconjugate is administered every one to three days, and at least one additional therapeutic agent ( e.g. , alkylating agent and anti-CD20 agent) is administered weekly. Alternatively, the maintenance doses are co-administered sequentially within a day or several days.

The anti-CD79b immunoconjugates and other therapeutic agents ( eg , alkylating agents and anti-CD20 agents) provided herein for use in any of the treatment methods described herein will be formulated, administered, and administered in a manner consistent with good medical practice Vote. In this case, the consideration factors include the specific condition being treated, the specific mammal being treated, the clinical condition of the individual patient, the cause of the condition, the delivery site of the agent, the method of administration, the schedule of administration and the medical history. Other factors known to the industry. The antibody is not necessary, but is formulated with one or more agents currently used to prevent or treat the disorder as appropriate.

The co-administration amount and co-administration time of the anti-CD79b immunoconjugate and other therapeutic agents will depend on the type (species, sex, age, weight, etc.) and conditions of the patient being treated and the disease or condition being treated severity. The anti-CD79b immunoconjugate and at least one additional therapeutic agent (e.g., alkylating agent and anti-CD20 agent) are suitably co-administered to the patient at one time or in a series of treatments, for example on the same day or on the second day.

In some embodiments, the dose of an anti-CD79b immunoconjugate (e.g., huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin) is about 1.4-5 mg/kg, 1.8-4 mg/kg, 1.8-3.2 mg/kg , And/or any of 1.8-2.4 mg/kg. In some embodiments of any of these methods, the dose of the anti-CD79 immunoconjugate is about 1.4, 1.8, 2.0, 2.2, 2.4, 2.8, 3.2, 3.6, 4.0, 4.4, and/or 4.8 mg/kg Any of them. In some embodiments, the dose of the anti-CD79b immunoconjugate is about 1.8 mg/kg. In some embodiments, the dose of the anti-CD79b immunoconjugate is about 2.4 mg/kg. In some embodiments, the dose of the anti-CD79b immunoconjugate is about 3.2 mg/kg. In some embodiments, the dose of the anti-CD79b immunoconjugate is about 3.6 mg/kg. In some embodiments of any of these methods, the anti-CD79b immunoconjugate is administered as q3wk. In some embodiments, the anti-CD79b immunoconjugate is administered by intravenous infusion. The doses administered by infusion are about 1 μg/m ² to about 10,000 μg/m ² per dose, usually once a week, for a total of one, two, three, or four doses. Alternatively, the dose range is about 1 μg/m ² to about 1000 μg/m 2, about 1 μg/m ² to about 800 μg/m 2, about 1 μg/m ² to about 600 μg/m 2, about 1 μg/m ² To about 400 μg/m2, about 10 μg/m ² to about 500 μg/m2, about 10 μg/m ² to about 300 μg/m2, about 10 μg/m ² to about 200 μg/m2, and about 1 μg /m ² to about 200 μg/m ² . The dose can be administered once a day, once a week, multiple times a week, but less than once a day, multiple times a month but less than once a day, multiple times a month but less than once a week, and once a month Or intermittently to relieve or alleviate the symptoms of the disease. The administration can be continued at any disclosed interval until the symptoms of the B-cell proliferative disease or tumor being treated are relieved. In the case where the remission or relief is prolonged by this continuous administration, the administration can be continued after symptom relief or relief is achieved.

In some embodiments, the dose of the anti-CD20 agent ( eg , anti-CD20 antibody) is about 300-1600 mg/m ² and/or 300-2000 mg. In some embodiments, the dose of anti-CD20 antibody is about 300, 375, 600, 1000, or 1250 mg/m ² and/or any of 300, 1000, or 2000 mg. In some embodiments, the anti-CD20 antibody system rituximab is administered at a dose of 375 mg/m ² . In some embodiments, the anti-CD20 antibody system atolizumab is administered at a dose of 1000 mg/m ² . In some embodiments, the anti-CD20 antibody is administered as q3w (ie, administered every 3 weeks). In some embodiments, the non-trehalosylated anti-CD20 antibody (preferably the non-trehalosylated humanized B-Ly1 antibody) can be dosed on days 1, 8, and 15 of the 3 to 6 week dosage cycle Is 800 to 1600 mg (800 to 1200 mg in one embodiment), and then at a dose of 400 to 1200 (in one embodiment, 800 to 1200 mg) on day 1 of up to nine 3 to 4 week dose cycles In some embodiments, the dosage is a flat dose of 1000 mg in a three-week dosing schedule, and there may be an additional 1000 mg flat dose cycle in the second week.

Exemplary administration regimens for combination therapy of anti-CD79b immunoconjugates (e.g., huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin) and other agents include, but are not limited to, administration at about 1.4-5 mg/kg q3w Anti-CD79 immunoconjugate (e.g. huMA79bv28-MC-vc-PAB-MMAE), plus 375 mg/m ² q3w rituximab, and day 1 and day 2 of the 21-day cycle ( e.g. , q3w first Day and day 2) 25-120 mg/m ² bendamustine ( for example , bendamustine-HCl). In some embodiments, the anti-CD79 immunoconjugate is administered at about any of 1.8 mg/kg, 2.0 mg/kg, 2.2 mg/kg, 2.4 mg/kg, 3.2 mg/kg, or 4.0 mg/kg. In some embodiments, the anti-CD79b immunoconjugate is administered at about 1.8 mg/kg. In some embodiments, the anti-CD79b immunoconjugate is administered at about 2.4 mg/kg. In some embodiments, bendamustine ( eg , bendamustine-HCl) is administered at about 90 mg/m ² .

Another exemplary dosing regimen for combination therapy of anti-CD79b immunoconjugates (e.g., huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin) and other agents includes, but is not limited to, administration at about 1.4-5 mg/kg q3w With anti-CD79 immunoconjugates (such as huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin), plus 1000 mg/m ² q3w atolizumab, and 21-day cycle on day 1 and day 2 ( For example, q3w on day 1 and day 2) was administered 25-120 mg/m ² bendamustine (for example, bendamustine-HCl). In some embodiments, the anti-CD79 immunoconjugate is administered at about any of 1.8 mg/kg, 2.0 mg/kg, 2.2 mg/kg, 2.4 mg/kg, 3.2 mg/kg, or 4.0 mg/kg. In some embodiments, the anti-CD79b immunoconjugate is administered at about 1.8 mg/kg. In some embodiments, the anti-CD79b immunoconjugate is administered at about 2.4 mg/kg. In some embodiments, bendamustine (eg, bendamustine-HCl) is administered at about 90 mg/m ² .

The immunoconjugates provided herein (and any other therapeutic agents, such as alkylating agents and anti-CD20 agents) used in any of the treatment methods described herein can be administered by any suitable means, including parenteral, intrapulmonary and intranasal , And, if required for local treatment, intralesional administration. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. Administration may be by any suitable route, for example by injection, such as intravenous or subcutaneous injection, depending in part on whether the administration is short-lived or long-term. A variety of dosing regimens are covered herein, including but not limited to single or multiple administrations over multiple time points, rapid administration, and pulse infusion.

, 其中Ab係抗CD79b抗體，其包含(i)包含SEQ ID NO：21之胺基酸序列之HVR-H1；(ii)包含SEQ ID NO：22之胺基酸序列之HVR-H2；(iii)包含SEQ ID NO：23之胺基酸序列之HVR-H3；(iv)包含SEQ ID NO：24之胺基酸序列之HVR-L1；(v)包含SEQ ID NO：25之胺基酸序列之HVR-L2；(vi)包含SEQ ID NO：26之胺基酸序列之HVR-L3，並且其中p為1至8；(b)烷化劑，及(c)抗CD20抗體，其中治療(例如治療方案)延長個體之無進展生存期(PFS)。在一些實施例中，治療(例如，治療方案)延長了個體之總生存期。本文還提供了在有此需要之個體(人類個體)中治療彌漫性大B細胞淋巴瘤(DLBCL)之方法，包括向個體投與有效量之：(a)包含下式之免疫結合物

, 其中Ab係抗CD79b抗體，其包含(i)包含SEQ ID NO：21之胺基酸序列之HVR-H1；(ii)包含SEQ ID NO：22之胺基酸序列之HVR-H2；(iii)包含SEQ ID NO：23之胺基酸序列之HVR-H3；(iv)包含SEQ ID NO：24之胺基酸序列之HVR-L1；(v)包含SEQ ID NO：25之胺基酸序列之HVR-L2；(vi)包含SEQ ID NO：26之胺基酸序列之HVR-L3，並且其中p為1至8；(b)烷化劑，及(c)抗CD20抗體，其中治療(例如治療方案)延長個體之總生存期(OS)。在一些實施例中，個體在用抗CD79b免疫結合物、烷化劑及抗CD20抗體治療後達成完全緩解(CR)。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：19之胺基酸序列之重鏈可變域(VH)及含有SEQ ID NO：20之胺基酸序列之輕鏈可變域(VL)。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：37之胺基酸序列之重鏈及含有SEQ ID NO：35之胺基酸序列之輕鏈。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：36之胺基酸序列之重鏈及含有SEQ ID NO：38之胺基酸序列之輕鏈。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：36之胺基酸序列之重鏈及含有SEQ ID NO：35之胺基酸序列之輕鏈。在一些實施例中，p為2至7，2至6，2至5，3至5，或3至4。在一些實施例中，p為3.4。在一些實施例中，抗CD79b免疫結合物係huMA79bv28-MC-vc-PAB-MMAE。在一些實施例中，免疫結合物係polatuzumab vedotin (CAS登記號1313206-42-6)。在一些實施例中，烷化劑係4-[5-[雙(2-氯乙基)胺基]-1-甲基苯并咪唑-2-基]丁酸或其鹽。在一些實施例中，烷化劑係苯達莫司汀或其鹽或溶劑化物。在一些實施例中，苯達莫司汀或其鹽或溶劑化物係苯達莫司汀-HCl。在一些實施例中，抗CD20抗體係利妥昔單抗、人源化B-Ly1抗體、阿托珠單抗、奧法木單抗、ublituximab或替伊莫單抗。This article provides a method for the treatment of diffuse large B-cell lymphoma (DLBCL) in an individual (human individual) in need thereof, which comprises administering to the individual an effective amount of: (a) an immunoconjugate of the following formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iii) ) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25 HVR-L2; (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and wherein p is 1 to 8; (b) alkylating agent, and (c) anti-CD20 antibody, wherein the treatment ( For example, treatment plan) to extend the progression-free survival (PFS) of the individual. In some embodiments, treatment (e.g., treatment regimen) prolongs the overall survival of the individual. This article also provides a method for treating diffuse large B-cell lymphoma (DLBCL) in an individual (human individual) in need thereof, comprising administering to the individual an effective amount of: (a) an immunoconjugate of the following formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iii) ) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25 HVR-L2; (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and wherein p is 1 to 8; (b) alkylating agent, and (c) anti-CD20 antibody, wherein the treatment ( For example, treatment plan) to extend the overall survival (OS) of the individual. In some embodiments, the individual achieves complete remission (CR) after treatment with an anti-CD79b immunoconjugate, alkylating agent, and anti-CD20 antibody. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain variable domain (VH) containing the amino acid sequence of SEQ ID NO: 19 and a light chain containing the amino acid sequence of SEQ ID NO: 20. Chain variable domain (VL). In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 37 and a light chain containing the amino acid sequence of SEQ ID NO: 35. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 36 and a light chain containing the amino acid sequence of SEQ ID NO: 38. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 36 and a light chain containing the amino acid sequence of SEQ ID NO: 35. In some embodiments, p is 2 to 7, 2 to 6, 2 to 5, 3 to 5, or 3 to 4. In some examples, p is 3.4. In some embodiments, the anti-CD79b immunoconjugate is huMA79bv28-MC-vc-PAB-MMAE. In some embodiments, the immunoconjugate is polatuzumab vedotin (CAS Registry Number 1313206-42-6). In some embodiments, the alkylating agent is 4-[5-[bis(2-chloroethyl)amino]-1-methylbenzimidazol-2-yl]butyric acid or a salt thereof. In some embodiments, the alkylating agent is bendamustine or a salt or solvate thereof. In some embodiments, bendamustine or a salt or solvate thereof is bendamustine-HCl. In some embodiments, the anti-CD20 antibody system rituximab, humanized B-Ly1 antibody, atolizumab, ofatumumab, ublituximab, or ibrituximab.

^* 21天週期^ǂ 週期2-6In some embodiments, the anti-CD79b immunoconjugate (eg, huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin) is administered at a dose of 1.8 mg/kg. Alternatively or additionally, in some embodiments, the alkylating agent (eg, bendamustine or bendamustine-HCl) is administered at a dose of 90 mg/m ² . Alternatively or additionally, in some embodiments, the anti-CD20 antibody is administered at a dose of 375 mg/m ² . In some embodiments, the anti-CD79b immunoconjugate, alkylating agent, and anti-CD20 antibody are administered for at least 6 21-day cycles. In some embodiments, for the 21-day cycle of cycle 1, the immunoconjugate is administered intravenously at a dose of 1.8 mg/kg on day 2 and at a dose of 90 mg/m2 on day 2 and day 3. The alkylating agent was administered internally, and the anti-CD20 antibody was administered intravenously at a dose of 375 mg/m2 on day 1, and wherein for cycles 2-6, in each 21-day cycle, 1.8 mg on day 1 The immunoconjugate was administered intravenously at a dose of /kg. The alkylating agent was administered intravenously at a dose of 90 mg/m2 on the first and second days, and intravenous at a dose of 375 mg/m2 on the first day Anti-CD20 antibody is administered. Exemplary dosing and administration schedules are provided in the following Table A1 : Table A1 : Exemplary dosing and administration schedules

^* 21 days cycle ^ǂ cycle 2-6

In some embodiments, the anti-CD79b immunoconjugate and the alkylating agent are administered sequentially on the second day of the first cycle. In some embodiments, the anti-CD79b immunoconjugate is administered before the alkylating agent. In some embodiments, the anti-CD79b immunoconjugate, alkylating agent, and anti-CD20 antibody are administered sequentially on day 1 of cycles 2-6. In some embodiments, the anti-CD20 antibody is administered before the anti-CD79b immunoconjugate, and the anti-CD79b immunoconjugate is administered before the alkylating agent. In some embodiments, the anti-CD79b immunoconjugate, alkylating agent and anti-CD20 antibody are further administered after the sixth cycle. In some embodiments, every cycle after the 6th cycle, every 21-day cycle, the immunoconjugate is administered intravenously at a dose of 1.8 mg/kg on the first day, and on the first and second days The alkylating agent was administered intravenously at a dose of 90 mg/m ² and the anti-CD20 antibody was administered intravenously at a dose of 375 mg/m ² on the first day. In some embodiments, the anti-CD20 antibody is administered before the anti-CD79b immunoconjugate, and the anti-CD79b immunoconjugate is administered before the alkylating agent.

^* 21天週期^ǂ 週期2-6In some embodiments, the anti-CD79b immunoconjugate (eg, huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin) is administered at a dose of 1.8 mg/kg. Alternatively or additionally, in some embodiments, the alkylating agent (eg, bendamustine or bendamustine-HCl) is administered at a dose of 90 mg/m ² . Alternatively or additionally, in some embodiments, the anti-CD20 antibody is administered at a dose of 375 mg/m ² . In some embodiments, the anti-CD79b immunoconjugate, alkylating agent, and anti-CD20 antibody are administered for at least 6 21-day cycles. In some embodiments, the anti-CD79b immunoconjugate, alkylating agent, and anti-CD20 antibody are administered for no more than 6 21-day cycles. In some embodiments, for each 21-day cycle of cycles 1-6, the immunoconjugate is administered intravenously at a dose of 1.8 mg/kg on day 1, and 90 mg/m on day 1 and day 2. ^The alkylating agent was administered intravenously at a dose of ² , and the anti-CD20 antibody was administered intravenously at a dose of 375 mg/m ² on the first day. In some embodiments, every 21-day cycle after the 6th cycle, the immunoconjugate is administered intravenously at a dose of 1.8 mg/kg on day 1 and 90 mg/m on day 1 and day 2. ^The alkylating agent was administered intravenously at a dose of ² , and anti-CD20 antibody was administered intravenously at a dose of 375 mg/m ² on the first day. Another exemplary dosing and dosing schedule is provided in Table A2 below: Table A2 : Exemplary dosing and dosing schedule

^* 21 days cycle ^ǂ cycle 2-6

In some embodiments, the anti-CD79b immunoconjugate (e.g., huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin), alkylating agent and anti-CD20 antibody are administered sequentially on the first day of each 21-day cycle (e.g. , Period 1-6 and/or period beyond period 6). In some embodiments, the anti-CD20 antibody is administered before the anti-CD79b immunoconjugate, and the anti-CD79b immunoconjugate is administered before the alkylating agent. In some embodiments, the anti-CD79b immunoconjugate, alkylating agent, and anti-CD20 antibody are administered in any order.

, 其中Ab係抗CD79b抗體，其包含(i)包含VH之重鏈，該VH包含SEQ ID NO：19之胺基酸序列，及(ii)包含VL之輕鏈，該VL包含SEQ ID NO：20之胺基酸序列，其中p為2至5，(b)苯達莫司汀或其鹽或溶劑化物，及(c)利妥昔單抗，其中免疫結合物以1.8 mg/kg之劑量投與，苯達莫司汀或其鹽或溶劑化物以90 mg/m² 之劑量投與，並且利妥昔單抗以375 mg/m² 之劑量投與，並且其中該治療延長個體之無進展生存期(PFS)及/或總生存期(OS)。在一些實施例中，p為2至4或3至4。在一些實施例中，p為3.5。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：36所示胺基酸序列之重鏈及含有SEQ ID NO：35所示胺基酸序列之輕鏈。在一些實施例中，抗CD79b免疫結合物係huMA79bv28-MC-vc-PAB-MMAE。在一些實施例中，免疫結合物係polatuzumab vedotin (CAS登記號1313206-42-6)。在一些實施例中，苯達莫司汀或其鹽或溶劑化物係苯達莫司汀-HCl。在一些實施例中，個體在用抗CD79b免疫結合物、苯達莫司汀或其鹽或溶劑化物(例如苯達莫司汀-HCl)及利妥昔單抗治療後達成完全緩解(CR)。A method for treating diffuse large B-cell lymphoma (DLBCL) in an individual (human individual) in need is also provided, which comprises administering to the individual an effective amount of (a) an immunoconjugate comprising the following formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) a heavy chain comprising VH, which VH comprises the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, which VL comprises SEQ ID NO: The amino acid sequence of 20, where p is 2 to 5, (b) bendamustine or its salt or solvate, and (c) rituximab, where the immunoconjugate is at a dose of 1.8 mg/kg Administration, bendamustine or its salt or solvate is administered at a dose of 90 mg/m ² , and rituximab is administered at a dose of 375 mg/m ² , and wherein the treatment prolongs the individual’s absence Progressive survival (PFS) and/or overall survival (OS). In some embodiments, p is 2 to 4 or 3 to 4. In some embodiments, p is 3.5. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain containing the amino acid sequence shown in SEQ ID NO: 36 and a light chain containing the amino acid sequence shown in SEQ ID NO: 35. In some embodiments, the anti-CD79b immunoconjugate is huMA79bv28-MC-vc-PAB-MMAE. In some embodiments, the immunoconjugate is polatuzumab vedotin (CAS Registry Number 1313206-42-6). In some embodiments, bendamustine or a salt or solvate thereof is bendamustine-HCl. In some embodiments, the individual achieves complete remission (CR) after treatment with an anti-CD79b immunoconjugate, bendamustine or a salt or solvate thereof (such as bendamustine-HCl), and rituximab .

Anti-CD79b immunoconjugates (e.g., huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin), alkylating agents (e.g. bendamustine or bendamustine-HCl) and anti-CD20 antibodies (e.g. rituximab) Monoclonal antibodies) can be administered by the same route of administration or different routes of administration. In some embodiments, the anti-CD79b immunoconjugate is administered by intravenous, intramuscular, subcutaneous, topical, oral, transdermal, intraperitoneal, intraocular, implantation, inhalation, intrathecal, intraventricular, or intranasal administration. versus. In some embodiments, the alkylating agent (such as bendamustine, such as bendamustine-HCl) is administered intravenously, intramuscularly, subcutaneously, topically, oral, transdermal, intraperitoneal, intraocular, or It is administered by implantation, inhalation, intrathecal, intraventricular, or intranasal administration. In some embodiments, the anti-CD20 antibody (e.g., rituximab) is administered intravenously, intramuscularly, subcutaneously, topically, oral, transdermal, intraperitoneal, intraocular, by implantation, inhalation, intrathecal, Intraventricular or intranasal administration. In some embodiments, the anti-CD79b immunoconjugates, alkylating agents (for example, bendamustine, for example, bendamustine-HCl), and anti-CD20 antibodies (for example, rituximab) are administered by intravenous infusion. ). An effective amount of anti-CD79b immunoconjugates, alkylating agents (such as bendamustine, such as bendamustine-HCl) and anti-CD20 antibodies (such as rituximab) can be administered for the prevention or treatment of diseases .

^* 21天週期^ǂ 週期2-6In some embodiments, an anti-CD79b immunoconjugate (e.g., huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin), bendamustine (e.g., bendamustine-HCl), and rituximab are administered Anti-at least 6 21-day cycles, of which for the 21-day cycle of the first cycle, the anti-CD79b immunoconjugate was administered intravenously at a dose of 1.8 mg/kg on the second day, and bendamustine (such as bendamustine) Tine-HCl) was administered intravenously at a dose of 90 mg/m ² on days 2 and 3, and rituximab was administered intravenously at a dose of 375 mg/m ² on day 1, and wherein In each 21-day cycle after Cycle 1, in each 21-day cycle, the anti-CD79b immunoconjugate was administered intravenously at a dose of 1.8 mg/kg on day 1. Bendamustine (such as bendamustine) -HCl) was administered intravenously at a dose of 90 mg/m ² on the first and second days, and rituximab was administered intravenously at a dose of 375 mg/m ² on the first day. Exemplary dosing and administration schedules are provided in Table B1 below: Table B1 : Exemplary dosing and administration schedules

^* 21 days cycle ^ǂ cycle 2-6

In some embodiments, the anti-CD79b immunoconjugate and bendamustine (e.g., bendamustine-HCl) are administered sequentially on day 2 of cycle 1. In some embodiments, the anti-CD79b immunoconjugate is administered before bendamustine (eg, bendamustine-HCl). In some embodiments, the immunoconjugate, bendamustine (for example, bendamustine-HCl), and rituximab are administered sequentially on day 1 of cycles 2-6. In some embodiments, rituximab is administered before the anti-CD79b immunoconjugate, and the anti-CD79b immunoconjugate is administered before bendamustine (eg, bendamustine-HCl). In some embodiments, the anti-CD79b immunoconjugate, bendamustine (for example, bendamustine-HCl) and rituximab are further administered after the sixth cycle. In some embodiments, in each cycle after the sixth cycle, in each 21-day cycle, the immunoconjugate is administered intravenously at a dose of 1.8 mg/kg on the first day, and bendamustine (such as benzene Damustine-HCl) was administered intravenously at a dose of 90 mg/m ^{2 on} the first and second days, and rituximab was administered intravenously at a dose of 375 mg/m ² on the first day . In some embodiments, rituximab is administered before the anti-CD79b immunoconjugate, and the anti-CD79b immunoconjugate is administered before bendamustine (eg, bendamustine-HCl).

In some embodiments, the anti-CD79b immunoconjugate (eg, huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin) is administered at a dose of 1.8 mg/kg. Alternatively or additionally, in some embodiments, bendamustine (eg, bendamustine-HCl) is administered at a dose of 90 mg/m ² . Alternatively or additionally, in some embodiments, rituximab at a dose of 375 mg / m ² of administration. In some embodiments, the anti-CD79b immunoconjugate, bendamustine (eg, bendamustine-HCl), and rituximab are administered for at least 6 21-day cycles. In some embodiments, the administration of anti-CD79b immunoconjugates, bendamustine (eg, bendamustine-HCl), and rituximab does not exceed 6 21-day cycles. In some embodiments, for each 21-day cycle of cycles 1-6, the immunoconjugate is administered intravenously at a dose of 1.8 mg/kg on day 1, and bendamustine (e.g., bendamustine -HCl) was administered intravenously at a dose of 90 mg/m ² on the first and second days, and anti-rituximab was administered intravenously at a dose of 375 mg/m ² on the first day. In some embodiments, every 21-day cycle after the 6th cycle, the immunoconjugate is administered intravenously at a dose of 1.8 mg/kg on the first day, and bendamustine (for example, bendamustine) -HCl) was administered intravenously at a dose of 90 mg/m ² on the first and second days, and rituximab was administered intravenously at a dose of 375 mg/m ² on the first day. Another exemplary dosing and dosing schedule is provided in Table B2 below: Table B2 : Exemplary dosing and dosing schedule

In some embodiments, anti-CD79b immunoconjugates (e.g., huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin), bendamustine (e.g., bendamustine-HCl) and rituximab It is administered sequentially on the first day of each 21-day cycle (for example, cycles 1-6 and/or cycles other than cycle 6). In some embodiments, rituximab is administered before the anti-CD79b immunoconjugate, and the anti-CD79b immunoconjugate is administered before bendamustine (eg, bendamustine-HCl). In some embodiments, the anti-CD79b immunoconjugate, bendamustine (eg, bendamustine-HCl), and rituximab are administered in any order.

In any of the foregoing embodiments (e.g., the dosing and administration regimens provided herein), the anti-CD20 antibody (e.g., rituximab) is administered between 6 to 8 cycles. In some embodiments, the anti-CD20 antibody is administered for more than 8 cycles. In any of the foregoing embodiments (e.g., the dosing and administration regimens provided herein), wherein the individual suffers from peripheral neuropathy (e.g., before starting treatment) or develops peripheral neuropathy (e.g., during treatment), provided herein The dose of the anti-CD79b immunoconjugate (for example, huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin) in any of the examples was reduced to 1.4 mg/kg. Additionally or alternatively, wherein the individual suffers from neutropenia (e.g., grade 3-4 neutropenia) or thrombocytopenia (e.g., grade 3-4 thrombocytopenia), for example, before starting treatment , Or develop into neutropenia (e.g., grade 3-4 neutropenia) or thrombocytopenia (e.g., grade 3-4 thrombocytopenia), for example, during treatment, alkylating agents ( For example, the dose of bendamustine, such as bendamustine-HCl) is reduced to about 70 mg/m ² or about 50 mg/m ² .

In some embodiments of any of the methods described herein, CR (Complete Response/Complete Remission) is a complete radiographic response. In some embodiments, the complete radiographic response is assessed by computer tomography (CT). In some embodiments, the complete radiographic response is characterized by the following criteria: (a) by CT measurement, all target nodes or node masses in the individual have returned to the longest diameter ≤ 1.5 cm, (b) among the individual Any previously unmeasured lesions have disappeared, (c) there are no extralymphatic sites of the disease in the individual, and (d) no organs are huge (ie, abnormally enlarged organs). In some embodiments, the complete radiographic response is further characterized by normal bone marrow morphology. Alternatively or additionally, in some embodiments, CR is a complete metabolic response (CMR). In some embodiments, CMR is assessed by F ¹⁸ -2-fluoro-2-deoxy-d-glucose positron emission tomography-computed tomography (FDG-PET/CT). In some embodiments, CMR is characterized by the following criteria: (a) According to the Deauville 5-point scale, the score is 1 (FDG uptake is not higher than background), 2 (uptake≤mediastinum) or 3 (uptake>mediastinum but≤ The liver) has or does not have residual mass, where if the disease is not FDG-affinity, the residual mass is allowed, and (b) there is no evidence of FDG-affinity lesions in the bone marrow. Further details on the clinical staging and response criteria of lymphoma such as DLBCL are provided in, for example, Van Heertum et al. (2017) Drug Des. Devel. Ther. 11: 1719-1728; Cheson et al. (2016) Blood. 128: 2489-2496 ; Cheson et al. (2014) J. Clin. Oncol. 32(27): 3059-3067; Barrington et al. (2017) J. Clin. Oncol. 32(27): 3048-3058; Gallamini et al. (2014) Haematologica . 99(6): 1107-1113; Barrinton et al. (2010) Eur. J. Nucl. Med. Mol. Imaging. 37(10): 1824-33; Moskwitz (2012) Hematology Am Soc. Hematol. Educ. Program 2012: 397-401; and Follows et al. (2014) Br. J. Haematology 166: 34-49. The progress of any of the treatment methods provided herein can be monitored by techniques known in the art.

