Combination treatment of an Anti-cd20 / Anti-cd3 bispecific antibody and chemotherapy in ctdna high risk patients

Abstract

The present invention relates to methods of treating previously untreated diffuse large B-cell lymphoma (DLBCL) defined as high risk according to circulating tumor DNA (ctDNA) by administering galatifamab in combination with chemotherapy.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202480067686.8 (22) Application Date 2024.10.23 (30) Priority Data 63 / 545,997 2023.10.27 US (85) PCT International Application Entering National Phase Date 2026.04.22 (86) PCT International Application Application Data PCT / EP2024 / 079877 2024.10.23 (87) PCT International Application Publication Data WO2025 / 087936 EN 2025.05.01 (71) Applicant: Hofmeister Roche Ltd. Address: Basel, Switzerland Applicant: Genentech Ltd. (72) Inventors: A.T. Barzoos, M.Y. Dolor, K. Humphrey, P.K. Camus, R.A. McDermott, J.M. Ray (74) Patent Agency: Beijing Kunrui Law Firm, 11494 Patent Attorney: Feng Xinqin (51) Int.Cl. A61K 39 / 395 (2006.01) A61K 31 / 282 (2006.01) A61K 31 / 352 (2006.01) A61K 31 / 4745 (2006.01) A61K 31 / 56 (2006.01) A61K 31 / 675 (2006.01) A61K 31 / 7048 (2006.01) A61K 45 / 06(2006.01) A61P 35 / 04(2006.01) C07K 16 / 28(2006.01) (54) Title of Invention Combination therapy of anti-CD20 / anti-CD3 bispecific antibody with chemotherapy in patients at high risk of CTDNA (57) Abstract This invention relates to a method of treating previously untreated diffuse large B-cell lymphoma (DLBCL) defined as high risk based on circulating tumor DNA (ctDNA) by administering glimetuzumab in combination with chemotherapy. Claims (3 pages), Description (71 pages), Sequence Listing (electronic publication), Drawings (4 pages), CN 122070142 A 2026.05.19 CN 1 22 07 01 42 A 1. A method for treating a previously untreated patient with high-risk circulating tumor DNA (ctDNA) diffuse large B-cell lymphoma, the method comprising administering a combination of a bispecific antibody targeting CD3 and CD20, comprising the VH and VL sequences of glimepiride, with chemotherapy, wherein the patient has been identified as high-risk for ctDNA if the ctDNA decreases by less than 2.5 log after one or two cycles of chemotherapy. 2. Claim 1The method, wherein the bispecific antibody targeting CD3 and CD20 comprises the heavy chain of SEQ ID NO: 17, the heavy chain of SEQ ID NO: 18, the two light chains of SEQ ID NO: 20, and the light chain of SEQ ID NO: 19; and wherein the heavy chain and the light chain are assembled to form a first Fab molecule specifically binding to CD20, a second Fab molecule specifically binding to CD3, and a third Fab molecule specifically binding to CD20, and an Fc domain composed of a first subunit and a second subunit capable of stable association. 3. The method according to claim 1 or 2, wherein the chemotherapy is selected from: rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP); rituximab, cyclophosphamide, doxorubicin, and prednisone (R-CHP); or rituximab, ifosfamide, carboplatin, and etoposide phosphate (R-ICE). 4. The method of any one of claims 1 to 3, wherein the bispecific antibody targeting CD3 and CD20 is glimepiride, and the chemotherapy is rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), and wherein the patient is identified as high-risk for ctDNA if the ctDNA decreases by less than 2.5 log after one or two R-CHOP cycles. 5. The method of claim 4, wherein the ctDNA decrease is determined by measuring the amount of ctDNA before or on the day of the first treatment (baseline) and after at least one R-CHOP treatment cycle. 6. The method of claim 4 or 5, wherein the amount of ctDNA is determined on or before day 1 of the first R-CHOP treatment cycle (baseline) and on day 1 of the second R-CHOP treatment cycle. 7. The method of any one of claims 4 to 6, wherein the patient is identified as high-risk for ctDNA if the ctDNA decreases by less than 2 log. 8. A method of treating previously untreated diffuse large B-cell lymphoma, the method comprising: a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least two cycles; b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and c. selecting patients whose ctDNA decreases by <2.5 log between baseline and at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimepiride. 9. The method of claim 8The method of claim 9, wherein step a) comprises administering R-CHOP on day 1 of cycle 1, 2, and optionally cycle 3. 10. The method of claim 9, wherein step a) comprises administering R-CHOP on day 1 of cycles 1 and 2. 11. The method of any one of claims 8 to 10, wherein step b) comprises determining the amount of ctDNA on or before day 1 of cycle 1 (baseline) and day 1 of cycle 2 or day 1 of cycle 3 of the R-CHOP treatment in step a). 12. The method of claim 11, wherein step b) comprises determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2 of the R-CHOP treatment in step a). 13. The method of any one of claims 8 to 12, wherein step c) comprises selecting patients with ctDNA levels <2-log. 14. The method of any one of claims 8 to 13, wherein step c) comprises continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimepiride for a total of eight cycles (cycles 3 to 10). 15. The method of any one of claims 8 to 14, wherein glimepiride is administered at a dose of 2.5 mg on day 8 of cycle 3 and at a dose of 10 mg on day 15, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 30 mg on day 1 of cycles 7 to 10. 16. The method of any one of the preceding claims, wherein the ctDNA is circulating tumor DNA in plasma. 17. The method of any one of the preceding claims, wherein the amount of ctDNA is measured in a blood sample from the patient. 18. The method of claim 17, wherein the amount of ctDNA is measured in genomic DNA isolated from peripheral mononuclear cells or plasma-depleted whole blood (PDWB) or in cell-free DNA (cfDNA) in plasma. 19. The method of claim 18, wherein the ctDNA is measured in cell-free DNA in plasma. 20. The method of any preceding claim, wherein the ctDNA is determined by targeted next-generation sequencing (NGS). 21. The method of any one of claims 4 to 20, wherein rituximab is administered at a dose of 375 mg / m².The treatment is administered intravenously at a dose of 750 mg / m², doxorubicin at a dose of 50 mg / m², vincristine at a dose of 1.4 mg / m² via IV bolus, and prednisone at a dose of 100 mg / day for days 1 to 5. 22. The method of any one of claims 1 to 21, wherein the length of each cycle is 21 days. 23. The method of any one of claims 4 to 22, wherein administration of such treatment to multiple high-risk human patients with ctDNA produces an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%. 24. The method of claim 23, wherein the ORR is at least 95%. 25. The method of any one of claims 1 to 21, wherein administration of such treatment to multiple high-risk human patients with ctDNA produces a complete response (CR) rate of at least 75%, at least 80%, or at least 85%. 26. The method of claim 25, wherein the CR rate is at least 85%. 27. The method of claims 23 and 24 or claims 25 and 26, wherein the ORR or CR rate is determined at the end of treatment. 28. The method of claim 27, wherein the ORR or CR is determined by PET-CT scan. 29. The method of claim 27 or 28, wherein the ORR or CR is determined according to the 2014 Lugano Response Criteria. 30. A bispecific antibody targeting CD3 and CD20 comprising the VH and VL sequences of glimetuzumab, for use in a method of treating previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, the method comprising administering glimetuzumab in combination with chemotherapy to a patient, wherein the patient has been identified as at high risk of ctDNA if the ctDNA decreases by less than 2.5 log after one cycle of chemotherapy. 31. Griffithomab, in a method of treating previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, the method comprising: a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles; b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and c. selecting at baseline and at least one R-CHOP treatment cycle.Patients with a ctDNA reduction of <2.5-log between treatment cycles are continued with R-CHOP and glimetuzumab. 32. Use of a bispecific antibody targeting CD3 and CD20 containing the VH and VL sequences of glimetuzumab in the manufacture of a medicament for treating subjects with previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, wherein the patient is administered glimetuzumab in combination with chemotherapy, wherein the patient has been identified as at high risk of ctDNA if the ctDNA reduction is less than 2.5-log after one chemotherapy cycle. 33. Use of glimetuzumab in the manufacture of a medicament for the treatment of subjects with previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, comprising: a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles; b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one cycle of R-CHOP treatment; and c. selecting patients whose ctDNA decreases by <2.5 log between baseline and at least one cycle of R-CHOP treatment and continuing treatment with R-CHOP and glimetuzumab. 34. The invention as described above. Claims 3 / 3 Page 4 CN 122070142 A Combination therapy of anti-CD20 / anti-CD3 bispecific antibody with chemotherapy in patients at high risk of CTDNA

[0001] This invention relates to a method of treating previously untreated diffuse large B-cell lymphoma (DLBCL) defined as high risk based on circulating tumor DNA (ctDNA) by administering glimepiride in combination with R-CHOP. Background Art

[0002] Diffuse large B-cell lymphoma (DLBCL) is the most common histological subtype of non-Hodgkin lymphoma (NHL), affecting approximately 25,000 new patients annually in the United States and 1.5 million new patients annually worldwide (Rodriguez-Abreu et al. 2007).

[0003] This disease exhibits significant biological and clinical heterogeneity. Although first-line immunochemotherapy using rituximab in combination with cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) is curative in approximately 60% of cases, a subset of patients is refractory to this standard of care (SOC) approach (10%–15%) or relapses after this approach (20%–30%) (Chaganti et al. 2016).

[0004] According to the International Prognostic Index (IPI)By definition, the five-year survival rate for patients with DLBCL ranges from 90% for low-risk patients to less than 40% to 50% for high-risk patients (Mauer et al. 2014; Zhou et al. 2014). Treatment for patients with relapsed / refractory (R / R) DLBCL remains challenging and typically involves treatment with anti-CD20 antibodies and chemotherapy.

[0005] IPIs are widely used to stratify patients with DLBCL based on their age, biochemical parameters, performance status, and disease stage. Existing prognostic scoring systems are not capable of identifying all high-risk patients with sufficient accuracy (International Non-Hodgkin's Lymphoma Prognostic Factors Project 1993; Sehn et al. 2007; Zhou et al. 2014). Furthermore, patients with DLBCL who do not respond well to first-line SOC therapy or relapse after first-line SOC therapy can only be identified after treatment is completed, at which point the potential for ultimate salvage is limited; therefore, it is necessary to optimize effective drugs and rational treatment combinations for all patient subgroups in the front-line setting.

[0006] Up to one-third of patients with 1L LBCL treated with R-CHOP do not achieve long-term response or cure based on disease stage. Predicting outcomes beyond the International Prognostic Index (IPI) score is difficult due to the lack of reproducibility of common biomarkers. ctDNA, as a sensitive biomarker, has the potential to identify newly diagnosed patients who do not respond well to chemoimmunotherapy. Decreased ctDNA levels in patients with 1L diffuse LBCL (DLBCL) after 1–2 therapy cycles are associated with improved survival (Kurtz et al. J Clin Oncol 2018). Whether early intervention can improve outcomes in patients with elevated ctDNA levels during treatment remains unclear. Griffithomab is a CD20xCD3 bispecific antibody that binds to and redirects T cells to eliminate B cells and has been approved by the FDA and EMA as a monotherapy for relapsed / refractory DLBCL after ≥2 lines of prior therapy. Glofit+R-CHOP has a favorable safety and efficacy profile in 1L DLBCL (Topp et al. ASH 2022).

[0007] Glofit+R-CHOP has a manageable safety profile in 1L high-risk LBCL as defined by ctDNA and induces high response rates at interim analysis and EOT. Dynamic treatment-interim risk assessment using ctDNA provides a baseline-independent method for identifying patients with LBCL.Potential for high-risk patients. Specification 1 / 71 pages 5 CN 122070142 A Summary of the Invention

[0008] The present invention is characterized by a method for treating previously untreated patients with high-risk circulating tumor DNA (ctDNA) diffuse large B-cell lymphoma, the method comprising administering a combination of a bispecific antibody targeting CD3 and CD20 containing the VH and VL sequences of glimetuzumab with chemotherapy, wherein the patient has been identified as high-risk for ctDNA if the ctDNA decreases by less than 2.5-log after one or two cycles of chemotherapy.

[0009] In one aspect, the bispecific antibody targeting CD3 and CD20 comprises the heavy chain of SEQ ID NO: 17, the heavy chain of SEQ ID NO: 18, the two light chains of SEQ ID NO: 20, and the light chain of SEQ ID NO: 19; and wherein the heavy and light chains are assembled to form a first Fab molecule that specifically binds to CD20, a second Fab molecule that specifically binds to CD3, and a third Fab molecule that specifically binds to CD20, and an Fc domain consisting of a first subunit and a second subunit capable of stable association.

[0010] In one embodiment, the chemotherapy is selected from: rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP); rituximab, cyclophosphamide, doxorubicin, and prednisone (R-CHP); or rituximab, ifosfamide, carboplatin, and etoposide phosphate (R-ICE).

[0011] In one embodiment, the bispecific antibody targeting CD3 and CD20 is glimetuzumab, and the chemotherapy is rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), and the patient is identified as high-risk for ctDNA if the ctDNA decreases by less than 2.5-log after one or two R-CHOP cycles.

[0012] In one aspect, the invention is characterized by a method of treating a previously untreated patient with high-risk circulating tumor DNA (ctDNA) diffuse large B-cell lymphoma, the method comprising administering glimetuzumab in combination with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), wherein the patient is identified as high-risk for ctDNA if the ctDNA decreases by less than 2.5-log after one or two R-CHOP cycles.

[0013] In one embodiment, ctDNA is determined by measuring the amount of ctDNA before or on the day of the first treatment (baseline) and after at least one R-CHOP treatment cycle.Decrease. In one embodiment, the amount of ctDNA is determined on or before day 1 of the first R-CHOP treatment cycle (baseline) and on day 1 of the second R-CHOP treatment cycle. In another embodiment, the amount of ctDNA is determined on day 1 of the first R-CHOP treatment cycle (baseline) and on day 1 of the second R-CHOP treatment cycle. In one embodiment, if a patient's ctDNA decreases by less than 2 log, the patient is identified as high-risk for ctDNA.

[0014] In another aspect of the invention, a method of treating previously untreated diffuse large B-cell lymphoma is provided, the method comprising:

[0015] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0016] b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0017] c. selecting patients whose ctDNA decreases by <2.5 log between baseline and after at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimepiride.

[0018] In one embodiment, step a) comprises administering R-CHOP on day 1 of cycles 1, 2, and optionally cycle 3. In one embodiment, step a) comprises administering R-CHOP on day 1 of cycles 1 and 2.

[0019] In one embodiment, step b) includes determining the amount of ctDNA on or before day 1 of cycle 1 of R-CHOP treatment in step a) (baseline) and on day 1 of cycle 2 or day 1 of cycle 3. In an example of implementation specification 2 / 71 page 6 CN 122070142 A, step b) includes determining the amount of ctDNA on day 1 of R-CHOP treatment in step a) (baseline) and on day 1 of cycle 2.

[0020] In one embodiment, step c) includes selecting patients with a ctDNA reduction of <2-log. In one embodiment, step c) includes continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6) and adding glimepiride for a total of eight cycles (cycles 3 to 10).

[0021] In one embodiment, glibenclamide is administered at a dose of 2.5 mg on day 8 of cycle 3 and at a dose of 10 mg on day 15, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 10 mg on day 15 of cycles 7 to 10.The drug is administered at a dose of 30 mg per day.

[0022] In any of the above aspects of the invention, ctDNA is circulating tumor DNA in plasma. In one such embodiment, the amount of ctDNA is measured in a blood sample from the patient. In one embodiment, the amount of ctDNA is measured in genomic DNA isolated from peripheral mononuclear cells or plasma-depleted whole blood (PDWB) or in cell-free DNA (cfDNA) in plasma. In one embodiment, ctDNA is measured in cell-free DNA in plasma. In one embodiment, ctDNA is determined by targeted next-generation sequencing (NGS).

[0023] In any of the above aspects of the invention, rituximab is administered intravenously at a dose of 375 mg / m2, cyclophosphamide is administered intravenously at a dose of 750 mg / m2, doxorubicin is administered intravenously at a dose of 50 mg / m2, vincristine is administered via IV bolus at a dose of 1.4 mg / m2, and prednisone is administered at a dose of 100 mg / day from day 1 to day 5.

[0024] In one embodiment of any of the foregoing aspects of the invention, the length of each cycle is 21 days.

[0025] In one embodiment of any of the foregoing aspects of the invention, administration of such treatment to multiple human patients at high risk of ctDNA produces an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%. In one embodiment, the ORR is at least 95%.

[0026] In one embodiment of any of the foregoing aspects of the invention, administration of such treatment to multiple human patients at high risk of ctDNA produces a complete response (CR) rate of at least 75%, at least 80%, or at least 85%. In one embodiment, the CR rate is at least 85%.

[0027] In one embodiment, the ORR or CR rate is determined at the end of treatment. In one embodiment, the ORR or CR rate is determined by PET-CT scan. In one embodiment, the ORR or CR is determined according to the 2014 Lugano response criteria.

[0028] One aspect of the invention relates to a bispecific antibody targeting CD3 and CD20 comprising the VH and VL sequences of glimetuzumab, used in a method of treating previously untreated diffuse large B-cell lymphoma with high (ct)DNA risk, the method comprising administering glimetuzumab in combination with chemotherapy to a patient, wherein the patient has been identified as high-risk for ctDNA if the ctDNA decreases by less than 2.5 log after one cycle of chemotherapy.

[0029] One aspect of the invention relates to glimetuzumab for treatingA method for treating previously untreated diffuse large B-cell lymphoma at high risk of ctDNA includes:

[0030] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0031] b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0032] c. selecting patients whose ctDNA decreases by <2.5 log between baseline and at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimepiride.

[0033] One aspect of the invention relates to the use of a bispecific antibody targeting CD3 and CD20 comprising the VH and VL sequences of glimetuzumab in the manufacture of a medicament for treating a subject with previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, wherein the patient is administered a combination of glimetuzumab with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), wherein the patient has been identified as at high risk of ctDNA if the ctDNA decreases by less than 2.5 log after one R-CHOP cycle.

[0034] One aspect of the invention relates to the use of glimepiride in the manufacture of a medicament for treating a subject with previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, comprising:

[0035] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0036] b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0037] c. selecting patients with a ctDNA reduction of <2.5-log between baseline and at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimepiride.

[0038] Another aspect of the invention relates to the invention as described herein.

[0039] Each embodiment may be incorporated unless the context clearly indicates otherwise. Each embodiment may be applied to various aspects of the invention unless the context clearly indicates otherwise.

[0040] Certain embodiments of the invention will become apparent from the following more detailed description of some preferred embodiments and claims. Brief Description of the Drawings

[0041] Figure 1 is a schematic diagram illustrating the structure of glimetuzumab.

[0042] Figure 2 is a schematic diagram illustrating an overview of the study design as described in Example 1. ctDNA = circulating tumorDNA; R-CHOP = rituximab combined with cyclophosphamide, doxorubicin, vincristine, and prednisone. Patients at high ctDNA risk were those who did not achieve a ≥2-log (100×-) reduction in ctDNA levels between Day 1 of Cycle 1 and Day 1 of Cycle 2. Patients showing a ≥2-log-fold reduction in ctDNA levels during the same time point were considered low-risk ctDNA patients and were screened as unqualified for continued participation for the purposes of this study.

[0043] Figure 3 is a schematic diagram illustrating the dosing regimen as described in Example 1. ctDNA = circulating tumor DNA; R-CHOP = rituximab combined with cyclophosphamide, doxorubicin, vincristine, and prednisone.

[0044] Figure 4 shows the patient response rates (n = 15) in interim analyses performed at the end of Cycle 2 and the end of treatment (EOT). CR, complete response; EOT, end of treatment; ORR, overall response rate; PR, partial response; SD, stable disease. Detailed Description

[0045] The present invention provides a method for treating previously untreated diffuse large B-cell lymphoma (DLBCL) defined as high risk based on circulating tumor DNA (ctDNA) by administration of glimepiride in combination with R-CHOP.

[0046] (i) General Techniques

[0047] Unless otherwise indicated, the practice of the present invention will employ conventional techniques of molecular biology (including recombinant technology), microbiology, cell biology, biochemistry, and immunology, which are within the scope of the art. The following references provide a comprehensive explanation of these techniques: "Molecular Cloning: A Laboratory Manual," 2nd edition (Sambrook et al., 1989); "Oligonucleotide Synthesis" (edited by M. J. Gait, 1984); "Animal Cell Manual 4 / 71 pages 8 CN 122070142 A Culture" (edited by R. I. Freshney, 1987); "Methods in Enzymology" (Academic Press, Inc.); "Current Protocols in Molecular Biology" (edited by FM Ausubel et al., 1987, and regularly updated); "PCR: The Polymerase Chain Reaction" (edited by Mullis et al., 1994); "A Practical Guide to Molecular..."Cloning (Perbal Bernard V., 1988); “Phage Display: A Laboratory Manual” (Barbas et al., 2001).

[0048] (ii) Definitions

[0049] Unless otherwise defined below, the terms used herein are generally as used in the art.

[0050] Unless otherwise specified, the terms “differentiation cluster 20” or “CD20” as used herein refer to any natural CD20 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). CD20 (also known as B-lymphocyte antigen CD20, B-lymphocyte surface antigen B1, Leu-16, Bp35, BM5, and LF5; human protein described in UniProt database registry number P11836) is a hydrophobic transmembrane protein with a molecular weight of about 35 kD and expressed on pre-B and mature B lymphocytes (Valentine, MA et al., J.). Biol. Chem. 264 (1989) 11282-11287; Tedder, TF et al., Proc. Natl. Acad. Sci. U.S.A. 85 (1988) 208-212; Stamenkovic, I. et al., J. Exp. Med. 167 (1988) 1975-1980; Einfeld, DA, et al., EMBO J. 7 (1988) 711-717; Tedder, TF et al., J. Immunol. 142 (1989) 2560-2568). The corresponding human gene is the transmembrane 4 domain, subfamily A member 1, also known as MS4A1. This gene encodes transmembrane 4A. Members of a gene family. Members of this newborn protein family are characterized by common structural features and similar intron / exon splicing boundaries, and exhibit distinct expression patterns in hematopoietic cells and non-lymphoid tissues. This gene encodes a B lymphocyte surface molecule that plays a role in B cell development and differentiation into plasma cells. This family member is located at 11q12 in the cluster of family members. The term covers “full-length”, unprocessed CD20, as well as any form of CD20 produced by intracellular processing. The term also covers naturally occurring variants of CD20, such as splice variants or allelic variants. Alternative splicing of this gene produces two transcript variants encoding the same protein. In one embodiment, CD20 is human CD20.

[0051] Anti-CD20 / Anti-CD3An example of a bispecific antibody is glimetuzumab (WHO Drug Information (International Nonproprietary Name of the drug), Recommended INN: List 83, 2020, Vol. 34, No. 1, p. 39; Proposed INN: List 121 WHO Drug Information, Vol. 33, No. 2, 2019, p. 276, also known as CD20-TCB, RO7082859, or RG6026; CAS #: 2229047-91-8). Gglimetuzumab is a novel T-cell binding bispecific (TCB) full-length antibody with a 2:1 molecular conformation for bivalent binding to CD20 on B cells and monovalent binding to CD3 (particularly the CD3 ε chain (CD3e)) on T cells. Its CD3 binding region is fused to one of the CD20 binding regions in a head-to-tail manner via a flexible linker. This structure endows glimetuzumab with superior in vitro potency compared to other CD20-CD3 bispecific antibodies with a 1:1 configuration, and it exhibits significant antitumor efficacy in a preclinical DLBCL model. The CD20 bivalent nature maintains this potency in the presence of competitive anti-CD20 antibodies, providing opportunities for pre-treatment or co-treatment with these agents. Gglimetuzumab contains an engineered heterodimeric Fc region whose binding to FcgR and C1q is completely eliminated. By simultaneously binding to CD3e of the T cell receptor (TCR) complex on tumor cells expressing human CD20 and T cells, it also induces tumor cell lysis in addition to T cell activation, proliferation, and cytokine release. Gglimetuzumab-mediated B cell lysis is CD20-specific and does not occur in the absence of CD20 expression or in the absence of T cells simultaneously binding to (crosslinking) CD20-expressing cells. In addition to killing, T cells are also activated by CD3 crosslinking, as detected by increased T cell activation markers (CD25 and CD69), cytokine release (IFNγ, TNFα, IL-2, IL-6, IL-10), cytotoxic particle release (granzyme B), and T cell proliferation. A schematic diagram of the molecular structure of glimetuzumab is depicted in Figure 1. The sequences of glimetuzumab are summarized in Table 1.

[0052] Table 1. Sequence IDs of glimetuzumab

[0053]

[0054] As used herein, the term "glimetuzumab" refers to any bispecific anti-CD3 / CD20 having the VH and VL sequences depicted in the table above.Antibodies, including any variants with post-translational modifications. Furthermore, it should be understood that, as used herein, the term “glimetuzumab” also encompasses any biosimilar version approved by a regulatory agency such as, for example, the US FDA or the European EMA. In one embodiment, glimetuzumab is an antibody described in the following literature: WHO Drug Information, proposed INN: List 83, 2020, Vol. 34, No. 1, p. 39; proposed INN: List 121 WHO Drug Information, Vol. 33, No. 2, 2019, p. 276, also known as CD20-TCB, RO7082859, or RG6026; CAS #: 2229047-91-8.

[0055] As used herein, the terms “cytokine release” or “cytokine release” are synonymous with “cytokine storm” or “cytokine release syndrome” (abbreviated as “CRS”) and refer to an increase in the levels of cytokines, particularly tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-2 (IL-2), and / or interleukin-8 (IL-8), in the blood of a subject during or shortly thereafter (e.g., within 1 day) of treatment administration, resulting in adverse symptoms. Cytokine release is defined as a supraphysiological response resulting in or involving the activation of endogenous or infused T cells and / or other immune effector cells following administration of any immunotherapy. Symptoms may be progressive, always include fever at the onset, and may include hypotension, capillary leakage (hypoxia), and peripheral organ dysfunction (Lee et al. 2019). In some cases, such as after administration of CAR-T cells, CRS may also occur several days after administration during CAR-T cell expansion. The incidence and severity usually decrease with subsequent infusions. Symptoms can range from symptomatic discomfort to fatal events and may include fever, chills, dizziness, hypertension, hypotension, dyspnea, restlessness, sweating, flushing, rash, tachycardia, tachypnea, headache, tumor pain, nausea, vomiting, and / or organ failure.

[0056] The term “variable region” or “variable domain” refers to a domain of the antibody heavy or light chain involved in antibody-antigen binding. The variable domains (VH and VL, respectively) of the heavy and light chains of natural antibodies typically have similar structures, with each domain containing four conserved frame 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.

[0057] It is known in the art that glutamine and glutamate at the N-terminus of a recombinant monoclonal antibody can spontaneously cyclize to pyroglutamic acid in vitro. In some embodiments, for each sequence disclosed herein containing N-terminal glutamine and / or glutamate, a corresponding sequence having an N-terminal pyroglutamic acid residue is also considered.

[0058] A “human antibody” is an antibody having an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human or human cell, or derived from the amino acid sequence of a non-human antibody using a complete library of human antibodies or other human antibody-encoding sequences. This definition of a human antibody specifically excludes humanized antibodies containing non-human antigen-binding residues.

[0059] A “humanized” antibody refers to a chimeric antibody containing amino acid residues from non-human HVRs and amino acid residues from human FRs. In some embodiments, the humanized antibody will comprise substantially all of at least one (typically two) variable domains, wherein all or substantially all of the HVRs (e.g., CDRs) correspond to the HVRs of a non-human antibody, and all or substantially all of the FRs correspond to the FRs of a human antibody. The humanized antibody may optionally comprise at least a portion of the antibody constant region derived from a human antibody. An antibody in a “humanized form,” such as a non-human antibody, refers to an antibody that has undergone humanization.

[0060] As used herein, the terms “hypervariant region” or “HVR” refer to each of the regions of antibody variable domains that are hypervariable in the sequence (“complementarity-determining regions” or “CDRs”) and / or form structurally defined loops (“hypervariant loops”) and / or contain antigen contact residues (“antigen contact sites”). Typically, an antibody comprises six HVRs: three in VH (H1, H2, H3) and three in VL (L1, L2, L3). Exemplary HVRs in this article include:

[0061] (a) Highly variable rings appearing at the following 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));

[0062] (b) CDRs appearing at the following amino acid residues: 24–34 (L1), 50–56 (L2), 89–97 (L3), 31–35b (H1), 50–65 (H2), and 95–102 (H3) (Kabat et al., Sequences of Proteins ofImmunological Interest, 5th Edition Public Health Service, National Institutes of Health, Bethesda, MD (1991));

[0063] (c) Antigen contact sites present at amino acid residues 27c-36 (L1), 46-55 (L2), 89-96 (L3), 30-35b (H1), 47-58 (H2) and 93-101 (H3) (MacCallum et al. J. Mol. Biol. 262: 732-745 (1996)); and

[0064] (d) Combinations of (a), (b) and / or (c), including HVR amino acid residues 46-56 (L2), 47-56 (L2), 48-56 (L2), 49-56 (L2), 26-35 (H1), 26-35b (H1), 49-65 (H2), 93-102 (H3) and 94-102 (H3).

[0065] Unless otherwise specified, HVR residues and other residues (e.g., FR residues) in the variable domain are numbered in this document according to Kabat et al., as above.

[0066] “Frame” or “FR” refers to the variable domain residues other than the hypervariable region (HVR) residues. The FR of the variable domain is usually composed of four FR domains: FR1, FR2, FR3 and FR4. Thus, the HVR and FR sequences usually appear in VH (or VL) in the following sequence: FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4. Specification 7 / 71 pages 11 CN 122070142 A

[0067] “Human common frame” is a frame that represents the amino acid residues most frequently present in the selection of human immunoglobulin VL or VH frame sequences. Generally, the selection of human immunoglobulin VL or VH sequences comes from a subgroup of variable domain sequences. Generally, the sequence subgroup is as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Edition, NIH Publication 91-3242, Bethesda MD (1991), Volumes 1-3. In one embodiment, for VL, the subgroup is subgroup κI as described in Kabat et al. (ibid.). In one embodiment, for VH, the subgroup is subgroup III as described in Kabat et al. (ibid.).

[0068] For the purposes of this document, a “recipient human frame” is a frame that contains an amino acid sequence of a light chain variable domain (VL) frame or a heavy chain variable domain (VH) frame derived from a human immunoglobulin frame or a human common frame as defined below. A recipient human frame “derived from” a human immunoglobulin frame or a human common frame may contain the same amino acid sequence as that human immunoglobulin frame or human common frame, or it may contain amino acid sequence variations. In some embodiments, the number of amino acid variations is 10 or fewer, 9 or fewer, 8 or fewer, 7 or fewer, 6 or fewer, 5 or fewer, 4 or fewer, 3 or fewer, or 2 or fewer. In some embodiments, the VL recipient human frame is sequence-identical to the VL human immunoglobulin frame sequence or the human common frame sequence.

[0069] The “class” of an antibody refers to the type of constant domain or constant region possessed by the heavy chain of the antibody. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and some of these antibodies can be further subdivided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to different classes of immunoglobulins are referred to as α, δ, ε, γ, and μ, respectively.

[0070] As used herein, the term IgG “isotype” or “subclass” refers to any subclass of immunoglobulin defined by the chemical and antigenic characteristics of the immunoglobulin constant region.

[0071] The term “Fc domain” or “Fc region” is used herein to define the C-terminal region of the immunoglobulin heavy chain that contains at least a portion of the constant region. This term includes the native sequence Fc region and variant Fc regions. Although the boundaries of the Fc region of the IgG heavy chain may vary slightly, the human IgG heavy chain Fc region is generally defined as extending from Cys226 or Pro230 to the C-terminus of the heavy chain. However, antibodies produced by host cells can undergo post-translational cleavage of one or more (particularly one or two) amino acids from the C-terminus of the heavy chain. Therefore, antibodies produced by host cells by expressing a specific nucleic acid molecule encoding the full-length heavy chain can include the full-length heavy chain, or the antibody can include a cleaved variant of the full-length heavy chain (also referred to herein as a “cleaved variant heavy chain”). This could be the case where the last two C-terminal amino acids of the heavy chain are glycine (G446) and lysine (K447, EU number). Therefore, the C-terminal lysine (Lys447) or C-terminal glycine (Gly446) and lysine (K447) of the Fc region may or may not be present. Unless otherwise stated herein, the amino acid residues in the Fc region or constant region are numbered according to the EU numbering system, also known as the EU index, such as...As described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991 (see above). As used herein, a “subunit” of the Fc domain refers to one of two polypeptides that form a dimer Fc domain, namely a polypeptide that includes the C-terminal constant region of the immunoglobulin heavy chain and is capable of stable self-association. For example, the subunit of the IgG Fc domain contains the constant domains of IgG CH2 and IgG CH3.

[0072] “Modification that promotes association between the first and second subunits of the Fc domain” is the manipulation of the peptide backbone or a post-translational modification of the Fc domain subunit that reduces or prevents the polypeptide containing the Fc domain subunit from associating with the same polypeptide to form a homodimer. As used herein, association-promoting modifications specifically include individual modifications to each of the two Fc domain subunits (i.e., the first and second subunits of the Fc domain) to which association is desired, wherein the modifications are complementary to each other to promote association between the two Fc domain subunits. For example, association-promoting modifications may alter the structure or charge of one or both of the Fc domain subunits to make their association sterically or electrostatically favorable, respectively. Thus, (hetero)dimerization occurs between a polypeptide containing the first Fc domain subunit and a polypeptide containing the second Fc domain subunit, which may differ in the sense of inconsistency in the additional components (e.g., antigen-binding moieties) fused to each subunit. In some embodiments, association-promoting modifications include amino acid mutations, particularly amino acid substitutions, in the Fc domain. In a particular embodiment, association-promoting modifications include individual amino acid mutations, particularly amino acid substitutions, in each of the two subunits of the Fc domain.

[0073] An “activated Fc receptor” is an Fc receptor that, upon binding to the Fc region of an antibody, induces a signaling event that stimulates the receptor-carrying cell to perform an effector function. Activated Fc receptors include FcγRII (CD16a), FcγRI (CD64), FcγRII (CD32), and FcαRI (CD89).

[0074] When referring to antibody use, the term “effector function” refers to those biological activities attributable to the Fc region of the antibody that vary across antibody isotypes. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity (CDC), Fc…Receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cell phagocytosis (ADCP), cytokine secretion, antigen uptake by immune complex-mediated antigen-presenting cells, downregulation of cell surface receptors (e.g., B cell receptors), and B cell activation.

[0075] As used herein, the term “effective cell” refers to a population of lymphocytes that display effector portion receptors (e.g., cytokine receptors) and / or Fc receptors on their surface, which bind to the Fc region of effector portions (e.g., cytokines) and / or antibodies and contribute to the destruction of target cells, such as tumor cells. Effective cells can, for example, mediate cytotoxic or phagocytic responses. Effective cells include, but are not limited to, effector T cells, such as CD8+ cytotoxic T cells, CD4+ helper T cells, γδ T cells, NK cells, lymphokine-activated killer (LAK) cells, and macrophages / monocytes.

[0076] The terms “multispecific” and “bispecific” refer to the ability of an antigen-binding molecule to specifically bind to at least two different antigenic determinants. Typically, a bispecific antigen-binding molecule contains two antigen-binding sites, each of which is specific to a different antigenic determinant. In some embodiments, a bispecific antigen-binding molecule is capable of binding to two antigenic determinants simultaneously, particularly two antigenic determinants expressed on two distinct cell types.

[0077] As used herein, the terms “valence” or “valence state” indicate the presence of a specified number of antigen-binding sites in an antigen-binding molecule. Thus, the term “monovalent binding to antigen” indicates the presence of one (and no more than one) antigen-binding site in an antigen-binding molecule that is specific to the antigen.

[0078] An “antigen-binding site” refers to a site on an antigen-binding molecule that provides interaction with an antigen, namely one or more amino acid residues. For example, the antigen-binding site of an antibody contains amino acid residues from the complementarity determining region (CDR). Natural immunoglobulin molecules typically have two antigen-binding sites, while Fab molecules typically have a single antigen-binding site.

[0079] A “Fab molecule” refers to a protein composed of the VH and CH1 domains of the heavy chain (“Fab heavy chain”) and the VL and CL domains of the light chain (“Fab light chain”).

[0080] “Fusion” means that the components (e.g., the Fab molecule and the Fc domain subunit) are linked directly or via one or more peptide linkers by peptide bonds.

[0081] An “effective amount” of a drug refers to the amount required to produce physiological changes in the cells or tissues to which it is applied.

[0082] The "therapeutic effective amount" of a pharmaceutical agent (e.g., a pharmaceutical composition) refers to the amount that effectively achieves the desired therapeutic or preventive outcome at the necessary dose and time period. A therapeutically effective amount of a pharmaceutical agent, for example, eliminates, reduces, delays, minimizes, or prevents adverse reactions, as described in the instructions for use on page 9 / 71 of CN 122070142 A.

[0083] "Therapeutic agent" means, for example, the active ingredient of a pharmaceutical composition, administered to a subject in an attempt to alter the natural course of a disease in the treated subject, and may be used for prevention or administered during a clinicopathological process. "Immunotherapy agent" refers to a therapeutic agent administered to a subject in an attempt to restore or enhance the subject's immune response (e.g., an immune response against a tumor).

[0084] The term "pharmaceutical composition" refers to a preparation in a form such that the bioactivity of the active ingredient contained therein is effective, and the preparation does not contain any additional components that would have unacceptable toxicity to the subject to whom the composition would be administered.

[0085] "Pharmaceutical carrier" refers to a component of a pharmaceutical composition that is non-toxic to the subject, other than the active ingredient. Pharmaceutically acceptable carriers include, but are not limited to, buffer solutions, excipients, stabilizers, or preservatives.

[0086] The terms “packaging insert” or “instructions for use” are used to refer to instructions typically included in the commercial packaging of a therapeutic product, which include information on indications, usage, dosage, administration, combination therapy, contraindications, and / or warnings for using the therapeutic product.

[0087] The term “combination therapy” as used herein covers combination administration (where two or more therapeutic agents are included in the same or separate formulations); and separate administration, in which case the administration of an antibody as reported herein may occur before, simultaneously with, and / or after the administration of one or more additional therapeutic agents (preferably one or more antibodies).

[0088] A “crossfab” molecule (also referred to as a “Crossfab”) means a Fab molecule in which the variable or constant domains of the Fab heavy chain and light chain are exchanged (i.e., replaced each other), that is, a crossfab molecule comprises a peptide chain consisting of a light chain variable domain VL and a heavy chain constant domain 1CH1 (VL-CH1, in the N-terminal to C-terminal direction), and a peptide chain consisting of a heavy chain variable domain VH and a light chain constant domain CL (VH-CL, in the N-terminal to C-terminal direction). For clarity, in a crossfab molecule in which the variable domains of the Fab light chain and the Fab heavy chain are exchanged, the peptide chain containing the heavy chain constant domain 1CH1 is referred to herein as the “heavy chain” of the (cross)fab molecule. Conversely, in a crossfab molecule in which the constant domains of the Fab light chain and the Fab heavy chain are exchanged, the peptide chain containing the heavy chain variable domain VH is referred to herein as the “heavy chain” of the (cross)fab molecule.The peptide chain is referred to herein as the “heavy chain” of the (cross-linked) Fab molecule.

[0089] In contrast, a “conventional” Fab molecule means a Fab molecule in its natural form, i.e., comprising a heavy chain (VH-CH1, in the N-terminal to C-terminal direction) consisting of variable and constant domains of the heavy chain, and a light chain (VL-CL, in the N-terminal to C-terminal direction) consisting of variable and constant domains of the light chain.

[0090] As used herein, the term “about” refers to a common range of error for the corresponding value that is readily known to those skilled in the art. References to “about” values ​​or parameters herein include (and describe) embodiments relating to that value or parameter itself.

[0091] In the context of this invention, “DLBCL” refers to previously untreated CD20-positive diffuse large B-cell lymphoma (DLBCL), and includes nonspecific (NOS) type DLBCL, including germinal center B-cell-like (GCB) and activated B-cell-like (ABC) / non-GCB types, as well as dual-expressing lymphoma (MYC and BCL2 co-expression), high-grade B-cell lymphoma (HGBCL) with MYC and BCL2 and / or BCL6 translocations, high-grade B-cell lymphoma (HGBCL; e.g., HGBCL NOS, double-hit HGBCL, and triple-hit HGBCL), and new-onset transformed follicular lymphoma.

[0092] “Individual” or “subject” is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cattle, sheep, cats, dogs, and horses), primates (e.g., human and non-human primates, such as monkeys), rabbits, and rodents (e.g., mice and rats). Preferably, the individual or subject is a human. In one case, each subject in the subject population is a human. In another case, each subject in the subject reference population is a human. As used herein, the term “patient” refers to a human individual or subject requiring treatment. Specification 10 / 71 pages 14 CN 122070142 A

[0093] As used herein, “treatment” (and its grammatical variations such as treat or treating) refers to an attempt to alter the natural course of a disease in the treated individual and is a clinical intervention that can be performed for prevention or may be performed during a clinicopathological process. In one embodiment, the term “treatment” refers to treatment with R-CHOP and glimetuzumab. The expected effects of treatment include, but are not limited to, preventing the onset or recurrence of the disease, alleviating symptoms, reducing any direct or indirect pathological consequences of the disease, preventing metastasis, slowing the rate of disease progression, and improving or alleviating symptoms.Disease state, and relief or improvement of prognosis. In some embodiments, the methods of the present invention are used to delay the development of a disease or slow the progression of a disease. In one embodiment, the disease treated is previously untreated DLBCL as defined herein.

[0094] As used herein, “delayed progression” of a condition or disease means delaying, hindering, slowing, stabilizing, and / or postponing the development of a disease or condition (e.g., DLBCL). Such delay can have different durations depending on medical history and / or the individual being treated. It will be apparent to those skilled in the art that sufficient or significant delay can actually cover prevention, as the individual will not develop the disease. For example, in advanced cancer, the development of central nervous system (CNS) metastases may be delayed.

[0095] As used herein, “administration” means a method of giving a subject a dose of a compound (e.g., glimepiride) or composition (e.g., a pharmaceutical composition, such as a pharmaceutical composition comprising glimepiride). The compounds and / or compositions utilized in the methods described herein may be administered intravenously (e.g., by intravenous infusion).

[0096] As used herein, “target dose” refers to the dose of glimetuzumab to achieve the therapeutic effect, i.e., to achieve the desired clinical efficacy. Possible target doses for glimetuzumab have been found to be 16 mg or 30 mg.

[0097] “Using the target dose consistently or as intended” and “treatment regimen without using an escalating dosing regimen” refer to a dosing regimen that uses the same dose in the first and second cycles and optionally any subsequent treatment cycles, as opposed to escalating dosing, which uses a lower dose in the first few treatment cycles and only reaches the target dose in the second or subsequent treatment cycles.

[0098] As used herein, the term “treatment cycle” or “cycle” (abbreviated as “C”) means a process of repeating one or more doses of a therapeutic agent (e.g., R-CHOP and / or glimetuzumab) at a regular schedule, optionally with rest periods (no treatment). In one aspect of the invention, a third treatment cycle comprises a first and second dose of glimetuzumab, followed by a rest period. In one such embodiment, the third treatment cycle includes a first dose of glimepiride on day 8 of the third cycle and a second dose of glimepiride on day 15 of the third cycle, followed by a 6-day rest period. In one embodiment, the subsequent two cycles (cycles 4 to 6) include a dose of glimepiride administered on day 8 of that cycle, followed by a 13-day rest period. In one embodiment, a treatment cycle comprises 21 days. In another embodiment, a treatment cycle comprises 14 days. Treatment regimens according to the invention may include 6 or more treatment cycles, particularly 10 treatment cycles. In some embodiments, a treatment cycle is referred to as a “dosing cycle”.

[0099] “Individual response” or “response” may be assessed using any endpoint that indicates benefit to the subject, including but not limited to (1) inhibiting disease progression (e.g., DLBCL progression) to some extent, including slowing and completely blocking it; (2) reducing tumor size; (3) inhibiting (i.e., reducing, slowing or completely stopping) cancer cell infiltration into adjacent peripheral organs and / or tissues; (4) inhibiting (i.e., reducing, slowing or completely stopping) metastasis; (5) alleviating one or more symptoms associated with DLBCL to some extent; (6) increasing or prolonging survival, including overall survival and progression-free survival; and / or (7) reducing mortality at a given time point after treatment.

[0100] As used herein, “end of treatment” or “EOT” is defined as the time when a patient has completed all study treatment cycles, i.e., 2 cycles of R-CHOP, 4 cycles of R-CHOP and glimetuzumab, and 4 cycles of glimetuzumab treatment (i.e., all 10 cycles depicted in Figures 2 and 3). Instructions for Use, 11 / 71 pages, 15 CN 122070142 A

[0101] As used herein, “complete remission” or “CR” means the disappearance of all target lesions. In one embodiment, the NHL remission criteria are used to determine CR. (Lugano Classification, Cheson et al. J Clin Oncol. 20 Sep 2014; 32(27): 3059–3067.). In one embodiment, the CR rate is defined as the proportion of participants who achieve CR at the end of treatment (i.e., within the 10 cycles depicted in Figures 2 and 3), determined by the investigator according to the Lugano criteria. The CR rate is defined as the percentage of patients with CR at EOT.

[0102] In one embodiment, CR is defined as complete metabolic remission, as determined by PET / CT of lymph nodes and extralymphatic sites, wherein the score is 1, 2, or 3, with or without residual mass at 5PS, wherein PET 5PS: 1 = no uptake above background; 2 = uptake > mediastinum; 3 = uptake > mediastinum, but ≤ liver; 4 = adequate uptake > liver; 5 = significantly higher uptake than liver and / or new lesions; X = new uptake area unlikely to be associated with lymphoma. In one embodiment, CR is defined as complete radiological remission determined by CT of lymph nodes and extralymphatic sites, wherein the target nodule / nodular mass must shrink to ≤ 1.5 cm in LDi (longest transverse diameter of the lesion) and no extralymphatic disease site remains.