In some embodiments, PFS is measured from the date when a complete or partial response to treatment (for example, a recorded complete or partial response to treatment) first occurred to the date of disease progression, recurrence, or death due to any cause . In some embodiments, PFS is measured based on PET-CT or CT only, from the date of the first treatment to the first occurrence of progression or recurrence or death caused by any cause. In some embodiments, complete response, partial response, progression and/or recurrence are assessed according to the modified Lugano criteria, such as by PET/CT or CT measurement (see, for example, Cheson et al. (2014) "Recommendations for initial evaluation, staging, and response assessment of hodgkin and non-hodgkin lymphoma: The Lugano Classification." J. Clin. Oncol. 32:3059-67). In some embodiments, PFS is measured as the time from the start of treatment ( for example , with anti-CD79 immunoconjugates (for example, huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin), alkylating agents ( for example , bendamole The time from the treatment of stine, such as bendamustine-HCl) and anti-CD20 antibody ( e.g. , rituximab) to death. In some embodiments, the PFS is median PFS. In some embodiments, an anti-CD79 immunoconjugate (e.g., huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin), an alkylating agent ( e.g. bendamustine, for example bendamustine-HCl) and Treatment with anti-CD20 antibodies ( e.g. , rituximab) extends the individual’s PFS to at least about 6, 6.1, 6.2, 6.3, 6.4, 6.5, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17 (including any value between these Range) or more than 17 months. In some embodiments, an anti-CD79 immunoconjugate (e.g., huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin), an alkylating agent (e.g. bendamustine, for example bendamustine-HCl) and Treatment with anti-CD20 antibodies (e.g., rituximab) prolongs the individual's PFS to at least about 7.6 months. In some embodiments, an anti-CD79 immunoconjugate (for example, huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin), an alkylating agent ( for example , bendamustine or bendamustine-HCl) and Treatment with anti-CD20 antibodies ( e.g. , rituximab) prolongs the individual's PFS to at least about 11.1 months. In some embodiments, the treatment (eg, treatment regimen) prolongs the individual's PFS by at least 0.5, 1, 1.5, 2, 2.5 compared to an individual with DLBCL (eg, an individual with DLBCL who is not receiving treatment) , 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5 (including any range between these values), or any of them more than 8.5 months. In some embodiments, compared with receiving an alkylating agent (e.g., bendamustine, e.g., bendamustine-HCl) and an anti-CD20 antibody (e.g., rituximab) without an anti-CD79 immunoconjugate (e.g., Compared with individuals with DLBCL treated by huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin, treatment (eg, treatment regimen) increases the individual’s PFS by at least about 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 (including any range between these values), or any one of more than 8.5 months. In some embodiments, compared with receiving an alkylating agent ( e.g., bendamustine, e.g., bendamustine-HCl) and an anti-CD20 antibody (e.g., rituximab) without an anti-CD79 immunoconjugate (e.g., Compared with individuals with DLBCL treated with huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin), the median PFS is prolonged to at least about 11.1 months (e.g., at least about 6, 6.5, 7, 7.5, 8, Any one of 8.5, 9, 9.5, 10, 10.5 or 11, including any range between these equivalent values), the hazard ratio (HR) is equal to or less than 0.36. In some embodiments, the median PFS with a 95% confidence interval is between 6.2 and 13.9 months. In some embodiments, compared with receiving an alkylating agent (e.g., bendamustine, e.g., bendamustine-HCl) and an anti-CD20 antibody (e.g., rituximab) without an anti-CD79 immunoconjugate (e.g., Compared with individuals with DLBCL treated with huMA79bv28-MC-vc-PAB-MMAE or polatuzumab vedotin), the median PFS is extended to at least about 7.6 months (e.g., about 4, 4.5, 5, 5, 5.5, 6, Any one of 6.5, 7, or 7.5, including any range between these equivalent values), the hazard ratio (HR) is equal to or less than 0.34. In some embodiments, the median PFS with a 95% confidence interval is between 6.0 and 17.0 months.

In some embodiments, OS is measured as the time from diagnosis to death. In some embodiments, overall survival (OS) is measured from the beginning of treatment (for example, with anti-CD79 immunoconjugates, alkylating agents (for example, bendamustine, such as bendamustine-HCl) and The time period from anti-CD20 antibody (e.g., rituximab) to death (e.g., from any cause). In some embodiments, treatment with anti-CD79 immunoconjugates, alkylating agents (such as bendamustine, such as bendamustine-HCl), and anti-CD20 antibodies (such as rituximab) treats the individual’s OS Extend to at least about 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5 (including any range between these values) or more than 12.5 Any one of the month. In some embodiments, treatment with anti-CD79 immunoconjugates, alkylating agents (such as bendamustine, such as bendamustine-HCl), and anti-CD20 antibodies (such as rituximab) treats the individual’s OS Extend to at least about 12.4 months. In some embodiments, the treatment (e.g., treatment regimen) prolongs the individual's OS by at least about 0.5, 1, 1.5, 2, compared to an individual with DLBCL (e.g., an individual with DLBCL who is not receiving treatment). Any of 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5 (including any range between these values) or more than 7.5 months. In some embodiments, compared with receiving an alkylating agent (e.g., bendamustine, e.g., bendamustine-HCl) and an anti-CD20 antibody (e.g., rituximab) without an anti-CD79 immunoconjugate (e.g., Compared with individuals with DLBCL treated with huMA79bv28-MC-VC-PAB-MMAE or polatuzumab vedotin, treatment (eg, treatment regimen) increases the individual’s OS by at least about 0.5, 1, 1.5, 2, 2.5, 3 , 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5 (including any range between these values), or any one of more than 7.5 months. In some embodiments, compared with receiving an alkylating agent ( e.g., bendamustine, e.g., bendamustine-HCl) and an anti-CD20 antibody ( e.g. , rituximab) without an anti-CD79 immunoconjugate (e.g., Compared with individuals with DLBCL treated by huMA79bv28-MC-VC-PAB-MMAE or polatuzumab vedotin), the median OS is prolonged to at least about 12.4 months (e.g., at least about 6, 6.5, 7, 7.5, 8, 8.5 , 9, 9.5, 10, 10.5, 11, 11.5 or 12, including the range between these values), the hazard ratio (HR) is equal to or less than 0.42. In some embodiments, the median OS with a 95% confidence interval is at least 9.0 months.

In some embodiments, individuals are adults. In some embodiments, the individual has activated B cell DLBCL (ABC DLBCL). In some embodiments, the individual has germinal center B cell-like DLBCL (GCB DLBCL). In some embodiments, the individual has DLBCL not specified (DLBCL-NOS). In some embodiments, the NanoString Research-Only Lymphoma Subtype Test (LST) assay is used to perform DLBCL classification by cell of origin (COO). In some embodiments, the individual has dual manifestation lymphoma (DEL). DEL is DLBCL characterized by the overexpression of MYC and BLC2. In some embodiments, it is determined by immunohistochemistry (IHC) that the individual has DEL. In some embodiments, BCL2 (124) and MYC (Y69) monoclonal antibodies are used for IHC assays. In some embodiments, IHC verification is performed on the Ventana Benchmark XT platform. In some embodiments, MYC overexpression is characterized by ≥ 40% of tumor cell nuclei as positive staining, and BCL2 overexpression is characterized by ≥ 50% of tumor cell nuclei as positive staining. In some embodiments, the individual has relapsed-refractory DLBCL (RR-DLBCL). In some embodiments, RR-DLBCL is characterized by (a) the appearance of new lesions or an increase in the size of previously involved disease sites after remission is reached by more than 50%, and (b) any previously identified abnormalities with a short axis greater than 1 cm The maximum diameter of a node, or the sum of product diameters (SPD) of an abnormal node, increased by more than 50%, (c) SPD of any previously determined abnormal node increased by more than 50% from the lowest point, and/or (d) during treatment New lesions appear during or at the end of treatment. Additional details on the criteria for characterizing RR-DLBCL are provided in Cheson et al. (1999) J. Clin. Oncol. 17(4): 1244. In some embodiments, the individual does not have grade 3 follicular lymphoma, transforming and mild non-Hodgkin's lymphoma, or central nervous system (CNS) lymphoma. In some embodiments, the individual is not suitable for autologous stem cell transplantation (ASCT), for example, first-line ASCT, second-line ASCT, third-line ASCT, or beyond third-line ASCT. In some embodiments, the individual has a large B-cell lymphoma rich in T cells/histioblasts. In some embodiments, the individual has high-grade B-cell lymphoma with MYC and BCL-2 and/or BCL-6 rearrangements. In some embodiments, the individual has unspecified (NOS) high-grade B-cell lymphoma. In some embodiments, the individual has primary mediastinal (thymic) large B-cell lymphoma. In some embodiments, the individual has Epstein-Barr virus positive DLBCL not specified (NOS). In some embodiments, the individual has at least one two-dimensional measurable lesion during the imaging scan, the longest dimension of which is> 1.5 cm. In some embodiments, the individual has not received prior therapy for DLBCL (e.g., prior chemotherapy or prior antibody therapy). In some embodiments, the individual has experienced at least one previous therapy for DLBCL. In some embodiments, the individual has experienced at least two previous therapies for DLBCL. In some embodiments, the individual has experienced at least three previous therapies for DLBCL. In some embodiments, the individual has experienced more than three previous therapies for DLBCL. In some embodiments, the individual's previous autologous stem cell transplantation (ASCT) has failed, such as relapse after ASCT or refractory to ASCT. In some embodiments, the individual has received prior treatment with an alkylating agent (e.g., bendamustine, such as bendamustine-HCl). In some embodiments, the duration of the previous treatment with an alkylating agent (eg, bendamustine, eg, bendamustine-HCl) is ≥ 1 year. In some embodiments, the individual has already received an anti-CD20 agent, such as a previous therapy with an anti-CD-20 antibody. In some embodiments, the individual is refractory to the most recent previous therapy. In some embodiments, the most recent prior therapy is the standard of care therapy. In some embodiments, if the individual exhibits a partial response, minimal response, or no response to the treatment, the individual is refractory to the treatment. In some embodiments, the individual system is female. In some embodiments, an adult with unspecified (NOS) DLBCL has received at least one previous therapy (eg, for DLBCL).

In some embodiments, DLBCL is BCL2 positive (e.g., for BCL2 gene rearrangement, t(14;18)(q32;q21) is positive). In some embodiments, DLBCL is BCL2 negative (for example, t(14;18)(q32;q21) is negative for BCL2 gene rearrangement). In some embodiments, the individual has (for example, further has) one or more of the following characteristics: (a) at least one two-dimensional measurable lesion during imaging scan, the longest dimension of which is defined as> 1.5 cm; (b) Life expectancy is at least 24 weeks; (c) Eastern Cooperative Oncology Group (ECOG) physical strength status is 0, 1 or 2; (d) Sufficient blood function.

In some embodiments, the individual does not have a history of severe allergies or allergic reactions to humanized or murine monoclonal antibodies (MAb or recombinant antibody-related fusion proteins); known sensitivities or allergic reactions to mouse products; Or bendamustine (such as bendamustine-HCl), rituximab or atolizumab contraindications. In some embodiments, the individual does not have a history of sensitivity to mannitol. In some embodiments, for purposes other than lymphoma symptom control, the individual does not receive corticosteroids at doses> 30 mg/day prednisone or equivalent.

In some embodiments of any of these methods, if the administration is intravenous, the initial infusion time of the anti-CD79b immunoconjugate or another therapeutic agent may be longer than the subsequent infusion time, for example, the initial infusion is about 90%. Minutes, and then infusion for approximately 30 minutes (if the initial infusion is well tolerated).

, 其中Ab係抗CD79b抗體，其包含(i)包含SEQ ID NO：21之胺基酸序列之HVR-H1；(ii)包含SEQ ID NO：22之胺基酸序列之HVR-H2；(iii)包含SEQ ID NO：23之胺基酸序列之HVR-H3；(iv)包含SEQ ID NO：24之胺基酸序列之HVR-L1；(v)包含SEQ ID NO：25之胺基酸序列之HVR-L2；(vi)包含SEQ ID NO：26之胺基酸序列之HVR-L3，並且其中p為1至8; (b)烷化劑，及(c)根據本文所述任何實施例之抗CD20抗體。在一些實施例中，改善PFS包括改善中位PFS。在一些實施例中，該方法改善了患有DLBCL之個體之OS。在一些實施例中，改善OS包括改善中位OS。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：19之胺基酸序列之重鏈可變域(VH)及含有SEQ ID NO：20之胺基酸序列之輕鏈可變域(VL)。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：37之胺基酸序列之重鏈及含有SEQ ID NO：35之胺基酸序列之輕鏈。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：36之胺基酸序列之重鏈及含有SEQ ID NO：38之胺基酸序列之輕鏈。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：36之胺基酸序列之重鏈及含有SEQ ID NO：35之胺基酸序列之輕鏈。在一些實施例中，p為2至7，2至6，2至5，3至5，或3至4。在一些實施例中，p為3.4。在一些實施例中，抗CD79b免疫結合物係huMA79bv28-MC-vc-PAB-MMAE。在一些實施例中，免疫結合物係polatuzumab vedotin (CAS登記號1313206-42-6)。在一些實施例中，烷化劑係4-[5-[雙(2-氯乙基)胺基]-1-甲基苯并咪唑-2-基]丁酸或其鹽。在一些實施例中，烷化劑係苯達莫司汀或其鹽或溶劑化物。在一些實施例中，苯達莫司汀或其鹽或溶劑化物係苯達莫司汀-HCl。在一些實施例中，抗CD20抗體係利妥昔單抗。在一些實施例中，PFS之改善係相對於用烷化劑(例如苯達莫司汀，例如苯達莫司汀-HCl)及抗CD20抗體而不用抗CD79免疫結合物(例如，huMA79bv28-MC-vc-PAB-MMAE或polatuzumab vedotin)治療的患有DLBCL之個體。在一些實施例中，OS之改善係相對於用烷化劑(例如苯達莫司汀，例如苯達莫司汀-HCl)及抗CD20抗體而不用抗CD79免疫結合物(例如，huMA79bv28-MC-vc-PAB-MMAE或polatuzumab vedotin)治療的患有DLBCL之個體。This article provides a method for improving the PFS of an individual (human individual) suffering from DLBCL, comprising administering to the individual an effective amount of: (a) an immunoconjugate comprising the following formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iii) ) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25 (Vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and wherein p is 1 to 8; (b) alkylating agent, and (c) according to any embodiment described herein The anti-CD20 antibody. In some embodiments, improving PFS includes improving median PFS. In some embodiments, the method improves OS in individuals with DLBCL. In some embodiments, improving OS includes improving median OS. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain variable domain (VH) containing the amino acid sequence of SEQ ID NO: 19 and a light chain containing the amino acid sequence of SEQ ID NO: 20. Chain variable domain (VL). In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 37 and a light chain containing the amino acid sequence of SEQ ID NO: 35. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 36 and a light chain containing the amino acid sequence of SEQ ID NO: 38. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 36 and a light chain containing the amino acid sequence of SEQ ID NO: 35. In some embodiments, p is 2 to 7, 2 to 6, 2 to 5, 3 to 5, or 3 to 4. In some examples, p is 3.4. In some embodiments, the anti-CD79b immunoconjugate is huMA79bv28-MC-vc-PAB-MMAE. In some embodiments, the immunoconjugate is polatuzumab vedotin (CAS Registry Number 1313206-42-6). In some embodiments, the alkylating agent is 4-[5-[bis(2-chloroethyl)amino]-1-methylbenzimidazol-2-yl]butyric acid or a salt thereof. In some embodiments, the alkylating agent is bendamustine or a salt or solvate thereof. In some embodiments, bendamustine or a salt or solvate thereof is bendamustine-HCl. In some embodiments, the anti-CD20 antibody system rituximab. In some embodiments, the improvement of PFS is relative to the use of alkylating agents (such as bendamustine, such as bendamustine-HCl) and anti-CD20 antibodies instead of anti-CD79 immunoconjugates (such as huMA79bv28-MC -vc-PAB-MMAE or polatuzumab vedotin) treated individuals with DLBCL. In some embodiments, the improvement of OS is relative to the use of alkylating agents (such as bendamustine, such as bendamustine-HCl) and anti-CD20 antibodies instead of anti-CD79 immunoconjugates (such as huMA79bv28-MC -vc-PAB-MMAE or polatuzumab vedotin) treated individuals with DLBCL.

, 其中Ab係抗CD79b抗體，其包含(i)包含SEQ ID NO：21之胺基酸序列之HVR-H1；(ii)包含SEQ ID NO：22之胺基酸序列之HVR-H2；(iii)包含SEQ ID NO：23之胺基酸序列之HVR-H3；(iv)包含SEQ ID NO：24之胺基酸序列之HVR-L1；(v)包含SEQ ID NO：25之胺基酸序列之HVR-L2；(vi)包含SEQ ID NO：26之胺基酸序列之HVR-L3，並且其中p為1至8; (b)烷化劑，及(c)根據本文所述任何實施例之抗CD20抗體。在一些實施例中，改善OS包括改善中位OS。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：19之胺基酸序列之重鏈可變域(VH)及含有SEQ ID NO：20之胺基酸序列之輕鏈可變域(VL)。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：37之胺基酸序列之重鏈及含有SEQ ID NO：35之胺基酸序列之輕鏈。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：36之胺基酸序列之重鏈及含有SEQ ID NO：38之胺基酸序列之輕鏈。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：36之胺基酸序列之重鏈及含有SEQ ID NO：35之胺基酸序列之輕鏈。在一些實施例中，p為2至7，2至6，2至5，3至5，或3至4。在一些實施例中，p為3.4。在一些實施例中，抗CD79b免疫結合物係huMA79bv28-MC-vc-PAB-MMAE。在一些實施例中，免疫結合物係polatuzumab vedotin (CAS登記號1313206-42-6)。在一些實施例中，烷化劑係4-[5-[雙(2-氯乙基)胺基]-1-甲基苯并咪唑-2-基]丁酸或其鹽。在一些實施例中，烷化劑係苯達莫司汀或其鹽或溶劑化物。在一些實施例中，苯達莫司汀或其鹽或溶劑化物係苯達莫司汀-HCl。在一些實施例中，抗CD20抗體係利妥昔單抗。在一些實施例中，OS之改善係相對於用烷化劑(例如苯達莫司汀，例如苯達莫司汀-HCl)及抗CD20抗體而不用抗CD79免疫結合物(例如，huMA79bv28-MC-vc-PAB-MMAE或polatuzumab vedotin)治療的患有DLBCL之個體。This article provides a method for improving the OS of an individual (human individual) suffering from DLBCL, comprising administering to the individual an effective amount of: (a) an immunoconjugate comprising the following formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iii) ) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25 (Vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and wherein p is 1 to 8; (b) alkylating agent, and (c) according to any embodiment described herein The anti-CD20 antibody. In some embodiments, improving OS includes improving median OS. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain variable domain (VH) containing the amino acid sequence of SEQ ID NO: 19 and a light chain containing the amino acid sequence of SEQ ID NO: 20. Chain variable domain (VL). In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 37 and a light chain containing the amino acid sequence of SEQ ID NO: 35. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 36 and a light chain containing the amino acid sequence of SEQ ID NO: 38. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 36 and a light chain containing the amino acid sequence of SEQ ID NO: 35. In some embodiments, p is 2 to 7, 2 to 6, 2 to 5, 3 to 5, or 3 to 4. In some examples, p is 3.4. In some embodiments, the anti-CD79b immunoconjugate is huMA79bv28-MC-vc-PAB-MMAE. In some embodiments, the immunoconjugate is polatuzumab vedotin (CAS Registry Number 1313206-42-6). In some embodiments, the alkylating agent is 4-[5-[bis(2-chloroethyl)amino]-1-methylbenzimidazol-2-yl]butyric acid or a salt thereof. In some embodiments, the alkylating agent is bendamustine or a salt or solvate thereof. In some embodiments, bendamustine or a salt or solvate thereof is bendamustine-HCl. In some embodiments, the anti-CD20 antibody system rituximab. In some embodiments, the improvement of OS is relative to the use of alkylating agents (such as bendamustine, such as bendamustine-HCl) and anti-CD20 antibodies instead of anti-CD79 immunoconjugates (such as huMA79bv28-MC -vc-PAB-MMAE or polatuzumab vedotin) treated individuals with DLBCL.

Also provided is the use of the anti-CD79b immunoconjugate described herein in the preparation or preparation of a medicament, which is combined with at least one additional therapeutic agent, for example, an alkylating agent (such as bendamustine, such as bendamustine- HCl) and an anti-CD20 antibody (for example, rituximab) are used in combination to treat DLBCL in an individual in need thereof (for example, a human individual with one or more characteristics as described above), wherein the administration of the prolonged individual PFS and/or OS.

, 其中Ab係抗CD79b抗體，其包含(i)包含SEQ ID NO：21之胺基酸序列之HVR-H1；(ii)包含SEQ ID NO：22之胺基酸序列之HVR-H2；(iii)包含SEQ ID NO：23之胺基酸序列之HVR-H3；(iv)包含SEQ ID NO：24之胺基酸序列之HVR-L1；(v)包含SEQ ID NO：25之胺基酸序列之HVR-L2；(vi)包含SEQ ID NO：26之胺基酸序列之HVR-L3，並且其中p在1及8之間，該抗體用於治療有需要之個體(人類個體)中之彌漫性大B細胞淋巴瘤(DLBCL)的方法，該方法包括向個體投與有效量之免疫結合物、烷化劑及抗C20抗體，其中該治療延長個體之無進展生存期(PFS)及/或總生存期(OS)。在一些實施例中，免疫結合物用於本文描述之方法。在一些實施例中，免疫結合物包含抗CD79b抗體，其包含(i)包含VH之重鏈，該VH包含SEQ ID NO：19之胺基酸序列，及(ii)包含VL之輕鏈，該VL包含SEQ ID NO：20之胺基酸序列。Provide immunoconjugates containing the following formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iii) ) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25 HVR-L2; (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and wherein p is between 1 and 8. This antibody is used for the treatment of diffuse in individuals in need (human individuals) A method for large B-cell lymphoma (DLBCL), the method comprising administering to an individual an effective amount of an immunoconjugate, an alkylating agent, and an anti-C20 antibody, wherein the treatment extends the individual’s progression-free survival (PFS) and/or Overall survival (OS). In some embodiments, immunoconjugates are used in the methods described herein. In some embodiments, the immunoconjugate comprises an anti-CD79b antibody, which comprises (i) a heavy chain comprising VH, the VH comprising the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, the VL contains the amino acid sequence of SEQ ID NO:20.

, 其中Ab係抗CD79b抗體，其包含(i)包含VH之重鏈，該VH包含SEQ ID NO：19之胺基酸序列，及(ii)包含VL之輕鏈，該VL包含SEQ ID NO：20之胺基酸序列，其中p在2與5之間，該抗體用於治療有需要之個體(人類個體)中之彌漫性大B細胞淋巴瘤(DLBCL)的方法，該方法包括投與個體有效量之(a)免疫結合物，(b)苯達莫司汀(例如苯達莫司汀-HCl)及(c)利妥昔單抗之量，其中免疫結合物以1.8 mg/kg之劑量投與，苯達莫司汀(例如苯達莫司汀-HCl)係以90 mg/m² 之劑量投與，並且利妥昔單抗以375 mg/m² 之劑量投與，並且其中該治療延長個體之無進展生存期(PFS)及/或總生存期(OS)。在一些實施例中，免疫結合物用於根據本文描述之方法的用途中。在一些實施例中，p介於3與4之間。在一些實施例中，p為3.5。在一些實施例中，免疫結合物包含含有重鏈之抗CD79b抗體，該重鏈包含SEQ ID NO：36之胺基酸序列，並且其中該輕鏈包含SEQ ID NO：35之胺基酸序列。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：37之胺基酸序列之重鏈及含有SEQ ID NO：35之胺基酸序列之輕鏈。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：36之胺基酸序列之重鏈及含有SEQ ID NO：38之胺基酸序列之輕鏈。 IV. 包含抗 CD79b 抗體及藥物 / 細胞毒性劑之免疫結合物 ( 「抗 CD79b 免疫結合物」 ) Also provided is an immunoconjugate comprising the formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) a heavy chain comprising VH, which VH comprises the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, which VL comprises SEQ ID NO: The amino acid sequence of 20, where p is between 2 and 5. The antibody is used in a method for treating diffuse large B-cell lymphoma (DLBCL) in an individual in need (human individual), the method comprising administration to the individual The effective amount of (a) immunoconjugate, (b) bendamustine (such as bendamustine-HCl) and (c) rituximab, wherein the immunoconjugate is at 1.8 mg/kg Dosage administration, bendamustine (for example, bendamustine-HCl) is administered at a dose of 90 mg/m ² , and rituximab is administered at a dose of 375 mg/m ² , and wherein The treatment prolongs the progression-free survival (PFS) and/or overall survival (OS) of the individual. In some embodiments, the immunoconjugate is used in use according to the methods described herein. In some embodiments, p is between 3 and 4. In some embodiments, p is 3.5. In some embodiments, the immunoconjugate comprises an anti-CD79b antibody comprising a heavy chain, the heavy chain comprising the amino acid sequence of SEQ ID NO: 36, and wherein the light chain comprises the amino acid sequence of SEQ ID NO: 35. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 37 and a light chain containing the amino acid sequence of SEQ ID NO: 35. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 36 and a light chain containing the amino acid sequence of SEQ ID NO: 38. IV. Immunoconjugates containing anti- CD79b antibodies and drugs / cytotoxic agents ( "anti- CD79b immunoconjugates" )

In some embodiments, the anti-CD79b immunoconjugate includes an anti-CD79b antibody (Ab) that targets cancer cells (eg, diffuse large B-cell lymphoma (DLBCL) cells), a drug moiety (D), and an Ab attached to D Connect the subsection (L). In some embodiments, the anti-CD79b antibody is linked to the linker portion (L) via one or more amino acid residues (such as lysine and/or cysteine). In some formulas Ab-(LD)p, wherein: (a) the Ab line binds to the CD79b anti-CD79b antibody on the surface of cancer cells (such as DLBCL cells); (b) the L line linker; (c) the D line cytotoxicity Agent; (d) The range of p is 1-8.

Exemplary anti-CD79b immunoconjugates include Formula I: (I) Ab-(LD) _p where p is 1 to about 20 ( e.g. , 1 to 15, 1 to 10, 1 to 8, 2 to 5, or 3 to 4) . In some embodiments, the number of drug moieties that can bind to an anti-CD79b antibody is limited by the number of free cysteine residues. In some embodiments, free cysteine residues are introduced into the antibody amino acid sequence by methods described elsewhere herein. Exemplary anti-CD79b immunoconjugates of Formula I include, but are not limited to, anti-CD79b antibodies containing 1, 2, 3, or 4 engineered cysteine amino acids (Lyon, R. et al. (2012) Methods in Enzym. 502:123-138). In some embodiments, one or more free cysteine residues are already present in the anti-CD79b antibody without the use of engineering. In this case, the existing free cysteine residues can be used to make the anti-CD79b antibody Combined with drugs/cytotoxic agents. In some embodiments, the anti-CD79b antibody is exposed to reducing conditions, and then the antibody is allowed to bind to the drug/cytotoxic agent to produce one or more free cysteine residues. A. Exemplary linkers

"Linker" (L) is a bifunctional or multifunctional moiety that can be used to link one or more drug moieties (D) to an anti-CD79b antibody (Ab) to form an anti-CD79b immunoconjugate of Formula I. In some embodiments, linkers with reactive functional groups can be used to prepare anti-CD79b immunoconjugates, and these functional groups are used for covalent attachment to drugs and anti-CD79b antibodies. For example, in some embodiments, the cysteine thiol of an anti-CD79b antibody (Ab) can form a bond with a linker or a reactive functional group of a drug-linker intermediate to prepare an anti-CD79b immunoconjugate.

In one aspect, the linker has a functional group capable of reacting with free cysteine present on the anti-CD79b antibody to form a covalent bond. Exemplary reactive functional groups include, but are not limited to, for example, maleimide, haloacetamide, α-haloacetamide, activated esters (such as succinimide ester, 4-nitrophenyl ester, Pentafluorophenyl ester, tetrafluorophenyl ester), acid anhydride, acid chloride, sulfonyl chloride, isocyanate and isothiocyanate. See, for example, Klussman et al. (2004), Bioconjugate Chemistry 15(4): 765-773, page 766 of the binding method and the examples herein.