[0103] As used herein, “partial response” or “PR” refers to partial metabolic response and, according to Luga no.Classification, Cheson et al. J Clin Oncol. 20 Sep 2014; 32(27): 3059–3067. In one embodiment, PR is determined by PET / CT of lymph nodes and extranodal sites, and / or partial remission is determined by CT of lymph nodes and extranodal sites. In one embodiment, partial metabolic remission is defined by a score of 4 or 5b, a reduction in uptake compared to baseline, and residual mass of any size is determined by PET / CT of lymph nodes and extranodal sites, where PET 5PS: 1 = no uptake above background; 2 = uptake > mediastinum; 3 = uptake > mediastinum, but ≤ liver; 4 = adequate uptake > liver; 5 = significantly higher uptake than liver and / or new lesions; X = new uptake area unlikely to be associated with lymphoma. In one embodiment, partial remission is defined as a reduction of at least 50% in the product of diameters (SPD) of up to 6 measurable nodules and extranodal sites, with baseline SPD as a reference.

[0104] As used herein, “objective response rate (ORR)” is defined as the sum of the partial response (PR) rate and the complete response (CR) rate at the end of treatment (EOT). In one embodiment, ORR is assessed based on the Lugano classification (Cheson et al. J Clin Oncol. 20 Sep 2014; 32(27): 3059-3067). In one embodiment, ORR is measured by PET-CT scan. In one embodiment, the analysis of ORR is the same as that described for the primary efficacy endpoint of CR rate at EOT measured by PET / CT scan.

[0105] “Progression-free survival” (PFS) is defined as the time from the first treatment with glimetuzumab to the first occurrence, relapse, or death from any cause of disease progression (whichever occurs first). In one embodiment, PFS is assessed based on the Lugano classification (Cheson et al. J Clin Oncol. 20 Sep 2014; 32(27): 3059–3067.). In one embodiment, the distribution of PFS, median (if analytically feasible) 6-month and 1-year PFS will be estimated using the Kaplan-Meier method (Kaplan and Meier 1958), where the 95% CI of the median PFS is constructed using the Brookmeyer-Crowley method (Brookmeyer and Crowley 1982).

[0106] "Total Survival" (OS)It is defined as the time from the first use of glimetuzumab treatment to death from any cause.

[0107] "Duration of objective response (DOR)" is defined as the time from the first occurrence of a documented objective response (CR or PR) to disease progression, relapse, or death from any cause (whichever occurs first), as determined by the investigator according to the Lugano criteria (Cheson et al. J Clin Oncol. 20 Sep 20, 2014; 32(27): 3059–3067.). In one embodiment, DOR is measured by PET / CT scan.

[0108] As used herein, "stable disease" or "SD" refers to a minimum SLD since the start of treatment, where the target lesion has neither sufficiently shrunk to meet PR nor sufficiently increased to meet PD.

[0109] As used herein, “disease progression” or “PD” means an increase of at least 20% in the target lesion’s SLD relative to the minimum SLD recorded since the start of treatment, or an increase of at least 50% in the target lesion’s SPD relative to the minimum SPD recorded since the start of treatment, or the appearance of one or more new lesions.

[0110] As used herein, “infusion-related reaction,” “IRR,” or infusion-related adverse event is an adverse event that occurs in a patient or subject during or within 24 hours after administration of a drug (e.g., glimepiride or rituximab). IRR can be graded from 1 to 5 according to, for example, NCI CTCAE v.4.

[0111] As used herein, the term “R-CHOP” means rituximab plus ifosfamide, carboplatin, etoposide, or etoposide phosphate.

[0112] As used herein, the term “ctDNA” means circulating tumor DNA in the bloodstream that is not cellular. It refers to fragments of DNA from cancer cells and tumors, and can be found in plasma or serum. In one embodiment, ctDNA is plasma ctDNA.

[0113] As used herein, “high-risk ctDNA” refers to a patient with a high amount of residual circulating tumor DNA after initial first-line treatment with standard of care (R-CHOP) for DLBCL. In one embodiment, a patient is identified as high-risk ctDNA if the patient’s ctDNA decrease is less than 2.5 log after at least one R-CHOP treatment cycle at or before treatment initiation (i.e., at or before cycle 1, baseline). In one embodiment, a patient is identified as high-risk ctDNA if the patient’s ctDNA decrease is less than 2 log after at least one R-CHOP treatment cycle at or before treatment initiation (i.e., at or before cycle 1, baseline).High risk. In the context of this invention, “at or before cycle 1” means that the baseline ctDNA level is preferably determined on day 1 of cycle 1 of R-CHOP treatment or shortly before day 1 of cycle 1 of R-CHOP treatment (e.g., up to 1, 2, 3, 4, or 5 days prior, preferably up to 1 or 2 days prior).

[0114] In one embodiment, a high-risk ctDNA patient is one who fails to achieve a ≥ 2 log-fold reduction in ctDNA levels between day 1 of cycle 1 of R-CHOP treatment and day 1 of cycle 2.

[0115] As used herein, “2.5 log reduction” and “2 log reduction” refer to a reduction in the amount of ctDNA measured in the patient’s plasma before the start of treatment and after at least one cycle of R-CHOP treatment. For example, a “2 log reduction” in ctDNA means a 100-fold reduction in plasma ctDNA levels.

[0116] As used herein, the “2014 Lugano response criteria” refers to the 2014 Lugano response criteria for malignant lymphomas described in Cheson BD, Fisher RI, Barrington SF, et al., Recommendations for initial evaluation, staging and response assessment of Hodgkin and non-Hodgkin lymphoma: the Lugano classification. J Clin Oncol 2014;32:1-9.

[0117] (iii) Combination therapy of anti-CD20 / anti-CD3 bispecific antibody with anti-CD20 antibody and chemotherapy in patients at high risk of ctDNA

[0118] The present invention is characterized by a method of treating a previously untreated patient with high risk of circulating tumor DNA (ctDNA) of diffuse large B-cell lymphoma, the method comprising administering a combination of a bispecific antibody targeting CD3 and CD20 comprising the VH and VL sequences of glimetuzumab with chemotherapy, wherein the patient has been identified as at high risk of ctDNA if the ctDNA decreases by less than 2.5 log after one or two cycles of chemotherapy.

[0119] In one aspect, the bispecific antibody targeting CD3 and CD20 comprises the heavy chain of SEQ ID NO: 17, the heavy chain of SEQ ID NO: 18, the two light chains of SEQ ID NO: 20 and the light chain of SEQ ID NO: 19; and whereinThe heavy and light chains are assembled to form a first Fab molecule that specifically binds to CD20, a second Fab molecule that specifically binds to CD3, and a third Fab molecule that specifically binds to CD20, and an Fc domain consisting of a first subunit capable of stable association and a second subunit.

[0120] In one embodiment, the chemotherapy is selected from: rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP); rituximab, cyclophosphamide, doxorubicin, and prednisone (R-CHP); or rituximab, ifosfamide, carboplatin, and etoposide phosphate (R-ICE).

[0121] In one embodiment, ctDNA reduction is determined by measuring the amount of ctDNA before or on the day of the first treatment (baseline) and after at least one chemotherapy cycle. In one embodiment, the amount of ctDNA is determined on or before day 1 of the first chemotherapy cycle (baseline) and on day 1 of the second chemotherapy cycle. In one embodiment, the amount of ctDNA is determined on day 1 of the first chemotherapy cycle (baseline) and on day 1 of the second chemotherapy cycle. In one embodiment, a patient is identified as high-risk for ctDNA if the patient's ctDNA decreases by less than 2 log.

[0122] In one embodiment, the bispecific antibody targeting CD3 and CD20 is glimetuzumab.

[0123] In another aspect of the invention, a method of treating previously untreated diffuse large B-cell lymphoma is provided, the method comprising:

[0124] a. administering chemotherapy for at least 2 cycles;

[0125] b. determining the amount of ctDNA on or before cycle 1 (baseline) and after at least one chemotherapy cycle; and

[0126] c. selecting patients whose ctDNA decreases by <2.5 log between baseline and after at least one chemotherapy cycle and continuing treatment with chemotherapy and glimetuzumab.

[0127] In one embodiment, step a) includes administering chemotherapy on day 1 of cycle 1, 2, and optionally cycle 3. In one embodiment, step a) includes administering chemotherapy on day 1 of cycles 1 and 2.

[0128] In one embodiment, step b) includes determining the amount of ctDNA on or before day 1 of cycle 1 (baseline) of chemotherapy treatment in step a) and on day 1 of cycle 2 or day 1 of cycle 3. In one embodiment, step b) includes determining the amount of ctDNA on day 1 of chemotherapy treatment in step a).The amount of ctDNA is determined on day 1 (baseline) and day 1 of cycle 2.

[0129] In one embodiment, step c) includes selecting patients with a ctDNA reduction of <2-log. In one embodiment, step c) includes continuing chemotherapy for a total of six cycles (cycles 1 to 6) and adding glimetuzumab for a total of eight cycles (cycles 3 to 10).

[0130] In one embodiment, glimetuzumab is administered at a dose of 2.5 mg on day 8 of cycle 3 and at a dose of 10 mg on day 15, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 30 mg on day 1 of cycles 7 to 10.

[0131] In any of the foregoing aspects of the invention, ctDNA is circulating tumor DNA in plasma. In one such embodiment, the amount of ctDNA is measured in a blood sample from the patient. In one embodiment, the amount of ctDNA is measured in genomic DNA isolated from peripheral mononuclear cells or plasma-depleted whole blood (PDWB) or in cell-free DNA (cfDNA) in plasma. In one embodiment, ctDNA is measured in cell-free DNA in plasma. In one embodiment, ctDNA is determined by targeted next-generation sequencing (NGS).

[0132] In any of the above aspects of the invention, the length of each cycle is 21 days.

[0133] In one embodiment, the bispecific antibody targeting CD3 and CD20 is glimepiride, and the chemotherapy is rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), and wherein if the patient's ctDNA decreases by less than 2.5 log after one or two R-CHOP cycles, the patient has been identified as having a high risk of ctDNA.

[0134] In one aspect, the invention is characterized by a method for treating a previously untreated patient with diffuse large B-cell lymphoma at high risk of circulating tumor DNA (ctDNA), the method comprising administering glimepiride in combination with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), wherein the patient is identified as high-risk for ctDNA if the ctDNA decreases by less than 2.5 log after one or two R-CHOP cycles. ctDNA is a novel biomarker that offers the possibility of early and sensitive identification of patients with DLBCL at a high risk of first-line treatment failure. In the first-line setting, patients withA subset of patients with DLBCL had poor prognoses after treatment with R-CHOP immunochemotherapy. The inventors of this invention discovered that treatment of previously untreated patients with diffuse large B-cell lymphoma at high ctDNA risk using a combination of glimetuzumab with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) produced unexpectedly high overall response and complete response rates.

[0135] In one embodiment, ctDNA reduction is determined by measuring the amount of ctDNA before or on the day of the first treatment (baseline) and after at least one R-CHOP treatment cycle. In one embodiment, the amount of ctDNA is determined on or before day 1 of the first R-CHOP treatment cycle (baseline) and day 1 of the second R-CHOP treatment cycle. In one embodiment, the amount of ctDNA is determined on day 1 of the first R-CHOP treatment cycle (baseline) and day 1 of the second R-CHOP treatment cycle. In one embodiment, a patient is identified as high ctDNA risk if the patient's ctDNA reduction is less than 2 log. In one embodiment, if the ctDNA level decreases by less than 2 log-fold between day 1 of cycle 1 and day 1 of cycle 2 as determined by an AOA-NHL test, the patient is identified as high-risk for ctDNA.

[0136] In another aspect of the invention, a method of treating previously untreated diffuse large B-cell lymphoma is provided, the method comprising:

[0137] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0138] b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0139] c. selecting patients with a ctDNA decrease of <2.5 log between baseline and after at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimepiride.

[0140] In another aspect of the invention, a method for treating previously untreated diffuse large B-cell lymphoma is provided, the method comprising:

[0141] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0142] b. determining the amount of ctDNA after cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0143] c. selecting patients whose ctDNA decreases by <2.5 log between baseline and at least one R-CHOP treatment cycle, and continuing R-CHOP treatment.Treatment with glimepiride.

[0144] In one embodiment, step a) includes administering R-CHOP on day 1 of cycle 1, 2, and optionally cycle 3. In one embodiment, step a) includes administering R-CHOP on day 1 of cycles 1 and 2.

[0145] In one embodiment, step b) includes determining the amount of ctDNA on day 1 or before (baseline) of cycle 1 of R-CHOP treatment in step a) and on day 1 of cycle 2 or cycle 3. In one embodiment, step b) includes determining the amount of ctDNA on day 1 (baseline) of cycle 1 of R-CHOP treatment in step a) and on day 1 of cycle 2. Specification 15 / 71 pages 19 CN 122070142 A

[0146] In one embodiment, step c) includes selecting patients with a ctDNA reduction of <2-log. In one embodiment, step c) includes continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6) and adding glimepiride for a total of eight cycles (cycles 3 to 10).

[0147] In one aspect of the invention, a third treatment cycle includes a first dose and a second dose of glimepiride, followed by a rest period. In one such embodiment, the third treatment cycle includes a first dose of glimepiride on day 8 of the third cycle and a second dose of glimepiride on day 15 of the third cycle, followed by a 6-day rest. In one embodiment, the subsequent two cycles (cycles 4 to 6) include a dose of glimepiride administered on day 8 of the cycle, followed by a 13-day rest. In one embodiment, the remaining subsequent cycles (cycles 7 to 10) include a dose of glimepiride administered on day 1 of the cycle, followed by a 20-day rest.

[0148] In one embodiment, a treatment cycle comprises 21 days. In another embodiment, a treatment cycle comprises 14 days. Treatment regimens according to the invention may include six or more treatment cycles, particularly ten treatment cycles.

[0149] In one embodiment, glimepiride is administered at a dose of 2.5 mg on day 8 of cycle 3, at a dose of 10 mg on day 15 of cycle 3, at a dose of 30 mg on day 8 of cycles 4 through 6, and at a dose of 30 mg on day 1 of cycles 7 through 10.

[0150] In another embodiment, glimepiride is administered at a dose of 0.5 mg on day 4 of cycle 3 and at a dose of 2.5 mg on day 8 of cycle 3.The dosage was as follows: 10 mg on day 15 of cycle 3, 30 mg on day 8 of cycles 4 through 6, and 30 mg on day 1 of cycles 7 through 10.

[0151] In another embodiment, glimetuzumab was administered at a dose of 0.5 mg on day 8 of cycle 3, 2 mg on day 9, 10 mg on day 15 of cycle 3, 30 mg on day 8 of cycles 4 through 6, and 30 mg on day 1 of cycles 7 through 10.

[0152] One aspect of the present invention relates to a bispecific antibody targeting CD3 and CD20 comprising the VH and VL sequences of glimetuzumab, in a method of treating previously untreated diffuse large B-cell lymphoma at high (ct)DNA risk, the method comprising administering glimetuzumab in combination with chemotherapy to a patient, wherein the patient has been identified as at high ctDNA risk if the ctDNA decreases by less than 2.5 log after one cycle of chemotherapy.

[0153] In one aspect, the bispecific antibody targeting CD3 and CD20 comprises the heavy chain of SEQ ID NO: 17, the heavy chain of SEQ ID NO: 18, the two light chains of SEQ ID NO: 20, and the light chain of SEQ ID NO: 19; and wherein the heavy chain and light chain are assembled to form a first Fab molecule that specifically binds to CD20, a second Fab molecule that specifically binds to CD3, and a third Fab molecule that specifically binds to CD20, and an Fc domain consisting of a first subunit and a second subunit capable of stable association.

[0154] In one embodiment, the chemotherapy is selected from: rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP); rituximab, cyclophosphamide, doxorubicin, and prednisone (R-CHP); or rituximab, ifosfamide, carboplatin, and etoposide phosphate (R-ICE).

[0155] In one embodiment, glimetuzumab is provided in a method of treating previously untreated diffuse large B-cell lymphoma with high (ct)DNA risk, the method comprising administering glimetuzumab in combination with chemotherapy to a patient, wherein the patient has been identified as high-risk for ctDNA if the ctDNA decreases by less than 2.5 log after one cycle of chemotherapy.

[0156] In one embodiment, the patient's ctDNA decreases by less than 2 log. Specification 16 / 71 pages 20 CN122070142 A

[0157] In one embodiment, the bispecific antibody targeting CD3 and CD20 is glimetuzumab, and the chemotherapy is rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), and wherein the patient has been identified as high-risk for ctDNA if the ctDNA decreases by less than 2.5-log after one or two R-CHOP cycles.

[0158] One aspect of the invention relates to glimetuzumab in a method of treating previously untreated diffuse large B-cell lymphoma with high (ct)DNA risk, the method comprising administering glimetuzumab to a patient in combination with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), wherein the patient has been identified as high-risk for ctDNA if the ctDNA decreases by less than 2.5-log after one R-CHOP cycle.

[0159] In one embodiment, ctDNA reduction is determined by measuring the amount of ctDNA before or on the day of the first treatment (baseline) and after at least one R-CHOP treatment cycle. In one embodiment, the amount of ctDNA is determined on or before day 1 of the first R-CHOP treatment cycle (baseline) and day 1 of the second R-CHOP treatment cycle. In another embodiment, the amount of ctDNA is determined on day 1 of the first R-CHOP treatment cycle (baseline) and day 1 of the second R-CHOP treatment cycle. In one embodiment, a patient is identified as high-risk for ctDNA if the reduction in ctDNA is less than 2 log. In one embodiment, a patient is identified as high-risk for ctDNA if the reduction in ctDNA level between day 1 of cycle 1 and day 1 of cycle 2, as determined by the AOA-NHL test, is less than 2 log-fold.

[0160] One aspect of the invention relates to glimetuzumab for the treatment of previously untreated diffuse large B-cell lymphoma at high (ct)DNA risk, the method comprising administering glimetuzumab to a patient in combination with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), wherein the patient is identified as high-risk for ctDNA if the patient's ctDNA reduction is less than 2-log, wherein the ctDNA reduction is measured on or before day 1 of the first R-CHOP treatment cycle (baseline) and on day 1 of the second R-CHOP treatment cycle. In one such embodiment, the ctDNA reduction is measured on day 1 of the first R-CHOP treatment cycle (baseline) and on day 1 of the second R-CHOP treatment cycle.Measurements taken on day 1 of a R-CHOP treatment cycle.

[0161] One aspect of the invention relates to glimetuzumab for the treatment of previously untreated diffuse large B-cell lymphoma, the method comprising:

[0162] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0163] b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0164] c. selecting patients whose ctDNA decreases by <2.5 log between baseline and after at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimetuzumab.

[0165] One aspect of the invention relates to glimetuzumab, in a method of treating previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, the method comprising:

[0166] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0167] b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0168] c. selecting patients whose ctDNA reduction is <2.5 log between baseline and at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimetuzumab. Specification 17 / 71 pages 21 CN 122070142 A

[0169] In another aspect of the invention, glimetuzumab is provided as a method of treating previously untreated diffuse large B-cell lymphoma, the method comprising:

[0170] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0171] b. determining the amount of ctDNA after cycle 1 (baseline) and at least one R-CHOP treatment cycle; and

[0172] c. selecting patients whose ctDNA decreases by <2.5-log between baseline and at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimetuzumab.

[0173] In one embodiment, step a) comprises administering R-CHOP on day 1 of cycles 1, 2, and optionally cycle 3. In one embodiment, step a) comprises administering R-CHOP on day 1 of cycles 1 and 2.

[0174] In one embodiment, step b) includes day 1 or before (baseline) of cycle 1 of the R-CHOP treatment in step a) and day 2 of cycle 2.The amount of ctDNA is determined on day 1 or day 1 of cycle 3. In one embodiment, step b) includes determining the amount of ctDNA on day 1 (baseline) of cycle 1 and day 1 of cycle 2 of R-CHOP treatment in step a).

[0175] In one embodiment, step c) includes selecting patients with a ctDNA reduction of <2-log. In one embodiment, step c) includes continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6) and adding glimepiride for a total of eight cycles (cycles 3 to 10).

[0176] In one aspect of the invention, a method of using glimetuzumab for treating previously untreated diffuse large B-cell lymphoma is provided, the method comprising:

[0177] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least two cycles;

[0178] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0179] c. selecting patients whose ctDNA decreases by <2-log between baseline and one cycle of R-CHOP treatment and continuing treatment with R-CHOP and glimetuzumab.

[0180] In another aspect of the invention, a method of using glimetuzumab for treating previously untreated diffuse large B-cell lymphoma is provided, the method comprising:

[0181] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0182] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0183] c. selecting patients whose ctDNA decreases <2-log between baseline and one R-CHOP treatment cycle, and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of 8 cycles (cycles 3 to 10).

[0184] In another aspect of the invention, a method of using rituximab for treating previously untreated diffuse large B-cell lymphoma is provided, the method comprising:

[0185] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for two cycles;

[0186] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0187] c. selecting patients whose ctDNA decreases by <2-log between baseline and one R-CHOP treatment cycle, and continuing treatment.R-CHOP treatment, for a total of six cycles (cycles 1 to 6), plus glimepiride, for a total of eight cycles (cycles 3 to 10). Specification 18 / 71 pages 22 CN 122070142 A

[0188] In one aspect of the invention, a third treatment cycle comprises a first dose and a second dose of glimepiride, followed by a rest period. In one such embodiment, the third treatment cycle comprises a first dose of glimepiride on day 8 of the third cycle and a second dose of glimepiride on day 15 of the third cycle, followed by a 6-day rest. In one embodiment, the subsequent two cycles (cycles 4 to 6) comprise a dose of glimepiride administered on day 8 of that cycle, followed by a 13-day rest. In one embodiment, the remaining subsequent cycles (cycles 7 to 10) comprise a dose of glimepiride administered on day 1 of that cycle, followed by a 20-day rest.

[0189] In one embodiment, a treatment cycle comprises 21 days. In another embodiment, a treatment cycle comprises 14 days. The treatment regimen according to the invention may include 6 or more treatment cycles, particularly 10 treatment cycles.

[0190] In one embodiment, glimetuzumab is administered at a dose of 2.5 mg on day 8 of cycle 3, at a dose of 10 mg on day 15 of cycle 3, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 30 mg on day 1 of cycles 7 to 10.

[0191] In another embodiment, glimetuzumab is administered at a dose of 0.5 mg on day 4 of cycle 3, at a dose of 2.5 mg on day 8 of cycle 3, at a dose of 10 mg on day 15 of cycle 3, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 30 mg on day 1 of cycles 7 to 10.

[0192] In another embodiment, glimetuzumab is administered at a dose of 0.5 mg on day 8 of cycle 3 and at a dose of 2 mg on day 9, at a dose of 10 mg on day 15 of cycle 3, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 30 mg on day 1 of cycles 7 to 10.

[0193] In one aspect of the invention, glimetuzumab is provided for use in a method of treating previously untreated diffuse large B-cell lymphoma, the method comprising:

[0194] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone.(R-CHOP) at least 2 cycles;

[0195] b. Determine the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0196] c. Select patients whose ctDNA decreases <2-log between baseline and one R-CHOP treatment cycle and continue R-CHOP treatment for a total of six cycles (cycles 1 to 6), and add glimetuzumab for a total of 8 cycles (cycles 3 to 10), wherein glimetuzumab is administered at doses of 2.5 mg and 10 mg on different days of cycle 3 and at doses of 30 mg on cycles 4 to 10.