In some embodiments, the linker has a functional group capable of reacting with an electrophilic group present on the anti-CD79b antibody. Exemplary electrophilic groups include, but are not limited to, for example, aldehyde and ketone carbonyl groups. In some embodiments, the heteroatom of the reactive functional group of the linker can react with the electrophilic group on the antibody and form a covalent bond with the antibody unit. Exemplary reactive functional groups include, but are not limited to, for example, hydrazine, oxime, amine, hydrazine, thiosemicarbazide, hydrazine carboxylate, and arylhydrazine.

In some embodiments, the linker includes one or more linker components. Exemplary linker components include, for example, 6-maleiminohexyl ("MC"), maleiminopropionyl ("MP"), valine-citrulline Acid ("val-cit" or "vc"), alanine-phenylalanine ("ala-phe"), p-aminobenzyloxycarbonyl ("PAB"), N-butanedioxin 4 -(2-Pyridylthio)pentanoate ("SPP") and 4-(N-maleiminomethyl)cyclohexane-1-carboxylate ("MCC"). Various linker components are known in the art, some of which are described below.

In some embodiments, the linker is a "cleavable linker" to promote drug release. Non-limiting exemplary cleavable linkers include acid labile linkers ( e.g. containing hydrazone), protease-sensitive ( e.g. peptidase sensitive) linkers, photolabile linkers or disulfide-containing linkers (Chari et al. Human, Cancer Research 52:127-131 (1992); US 5208020).

其中A為「延伸子單元」，且a為整數0至1；W爲「胺基酸單元」，且w為整數0至12；Y為「間隔子單元」，且y為0、1或2；且Ab、D及p係如上文對於式I所定義。此等連接子之示範性實施例描述於美國專利第7,498,298號中，該專利以引用的方式明確併入本文中。In certain embodiments, the linker (L) has the following formula II: (II)

Where A is an "extension subunit", and a is an integer from 0 to 1; W is an "amino acid unit" and w is an integer from 0 to 12; Y is a "spacer unit", and y is 0, 1 or 2 ; And Ab, D and p are as defined for Formula I above. Exemplary embodiments of these linkers are described in US Patent No. 7,498,298, which is expressly incorporated herein by reference.

In some embodiments, the linker component includes an "extension subunit" that connects the antibody to another linker component or drug moiety. The following shows a non-limiting exemplary extension subunit (where the wavy line indicates the site of covalent attachment to the antibody, drug or other linker component):

In some embodiments, the linker component includes "amino acid units." In some of these embodiments, the amino acid unit allows the linker to be cleaved by proteases, thereby helping to release the drug/cytotoxic agent from the anti-CD79b immunoconjugate when exposed to intracellular proteases (such as lysosomal enzymes) (Doronina et al. (2003) Nat. Biotechnol. 21:778-784). Exemplary amino acid units include, but are not limited to, dipeptides, tripeptides, tetrapeptides, and pentapeptides. Exemplary dipeptides include but are not limited to valine-citrulline (vc or val-cit), alanine-phenylalanine (af or ala-phe); phenylalanine-lysine (fk or phe-lys); Phenylalanine-homolysine (phe-homolys); and N-methyl-valine-citrulline (Me-val-cit). Exemplary tripeptides include, but are not limited to, glycine-valine-citrulline (gly-val-cit) and glycine-glycine-glycine (gly-gly-gly). The amino acid unit may comprise naturally occurring amino acid residues and/or minor amino acids and/or non-naturally occurring amino acid analogs, such as citrulline. Amino acid units can be designed and optimized for enzymatic cleavage by specific enzymes, such as tumor-associated proteases, cathepsins B, C and D, or plasmin proteases.

In some embodiments, the linker component includes a "spacer" unit that connects the antibody to the drug moiety either directly or via an extender unit and/or an amino acid unit. The spacer unit can be either "self-decomposing type" or "non-self-decomposing type". The "non-self-decomposing" spacer unit is a spacer unit where part or all of the spacer unit remains bound to the drug part after ADC lysis. Examples of non-self-decomposing spacer units include, but are not limited to, glycine spacer units and glycine-glycine spacer units. In some embodiments, the enzymatic cleavage of the ADC containing the glycine-glycine spacer unit by the tumor cell-related protease results in the release of the glycine-glycine-drug moiety from the rest of the ADC. In some of these embodiments, the glycine-glycine-drug moiety undergoes a hydrolysis step in the tumor cell, thereby cleaving the glycine-glycine spacer unit from the drug moiety.

其中Q為-C₁ -C₈ 烷基、-O-(C₁ -C₈ 烷基)、-鹵素、-硝基或-氰基；m為在0至4之範圍內之整數；且p在1至約20之範圍內。在一些實施例中，p在1至10、1至7、1至5、或1至4之範圍內。The "self-decomposing" spacer unit allows the release of the drug portion. In certain embodiments, the spacer unit of the linker comprises a p-aminobenzyl unit. In some of these embodiments, the p-aminobenzyl alcohol is connected to the amino acid unit via an amide bond, and a carbamate, methyl carbamate or carbonate is formed between the benzyl alcohol and the drug (Hamann et al. People (2005) Expert Opin. Ther. Patents (2005) 15:1087-1103). In some embodiments, the spacer unit is p-aminobenzyloxycarbonyl (PAB). In some embodiments, the anti-CD79b immunoconjugate includes a self-decomposing linker, which includes the following structure:

Wherein Q is -C ₁ -C ₈ alkyl, -O-(C ₁ -C ₈ alkyl), -halogen, -nitro or -cyano; m is an integer in the range of 0 to 4; and p In the range of 1 to about 20. In some embodiments, p is in the range of 1 to 10, 1 to 7, 1 to 5, or 1 to 4.

Other examples of self-decomposing spacers include, but are not limited to, aromatic compounds that are electronically similar to the PAB group, such as 2-aminoimidazole-5-methanol derivatives (US Patent No. 7,375,078; Hay et al. (1999) Bioorg Med. Chem. Lett. 9:2237) and o- or p-aminobenzyl acetal. In some embodiments, spacers that undergo cyclization upon hydrolysis of the amide bond can be used, such as substituted and unsubstituted 4-aminobutyric acid amides (Rodrigues et al. (1995) Chemistry Biology 2:223) , Appropriately substituted bicyclic [2.2.1] and bicyclic [2.2.2] ring systems (Storm et al. (1972) J. Amer. Chem. Soc. 94:5815) and 2-aminophenyl propionate (Amsberry et al. (1990) J. Org. Chem. 55:5867). The linkage of drugs to the α-carbon of glycine residues is another example of self-decomposing spacers that can be used in ADCs (Kingsbury et al. (1984) J. Med. Chem. 27:1447).

In some embodiments, the linker L may be a dendritic linker for covalently linking more than one drug moiety to an antibody via a branched multifunctional linker moiety (Sun et al. (2002) Bioorganic & Medicinal Chemistry Letters 12: 2213 -2215; Sun et al. (2003) Bioorganic & Medicinal Chemistry 11:1761-1768). The dendritic linker can increase the molar ratio of drug to antibody, that is, the loading capacity, which is related to the performance of ADC. Therefore, when the antibody carries only one reactive cysteine thiol group, numerous drug moieties can be connected via a dendritic linker.

val-cit (IV)

MC-val-cit (V)

MC-val-cit-PAB 其中(Ab)係抗CD79b抗體，(D)係藥物/細胞毒性劑，「Val-Cit」係纈胺酸 - 瓜胺酸二肽，MC係6-順丁烯二醯亞胺基己醯基，PAB係對胺基苯甲基氧基羰基，p係1到約20 (例如，1至15、1至10、1至8、2至5，或3至4)。The following non-limiting exemplary linkers are shown in the case of anti-CD79 immunoconjugates of formula III, IV, and V: (III)

val-cit (IV)

MC-val-cit (V)

MC-val-cit-PAB (Ab) is an anti-CD79b antibody, (D) is a drug/cytotoxic agent, "Val-Cit" is a valine-citrulline dipeptide, and MC is a 6-male Aminohexyl, PAB is a p-aminobenzyloxycarbonyl, p is 1 to about 20 (for example, 1 to 15, 1 to 10, 1 to 8, 2 to 5, or 3 to 4) .

, (VII)

, (VIII)

, (IX)

, (X)

, 其中X為：

Y為：

; 各R獨立地為H或C₁ -C₆ 烷基；且n為1至12。In some embodiments, the anti-CD79b immunoconjugate comprises the structure of any one of the following formula VI-V: (VI)

, (VII)

, (VIII)

, (IX)

, (X)

, Where X is:

Y is:

; Each R is independently H or C ₁ -C ₆ alkyl; and n is 1-12.

Generally, peptide linkers can be prepared by forming peptide bonds between two or more amino acids and/or peptide fragments. For example, these peptide bonds can be prepared according to a liquid phase synthesis method (for example, E. Schröder and K. Lübke (1965) "The Peptides", Vol. 1, pp. 76-136, Academic Press).

In some embodiments, the linker is substituted with a group that adjusts solubility and/or reactivity. As a non-limiting example, such as a sulfonate group (-SO ₃ ^-) or ammonium group of the charged substituent may increase water soluble linker reagent of the reagent and facilitate the linker and / or a coupling reaction with the antibody drug moiety , Or contribute to the coupling reaction of Ab-L (anti-CD79b antibody-linker intermediate) and D, or DL (drug/cytotoxic agent-linker intermediate) and Ab, which can be used to prepare anti-CD79b immunoconjugates It depends on the synthesis route. In some embodiments, part of the linker is coupled to the antibody and part of the linker is coupled to the drug, and then the anti-CD79 Ab-(linker portion) ^{a is} coupled to the drug/cytotoxic agent-(linker portion) ^b to The anti-CD79b immunoconjugate of formula I is formed. In some of these embodiments, the anti-CD79b antibody contains more than one (linker portion) ^a substituent to allow more than one drug/cytotoxic agent to couple to the anti-CD79b antibody in the anti-CD79b immunoconjugate of Formula I.

The anti-CD79b immunoconjugates provided herein clearly cover (but are not limited to) anti-CD79b immunoconjugates prepared with the following linker reagents: bis-maleimino-trioxyethylene glycol (BMPEO), N -(β-Maleimidinyl propyloxy)-N-hydroxysuccinimidyl ester (BMPS), N-(ε-maleiminohexyloxy) Succinonimide (EMCS), N-[γ-maleiminobutyryloxy] butanediimidate (GMBS), 1,6-hexane-bis-vinyl chloride ( HBVS), butanediimino 4-(N-maleiminomethyl)cyclohexane-1-carboxy-(6-aminohexanoate) (LC-SMCC), Maleimido benzyl-N-hydroxysuccinimide (MBS), 4-(4-N-maleiminophenyl) hydrazine butyrate (MPBH ), 3-(bromoacetamido) propionate succinimidyl ester (SBAP), iodoacetate succinimidyl ester (SIA), (4-iodoacetamido) butylaminobenzoate Diamido ester (SIAB), N-butanediamido-3-(2-pyridyldisulfide) propionate (SPDP), N-butanediamido-4-(2 -Pyridylthio) valerate (SPP), 4-(N-maleiminomethyl)cyclohexane-1-carboxylate succinimidyl ester (SMCC), 4-( P-Maleimidinyl phenyl)butyric acid succinimido ester (SMPB), 6-[(β-maleiminopropionylamino)hexanoic acid] succinic acid Imino ester (SMPH), iminothiocyclopentane (IT), sulfonic acid-EMCS, sulfonic acid-GMBS, sulfonic acid-KMUS, sulfonic acid-MBS, sulfonic acid-SIAB, Sulfonic acid group-SMCC and sulfonic acid group-SMPB and succinimide group-(4-vinylsulfonium) benzoate (SVSB), and include bis-maleimide reagent: disulfide group Dimaleiminoethane (DTME), 1,4-bismaleiminobutane (BMB), 1,4-bismaleimino-2, 3-Dihydroxybutane (BMDB), bismaleiminohexane (BMH), bismaleiminoethane (BMOE), BM(PEG) ₂ (shown below) and BM(PEG) ₃ (shown below); difunctional derivatives of imino esters (such as dimethyl diimide adipate hydrochloride), active esters (such as dibutylene suberate) Esters), aldehydes (such as glutaraldehyde), bis-azido compounds (such as bis(p-azidobenzyl) hexanediamine), bis-diazo derivatives (such as bis-(p-diazo Benzyl)-ethylenediamine), diisocyanates (such as toluene 2,6-diisocyanate), and dual-reactive fluorine compounds (such as 1,5-difluoro-2,4-dinitrobenzene). In some embodiments, the bis-maleimide reagent allows the thiol group of cysteine in the antibody to be linked to a thiol-containing drug moiety, linker, or linker-drug intermediate. Other functional groups that can react with thiol groups include, but are not limited to, iodoacetamide, bromoacetamide, vinyl pyridine, disulfide, pyridyl disulfide, isocyanate, and isothiocyanate.

Some useful linker reagents can be obtained from various commercial sources, such as Pierce Biotechnology (Rockford, IL), Molecular Biosciences (Boulder, CO), or synthesized according to the method described in this technology; for example, in Toki et al. (2002) J. Org. Chem. 67:1866-1872; Dubowchik et al. (1997) Tetrahedron Letters , 38:5257-60; Walker, MA (1995) J. Org. Chem. 60:5352-5355; Frisch et al. Human (1996) Bioconjugate Chem. 7:180-186; US 6214345; WO 02/088172; US 2003130189; US2003096743; WO 03/026577; WO 03/043583; and WO 04/032828.

Carbon 14-labeled 1-isothiocyanobenzyl-3-methyldiethylenetriaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for binding radionucleotides to antibodies. See, for example, WO94/11026. B. Anti- CD79b antibody

In some embodiments, the immunoconjugate (e.g., anti-CD79b immunoconjugate) comprises an anti-CD79b antibody comprising at least one, two, three, four, five or six HVRs selected from: (a ) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (c) HVR- comprising the amino acid sequence of SEQ ID NO: 23 H3; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (e) HVR-L2 comprising the amino acid of SEQ ID NO: 25; and (f) comprising the amino acid sequence of SEQ ID NO : HVR-L3 of 26. In some of these embodiments, the immunoconjugate comprises an anti-CD79 antibody comprising at least one of the following: (i) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23, and/or (ii) comprising an amine The base acid sequence of SEQ ID NO: 24 is HVR-L1. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody comprising at least one of the following: (i) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23, and/or (ii) comprising the amino acid HVR-L1 of SEQ ID NO: 24. In some embodiments, the immunoconjugate comprises an anti-CD79b antibody, the antibody comprising at least one, at least two, or all three VH HVR sequences selected from: (a) HVR comprising the amino acid sequence of SEQ ID NO: 21 -H1; (b) HVR-H2 comprising the amino acid sequence SEQ ID NO: 22; and (c) HVR-H3 comprising the amino acid sequence SEQ ID NO: 23. In some embodiments, the immunoconjugate comprises an anti-CD79b antibody comprising HVR-H3 containing the amino acid sequence of SEQ ID NO: 23. In another embodiment, the immunoconjugate comprises an anti-CD79b antibody comprising HVR-H3 containing the amino acid sequence of SEQ ID NO: 23 and HVR-L3 containing the amino acid of SEQ ID NO: 26. In some embodiments, the immunoconjugate comprises an anti-CD79b antibody comprising HVR-H3 containing the amino acid sequence of SEQ ID NO: 23, HVR-L3 containing the amino acid sequence of SEQ ID NO: 26, and containing amine The base acid sequence is HVR-H2 of SEQ ID NO: 22. In some embodiments, the immunoconjugate comprises an anti-CD79b antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21, (b) HVR comprising the amino acid sequence of SEQ ID NO: 22 -H2, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23.

In some embodiments, the immunoconjugate comprises an anti-CD79b antibody, the antibody comprising at least one, at least two, or all three VL HVR sequences selected from: (a) HVR comprising the amino acid sequence of SEQ ID NO: 24 -L1; (b) HVR-L2 comprising the amino acid sequence SEQ ID NO: 25; and (c) HVR-L3 comprising the amino acid sequence SEQ ID NO: 26. In some embodiments, the immunoconjugate comprises an anti-CD79b antibody, the antibody comprising at least one, at least two, or all three VL HVR sequences selected from: (a) HVR comprising the amino acid sequence of SEQ ID NO: 24 -L1; (b) HVR-L2 comprising the amino acid sequence SEQ ID NO: 25; and (c) HVR-L3 comprising the amino acid sequence SEQ ID NO: 26. In some embodiments, the immunoconjugate comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (c) Contains HVR-L3 of the amino acid sequence SEQ ID NO: 26. In some embodiments, the immunoconjugate comprises an anti-CD79b antibody, which comprises HVR-L1 containing the amino acid sequence of SEQ ID NO:24. In some embodiments, the immunoconjugate comprises an anti-CD79b antibody, which comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (b) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25 HVR-L2; (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:26.

In some embodiments, the immunoconjugate comprises an anti-CD79b antibody, which comprises (a) a VH domain, which comprises at least one, at least two, or all three VH HVR sequences selected from the group consisting of: (i) comprising an amino acid sequence HVR-H1 of SEQ ID NO: 21, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; And (b) a VL domain comprising at least one, at least two or all three VL HVR sequences selected from: (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24, (ii) comprising an amine HVR-L2 of the base acid sequence of SEQ ID NO: 25, and (iii) HVR-L3 of the amino acid sequence of SEQ ID NO: 26. In some embodiments, the immunoconjugate comprises an anti-CD79b antibody, the antibody comprising at least one of the following: (i) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23, and/or (ii) comprising the amino acid HVR-L1 of SEQ ID NO: 24.

In some embodiments, the immunoconjugate comprises an anti-CD79b antibody comprising (a) HVR-H1 comprising the amino acid sequence SEQ ID NO: 21; (b) HVR-H1 comprising the amino acid sequence SEQ ID NO: 22 H2; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (e) comprising the amino acid sequence of SEQ ID NO: HVR-L2 of 25; and (f) HVR-L3 of amino acid sequence SEQ ID NO: 26. In some embodiments, the immunoconjugate comprises at least one of the following: HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23, and/or HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24. In some embodiments, the immunoconjugate comprises an anti-CD79b antibody comprising (a) HVR-H1 comprising the amino acid sequence SEQ ID NO: 21; (b) HVR-H1 comprising the amino acid sequence SEQ ID NO: 22 H2; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (e) comprising the amino acid sequence of SEQ ID NO: HVR-L2 of 25; and (f) HVR-L3 of amino acid sequence SEQ ID NO: 26.

In some embodiments, the anti-CD79b immunoconjugate comprises a humanized anti-CD79b antibody. In some embodiments, the anti-CD79b antibody comprises HVR as in any of the embodiments provided herein, and further comprises a human acceptor framework, such as a human immunoglobulin framework or a human common framework. In some embodiments, the human acceptor framework is the human VL κ 1 (VL _KI ) framework and/or the VH framework VH _III . In some embodiments, the humanized anti-CD79b antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (c ) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (e) HVR- comprising the amino acid sequence of SEQ ID NO: 25 L2; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26. In some embodiments, the humanized anti-CD79b antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (c ) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (e) HVR- comprising the amino acid sequence of SEQ ID NO: 25 L2; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26.

In some embodiments, the immunoconjugate ( e.g., anti-CD79b immunoconjugate) comprises an anti-CD79 antibody comprising at least 90%, 91%, 92%, 93%, 94% with the amino acid sequence SEQ ID NO: 19 , 95%, 96%, 97%, 98%, 99% or 100% sequence identity of the heavy chain variable domain (VH) sequence. In certain embodiments, relative to the reference sequence, the amino acid sequence SEQ ID NO: 19 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% Or a 99% identical VH sequence contains substitutions ( such as conservative substitutions), insertions or deletions, but the anti-CD79b immunoconjugate containing that sequence retains the ability to bind to CD79b. In certain embodiments, in SEQ ID NO: 19, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted. In certain embodiments, in SEQ ID NO: 19, a total of 1 to 5 amino acids have been substituted, inserted, and/or deleted. In certain embodiments, the substitution, insertion, or deletion occurs in a region outside the HVR (ie, in the FR). In some embodiments, the immunoconjugate ( eg, anti-CD79b immunoconjugate) comprises the VH sequence of SEQ ID NO: 19, including post-translational modifications of that sequence. In some embodiments, the VH comprises one, two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21, (b) comprising the amino acid sequence of SEQ ID NO HVR-H2: 22, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 17 or SEQ ID NO: 23.

In some embodiments, the immunoconjugate ( e.g., anti-CD79b immunoconjugate) comprises an anti-CD79b antibody comprising at least 90%, 91%, 92%, 93%, 94% and the amino acid sequence SEQ ID NO: 20 %, 95%, 96%, 97%, 98%, 99% or 100% sequence identity of the light chain variable domain (VL). In certain embodiments, relative to the reference sequence, the amino acid sequence SEQ ID NO: 20 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% Or 99% identical VL sequences contain substitutions ( eg conservative substitutions), insertions or deletions, but the anti-CD79b immunoconjugates containing that sequence retain the ability to bind to CD79b. In certain embodiments, in SEQ ID NO: 20, a total of 1 to 10 amino acids have been substituted, inserted and/or deleted. In certain embodiments, in SEQ ID NO: 20, a total of 1 to 5 amino acids have been substituted, inserted and/or deleted. In certain embodiments, the substitution, insertion, or deletion occurs in a region outside the HVR (ie, in the FR). In some embodiments, the anti-CD79b immunoconjugate comprises an anti-CD79b antibody containing the VL sequence of SEQ ID NO: 20, including post-translational modifications of that sequence. In some embodiments, the VL comprises one, two or three HVRs selected from: (a) HVR-L1 comprising the amino acid sequence SEQ ID NO: 24; (b) comprising the amino acid sequence SEQ ID NO HVR-L2 of :25; and (c) HVR-L3 of SEQ ID NO:26 comprising the amino acid sequence. In some embodiments, the VL comprises one, two or three HVRs selected from: (a) HVR-L1 comprising the amino acid sequence SEQ ID NO: 24; (b) comprising the amino acid sequence SEQ ID NO HVR-L2 of :25; and (c) HVR-L3 of SEQ ID NO:26 comprising the amino acid sequence.

In some embodiments, the immunoconjugate ( eg , anti-CD79b immunoconjugate) comprises an anti-CD79b antibody, which comprises VH as in any embodiment provided herein, and VL as in any embodiment provided herein. In some embodiments, the immunoconjugate comprises an anti-CD79b antibody, which comprises the VH sequence and VL sequence in SEQ ID NO: 19 and SEQ ID NO: 20, respectively, including post-translational modifications of these sequences.

In some embodiments, the immunoconjugate ( eg , anti-CD79b immunoconjugate) comprises an anti-CD79b antibody that binds to the same epitope as the anti-CD79b antibody described herein. For example, in some embodiments, the immunoconjugate ( eg , anti-CD79b immunoconjugate) comprises an anti-CD79b antibody, which binds to an anti-CD79b antibody comprising the VH sequence of SEQ ID NO: 19 and the VL sequence of SEQ ID NO: 20 Same epitope.

In some embodiments, the immunoconjugate comprises an anti-CD79b antibody, which is a monoclonal antibody, a chimeric antibody, a humanized antibody, or a human antibody. In some embodiments, the immunoconjugate comprises an antigen-binding fragment of the anti-CD79b antibody described herein, such as Fv, Fab, Fab', scFv, miniature diabody, or F(ab') ₂ fragment. In some embodiments, the immunoconjugate comprises a substantially full-length anti-CD79b antibody, such as an IgG1 antibody or other antibody class or isotype as described elsewhere herein.

In some embodiments, the immunoconjugate comprises an anti-CD79b antibody, which comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 36, and wherein the light chain comprises the amino acid sequence of SEQ ID NO: 35. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 37 and a light chain containing the amino acid sequence of SEQ ID NO: 35. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 36 and a light chain containing the amino acid of SEQ ID NO: 38. C. Drugs / cytotoxic agents

The anti-CD79 immunoconjugate comprises an anti-CD79b antibody ( such as the anti-CD79b antibody described herein) that is conjugated to one or more drugs/cytotoxic agents, such as chemotherapeutics or drugs, growth inhibitors, Toxins ( for example, protein toxins, enzymatically active toxins of bacterial, fungal, plant or animal origin, or fragments thereof) or radioisotopes (i.e. radioactive conjugates). These immunoconjugates are targeted chemotherapeutic molecules that combine the properties of antibodies and cytotoxic drugs by targeted delivery of effective cytotoxic drugs to cancer cells expressing antigens (such as tumor cells) (Teicher, BA (2009) Current Cancer Drug Targets 9:982-1004), thereby enhancing the therapeutic index by maximizing efficacy and minimizing off-target toxicity (Carter, PJ and Senter PD (2008) The Cancer Jour. 14(3):154- 169; Chari, RV (2008) Acc. Chem. Res. 41:98-107. That is, the anti-CD79 conjugate selectively delivers an effective dose of the drug to cancer cells/tissues, thereby achieving higher selectivity ( That is, a lower effective dose), while increasing the therapeutic index ("treatment window") (Polakis P. (2005) Current Opinion in Pharmacology 5:382-387).

The anti-CD79 immunoconjugates used in the methods provided herein include anti-CD79 immunoconjugates with anticancer activity. In some embodiments, the anti-CD79 immunoconjugate includes an anti-CD79b antibody bound (ie, covalently linked) to a drug moiety. In some embodiments, the anti-CD79b antibody is covalently linked to the drug moiety via a linker. The drug part (D) of the anti-CD79 immunoconjugate may include any compound, part or group that has cytotoxic or cytostatic effects. The drug moiety can impart cytotoxicity and cytostatic effects through mechanisms including but not limited to tubulin binding, DNA binding or insertion, and inhibition of RNA polymerase, protein synthesis, and/or topoisomerase. Exemplary drugs include, but are not limited to, maytansinoid, dolastatin, aurisstatin, calicheamicin, anthracycline, duocarmycin, vinca alkaloid , Taxane, trichothecene, CC1065, camptothecin, elinafide and its stereoisomers, isosteres and analogs with cytotoxic activity And derivatives. (i) Maytansine and maytansinoids

In some embodiments, the anti-CD79b immunoconjugate comprises an anti-CD79b antibody bound to one or more maytansinoid molecules. Maytansinoids are derivatives of maytansines and are mitotic inhibitors that act by inhibiting tubulin polymerization. Maytansine was originally isolated from the East African shrub Maytenus serrata (US Patent No. 3896111). Subsequently, it was discovered that certain microorganisms also produce maytansinoids, such as maytansinol and C-3 maytansinol esters (US Patent No. 4,151,042). Synthetic maytansin is disclosed in, for example, U.S. Patent No. 4,137,230; No. 4,248,870; No. 4,256,746; No. 4,260,608; No. 4,265,814; No. 4,294,757; No. 4,307,016; No. 4,308,268; No. 4,308,269; No. 4,309,428; No. 4,313,946; No. 4,315,929; No. 4,317,821; No. 4,322,348; No. 4,331,598; No. 4,361,650; No. 4,364,866; No. 4,424,219; No. 4,450,254; No. 4,362,663; and No. 4,371,533.

The maytansinoid drug moiety is an attractive drug moiety in the antibody-drug conjugate because it is (i) relatively easy to prepare by fermentation or chemical modification or derivatization of fermentation products, and (ii) easily applicable Derivatization of functional groups bound to antibodies via non-disulfide linkers, (iii) stable in plasma, and (iv) effective against a variety of tumor cell lines.

Certain maytansinoids suitable for use as part of maytansinoids are known in this technology and can be isolated from natural sources according to known methods or produced using genetic engineering techniques ( see, for example, Yu et al. (2002) PNAS 99:7968-7973). Maytansin can also be prepared synthetically according to known methods.

Exemplary maytansinoids drug moieties include, but are not limited to, maytansinoids drug moieties with modified aromatic rings such as: C-19-dechlorination (U.S. Patent No. 4,256,746) (e.g., by reduction with lithium aluminum hydride) Ansamytocin (ansamytocin P2); C-20-hydroxy (or C-20-desmethyl) +/-C-19-dechlorination (US Patent Nos. 4,361,650 and 4,307,016) (for example, by by the use of Streptomyces (of Streptomyces) or actinomycetes (actinomyces), or demethylation be prepared using LAH to chlorine); and C-20- demethoxy, C-20- acyl group (-OCOR), +/- Dechlorination (US Patent No. 4,294,757) (for example, prepared by acylation using acetonitrile) and those with modifications in other positions of the aromatic ring.

Exemplary maytansinoid drugs also include those with modifications such as: C-9-SH (US Patent No. 4424219) (for example, prepared by reacting maytansinol with H ₂ S or P ₂ S ₅ ) ; C-14-alkoxymethyl (demethoxy/CH ₂ OR) (US 4331598); C-14-hydroxymethyl or oxymethyl (CH ₂ OH or CH ₂ OAc) (US patent No. 4450254) (for example, prepared from Nocardia); C-15-hydroxy/oxyl (US 4364866) (for example, prepared by transforming maytansinol with Streptomyces); C-15-methoxy (U.S. Patent Nos. 4313946 and 4315929) (e.g. isolated from Trewia nudlflora); C-18-N-desmethyl (U.S. Patent Nos. 4362663 and 4322348) (e.g. by using chain Maytansinol is prepared by demethylation by molds); and 4,5-deoxy (US 4371533) (for example, prepared by reducing maytansinol with titanium trichloride/LAH).

Many positions on the maytansin compound are suitable as bonding positions. For example, an ester bond can be formed by reacting with a hydroxyl group using conventional coupling techniques. In some embodiments, the reaction may occur at the C-3 position with a hydroxyl group, the C-14 position with a hydroxymethyl group, the C-15 position with a hydroxyl group, and the C-20 position with a hydroxyl group. In some embodiments, the linkage is formed at the C-3 position of maytansinol or a maytansinol analogue.

其中波形線指示美登木素藥物部分之硫原子與抗CD79b免疫結合物之連接子的共價連接。各R可獨立地為H或C₁ -C₆ 烷基。將醯胺基團連接到硫原子上之伸烷基鏈可以係甲烷基、乙烷基或丙基，即m係1、2或3 (US 633410；US 5208020；Chari等人 (1992)Cancer Res. 52:127-131；Liu等人(1996)Proc. Natl. Acad. Sci USA 93:8618-8623)。The maytansin drug part includes those with the following structure:

The wavy line indicates the covalent connection between the sulfur atom of the maytansinoid drug moiety and the linker of the anti-CD79b immunoconjugate. Each R may independently be H or C ₁ -C ₆ alkyl. The alkylene chain linking the amide group to the sulfur atom may be a methyl group, an ethylene group or a propyl group, ie m is 1, 2 or 3 (US 633410; US 5208020; Chari et al. (1992) Cancer Res . 52: 127-131; Liu et al. (1996) Proc Natl Acad Sci USA 93:... 8618-8623).

All stereoisomers of the maytansinoid drug portion are expected to be used in the anti-CD79b immunoconjugates provided herein, that is, any combination of R and S configurations at the opposing carbon (US 7276497; US 6913748; US 6441163 ; US 633410 (RE39151); US 5208020; Widdison et al., (2006) J. Med. Chem. 49:4392-4408, which is incorporated herein by reference in its entirety). In some embodiments, the maytansinoid drug moiety has the following stereochemistry:

其中波形線指示藥物之硫原子與抗CD79b免疫結合物之連接子(L)的共價連接。Exemplary embodiments of the drug portion of maytansinoid include but are not limited to DM1; DM3; and DM4 having the following structures:

The wavy line indicates the covalent connection between the sulfur atom of the drug and the linker (L) of the anti-CD79b immunoconjugate.

Ab -SPP-DM1

Ab-SMCC-DM1Other exemplary maytansinoid anti-CD79b immunoconjugates have the following structures and abbreviations (where Ab is an anti-CD79b antibody and p is 1 to about 20. In some embodiments, p is 1 to 10, and p is 1 to 7, p is 1 to 5, or p is 1 to 4):

Ab -SPP-DM1

Ab-SMCC-DM1

其中Ab為抗CD79b抗體；n為0、1或2；且p為1至約20。在一些實施例中，p為1至10，p為1至7，p為1至5，或p為1至4。An exemplary antibody-drug conjugate in which DM1 is connected to the thiol group of an antibody via a BMPEO linker has the following structure and abbreviations:

Where Ab is an anti-CD79b antibody; n is 0, 1, or 2; and p is 1 to about 20. In some embodiments, p is 1 to 10, p is 1 to 7, p is 1 to 5, or p is 1 to 4.

The immunoconjugates containing maytansinoids, their preparation methods and their therapeutic uses are disclosed in, for example, U.S. Patent Nos. 5,208,020 and 5,416,064; US 2005/0276812 A1; and European Patent EP 0 425 235 B1. The way of reference is expressly incorporated into this article. See also Liu et al., Proc. Natl. Acad. Sci. USA 93:8618-8623 (1996); and Chari et al., Cancer Research 52:127-131 (1992).

In some embodiments, the anti-CD79b antibody-maytansinoid conjugate can be prepared by chemically linking the anti-CD79b antibody to the maytansinoid molecule without significantly impairing the biological activity of the antibody or maytansinoid molecule. See, for example, U.S. Patent No. 5,208,020 (the disclosure of which is expressly incorporated herein by reference). In some embodiments, anti-CD79b immunoconjugates with an average of 3-4 maytansinoid molecules bound to each antibody molecule have been shown to be effective in enhancing the cytotoxicity of target cells without negatively affecting the function or solubility of the antibody . In some cases, it is expected that even one toxin/antibody molecule will increase cytotoxicity over the use of naked anti-CD79b antibodies.

Exemplary linking groups for the preparation of antibody-maytansin conjugates include, for example, those described herein and U.S. Patent No. 5,208,020; EP Patent 0 425 235 B1; Chari et al. Cancer Research 52:127- 131 (1992); US 2005/0276812 A1; and US 2005/016993 A1, the disclosures of which are expressly incorporated herein by reference. (2) Auristatin and Dolastatin

The drug part includes dolastatin, auristatin and their analogs and derivatives (US 5635483; US 5780588; US 5767237; US 6124431). Auristatin is a derivative of the marine mollusk compound Dolastatin-10. Although not intending to be bound by any particular theory, it has been shown that dolastatin and auristatin interfere with microtubule dynamics, GTP hydrolysis, nuclear and cell division (Woyke et al. (2001) Antimicrob. Agents and Chemother. 45() 12):3580-3584) and has anticancer (US 5663149) and antifungal activities (Pettit et al. (1998) Antimicrob. Agents Chemother. 42:2961-2965). The Dolastatin/Auristatin drug moiety can be linked to the antibody via the N (amino) end or C (carboxy) end of the peptide drug moiety (WO 02/088172; Doronina et al., (2003) Nature Biotechnology 21(7) :778-784; Francisco et al., (2003) Blood 102(4):1458-1465).

D_E

D_F 其中D_E 及D_F 之波形線指示與抗體或抗體-連接子組分共價連接之位點，且獨立地在各位置處： R² 係選自H及C₁ -C₈ 烷基； R³ 係選自H、C₁ -C₈ 烷基、C₃ -C₈ 碳環、芳基、C₁ -C₈ 烷基-芳基、C₁ -C₈ 烷基-(C₃ -C₈ 碳環)、C₃ -C₈ 雜環及C₁ -C₈ 烷基-(C₃ -C₈ 雜環)； R⁴ 係選自H、C₁ -C₈ 烷基、C₃ -C₈ 碳環、芳基、C₁ -C₈ 烷基-芳基、C₁ -C₈ 烷基-(C₃ -C₈ 碳環)、C₃ -C₈ 雜環及C₁ -C₈ 烷基-(C₃ -C₈ 雜環)； R⁵ 係選自H及甲基； 或R⁴ 及R⁵ 共同形成碳環且具有式-(CR^a R^b )_n -，其中R^a 及R^b 係獨立地選自H、C₁ -C₈ 烷基及C₃ -C₈ 碳環且n係選自2、3、4、5及6； R⁶ 係選自H及C₁ -C₈ 烷基； R⁷ 係選自H、C₁ -C₈ 烷基、C₃ -C₈ 碳環、芳基、C₁ -C₈ 烷基-芳基、C₁ -C₈ 烷基-(C₃ -C₈ 碳環)、C₃ -C₈ 雜環及C₁ -C₈ 烷基-(C₃ -C₈ 雜環)； 各R⁸ 係獨立地選自H、OH、C₁ -C₈ 烷基、C₃ -C₈ 碳環及O-(C₁ -C₈ 烷基)； R⁹ 係選自H及C₁ -C₈ 烷基； R¹⁰ 係選自芳基或C₃ -C₈ 雜環； Z為O、S、NH或NR¹² ，其中R¹² 為C₁ -C₈ 烷基； R¹¹ 係選自H、C₁ -C₂₀ 烷基、芳基、C₃ -C₈ 雜環、-(R¹³ O)_m -R¹⁴ 或-(R¹³ O)_m -CH(R¹⁵ )₂ ； m為1-1000範圍內之整數； R¹³ 為C₂ -C₈ 烷基； R¹⁴ 為H或C₁ -C₈ 烷基； R¹⁵ 在每次出現時皆獨立地爲H、COOH、-(CH₂ )_n -N(R¹⁶ )₂ 、-(CH₂ )_n -SO₃ H或-(CH₂ )_n -SO₃ -C₁ -C₈ 烷基； R¹⁶ 在每次出現時皆獨立地爲H、C₁ -C₈ 烷基或-(CH₂ )_n -COOH； R¹⁸ 係選自-C(R⁸ )₂ -C(R⁸ )₂ -芳基、-C(R⁸ )₂ -C(R⁸ )₂ -(C₃ -C₈ 雜環)及-C(R⁸ )₂ -C(R⁸ )₂ -(C₃ -C₈ 碳環)；且 n為0至6範圍內之整數。Exemplary auristatin embodiments include the disclosed in US 7498298 and US 7659241 N-linked monomethylauristatin single portion Rees statins D _E and D _F, to the disclosure of such patent is expressly incorporated by reference in its entirety herein by :

D _E

Wherein the waveform D _E D _F D _F and the line indicates the antibody or antibody - linker component is covalently connected to the site, and independently at each location: R ² is selected from H and C ₁ -C ₈ alkyl ; R ³ is selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ carbocyclic, aryl, C ₁ -C ₈ alkyl-aryl, C ₁ -C ₈ alkyl-(C _3- C ₈ carbocyclic ring), C ₃ -C ₈ heterocyclic ring and C ₁ -C ₈ alkyl-(C ₃ -C ₈ heterocyclic ring); R ⁴ is selected from H, C ₁ -C ₈ alkyl, C _3- C ₈ carbocyclic ring, aryl group, C ₁ -C ₈ alkyl-aryl group, C ₁ -C ₈ alkyl-(C ₃ -C ₈ carbocyclic ring), C ₃ -C ₈ heterocyclic ring and C ₁ -C ₈ Alkyl-(C ₃ -C ₈ heterocycle); R ⁵ is selected from H and methyl; or R ⁴ and R ⁵ together form a carbocyclic ring and have the formula -(CR ^a R ^b ) _n -, wherein R ^a and R ^b is independently selected from H, C ₁ -C ₈ alkyl and C ₃ -C ₈ carbocyclic ring and n is selected from 2, 3, 4, 5 and 6; R ⁶ is selected from H and C ₁ -C ₈ alkyl; R ⁷ is selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ carbocyclic, aryl, C ₁ -C ₈ alkyl-aryl, C ₁ -C ₈ alkyl-( C ₃ -C ₈ carbocyclic ring), C ₃ -C ₈ heterocycle and C ₁ -C ₈ alkyl-(C ₃ -C ₈ heterocycle); each R ⁸ is independently selected from H, OH, C _1- C ₈ alkyl, C ₃ -C ₈ carbocyclic and O-(C ₁ -C ₈ alkyl); R ⁹ is selected from H and C ₁ -C ₈ alkyl; R ¹⁰ is selected from aryl or C ₃ -C ₈ heterocycle; Z is O, S, NH or NR ¹² , wherein R ¹² is C ₁ -C ₈ alkyl; R ¹¹ is selected from H, C ₁ -C ₂₀ alkyl, aryl, C _3- C ₈ heterocycle, -(R ¹³ O) _m -R ¹⁴ or -(R ¹³ O) _m -CH(R ¹⁵ ) ₂ ; m is an integer in the range of 1-1000; R ¹³ is C ₂ -C ₈ alkane R ¹⁴ is H or C ₁ -C ₈ alkyl; R ¹⁵ is independently H, COOH, -(CH ₂ ) _n -N(R ¹⁶ ) ₂ , -(CH ₂ ) _n each time it appears -SO ₃ H or -(CH ₂ ) _n -SO ₃ -C ₁ -C ₈ alkyl; each occurrence of R ¹⁶ is independently H, C ₁ -C ₈ alkyl or -(CH ₂ ) _n -COOH; R ¹⁸ is selected from -C(R ⁸ ) ₂ -C(R ⁸ ) ₂ -aryl, -C(R ⁸ ) ₂ -C(R ⁸ ) ₂ -(C ₃ -C ₈ heterocycle) And -C(R ⁸ ) ₂ -C(R ⁸ ) ₂ -(C ₃ -C ₈ carbocyclic ring); and n is an integer in the range of 0 to 6.

In one embodiment, R ³ , R ⁴ and R ^{7 are} independently isopropyl or sec-butyl and R ⁵ is -H or methyl. In an exemplary embodiment, R ³ and R ⁴ are each isopropyl, R ⁵ is -H, and R ⁷ is sec-butyl.

In another embodiment, R ² and R ⁶ are each methyl, and R ⁹ is -H.

In another embodiment, each occurrence of R ⁸ is -OCH ₃ .

In an exemplary embodiment, R ³ and R ⁴ are each isopropyl, R ² and R ⁶ are each methyl, R ⁵ is -H, R ⁷ is sec-butyl, and R ⁸ is each occurrence All are -OCH ₃ , and R ⁹ is -H.

In one embodiment, Z is -O- or -NH-.

In one embodiment, R ¹⁰ is an aryl group.

In an exemplary embodiment, R ¹⁰ is -phenyl.

In an exemplary embodiment, when Z is -O-, R ¹¹ is -H, methyl, or tertiary butyl.

In one embodiment, when Z is -NH, R ¹¹ is -CH(R ¹⁵ ) ₂ , wherein R ¹⁵ is -(CH ₂ ) _n -N(R ¹⁶ ) ₂ , and R ¹⁶ is -C _1- C ₈ alkyl or -(CH ₂ ) _n -COOH.

In another embodiment, when Z is -NH, R ¹¹ is -CH(R ¹⁵ ) ₂ , wherein R ¹⁵ is -(CH ₂ ) _n -SO ₃ H.

MMAEExemplary auristatin with one embodiment of formula D _E is MMAE, wherein the wavy line indicates the anti-CD79b immunoconjugate composition of the linker (L) of covalently linked:

MMAE

MMAFAn exemplary auristatin example of formula _DF is MMAF, where the wavy line indicates the covalent connection to the linker (L) of the anti-CD79b immunoconjugate:

MMAF

Other exemplary embodiments include a monomethylvaline compound (WO 2007/008848) with a carboxyl group modification at the C-terminus of the pentapeptide auristatin drug moiety and at the C-terminus of the pentapeptide auristatin drug moiety Monomethylvaline compound with modified side chain of phenylalanine (WO 2007/008603).

Ab-MC-vc-PAB-MMAF

Ab-MC-vc-PAB-MMAE

Ab-MC-MMAE

Ab-MC-MMAFNon-limiting exemplary embodiments of anti-CD79b immunoconjugates of formula I comprising MMAE or MMAF and various linker components have the following structures and abbreviations (wherein "Ab" is an anti-CD79b antibody; p is from 1 to about 8, "Val -Cit" is valine-citrulline dipeptide; and "S" is sulfur atom):

Ab-MC-vc-PAB-MMAF

Ab-MC-vc-PAB-MMAE

Ab-MC-MMAE

Ab-MC-MMAF

In certain embodiments, the anti-CD79b immunoconjugate comprises the structure of Ab-MC-vc-PAB-MMAE, wherein p is, for example, about 1 to about 8; about 2 to about 7; about 3 to about 5; about 3 to About 4; or about 3.5. In some embodiments, the anti-CD79b immunoconjugate is huMA79bv28-MC-vc-PAB-MMAE, such as an anti-CD79b immunoconjugate comprising the structure of MC-vc-PAB-MMAE, wherein p is, for example, about 1 to about 8; about 2 to about 7; about 3 to about 5; about 3 to about 4; or about 3.5, wherein the anti-CD79 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 36, and wherein the light chain comprises SEQ ID NO: 35 amino acid sequence. In some embodiments, the anti-CD79b immunoconjugate is polatuzumab vedotin (CAS number 1313206-42-6).

Non-limiting exemplary embodiments of the anti-CD79b immunoconjugates of Formula I comprising MMAF and various linker components further include Ab-MC-PAB-MMAF and Ab-PAB-MMAF. Immunoconjugates comprising MMAF linked to an antibody by a non-proteolytically cleavable linker have been shown to have similar activity to immunoconjugates comprising MMAF linked to an antibody by a proteolytically cleavable linker (Doronina et al., (2006) Bioconjugate Chem. 17:114-124). In some of these embodiments, it is believed that drug release is achieved by the degradation of the antibody in the cell.

Generally, peptide-based drug moieties can be prepared by forming peptide bonds between two or more amino acids and/or peptide fragments. For example, these peptide bonds can be prepared according to a liquid phase synthesis method ( see, for example, E. Schröder and K. Lübke, "The Peptides", Volume 1, Pages 76-136, 1965, Academic Press). In some embodiments, the auristatin/dolastatin drug portion can be prepared according to the following methods: US 7498298; US 5635483; US 5780588; Pettit et al. (1989) J. Am. Chem. Soc . 111:5463-5465 ; Pettit et al. (1998) Anti-Cancer Drug Design 13:243-277; Pettit, GR et al. Synthesis , 1996, 719-725; Pettit et al. (1996) J. Chem. Soc. Perkin Trans. 1 5:859 -863; and Doronina (2003) Nat. Biotechnol. 21(7):778-784.

In some embodiments, formula D _E such as MMAE and _DF such as the auristatin/dolastatin drug portion of MMAF, and drug-linker intermediates and derivatives thereof, such as MC-MMAF, MC-MMAE, MC -vc-PAB-MMAF and MC-vc-PAB-MMAE can use US 7498298; Doronina et al. (2006) Bioconjugate Chem. 17:114-124; and Doronina et al. (2003) Nat. Biotech. 21:778-784 Prepared by the method described in and then combined with the antibody of interest. (3) calicheamicin

In some embodiments, the anti-CD79b immunoconjugate comprises an anti-CD79b antibody bound to one or more calicheamicin molecules. Antibiotics of the calicheamicin family and their analogs can cause double-stranded DNA breaks at concentrations lower than picomoles (Hinman et al., (1993) Cancer Research 53:3336-3342; Lode et al., (1998) Cancer Research 58:2925-2928). Calicheamicin has intracellular sites of action, but in some cases it is not easy to cross the plasma membrane. Therefore, in some embodiments, cellular uptake of these agents through antibody-mediated internalization can greatly enhance their cytotoxic effects. Non-limiting exemplary methods for preparing anti-CD79b antibody immunoconjugates with a calicheamicin drug moiety are described in, for example, US 5712374; US 5714586; US 5739116; and US 5767285. (4) Other drugs

In some embodiments, the anti-CD79b immunoconjugate comprises geldanamycin (Mandler et al. (2000) J. Nat. Cancer Inst. 92(19): 1573-1581; Mandler et al. (2000) Bioorganic & Med. Chem. Letters 10:1025-1028; Mandler et al. (2002) Bioconjugate Chem. 13:786-791); and/or enzyme-active toxins and fragments thereof, including but not limited to diphtheria A chain, non-binding active fragments of diphtheria toxin , Exotoxin A chain (from Pseudomonas aeruginosa), Ricin A chain, Acacia toxin A chain, Capsule root toxin A chain, α-Chodophyllum, Aleurites fordii protein, Dianthus Dianthin protein, Phytolaca americana protein (PAPI, PAPII and PAP-S), momordica charantia inhibitor, saponin, crotonin, sapaonaria officinalis inhibitor, white tree Toxin (gelonin), mitogellin (mitogellin), restrictocin (restrictocin), phenomycin (phenomycin), inomycin (enomycin) and trichothecenes (tricothecene). See, for example, WO 93/21232.

The drug portion also includes compounds with nuclear degrading activity ( for example, ribonuclease or DNA endonuclease).

In certain embodiments, the anti-CD79b immunoconjugate contains highly radioactive atoms. A variety of radioisotopes can be used to produce radioactively conjugated antibodies. Examples include At ²¹¹ , I ¹³¹ , I ¹²⁵ , Y ⁹⁰ , Re ¹⁸⁶ , Re ¹⁸⁸ , Sm ¹⁵³ , Bi ²¹² , P ³² , Pb ²¹² and radioactive isotopes of Lu. In some embodiments, when the anti-CD79b immunoconjugate is used for detection, it may contain radioactive atoms used in scintigraphic research, such as Tc ⁹⁹ or I ¹²³ ; or used in nuclear magnetic resonance (NMR) imaging (also known as Magnetic resonance imaging (MRI) spin labels, such as zirconium-89, iodine-123, iodine-131, indium-111, fluorine-19, carbon-13, nitrogen-15, oxygen-17, gamma, manganese or iron. Zirconium-89 can be complexed with various metal chelating agents and bound to antibodies, for example for PET imaging (WO 2011/056983).

Radioactive labels or other labels can be incorporated into the anti-CD79b immunoconjugate in a known manner. For example, peptides can be biosynthesized or chemically synthesized using suitable amino acid precursors containing, for example, one or more fluorine-19 atoms in place of one or more hydrogens. In some embodiments, labels such as Tc ⁹⁹ , I ¹²³ , Re ¹⁸⁶ , Re ¹⁸⁸ and In ¹¹¹ can be linked via cysteine residues in the anti-CD79b antibody. In some embodiments, yttrium-90 can be linked via lysine residues of the anti-CD79b antibody. In some embodiments, the IODOGEN method (Fraker et al., (1978) Biochem. Biophys. Res. Commun. 80: 49-57) can be used to incorporate iodine-123. "Monoclonal Antibodies in Immunoscintigraphy" (Chatal, CRC Press 1989) describes certain other methods.

In certain embodiments, the anti-CD79b immunoconjugate may comprise an anti-CD79b antibody that binds to a prodrug activating enzyme. In some of these embodiments, prodrug activating enzymes convert prodrugs ( e.g., peptidyl chemotherapeutics, see WO 81/01145) into active drugs, such as anticancer drugs. In some embodiments, these immunoconjugates are suitable for use in antibody-dependent enzyme-mediated prior drug therapy ("ADEPT"). Enzymes that can bind to anti-CD79b antibodies include (but are not limited to) alkaline phosphatase, which is suitable for converting phosphate-containing prodrugs into free drugs; arylsulfatase, which is suitable for converting sulfuric acid-containing prodrugs into free drugs ; Cytosine deaminase, which is suitable for converting non-toxic 5-fluorocytosine into anti-cancer drugs, 5-fluorouracil; proteases, such as serratia protease, thermolysin, subtilisin (subtilisin), carboxypeptidase, and cathepsin (such as cathepsin B and L), which are suitable for converting peptide-containing prodrugs into free drugs; D-alanine carboxypeptidase, which is suitable for conversion containing D-amine groups Acid substituent prodrug; carbohydrate lyase, such as β-galactosidase (galactosidase) and neuraminidase, which are suitable for converting glycosylated prodrugs into free drugs; β-endosidase ( lactamase), which is suitable for converting drugs derived from β-lactam into free drugs; and penicillin amidase, such as penicillin V amidase and penicillin G amidase, which is suitable for separating at its amine nitrogen Drugs derived with phenoxyacetoxy or phenylacetoxy are converted into free drugs. In some embodiments, the enzyme can be covalently bound to the antibody by recombinant DNA technology well known in the art. See, for example, Neuberger et al., Nature 312:604-608 (1984). D. Drug loading

The drug loading is represented by p, which is the average number of drug portions corresponding to each anti-CD79b antibody in the formula I molecule. The drug loading can be in the range of 1 to 20 drug parts (D) per antibody. The anti-CD79b immunoconjugates of formula I include a collection of anti-CD79b antibodies bound to a certain range (1 to 20) of drug moieties. The average number of drug parts corresponding to each anti-CD79b antibody in the anti-CD79b immunoconjugate preparation obtained by the binding reaction can be characterized by conventional means such as mass spectrometry, ELISA analysis and HPLC. The quantitative distribution of anti-CD79b immunoconjugates denoted by p can also be determined. In some cases, separation, purification, and characterization of homogeneous anti-CD79b immunoconjugates with p to a certain value from anti-CD79b immunoconjugates with other drug loadings can be achieved by means such as reverse phase HPLC or electrophoresis.

For some anti-CD79b immunoconjugates, p can be limited by the number of attachment sites on the anti-CD79b antibody. For example, when the connection is a cysteine thiol as in certain exemplary embodiments above, the anti-CD79b antibody may only have one or several cysteine thiol groups, or may only have One or several thiol groups of the linker with sufficient reactivity. In some embodiments, higher drug loading (e.g., p>5) can lead to aggregation, insolubility, toxicity, or reduced cell permeability of certain anti-CD79b immunoconjugates. In certain embodiments, the average drug loading of the anti-CD79b immunoconjugate ranges from 1 to about 8; from about 2 to about 6; from about 3 to about 5; or from about 3 to about 4. In fact, it has been shown that for certain antibody-drug conjugates, the optimal ratio of the drug portion of each antibody can be less than 8, and can range from about 2 to about 5 (US 7498298). In certain embodiments, the optimal ratio of the drug portion of each antibody is about 3 to about 4. In certain embodiments, the optimal ratio of the drug portion of each antibody is about 3.5.

In certain embodiments, the portion of the drug that is less than the theoretical maximum is bound to the anti-CD79b antibody during the binding reaction. Antibodies may contain, for example, lysine residues that do not react with drug-linker intermediates or linker reagents, as discussed below. Generally speaking, antibodies do not contain many free and reactive cysteine thiol groups that can be linked to the drug moiety; in fact, most of the cysteine thiol residues in antibodies exist as disulfide bridges. In certain embodiments, reducing agents such as dithiothreitol (DTT) or tricarbonyl ethyl phosphine (TCEP) can be used to reduce the anti-CD79b antibody under partial or complete reduction conditions to produce reactive cysteine Thiol group. In certain embodiments, the anti-CD79b antibody is subjected to denaturing conditions to reveal reactive nucleophilic groups, such as lysine or cysteine.

The loading of anti-CD79b immunoconjugates (drug/antibody ratio) can be controlled in different ways and, for example, by: (i) limiting the molar excess of the drug-linker intermediate or linker reagent relative to the antibody, (ii) Limit the binding reaction time or temperature, and (iii) part or limit the reducing conditions for cysteine thiol modification.

It should be understood that when more than one nucleophilic group reacts with a drug-linker intermediate or a linker reagent, the resulting product is an anti-CD79b immunoconjugate compound with a certain distribution of one or more drug moieties linked to the anti-CD79b antibody mixture. The average number of drugs corresponding to each antibody can be calculated from the mixture by double ELISA antibody analysis that is specific for the antibody and specific for the drug. Individual anti-CD79b immunoconjugate molecules can be identified in the mixture by mass spectrometry and separated by HPLC, such as hydrophobic interaction chromatography ( see, for example, McDonagh et al. (2006) Prot. Engr. Design & Selection 19(7) :299-307; Hamblett et al. (2004) Clin. Cancer Res. 10:7063-7070; Hamblett, KJ et al. "Effect of drug loading on the pharmacology, pharmacokinetics, and toxicity of an anti-CD30 antibody-drug conjugate, "Abstract No. 624, American Association for Cancer Research, 2004 Annual Meeting, March 27-31, 2004, Proceedings of the AACR, Volume 45, March 2004; Alley, SC et al. "Controlling the location of drug attachment in antibody-drug conjugates," Abstract No. 627, American Association for Cancer Research, 2004 Annual Meeting, March 27-31, 2004, Proceedings of the AACR, Volume 45, March 2004). In certain embodiments, homogeneous anti-CD79b immunoconjugates with a single loading value can be separated from the binding mixture by electrophoresis or chromatography. E. Method for preparing anti- CD79b immunoconjugate

The anti-CD79b immunoconjugate of formula I can be prepared by a number of methods using organic chemical reactions, conditions and reagents known to those skilled in the art, including but not limited to, for example: (1) nucleophilic group and bivalent anti-CD79b antibody The linker reagent reacts to form Ab-L via a covalent bond, and then reacts with the drug moiety D; and (2) the nucleophilic group of the drug moiety reacts with the divalent linker reagent to form DL via a covalent bond, and then with the anti- The nucleophilic group of the CD79b antibody reacts. An exemplary method for preparing an anti-CD79b immunoconjugate of formula I via the following route is described in US 7498298, which is expressly incorporated herein by reference.