[0197] In one aspect of the invention, glimetuzumab is provided in a method of treating previously untreated diffuse large B-cell lymphoma, the method comprising:

[0198] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for two cycles;

[0199] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0200] c. selecting patients whose ctDNA decreases <2-log between baseline and one R-CHOP treatment cycle and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of eight cycles (cycles 3 to 10), wherein glimetuzumab is administered at doses of 2.5 mg and 10 mg on different days of cycle 3 and at doses of 30 mg on cycles 4 to 10.

[0201] In one aspect of the invention, glimetuzumab is provided in a method of treating previously untreated diffuse large B-cell lymphoma, the method comprising:

[0202] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least two cycles;

[0203] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0204] c. selecting patients whose ctDNA decreased by <2-log between baseline and one R-CHOP treatment cycle, and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of eight cycles (cycles 3 to 10), wherein glimetuzumab is administered at a dose of 2.5 mg on day 8 of cycle 3 and on day 15 of cycle 3. Tianyi 10The dosage is 30 mg, and 30 mg on day 8 of cycles 4 to 6, and 30 mg on day 1 of cycles 7 to 10.

[0205] In one aspect of the invention, glimetuzumab is provided in a method of treating previously untreated diffuse large B-cell lymphoma, the method comprising:

[0206] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for two cycles;

[0207] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0208] c. selecting patients whose ctDNA decreased by <2-log between baseline and one R-CHOP treatment cycle and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of eight cycles (cycles 3 to 10), wherein glimetuzumab is administered at a dose of 2.5 mg on day 8 of cycle 3, at a dose of 10 mg on day 15 of cycle 3, and at a dose of 30 mg on day 8 of cycles 4 to 6. The dosage is mg, and a dose of 30 mg is administered on day 1 of cycles 7 to 10.

[0209] In one embodiment, glimetuzumab is administered over 4 hours (±15 minutes) on days 8 and 15 of cycle 3, day 8 of cycles 4 to 6, and day 1 of cycles 7 to 10. In another embodiment, the infusion time is extended to a maximum of 8 hours. In another embodiment, the infusion time is shortened to 2 hours (±15 minutes). In one embodiment, glimetuzumab is administered over 2 hours (±15 minutes) on day 8 of cycles 5 to 6. In one embodiment, glimetuzumab is administered over 4 hours (±15 minutes) on days 8 and 15 of cycle 3, day 8 of cycle 4, and day 8 of cycle 5 to 6. In one embodiment, glimetuzumab is administered over 4 hours (±15 minutes) on days 8 and 15 of cycle 3, day 8 of cycle 4, and over 2 hours (±15 minutes) on days 8 of cycles 5 to 6 and day 1 of cycles 7 to 10. In another embodiment, the infusion time is reduced to 90 minutes (±30 minutes). In one embodiment, glimetuzumab is administered over 90 minutes (±30 minutes) on day 1 of cycles 7, 8, 9, and 10. In one embodiment, glimetuzumab is administered over 90 minutes (±30 minutes) on day 8 of cycle 3.On days 8 and 15 of cycle 3, on day 8 of cycle 4 to 6, the drug was administered over 4 hours (±15 minutes), and on day 1 of cycles 7, 8, 9, and 10, the drug was administered over 90 minutes (±30 minutes). In one embodiment, glimetuzumab was administered on days 8 and 15 of cycle 3, on day 8 of cycle 4, over 4 hours (±15 minutes), on day 8 of cycle 5 to 6, over 2 hours (±15 minutes), and on day 1 of cycles 7, 8, 9, and 10, over 90 minutes (±30 minutes).

[0210] In any of the above aspects of the invention, ctDNA is circulating tumor DNA in plasma. In one such embodiment, the amount of ctDNA is measured in a blood sample from the patient. In one embodiment, the amount of ctDNA is measured in genomic DNA isolated from peripheral mononuclear cells or plasma-depleted whole blood (PDWB) or in cell-free DNA (cfDNA) in plasma. In one embodiment, ctDNA is measured in cell-free DNA from plasma.

[0211] In one embodiment, ctDNA is determined by targeted next-generation sequencing (NGS). In one embodiment, immunoglobulin loci (i.e., IGH, IGL, and IGK) are assessed. In one embodiment, ctDNA is determined by high-throughput amplicon sequencing (IgHTS; or clonoSEQ assay) of locus A in the immunoglobulin gene specification 20 / 71 page 24 CN 122070142. IgHTS allows for sensitive detection of lymphoma-specific clonal types from cellular DNA or cfDNA by amplifying only rearranged VDJ sequences using shared primers followed by sequencing library construction. In another embodiment, ctDNA levels are assessed using multiple genomic alterations. In one embodiment, multiple genomic alterations are defined for NHL or DLBCL. In one embodiment, ctDNA levels are assessed using a targeted ultra-deep NGS platform. These platforms detect multiple mutations in each individual case. In one embodiment, the targeted ultra-deep NGS platform is based on a hybridization capture method (CAPP-SEQ, as described by Newman AM, Bratman SV, To J et al., An ultrasensitive method for quantitating circulating tumor DNA with broad patient coverage. Nat Med. 2014;20(5):548-554, the entirety of which is incorporated herein by reference). In another embodiment, evaluation is based on an amplicon-based method.ctDNA (i.e., Lymphopanel, as described by Bohers E, Viailly PJ, Becker S et al., Non-invasive monitoring of diffuse large B-cell lymphoma by cell-free DNA high-throughput targeted sequencing: analysis of a prospective cohort. Blood Cancer J. 2018;8(8):74, which is incorporated herein by reference in its entirety). Hybrid capture-based and amplicon-based methods utilize ultra-high sequencing depths by focusing on relatively small portions of the genome. By sequencing thousands of unique molecules at each location, while utilizing PCR-induced error suppression, these methods enable sensitive detection of residual disease from plasma DNA. Monitoring ctDNA levels via targeted sequencing first requires the identification of tumor-specific alterations, typically via sequencing of tumor DNA or pre-treatment plasma DNA, as well as matching germline DNA used to remove primary variants. This yields a series of tumor-specific alterations that can then be investigated in future samples to measure ctDNA levels. By targeting multiple recurrently mutated regions of the genome, including those affected by single nucleotide variants (SNVs), insertions / deletions, gene rearrangements, and copy number variations (CNVs), these methods enable deeper insights into tumor biology and ultrasensitive disease detection. Compared to IgHTS, the initial approach using CAPP-Seq identified >100 independent mutations that enabled tumor surveillance, thereby increasing disease detection from 74% to 100% in pretreatment ctDNA samples from 23 DLBCL patients (Scherer F, Kurtz DM, Newman AM et al. Distinct biological subtypes and patterns of genome evolution in lymphoma revealed by circulating tumor DNA. Sci Transl Med. 2016;8(364):364ra155, which is incorporated herein by reference).

[0212] In one embodiment, cancer personalized deep sequencing analysis (CAPPseq) was used to determine (tumor-targeted NGS group), AVENIO NHL assay, SNV (single nucleotide variant) measurement assay, immunoglobulin (Ig) NGS, Clonoseq assay (Adaptive), and PhaseSeq.ctDNA is measured using either tumor-targeted NGS or a Foresight assay that measures PV (phase variants, multiple mutations on a single DNA molecule).

[0213] The AVENIO Oncology Assay (AOA-NHL) for non-Hodgkin lymphoma uses targeted NGS and sequencing of genomic DNA isolated from peripheral mononuclear cells or plasma-depleted whole blood (PDWB) and plasma cell-free DNA (cfDNA) to qualitatively detect ctDNA as an indicator of minimal residual disease (MRD). The AOA-NHL assay can be used to determine the ctDNA status of patients with DLBCL (Herrera et al. Blood Adv 2022, incorporated herein by reference). In one embodiment, ctDNA is determined using plasma via the AVENIO Oncology Assay (AOA-NHL) for non-Hodgkin lymphoma.

[0214] In one embodiment, the AVENIO NHL CAPP-Seq assay is used to analyze SNV mutations in ctDNA at baseline. In one embodiment, ctDNA is determined by subtracting the sample variants present in a normal PDWB sample from those variants detected in a plasma sample to identify tumor-specific variants (TSVs).

[0215] In one aspect of the invention, glimetuzumab is provided as a method for treating previously untreated diffuse large AB cell lymphoma, comprising:

[0216] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0217] b. determining the amount of ctDNA from a plasma sample on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0218] c. selecting patients whose ctDNA decreases <2-log between baseline and one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimetuzumab.

[0219] In one aspect of the invention, glimepiride is provided in a method of treating previously untreated diffuse large B-cell lymphoma, the method comprising:

[0220] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least two cycles;

[0221] b. determining the amount of ctDNA from plasma samples on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0222] c. selecting patients whose ctDNA decreased by <2-log between baseline and one R-CHOP treatment cycle, andContinue R-CHOP treatment for a total of six cycles (cycles 1 to 6), and add glimepiride for a total of eight cycles (cycles 3 to 10).

[0223] In one aspect of the invention, a method of using glimetuzumab for treating previously untreated diffuse large B-cell lymphoma is provided, the method comprising:

[0224] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0225] b. determining the amount of ctDNA from plasma samples on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0226] c. selecting patients whose ctDNA decreases <2-log between baseline and one cycle of R-CHOP treatment and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of 8 cycles (cycles 3 to 10), wherein glimetuzumab is administered at doses of 2.5 mg and 10 mg on different days of cycle 3 and at doses of 30 mg on cycles 4 to 10.

[0227] In one aspect of the invention, glimetuzumab is provided in a method of treating previously untreated diffuse large B-cell lymphoma, the method comprising:

[0228] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0229] b. determining the amount of ctDNA from plasma samples on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0230] c. selecting patients whose ctDNA decreases <2-log between baseline and one R-CHOP treatment cycle and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of 8 cycles (cycles 3 to 10), wherein glimetuzumab is administered at a dose of 2.5 mg on day 8 of cycle 3, and at a dose of 10 mg on day 15 of cycle 3, and at a dose of 10 mg on day 8 of cycles 4 to 6. Administered at a dose of 30 mg on day 1 of cycles 7 to 10.

[0231] In one aspect of the invention, glimepiride is provided in a method of treating previously untreated diffuse large B-cell lymphoma, the method comprising:

[0232] a. Administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) at least 2

[0233] b. The amount of ctDNA was determined on day 1 of cycle 1 (baseline) and day 1 of cycle 2 using plasma AVENIO oncology assay (AOA-NHL) for non-Hodgkin lymphoma; and

[0234] c. Patients with a ctDNA reduction of <2-log between baseline and one R-CHOP treatment cycle were selected and continued to be treated with R-CHOP and glimetuzumab.

[0235] In one aspect of the invention, a method of using glimetuzumab for treating previously untreated diffuse large B-cell lymphoma is provided, the method comprising:

[0236] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0237] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2 using plasma-based AVENIO oncology assay for non-Hodgkin lymphoma (AOA-NHL); and

[0238] c. selecting patients whose ctDNA decreases <2-log between baseline and one cycle of R-CHOP treatment and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of 8 cycles (cycles 3 to 10).

[0239] In one aspect of the invention, glimetuzumab is provided in a method of treating previously untreated diffuse large B-cell lymphoma, the method comprising:

[0240] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0241] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2 using plasma AVENIO oncology assay for non-Hodgkin lymphoma (AOA-NHL); and

[0242] c. selecting patients whose ctDNA decreased by <2-log between baseline and one cycle of R-CHOP treatment and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of 8 cycles (cycles 3 to 10), wherein glimetuzumab is administered at doses of 2.5 mg and 10 mg on different days of cycle 3. The dosage was administered, and at a dose of 30 mg in cycles 4 to 10.

[0243] In one embodiment of any of the foregoing aspects of the invention, rituximab was administered intravenously at a dose of 375 mg / m2. In one embodiment, rituximab was administered every 21...Rituximab was administered intravenously at a dose of 375 mg / m² every 21 days on day 1 of cycles 1 through 6, together with CHOP.

[0244] In one embodiment, CHOP administration continued for six 21-day cycles. In one embodiment, CHOP was administered as follows: cyclophosphamide was administered intravenously at a dose of 750 mg / m², doxorubicin was administered intravenously at a dose of 50 mg / m², vincristine was administered via IV bolus at a dose of 1.4 mg / m², and prednisone or prednisolone was administered at a dose of 100 mg / day. In one embodiment, cyclophosphamide is administered intravenously at a dose of 750 mg / m² on day 1, doxorubicin is administered intravenously at a dose of 50 mg / m² on day 1, vincristine is administered via IV bolus at a dose of 1.4 mg / m² on day 1, and prednisone or prednisolone is administered at a dose of 100 mg / day from day 1 to day 5. In one embodiment, the maximum dose of vincristine per cycle is 2 mg. In one embodiment, for subjects ≥70 years of age, the maximum dose of vincristine per cycle is 1.5 mg. In one embodiment, prednisone or prednisolone is administered intravenously on day 1, and the remaining doses from day 2 to day 5 are administered orally. In one embodiment, all doses of prednisone or prednisolone are administered orally. In one embodiment, prednisone or prednisolone is replaced by 80 mg of prednisolone.

[0245] In one embodiment, prednisone (100 mg) or prednisolone (100 mg) is administered prior to rituximab.

[0246] In one embodiment of any of the foregoing aspects of the invention, rituximab is administered intravenously at a dose of 375 mg / m², cyclophosphamide is administered intravenously at a dose of 750 mg / m², doxorubicin is administered intravenously at a dose of 50 mg / m², vincristine is administered via IV bolus at a dose of 1.4 mg / m², and prednisone is administered at a dose of 100 mg / day from day 1 to day 5.

[0247] In one embodiment of any of the foregoing aspects of the invention, the length of each cycle is 21 days.

[0248] In one embodiment of any of the foregoing aspects of the invention, administration of such treatment to multiple human patients at high risk of ctDNA produces an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%. In one embodiment, the ORR is at least 90%. In one embodiment, the ORR is at least 95%.

[0249] In one embodiment, ORR is determined at the end of treatment. In one embodiment, ORR is determined by PET-CT scan. In one embodiment, ORR is determined according to the 2014 Lugano response criteria. In one embodiment, ORR is determined between day 15 and day 21 of cycle 2. In one embodiment, ORR is measured by PET / CT scan and determined at the end of treatment by an automated response (aLugano) according to the 2014 Lugano response criteria. In one embodiment, aLugano analysis is performed in a PET / CT-evaluable population defined as all patients with available PET / CT scans at baseline and during treatment.

[0250] In one embodiment of any of the foregoing aspects of the invention, administration of such treatment to multiple human patients at high risk of ctDNA produces a complete response (CR) rate of at least 75%, at least 80%, or at least 85%. In one embodiment, the CR rate is at least 80%. In one embodiment, the CR rate is at least 85%.

[0251] In one embodiment, a method of using glimetuzumab for treating previously untreated diffuse large B-cell lymphoma at high risk of ctDNA includes administering glimetuzumab to a patient in combination with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), wherein the patient is identified as at high risk of ctDNA if the ctDNA decreases by less than 2-log after one R-CHOP cycle, wherein administration of such treatment to multiple human patients at high risk of ctDNA produces an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%. In one embodiment, the ORR is at least 90%. In another embodiment, the ORR is at least 95%.

[0252] In one embodiment, glimetuzumab is provided in a method of treating previously untreated diffuse large B-cell lymphoma at high risk of ctDNA, the method comprising administering glimetuzumab to a patient in combination with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), wherein the patient is identified as at high risk of ctDNA if the ctDNA decreases by less than 2-log after one R-CHOP cycle, wherein administration of such treatment to multiple human patients at high risk of ctDNA produces a complete response (CR) rate of at least 75%, at least 80%, or at least 85%. In one embodiment, the CR rate is at least 80%. In another embodiment, the CR rate is at least 85%.

[0253] In one embodiment, ctDNA reduction is determined by measuring ctDNA levels before or on the day of the first treatment (baseline) and after at least one R-CHOP treatment cycle. In one embodiment, ctDNA levels are determined on or before day 1 of the first R-CHOP treatment cycle (baseline) and day 1 of the second R-CHOP treatment cycle. In another embodiment, ctDNA levels are determined on day 1 of the first R-CHOP treatment cycle (baseline) and day 1 of the second R-CHOP treatment cycle. In one embodiment, a patient is identified as high-risk for ctDNA if the reduction in ctDNA is less than 2 log. In one embodiment, a patient is identified as high-risk for ctDNA if the reduction in ctDNA levels between day 1 of cycle 1 and day 1 of cycle 2, as determined by the AOA-NHL test, is less than 2 log-fold.

[0254] One aspect of the invention relates to glimetuzumab for the treatment of previously untreated diffuse large B-cell lymphoma at high (ct)DNA risk, the method comprising administering glimetuzumab to a patient in combination with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), wherein the patient is identified as ctDNA high-risk if the patient's ctDNA reduction is less than 2-log, wherein the ctDNA reduction is measured on or before day 1 of the first R-CHOP treatment cycle (baseline) and day 1 of the second R-CHOP treatment cycle, wherein administration of such treatment to multiple ctDNA high-risk human patients produces an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%. In one embodiment, the ORR is at least 90%. In another embodiment, the ORR is at least 95%. In one such embodiment, the reduction in ctDNA was measured on day 1 of the first R-CHOP treatment cycle (baseline) and day 1 of the second R-CHOP treatment cycle.

[0255] One aspect of the invention relates to glimetuzumab in a method of treating previously untreated diffuse large B-cell lymphoma with high (ct)DNA risk, the method comprising administering glimetuzumab to a patient in combination with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), wherein the patient has been identified as high-risk for ctDNA if the reduction in ctDNA is less than 2-log, wherein the reduction in ctDNA is measured on day 1 of the first R-CHOP treatment cycle (baseline).Measurements were taken on day 1 of the first R-CHOP treatment cycle (baseline) and day 1 of the second R-CHOP treatment cycle, wherein administration of such treatment to multiple human patients at high risk of ctDNA resulted in a complete response (CR) rate of at least 75%, at least 80%, or at least 85%. In one embodiment, the CR rate was at least 80%. In another embodiment, the CR rate was at least 85%.

[0256] In one such embodiment, the reduction in ctDNA was measured on day 1 of the first R-CHOP treatment cycle (baseline) and day 1 of the second R-CHOP treatment cycle.

[0257] One aspect of the invention relates to glimetuzumab for treating previously untreated diffuse large B-cell lymphoma to produce a complete response (CR) rate of at least 75%, at least 80%, or at least 85%, the method comprising:

[0258] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0259] b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0260] c. selecting patients with a ctDNA reduction of <2.5-log between baseline and at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimetuzumab, wherein

[0261] in one embodiment, the CR rate is at least 80%. In one embodiment, the CR rate is at least 85%.

[0262] One aspect of the invention relates to glimetuzumab for treating previously untreated diffuse large B-cell lymphoma to produce an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%, the method comprising:

[0263] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0264] b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0265] c. selecting patients with a ctDNA reduction of <2.5 log between baseline and at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimetuzumab, wherein

[0266] in one embodiment, the ORR is at least 90%. In one embodiment, the ORR is at least 95%. Specification 25 / 71 pages 29 CN 122070142 A

[0267] One aspect of the invention relates to glimetuzumab for the treatment of previously untreated patients with high (ct)DNA risk.A method for treating diffuse large B-cell lymphoma to produce a complete response (CR) rate of at least 75%, at least 80%, or at least 85%, comprising:

[0268] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0269] b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0270] c. selecting patients with a ctDNA reduction of <2.5-log between baseline and at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimepiride.

[0271] In one embodiment, the CR rate is at least 80%. In one embodiment, the CR rate is at least 85%.

[0272] In another aspect of the invention, glimetuzumab is provided in a method of treating previously untreated diffuse large B-cell lymphoma to produce a complete response (CR) rate of at least 75%, at least 80%, or at least 85%, the method comprising:

[0273] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0274] b. determining the amount of ctDNA after cycle 1 (baseline) and at least one R-CHOP treatment cycle; and

[0275] c. selecting patients whose ctDNA reduction is <2.5-log between baseline and at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimetuzumab.

[0276] In one embodiment, the CR rate is at least 80%. In one embodiment, the CR rate is at least 85%.

[0277] In one embodiment, step a) includes administering R-CHOP on day 1 of cycles 1, 2, and optionally cycle 3. In one embodiment, step a) includes administering R-CHOP on day 1 of cycles 1 and 2.

[0278] In one embodiment, step b) includes determining the amount of ctDNA on day 1 or before (baseline) of cycle 1 of R-CHOP treatment in step a) and on day 1 of cycle 2 or day 1 of cycle 3. In one embodiment, step b) includes determining the amount of ctDNA on day 1 (baseline) of cycle 1 of R-CHOP treatment in step a) and on day 1 of cycle 2.

[0279] In one embodiment, step c) includes selecting patients with a ctDNA reduction of <2-log. In one embodiment, step c) includes continuing R-CHOP treatment.Treatment was continued for a total of six cycles (cycles 1 to 6), with the addition of glimetuzumab for a total of eight cycles (cycles 3 to 10).

[0280] In one aspect of the invention, glimetuzumab is provided in a method of treating previously untreated diffuse large B-cell lymphoma to produce a complete response (CR) rate of at least 75%, at least 80%, or at least 85%, comprising:

[0281] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least two cycles;

[0282] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0283] c. selecting patients whose ctDNA decreased by <2-log between baseline and one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimetuzumab.

[0284] In one embodiment, the CR rate is at least 80%. In one embodiment, the CR rate is at least 85%.

[0285] In another aspect of the invention, glimetuzumab is provided in a method for treating previously untreated diffuse large B-cell lymphoma to produce a complete response (CR) rate of at least 75%, at least 80%, or at least 85%, as described in page 26 / 71 of this specification, CN 122070142 A. The method comprises:

[0286] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least two cycles;

[0287] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0288] c. selecting patients whose ctDNA decreases by <2-log between baseline and one R-CHOP treatment cycle and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of eight cycles (cycles 3 to 10).

[0289] In one embodiment, the CR rate is at least 80%. In one embodiment, the CR rate is at least 85%.

[0290] In another aspect of the invention, glimepiride is provided in a method of treating previously untreated diffuse large B-cell lymphoma to produce a complete response (CR) rate of at least 75%, at least 80%, or at least 85%, comprising:

[0291] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for two cycles;

[0292] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0293] c. Patients with a ctDNA reduction of <2-log between baseline and one R-CHOP treatment cycle are selected and treated with R-CHOP for a total of six cycles (cycles 1 to 6), with the addition of glimetuzumab for a total of eight cycles (cycles 3 to 10).

[0294] In one embodiment, the CR rate is at least 80%. In one embodiment, the CR rate is at least 85%.

[0295] In one aspect of the invention, the third treatment cycle comprises a first dose and a second dose of glimetuzumab, followed by a rest period. In one such embodiment, the third treatment cycle comprises a first dose of glimetuzumab on day 8 of the third cycle and a second dose of glimetuzumab on day 15 of the third cycle, followed by a 6-day rest period. In one embodiment, the subsequent two cycles (cycles 4 to 6) comprise a dose of glimetuzumab administered on day 8 of the cycle, followed by a 13-day rest period. In one embodiment, the remaining subsequent cycles (cycles 7 to 10) comprise a dose of glimetuzumab administered on day 1 of the cycle, followed by a 20-day rest period.