The nucleophilic groups on the antibody include (but are not limited to): (i) N-terminal amine group, (ii) side chain amine group, such as lysine, (iii) side chain thiol group, such as cysteine, And (iv) a sugar hydroxyl or amine group, wherein the antibody is glycosylated. The amine group, thiol group and hydroxyl group have nucleophilicity and can react with the linker part and the electrophilic group on the linker reagent including the following to form a covalent bond: (i) active ester, such as NHS ester, HOBt ester, Halogenated formate and acid halides; (ii) alkyl and benzyl halides, such as haloacetamide; and (iii) aldehyde, ketone, carboxyl and maleimide groups. Certain antibodies have reducible interchain disulfides, which are cysteine bridges. The anti-CD79b antibody can be made reactive with the linker reagent by treatment with a reducing agent such as DTT (dithiothreitol) or tricarbonyl ethyl phosphine (TCEP) to completely or partially reduce the anti-CD79b antibody. Each cysteine bridge will therefore theoretically form two reactive thiol nucleophiles. Other nucleophilic groups can be modified by lysine residues, for example by reacting lysine residues with 2-iminothiocyclopentane (Traut's reagent) to make the amine Converted into thiol, and introduced into the anti-CD79b antibody. Reactive thiol groups can also be introduced into anti-CD79b antibodies by introducing one, two, three, four, or more than four cysteine residues ( e.g., by preparing one or more unnatural cysteine Variant antibodies with amino acid residues).

The anti-CD79b immunoconjugates described herein can also be produced by the reaction between an electrophilic group (such as an aldehyde or ketone carbonyl) on the anti-CD79b antibody and a nucleophilic group on a linker reagent or drug. Suitable nucleophilic groups on the linker reagent include, but are not limited to, hydrazine, oxime, amine, hydrazine, thiosemicarbhydrazone, hydrazine carboxylate and arylhydrazine. In one embodiment, the anti-CD79b antibody is modified to introduce an electrophilic moiety capable of reacting with a nucleophilic substituent on a linker reagent or drug. In another embodiment, the sugar of the glycosylated anti-CD79b antibody can be oxidized with a periodate oxidizing reagent, for example, to form an aldehyde or ketone group that can react with the linker reagent or the amine group of the drug moiety. The resulting imine Schiff base group can form a stable linkage, or can be reduced, for example, by a borohydride reagent to form a stable amine linkage. In one embodiment, the reaction of the carbohydrate portion of the glycosylated anti-CD79b antibody with galactose oxidase or sodium metaperiodate can produce a carbonyl group (aldehyde and aldehyde) in the anti-CD79b antibody that can react with appropriate groups on the drug. Ketone) (Hermanson, Bioconjugate Techniques). In another embodiment, an anti-CD79b antibody containing an N-terminal serine or threonine residue can be reacted with sodium metaperiodate to produce an aldehyde instead of the first amino acid (Geoghegan and Stroh, (1992) Bioconjugate Chem. 3:138-146; US 5362852). The aldehyde can react with the drug moiety or the linker nucleophile.

Exemplary nucleophilic groups on the drug moiety include but are not limited to: amine, thiol, hydroxyl, hydrazine, oxime, hydrazine, thiosemicarbhydrazone, hydrazine carboxylate and aryl hydrazine groups, which can be combined with The following linker moieties and the electrophilic group on the linker reagent react to form a covalent bond: (i) active esters, such as NHS esters, HOBt esters, haloformates and acid halides; (ii) alkyl groups and Benzyl halides, such as haloacetamide; (iii) aldehyde, ketone, carboxyl, and maleimide groups.

Non-limiting exemplary crosslinker reagents that can be used to prepare anti-CD79b immunoconjugates are described herein in the section entitled "Exemplary Linkers." The method of using these crosslinker reagents to connect two parts (including the protein part and the chemical part) is known in the art. In some embodiments, a fusion protein containing an anti-CD79b antibody and a cytotoxic agent can be prepared, for example, by recombinant technology or peptide synthesis. Recombinant DNA molecules may include regions encoding antibodies and cytotoxic portions of the conjugate, which regions are adjacent to each other or separated by regions encoding linker peptides that do not destroy the desired properties of the conjugate. In another example, an anti-CD79b antibody can bind to a "receptor" (such as streptavidin) for use in tumor pre-targeting, where the antibody-receptor conjugate is administered to the patient, followed by A scavenger is used to remove unbound conjugates from the circulation and then a "ligand" (such as avidin) that binds to a cytotoxic agent (such as a drug or radionucleotide) is administered. Other details about the anti-CD79b immunoconjugates are provided in US Patent No. 8545850 and WO/2016/049214, the contents of which are expressly incorporated herein by reference in their entirety. V. Alkylating agent

Alkylating agents are a class of antitumor drugs or anticancer drugs that act by inhibiting the transcription of DNA into RNA and thereby stopping protein synthesis. Alkylation reagents replace hydrogen atoms on DNA with alkyl groups (C _n H _2n+1 ), which leads to the formation of cross-links in DNA strands, which leads to DNA strand breaks, resulting in abnormal base pairing, inhibiting cell division, and ultimately cell death. This effect occurs in all cells, but rapidly dividing cells, such as cancer cells, are usually the most sensitive to the effects of alkylating agents

Alkylating agents are generally divided into six categories: (1) Nitrogen mustards, which include, but are not limited to, for example, methyl ethylamine, cyclophosphamide, ifosphamide, bendamustine, melphalan, and phenylbutyric acid Nitrogen mustard; (2) vinylamine and methyleneamine derivatives, including but not limited to, for example, hexamethylmelamine and Thiotepa; (3) alkyl sulfonates, including but not limited to, for example, busulfan; (4) Nitrosoureas, including but not limited to carmustine and lomustine; (5) triazene, including but not limited to dacarbazine and procarbazine, temozolomide; (6) platinum-containing anti Tumor agents include, but are not limited to, for example, cisplatin, carboplatin, and oxaliplatin. Any known alkylating agents (including but not limited to those listed above) can be used in the treatment methods provided herein.

Bendamustine is an exemplary alkylating agent used in the methods described herein. The chemical name of bendamustine is 4-(5-(bis(2-chloroethyl)amino)-1-methyl-1H-benzo[d]imidazol-2-yl)butyric acid, benda Mustine has the following structural formula:

Bendamustine (CAS registration #16506-27-7) has a molecular formula of C ₁₆ H ₂₁ Cl ₂ N ₃ O ₂ and a molecular weight of 358.263 g/mol. Bendamustine is a bifunctional dichloromethyldiethylamine derivative containing a purine-like benzimidazole ring. Bendamustine can be used as a powder for solution and solution dosage forms.

HClIn some embodiments, the alkylating agent used in the methods described herein is a salt or solvate of bendamustine. In some embodiments, the bendamustine salt is bendamustine-HCl (CAS#3543-75-7), its molecular formula is C ₁₆ H ₂₁ Cl ₂ N ₃ O ₂ ·HCl, and the molecular weight is 394.72 g /mol.

HCl

Bendamustine-HCl is commercially available under the trade names BENDEKA, TREANDA, TREAKISYM, RIBOMUSTIN, LEVACT, MUSTIN, etc. VI. Anti- CD20 agents

According to the binding characteristics and biological activity of anti-CD20 antibody and CD20 antigen, two types of anti-CD20 antibodies (type I and type II anti-CD20 antibodies) can be distinguished, according to Cragg, MS et al., Blood 103 (2004) 2738-2743; and Cragg, MS, et al., Blood 101 (2003) 1045-1052, see table C. Table C : Properties of Type I and Type II Anti- CD20 Antibodies

Examples of type I anti-CD20 antibodies include, for example, rituximab, HI47 IgG3 (ECACC, fusion tumor), 2C6 IgG1 (as disclosed in WO 2005/103081), 2F2 IgG1 (such as WO 2004/035607 and WO 2005/103081) Disclosed) and 2H7 IgG1 (as disclosed in WO 2004/056312).

In some embodiments, the anti-CD20 antibody system rituximab of the treatment methods provided herein is used. In some embodiments, rituximab (reference antibody; an example of type I anti-CD20 antibody) is a genetically engineered chimeric monoclonal antibody containing human γ1 murine constant domain, which is directed against the human CD20 antigen. However, the antibody is not glycosylated, and is not non-trehalose, so it has a trehalose content of at least 85%. The chimeric antibody contains the human γ1 constant domain and was identified by the name "C2B8" in US 5,736,137 (Andersen et al. ) issued to IDEC Pharmaceuticals Corporation on April 17, 1998. Rituximab is approved for the treatment of patients with relapsed or refractory low-grade or follicular CD20-positive, B-cell non-Hodgkin lymphoma. In vitro studies of the mechanism of action have shown that rituximab has human complement-dependent cytotoxicity (CDC) (Reff, ME et al., Blood 83(2) (1994) 435-445). In addition, it has shown activity in the assay to measure antibody-dependent cellular cytotoxicity (ADCC).

In some embodiments, the anti-CD20 antibody system used in the treatment methods provided herein is a non-trehalose anti-CD20 antibody.

Examples of type II anti-CD20 antibodies include, for example, the humanized B-Ly1 antibody IgG1 (chimeric humanized IgG1 antibody disclosed in WO 2005/044859), 11B8 IgG1 (as disclosed in WO 2004/035607) and AT80 IgG1. Generally, type II anti-CD20 antibodies of the IgG1 isotype show characteristic CDC properties. Compared with type I antibodies of IgG1 isotype, type II anti-CD20 antibodies have a reduced CDC (if IgG1 isotype). In some embodiments, type II anti-CD20 antibodies, such as GA101 antibodies, have increased antibody-dependent cellular cytotoxicity (ADCC). In some embodiments, the type II anti-CD20 antibody is more preferably a non-trehalosylated humanized B-Ly1 antibody as described in WO 2005/044859 and WO 2007/031875.

In some embodiments, the anti-CD20 anti-system GA101 antibody used in the treatment methods provided herein. In some embodiments, the GA101 antibody system as used herein refers to any of the following antibodies that bind to human CD20: (1) Antibodies comprising each of the following: HVR- comprising the amino acid sequence of SEQ ID NO: 5 H1, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 6, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 7, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 8, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 9 and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 10; (2) Antibody comprising the amino acid of SEQ ID NO: 11 The VH domain of the sequence and the VL domain containing the amino acid sequence of SEQ ID NO: 12, (3) an antibody containing the amino acid sequence of SEQ ID NO: 13 and the amino acid sequence of SEQ ID NO: 14; (4) An antibody called atolizumab, or (5) an amine containing at least 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO: 13 The antibody contains an amino acid sequence that has at least 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO: 14. In one example, the GA101 anti-system IgG1 isotype antibody.

In some embodiments, the anti-CD20 antibody system humanized B-Ly1 antibody used in the treatment methods provided herein. In some embodiments, the humanized B-Ly1 antibody system refers to the humanized B-Ly1 antibody disclosed in WO 2005/044859 and WO 2007/031875, which is chimeric with a human constant domain derived from IgG1 and developed in humans. After sourceization, it was obtained from mouse monoclonal anti-CD20 antibody B-Ly1 (variable region of mouse heavy chain (VH): SEQ ID NO: 3; variable region of mouse light chain (VL): SEQ ID NO: 4- See Poppema, S. and Visser, L., Biotest Bulletin 3 (1987) 131-139) (see WO 2005/044859 and WO 2007/031875). The humanized B-Ly1 antibody is disclosed in detail in WO 2005/044859 and WO 2007/031875.

In some embodiments, the humanized B-Ly1 antibody has a heavy chain variable region (VH) selected from the group of SEQ ID NOs: 15-16 and 40-55 (corresponding to WO 2005/044859 and WO 2007/031875 B-HH2 to B-HH9 and B-HL8 to B-HL17). In some embodiments, the variable domain is selected from the group consisting of SEQ ID NO: 15, 16, 42, 44, 46, 48, and 50 (corresponding to B-HH2, BHH in WO 2005/044859 and WO 2007/031875 -3, B-HH6, B-HH8, B-HL8, B-HL11 and B-HL13). In some embodiments, the humanized B-Ly1 antibody has a light chain variable region (VL) of SEQ ID NO: 55 (corresponding to B-KV1 of WO 2005/044859 and WO 2007/031875). In some embodiments, the humanized B-Ly1 antibody has a heavy chain variable region (VH) of SEQ ID NO: 42 (corresponding to B-HH6 of WO 2005/044859 and WO 2007/031875) and SEQ ID NO: The light chain variable region (VL) of 55 (corresponding to B-KV1 of WO 2005/044859 and WO 2007/031875). In some embodiments, humanized B-Ly1 anti-system IgG1 antibodies. According to the procedures described in WO 2005/044859, WO 2004/065540, WO 2007/031875, Umana, P. et al. Nature Biotechnol. 17 (1999) 176-180 and WO 99/154342, these non-trehalose-based The humanized B-Ly1 antibody is glycosylated (GE) in the Fc region. In some embodiments, the non-trehalosylated humanized B-Ly1 that is glycosylated is B-HH6-B-KV1 GE. In some embodiments, the anti-CD20 antibody system atolizumab (Recommended INN, WHO Drug Information, Volume 26, Issue 4, 2012, Page 453). As used herein, atolizumab is synonymous with GA101 or RO5072759. This replaces all previous versions (for example, Volume 25, Issue 1, 2011, Pages 75-76), formerly known as Afutuzhu (Recommended INN, WHO Drug Information, Volume 23, Issue 2 , 2009, page 176; Volume 22, Issue 2, 2008, page 124). In some embodiments, the humanized B-Ly1 antibody system comprises an antibody containing the heavy chain of the amino acid sequence of SEQ ID NO: 17 and the light chain of the amino acid sequence of SEQ ID NO: 18, or this antibody The antigen-binding fragment. In some embodiments, the humanized B-Ly1 antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising the three heavy chain CDRs of SEQ ID NO: 17, the light chain variable The region contains the three light chain CDRs of SEQ ID NO:18.

In some embodiments, the humanized B-Ly1 antibody system is non-trehalosylated and glycosylated humanized B-Ly1. These glycosylated humanized B-Ly1 antibodies have altered glycosylation patterns in the Fc region, preferably with reduced levels of trehalose residues. In some embodiments, the amount of trehalose is about 60% or less of the total amount of oligosaccharides at Asn297 (in one embodiment, the amount of trehalose is about 40% to about 60%, in another embodiment , The amount of trehalose is about 50% or less, in another embodiment, the amount of trehalose is about 30% or less). In some embodiments, the oligosaccharides in the Fc region are divided equally. These glycosylated humanized B-Ly1 antibodies have increased ADCC.

MFI係平均螢光強度。如本文所用之「Cy5標記比率」係指每個分子抗體之Cy5標記分子之數目。As described in Example 2, in a FACSArray (Becton Dickinson) with Raji cells (ATCC-No. CCL-86), the anti-CD20 antibody that binds to Cy5 and rituximab that bind to Cy5 are used, by Direct immunofluorescence measurement (mean fluorescence intensity (MFI) is measured to determine the ratio of anti-CD20 antibody binding ability to CD20 on Raji cells (ATCC-No. CCL-86) compared with rituximab) ", and calculated as follows: Ratio of CD20 binding capacity to Raji cell (ATCC-No. CCL-86) =

MFI is the average fluorescence intensity. The "Cy5 labeling ratio" as used herein refers to the number of Cy5 labeling molecules per antibody molecule.

Typically, the type II anti-CD20 antibody has 0.3 to 0.6, and in one embodiment, 0.35 to 0.55, in another embodiment, 0.4 to 0.5 of the second anti-CD20 antibody compared to rituximab Ratio to CD20 binding capacity on Raji cells (ATCC-No. CCL-86).

"An antibody with increased antibody-dependent cellular cytotoxicity (ADCC)" means that the antibody (the term is defined herein) has an increased ADCC determined by any suitable method known to those of ordinary skill in the art.

The following describes an exemplary acceptable in vitro ADCC assay: 1) The assay uses target cells known to express the target antigen recognized by the antigen binding region of the antibody; 2) The assay uses blood from a randomly selected healthy donor Peripheral blood mononuclear cells (PBMC) isolated from PBMC were used as effector cells; 3) The assay was performed according to the following protocol: i) PBMC was separated using standard density centrifugation procedures and suspended in RPMI cell culture medium at 5×10 ⁶ cells/ml Medium; ii) Culturing target cells by standard tissue culture methods, harvesting from the exponential growth phase with viability higher than 90%, washing in RPMI cell culture medium, labeling with 100 microcuries of ⁵¹ Cr, and washing twice with cell culture medium times at 10 ⁵ cells / ml density of the resuspended cell culture medium; iii) 100 microliters of the final target cell suspension above was transferred to each well of the microtiter plate wells 96; IV) to the antibody in a cell Dilute serially from 4000 ng/ml to 0.04 ng/ml in the culture medium, and add 50 microliters of the obtained antibody solution to the target cells in a 96-well microtiter plate to test various antibody concentrations covering the entire concentration range in triplicate; v) For the maximum release (MR) control, the other 3 wells in the plate containing the labeled target cells received 50 microliters of 2% (VN) non-ionic detergent in water (Nonidet, Sigma, St. Louis) instead of Antibody solution (point iv above); vi) For spontaneous release (SR) control, the other 3 wells in the plate containing the labeled target cells receive 50 μl of RPMI cell culture medium instead of antibody solution (point iv above) Vii) Then centrifuge the 96-well microtiter plate at 50×g for 1 minute and incubate at 4°C for 1 hour; viii) add 50 microliters of PBMC suspension (the i-th point above) to each well to produce 25:1 effector: target cell ratio, and place the plate in an incubator at 37°C under a 5% CO2 atmosphere for 4 hours; ix) Harvest the cell-free supernatant from each well and use γ Counter quantitative experiment released radioactivity (ER); x) Calculate the specific lysis percentage of each antibody concentration according to the formula (ER-MR)/(MR-SR) x 100, where ER is the average radioactivity quantified for the antibody concentration (See point ix above), MR is the average radioactivity quantified against the MR control (see point V above) (see point ix above), and SR is the average radioactivity quantified against the SR control (see point vi above) (See point ix above); 4) "Increase ADCC" is defined as the increase in the maximum percentage of specific lysis observed within the antibody concentration range of the above test, and/or achieve the above test The reduction in antibody concentration required for half of the maximum percentage of specific lysis observed within the antibody concentration range. In one embodiment, the increase in ADCC is mediated by the above assay method using the same standard production, purification, preparation and storage methods known to those skilled in the art, and the same antibody produced by the same type of host cell. ADCC measured, except that the comparative antibody (lack of increased ADCC) has not been engineered to over-express GnTIII and/or engineered to have reduced performance of the trehalosyltransferase 8 (FUT8) gene ( e.g. , including, Produced by engineered host cells with the FUT8 group removed).

In some embodiments, "increased ADCC" can be obtained by, for example, mutation and/or glycosylation of the antibodies. In some embodiments, the anti-CD20 antibody is glycosylated to have biantennary oligosaccharides linked to the Fc region of the antibody, which oligosaccharides are equally divided by GlcNAc. In some embodiments, by using a host cell that lacks protein trehalosylation ( e.g. , Lec13 CHO cell or α-1,6-trehalosyltransferase gene (FUT8) deletion or FUT gene expression knockdown cell ), the anti-CD20 antibody is glycosylated to lack trehalose on the carbohydrate linked to the Fc region. In some embodiments, the anti-CD20 antibody sequence has been engineered in its Fc region to enhance ADCC. In some embodiments, such engineered anti-CD20 antibody variants comprise an Fc region with one or more amino acid substitutions (EU numbering of residues) at positions 298, 333, and/or 334 of the Fc region.

In some embodiments, the term "complement dependent cytotoxicity (CDC)" refers to the lysis of human cancer target cells by an antibody according to the present invention in the presence of complement. CDC can be measured by treating a cell preparation expressing CD20 with the anti-CD20 antibody of the present invention in the presence of complement. If the antibody induces lysis (cell death) of 20% or more tumor cells after 4 hours at a concentration of 100 nM, CDC is found. In some embodiments, tumor cells labeled with ⁵¹ Cr or Eu are used for the assay and the released ⁵¹ Cr or Eu is measured. Controls include growing tumor target cells with complement but not with antibodies.

In some embodiments, the anti-CD20 antibody system monoclonal antibody, such as a human antibody. In one embodiment, the anti-CD20 antibody is an antibody fragment, such as Fv, Fab, Fab', scFv, mini diabody, or F(ab') ₂ fragment. In another embodiment, the anti-CD20 antibody is a substantially full-length antibody, such as an IgG1 antibody, an IgG2a antibody, or other antibody class or isotype as defined herein. VII. Antibodies

In some embodiments, the antibodies used in the treatment methods provided herein (for example, anti-CD79b antibodies or anti-CD20 antibodies) may incorporate any of the features described below, alone or in combination. A. Antibody affinity

In certain embodiments, the antibody used in the treatment methods provided herein (eg, anti-CD79b antibody or anti-CD20 antibody) has ≤1 μM, ≤100 nM, ≤50 nM, ≤10 nM, ≤5 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM, or ≤0.001 nM, and optionally ≥10 ^-13 M dissociation constant (Kd). (For example, 10 ^-8 M or less than 10 ^-8 M, such as 10 ^-8 M to 10 ^-13 M, such as 10 ^-9 M to 10 ^-13 M).

In one example, Kd is measured by radiolabeled antigen binding analysis (RIA) using the Fab type of the relevant antibody and its antigen, as described in the following analysis. The solution binding affinity of Fab to antigen is achieved by balancing the Fab with a minimum concentration of ( ¹²⁵ I) labeled antigen in the presence of a titration series of unlabeled antigen, and then capturing the bound antigen with a plate coated with anti-Fab antibody. Test ( see, for example, Chen et al., J. Mol. Biol. 293:865-881 (1999)). In order to establish the assay conditions, MICROTITER ^® multi-well plates (Thermo Scientific) were coated with 50 mM sodium carbonate (pH 9.6) containing 5 µg/ml capture anti-Fab antibody (Cappel Labs) overnight, and then at room temperature (about 23%). Use PBS containing 2% (w/v) bovine serum albumin to block for two to five hours at ℃). In a non-absorbent disk (Nunc #269620), mix 100 pM or 26 pM [ ¹²⁵ I]-antigen with serial dilutions of related Fab ( for example, in Presta et al., Cancer Res. 57:4593-4599 (1997) The evaluation of anti-VEGF antibody Fab-12 is consistent). The Fab of interest is subsequently incubated overnight; however, the incubation can continue for a longer period of time (e.g. 65 hours) to ensure that equilibrium is reached. Thereafter, the mixture is transferred to a capture culture dish for incubation at room temperature (for example, for one hour). The solution was then removed and the dish was washed eight times with PBS containing 0.1% polysorbate 20 (TWEEN-20 ^® ). When the culture plate is dry, add 150 microliters/well of scintillator (MICROSCINT-20 ^™ ; Packard), and count the culture plate on a TOPCOUNT ^™ gamma counter (Packard) for 10 minutes. The concentration of each Fab that provides less than or equal to 20% of the maximum binding is selected for the competitive binding assay.

According to another embodiment, using BIACORE ^® -2000 or BIACORE ^® -3000 (BIAcore, Piscataway, NJ), using a fixed antigen CM5 chip at about 10 reaction units (RU) and using surface plasmon at 25°C Resonance analysis is used to measure Kd. In short, use N -ethyl- N' -(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N -hydroxysuccinimide (NHS) according to the supplier’s instructions. ) Activated carboxymethylated dextran biosensor chip (CM5, BIACORE company). The antigen was diluted to 5 μg/ml (about 0.2 μM) with 10 mM sodium acetate (pH 4.8), and then injected at a flow rate of 5 μl/min to achieve about 10 reaction units (RU) of coupled protein. After injection of antigen, 1 M ethanolamine was injected to block unreacted groups. For kinetic measurement, two-fold serial dilutions of Fab (0.78 nM to 500 nM) were injected into 0.05% polysorbate 20 (TWEEN-20 ^TM ) at a flow rate of about 25 μl/min at 25°C Surfactant (PBST) in PBS. By simultaneously fitting the binding and dissociation sensorgrams, a simple one-to-one Langmuir binding model (BIACORE ^® evaluation software version 3.2) is used to calculate the binding rate (k _on ) and dissociation rate (k _off ). The equilibrium dissociation constant (Kd) is calculated as the ratio k _off /k _on . See, for example , Chen et al., J. Mol. Biol. 293:865-881 (1999). If the association rate obtained from the above surface plasmon resonance analysis exceeds 10 ⁶ M ^-1 s ^-1 , the association rate can be determined by using the fluorescence quenching technique, which is measured in the spectrometer ( Such as stop-flow equipped with a spectrophotometer (Aviv Instruments) or 8000 series SLM-AMINCO ^TM spectrophotometer (ThermoSpectronic) with stirring cuvette in the presence of an increasing concentration of antigen measured in the presence of 25 ℃ in PBS (pH 7.2) Increase or decrease of the fluorescence emission intensity (excitation = 295 nm; emission = 340 nm, 16 nm bandpass) of the 20 nM anti-antigen antibody (Fab format). B. Antibody fragments

In certain embodiments, the antibodies used in the treatment methods provided herein ( eg , anti-CD79b antibodies or anti-CD20 antibodies) are antibody fragments. Antibody fragments include (but are not limited to) Fab, Fab', Fab'-SH, F(ab') ₂ , Fv and scFv fragments, and other fragments described below. For a review of certain antibody fragments, see Hudson et al . Nat. Med. 9:129-134 (2003). For a review of scFv, see Pluckthün, in The Pharmacology of Monoclonal Antibodies , Volume 113, Rosenburg and Moore eds, (Springer-Verlag, New York), pages 269-315 (1994); see also WO 93/16185; and US Patent Nos. 5,571,894 and 5,587,458. For a discussion of Fab and F(ab') ₂ fragments containing salvage receptor binding epitope residues and prolonged in vivo half-life, see US Patent No. 5,869,046.

Bifunctional antibodies are antibody fragments with two antigen binding sites, which can be bivalent or bispecific antibody fragments. See, for example, EP 404,097; WO 1993/01161; Hudson et al., Nat. Med. 9:129-134 (2003); and Hollinger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993). Trifunctional antibodies and tetrafunctional antibodies are also described in Hudson et al., Nat. Med. 9:129-134 (2003).

Single-domain antibodies are antibody fragments that comprise all or part of the heavy chain variable domain of an antibody or all or part of the light chain variable domain. In certain embodiments, the single domain antibody is a human single domain antibody (Domantis, Waltham, MA; see, for example, U.S. Patent No. 6,248,516 B1).

Antibody fragments can be prepared by various techniques, including but not limited to proteolytic digestion of intact antibodies and as described herein, produced by recombinant host cells ( e.g. , E. coli or phage). C. Chimeric and humanized antibodies

In certain embodiments, the antibodies ( eg , anti-CD79b antibodies or anti-CD20 antibodies) used in the treatment methods provided herein are chimeric antibodies. Certain chimeric antibodies are described in, for example, US Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA , 81:6851-6855 (1984)). In one example, a chimeric antibody includes a non-human variable region ( e.g. , a variable region derived from a mouse, rat, hamster, rabbit, or a non-human primate (such as monkey)) and a human constant region. In another example, a chimeric antibody is a "class-switched" antibody, where the class or subclass has changed from the class or subclass of the parent antibody. Chimeric antibodies include their antigen-binding fragments.

In certain embodiments, the chimeric antibody is a humanized antibody. Generally, non-human antibodies are humanized to reduce immunogenicity to humans while maintaining the specificity and affinity of the parental non-human antibodies. Generally, a humanized antibody comprises one or more variable domains, where HVR, such as CDR (or part thereof) is derived from a non-human antibody, and FR (or part thereof) is derived from a human antibody sequence. The humanized antibody optionally also contains at least a part of the human constant region. In some embodiments, some FR residues in the humanized antibody are substituted with corresponding residues from non-human antibodies (eg, antibodies from which HVR residues are derived), for example, to restore or increase antibody specificity or affinity.

Humanized antibodies and their preparation methods are reviewed in, for example, Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008), and are further described in, for example, Riechmann et al., Nature 332:323-329 (1988); Queen et al. , Proc. Nat'l Acad. Sci. USA 86:10029-10033 (1989); U.S. Patent Nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409; Kashmiri et al., Methods 36:25-34 (2005) (Describe SDR (a-CDR) grafting); Padlan, Mol. Immunol. 28:489-498 (1991) (description "surface remodeling");Dall'Acqua et al., Methods 36:43-60 (2005) (description " FR swap"); and Osbourn et al., Methods 36:61-68 (2005) and Klimka et al., Br. J. Cancer , 83:252-260 (2000) (describe the "guided selection" method of FR swap).

Human framework regions that can be used for humanization include but are not limited to: framework regions selected using the "best fit" method ( see, for example, Sims et al . J. Immunol. 151:2296 (1993)); derived from light chain or heavy chain The framework region of the consensus sequence of a human antibody of a specific subset of the variable region ( see, for example, Carter et al . Proc. Natl. Acad. Sci. USA , 89: 4285 (1992); and Presta et al . J. Immunol. , 151: 2623 (1993)); human mature (somatic mutation) framework regions or human germline framework regions ( see, for example, Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and obtained from screening FR libraries Framework regions ( see, for example, Baca et al., J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271:22611-22618 (1996)). D. Human antibodies

In certain embodiments, the antibodies used in the treatment methods provided herein (eg, anti-CD79b antibodies or anti-CD20 antibodies) are human antibodies. Human antibodies can be produced using various techniques known in the art. Human antibodies are generally described in van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5: 368-74 (2001) and Lonberg, Curr. Opin. Immunol. 20: 450-459 (2008).

Human antibodies can be prepared by administering immunogens to transgenic animals that have been engineered to produce intact human antibodies or intact antibodies with human variable regions in response to antigen challenge. Such animals usually contain all or part of the human immunoglobulin locus, which replaces the endogenous immunoglobulin locus, or exists outside the chromosomes or is randomly integrated into the animal chromosomes. In such transgenic mice, the endogenous immunoglobulin locus is generally inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23:1117-1125 (2005). See also, for example , U.S. Patent Nos. 6,075,181 and 6,150,584 describing XENOMOUSE ^TM technology; U.S. Patent No. 5,770,429 describing HuMab® technology; U.S. Patent No. 7,041,870 describing KM MOUSE® technology and U.S. Patent Application Publications describing VelociMouse® technology No. US 2007/0061900. The human variable regions of intact antibodies produced by such animals can be further modified, for example, by combining with different human constant regions.

Human antibodies can also be produced by fusion tumor-based methods. Human myeloma and mouse-human hybrid myeloma cell lines for the production of human monoclonal antibodies have been described. ( See, for example, Kozbor J. Immunol. , 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications , pp. 51-63 (Marcel Dekker, New York, 1987); and Boerner et al., J. Immunol. , 147: 86 (1991).) Human antibodies produced by human B-cell fusion tumor technology are also described in Li et al . Proc. Natl. Acad. Sci. USA , 103:3557-3562 (2006). Other methods include, for example, those described in the following documents: U.S. Patent No. 7,189,826 (describes the production of monoclonal human IgM antibodies from fusion tumor cell lines) and Ni, Xiandai Mianyixue , 26(4):265-268 (2006)( Describe human-human fusion tumors). Human fusion tumor technology (triple fusion tumor technology) is also described in Vollmers and Brandlein, Histology and Histopathology , 20(3):927-937 (2005) and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology , 27(3 ): 185-91 (2005).

Human antibodies can also be produced by isolating Fv cloned variable domain sequences selected from human phage display libraries. Such variable domain sequences can then be combined with the desired human constant domains. The techniques used to select human antibodies from antibody libraries are described below. E. Library source antibodies

In some embodiments, antibodies used in the treatment methods provided herein (for example, anti-CD79b antibodies or anti-CD20 antibodies) can be isolated by screening combinatorial libraries to obtain antibodies with the desired activity. For example, various methods are known in the art for generating phage display libraries and screening such libraries for antibodies with desired binding characteristics. These methods are reviewed in, for example, Hoogenboom et al., Methods in Molecular Biology 178:1-37 (O'Brien et al., eds., Human Press, Totowa, NJ, 2001), and are further described in, for example, McCafferty et al., Nature 348: 552-554; Clackson et al., Nature 352:624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Marks and Bradbury, in Methods in Molecular Biology 248:161- 175 (Editor Lo, Human Press, Totowa, NJ, 2003); Sidhu et al., J. Mol. Biol. 338(2):299-310 (2004); Lee et al., J. Mol. Biol. 340(5 ): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2): 119 -132 (2004).

In some phage display methods, the lineages of VH and VL genes are respectively selected by polymerase chain reaction (PCR) and randomly recombined into phage libraries, and then the antigen-binding phages in these libraries can be screened, such as Winter, etc. Human Ann. Rev. Immunol. , 12: 433-455 (1994). Phages usually display antibody fragments, either as single chain Fv (scFv) fragments or as Fab fragments. Libraries from immune sources provide high-affinity antibodies against immunogens without the need to construct fusion tumors. Alternatively, the natural lineage can be cloned (eg selected from humans) to provide a single source of antibodies against a wide range of non-self antigens and self-antigens without any immunization, such as Griffiths et al. EMBO J , 12: 725- 734 (1993). Finally, natural libraries can also be prepared synthetically by the following methods: unrearranged V gene segments are cloned from stem cells, and PCR primers containing random sequences are used to encode highly variable CDR3 regions and rearrangement in vitro, such as Hoogenboom and Winter, J. Mol. Biol. , 227: 381-388 (1992). Patent publications describing human antibody phage libraries include, for example, U.S. Patent No. 5,750,373, and U.S. Patent Publication Nos. 2005/0079574, 2005/0119455, 2005/0266000, 2007/0117126, 2007/0160598, 2007/0237764, 2007 /0292936 and 2009/0002360.

Antibodies or antibody fragments isolated from human antibody libraries are referred to herein as human antibodies or human antibody fragments. F. Multispecific antibodies

In certain embodiments, the antibodies (eg, anti-CD79b antibodies or anti-CD20 antibodies) used in the treatment methods provided herein are multispecific antibodies, such as bispecific antibodies. Multispecific antibodies are monoclonal antibodies that have binding specificities for at least two different sites. In certain embodiments, one of the binding specificities is for one antigen ( eg, CD79b or CD20), and the other binding specificity is for any other antigen. In certain embodiments, one binding specificity is for one antigen ( eg CD79b or CD20) and the other is for CD3. See, for example, U.S. Patent No. 5,821,337. In certain embodiments, bispecific antibodies can bind to two different epitopes of a single antigen ( e.g. , CD79b or CD20). Bispecific antibodies can also be used to localize cytotoxic agents to cells expressing antigens ( eg , CD79b or CD20). Bispecific antibodies can be prepared as full-length antibodies or antibody fragments.

Techniques for preparing multispecific antibodies include (but are not limited to) recombinant co-expression of two immunoglobulin heavy chain-light chain pairs with different specificities ( see Milstein and Cuello, Nature 305: 537 (1983)), WO 93/ 08829 and Traunecker et al. EMBO J. 10: 3655 (1991)) and "knob-in-hole" engineering ( see, for example, U.S. Patent No. 5,731,168). Multispecific antibodies can also be prepared by the following methods: the electrostatic steering effect designed to prepare antibody Fc-heterodimer molecules (WO 2009/089004A1); cross-linking two or more antibodies or fragments ( see for example U.S. Patent No. 4,676,980, and Brennan et al., Science , 229: 81 (1985); use leucine zip to produce bispecific antibodies ( see, for example, Kostelny et al., J. Immunol. , 148(5): 1547 -1553 (1992)); using "mini-bifunctional antibody" technology to prepare bispecific antibody fragments ( see, for example, Hollinger et al., Proc. Natl. Acad. Sci. USA , 90:6444-6448 (1993)); and use Single-chain Fv (sFv) dimers ( see, for example, Gruber et al., J. Immunol. , 152:5368 (1994)); and as described, for example, in Tutt et al . J. Immunol. 147: 60 (1991) Preparation of trispecific antibodies.

Also included herein are engineered antibodies with three or more functional antigen binding sites, including "Octopus antibodies" ( see, for example, US 2006/0025576A1).

The antibody or fragment herein also includes "dual-acting FAb" or "DAF", which includes an antigen binding site that binds to CD79b and another different antigen ( see, for example, US 2008/0069820). G. Antibody variants

In certain embodiments, amino acid sequence variants of the antibodies used in the treatment methods provided herein ( eg , anti-CD79b antibodies or anti-CD20 antibodies) are covered. For example, it may be necessary to improve the binding affinity and/or other biological properties of an anti-CD79b antibody or an anti-CD20 antibody. The amino acid sequence variants of the antibody can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletion and/or insertion and/or substitution of residues within the amino acid sequence of the antibody. Any combination of deletion, insertion, and substitution can be performed to obtain the final construct, and the limitation is that the final construct has the required characteristics, such as antigen binding. (i) Substitution, insertion and deletion variants

In certain embodiments, antibody variants with one or more amino acid substitutions are provided. The sites of interest for substitution mutagenesis include HVR and FR. Conservative substitutions are shown in Table D under the heading "Preferred Substitutions". More substantive changes are provided under the heading "Exemplary Substitutions" in Table D and are further described below with reference to amino acid side chain classes. Amino acid substitutions can be introduced into the antibody of interest, and the product screened for desired activities such as retained/improved antigen binding, reduced immunogenicity, or improved ADCC or CDC. Table D

Amino acids can be grouped according to common side chain characteristics: (1) Hydrophobicity: Leucine, Met, Ala, Val, Leu, Ile; (2) Neutral hydrophilicity: Cys, Ser, Thr, Asn, Gln; (3) Acidity: Asp, Glu; (4) Basicity: His, Lys, Arg; (5) Residues that affect chain orientation: Gly, Pro; (6) Aromaticity: Trp, Tyr, Phe.

Non-conservative substitutions will require replacing a member of one of these categories with another.

One type of substitution variant involves the substitution of one or more hypervariable region residues of a parent antibody ( e.g. , a humanized or human antibody). Generally, the resulting variants that are selected for further research will have certain biological characteristic modifications ( e.g. , improvements) ( e.g. , increased affinity, decreased immunogenicity) and/or relative to the parent antibody It will have certain biological properties of the parent antibody that are substantially maintained. An exemplary substitution variant is an affinity maturation antibody, which can be conveniently produced, for example, using affinity maturation techniques based on phage presentation, such as those described herein. In short, one or more HVR residues are mutated and the variant antibodies are displayed on phage and screened for specific biological activity ( e.g. , binding affinity).

Changes ( e.g. , substitutions) can be made in HVR, for example to improve antibody affinity. Can be in HVR ``hot spots'' (i.e. residues encoded by codons that undergo mutations at a high frequency during the maturation process of somatic cells) ( see, for example, Chowdhury, Methods Mol. Biol. 207:179-196 (2008)) and/ Or make these changes in SDR (a-CDR), and test the binding affinity of the obtained variant VH or VL. For example, affinity maturation by construction and reselection from secondary libraries is described in the following literature: Hoogenboom et al., Methods in Molecular Biology 178:1-37 (O'Brien et al. eds., Human Press, Totowa, NJ, (2001).) In some embodiments of affinity maturation, diversity is introduced into any one selected for use by a variety of methods ( e.g. , error-prone PCR, strand shuffling, or oligonucleotide-directed mutagenesis) In the variable genes that achieve maturity. Then a second library is generated. The library is then screened to identify any antibody variants with the desired affinity. Another method of introducing diversity involves HVR site-directed methods, in which several HVR residues ( for example , 4-6 residues at a time) are randomized. HVR residues involved in antigen binding can be specifically identified, for example, using alanine scanning mutagenesis or modeling. Usually CDR-H3 and CDR-L3 are especially targeted.

In certain embodiments, substitutions, insertions, or deletions may occur in one or more HVRs, as long as the changes do not substantially reduce the ability of the antibody to bind antigen. For example, conservative changes ( e.g. , conservative substitutions as provided herein) can be made in HVR that do not substantially reduce binding affinity. These changes can be outside the HVR "hot spot" or SDR. In certain embodiments of the variant VH and VL sequences provided above, each HVR is unchanged or contains only one, two or three amino acid substitutions.

A suitable method for identifying residues or regions in antibodies that can be targeted for mutagenesis is called "alanine scanning mutagenesis", as described by Cunningham and Wells (1989) Science , 244:1081-1085. In this method, a certain residue or a set of target residues ( e.g., charged residues such as arg, asp, his, lys, and glu) are recognized and replaced with a neutral or negatively charged amino acid ( e.g., alanine Or polyalanine) to determine whether the interaction between antibody and antigen is affected. Further substitution can be introduced at the amino acid position, demonstrating the functional sensitivity to the initial substitution. Alternatively or additionally, the crystal structure of the antigen-antibody complex is used to identify contact points between the antibody and the antigen. These contact residues and neighboring residues can be targeted or eliminated as candidates for substitution. The variants can be screened to determine whether they contain the desired characteristics.

Amino acid sequence insertions include amino- and/or carboxy-terminal fusions ranging from one residue to the length of polypeptides containing one hundred or more residues, and sequences of single or multiple amino acid residues内 Insert. Examples of terminal insertions include antibodies with N-terminal methionine residues. Other insertion variants of the antibody molecule include the N-terminus or C-terminus of the antibody fused to an enzyme ( for example , for ADEPT) or a polypeptide that increases the serum half-life of the antibody. (ii) Glycosylation variants

In certain embodiments, the antibodies used in the treatment methods provided herein ( eg , anti-CD79b antibodies or anti-CD20 antibodies) are changed to increase or decrease the degree of antibody glycosylation. The addition of glycosylation sites to the antibody or the deletion of glycosylation sites in the antibody can be conveniently achieved by changing the amino acid sequence to create or remove one or more glycosylation sites.

In the case where the antibody contains an Fc region, the carbohydrate attached to it can be changed. Native antibodies produced by mammalian cells typically contain branched, biantennary oligosaccharides, which are generally linked to Asn297 in the CH2 domain of the Fc region by N-linking. See, for example, Wright et al. TIBTECH 15:26-32 (1997). The oligosaccharide may include a variety of carbohydrates, such as mannose, N-acetylglucosamine (GlcNAc), galactose and sialic acid, and seaweed linked to GlcNAc in the "stem" of the biantennary oligosaccharide structure sugar. In some embodiments, modification of the oligosaccharide in the antibody of the invention can be performed to produce antibody variants with certain improved properties.

In one embodiment, antibody variants having a carbohydrate structure lacking (directly or indirectly) trehalose linked to the Fc region are provided. For example, the amount of trehalose in the antibody can be 1% to 80%, 1% to 65%, 5% to 65%, or 20% to 40%. The amount of trehalose is calculated by calculating the trehalose at Asn297 in the sugar chain relative to all sugar structures attached to Asn 297 as measured by MALDI-TOF mass spectrometry analysis (e.g., complex, hybrid and high mannose structures) The sum is determined by the average amount, as described in, for example, WO 2008/077546. Asn297 refers to the asparagine residue located at approximately position 297 in the Fc region (Eu numbering of residues in the Fc region); however, Asn297 can also be located approximately ±±± position upstream or downstream of position 297 due to minor sequence changes in the antibody 3 amino acids, that is, between positions 294 and 300. These trehalcosylation variants may have improved ADCC function. See, for example, US Patent Publication US 2003/0157108 (Presta, L.); US 2004/0093621 (Kyowa Hakko Kogyo Co., Ltd). Examples of publications related to "de-trehaloseylation" or "trehalose deficiency" antibody variants include: US 2003/0157108; WO 2000/61739; WO 2001/29246; US 2003/0115614; US 2002/0164328; US 2004/0093621; US 2004/0132140; US 2004/0110704; US 2004/0110282; US 2004/0109865; WO 2003/085119; WO 2003/084570; WO 2005/035586; WO 2005/035778; WO2005/053742; WO2002 /031140; Okazaki et al. J. Mol. Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al. Biotech. Bioeng. 87:614 (2004). Examples of cell lines capable of producing anti-trehaloseylated antibodies include Lec13 CHO cells lacking protein trehalosylation (Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); U.S. Patent Application No. US 2003/0157108 A1, Presta, L; and WO 2004/056312 A1, Adams et al ., especially Example 11) and gene knockout cell lines, such as α-1,6-trehalosyltransferase gene FUT8 gene knockout CHO cells ( See, for example, Yamane-Ohnuki et al. Biotech. Bioeng. 87:614 (2004); Kanda, Y. et al., Biotechnol. Bioeng. , 94(4):680-688 (2006); and WO2003/085107).

The antibody variant further has a bisected oligosaccharide, for example, the biantennary oligosaccharide attached to the Fc region of the antibody is divided equally by GlcNAc. These antibody variants may have reduced trehalosylation and/or improved ADCC function. Examples of such antibody variants are described in, for example, WO 2003/011878 (Jean-Mairet et al.); US Patent No. 6,602,684 (Umana et al.); and US 2005/0123546 (Umana et al. ). Also provided are antibody variants having at least one galactose residue in the oligosaccharide linked to the Fc region. These antibody variants may have improved CDC function. Such antibody variants are described in, for example, WO 1997/30087 (Patel et al.); WO 1998/58964 (Raju, S.); and WO 1999/22764 (Raju, S.). (iii) Fc variant

In certain embodiments, one or more amino acid modifications can be introduced into the Fc region of an antibody used in the treatment methods provided herein ( eg , an anti-CD79b antibody or an anti-CD20 antibody) to generate Fc region variants. The Fc region variant may comprise a human Fc region sequence ( e.g. , a human IgG1, IgG2, IgG3, or IgG4 Fc region) that includes an amino acid modification ( e.g. , substitution) at one or more amino acid positions.

In certain embodiments, the present invention covers antibody variants with some but not all effector functions. These effector functions make the antibody variants have important in vivo half-lives, but certain effector functions (such as complement And ADCC) candidate for unnecessary or harmful applications. In vitro and/or in vivo cytotoxicity analysis can be performed to confirm the reduction/exhaustion of CDC and/or ADCC activity. For example, an Fc receptor (FcR) binding analysis can be performed to ensure that the antibody lacks FcγR binding (and therefore may lack ADCC activity), but retains FcRn binding ability. The primary cells used to mediate ADCC, NK cells, only express Fc (RIII, while monocytes express Fc (RI, Fc(RII, and Fc(RIII). The expression of FcR on hematopoietic cells is summarized in Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991) page 464 in Table 3. A non-limiting example of an in vitro assay for assessing the ADCC activity of the molecule of interest is described in U.S. Patent No. 5,500,362 ( see, for example, Hellstrom, I . Et al. Proc. Nat ' l Acad. Sci. USA 83:7059-7063 (1986)) and Hellstrom, I et al . Proc. Nat ' l Acad. Sci. USA 82:1499-1502 (1985); 5,821,337 ( see Bruggemann, M. et al., J. Exp. Med. 166:1351-1361 (1987)). Alternatively, a non-radioactive assay method can be used ( see, for example, the ACTI™ non-radioactive cytotoxicity assay of flow cytometry (CellTechnology). Company. Mountain View, CA); and CytoTox 96 ^® non-radioactive cytotoxicity assay (Promega, Madison, WI). Effector cells suitable for these analyses include peripheral blood mononuclear cells (PBMC) and natural killer (NK) Cells. Alternatively or additionally, the ADCC activity of related molecules can be evaluated in vivo, for example, in animal models, such as those disclosed in Clynes et al . Proc. Nat'l Acad. Sci. USA 95:652-656 (1998). C1q binding analysis can also be performed to confirm that the antibody cannot bind to C1q and therefore lacks CDC activity. See, for example , the C1q and C3c binding ELISA in WO 2006/029879 and WO 2005/100402. To assess complement activation, CDC assays can be performed ( see for example Gazzano-Santoro et al ., J. Immunol. Methods 202:163 (1996); Cragg, MS et al., Blood 101:1045-1052 (2003); and Cragg, MS and MJ Glennie, Blood 103:2738-2743 (2004) )). Methods known in the art can also be used to determine FcRn binding and in vivo clearance/half-life ( see, for example, Petkova, SB et al., Int'l . Immunol. 18(12): 1759-1769 (2006) )) .

Antibodies with reduced effector functions include those having substitutions of one or more of Fc region residues 238, 265, 269, 270, 297, 327, and 329 (US Patent No. 6,737,056). Such Fc mutants include Fc mutants with substitutions of two or more of amino acid positions 265, 269, 270, 297 and 327, including the so-called "DANA" Fc in which residues 265 and 297 are substituted with alanine Mutant (US Patent No. 7,332,581).

Certain antibody variants with improved or reduced binding to FcR are described. ( See, for example, U.S. Patent No. 6,737,056; WO 2004/056312, and Shields et al., J. Biol. Chem. 9(2): 6591-6604 (2001).)

In certain embodiments, antibody variants comprise an Fc region with one or more amino acid substitutions that improve ADCC, such as substitutions at positions 298, 333, and/or 334 (EU numbering of residues) in the Fc region.

In some embodiments, changes that result in C1q binding and/or complement dependent cytotoxicity (CDC) changes (ie, improvement or weakening) are performed in the Fc region, for example, as in US Patent No. 6,194,551, WO 99/51642 and Idusogie Et al. J. Immunol. 164: 4178-4184 (2000).

Antibodies with increased half-life and improved binding to the neonatal Fc receptor (FcRn) are described in US2005/0014934A1 (Hinton et al.). The neonatal Fc receptor (FcRn) is responsible for the transfer of maternal IgG to the fetus ( Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994)). These antibodies comprise an Fc region with one or more substitutions therein, and these substitutions improve the binding of the Fc region to FcRn. These Fc variants include those with substitutions in one or more of the Fc region residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424, or 434, such as a substitution at residue 434 in the Fc region (US Patent No. 7,371,826).

See also Duncan and Winter, Nature 322:738-40 (1988); U.S. Patent No. 5,648,260; U.S. Patent No. 5,624,821; and WO 94/29351, which relates to other examples of Fc region variants. (iv) Antibody variants modified by cysteine

In some embodiments, it may be necessary to generate cysteine-engineered antibodies, such as "thioMAb", in which one or more residues of the anti-CD79b antibody or anti-CD20 antibody used in the treatment methods provided herein are cysteamine Acid residue substitution. In a specific embodiment, the substituted residue is present at an accessible site of the antibody. By substituting cysteine for these residues, the reactive thiol group is thus positioned at the accessible site of the antibody and can be used to bind the antibody to other parts (such as the drug moiety or the linker-drug moiety) ) To produce immunoconjugates, as described further herein. In certain embodiments, any one or more of the following residues can be substituted with cysteine: V205 (Kabat numbering) for the light chain; A118 for the heavy chain (EU numbering); and for the Fc region of the heavy chain S400 (EU number). Cysteine-engineered antibodies can be produced as described in, for example, US Patent No. 7,521,541. (v) Antibody derivatives

In certain embodiments, the antibodies used in the treatment methods provided herein ( eg , anti-CD79b antibodies or anti-CD20 antibodies) can be further modified to include other non-protein moieties known and readily available in the art. Parts suitable for derivatization of antibodies include but are not limited to water-soluble polymers. Non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), ethylene glycol/propylene glycol copolymer, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly -1,3-dioxolane, poly-1,3,6-trioxane, ethylene/maleic anhydride copolymer, polyamino acid (homopolymer or random copolymer) and dextran Or poly(n-vinylpyrrolidone) polyethylene glycol, polypropylene glycol homopolymer, polypropylene oxide/ethylene oxide copolymer, polyoxyethylated polyol ( for example , glycerin), polyvinyl alcohol and mixtures thereof . Polyethylene glycol propionaldehyde may have advantages in manufacturing due to its stability in water. The polymer can have any molecular weight, and can be branched or unbranched. The number of polymers attached to the antibody can vary, and if more than one polymer is attached, they can be the same or different molecules. Generally, the number and/or type of polymers used for derivatization can be determined based on a variety of considerations, including but not limited to the specific characteristics or functions of the antibody to be improved, and whether the antibody derivative will be under specified conditions. It is used for therapy and so on.

In another embodiment, a conjugate of an antibody and a non-protein moiety that can be selectively heated by exposure to radiation is provided. In one embodiment, the non-protein portion is a carbon nanotube (Kam et al., Proc. Natl. Acad. Sci. USA 102: 11600-11605 (2005)). The radiation can have any wavelength and includes (but is not limited to) wavelengths that do not damage normal cells but heat the non-protein part to a temperature that kills the cells near the antibody-non-protein part. H. Recombination method and composition

Antibodies can be produced using, for example, recombinant methods and compositions as described in U.S. Patent No. 4,816,567. In one embodiment, an isolated nucleic acid encoding the antibody described herein is provided. Such nucleic acids may encode the amino acid sequence of the VL of an antibody and/or the amino acid sequence of the VH of the antibody ( e.g., the light chain and/or heavy chain of an antibody). In another embodiment, one or more vectors ( e.g., expression vectors) containing such nucleic acids are provided. In another embodiment, a host cell containing such nucleic acid is provided. In one such embodiment, the host cell comprises ( for example, the following transformation has been used): (1) a vector comprising a nucleic acid encoding a nucleic acid sequence comprising the amino acid sequence of the VL of the antibody and the amino acid sequence of the VH of the antibody, or (2 ) A first vector comprising a nucleic acid encoding the amino acid sequence of the VL of the antibody and a second vector comprising the nucleic acid encoding the amino acid sequence of the VH of the antibody. In one embodiment, the host cell is eukaryotic, such as Chinese Hamster Ovary (CHO) cells or lymphoid cells ( e.g., Y0, NS0, Sp20 cells). In one embodiment, there is provided a method for preparing an antibody, wherein the method comprises culturing a host cell comprising the nucleic acid encoding the antibody as provided above under conditions suitable for expressing the antibody, and optionally from the host cell (or Host cell culture medium) to recover the antibody.

For the recombinant production of antibodies, nucleic acids encoding antibodies such as those described above are isolated, and inserted into one or more vectors for further selection and/or expression in host cells. Such nucleic acids can be easily isolated and sequenced using conventional procedures ( for example , by using oligonucleotide probes that can specifically bind to genes encoding antibody heavy and light chains).

Host cells suitable for selection or expression of antibody-encoding vectors include the prokaryotic or eukaryotic cells described herein. For example, antibodies can be produced in bacteria, especially when glycosylation and Fc effector functions are not required. For the expression of antibody fragments and polypeptides in bacteria, see, for example, U.S. Patent Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (BKC Lo ed, Humana Press, Totowa, NJ, 2003), pp. 245-254, which describes the expression of antibody fragments in E. coli.) After the performance, can be The antibody is separated from the bacterial cell paste in the soluble fraction and can be further purified.

In addition to prokaryotes, eukaryotic microorganisms such as filamentous fungi or yeasts are suitable for selection or expression hosts for antibody-encoding vectors, including glycosylation pathways that have been "humanized" so that the antibodies produced have some Or fungi and yeast strains that are fully human glycosylated. See Gerngross, Nat. Biotech. 22:1409-1414 (2004) and Li et al., Nat. Biotech. 24:210-215 (2006).

Suitable host cells for expressing glycosylated antibodies are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. We have identified many be used in combination with insect cells, particularly for transfection of Spodoptera armyworm (Spodoptera frugiperda) cell line baculovirus.

Plant cell cultures can also be used as hosts. See, for example, U.S. Patent Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (description of PLANTIBODIES ^™ technology for the production of antibodies in transgenic plants).

Vertebrate cells can also be used as hosts. For example, mammalian cell strains suitable for growth in suspension may be suitable. Other examples of useful mammalian host cell strains are monkey kidney CV1 strains transformed by SV40 (COS-7); human embryonic kidney strains (as described, for example, in Graham et al., J. Gen Virol. 36:59 (1977) Described 293 or 293 cells); Hamster kidney cells (BHK); Mouse Sergio cells (such as, for example , TM4 cells described in Mather, Biol. Reprod. 23:243-251 (1980)); Monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical cancer cells (HELA); canine kidney cells (MDCK; Buffalo rat liver cells (BRL 3A); human lung cells (W138); human liver Cells (Hep G2); Mouse breast tumors (MMT 060562); TRI cells, as described, for example, in Mather et al., Annals NY Acad. Sci. 383:44-68 (1982); MRC 5 cells; and FS4 cells Other suitable mammalian host cell lines include Chinese Hamster Ovary (CHO) cells, including DHFR ^- CHO cells (Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)); and myeloma cell lines, such as Y0, NS0 and Sp2 / 0. for a review of certain mammalian host cell lines suitable for producing the antibody, see, e.g., Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (BKC Lo ed, Humana Press, Totowa, NJ ), pages 255-268 (2003). I. Verification

The antibodies used in the treatment methods provided herein (for example, anti-CD79b antibodies or anti-CD20 antibodies) can be identified, screened or characterized by various assays known in the art for their physical/chemical properties and/or biological activities.