[0296] In one embodiment, a treatment cycle comprises 21 days. In another embodiment, a treatment cycle comprises 14 days. Treatment regimens according to the invention may comprise 6 or more treatment cycles, particularly 10 treatment cycles.

[0297] In one embodiment, glimetuzumab is administered at a dose of 2.5 mg on day 8 of cycle 3, at a dose of 10 mg on day 15 of cycle 3, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 30 mg on day 1 of cycles 7 to 10.

[0298] In another embodiment, glimetuzumab is administered at a dose of 0.5 mg on day 4 of cycle 3, at a dose of 2.5 mg on day 8 of cycle 3, at a dose of 10 mg on day 15 of cycle 3, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 30 mg on day 1 of cycles 7 to 10.

[0299] In another embodiment, glimepiride is administered at a dose of 0.5 mg on day 8 of cycle 3, a dose of 2 mg on day 9, a dose of 10 mg on day 15 of cycle 3, a dose of 30 mg on day 8 of cycles 4 through 6, and a dose of 30 mg on day 1 of cycles 7 through 10.

[0300] In one aspect of the invention, glimetuzumab is provided for use in the treatment of previously untreated diffuse large B-cell lymphoma to produce a complete response (CR) rate of at least 75%, at least 80%, or at least 85%, as described in page 27 / 71 of CN 122070142 A. The method comprises:

[0301] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least two cycles;

[0302] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0303] c. selecting patients whose ctDNA decreases <2-log between baseline and one R-CHOP treatment cycle and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of eight cycles (cycles 3 to 10), wherein glimetuzumab is added in cycle 3. The drug was administered at doses of 2.5 mg and 10 mg on different days, and at a dose of 30 mg on cycles 4 to 10.

[0304] In one embodiment, the CR rate was at least 80%. In another embodiment, the CR rate was at least 85%.

[0305] In one aspect of the invention, glimetuzumab is provided in a method of treating previously untreated diffuse large B-cell lymphoma to produce a complete response (CR) rate of at least 75%, at least 80%, or at least 85%, the method comprising:

[0306] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for two cycles;

[0307] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0308] c. selecting patients whose ctDNA decreases <2-log between baseline and one R-CHOP treatment cycle and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of eight cycles (cycles 3 to 10), wherein glimetuzumab is administered at doses of 2.5 mg and 10 mg on different days of cycle 3. The dosage is administered, and in cycles 4 to 10 at a dose of 30 mg.

[0309] In one embodiment, the CR rate is at least 80%. In one embodiment, the CR rate is at least 85%.

[0310] In one aspect of the invention, glimepiride is provided for the treatment of previously untreated diffuse large B-cell lymphoma to produce at least 75%, at least 80%, or at leastThe method for achieving an 85% complete response (CR) rate includes:

[0311] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least two cycles;

[0312] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0313] c. selecting patients whose ctDNA decreased by <2-log between baseline and one R-CHOP treatment cycle and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of eight cycles (cycles 3 to 10), wherein glimetuzumab is administered at a dose of 2.5 mg on day 8 of cycle 3, at a dose of 10 mg on day 15 of cycle 3, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 30 mg on day 1 of cycles 7 to 10.

[0314] In one embodiment, the CR rate is at least 80%. In one embodiment, the CR rate is at least 85%.

[0315] In one aspect of the invention, glimetuzumab is provided in a method of treating previously untreated diffuse large B-cell lymphoma to produce a complete response (CR) rate of at least 75%, at least 80%, or at least 85%, comprising:

[0316] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for two cycles;

[0317] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0318] c. selecting patients whose ctDNA decreases <2-log between baseline and one R-CHOP treatment cycle, and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of eight cycles (cycle 3 to 6). (to 10), wherein glimetuzumab is administered at a dose of 2.5 mg on day 8 of cycle 3, at a dose of 10 mg on day 15 of cycle 3, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 30 mg on day 1 of cycles 7 to 10.

[0319] In one embodiment, the CR rate is at least 80%. In one embodiment, the CR rate is at least 85%.

[0320] One aspect of the invention relates to glimetuzumab for the treatment of (ct)DNAA method for producing an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95% in high-risk, previously untreated diffuse large B-cell lymphoma, comprising:

[0321] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0322] b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0323] c. selecting patients with a ctDNA reduction of <2.5-log between baseline and at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimepiride.

[0324] In one embodiment, the ORR is at least 90%. In another embodiment, the ORR is at least 95%.

[0325] In another aspect of the invention, a method of using rituximab for treating previously untreated diffuse large B-cell lymphoma to produce an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%, comprising:

[0326] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least two cycles;

[0327] b. determining the amount of ctDNA after cycle 1 (baseline) and at least one R-CHOP treatment cycle; and

[0328] c. selecting patients whose ctDNA reduction is <2.5-log between baseline and at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and rituximab.

[0329] In one embodiment, the ORR is at least 90%. In one embodiment, the ORR is at least 95%.

[0330] In one embodiment, step a) includes administering R-CHOP on day 1 of cycles 1, 2, and optionally cycle 3. In one embodiment, step a) includes administering R-CHOP on day 1 of cycles 1 and 2.

[0331] In one embodiment, step b) includes determining the amount of ctDNA on day 1 or before (baseline) of cycle 1 of R-CHOP treatment in step a) and on day 1 of cycle 2 or day 1 of cycle 3. In one embodiment, step b) includes determining the amount of ctDNA on day 1 (baseline) of R-CHOP treatment in step a) and on day 1 of cycle 2.

[0332] In one embodiment, step c) includes selecting patients with a ctDNA reduction of <2-log. In one embodimentIn step c), R-CHOP treatment is continued for a total of six cycles (cycles 1 to 6), and glimetuzumab is added for a total of eight cycles (cycles 3 to 10).

[0333] In one aspect of the invention, a method is provided for treating previously untreated diffuse large B-cell lymphoma with rituximab to produce an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%, comprising:

[0334] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least two cycles;

[0335] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and specification 29 / 71 pages 33 CN 122070142 A

[0336] c. selecting patients whose ctDNA decreases by <2-log between baseline and one R-CHOP treatment cycle and continuing treatment with R-CHOP and rituximab.

[0337] In one embodiment, the ORR is at least 90%. In one embodiment, the ORR is at least 95%.

[0338] In another aspect of the invention, a method is provided for using rituximab to treat previously untreated diffuse large B-cell lymphoma to produce an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%, comprising:

[0339] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least two cycles;

[0340] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0341] c. selecting patients whose ctDNA decreases <2-log between baseline and one R-CHOP treatment cycle and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding rituximab for a total of eight cycles (cycles 3 to 10).

[0342] In one embodiment, the ORR is at least 90%. In one embodiment, the ORR is at least 95%.

[0343] In another aspect of the invention, a method of using rituximab to treat previously untreated diffuse large B-cell lymphoma to produce an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95% is provided, the method comprising:

[0344] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for 2 cycles;

[0345] b. Determine the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0346] c. Select patients whose ctDNA decreases by <2-log between baseline and one R-CHOP treatment cycle and continue R-CHOP treatment for a total of six cycles (cycles 1 to 6), and add glimetuzumab for a total of eight cycles (cycles 3 to 10).

[0347] In one embodiment, the ORR is at least 90%. In one embodiment, the ORR is at least 95%.

[0348] In one aspect of the invention, the third treatment cycle comprises a first dose and a second dose of glimetuzumab, followed by a rest period. In one such embodiment, the third treatment cycle comprises a first dose of glimetuzumab on day 8 of the third cycle and a second dose of glimetuzumab on day 15 of the third cycle, followed by a 6-day rest period. In one embodiment, the subsequent two cycles (cycles 4 to 6) comprise a dose of glimetuzumab administered on day 8 of that cycle, followed by a 13-day rest period. In one embodiment, the remaining subsequent cycles (cycles 7 to 10) consist of a dose of glimetuzumab administered on day 1 of that cycle, followed by a 20-day rest period.

[0349] In one embodiment, a treatment cycle comprises 21 days. In another embodiment, a treatment cycle comprises 14 days. Treatment regimens according to the invention may comprise 6 or more treatment cycles, particularly 10 treatment cycles.

[0350] In one embodiment, glimetuzumab is administered at a dose of 2.5 mg on day 8 of cycle 3, at a dose of 10 mg on day 15 of cycle 3, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 30 mg on day 1 of cycles 7 to 10.

[0351] In another embodiment, glimetuzumab is administered at a dose of 0.5 mg on day 4 of cycle 3, at a dose of 2.5 mg on day 8 of cycle 3, at a dose of 10 mg on day 15 of cycle 3, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 30 mg on day 1 of cycles 7 to 10. Specification 30 / 71 pages 34 CN 122070142 A

[0352] In another embodiment, glimetuzumab is administered at a dose of 0.5 mg on day 8 of cycle 3, at a dose of 2 mg on day 9, at a dose of 10 mg on day 15 of cycle 3, and at a dose of 30 mg on day 8 of cycles 4 to 6.The dose is 30 mg, and it is also administered on day 1 of cycle 7 to 10.

[0353] In one aspect of the invention, glimetuzumab is provided for use in the treatment of previously untreated diffuse large B-cell lymphoma to produce an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%, the method comprising:

[0354] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0355] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0356] c. selecting patients whose ctDNA decreases <2-log between baseline and one R-CHOP treatment cycle and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of 8 cycles (cycles 3 to 10), wherein glimetuzumab is administered at 2.5 mg on different days of cycle 3. The dose is administered at a dose of 10 mg, and at a dose of 30 mg in cycles 4 to 10.

[0357] In one embodiment, the ORR is at least 90%. In another embodiment, the ORR is at least 95%.

[0358] In one aspect of the invention, glimetuzumab is provided in a method of treating previously untreated diffuse large B-cell lymphoma to produce an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%, comprising:

[0359] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for two cycles;

[0360] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0361] c. selecting patients whose ctDNA decreases <2-log between baseline and one R-CHOP treatment cycle and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of eight cycles (cycles 3 to 10), wherein glimetuzumab is administered at a dose of 2.5 mg on different days of cycle 3 and 10 Administered at a dose of mg, and at a dose of 30 mg in cycles 4 to 10.

[0362] In one embodiment, the ORR is at least 90%. In one embodiment, the ORR is at least 95%.

[0363] In one aspect of the invention, glimepiride is provided for the treatment of previously untreated patients.A method for treating diffuse large B-cell lymphoma to produce an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%, comprising:

[0364] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0365] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0366] c. selecting patients whose ctDNA decreased by <2-log between baseline and one cycle of R-CHOP treatment and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of 8 cycles (cycles 3 to 10), wherein glimetuzumab is administered at a dose of 2.5 mg on day 8 of cycle 3, and at a dose of 10 mg on day 15 of cycle 3, and ...10 mg on day 10 of cycle 3, and at a dose of 2.5 mg on day 8 of cycle 3, and at a dose of 10 mg on day 10 of cycle 3, and at a dose of 10 mg on day 10 of cycle 3, and at a dose of 10 mg on day 10 of cycle 3, and The drug is administered at a dose of 30 mg on day 8 and at a dose of 30 mg on day 1 of cycles 7 through 10.

[0367] In one embodiment, the ORR is at least 90%. In another embodiment, the ORR is at least 95%.

[0368] In one aspect of the invention, glimetuzumab is provided for use in the treatment of previously untreated diffuse large B-cell lymphoma to produce an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%, as described in the specification on page 31 / 71 of CN 122070142 A, comprising:

[0369] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for two cycles;

[0370] b. determining the amount of ctDNA on day 1 of cycle 1 (baseline) and day 1 of cycle 2; and

[0371] c. selecting patients whose ctDNA decreases <2-log between baseline and one R-CHOP treatment cycle and continuing R-CHOP treatment for a total of six cycles (cycles 1 to 6), and adding glimetuzumab for a total of eight cycles (cycle 3 to 6). (to 10), wherein glibenclamide is administered at a dose of 2.5 mg on day 8 of cycle 3, at a dose of 10 mg on day 15 of cycle 3, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 30 mg on day 1 of cycles 7 to 10.

[0372] In one embodiment, the ORR is at least90%. In one embodiment, the ORR is at least 95%.

[0373] In one embodiment, the CR rate is determined at the end of treatment. In one embodiment, the CR rate is determined by PET-CT scan. In one embodiment, the CR rate is determined according to the 2014 Lugano Response Criteria. In one embodiment, the CR rate is determined between day 15 and day 21 of cycle 2. In one embodiment, the CR rate is measured by PET / CT scan and determined at the end of treatment by an automated response (aLugano) according to the 2014 Lugano Response Criteria. In one embodiment, the aLugano analysis is performed in a PET / CT-evaluable population, defined as all patients with available PET / CT scans at baseline and during treatment.

[0374] In one embodiment, tumor assessment is performed before the first treatment (on or before day 1 of cycle 1), during intermediate response assessment (between day 15 and day 21 of cycle 2), and at EOT using 18FFDG-PET / CT and diagnostic CT scans.

[0375] In one embodiment, the presence and extent of lymphadenopathy, hepatomegaly, and splenomegaly are determined by physical examination. In one embodiment, the response is assessed based on the results of physical examination, CT scan, and FDG-PET / CT scan.

[0376] In one embodiment of any of the foregoing aspects of the invention, administration of such treatment to multiple human patients at high risk of ctDNA results in an improved duration of response (DOR) rate compared to R-CHOP treatment alone. In one embodiment, the DOR rate is determined according to the 2014 Lugano Response Criteria. In one embodiment, the DOR rate is measured by PET / CT scan and determined by an automated response (aLugano) according to the 2014 Lugano Response Criteria. In one embodiment, the aLugano analysis is performed in a PET / CT-evaluable population defined as all patients with available PET / CT scans at baseline and during treatment. In one embodiment, DOR is defined as the time from the first occurrence of a documented objective response (CR or PR) to disease progression, relapse, or death from any cause, whichever occurs first. In one embodiment, the DOR analysis includes only participants with an objective response (CR or PR).

[0377] In one embodiment of any of the foregoing aspects of the invention, administration of such treatment to multiple human patients at high risk of ctDNA resulted in improved progression-free survival (PFS) compared to R-CHOP treatment alone.

[0378] In one embodiment of any of the foregoing aspects of the invention, administration of such treatment to multiple human patients at high risk of ctDNA resulted in improved overall survival (OS) compared to R-CHOP treatment alone.

[0379] IPI is widely used to stratify patients with DLBCL based on their age, biochemical parameters, performance status, and disease stage. In one embodiment, the performance status of the subjects to be treated was 0-2 in the Eastern Cooperative Oncology Group (ECOG, see Oken MM, Creech RH, Tormey DC et al. Toxicity and response criteria of the Eastern Cooperative Oncology Group. Am J Clin Oncol 1982;5:649-55.). The International Prognostic Index (IPI, Shipp MA, Harrington DP, Anderson JR et al. A predictive model for aggressive non-Hodgkin's Lymphoma. N Engl J Med 1993;329:987-94.) is widely used for risk stratification of patients with DLBCL based on their age, biochemical parameters, performance status, and disease stage, as described on pages 32 / 71 of the specification (CN 122070142 A). In one embodiment, the IPI of the subject to be treated is 1-5 or 2-5. In another embodiment, the IP of the subject to be treated is 0, accompanied by a large lesion (> 6 cm). In yet another embodiment, the subject to be treated has at least one two-dimensional FDG-avid measurable lymphoma lesion on a PET / CT scan, defined as having a longest diameter > 1.5 cm on a CT scan.

[0380] In one aspect, previously untreated DLBCL is previously untreated CD-20 positive diffuse large B-cell lymphoma (DLBCL). In one embodiment, previously untreated DLBCL includes nonspecific (NOS) type DLBCL, germinal center B-cell-like (GCB) and activated B-cell-like (ABC) / non-GCB types, as well as dual-expressing lymphoma (MYC and BCL2 co-expression), high-grade B-cell lymphoma (HGBCL) with MYC and BCL2 and / or BCL6 translocations, high-grade B-cell lymphoma (HGBCL; e.g., HGBCL NOS, double-hit HGBCL, and triple-hit HGBCL), and new-onset transformed follicular lymphoma.

[0381] In one aspect, previously untreatedDLBCL excludes unclassifiable B-cell lymphomas (gray zone lymphomas), primary mediastinal (thymic) large B-cell lymphomas, T-cell / histiocytic abundant large B-cell lymphomas, Burkitt lymphomas, CNS lymphomas (primary or secondary involvement), primary exudative DLBCL, and primary cutaneous DLBCL.

[0382] In some embodiments, the method and use characterized by the present invention further includes administering one or more additional therapeutic agents to a subject. In some embodiments, the method characterized by the present invention further includes administering one or more additional therapeutic agents to a subject. In some embodiments, the one or more additional therapeutic agents are tocilizumab. In some embodiments, the one or more additional therapeutic agents are corticosteroids. In some embodiments, corticosteroids include prednisone, prednisolone, methylprednisolone, or dexamethasone. In some embodiments, the one or more additional therapeutic agents are antihistamines. In some embodiments, the antihistamine is diphenhydramine. In some embodiments, the one or more additional therapeutic agents include allopurinol and raburicase. In some embodiments, one or more additional therapeutic agents are antipyretics. In some embodiments, one or more additional therapeutic agents include granulocyte colony-stimulating factor (G-CSF). In some embodiments, one or more additional therapeutic agents are mesna.

[0383] In some embodiments, one or more additional therapeutic agents are corticosteroids. In some embodiments, corticosteroids include prednisone, prednisolone, methylprednisolone, or dexamethasone. In one embodiment, dexamethasone is administered intravenously at a dose of about 20 mg (e.g., 20 mg ± 0.1 mg, ± 0.25 mg, ± 0.5 mg, ± 1 mg, ± 1.5 mg, or ± 2 mg) at least one hour (i.e., at least one hour ± 6 minutes; for example, at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 36, 48 hours or more) before administration of any dose of the bispecific antibody (i.e., glimetuzumab). In one embodiment, dexamethasone is administered at a dose of 20 mg before any dose of glimetuzumab. In one embodiment, dexamethasone is administered at a dose of 20 mg one hour before any dose of glimetuzumab. In one embodiment, dexamethasone is administered at a dose of 20 mg before the first and second doses of glimetuzumab. In one embodiment, dexamethasone is administered at a dose of 20 mg one hour before the first and second doses of glimetuzumab. In one embodiment, the dexamethasone pre-dose is administered only during cycle 3 (i.e., the first treatment cycle using glimetuzumab). In one embodiment, at R-CHOP...Dexamethasone should be administered intravenously at a dose of about 20 mg (e.g., 20 mg ± 0.1 mg, ± 0.25 mg, ± 0.5 mg, ± 1 mg, ± 1.5 mg, or ± 2 mg) at least one hour (i.e., at least one hour ± 6 minutes; for example, at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 36, 48 hours or more) before any dose is administered. In one embodiment, methylprednisolone is administered intravenously at a dose of about 80 mg (e.g., 80 mg ± 0.5 mg, ± 1 mg, ± 1.5 mg, ± 2 mg, ± 4 mg, ± 6 mg, or ± 8 mg) at least one hour (i.e., at least one hour ± 6 minutes; for example, at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 36, 48 hours or more) before any dose of the bispecific antibody (i.e., glucuronidone). In one embodiment, methylprednisolone is administered intravenously at a dose of about 80 mg (e.g., 80 mg ± 0.5 mg, ± 1 mg, ± 1.5 mg, ± 2 mg, ± 4 mg, ± 6 mg, or ± 8 mg) at least about one hour (i.e., at least one hour ± 6 minutes; for example, at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 36, 48 hours or more) before any dose of R-CHOP is administered. In one embodiment, prednisone is administered orally at a dose of about 100 mg (e.g., 100 mg ± 0.5 mg, ± 1 mg, ± 1.5 mg, ± 2 mg, ± 4 mg, ± 6 mg, ± 6 mg, ± 8 mg, or ± 10 mg) at least one hour (i.e., at least one hour ± 6 minutes; for example, at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 36, 48 hours or more) before any dose of the bispecific antibody (i.e., glimetuzumab). In one embodiment, prednisolone is administered intravenously at a dose of about 100 mg (e.g., 100 mg ± 0.5 mg, ± 1 mg, ± 1.5 mg, ± 2 mg, ± 4 mg, ± 6 mg, ± 8 mg, or ± 10 mg) at least one hour (i.e., at least one hour ± 6 minutes; for example, at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 36, 48 hours or more) before any dose of the bispecific antibody (i.e., glimetuzumab).

[0384] In some embodiments, one or more additional therapeutic agents are antihistamines. In some embodiments, the antihistamine is diphenhydramine. In one embodiment, diphenhydramine is administered orally or intravenously at a dose of about 50 mg (e.g., 50 mg ± 0.5 mg, ± 1 mg, ± 1.5 mg, ± 2 mg, ± 3 mg, ± 4 mg, or ± 5 mg) at least 30 minutes (i.e., at least 30 minutes ± 3 minutes; for example, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 36, 48 hours or more) before administration of any dose of the bispecific antibody (i.e., glimetuzumab). In one embodiment, diphenhydramine is administered at a dose of 50 mg before administration of any dose of glimetuzumab. In one embodiment, diphenhydramine is administered at a dose of 50 mg about 30 minutes before administration of any dose of glimetuzumab.

[0385] In some embodiments, one or more additional therapeutic agents are antipyretics. In one embodiment, the antipyretic is acetaminophen or paracetamol. In one embodiment, acetaminophen or paracetamol is administered orally at a dose between about 500 mg and about 1000 mg (e.g., at least 30 minutes ± 3 minutes; for example, at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 36, 48 hours or more) before administration of any dose of the bispecific antibody (i.e., glucuronidumab). In one embodiment, acetaminophen or paracetamol is administered orally at a dose between about 500 mg and about 1000 mg (e.g., at least 30 minutes ± 3 minutes; for example, at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 36, 48 hours or more) before any dose of R-CHOP. In one embodiment, acetaminophen or paracetamol is administered at a dose of 500 or 1000 mg before any dose of glimepiride. In one embodiment, acetaminophen or paracetamol is administered at a dose of 500 or 1000 mg about 30 minutes before any dose of glimepiride.

[0386] In one embodiment, one or more additional therapeutic agents are tocilizumab. In one embodiment, tocilizumab is administered at a dose of about 8 mg / kg (e.g., 8 mg / kg ± 0.05 mg / kg, ± 0.1 mg / kg, ± 0.25 mg / kg, ± 0.5 mg / kg, or ± 0.8 mg / kg), or if the subject weighs less than 30 kg, tocilizumab is administered at a dose of about 12 mg / kg (e.g., 12 mg / kg ± 0.05 mg / kg, ± 0.1 mg / kg, ± 0.25 mg / kg, ± 0.5 mg / kg, ± 0.75 mg / kg, ± 1 mg / kg, or ± 1.2 mg / kg), and wherein the maximum dose is about 800 mg (e.g., 800 mg ± 10 mg, ± 25 mg, ± 50 mg, or ± 80 mg).

[0387] In some embodiments, one or more additional therapeutic agents are methotrexate. In one embodiment, intrathecal methotrexate is administered as CNS prophylaxis. In one embodiment, intrathecal methotrexate is administered to a subject who has four or more of the following risk factors: age > 60 years, stage III or IV disease, more than two extranodal sites, longest diameter greater than the upper limit of normal, or kidney or adrenal involvement.

[0388] In one embodiment, one or more additional therapeutic agents comprise granulocyte colony-stimulating factor (G-CSF). In one embodiment, G-CSF is administered during cycles 1 to 6. In one embodiment, G-CSF is administered during cycles 3 to 6. In one embodiment, G-CSF is filgrastim. In one embodiment, filgrastim is administered subcutaneously once daily at a dose of about 5 µg / kg up to 48 hours after administration of R-CHOP. In one embodiment, the dose of filgrastim is rounded to 300 µg or 400 µg. In one embodiment, treatment with filgrastim lasts for 7 days. In one embodiment, treatment with filgrastim continues until ANC > 1.5 × 10⁹ / L for two consecutive days. In one embodiment, filgrastim is administered subcutaneously once daily at a dose of about 5 µg / kg up to 48 hours after administration of R-CHOP. In one embodiment, filgrastim is administered on day 1 of cycles 1 to 6.

[0389] In one embodiment, G-CSF is glycolated filgrastim. In one embodiment, glycolated filgrastim is administered subcutaneously at a dose of about 6 mg 48 hours after administration of R-CHOP. In one embodiment, glycolated filgrastim is administered on day 1 of cycles 1 to 6.Administered on day 1.

[0390] In another embodiment, G-CSF is levofloxacin.

[0391] In one embodiment, G-CSF is administered approximately one to approximately two days after administration of any dose of R-CHOP (e.g., 24, 26, 28, 30, 32, 36, 38, 40, 42, 44, 46, or 48 hours). In one embodiment, filgrastim is administered intravenously or subcutaneously at a dose of about 5 µg / kg / day (e.g., 5 µg / kg / day ± 0.05 µg / kg / day, ± 0.1 µg / kg / day, ± 0.2 µg / kg / day, ± 0.3 µg / kg / day, ± 0.4 µg / kg / day, ± 0.5 µg / kg / day) or about 10 µg / kg / day (e.g., 10 µg / kg / day ± 0.1 µg / kg / day, ± 0.2 µg / kg / day, ± 0.4 µg / kg / day, ± 0.6 µg / kg / day, ± 0.8 µg / kg / day, ± 1 µg / kg / day). In one embodiment, filgrastim is administered at a dose of about 5 µg / kg / day in the first dosing cycle (e.g., 5 µg / kg / day ± 0.05 µg / kg / day, ± 0.1 µg / kg / day, ± 0.2 µg / kg / day, ± 0.3 µg / kg / day, ± 0.4 µg / kg / day, ± 0.5 µg / kg / day) in the second dosing cycle and / or each additional dosing cycle at a dose of about 10 µg / kg / day (e.g., 10 µg / kg / day ± 0.1 µg / kg / day, ± 0.2 µg / kg / day, ± 0.4 µg / kg / day, ± 0.6 µg / kg / day, ± 0.8 µg / kg / day, ± 1 µg / kg / day).

[0392] One aspect of the invention relates to the use of a bispecific antibody targeting CD3 and CD20 comprising the VH and VL sequences of glimetuzumab in the manufacture of a medicament for treating a subject with previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, wherein the patient is administered a combination of glimetuzumab and chemotherapy, wherein the patient has been identified as high-risk for ctDNA if the patient's ctDNA decreases by less than 2.5 log after one cycle of chemotherapy.

[0393] In one embodiment, the patient's ctDNA decreases by less than 2 log.

[0394] In one embodiment, the use of glimetuzumab in the manufacture of a medicament for treating a subject with previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA is provided, wherein the patient is administered glimetuzumab...The combination of monoclonal antibody and chemotherapy, wherein if a patient has been identified as high-risk for ctDNA after one cycle of chemotherapy, the patient is considered to have a lower ctDNA level.

[0395] One aspect of the invention relates to the use of glimetuzumab in the manufacture of a medicament for treating a subject with previously untreated diffuse large B-cell lymphoma at high-risk (ct)DNA, wherein the patient is administered a combination of glimetuzumab with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), wherein if the patient has been identified as high-risk for ctDNA after one R-CHOP cycle, the patient is considered to have a lower ctDNA level.

[0396] One aspect of the invention relates to the use of glimetuzumab according to any of the above embodiments in the manufacture of a medicament for treating a subject with previously untreated diffuse large B-cell lymphoma at high-risk (ct)DNA.

[0397] One aspect of the invention relates to the use of glimetuzumab in the manufacture of a medicament for treating a subject with previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, comprising:

[0398] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0399] b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0400] c. selecting patients with a ctDNA reduction of <2.5-log between baseline and at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimetuzumab.

[0401] Another aspect of the invention relates to the invention as described herein.

[0402] Each embodiment may be incorporated unless the context clearly indicates otherwise. Each embodiment may be applied to various aspects of the invention unless the context clearly indicates otherwise.

[0403] Certain embodiments of the invention will become apparent from the following more detailed description of some preferred embodiments and claims.

[0404] (iv) CRS Risk Mitigation Strategy

[0405] The present invention relates to a method for treating previously untreated patients with circulating tumor DNA (ctDNA) at high risk, the method comprising administering glimepiride in combination with rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP), wherein the patient is identified as having ctDNA if the ctDNA decreases by less than 2.5 log after one or two R-CHOP cycles.High risk.

[0406] Bispecific antibody therapies involving T cell activation, such as glimepiride, have been associated with cytokine release syndrome (CRS). CRS is a potentially life-threatening complex of symptoms caused by the excessive release of cytokines by immune effectors or target cells during an excessive and sustained immune response. CRS can be triggered by a variety of factors, including infection with toxic pathogens, or by agents that activate or enhance the immune response to produce a significant and sustained immune response.

[0407] Regardless of the triggering factor, severe or life-threatening CRS is a medical emergency. If not successfully managed, it can lead to significant disability or fatal outcomes. Current clinical management focuses on treating individual signs and symptoms, providing supportive care, and attempting to suppress the inflammatory response with high-dose corticosteroids. However, this approach is not always successful, especially in the case of late intervention. Furthermore, steroids may negatively affect T cell function, which may diminish the clinical benefits of immunomodulatory therapies in cancer treatment.

[0408] A. CRS Symptoms and Grading

[0409] CRS is graded according to the modified cytokine release syndrome grading system established by Lee et al., Blood, 124: 188-195, 2014 or Lee et al., Biol Blood Marrow Transplant, 25(4): 625-638, 2019, as described in Table 2. In addition to the diagnostic criteria, recommended CRS management based on severity (including early intervention with corticosteroids and / or anti-cytokine therapy) is also provided and mentioned in Tables 2 and 3.

[0410] Table 2. Description of the cytokine release syndrome grading system 36 / 71 pages 40 CN 122070142 A

[0411]

[0412] Lee 2014 criteria: Lee et al., Blood, 124: 188-195, 2014.

[0413] ASTCT consensus grading: Lee et al., Biol Blood Marrow Transplant, 25(4): 625-638, 2019.

[0414] a. Low-dose vasopressors: Single vasopressors at doses lower than those shown in Table 4.

[0415] b. High-dose vasopressors: As defined in Table 4.

[0416] Fever was defined as a body temperature ≥ 38°C that was not attributable to any other cause. In CRS patients who subsequently received antipyretics or anticytokine therapy (such as tocilizumab or steroids), fever was no longer required for grading the severity of subsequent CRS. In this case, CRS grading was driven by hypotension and / or hypoxia.

[0417] CRSThe level is determined by a more serious event: hypotension or hypoxia that cannot be attributed to any other cause. For example, CN 122070142 A, page 37 / 71 of the instruction manual, classifies a patient with a body temperature of 39.5°C, hypotension requiring one vasopressor, and hypoxia requiring a low-flow nasal cannula as a grade 3 CRS.

[0418] A low-flow nasal cannula is defined as delivering oxygen at a rate of ≤ 6 L / min. Low flow also includes blowing oxygen delivery, sometimes used in pediatrics. A high-flow nasal cannula is defined as delivering oxygen at a rate of > 6 L / min.

[0419] Table 3. High-dose vasopressors

[0420]

[0421] min = minutes; VASST = vasopressin and septic shock test.

[0422] A VASST vasopressor equivalence equation: Norepinephrine equivalent dose = [Norepinephrine (µg / min)] + [Dopamine (µg / kg / min) ÷ 2] + [Epinephrine (µg / min)] + [Phenylephrine (µg / min) ÷ 10].

[0423] Mild to moderate CRS and / or infusion-related reactions (IRR) may include symptoms such as fever, headache, and myalgia, and can be treated symptomatically with analgesics, antipyretics, and antihistamines as needed. Severe or life-threatening CRS and / or IRR manifestations, such as hypotension, tachycardia, dyspnea, or chest discomfort, should be aggressively treated with supportive and resuscitation measures as directed, including the use of high-dose corticosteroids, IV fluids, admission to the intensive care unit, and other supportive measures. Severe CRS may be associated with other clinical sequelae, such as disseminated intravascular coagulation, capillary leak syndrome, or macrophage activation syndrome (MAS). Standards of care for severe or life-threatening CRS resulting from immunotherapy have not been established; case reports and recommendations regarding the use of anticytokine therapies (such as tocilizumab) have been published (Teachey et al., Blood, 121: 5154–5157, 2013; Lee et al., Blood, 124: 188–195, 2014; Maude et al., New Engl J Med, 371: 1507–1517, 2014).

[0424] B. Pretreatment or management of CRS-related symptoms with tocilizumab

[0425] CRS is associated with high IL-6 levels (Panelli et al., J Transl Med, 2: 17, 2004; Lee et al., Blood, 124: 188-195, 2014; Doessegger and Banholzer, Clin Transl Immunology, 4: e39,2015), and IL-6 is associated with the severity of CRS, with patients experiencing severe or life-threatening CRS (NCI CTCAE grade 4 or 5) having much higher IL-6 levels compared to patients who did not experience CRS or experienced a milder CRS response (NCI CTCAE grade 0-3) (Chen et al., J Immunol Methods, 434: 1-8, 2016).

[0426] Tocilizumab (ACTEMRA® / ROACTEMRA®) is a recombinant humanized anti-human monoclonal antibody against soluble and membrane-bound IL-6R that inhibits IL-6-mediated signaling (see, for example, WO 1992 / 019579, which is incorporated herein by reference in its entirety). Tocilizumab has been approved by the U.S. Food and Drug Administration (FDA) for the treatment of adults and 2 Severe or life-threatening CAR-T cell-induced CRS in pediatric patients aged 6 years and older. Initial clinical data (Locke et al., Blood, 130: 1547, 2017) suggest that tocilizumab prophylaxis can reduce the severity of CAR-T cell-induced CRS by blocking IL-6 receptor signaling before cytokine release. Therefore, tocilizumab prophylaxis can also reduce the frequency or severity of CRS associated with bispecific antibody therapy. Other anti-IL-6R antibodies that can be used in combination with tocilizumab include sarilumab, vobarilizumab (ALX-0061), SA-237, and their variants.

[0427] In some aspects, an effective amount of tocilizumab is administered as a prophylaxis, for example, before administration of glimepiride to the subject. Administration of tocilizumab as a prophylaxis can reduce CRS. The frequency or severity of the occurrence. In some respects, tocilizumab is administered as a prodrug in cycle 1, such as before the first dose (C1D1), second dose (C1D2), and / or third dose (C1D3) of glimepiride. In some respects, tocilizumab is administered intravenously to the subject in a single dose of about 1 mg / kg to about 15 mg / kg (e.g., about 4 mg / kg to about 10 mg / kg, e.g., about 6 mg / kg to about 10 mg / kg, e.g., about 8 mg / kg). In some respects, tocilizumab is administered intravenously in a single dose of about 8 mg / kg.Tocilizumab is administered intravenously to the subject in a single dose form. Other anti-IL-6R antibodies that can be used in combination with tocilizumab include sarilumab, vobarilizumab (ALX-0061), SA-237, and variants thereof.

[0428] For example, in one embodiment, glimetuzumab is co-administered with tocilizumab (ACTEMRA® / ROACTEMRA®), wherein tocilizumab (ACTEMRA® / ROACTEMRA®) is first administered to the subject, followed by administration of the bispecific antibody alone (e.g., the subject is pre-treated with tocilizumab (ACTEMRA® / ROACTEMRA®)).

[0429] In another aspect, in subjects treated with glimetuzumab, tocilizumab is administered to treat or alleviate symptoms associated with CRS. If a subject has a grade 2 or higher CRS event with extensive comorbidity following administration of glimetuzumab, the method may further include administering a first dose of an IL-6R antagonist (e.g., an anti-IL-6R antibody, such as tocilizumab (ACTEMRA® / ROACTEMRA®)) to manage the grade 2 or higher CRS event while discontinuing treatment with glimetuzumab. In some cases, the first dose of tocilizumab is administered intravenously to the subject at a dose of approximately 8 mg / kg. Other anti-IL-6R antibodies that can be used in combination with tocilizumab include sarilumab, vobarilizumab (ALX-0061), SA-237, and variants thereof. In some cases, if a grade 2 or higher CRS event resolves to ≤ grade 1 CRS event within two weeks, the method may further include resuming treatment with glimetuzumab at a reduced dose. In some cases, if the event occurs during or within 24 hours of the infusion, a smaller dose is 50% of the initial infusion rate of the previous cycle. On the other hand, if a grade 2 or higher CRS event does not resolve or worsens to a grade ≥ 3 CRS event within 24 hours of treatment for the grade 2 or higher CRS event, the method may further include administering one or more (e.g., one, two, three, four, or five or more) additional doses of an IL-6R antagonist (e.g., an anti-IL-6R antibody, such as tocilizumab) to the subject to manage the grade 2 or ≥ 3 CRS event. In certain specific cases, a grade 2 or higher CRS event does not resolve or worsens to a grade ≥ 3 CRS event within 24 hours of treatment for the grade 2 or higher CRS event.Grade 2 or ≥ 3 CRS events, and the method may further include administering one or more additional doses of tocilizumab to the subject to manage Grade 2 or ≥ 3 CRS events. In some cases, one or more additional doses of tocilizumab are administered intravenously to the subject at a dose of about 1 mg / kg to about 15 mg / kg (e.g., about 4 mg / kg to about 10 mg / kg, e.g., about 6 mg / kg to about 10 mg / kg, e.g., about 8 mg / kg).

[0430] C. Other pretreatment instructions for CRS risk mitigation 39 / 71 pages 43 CN 122070142 A

[0431] In one embodiment, the treatment regimen provided herein further includes administering a prophylactic medication prior to administration of glimepiride. In one embodiment, the prophylactic medication includes corticosteroids (such as, e.g., prednisolone, dexamethasone, or methylprednisolone), acetaminophen / paracetamol, and / or antihistamines (such as, e.g., diphenhydramine). In one embodiment, a prophylactic medication is administered at least 60 minutes prior to administration of glimetuzumab (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 36, 48 hours or longer). In one embodiment, the treatment regimen further includes administering a prophylactic medication prior to administration of glimetuzumab. In embodiments, the prophylactic medication includes corticosteroids (such as, for example, prednisolone, dexamethasone, or methylprednisolone), antipyretics (such as, for example, acetaminophen / paracetamol), and / or antihistamines (such as, for example, diphenhydramine). In one embodiment, the subject receives a corticosteroid prophylactic medication prior to glimetuzumab. It has been demonstrated that, relative to methylprednisolone, the use of a dexamethasone prophylactic medication reduces the levels of glimetuzumab-induced cytokines in mice pretreated with dexamethasone. Therefore, in one embodiment, the corticosteroid is dexamethasone. In one embodiment, a prophylactic medication is administered at least 60 minutes prior to administration of glimepiride (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 36, 48 hours or longer). In another embodiment, a prophylactic medication is administered at least 60 minutes prior to each administration of glimepiride (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 36, 48 hours or longer). In yet another embodiment, a prophylactic medication in the form of a corticosteroid is administered before the first dose (C1D1) of the first cycle and the second dose (C1D2) of the first cycle, and before the first dose (C2D1) of the second cycle and the first dose (C3D1) of the third cycle, provided that no CRS has occurred in the previous cycle at the target dose.For patients who have reached and tolerated two subsequent cycles of doses, the prophylactic medication may be optional.

[0432] In one embodiment, the prophylactic medication is administered at least 60 minutes (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 36, 48 hours or longer) prior to the administration of pretreatment with an anti-CD20 antibody (particularly olibutuzumab).

[0433] In one embodiment, a corticosteroid is administered to control any associated adverse events following administration of the anti-CD20 / anti-CD3 bispecific antibody (e.g., glimetuzumab).

[0434] (v) Administration of the anti-CD20 / anti-CD3 bispecific antibody

[0435] The method may involve administering glimetuzumab (and / or any additional therapeutic agent) by any suitable means, including parenteral, intrapulmonary and intranasal administration, and, if local treatment is required, intralesional administration. Parenteral infusion includes intravenous, subcutaneous, intramuscular, intra-arterial, and intraperitoneal administration routes. In some embodiments, glimetuzumab is administered via intravenous infusion.

[0436] In one embodiment, the infusion time of the anti-CD20 / anti-CD3 bispecific antibody (particularly glimetuzumab) is at least 4 hours (e.g., about 4 hours, about 4.5 hours, about 5 hours, about 5.5 hours, or about 6 hours). In a particular embodiment, the infusion duration of glimetuzumab is about 4 hours. In one embodiment, the infusion time of glimetuzumab may be shortened or prolonged. In one embodiment (e.g., in the absence of infusion-related adverse events), the infusion time of glimetuzumab in subsequent cycles is reduced to 2 hours ± 15 minutes. In one embodiment, the infusion time is increased to a maximum of 8 hours (e.g., about 4 hours, about 5 hours, about 6 hours, about 7 hours, or about 8 hours) (e.g., for high-risk subjects with a history of CRS). In one embodiment, for example, for patients at increased risk of CRS, patients who have experienced IRR or CRS with their previous dose of glimetuzumab, or patients at increased risk of recurrence of IRR / CRS at subsequent doses, the infusion time of glimetuzumab is extended to a maximum of 8 hours.

[0437] For all methods described herein, glimetuzumab is formulated, administered, and given in accordance with good medical practice. Factors to be considered in this context include the specific condition being treated, the specific mammal being treated, the individual subject's clinical condition, the cause of the condition, the site of delivery of the drug, the method of administration, the administration regimen, and any other instructions known to the physician. 40 / 71 pages 44 CN 122070142 AFactors. Glimetabolumab is not mandatory but optional to be co-formulated with one or more agents currently used for the prevention or treatment of the disease in question. The effective amount of such other agents depends on the amount of glimetabolumab present in the formulation, the type of disease or treatment, and other factors discussed above. Glimetabolumab can be appropriately administered to the subject through a series of treatments.

[0438] Another aspect of the invention relates to the invention as described above.

[0439] Examples

[0440] Some embodiments of the technology described herein can be defined according to any of the following numbered examples:

[0441] 1. A method of treating a previously untreated patient with high-risk circulating tumor DNA (ctDNA) diffuse large B-cell lymphoma, the method comprising administering a combination of a bispecific antibody targeting CD3 and CD20 containing the VH and VL sequences of glimetabolumab with chemotherapy, wherein the patient has been identified as high-risk for ctDNA if the ctDNA decreases by less than 2.5-log after one or two cycles of chemotherapy.

[0442] 2. The method according to Example 1, wherein the bispecific antibody targeting CD3 and CD20 comprises the heavy chain of SEQ ID NO: 17, the heavy chain of SEQ ID NO: 18, the two light chains of SEQ ID NO: 20 and the light chain of SEQ ID NO: 19; and wherein the heavy chain and the light chain are assembled to form a first Fab molecule specifically binding to CD20, a second Fab molecule specifically binding to CD3 and a third Fab molecule specifically binding to CD20, and

[0443] an Fc domain composed of a first subunit and a second subunit capable of stable association.

[0444] 3. The method according to Example 1 or 2, wherein the chemotherapy is selected from: rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP); rituximab, cyclophosphamide, doxorubicin and prednisone (R-CHP); or rituximab, ifosfamide, carboplatin and etoposide phosphate (R-ICE)

[0445] 4. The method according to any one of Examples 1 to 3, wherein the bispecific antibody targeting CD3 and CD20 is glimepiride, and the chemotherapy is rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP), and wherein the patient has been identified as high-risk for ctDNA if the ctDNA decreases by less than 2.5 log after one or two R-CHOP cycles.

[0446] 5. The method according to Example 4, wherein by measuring before or on the day of the first treatment (baseline) and at least oneThe amount of ctDNA after an R-CHOP treatment cycle is used to determine the ctDNA reduction.

[0447] 6. The method according to Example 4 or 5, wherein the amount of ctDNA is determined on or before day 1 of the first R-CHOP treatment cycle (baseline) and on day 1 of the second R-CHOP treatment cycle.

[0448] 7. The method according to any one of Examples 4 to 6, wherein if the patient's ctDNA reduction is less than 2 log, the patient is identified as high-risk for ctDNA.

[0449] 8. A method for treating previously untreated diffuse large B-cell lymphoma, the method comprising:

[0450] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0451] b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0452] c. selecting patients whose ctDNA decreases by <2.5 log between baseline and after at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimepiride.

[0453] 9. The method according to Example 8, wherein step a) comprises administering R-CHOP on day 1 of cycles 1, 2, and optionally cycle 3. Instructions for Use, Pages 41 / 71, CN 122070142 A

[0454] 10. The method according to Example 9, wherein step a) includes administering R-CHOP on day 1 of cycles 1 and 2.

[0455] 11. The method according to any one of Examples 8 to 10, wherein step b) includes determining the amount of ctDNA on day 1 or before (baseline) of cycle 1 of the R-CHOP treatment in step a) and on day 1 of cycle 2 or day 1 of cycle 3.

[0456] 12. The method according to Example 11, wherein step b) includes determining the amount of ctDNA on day 1 (baseline) of cycle 1 of the R-CHOP treatment in step a) and on day 1 of cycle 2.

[0457] 13. The method according to any one of Examples 8 to 12, wherein step c) includes selecting patients with a ctDNA reduction of <2-log.

[0458] 14. The method according to any one of Examples 8 to 13, wherein step c) comprises continuing treatment with R-CHOP for a total of six cycles (cycles 1 to 6), and adding glimepiride for a total of8 cycles (cycles 3 to 10).

[0459] 15. The method according to any one of Examples 8 to 14, wherein glimepiride is administered at a dose of 2.5 mg on day 8 of cycle 3 and at a dose of 10 mg on day 15, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 30 mg on day 1 of cycles 7 to 10.

[0460] 16. The method according to any one of the preceding examples, wherein the ctDNA is circulating tumor DNA in plasma.

[0461] 17. The method according to any one of the preceding examples, wherein the amount of ctDNA is measured in a blood sample from the patient.

[0462] 18. The method according to Example 17, wherein the amount of ctDNA is measured in genomic DNA isolated from peripheral mononuclear cells or plasma-depleted whole blood (PDWB) or in cell-free DNA (cfDNA) in plasma.

[0463] 19. The method according to Example 18, wherein the ctDNA is measured in cell-free plasma DNA.

[0464] 20. The method according to any of the preceding examples, wherein the ctDNA is determined by targeted next-generation sequencing (NGS).