In one aspect, a method of treating the test provided herein, antibodies (e.g., anti-CD79b antibody or anti-CD20 antibody) The antigen binding activity, for example by known methods such as ELISA, BIACore ^®, FACS or Western blotting.

In another aspect, competition analysis can be used to identify antibodies that compete with any of the antibodies described herein for binding to the target antigen. In certain embodiments, the competitive antibody binds to the same epitope ( e.g., linear or conformational epitope) bound by the antibodies described herein. A detailed exemplary method for locating the epitope bound by the antibody is provided in Morris (1996) "Epitope Mapping Protocols", in Methods in Molecular Biology, Volume 66 (Humana Press, Totowa, NJ).

In an exemplary competition analysis, the immobilized antigen is incubated in a solution containing a first labeled antibody that binds to the antigen ( such as any antibody described herein) and a second unlabeled antibody, the second unlabeled antibody and the first The ability of an antibody to compete with antigen is tested. The second antibody may be present in the supernatant of the fusion tumor. As a control group, the immobilized antigen was incubated in a solution containing the first labeled antibody but no second unlabeled antibody. After incubation under conditions that allow the first antibody to bind to the antigen, excess unbound antibody is removed, and the amount of label associated with the immobilized antigen is measured. If the amount of label associated with the immobilized antigen is substantially reduced in the test sample relative to the control sample, it indicates that the second antibody competes with the first antibody for binding to the antigen. See Harlow and Lane (1988) Antibodies: A Laboratory Manual Chapter 14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY). VIII. Pharmaceutical formulations

The pharmaceutical formulations of any of the agents described herein ( for example , anti-CD79b immunoconjugates, anti-CD20 agents, and alkylating agents) used in any of the methods described herein are obtained by combining an agent with the required purity with one or more optional The pharmaceutically acceptable carrier ( Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)) is mixed to prepare a lyophilized formulation or an aqueous solution. Pharmaceutically acceptable carriers are non-toxic to the recipient at the dose and concentration used, and include but are not limited to: buffers, such as phosphate, citrate and other organic acids; antioxidants, including ascorbic acid and methionine ; Preservatives (such as octadecyl dimethyl benzyl ammonium chloride; hexaalkyl quaternary ammonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl or benzyl alcohol; p-hydroxybenzoic acid Alkyl esters, such as methyl or propyl p-hydroxybenzoate; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) Polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulin; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, histidine, Arginine or lysine; monosaccharides, disaccharides and other carbohydrates, including glucose, mannose, or dextrin; chelating agents, such as EDTA; sugars, such as sucrose, mannitol, trehalose or sorbitol; Salt-forming relative ions, such as sodium; metal complexes (for example, Zn-protein complexes); and/or nonionic surfactants, such as polyethylene glycol (PEG). The exemplary pharmaceutically acceptable carriers herein further include interstitial drug dispersants, such as soluble neutral active hyaluronidase glycoprotein (sHASEGP), for example, human soluble PH-20 hyaluronidase glycoprotein, such as rHuPH20 (HYLENEX ^® , Baxter International). Some exemplary sHASEGP (including rHuPH20) and methods of use are described in U.S. Patent Publication Nos. 2005/0260186 and 2006/0104968. In one aspect, sHASEGP is combined with one or more additional mucopolysaccharidase (such as chondroitinase).

Exemplary freeze-dried antibody or immunoconjugate formulations are described in U.S. Patent No. 6,267,958. Aqueous antibody or immunoconjugate formulations include those described in U.S. Patent No. 6,171,586 and WO2006/044908, the latter formulations including histidine-acetate buffer.

The formulation herein may also contain more than one active ingredient as necessary for the specific indication being treated, preferably an active ingredient with complementary activities that does not adversely affect each other.

The active ingredients can be embedded in microcapsules prepared, for example, by coacervation technology or by interfacial polymerization, such as in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and Nanocapsules) or hydroxymethylcellulose or gelatin-microcapsules and poly-(methyl methacrylate) microcapsules in macroemulsion. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980).

Sustained release formulations can be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing antibodies or immunoconjugates, which matrices are in the form of shaped articles, such as films or microcapsules.

The formulations intended for in vivo administration are generally sterile. Sterility can be easily achieved by, for example, filtration through a sterile filter membrane.

Additional details about the pharmaceutical formulations containing anti-CD79 immunoconjugates are provided in WO 2009/099728, the content of which is expressly incorporated herein by reference in its entirety. IX. Manufacturing objects and sets

, 其中Ab為抗CD79b抗體，其包含(i)包含胺基酸序列SEQ ID NO: 21之HVR-H1，(ii)包含胺基酸序列SEQ ID NO: 22之HVR-H2，(iii)包含胺基酸序列SEQ ID NO: 23之HVR-H3，(iv)包含胺基酸序列SEQ ID NO: 24之HVR-L1，(v)包含胺基酸序列SEQ ID NO: 25之HVR-L2，及(vi)包含胺基酸序列SEQ ID NO: 26之HVR-L3；且其中p在1至8之範圍內。在一些實施例中，套組包含含有下式之免疫結合物

, 其中Ab係抗CD79b抗體，其包含(i)包含VH之重鏈，該VH包含SEQ ID NO：19之胺基酸序列，及(ii)包含VL之輕鏈，該VL包含SEQ ID NO：20之胺基酸序列，其中p在2與5之間。在一些實施例中，p介於3與4之間，例如3.5。在一些實施例中，免疫結合物包含抗CD79抗體，其包含含有SEQ ID NO：36之胺基酸序列之重鏈，並且其中輕鏈包含SEQ ID NO：35之胺基酸序列。在某些實施例中，抗CD79b免疫結合物包含Ab-MC-vc-PAB-MMAE之結構。在一些實施例中，抗CD79b免疫結合物係polatuzumab vedotin (CAS號1313206-42-6)。在一些實施例中，該至少一種另外藥劑係烷化劑(例如苯達莫司汀，例如苯達莫司汀-HCl)及抗CD20抗體(例如利妥昔單抗)。In another embodiment, an article or kit is provided that includes an anti-CD79b immunoconjugate (as described herein) and at least one additional agent. In some embodiments, the at least one additional agent is an alkylating agent (such as bendamustine, such as bendamustine-HCl) and an anti-CD20 antibody (such as rituximab). In some embodiments, the article or kit further comprises a package insert comprising combining the anti-CD79b immunoconjugate with at least one additional agent such as an alkylating agent (e.g., bendamustine, such as bendamol) Instructions for use of Stine-HCl) and an anti-CD20 antibody (e.g., rituximab) together to treat or delay the progression of a B cell proliferative disease (e.g., DLBCL) in an individual. Any anti-CD79b immunoconjugates and anticancer agents known in the art can be included in the product or kit. In some embodiments, the kit includes an immunoconjugate containing the formula

, Where Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22, and (iii) comprising The amino acid sequence of HVR-H3 of SEQ ID NO: 23, (iv) the HVR-L1 of the amino acid sequence of SEQ ID NO: 24, (v) the HVR-L2 of the amino acid sequence of SEQ ID NO: 25, And (vi) HVR-L3 comprising the amino acid sequence SEQ ID NO: 26; and wherein p is in the range of 1 to 8. In some embodiments, the kit includes an immunoconjugate containing the formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) a heavy chain comprising VH, which VH comprises the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, which VL comprises SEQ ID NO: The amino acid sequence of 20, where p is between 2 and 5. In some embodiments, p is between 3 and 4, such as 3.5. In some embodiments, the immunoconjugate comprises an anti-CD79 antibody, which comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 36, and wherein the light chain comprises the amino acid sequence of SEQ ID NO: 35. In certain embodiments, the anti-CD79b immunoconjugate comprises the structure of Ab-MC-vc-PAB-MMAE. In some embodiments, the anti-CD79b immunoconjugate is polatuzumab vedotin (CAS number 1313206-42-6). In some embodiments, the at least one additional agent is an alkylating agent (such as bendamustine, such as bendamustine-HCl) and an anti-CD20 antibody (such as rituximab).

In some embodiments, the kit is used to treat DLBCL in an individual (eg, an individual having one or more of the characteristics described herein) according to the methods provided herein.

In some embodiments, the anti-CD79 immunoconjugate, alkylating agent (such as bendamustine, such as bendamustine-HCl) and anti-CD20 antibody (such as rituximab) are in the same container or in separate In the container. Suitable containers include, for example, bottles, vials, bags, syringes, and the like. The container may be formed of various materials, such as glass, plastic (such as polyvinyl chloride or polyolefin), or metal alloy (such as stainless steel or Hastelloy). In some embodiments, the container contains the formulation, and a label on or associated with the container may indicate instructions for use. The article or kit may further include other materials desired from a commercial and user point of view, including other buffers, diluents, filters, needles, syringes, and package inserts with instructions for use. In some embodiments, the product further includes one or more other agents (such as chemotherapeutic agents and anti-tumor agents). Suitable containers for one or more medicaments include, for example, bottles, vials, bags, and syringes.

It is believed that the foregoing written description is sufficient to enable those familiar with the art to practice the present invention. In addition to the examples shown and described in this article, those who are familiar with the art will clearly understand the various modifications of the present invention based on the foregoing description, and they fall within the scope of the attached patent application. All publications, patents and patent applications cited herein are incorporated by reference for all purposes. Instance

The following are examples of the method and composition of the present invention. It should be understood that given the general description provided above, many other embodiments can be practiced. Example 1 : Anti- CD79b immunoconjugate (Polatuzumab Vedotin), anti- CD20 antibody ( rituximab ) and alkylating agent ( bendamustine ) are used in combination with relapsed or refractory diffuse large B- cell lymphoma (DLBCL) )

CD79b is located in normal B-cells and most mature B-cell malignant tumors, including >95% of the signal component of the B-cell receptor on DLBCL (Dornan et al., Blood, 114:2721-9, 2009; Pfeifer et al., Leukemia, 29:1578-86, 2015). Polatuzumab vedotin (CAS No. 1313206-42-6) has shown encouraging activity in R/R DLBCL as a monotherapy (Palanca-Wessels et al., Lancet Oncology, 16:704-15, 2015)). The CD20 monoclonal antibody combination (Morschhauser et al., Journal of Clinical Oncology, 32:15_suppl, 8519, 2014) produced an overall response rate (ORR) between 13-56%. However, the complete response (CR) rate is very low (0-15%), prompting the use of combination with other drugs. Bendamustine and rituximab (BR) are commonly used in R/R DLBCL that is not suitable for transplantation, and the reported median progression-free survival (PFS) is 3.6-6.7 months (Ohmachi et al., Journal of Clinical Oncology) 31:2103-9, 2013; Vacirca et al., Annals of Hematology, 93:403-9, 2014). The following describes the results of the Phase Ib/II trial, which evaluated polatuzumab vedotin, bendamustine, and atorizumab in R/R DLBCL that are not suitable for transplantation, including patients who have failed previous autologous stem cell transplantation (ASCT) The combination of antibodies (Pola-BG) and the combination of polatuzumab vedotin and BR (Pola-BR) were compared with BR alone. patient

Patients aged 18 years or older who have relapsed/refractory diffuse large B-cell lymphoma (R/R DLBCL) confirmed by biopsy after ≥1 previous treatment, Eastern Cooperative Oncology Group (ECOG) physical status If it is 0-2, peripheral neuropathy (PN) ≤ 1 grade, it is suitable for inclusion in this study. Suitable patients are deemed unsuitable for autologous stem cell transplantation (SCT) by the treating physician, or the previous SCT has failed. If SCT occurs> 100 days before the first day of treatment cycle 1, patients with a history of SCT are eligible.

Patients with grade 3b follicular lymphoma, transforming lymphoma, transforming and mild non-Hodgkin lymphoma (NHL) and central nervous system (CNS) lymphoma were excluded. Patients suitable for SCT were excluded. Patients who had received previous allogeneic stem cell transplantation were excluded. Test design

The Phase Ib safety trial included 6 patients treated with polatuzumab vedotin combined with bendamustine and rituximab ("Pola-BR") and 6 patients treated with polatuzumab vedotin combined with bendamustine and atolizumab ("Pola-BG") patients. See Figure 1A . The phase II part includes an expanded cohort (20 patients) to evaluate Pola-BG and a randomized cohort (80 patients, 40 patients in each treatment group) that compares Pola-BR with BR alone, according to the response to the last treatment Duration (DOR) stratification (≤12 months compared to >12 months). See Figure 1A .

^* 21天週期^ǂ 週期2-6 表 1B ： Ib/II 期 Pola-BG 治療方案之給藥及投與時程

^* 21天週期^ǂ 週期2-6 表 1C ： II 期 BR 治療方案之給藥及投與時程

^* 21天週期^ǂ 週期2-6All patients received bendamustine ("B") 90 mg/m ² intravenously on days 2 and 3 of cycle 1, and then on days 1 and 2 of subsequent cycles; and Receive rituximab ("R") 375 mg/m ² intravenously on day 1 of cycle or atolizumab intravenously on day 1, 8 and 15 of cycle 1 and day 1 of subsequent cycles Anti ("G") 1000 mg. Those treated with polatuzumab vedotin ("Pola") received 1.8 mg/kg intravenously (IV) on day 2 of cycle 1 and on day 1 of subsequent cycles. Below Table 1A-1C. Patients receive up to 6 21-day cycles of treatment. Table 1A : Dosing and time course of Pola-BR treatment regimen for phase Ib and phase II

^* 21-day cycle ^ǂ cycle 2-6 table 1B : the dosing and time course of the Pola-BG treatment plan for phase Ib/II

^* 21-day cycle ^ǂ cycle 2-6 table 1C : the dosing and administration schedule of phase II BR treatment plan

^* 21 days cycle ^ǂ cycle 2-6

On the day when bendamustine and rituximab were scheduled to be administered, rituximab was administered before bendamustine. On the day when polatuxumab vedotin, bendamustine, and rituximab are scheduled to be administered, rituximab is administered before polatuzumab vedotin, and polatuzumab vedotin is administered before bendamustine. On the day when polatuxumab vedotin, bendamustine, and atolizumab are scheduled to be administered, atolizumab is administered before polatuzumab vedotin, and polatuzumab vedotin is administered before bendamustine. Assessment and endpoint

The primary endpoint of stage Ib is safety and tolerability. The primary endpoint of phase II is at the end of treatment (EOT, 6-8 weeks after day 1 of cycle 6 or the last dose of study treatment) by the Independent Review Committee (IRC) using the modified Lugano response criteria (see Cheson et al. (2014) "Recommendations for initial evaluation, staging, and response assessment of hodgkin and non-hodgkin lymphoma: The Lugano Classification." J. Clin. Oncol. 32:3059-67), with ¹⁸ F-fluorodeoxyglucose-positron emission tomography Scan-The complete response (CR) rate of Pola-BR compared with BR measured by computed tomography (PET-CT). The changes to the Lugano classification are as follows: 1) For patients with bone marrow or an unknown state at baseline, CR assessment based only on imaging methods without confirmation of bone marrow test is classified as partial response (PR); 2) partial response (only By IRC) partial metabolic response of fluorodeoxyglucose-PET and full or partial response of CT are required, otherwise the response according to the modified Lugano criteria (see above) is classified as stable disease.

Secondary endpoints include EOT overall response rate (ORR), best overall response (BOR), duration of response (DOR) and IRC progression-free survival (PFS). Exploratory endpoints include biomarker evaluation of the efficacy of cells of origin (COO) determined by Nanostring Lymphoma Subtype Analysis Test (LST) or Hans Algorithm Standards, and immunohistochemical staining of dual manifestation lymphoma (DEL). User Assessment (INV) DOR and PFS, and OS.

After 3 cycles (mid-term) and at EOT (initial response assessment), the response was evaluated by computed tomography (CT), PET-CT, and bone marrow examination (if necessary to confirm CR). Follow-up CT scans are performed every 6 months for 2 years or until progressive disease (PD) or the patient withdraws.

The National Cancer Institute Adverse Events Commonly Used Terminology Standard (version 4.03) is used to evaluate and score all adverse events (AE) in the entire study. Regardless of the relationship with the study drug, report all AEs from day 1 of cycle 1 to day 90 after the last dose of study drug, including serious adverse events (SAE). After this, continue to report all SAEs indefinitely. Biomarkers

The following methods are used for the exploratory biomarker evaluation of CD79b expression, cell of origin (COO) and dual expression lymphoma (DEL), that is, the dual expression of MYC and BCL2. CD79b

Use AT 107-2 (Serotec) antibody and Ventana Benchmark XT platform to evaluate CD79b tumor cell protein expression by immunohistochemistry (IHC) in a central laboratory. The staining intensity (0-3+) was used to score performance. In addition, by evaluating the continuous measurement of the H-score, the performance range can be evaluated with a larger granularity. The H-score is a weighted scoring system that considers the percentage of tumor cells with 0, 1, 2 or 3+ staining intensity. And the range is 0 to 300. Use the following formula to calculate the H-score of tumor cell staining: H-score = (% at 0 time) × 0 + (% at 1 + time) × 1 + (% at 2 + time) × 2 + (% at 3 + time %)×3. Therefore, this score produces a continuous variable ranging from 0 to 300 (Pfeifer et al. (2015) "Anti-C22 and anti-CD79B antibody drug conjugates are active in different molecular diffuse large B-cell lymphoma subtypes." Leukemia . 29:1578 -86). Cells with H-score staining greater than "0" are considered positive. Cell of Origin (COO)

The samples are sent to Labcorp, where NanoString Reasearch-Only Lymphoma Subtype Test (LST) verification is performed. If COO classification cannot be performed by Nanostring LST ( for example , due to tissue availability), the Hans algorithm using local pathology reports (Hans et al. (2004) "Confirmation of the molecular classification of diffuse large B-cell lymphoma by immunohistochemistry using a tissue microarray." Blood. 103: 275–282) COO was classified by the central pathology evaluation (Histogenex) and IHC. In the analysis, Hans' non-GCB (ie, non-germinal center B cells) was counted as ABC (ie, activated B cells). Dual manifestation lymphoma (DEL)

IHC was performed on the Ventana (Santa Clara, CA) and Ventana Benchmark XT platform using research-only BCL2 (124) mAb and MYC (Y69) IHC. MYC IHC overexpression is defined as ≥40% tumor cell nuclei with positive staining, and BCL2 overexpression is defined as ≥50% tumor cells with cytoplasmic staining intensity ≥2+. Statistical analysis

The phase Ib sample size is determined by a 3+3 design ( see Storer BE. (1989) "Design and analysis of phase I clinical trials." Biometrics. 45:925-37). The sample size of the phase II random cohort was determined based on the assumption that the CR rate difference between Pola-BR and BR was 25%, which allowed zero to be excluded as the lower limit of the 95% confidence interval (CI, 3.8 to 46.2%). For the safety assessment of the phase II part, the sample size of 20 patients in the extended group and 40 patients in each randomized group provides the possibility of ≥85% of observed AEs of ≥1 AE based on the true incidence of 10% and 5%, respectively Sex.

All patients who received ≥1 dose of any study treatment were included in the safety analysis (safety assessment). Efficacy analysis of the intention-to-treat population. result

具有自體幹細胞移植(ASCT)之高劑量化療被計為1療法。DLBCL ， NOS - 彌漫性大B細胞淋巴瘤，未列明；ABC - 活化B細胞樣；GCB - 生髮中心B細胞樣；ECOG PS - 東部腫瘤協作組體力狀態。^* 最後一劑治療後6個月內無響應或進行性疾病113 patients with relapsed-refractory diffuse large B-cell lymphoma (R/R DLBCL) who were not suitable for transplantation were registered. The demographic and disease characteristics of all patients are shown in Table 2 below. For the phase II randomized cohort, patients received an average of two previous treatments, with 1-7 previous treatments in the Pola-BR group and 1-5 previous treatments in the BR group. Table 2 : Baseline characteristics

High-dose chemotherapy with autologous stem cell transplantation (ASCT) is counted as 1 therapy. DLBCL , NOS -diffuse large B-cell lymphoma, not listed; ABC -activated B-cell like; GCB -germinal center B-cell like; ECOG PS -Eastern Cooperative Oncology Group physical status. ^* No response or progressive disease within 6 months after the last dose of treatment

NE =不可估計 * HR(95%CI)=不適用 ǂHR (95% CI) = 0.40 (0.16, 1.01); p= 0.0462 ǂǂHR (95% CI) = 0.44 (0.2, 0.95); p = 0.0321 §HR (95% CI) = 0.36 (0.21, 0.63); p = 0.002 §§HR (95% CI) = 0.34 (0.2, 0.57); p ＜ 0.0001 ** HR (95% CI) = 0.42 (0.24, 0.75); p = 0.0023基於分層分析之 HR 及 p 值。 The safety trial included 6 patients receiving Pola-BR and 6 patients receiving Pola-BG. Twenty-one patients were registered and treated in the Phase II Pola-BG cohort. For the phase II randomized cohort, 40 patients were enrolled in each group, and 39 patients in each group received treatment. See Figure 1B . The baseline characteristics of randomized patients are usually balanced with those who have received an average of 2 previous therapies. Seventy-five percent of Pola-BR and 85% of BR patients were refractory to their last treatment ( for example , did not show a response or progressive disease within 65 months of the last dose of treatment). The response rate and the average effect of EOT end time of the event as shown in Table 3. Table 3 : Summary of efficacy results

NE = not estimable* HR (95%CI) = not applicable ǂHR (95% CI) = 0.40 (0.16, 1.01); p = 0.0462 ǂǂHR (95% CI) = 0.44 (0.2, 0.95); p = 0.0321 §HR (95% CI) = 0.36 (0.21, 0.63); p = 0.002 §§HR (95% CI) = 0.34 (0.2, 0.57); p <0.0001 ** HR (95% CI) = 0.42 (0.24, 0.75) ; p = 0.0023 HR and p value based on stratified analysis .

In the stage Ib Pola-BR group, the CR rate assessed by EOT IRC was 50% (3/6), and all 3 patients remained in remission at an average follow-up of 37.6 months (DOR: 28.9-38.2 months). One non-responder received subsequent treatment and was still alive, and 2 people died of PD. In the combined phase Ib/II Pola-BG group, the CR rate assessed by EOT IRC was 29.6% (8/27). At an average follow-up of 26.9 months, the median PFS (IRC) and OS were 6.3 and 10.8 months, respectively. Two patients continued to consolidate SCT (one autologous and one allogeneic). Without the need for further treatment, 4 patients (15%) recorded a response lasting at least 20 months (range 20.7-22.5 months). At the last follow-up, 8 cases were still alive, 17 cases died (12 cases of PD, 5 cases of AE), and 2 cases were discontinued from the study (1 case determined by the doctor, 1 case of AE).

A preliminary analysis of a randomized cohort of phase II showed that compared with BR, the EOT in the Pola-BR group had a significantly higher CR rate (40.0% vs. 17.5%; P = 0.026; Table 3 ). Compared with the investigator's assessment, a higher proportion of patients are considered not to be evaluated by IRC for EOT response, because patients with evidence of clinical progression but not undergoing follow-up PET-CT cannot be evaluated by IRC. However, this does not affect the evaluation of the CR rate, where the agreement between IRC and the investigator's evaluation is> 90%. Compared with BR, Pola-BR has higher BOR and best CR rate. See Table 3. Without further treatment, 6 patients (15%) had a sustained response duration of ≥20 months.

After an average follow-up of 22.3 months, compared with BR, PFS and OS were significantly improved in Pola-BR, and so was DOR. See Figures 2A and 2B . DOR (IRC: HR 0.40 [95% CI, 0.16 to 1.01]; INV: HR 0.44 [95% CI, 0.20 to 0.95]) and PFS (IRC: HR 0.36 [95% CI, 0.21] assessed by IRC and investigators To 0.63]; INV 0.34 [95% CI, 0.20 to 0.57]) consistent benefits of risk reduction were observed. The IRC assessment of DOR and PFS is slightly longer than the INV assessment, which is mainly caused by the obtaining or not getting the interval in the follow-up confirmation scan required for IRC assessment after the clinical progress determined by INV.

The OS of the Pola-BR group improved significantly, and the risk of death was reduced by 58% (HR, 0.42; 95% CI, 0.24 to 0.75), and the median OS of Pola-BR was longer (12.4 months [95% CI, 9.0 to not assessable [NE ]]), in contrast with BR alone (4.7 months [95% CI, 3.7 to 8.3]). See Figure 2B .

During the follow-up period, 11 patients in the Pola-BR group and 4 patients in the BR group were still alive.

Subgroup analysis after the event showed that all clinical and biological subgroups examined had consistent survival benefits. See Figure 2C , Figure 3A and Figure 3B . Regardless of the patient's refractory status and the number of therapies previously received, the patient's benefit is similar.

In addition, 7 patients (18%) with Pola-BR continued to have a DOR ≥20 months (range 20.0-22.5 months) and maintained complete remission at the last follow-up. One patient received consolidated allogeneic SCT; the other 6 received no additional treatment. Only 2 BR patients (5%) continued to follow up without progress; both received consolidation therapy (one received allogeneic SCT and the other received radiotherapy). safety

In the phase Ib Pola-BR and Ib/II phase Pola-BG groups, treatment delivery and adverse events (AE) were similar to the phase II randomized Pola-BR group. (i) Phase Ib Pola-BR

Among the 6 patients treated in the stage Ib Pola-BR group, the most common AEs in ≥1 patients were loss of appetite, weight loss, diarrhea, hypocalcemia, pneumonia, fever, and thrombocytopenia (all 33.3%) ), hypokalemia and nausea (both 50%) and fatigue (66.7%). One patient had the following grade 3-4 AEs: febrile neutropenia, pneumonia, and thrombocytopenia. No grade 5 AE occurred. (ii) Phase Ib/II Pola-BG

In the combined phase Ib/II Pola-BG group, patients received an average of 4 cycles, and 42.3% of patients completed all treatment cycles. Overall, this is similar to Pola-BR. For all components, the median dose intensity for dose modification and dose delay adjustment is about 99-100%. No patient received a dose reduction of polatuzumab vedotin. The dose of bendamustine was reduced in 26.9% (7/26) of patients. The most common causes of bendamustine dose reduction are neutropenia (15.4%) and fatigue/weakness (7.7%). One patient had a dose reduction due to neutropenia and fatigue (same cycle). Twelve patients (46.2%) had treatment delays. The most common causes of delay in treatment are cytopenias (neutropenia or thrombocytopenia [23.1%]) and infection (15.4%). Two patients had delayed treatment for elevated transaminases, and one patient had delayed treatment for peripheral neuropathy (PN).

The most common AEs that occurred in at least 20% of patients were diarrhea (61.5%), fatigue (53.8%), nausea (53.8%), constipation (42.3%), loss of appetite (42.3%), fever (42.3%), Thrombocytopenia (30.8%), neutropenia (26.9%), anemia (19.2%), vomiting (34.6%) and hypokalemia (23.1%). The most frequently reported grade 3-4 adverse events in at least 10% of patients were neutropenia (26.9%), thrombocytopenia (23.1%), febrile neutropenia (11.5%), and anemia Symptoms (11.5%), nausea (11.5%) and fatigue (11.5%). The incidence of grade 3-4 infection was 23.1%.

Peripheral neuropathy (PN) of all grades occurred in 38.5% of patients, and 15.4% of patients ≥ grade 2. Two patients reported grade 3 muscle weakness, but one of them was consistent with disease progression. Two patients withdrew from all study treatments: one was attributed to grade 2 PN, and the other was attributed to grade 3 muscle weakness.

There are 5 fatal AEs. Three fatal AE infections (pneumonia, fungal pneumonia, and sepsis). The other two are myelodysplastic syndromes (occurring 2 years after the subsequent autotransplantation) and general physical health deterioration. (iii) Fatal AE in Pola-BR and BR

There are 3 fatal AEs in Pola-BR (pneumonia, hemoptysis and pulmonary edema), and 4 in BR (cerebrovascular accident, sepsis [2] and pneumonia) occur within 30 days after treatment.