[0465] 21. The method according to any of Examples 4 to 20, wherein rituximab is administered intravenously at a dose of 375 mg / m2, cyclophosphamide is administered intravenously at a dose of 750 mg / m2, doxorubicin is administered intravenously at a dose of 50 mg / m2, vincristine is administered via IV bolus at a dose of 1.4 mg / m2, and prednisone is administered at a dose of 100 mg / day on days 1 to 5.

[0466] 22. The method according to any of Examples 1 to 21, wherein the length of each cycle is 21 days.

[0467] 23. The method according to any one of Examples 4 to 22, wherein administration of such treatment to multiple human patients at high risk of ctDNA produces an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%.

[0468] 24. The method according to Example 23, wherein the ORR is at least 95%.

[0469] 25. The method according to any one of Examples 1 to 21, wherein administration of such treatment to multiple human patients at high risk of ctDNA produces a complete response (CR) rate of at least 75%, at least 80%, or at least 85%.

[0470] 26. The method according to Example 25, wherein the CR rate is at least 85%.

[0471] 27. The method according to Examples 23 and 24 or Examples 25 and 26, wherein the ORR or CR rate is determined at the end of treatment.

[0472] 28. The method according to Example 27, wherein the ORR or CR is determined by PET-CT scan.

[0473] 29. The method according to Example 27 or 28, wherein the ORR or CR is determined according to the Lugano response criteria as specified in the 2014 specification, page 42 / 71, 46 CN 122070142 A.

[0474] 30. Glimetuzumab, in a method of treating previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, the method comprising administering a combination of glimetuzumab and chemotherapy to a patient, wherein the patient has been identified as at high risk of ctDNA if the ctDNA decreases by less than 2.5 log after one cycle of chemotherapy.

[0475] 31. Glimetrozine, in a method of treating previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, the method comprising:

[0476] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0477] b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0478] c. selecting patients with a ctDNA reduction of <2.5 log between baseline and at least one R-CHOP treatment cycle and continuing treatment with R-CHOP and glimetrozine.

[0479] 32. Use of glimetuzumab in the manufacture of a medicament for treating a subject with previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, wherein the patient is administered a combination of glimetuzumab and chemotherapy, wherein the patient has been identified as at high risk of ctDNA if the ctDNA decreases by less than 2.5 log after one cycle of chemotherapy.

[0480] 33. Use of glimetuzumab in the manufacture of a medicament for treating a subject with previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, comprising:

[0481] a. administering rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles;

[0482] b. determining the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and

[0483] c. selecting at baseline and at least one R-CHOP treatment cycle.Patients with a ctDNA reduction of <2.5-log between treatment cycles continued to be treated with R-CHOP and glimetuzumab.

[0484] 34. The present invention as described above.

[0485] Examples

[0486] The following are examples of the methods and compositions of the present invention. It should be understood that various other examples may be implemented in view of the general description provided above.

[0487] Example 1. A Phase II study evaluating the safety and efficacy of glimetuzumab in combination with rituximab (R) plus cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP) in patients with untreated diffuse large B-cell lymphoma at high risk of circulating tumor (ct) DNA

[0488] Reasons for the study

[0489] The purpose of this study was to evaluate the safety, efficacy and pharmacokinetics of the combination of glimetuzumab and R-CHOP in patients at high risk of ctDNA. Patients at high risk for ctDNA are those who fail to achieve a ≥2 log-fold reduction in ctDNA levels between Day 1 of Cycle 1 (pre-treatment) and Day 1 of Cycle 2. ctDNA is a novel biomarker that offers the possibility of early and sensitive identification of patients with DLBCL at high risk of first-line treatment failure. In the first-line setting, a subset of patients with DLBCL has a poor prognosis after treatment with R-CHOP immunochemotherapy. Nevertheless, R-CHOP remains the state-of-the-art (SOC) treatment for DLBCL. To date, efforts to define baseline prognostic scores and develop biomarkers to improve the International Prognostic Index (IPI) and guide treatment decisions have been unsuccessful. Compared to current baseline prognostic factors, ctDNA has a significant advantage in allowing dynamic assessment early in the patient’s treatment process and in doing so, identifying newly diagnosed individuals who are poorly responsive to immunochemotherapy, who may progress without changing their treatment strategy, and therefore may benefit from therapy intensive with glimetuzumab.

[0490] Objectives

[0491] This Phase II, open-label, multicenter study evaluated the safety, efficacy, and pharmacokinetics of the combination of glimetuzumab and R-CHOP in a first-line setting in individuals with high-risk DLBCL and ctDNA. The specific objectives and corresponding endpoints of the study are outlined in the table below (Table 4). In this protocol, “study treatment” refers to the combination of treatments (i.e., R-CHOP, with or without glimetuzumab) assigned to participants as part of this study.

[0492] Table 4. Objectives and Endpoints 44 / 71 pages 48 CN 122070142 A

[0493] Instructions for Use, Pages 45 / 71, CN 122070142 A

[0494] ADA = Anti-drug antibody; aLugano = Automated response according to the 2014 Lugano Response Criteria; ASTCT = American Society for Transplantation and Cell Therapy; aTMTV = Automated Total Metabolic Tumor Volume; AUC = Area Under the Concentration-Time Curve; Cmax = Maximum Concentration; CR = Complete Response; CRS = Cytokine Release Syndrome; CT = Computed Tomography; CTCAE = Common Terminology Criteria for Adverse Events; ctDNA = Circulating Tumor DNA; DLBCL = Diffuse Large B-Cell Lymphoma; DOR = Duration of Response; EOT = End of Treatment; NCI = National Cancer Institute; ORR = Objective Response Rate; OS = Overall Survival; PET = Positron Emission Tomography; PFS = Progression-Free Survival; PK = Pharmacokinetics; R-CHOP = Rituximab in combination with cyclophosphamide, doxorubicin, vincristine, and prednisone; SUVmax = maximum standardized uptake value; TMTV = total metabolic tumor volume.

[0495] Overall Study Design

[0496] This was a phase II, multicenter study evaluating the safety and efficacy of glimetuzumab in combination with R-CHOP immunochemotherapy in individuals with untreated DLBCL who were assessed as high-risk based on ctDNA. Patients were classified as high-risk for R-CHOP treatment failure if they showed a <2-log (100 × 100) decrease in ctDNA levels between baseline (before day 1 of cycle 1) and after one cycle of R-CHOP treatment (before day 1 of cycle 2) using the experimental AOA-NHL test.

[0497] Patients at high-risk for ctDNA were enrolled before day 8 of cycle 3 and received glimetuzumab at escalating doses (2.5 mg, 10 mg, and 30 mg) starting on day 8 of cycle 3. For the purposes of this study, individuals whose ctDNA levels decreased by at least 2-log-fold between baseline (before Day 1 of Cycle 1) and after one cycle of R-CHOP treatment (before Day 1 of Cycle 2) were considered to be at lower risk of R-CHOP treatment failure and were ineligible for enrollment or rescreening. For enrollment in Cycle 3, eligible patients were treated with standard of care (SOC) doses of R-CHOP starting in Cycle 1. During Cycles 3 through 6, participants received combination therapy with R-CHOP and glimepiride, and during Cycles 7 through 10...Participants received glimetuzumab monotherapy as consolidation during the study. PET / CT scans were performed at the end of treatment (EOT), and early-mid-term PET / CT response assessments were conducted after cycle 2; results were used to evaluate the correlation between imaging findings and ctDNA levels. To reduce the risk of CRS, glimetuzumab was administered using an escalating dosing regimen consisting of 2.5 mg and 10 mg doses administered on days 8 and 15 of cycle 3, respectively. The total dose administered in cycle 3 did indeed exceed the dose for which safety was established in the NP40126 study. For subsequent cycles, 30 mg of glimetuzumab was administered on day 8 of cycles 5 and 6, and on day 1 of cycles 7 through 10. All participants were closely monitored for adverse events throughout the study and for at least 90 days after the last dose of study treatment. Adverse events were classified according to the National Cancer Institute Adverse Events General Toxicity Criteria version 5.0 (NCI CTCAE v5.0), page 46 / 71, CN 122070142 A, and CRS was classified according to the American Society for Transplantation and Cell Therapy (ASTCT) 2019 CRS Consensus Classification. An Internal Monitoring Committee (IMC) was established to monitor patient safety during the study and to make recommendations on study practices based on trial safety data to ensure patient safety during the study treatment.

[0498] Participants’ radiation-induced tumor response was assessed by PET / CT and CT scans at the end of Cycle 2 (ctDNA screening), after treatment, and during the study follow-up period. Tumor response was assessed according to the investigator’s 2014 Lugano Response Criteria for malignant lymphoma (hereinafter referred to as the 2014 Lugano Response Criteria).

[0499] The study protocol is provided in Figure 2, and the dosing regimen is shown in Figure 3.

[0500] Study Treatment

[0501] The study drugs (IMPs) in this study were glimetuzumab and tocilizumab. R-CHOP and any pre-treatment with corticosteroids were considered non-study drugs. CNS prophylaxis using intrathecal methotrexate, neutropenia prophylaxis using granulocyte colony-stimulating factors (G-CSFs), and dexamethasone were also considered non-study drugs.

[0502] Gglimetuzumab

[0503] Gglimetuzumab is a human IgG1 bispecific antibody that targets CD20 expressed on the surface of B cells and CD3ε expressed on the surface of T cells. Gglimetuzumab induces tumor cell lysis by binding to CD20 on tumor cells, and simultaneously achieves T cell activation, proliferation, and cell regeneration by binding to CD3ε on the T cell receptor complex.Factor release. Griffithomab has a unique structure containing two CD20-binding domains and one CD3-binding domain (2:1 design), which significantly enhances its affinity for CD20 compared to the 1:1 form. This novel antibody has demonstrated a manageable safety profile and is currently under rigorous investigation for its use in NHL as a single agent and as a combination partner with other therapies.

[0504] Griffithomab is a bispecific antibody comprising two Fab molecules that specifically bind to CD20, each Fab molecule containing the following six hypervariable regions (HVRs):

[0505] HVR-H1: YSWIN (SEQ ID NO: 1); HVR-H2: RIFPGDGDTDYNGKFKG (SEQ ID NO: 2); HVR-H3: NVFDGYWLVY (SEQ ID NO: 3); HVR-L1: RSSKSLLHSNGITYLY (SEQ ID NO: 4); HVR-L2: QMSNLVS (SEQ ID NO: 5); and HVR-L3: AQNLELPYT (SEQ ID NO: 6)

[0506] Griffithomab further comprises at least one Fab molecule that specifically binds to CD3, each Fab molecule containing the following six hypervariable regions (HVRs): HVR-H1: TYAMN (SEQ ID NO: 1); HVR-H2: TYAMN (SEQ ID NO: 1); HVR-H3: TYAMN (SEQ ID NO: 1); HVR-H4: TYAMN (SEQ ID NO: 1); HVR-H5: TYAMN (SEQ ID NO: 1); HVR-H6: TYAMN (SEQ ID NO: 1); HVR-H7: TYAMN (SEQ ID NO: 1); HVR-H8: TYAMN (SEQ ID NO: 1); HVR-H9: TYAMN (SEQ ID NO: 1); HVR-H1: TYAMN (SEQ ID NO: 1); HVR-H2: TYAMN (SEQ ID NO: 1); HVR-H3: TYAMN (SEQ ID NO: 1); HVR-H4: TYAMN (SEQ ID NO: 1); HVR-H5: TYAMN (SEQ ID NO: 1); HVR-H3: TYAMN (SEQ ID NO: 1); HVR-H4: TYAMN (SEQ ID 9); HVR-H2: RIRSKYNNYATYYADSVKG (SEQ ID NO: 10); HVR-H3: HGNFGNSYVSWFAY (SEQ ID NO: 11); HVR-L1: GSSTGAVTTSNYAN (SEQ ID NO: 12); HVR-L2: GTNKRAP (SEQ ID NO: 13); and HVR-L3: ALWYSNLWV (SEQ ID NO: 14).

[0507] Griffithromab was administered intravenously in an escalating dosing regimen, starting with 2.5 mg on day 8 of cycle 3 and 10 mg on day 15 of cycle 3, followed by 30 mg on day 8 of cycles 4 to 6 and day 1 of cycles 7 to 10, with each cycle lasting 21 days (i.e., every 3 weeks). Additional escalating doses (e.g., on day 4 of cycle 3) or fractionated administration (e.g., 0.5 mg on day 8 of cycle 3 and 2 mg on day 9 of cycle 3) may also be introduced.

[0508] A 20 mg IV dexamethasone predrug is administered 1 hour before administration of glimetuzumab;Oral acetaminophen or paracetamol (500 mg or 1000 mg) and an antihistamine such as diphenhydramine (50 mg) were administered approximately 30 minutes before the start of infusion.

[0509] Initially, glimetuzumab was administered to participants over 4 hours (±15 minutes) on days 8 and 15 of cycle 3, days 8 of cycles 4 to 6, and day 1 of cycles 7 to 10. For participants with increased CRS risk, or those who experienced CRS at previous doses of glimetuzumab or, in the investigator's judgment, had an increased risk of recurrent CRS at subsequent doses, the infusion time was extended to a maximum of 8 hours. In the absence of infusion-related adverse events or CRS, the infusion time of glimetuzumab in cycles 5 and 6 may be reduced to 2 hours (±15 minutes) in the investigator's judgment. For cycles 7, 8, 9, and 10, in the absence of infusion-related adverse events or CRS, the infusion time of glimetuzumab may be reduced to 90 minutes (±30 minutes) at the investigator's discretion.

[0510] For participants who tolerate two 30 mg doses of glimetuzumab without CRS, pre-treatment with dexamethasone during subsequent cycles is optional, at the investigator's discretion. However, if a subject experiences CRS, pre-treatment with dexamethasone is necessary for subsequent doses of glimetuzumab until no additional CRS event is observed.

[0511] Tocilizumab

[0512] Tocilizumab is a recombinant humanized anti-human monoclonal antibody that targets the soluble and membrane-bound IL-6 receptor, inhibiting IL-6-mediated signaling. If necessary, tocilizumab may be administered to participants who experience CRS events as a rescue investigational drug (IMP). Tocilizumab was administered at a dose of 8 mg / kg to participants ≥ 30 kg and at a dose of 12 mg / kg to participants < 30 kg. A dose exceeding 800 mg per infusion is not recommended.

[0513] Rituximab

[0514] Rituximab was administered intravenously to participants every 21 days at a dose of 375 mg / m² on day 1 of cycles 1 through 6, together with CHOP.

[0515] Rituximab preparation and administration were performed according to local prescribing information, including prophylactic medication. Subcutaneous administration of rituximab was not permitted in this study. Locally approved standard-of-care biosimilars of rituximab were permitted.

[0516] CHOP Chemotherapy

[0517] CHOP chemotherapy consisted of IV cyclophosphamide, IV doxorubicin, and IV...The treatment consists of vincristine administered via bolus and prednisone or prednisolone administered orally. The standard CHOP is administered as follows for six 21-day cycles (including cycles 1 and 2 during ctDNA screening):

[0518] Cyclophosphamide: 750 mg / m² intravenously on day 1

[0519] Doxorubicin: 50 mg / m² intravenously on day 1 or according to institutional guidelines

[0520] Vincristine: 1.4 mg / m² via IV bolus on day 1, with a recommended upper limit of 2 mg.

[0521] Prednisone: 100 mg / day orally from day 1 to day 5 (prednisone on day 1 can be administered intravenously, and the remaining dose on days 2 to 5 can be administered orally).

[0522] In countries where prednisone is unavailable or not the preferred treatment, prednisone is replaced with prednisolone (1:1 conversion; 100 mg); in countries or regions where prednisone is unavailable, 80 mg of methylprednisolone is used instead.

[0523] The maximum permissible dose of vincristine is 2 mg, based on investigator judgment. For participants ≥ 70 years of age, the maximum permissible dose of vincristine is 1.5 mg. The maximum permissible body surface area is 2 m², based on institutional standards.

[0524] When rituximab is administered on the same day as the CHOP program, it is recommended to administer prednisone (100 mg) before rituximab infusion.

[0525] When rituximab is administered on the same day as the CHOP program, it is recommended to administer prednisone (100 mg) before rituximab infusion. If the rituximab dose on day 1 of cycle 1 is split, both infusions must be performed with appropriate prophylaxis and in accordance with the infusion guidelines described in the pharmacy manual or product characteristics summary. If the CHOP start date is later than day 1 of cycle 1, the planned day 1 of the next cycle 1 is calculated from the start date of the CHOP, see Instructions for Use, page 48 / 71, 52 CN 122070142 A, to maintain the usual 21-day chemotherapy interval.

[0526] Intrathecal Methotrexate

[0527] For patients with high risk of CNS IPI (defined as having four or more of the following risk factors: age > 60 years, stage III or IV disease, more than two extranodal sites, longest diameter greater than the upper limit of normal, or kidney or adrenal involvement), intrathecal methotrexate is recommended (but not mandatory) for CNS prophylaxis and may be included as part of the patient’s protocol at the investigator’s discretion. If CNS prophylaxis is administered, the dosage and protocol should be clearly documented and administered in accordance with local guidelines.

[0528] Granulocyte colony-stimulating factor

[0529] To minimize the incidence, severity, and duration of neutropenia-related complications, all participants received primary prophylaxis with G-CSF during cycles 1 through 6.

[0530] G-CSF must be used during cycles 3 through 6, i.e., any of the following:

[0531] Filgrastim: 5 μg / kg (rounded to 300 μg or 480 μg) subcutaneously once daily for up to 48 hours after R-CHOP administration (day 1 of cycles 1 through 6) for 7 days or until ANC > 1.5 × 10⁹ / L for 2 consecutive days

[0532] Glycolized filgrastim: 6 mg subcutaneously after R-CHOP administration (day 1 of cycles 1 through 6) for up to 48 hours

[0533] Leglucopyridine is an acceptable alternative.

[0534] G-CSF may be administered intravenously if preferred or clinically indicated.

[0535] Dexamethasone

[0536] Dexamethasone is administered 1 hour prior to administration of glibenclamide; oral acetaminophen or paracetamol (500 mg or 1000 mg) and antihistamines such as diphenhydramine (50 mg) are administered approximately 30 minutes prior to the start of infusion.

[0537] AVENIO Oncology Assay for Non-Hodgkin Lymphoma

[0538] The AVENIO Oncology Assay for Non-Hodgkin Lymphoma (AOA-NHL) uses targeted next-generation sequencing (NGS) and sequences genomic DNA and cell-free plasma DNA (cfDNA) isolated from peripheral mononuclear cells or plasma-depleted whole blood (PDWB) to qualitatively detect ctDNA as an indicator of minimal residual disease (MRD). The AOA-NHL test can be used to determine the ctDNA status of patients with DLBCL.

[0539] The AOA-NHL test is an NGS tumor profiling and monitoring assay that identifies somatic DNA mutations in individuals with DLBCL by sequencing cfDNA isolated from plasma samples from patients at different time points during treatment. cfDNA consists of DNA fragments released into the bloodstream due to cell death in both normal and tumor tissues. The presence of somatic variants (non-individually inherited variants) in the tumor-derived DNA is assumed and referred to as ctDNA.

[0540] The AOA-NHL test is a laboratory-developed assay available at Roche Molecular Systems (San Jose, California, USA).The AOA-NHL test provides a simplified end-to-end NGS solution from DNA extraction to bioinformatics data analysis and reporting. The test is based on iDES-enhanced CAPP-Seq (Integrated Digital Error Suppression Enhanced Cancer Personalized Deep Sequencing Analysis) technology, described in detail by Newman et al. (2014, 2016). Multiple studies have demonstrated that ctDNA deep sequencing can detect somatic variants with very low allele fractions (Schwarzenbach et al. 2011; Narayan et al. 2012; Diehn et al. 2017). This makes ctDNA assays suitable for monitoring the disease burden of patients receiving treatment (Chin et al. 2019). Specification 49 / 71 pages 53 CN 122070142 A

[0541] The test is designed to report the presence of tumor-derived DNA variants spanning approximately 314 kilobases across 466 genomes associated with disease presence and to assess changes in the levels of such variants in the patient’s peripheral blood throughout treatment. The AOA-NHL test uses a proprietary bioinformatics workflow to identify tumor-specific variants (TSVs) by subtracting the variants present in a matching normal PDWB sample from those variants detected in plasma samples. The workflow also monitors for the same TSVs from plasma samples collected at subsequent time points. The mean allele fraction of all detected TSVs is calculated to estimate the ctDNA level per milliliter of plasma. The detection of these TSVs or a subset of these TSVs in a plasma sample indicates the presence of ctDNA in the patient's blood. Statistical significance of TSVs is expressed as a p-value, which is calculated using a Monte Carlo statistical method (Newman et al. 2016) to account for background variance.

[0542] The use of the AOA-NHL test is experimental and is intended to identify patients with untreated DLBCL at high-risk ctDNA levels who may be eligible to participate in this study. The test is limited to centralized analysis at Roche Molecular Systems in San Jose, California, a company accredited by the Clinical Laboratory Improvement Act of 1988 (CLIA) to perform highly complex clinical tests.

[0543] Study Duration

[0544] The total duration of participation for each individual is expected to be approximately 24 months.

[0545] Study Population

[0546] Approximately 40 participants with untreated DLBCL at high risk of ctDNA will be enrolled in this study.

[0547] Prospective approval of protocol deviations from recruitment and inclusion criteria is not permitted; this is referred to as a protocol waiver or exemption.

[0548] Inclusion Criteria

[0549] Participants were eligible for inclusion in the study only if they met all of the following criteria:

[0550] Participants aged ≥ 18 years

[0551] Life expectancy ≥ 6 months

[0552] Patients with previously untreated CD20-positive DLBCL, including those with one of the following diagnoses made according to the 2016 WHO classification of lymphomas (Swerdlow et al. 2016):

[0553] – Nonspecific DLBCL, including GCB and ABC / non-GCB types and dual-expressing lymphoma (MYC and BCL2 co-expression)

[0554] – High-grade B-cell lymphoma (HGBCL) with MYC and BCL2 and / or BCL6 translocations

[0555] The number of participants with HGBCL was capped at 15% of the screening population.

[0556] – Patients with newly diagnosed transformed follicular lymphoma

[0557] Eastern Cooperative Oncology Group (ECOG) performance status 0-2

[0558] IPI 1-5,

[0559] – Patients with IPI of 0 and with large lesions (> 6 cm) are eligible.

[0560] Submit sufficient biomarker blood samples for screening to determine ctDNA status before starting R-CHOP on Day 1 of Cycle 1 and on Day 1 of Cycle 2.

[0561] At least one two-dimensional FDG-avid measurable lymphoma lesion on PET / CT scan, defined as having a longest diameter > 1.5 cm on CT scan.

[0562] Left ventricular ejection fraction (LVEF) ≥ 50%, determined on cardiac multi-gated acquisition (MUGA) scan or cardiac echocardiography (ECHO).

[0563] Sufficient hematological function (unless there is evidence of extensive bone marrow infiltration or hypersplenism secondary to the underlying lymphoma, in which case blood products are permitted), defined as follows: Instructions for Use 50 / 71 pages 54 CN 122070142 A

[0564] – Hemoglobin ≥ 9.0 g / dL (≥ 90 g / L), and no infusion of concentrated RBCs in the 7 days prior to the first study treatment.