Fatal AEs that occurred during follow-up (including in the case of PD): Pola-BR (distributed shock [PD], pneumonia [PD], renal failure [PD], intracranial hemorrhage [PD] herpes encephalitis and Sepsis); BR (multiple organ dysfunction [2 cases, all PD], cerebral hemorrhage [PD], leukoencephalopathy [PD], sepsis [PD], heart failure and unexplained death). (iv) Phase II Pola-BR and BR

Among randomized patients, the treatment completion rate of the Pola-BR group was higher than that of the BR (46.2% vs. 23.1%), as was the average completion period (5 vs. 3), mainly due to the higher disease progression rate in the BR group. Progressive disease caused 53.8% of patients treated with BR to stop treatment, and 15.4% of patients treated with Pola-BR stopped treatment. AE was the most common reason for discontinuation of Pola-BR (33.3%; Supplementary Table 1). In the two groups, the most common cause of bendamustine dose reduction was cytopenia (4 Pola-BR, 3 BR). The most common full grade and grade 3-4 AEs are shown in Table 4 below. Although Pola-BR has a higher rate of grade 3-4 anemia and thrombocytopenia, the blood transfusion rates of Pola-BR and BR are similar (red blood cells: 25.6% vs. 20.5%; platelets: 15.4% vs. 15.4%). Pola-BR grade 3-4 neutropenia was higher (46.2% vs. 33.3%), but grade 3-4 infection was similar in the two groups (23.1% Pola-BR and 20.5% BR). The overall incidence of peripheral neuropathy (PN) in Pola-BR patients was 41.0% (16/39) (11 grade 1 and 5 grade 2). At the clinical cut-off, 10 patients resolved and 1 patient improved. PN is the only reason for the decrease in the dose of polatuzumab vedotin. It occurred in 2 (7.7%) patients (both grade 2 PN), and both cases resolved.

*顯示≥20%患者發生的所有級別不良事件，≥10%患者之3-4級AE(安全性評估)。除周圍神經病變外，較佳術語顯示在每個系統器官類別中。 †包括：外周運動神經病，外周感覺神經病，减少之振動感，感覺减退，感覺异常 生物標誌物： CD79b ，起源細胞 (COO) 及雙表現淋巴瘤 (DEL) Fatal AEs occurred in 9 patients who received Pola-BR and 11 patients who received BR. Infection was the most common cause (4 Pola-BR, 5 BR). Table 4. Adverse events in patients treated with Polatuzumab Vedotin combined with bendamustine (Pola-BR) and rituximab compared to bendamustine and rituximab (BR) . ^*

* Shows all grades of adverse events in ≥20% of patients, and grades 3-4 AEs in ≥10% of patients (safety assessment) Except for peripheral neuropathy, better terms are shown in each system organ category. †Including: peripheral motor neuropathy, peripheral sensory neuropathy, reduced vibration sensation, hypoesthesia, paresthesia Biomarkers: CD79b , cell of origin (COO) and dual manifestation lymphoma (DEL)

Of the 83 stained patient samples, 80 (96.4%) had detectable CD79b (immunohistochemistry [IHC] H-score 1-300 or 1 ⁺ -3 ⁺ ). RNA evaluation showed the measurable performance of CD79b in all samples, including 3 samples that were negative by IHC. Refer to Figure 4. In the three samples where CD79b was not detectable by IHC, parallel RNA evaluation showed that the inconsistency with IHC data was significantly higher than the background level of measurable performance. Each point represents a single sample or negative control probe. In the NanostringQCPro Bioconductor R-package, the median level of gene expression level is normalized to the default value. By IHC or gene expression, the response to Pola-based treatment is independent of CD79b levels. As shown in FIG. 5, between the performance of responders and non-responders no significant difference (p = 0.24, Wilcoxon rank sum test with continuity correction on).

ABC - 活化B細胞樣；GCB -生髮中心B細胞樣；n - 數目；Pola-BR - polatuzumab vedotin-BR；BR - 苯達莫司汀加利妥昔單抗；CR - 完全響應；PR - 部分響應；SD -穩定疾病；PD - 進行性疾病；NE - 不可估計 表 6. 按照 COO 亞組的中位 PFS 及 OS 。

PFS - 無進展生存期；OS - 總生存期；COO - 起源細胞；ABC - 活化B細胞樣；GCB -生髮中心B細胞樣；Pola-BR - polatuzumab vedotin-BR；BR - 苯達莫司汀加利妥昔單抗；NE - 不可估計。 *藉由Hans算法分類為非GCB之患者與ABC患者一起分組。 ǂ HR: 0.20 (95% CI, 0.09-0.45) ǂǂ HR: 0.49 (95% CI, 0.23-1.05) § HR: 0.21 (95% CI, 0.09-0.51) §§ HR: 0.57 (95% CI, 0.25-1.31)COO evaluation was performed in 107 patient samples, of which 97 were evaluable. The distribution of COO was 46.4% activated B cells (ABC), 47.4% germinal center B cells (GCB), and 6.2% could not be classified. In the random cohort, the improved results achieved by Pola-BR were observed in both the ABC and GCB subgroups. See Tables 5 and 6. Table 5. Response rate at the end of treatment according to COO subgroup ( investigator's assessment ) .

ABC -activated B cell like; GCB -germinal center B cell like; n -number; Pola-BR -polatuzumab vedotin-BR; BR -bendamustine plus rituximab; CR -complete response; PR -partial Response; SD -stable disease; PD -progressive disease; NE -not estimable Table 6. Median PFS and OS according to COO subgroup .

PFS -progression-free survival; OS -overall survival; COO -cell of origin; ABC -activated B cell-like; GCB -germinal center B-cell like; Pola-BR -polatuzumab vedotin-BR; BR -bendamustine plus Rituximab; NE -not estimable. *Patients classified as non-GCB by Hans algorithm are grouped together with ABC patients. ǂ HR: 0.20 (95% CI, 0.09-0.45) ǂǂ HR: 0.49 (95% CI, 0.23-1.05) § HR: 0.21 (95% CI, 0.09-0.51) §§ HR: 0.57 (95% CI, 0.25 -1.31)

DEL - 雙表現淋巴瘤；Pola-BR - polatuzumab vedotin-BR；BR - 苯達莫司汀加利妥昔單抗；n - 數目；CR - 完全響應；PR - 部分響應；SD - 穩定疾病；PD - 進行性疾病；NE - 不可估計。 表 8. 與 BR 相比，使用 Pola-BR 治療的 DEL 及非 DEL 患者之中位 PFS 及 OS 。

PFS - 無進展生存期；OS - 總生存期Pola-BR - polatuzumab vedotin-BR；BR - 苯達莫司汀加利妥昔單抗；DEL -雙表現淋巴瘤；非 DEL -雙表現淋巴瘤；NE - 不可估計； ǂ HR: 0.35 (95% CI, 0.12-1.11) ǂǂ HR: 0.53 (95% CI, 0.23-1.25) § HR: 0.39 (95% CI, 0.12-1.22) §§ HR: 0.58 (95% CI, 0.24-1.40)In 62 patient samples to assess DEL status, 41.9% were identified as DEL, which is a dual manifestation of MYC and BCL2. In a randomized cohort, improved results achieved by Pola-BR were observed in both DEL and non-DEL patients. See Tables 7 and 8. Table 7. Compared with BR , the response rate of DEL and non- DEL patients treated with Pola-BR at the end of treatment ( investigator's assessment )

DEL -dual manifestation lymphoma; Pola-BR -polatuzumab vedotin-BR; BR -bendamustine plus rituximab; n -number; CR -complete response; PR -partial response; SD -stable disease; PD -Progressive disease; NE -not estimable. Table 8. Median PFS and OS in DEL and non- DEL patients treated with Pola- BR compared with BR .

PFS -progression-free survival; OS -overall survival Pola-BR -polatuzumab vedotin-BR; BR -bendamustine plus rituximab; DEL -dual manifestation lymphoma; non- DEL -dual manifestation lymphoma; NE -not estimable; ǂ HR: 0.35 (95% CI, 0.12-1.11) ǂǂ HR: 0.53 (95% CI, 0.23-1.25) § HR: 0.39 (95% CI, 0.12-1.22) §§ HR: 0.58 ( 95% CI, 0.24-1.40)

R/R DLBCL patients who are not suitable for transplantation, including those patients who have failed autologous SCT, have frustrating results due to limited treatment options. In this randomized comparison, in all COO and DEL subgroups, treatment with Pola-BR resulted in a significant improvement in CR rate, PFS, and OS compared with BR alone. Although 13 patients received additional treatment after progression, patients treated with BR performed poorly, highlighting the limitations of existing drugs. This is the first randomized trial to prove the benefit of OS in R/R DLBCL patients who are not suitable for transplantation.

Compared with BR alone, patients receiving Pola-BR had significantly longer OS (median 12.4 months versus 4.7 months, HR 0.42; 95% CI 0.24, 0.75). All subgroups examined seem to benefit, including refractory patients and patients who have received at least 1, at least 2, at least 3, or more than 3 therapies. In addition, biomarker studies indicate that Pola-BR appears to be beneficial to patients regardless of COO or DEL status. The general performance of CD79b was confirmed, and there was no correlation between the performance level of CD79b and the response. Although the independent contribution of bendamustine to the overall efficacy cannot be measured, the 40% CR rate observed with Pola-BR is significantly higher than the 15% previously reported with the combination of polatuzumab vedotin and anti-CD20 monoclonal antibody (Morschhauser et al (2014) "Preliminary results of a phase II randomized study (ROMULUS) of polatuzumab vedotin (PoV) or pinatuzumab vedotin (PiV) plus rituximab (RTX) in patients (Pts) with relapse / refractory (R / R) non- Hodgkin lymphoma (NHL)." J. Clin. Oncol. 32:15 suppl, 8519). The achievement of CR is related to the improvement of DLBCL, and the higher CR rate observed may partly explain the long-lasting response observed in some patients receiving Pola-BR, many of whom are still disease-free without additional treatment . Pola-BR can be used as an independent treatment or as a bridge for consolidation treatment.

Peripheral neuropathy (PN) is a recognized toxicity associated with monomethyl auristatin E (MMAE)-based antibody-drug conjugates and was closely monitored during this study. Although many patients were exposed to vincristine or platinum in advance, most of the observed PNs are low-grade and reversible, and in relatively few patients, dose reduction or delay is required. Compared with BR, a higher rate of grade 3-4 cytopenia was observed for Pola-BR, but this did not result in a higher risk of infection or the need for blood transfusion.

Obvious and significant benefits of PFS and OS have been observed with Pola-BR, so it is not feasible to conduct a randomized phase III trial. Although this study uses Pola-BR as an independent treatment, considering the observed higher CR rate and prolonged disease control, Pola-BR may provide a valuable treatment option that can be easily provided to a wider range of The patient group.

The combination of Polatuzumab vedotin and BG or BR has tolerable safety characteristics. After an average follow-up of 26.9 months, the CR rate of Pola-BG patients was 29.6%, and the average OS was 10.8 months. 80 patients were randomly assigned (40 in each group) to Pola-BR or BR. After an average follow-up of 22.3 months, the CR rate of Pola-BR patients was significantly increased (40% vs. 17.5%, P = 0.026), PFS and OS were longer (median OS 12.4 vs. 4.7 months, HR 0.42; 95 % CI, 0.24 to 0.75). Compared with BR, patients treated with pola-BR have a higher incidence of grade 3-4 neutropenia, anemia, and thrombocytopenia, but the rate of grade 3-4 infection and blood transfusion is similar. Peripheral neuropathy associated with polatuzumab vedotin is mainly low-grade and resolves in most patients.

Compared with BR treatment, Pola-BR treatment more than doubled overall survival. Treatment with Pola-BR resulted in a 66% reduction in the risk of disease progression or death (measured by the investigator-assessed progression-free survival rate; PFS; HR = 0.34; 95% CI 0.2-0.570; p <0.0001). 40% (16/40) of patients treated with Pola-BR achieved complete remission (CR), while only about 18% (7/40) of patients in the BR group (primary endpoint, by positron emission tomography (PET) ) Measurement; CR rate evaluated by the independent review committee; p = 0.026). In addition, in all subgroups tested, compared with BR, patients treated with Pola-BR achieved a higher CR rate and longer PFS and OS, including those from the cell of origin, germinal center B-cell samples and activated B-cell samples Of patients, these patients are related to the poor prognosis of DLBCL.

About 40% of patients with diffuse large B-cell lymphoma do not respond to the initial treatment or relapse after the initial treatment. This disease trajectory is related to a poor prognosis. Polatuzumab vedotin has shown sustained clinical benefits and may increase the survival rate of this population. The above-mentioned research results show the survival benefit of patients who have relapsed/refractory DLBCL and are not suitable for hematopoietic stem cell transplantation.

It is described in more detail through descriptions and examples for the purpose of clear understanding, but these descriptions and examples should not be regarded as limiting the scope of the present invention. The disclosures of all patents and scientific documents cited in this article are expressly cited as a whole. Table 9 : Amino acid sequence

Figure 1A provides a schematic diagram of the study design of the Phase Ib/II clinical trial described in Example 1 .

Figure 2A(i) provides a Kaplan-Meier chart of the progression-free survival (by INV's PFS) assessed by investigators of patients receiving Pola-BR and patients receiving BR only.

Figure 2A(ii) provides a Kaplan-Meier chart of the progression-free survival evaluated by an independent review committee (through IRC's PFS) for patients receiving Pola-BR and patients receiving BR only.

Figure 2B provides a Kaplan-Meier chart of the overall survival (OS) of patients receiving Pola-BR and those receiving BR only.

Figure 2C provides a forest plot, which shows a subgroup analysis of the overall survival (OS) of patients with various clinical and biological characteristics in the Pola-BR group and the BR group.

Figure 3A provides a forest plot, which shows the subgroup analysis of the researcher-assessed progression-free survival (by INV's PFS) among patients with various clinical and biological characteristics in the Pola-BR group and the BR group .

Figure 3B provides a forest diagram, which shows the subgroup analysis of progression-free survival assessed by the independent review committee (IRC's PFS) of patients with various clinical and biological characteristics in the Pola-BR group and the BR group.

Figure 4 provides the results of the RNA evaluation performed to determine the performance of CD79b in lymph node biopsy samples obtained from patients participating in the clinical trial described in Example 1 .

Figure 5 provides the results of an experiment performed to evaluate the CD79b protein performance level in samples obtained from patients participating in the clinical trial described in Example 1 .

Claims (69)

A method for treating diffuse large B-cell lymphoma (DLBCL) in a person in need, which comprises administering to the person an effective amount of: (a) comprising an immunoconjugate of the following formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence SEQ ID NO: 22; (iii) comprising HVR-H3 of amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 of amino acid sequence of SEQ ID NO: 24; (v) HVR-L2 of amino acid sequence of SEQ ID NO: 25; And (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and in which p is between 1 and 8, (b) an alkylating agent, and (c) an anti-CD20 antibody, wherein the treatment prolongs the Human progression-free survival (PFS). Such as the method of item 1 in the scope of patent application, wherein the treatment prolongs the overall survival (OS) of the person. A method for treating diffuse large B-cell lymphoma (DLBCL) in a person in need, which comprises administering to the person an effective amount of: (a) comprising an immunoconjugate of the following formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence SEQ ID NO: 22; (iii) comprising HVR-H3 of amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 of amino acid sequence of SEQ ID NO: 24; (v) HVR-L2 of amino acid sequence of SEQ ID NO: 25; And (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and in which p is between 1 and 8, (b) an alkylating agent, and (c) an anti-CD20 antibody, wherein the treatment prolongs the Human overall survival (OS). Such as the method of any one of items 1 to 3 in the scope of the patent application, wherein after treatment with the immunoconjugate, the alkylating agent and the anti-CD20 antibody, the person achieves a complete response (CR). The method of any one of items 1 to 4 in the scope of the patent application, wherein the anti-CD79 antibody comprises (i) a VH comprising the amino acid sequence of SEQ ID NO: 19 and (ii) an amine comprising SEQ ID NO: 20 VL of the base acid sequence. Such as the method of any one of the claims 1-5, wherein the anti-CD79 antibody comprises (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 36 and (ii) a heavy chain comprising SEQ ID NO: 35 The light chain of the amino acid sequence. Such as the method according to any one of items 1 to 6 in the scope of patent application, wherein the immunoconjugate is polatuzumab vedotin. Such as the method of any one of items 1-7 in the scope of patent application, wherein the alkylating agent is 4-[5-[bis(2-chloroethyl)amino]-1-methylbenzimidazole-2- Base]butyric acid or its salt. Such as the method of any one of items 1-7 in the scope of patent application, wherein the alkylating agent is bendamustine or its salt. Such as the method of item 9 in the scope of patent application, wherein the alkylating agent is bendamustine-HCl. Such as the method of any one of items 1-10 in the scope of patent application, wherein the anti-CD20 antibody system is rituximab. Such as the method of any one of items 1-11 in the scope of the patent application, wherein the immunoconjugate is administered at a dose of 1.8 mg/kg, the alkylating agent is administered at a dose of 90 mg/m ² and the anti-CD20 The antibody was administered at a dose of 375 mg/m ² . Such as the method of any one of items 1-12 in the scope of the patent application, wherein the immunoconjugate, the alkylating agent and the anti-CD20 antibody are administered for at least 6 21-day cycles, wherein in the 21-day cycle of the first cycle, The immunoconjugate was administered intravenously at a dose of 1.8 mg/kg on the second day, the alkylating agent was administered intravenously at a dose of 90 mg/m ² on the second and third days, and the anti-CD20 antibody The immunoconjugate was administered intravenously at a dose of 375 mg/m ² on day 1, and in every 21-day cycle of cycles 2-6, the immunoconjugate was administered intravenously at a dose of 1.8 mg/kg on day 1. And, the alkylating agent was administered intravenously at a dose of 90 mg/m ² on the first and second days, and the anti-CD20 antibody was administered intravenously at a dose of 375 mg/m ² on the first day. Such as the method of item 13 of the scope of patent application, wherein the immunoconjugate and the alkylating agent are administered sequentially on the second day of the first cycle. Such as the method of claim 14, wherein the immunoconjugate is administered before the alkylating agent. Such as the method of any one of items 13-15 in the scope of patent application, wherein the immunoconjugate, the alkylating agent and the anti-CD20 antibody are administered sequentially on the 1st day of the 2-6 cycle. Such as the method of claim 16, wherein on day 1 of cycles 2-6, the anti-CD20 antibody is administered before the immunoconjugate, and wherein the immunoconjugate is administered before the cyclizing agent. Such as the method of any one of items 13-17 in the scope of patent application, wherein the immunoconjugate, the alkylating agent and the anti-CD20 antibody are further administered after the sixth cycle. Such as the method of item 16 of the scope of patent application, wherein in each cycle after the sixth cycle, the immunoconjugate is administered intravenously at a dose of 1.8 mg/kg on the first day in each 21-day cycle, the The alkylating agent was administered intravenously at a dose of 90 mg/m ² on the first and second days, and the anti-CD20 antibody was administered intravenously at a dose of 375 mg/m ² on the first day. Such as the method of item 18 or 19 of the scope of patent application, wherein in each cycle after the sixth cycle, on the first day of each 21-day cycle, the anti-CD20 antibody is administered before the immunoconjugate, and wherein The immunoconjugate is administered before the alkylating agent. A method for treating diffuse large B-cell lymphoma in a person in need, which comprises administering to the person an effective amount of: (a) comprising an immunoconjugate of the following formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) a heavy chain comprising VH, which VH comprises the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, which VL comprises SEQ ID NO: 20 amino acid sequence, and where p is between 2 and 5, (b) bendamustine or its salt, and (c) rituximab, where the immunoconjugate is at 1.8 mg/kg Dosage administration, the bendamustine or its salt is administered at a dose of 90 mg/m ² , and the rituximab is administered at a dose of 375 mg/m ² , wherein the treatment prolongs the person’s Progressive survival (PFS) and/or overall survival (OS), and wherein: i) the DLBCL is activated B-cell-like DLBCL (ABC-DLBCL) or germinal center B-cell-like DLBCL (GCB-BLBCL); iii) The DLBCL is a dual manifestation lymphoma (DEL) iv) the person has received at least two previous therapies for DLBCL; v) the person has received at least three previous therapies for DLBCL; and/or vi) the person has received More than three previous therapies for DLBCL. Such as the 21st method in the scope of patent application, where p is between 3 and 4. Such as the method of claim 21 or 22, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 36, and wherein the light chain comprises the amino acid sequence of SEQ ID NO: 35. Such as the method of any one of the 21-23 items in the scope of patent application, wherein the bendamustine or its salt is bendamustine-HCl. For example, the method according to any one of items 21-24 in the scope of the patent application, wherein after treatment with the immunoconjugate, the bendamustine or its salt, and the rituximab, the person achieves a complete response ( CR). For example, the method of any one of 21-25 of the scope of patent application, wherein the immunoconjugate, the bendamustine or its salt and the rituximab are administered for at least 6 21-day cycles, wherein In a 21-day cycle of 1 cycle, the immunoconjugate was administered intravenously at a dose of 1.8 mg/kg on the second day, and the bendamustine or its salt was administered at 90 mg/m ² on the second and third days. The dose of rituximab was administered intravenously, and the rituximab was administered intravenously at a dose of 375 mg/m ² on the first day, and in each 21-day cycle after the first cycle, in each 21-day cycle The immunoconjugate was administered intravenously at a dose of 1.8 mg/kg on the first day, and the bendamustine or its salt was administered intravenously at a dose of 90 mg/m ^{2 on} the first and second days And the rituximab was administered intravenously at a dose of 375 mg/m ² on the first day. Such as the method of item 26 in the scope of patent application, wherein the immunoconjugate and the bendamustine or its salt are administered sequentially on the second day of the first cycle. Such as the method of claim 27, wherein the immunoconjugate is administered before the bendamustine or its salt or solvate. Such as the method of any one of items 26-28 in the scope of patent application, wherein the immunoconjugate, the bendamustine or its salt and the rituximab are administered sequentially on the 1st day of the 2-6 cycle versus. Such as the method of claim 29, wherein on day 1 of cycles 2-6, the rituximab is administered before the immunoconjugate, and wherein the immunoconjugate is administered before the alkylating agent versus. For example, the method according to any one of items 26-30 in the scope of patent application, wherein the immunoconjugate, the bendamustine or its salt and the rituximab are further administered after the 6th cycle, and wherein In each cycle after the sixth cycle, in each 21-day cycle, the immunoconjugate was administered intravenously at a dose of 1.8 mg/kg on the first day, and the bendamustine or its salt was administered on the first day and the first day. The rituximab was administered intravenously at a dose of 90 mg/m ^{2 for} 2 days, and the rituximab was administered intravenously at a dose of 375 mg/m ² on the first day. Such as the method of item 31 of the scope of patent application, wherein on the first day of each 21-day cycle of each cycle after the sixth cycle, the rituximab is administered before the immunoconjugate, and wherein the immunoconjugate The substance is administered before the alkylating agent. Such as the method of any one of items 1-32 in the scope of the patent application, wherein the treatment extends PFS to at least about 6 months. Such as the method of any one of items 1-33 in the scope of patent application, wherein the treatment extends PFS to at least 7 months. Such as the method of claim 34, wherein the treatment extends PFS to at least about 7.6 months. Such as the method of item 35 in the scope of patent application, wherein the treatment extends PFS to at least about 8 months. Such as the method of any one of items 1-36 in the scope of the patent application, wherein the treatment extends PFS to at least 11 months. Such as the method of item 37 in the scope of patent application, wherein the treatment extends PFS to at least 11.1 months. Such as the method of any one of claims 2-38 in the scope of patent application, wherein the treatment extends OS to at least about 11 months. Such as the method of any one of claims 2-39 in the scope of patent application, wherein the treatment extends OS to at least about 12 months. Such as the method of claim 40, wherein the treatment prolongs the OS to at least about 12.4 months. Such as the method of any one of items 1-41 in the scope of patent application, wherein the DLBCL is activated B cell-like DLBCL (ABC DLBCL). Such as the method of any one of items 1-41 in the scope of patent application, wherein the DLBCL is a germinal center B cell-like DLBCL (GCB DLBCL). Such as the method of any one of items 1-41 in the scope of patent application, wherein the DLBCL is unlisted (DLBCL-NOS). Such as the method of any one of items 1-41 in the scope of patent application, wherein the DLBCL is a dual manifestation lymphoma (DEL). Such as the method of any one of items 1-45 in the scope of patent application, wherein the DLBCL is relapsed/refractory DLBCL. Such as the method of any one of items 1-46 in the scope of the patent application, wherein the person does not have grade 3b follicular lymphoma, transforming and mild non-Hodgkin lymphoma, or CNS lymphoma. Such as the method of any one of items 1-47 in the scope of patent application, wherein the person has received at least one previous therapy for DLBCL. Such as the method of item 48 in the scope of patent application, wherein the person has received at least two previous therapies for DLBCL. Such as the method of item 49 in the scope of the patent application, in which the person has received at least three previous therapies for DLBCL. Such as the method of item 50 in the scope of patent application, in which the person has received more than three previous therapies for DLBCL. Such as the method of any one of items 1-51 in the scope of patent application, wherein the person is not suitable for autologous stem cell transplantation (ASCT). Such as the method of item 52 in the scope of patent application, where the ASCT is a first-line ASCT, a second-line ASCT, a third-line ASCT, or a third-line ASCT. Such as the method of any one of items 48-53 in the scope of patent application, wherein the person has failed the previous autologous stem cell transplantation. Such as the method of any one of items 48-54 in the scope of patent application, wherein the person has received previous treatment with an anti-CD20 agent. Such as the method of any one of items 48-55 in the scope of patent application, wherein the person has received previous treatment with bendamustine or its salt. Such as the method of any one of items 48-56 in the scope of patent application, wherein the person is refractory to the most recent previous therapy. A kit comprising an immunoconjugate containing the formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iii) ) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25 HVR-L2; (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and p is between 1 and 8, according to items 1-20 and 33-57 of the scope of patent application In any one of the methods, the immunoconjugate is used in combination with an alkylating agent and an anti-CD20 antibody to treat a person with diffuse large B-cell lymphoma (DLBCL) in need. Such as the set of item 58 in the scope of patent application, wherein the anti-CD79b antibody comprises (i) a heavy chain comprising VH, the VH comprising the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, The VL includes the amino acid sequence of SEQ ID NO:20. A kit comprising an immunoconjugate containing the formula

, Wherein Ab is an anti-CD79b antibody, which includes (i) a heavy chain comprising VH, the VH comprising the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, the VL comprising SEQ ID NO: The amino acid sequence of 20, and p is between 2 and 5. According to the method as in any one of the 21-57 patents, the immunoconjugate is combined with bendamustine or its salt and ritux The combination of cilimab is used to treat people with diffuse large B-cell lymphoma (DLBCL) in need. For example, the 60th set of patent application, where p is between 3 and 4. Such as the set of any one of items 58-61 in the scope of patent application, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 36, and wherein the light chain comprises the amino acid sequence of SEQ ID NO: 35. An immunoconjugate comprising the formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 21; (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iii) ) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 23; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25 HVR-L2; (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 26, and p is between 1 and 8. This immunoconjugate is used for the treatment of diffuse large B cells in people in need A method for lymphoma (DLBCL), the method comprising administering an effective amount of the immunoconjugate, alkylating agent, and anti-C20 antibody to the person, wherein the treatment prolongs the person’s progression-free survival (PFS) and/or total Lifetime (OS). For example, the immunoconjugate of the 63rd patent application is used for the method in any one of the 1-20 and 33-57 patents. Such as the immunoconjugate according to the 63rd or 64th patent application, wherein the anti-CD79b antibody comprises (i) a heavy chain comprising VH, the VH comprising the amino acid sequence of SEQ ID NO: 19, and (ii) comprising VL Light chain, the VL includes the amino acid sequence of SEQ ID NO:20. An immunoconjugate comprising the formula

, Wherein Ab is an anti-CD79b antibody, which comprises (i) a heavy chain comprising VH, which VH comprises the amino acid sequence of SEQ ID NO: 19, and (ii) a light chain comprising VL, which VL comprises SEQ ID NO: The amino acid sequence of 20, and p is between 2 and 5. The immunoconjugate is used for the treatment of diffuse large B cell lymphoma (DLBCL) in a person in need, and the method includes administering to the person effective The amount of (a) the immunoconjugate, (b) bendamustine or its salt, and (c) rituximab, wherein the immunoconjugate is administered at a dose of 1.8 mg/kg, the bendam Mustine or its salt was administered at a dose of 90 mg/m ² and the rituximab was administered at a dose of 375 mg/m ² , and wherein the treatment prolonged the person’s progression-free survival (PFS ) And/or overall survival (OS). For example, the immunoconjugate of the 66th patent application is used in the method of any one of the 21-57 patents. Such as the immunoconjugate of item 66 or 67 in the scope of patent application, where p is between 3 and 4. The immunoconjugate according to any one of items 63-68 in the scope of patent application, wherein the anti-CD79 antibody comprises a heavy chain, the heavy chain comprises the amino acid sequence of SEQ ID NO: 36, and the light chain comprises SEQ ID NO : 35 amino acid sequence.

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