[0565] – ANC ≥ 1.0 × 10⁹ / L (≥ 1000 / µL)

[0566] – Platelet count ≥ 75 × 10⁹ / L (≥ 75,000 / µL)

[0567] Additional inclusion criteria for participants at high risk of ctDNA

[0568] After two R-CHOP cycles, participants are eligible to continue study enrollment only if they meet the following inclusion criteria:

[0569] Assess ctDNA using the experimental AOA-NHL testHigh-risk plasma samples were defined as those with a <2-log-fold decrease in ctDNA levels between Day 1 of Cycle 1 and Day 1 of Cycle 2, as assessed by a central laboratory.

[0570] Exclusion Criteria

[0571] Participants were excluded from the study if any of the following criteria were met:

[0572] Current diagnosis of: unclassifiable B-cell lymphoma (grey zone lymphoma) with intermediate features between DLBCL and classical Hodgkin lymphoma, primary mediastinal (thymic) large B-cell lymphoma, T-cell / histiocytic abundant large B-cell lymphoma, Burkitt lymphoma, CNS lymphoma (primary or secondary involvement), primary exudative DLBCL, and primary cutaneous DLBCL.

[0573] Contraindications to any of the individual components of R-CHOP, including prior anthracycline therapy.

[0574] History of severe allergy or anaphylactic reaction to mouse monoclonal antibodies, or known sensitivity or allergy to mouse products

[0575] Previous treatment for indolent lymphoma

[0576] Previous use of any monoclonal antibody within 4 weeks or 5 half-lives (whichever is shorter) prior to the start of Cycle 1, and / or prior use of any investigational therapy for cancer treatment within 28 days prior to the start of Cycle 1

[0577] Previous solid organ or allogeneic stem cell transplantation

[0578] Received a live vaccine (currently available SARS-CoV-2 vaccines, including mRNA vaccines and viral vector vaccines, are not live vaccines) within 28 days prior to the start of Cycle 1

[0579] Previous therapy for DLBCL and HGBCL, except for palliative short-term corticosteroid therapy

[0580] Patients requiring control of lymphoma symptoms during screening may receive steroids as follows:

[0581] Up to 1 mg / kg may be used during screening after a diagnostic biopsy. Prednisone or its equivalent (up to 100 mg / day) for 7 days to control lymphoma symptoms. For doses >1 mg / kg, consult a medical monitor.

[0582] Patients receiving ≤30 mg / day of prednisone or its equivalent corticosteroids for reasons other than controlling lymphoma symptoms (e.g., rheumatoid arthritis) must have documented evidence of a stable dose for at least 4 weeks prior to the start of Cycle 1.

[0583] Orthostatic hypotension and adrenal insufficiency may be managed with mineralocorticoids and corticosteroids, respectively.

[0584] Systemic immunosuppressants (including, but not limited to, cyclophosphamide, imidazoline, methotrexate, thalidomide, and anti-tumor necrosis factor agents) must be administered within 4 weeks prior to the first dose of R-CHOP.

[0585] Blood transfusions must be administered within 7 days prior to screening unless there is lymphoma involving the bone marrow or spleen (in which case blood transfusions are permitted).Evidence supporting the product)

[0586] Any of the following laboratory test values:

[0587] – Serum creatinine clearance <40 mL / min (calculated according to the Cockcroft-Gault formula) – AST or ALT > 2.5 × Upper Limit of Normal (ULN)

[0588] – Total bilirubin > 1.5 × ULN (participants with documented Gilbert's disease are enrolled if their total bilirubin ≤ 3 × ULN)

[0589] – INR or PT > 1.5 times ULN in the absence of therapeutic anticoagulation

[0590] – PTT or aPTT > 1.5 times ULN in the absence of therapeutic anticoagulation or lupus anticoagulation

[0591] Recent major surgery (within 6 days before the start of Day 1 of Cycle 1)

[0592] A history of serious or long-term cardiovascular disease, such as New York Heart Association Class III or IV heart disease or objective assessment Class C or D, myocardial infarction within the past 6 months, unstable arrhythmia, or unstable angina.

[0593] Other cancer history within 2 years prior to screening, excluding cervical carcinoma in situ, non-melanoma skin cancer, or stage I uterine cancer that has been appropriately treated.

[0594] Patients with malignancies treated only with radical surgery are ineligible unless there is a record of cancer remission for ≥2 years prior to enrollment without treatment.

[0595] Patients with current low-grade early prostate cancer (Gleason score ≤ 6, stage 1 or 2) that did not require treatment at any time prior to the study are eligible.

[0596] Any active infection that could affect patient safety within 7 days prior to Day 1 of Cycle 1 or at enrollment

[0597] – In the event of a pandemic or epidemic, screening for active infection should be considered according to local or institutional guidelines or guidelines of applicable professional societies (e.g., American Society of Clinical Oncology [ASCO] or European Society for Medical Oncology [ESMO]).

[0598] Positive Hepatitis B Surface Antigen (HBsAg) Test Results at Screening

[0599] – Patients with occult or prior hepatitis B virus (HBV) infection (defined as HBsAg negative and total hepatitis B core antibody [HBcAb] positive) may be included if they are willing to undergo monthly DNA testing. Patients with a protective titer of hepatitis B surface antibody (HBsAb) after vaccination or those with prior but cured hepatitis B are eligible.

[0600] Hepatitis C Virus (HCV) at ScreeningPositive antibody test result

[0601] – Patients with other positive HCV antibodies are only eligible for inclusion if the HCV RNA polymerase chain reaction (PCR) result is negative.

[0602] Suspected or latent tuberculosis

[0603] History of progressive multifocal leukoencephalopathy

[0604] Confirmed or suspected chronic active Estane-Barr virus infection

[0605] Known or suspected history of HLH

[0606] Clinically significant history of liver disease, including viral hepatitis or other hepatitis, or cirrhosis

[0607] Substance abuse within the 12 months prior to screening, including over-the-counter drugs and alcohol dependence

[0608] Any serious medical condition or abnormality in clinical laboratory tests that, in the investigator's judgment, would impair the patient's safe participation in and completion of the study, including severe psychosis requiring hospitalization and antipsychotic medication

[0609] Patients with a history of autoimmune diseases, including but not limited to: myasthenia gravis, myositis, autoimmune hepatitis, systemic lupus erythematosus, rheumatoid arthritis, inflammatory bowel disease, vascular thrombosis associated with antiphospholipid syndrome, Wegener's granulomatosis, Sjögren's syndrome, Guillain-Barré syndrome, multiple sclerosis, vasculitis, or glomerulonephritis, may be excluded from the following:

[0610] – Patients with a history of autoimmune-related hypothyroidism to a stable dose of thyroid replacement hormone (CBT) 52 / 71 pages 56 CN 122070142 A

[0611] – Patients with type 1 diabetes who are receiving an insulin regimen are eligible for this study.

[0612] Patients with a history of disease-associated immune thrombocytopenic purpura, autoimmune hemolytic anemia, or other stable autoimmune diseases are eligible for this study if they meet all of the following criteria:

[0613] The rash must cover <10% of the body surface area.

[0614] The disease is well controlled at baseline and requires only low-potency topical corticosteroids.

[0615] There has been no acute exacerbation of a potential condition requiring psoralen plus UVA radiation, methotrexate, retinoids, biologics, oral calcineurin inhibitors, or high-potency or oral corticosteroids within the past 12 months.

[0617] Malabsorption syndrome or other conditions that may affect intestinal absorption

[0618] Pregnancy or lactation, or planning to become pregnant during the study period or within 12 months after the last dose of R-CHOP, within 3 months after the last dose of tocilizumab (if applicable), or within 2 months after the last dose of glimepiride.

[0619] Endpoints

[0620] The primary efficacy endpoint is the EOT CR rate, as measured by PET-CT.Scan measurements were performed and determined by the investigator according to the 2014 Lugano response criteria. The CR rate was estimated as the percentage of participants who achieved CR at EOT, and a corresponding Clopper-Pearson precision 95% CI was constructed for the study cohort.

[0621] Secondary endpoints

[0622] The secondary efficacy endpoints were EOT ORR, PFS, and OS.

[0623] ORR was defined as CR or PR at EOT, measured by PET-CT scans and determined by the investigator. The analysis of ORR was the same as that described above for the primary efficacy endpoint of CR rate at EOT measured by PET / CT scans.

[0624] PFS was defined as the time from the first study treatment to the first occurrence of disease progression, relapse, or death from any cause (whichever occurs first). For participants who did not experience disease progression, relapse, or death, PFS was reviewed on the date of the last evaluable tumor assessment. For participants with no baseline evaluable tumor assessment, PFS was reviewed one day after the date of the first study treatment. PFS was assessed by the investigators using the 2014 Lugano Response Criteria (Cheson et al. 2014). The distribution of PFS, median (if analytically feasible) 6-month and 1-year PFS were estimated using the Kaplan-Meier method (Kaplan and Meier 1958), with the 95% CI for median PFS constructed using the Brookmeyer-Crowley method (Brookmeyer and Crowley 1982). The standard errors and corresponding 95% CIs for 6-month and 1-year PFS were calculated using the Greenwood formula.

[0625] OS was defined as the time from the first treatment in the study to death from any cause. The analysis of OS was the same as that described above for PFS.

[0626] Relevant laboratory, vital signs (pulse rate, respiratory rate, blood pressure and temperature), and ECG data were presented over time and graded where appropriate. In addition, a transposed table of selected laboratory tests was used to summarize the severity grades at baseline and maximum baseline. Changes in vital signs and ECG will be summarized.

[0627] Safety is assessed by summarizing study treatment exposures, occurrence and severity of adverse events, changes in laboratory test results, and changes in vital signs and ECG. Study treatment exposures (such as duration of treatment, total dose received, number of cycles, and number of dose modifications) are summarized using descriptive statistics. All verbatim adverse event terms are mapped to MedDRA terminology and adverse event severity is graded according to NCI CTCAE v5.0. [Note: The last sentence appears to be incomplete and possibly refers to a separate section.]Page 53 / 71, CN 122070142 A. In CRS, severity was determined according to the ASTCT CRS consensus grading criteria. All adverse events, critical adverse events, adverse events leading to death, adverse events of particular concern, and adverse events leading to termination of study treatment (i.e., adverse events occurring during treatment) occurring during or after the first administration of study treatment were summarized using mapped terms, appropriate thesaurus levels, and severity grades. For events of different severity, the highest grade was used in the summary. Deaths and causes of death were summarized.

[0628] Exploratory Endpoints

[0629] Exploratory efficacy endpoints were DOR and EOT CR rate, measured by PET / CT scans and determined by automated response (aLugano) according to the 2014 Lugano Response Criteria. DOR was defined as the time from the first recorded objective response (CR or PR) to disease progression or death from any cause, whichever occurred first. For participants who did not die or experience disease progression, DOR was reviewed at the date of the last evaluable oncology assessment. DOR was determined by the investigator using the 2014 Lugano response criteria (Cheson et al. 2014). DOR analysis only included participants who achieved an objective response (CR or PR) at any time during the study. The distribution of DOR and median DOR was estimated using the Kaplan-Meier method (Kaplan and Meier 1958) (where analytically feasible), and the 95% CI for median DOR was constructed using the Brookmeyer-Crowley method (Brookmeyer and Crowley 1982). DOR was pooled for participants with a best objective response of PR and subsets of participants with a best objective response of CR.

[0630] The aLugano analysis was performed in a PET / CT-evaluable population, defined as all patients with available PET / CT scans at baseline and during treatment. The analysis of EOT CR rates determined by aLugano was identical to the analysis described above for the primary efficacy endpoint of CR rates at EOT determined by the investigator.

[0631] A contingency matrix was calculated for aLugano and responses assessed according to the 2014 Lugano Response Criteria. The percentages of positive concordance, negative concordance, overall concordance, sensitivity, and specificity for CR, CR / PR, and DOR were calculated. Automated total metabolic tumor volume (aTMTV) was calculated for an evaluable patient population with baseline available PET / CT scans. The overall distribution of aTMTV at baseline and during treatment could be plotted using descriptive statistics (e.g., ...).Values ​​for the assessable population (e.g., mean, standard deviation, median, and range) can be summarized. Spearman correlation can be used to assess the consistency between aTMTV and TMTV as assessed by the sponsor's radiologist, and visualization can be performed using a scatter plot. The relationship between aTMTV and CR rate, ORR, PFS, OS, and DOR can be further assessed to determine whether the feature is prognostic or predictive. Additional analyses of quantitative measures (e.g., ΔSUVmax) and / or exploratory biomarkers can be performed within the scope of this study and after summarizing data from other studies.

[0632] Other safety analyses

[0633] Assess the incidence of ADA response (i.e., the presence of anti-glucantuma antibody in serum) and its potential correlation with PK, pharmacodynamic, and safety parameters.

[0634] Efficacy Assessment

[0635] Tumor and Response Assessment

[0636] Tumor assessment was performed on participants at screening, at the interim response assessment (between days 15 and 21 of cycle 2), and at EOT using 18FFDG-PET / CT and diagnostic CT scans. Response was assessed according to the 2014 Lugano Response Criteria. Response was assessed every 6 months during survival follow-up until any time the participant experienced disease progression, discontinued the study, suspected disease progression, or up to 1 year after the EOT response assessment, whichever occurred first. Tumor assessment was repeated at the discretion of the investigator if disease progression was suspected.

[0637] Clinical response assessment included evaluation of the presence and extent of lymphadenopathy, hepatomegaly, and splenomegaly via physical examination. All measurable and / or evaluable lesions were assessed and recorded at screening. Tumor assessments performed as SOCs before obtaining informed consent and within 28 days prior to the initiation of study treatment do not need to be repeated at screening, provided they meet the criteria outlined in the instructions below, page 54 / 71, 58 CN 122070142 A.

[0638] Radiographic Assessment

[0639] FDG-PET / CT scans combined with diagnostic quality CT scans are required at screening and EOT. Interim efficacy assessments (days 15 to 21 of cycle 2) require only FDG-PET / CT scans. At all times during the study period, diagnoses of disease progression based on clinical examination must be confirmed as soon as possible (within 30 days) and before initiation of non-protocol-prescribed anticancer therapies by imaging examinations (e.g., CT scans, FDG PET-CT scans). Bone marrow biopsy is not a routine requirement for response assessment, but may be indicated in specific circumstances according to the 2014 Lugano Response Criteria (e.g., response assessment based solely on CT or PET / CT).(When residual uptake is observed on the scan). All measurable and / or evaluable diseases must be recorded at screening and reassessed at each subsequent tumor assessment. Response is assessed by the investigator based on the results of physical examination, CT scan, and FDG-PET / CT scan.

[0640] Physical Examination

[0641] A complete physical examination includes at least assessments of the cardiovascular, skin, musculoskeletal, respiratory, gastrointestinal, and nervous systems; and, if clinically indicated, a genitourinary examination. Any abnormalities identified at baseline are recorded on the general history and baseline status eCRF. As part of a complete physical examination, the presence and extent of lymphadenopathy, hepatomegaly, and splenomegaly are recorded on the appropriate tumor assessment eCRF. A targeted, symptom-oriented physical examination includes at least assessments of the skin, respiratory system, cardiovascular system, and abdomen (liver and spleen) and is performed at the designated post-baseline visit and as indicated by clinical signs. The investigator must pay particular attention to clinical signs associated with previous serious diseases. Changes in abnormalities relative to baseline are recorded in the participant's notes. New or worsening clinically significant abnormalities are recorded as adverse events on the adverse event eCRF. Targeted physical examinations, as part of tumor assessment, include evaluation for the presence and extent of lymphadenopathy, hepatomegaly, splenomegaly, or other findings associated with lymphoma, and are recorded on the appropriate tumor assessment eCRF.

[0642] Sample Size Determination

[0643] The aim of this study is to evaluate the safety and efficacy of the combination of glimepiride and R-CHOP in a first-line setting in participants with previously untreated DLBCL at high ctDNA risk. This study is ongoing. Based on previously reported high ctDNA risk rates of approximately 20% in patients with DLBCL after one cycle of R-CHOP therapy in a first-line setting, approximately 200 participants will be screened to enroll 40 evaluable participants in the study treatment (Kurtz et al. 2018). The primary efficacy endpoint is the CR rate at EOT as assessed by the investigator according to the 2014 Lugano Response Criteria. For 40 patients, the error margin of the precise Clopper-Pearson (Clopper and Pearson 1934) 95% CI used to estimate the true CR rate at EOT was no more than ±17%. Based on a sample size of 40, the precise Clopper-Pearson 95% CI corresponded to observed CR rates ranging from 40% to 80%. It has been previously reported that the optimal CR rate was 40% for patients with high-risk ctDNA DLBCL receiving SOC R-CHOP therapy in a first-line setting (Kurtz et al. 2018). In 40...For example, if a participant has an observed CR rate of at least 60%, a true CR rate of less than 43% or more than 75% can be excluded with 95% confidence.

[0644] Results

[0645] We report preliminary safety and efficacy from a phase II open-label prospective study (NCT04980222) evaluating glofit + R-CHOP in patients with 1L LBCL who were defined as high risk based on ctDNA kinetics. Eligible patients had 1L CD20+ LBCL, with an Eastern Oncology Cooperative Group performance status of 0 to 2; IPI of 0 to 5 (IPI of 0 for patients with larger lesions; IPI of 2 to 5 for US patients). During ctDNA screening, patients received R-CHOP (21-day cycle) on day 1 (D) of cycle (C)1 and cycle C2. In patients defined as high-risk by ctDNA (plasma ctDNA decrease <2 log after one R-CHOP cycle) according to the instructions on page 55 / 71 of CN 122070142 A, R-CHOP continued to C6, with glimetuzumab administered in escalating doses at C3 (D8, 2.5 mg; D15, 10 mg), and at the target dose (30 mg) at D8 in C4–6 and D1 in C7–10 (21-day cycle). Hospitalization for cytokine release syndrome (CRS) was determined at the investigator's discretion based on the first dose of glimetuzumab; granulocyte colony-stimulating factor was recommended for neutropenia prophylaxis. Real-time ctDNA analysis was performed centrally (AVENIO oncology assay for non-Hodgkin lymphoma) (Stokowski et al. ASH 2022). The primary endpoint was complete response (CR) at the end of treatment (EOT). Responses were assessed by investigators via PET according to the Lugano criteria at C2 and EOT (Cheson et al., J Clin Oncol 2014). CRS was graded according to the ASTCT criteria (Lee et al., Biol Blood Marrow Transplant 2019); other adverse events (AEs) were assessed according to CTCAE v5.0. At the data cutoff date (May 25, 2023), of the 121 screened patients, 24 (19.8%) were defined as high-risk based on ctDNA (47 low-risk patients [38.8%]; 49 non-evaluable patients [40.5%]) and received ≥1 dose of the study drug from C3D1 (safety cohort).Of the 24 high-risk patients, 15 met the EOT assessment (14 completed treatment; 1 discontinued treatment due to disease progression [PD]), and 9 are still receiving treatment. Histological examination revealed no nonspecific high-grade B-cell lymphoma in any of the 24 patients; 2 had double-hit and 4 had triple-hit cytogenetic patterns; the median age was 61.5 years (range: 39–77); 14 (58.3%) patients had an IPI of 1–2 (IPI 1: 12.5%, IPI 2: 45.8%), and 9 (37.5%) patients had an IPI of 3–5 (IPI 3: 29.2%, IPI 4: 8.3%, IPI 5: 4.2%). In the safety cohort, 20 patients (83.3%) experienced any grade of adverse event (AE); 15 patients (62.5%) experienced grade 3 / 4 AEs, including neutropenia (11 patients [45.8%]); no febrile neutropenia was observed (Table 5). The most common (>20%) treatment-intervention AEs (any grade) were neutropenia (13 patients [54.2%]) and diarrhea (6 patients [25.0%]). No grade 5 AEs were reported. Nine patients (37.5%) experienced 17 serious AEs (14 of which were related to glimetuzumab), including 7 cases of CRS. Eight patients (33.3%) experienced ≥ grade 3 glimetuzumab-related AEs. Five patients (20.8%) experienced CRS events: four (16.7%) had grade 1 fever, and one (4.2%) had grade 2 fever and hypotension due to hypotension. CRS events occurred early and resolved by the data cutoff; tocilizumab and dexamethasone were administered only for grade 2 CRS events. One patient (4.2%) experienced a non-immunotoxic grade 3 neurological adverse event (syncope aura, unrelated to glimetuzumab), which resolved by the data cutoff. One patient died from PD during treatment follow-up after discontinuation of glimetuzumab. The interim CR rate was 46.7% (95% CI: 21.3–73.4); the CR rate at EOT was 80.0% (95% CI: 51.9–95.7), and the overall response rate was 93.3% (95% CI: 68.1–99.8) (Figure 4). Glofit + R-CHOP demonstrated a manageable safety profile in 1L high-risk LBCL as defined by ctDNA and induced high response rates at interim analysis and EOT. Dynamic treatment-interim risk assessment using ctDNA provided baseline-independent characteristics.To identify the potential of high-risk patients with LBCL.

[0646] Table 5: Summary of AEs (>10%) by highest NCI CTCAE grade. Instructions for Use, 56 / 71 pages, 60 CN 122070142 A

[0647]

[0648] * Neutropenia: Combined result of neutropenia and decreased neutrophil count; multiple occurrences of the same AE in the same individual are counted once by the highest grade of that AE.

[0649] AE, adverse event; CTCAE, Common Terminology Standard for Adverse Events; NCI, National Cancer Institute.

[0650] Updated Results

[0651] Table 6 below reports updated End of Treatment (EOT) results after another data cutoff date (September 18, 2023). The overall response rate was 95% (see “Responder” percentage). The secondary efficacy group of EoT includes all patients who received the first dose of its study treatment early enough (after enrollment) to be scheduled for treatment completion response assessment before the data cutoff date.

[0652]

[0653] Sequence Listing Specification 57 / 71 pages 61 CN 122070142 A

[0654] Sequence ID No:1

[0655] Length: 5

[0656] Molecular Type: Protein

[0657] Source: Artificial Sequence

[0658] Species: Synthetic Construct

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[0660]

[0661] Sequence ID No:2

[0662] Length: 17

[0663] Molecular Type: Protein

[0664] Source: Artificial Sequence

[0665] Species: Synthetic Construct

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[0668] Sequence ID No:3

[0669] Length: 10

[0670] Molecular Type: Protein

[0671] Source: Artificial Sequence

[0672] Species: Synthetic Construct

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[0675] Sequence ID No:4

[0676] Length: 16

[0677] Molecular type: Protein

[0678] Source: Artificial sequence

[0679] Species: Synthetic construct

[0680] Sequence:

[0681]

[0682] Sequence ID No:5

[0683] Length: 7

[0684] Molecular type: Protein

[0685] Source: Artificial sequence

[0686] Species: Synthetic construct

[0687] Sequence: Specification 58 / 71 pages 62 CN 122070142 A

[0688]

[0689] Sequence ID No:6

[0690] Length: 9

[0691] Molecular type: Protein

[0692] Source: Artificial sequence

[0693] Species: Synthetic construct

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[0695]

[0696] Sequence ID No:7

[0697] Length: 119

[0698] Molecular type: Protein

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[0700] Species: Synthetic construct

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[0703] Sequence ID No:8 Specification 59 / 71 Page 63 CN 122070142 A

[0704] Length: 112

[0705] Molecular type: Protein

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[0707] Species: Synthetic construct

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[0709]

[0710] Sequence ID No:9

[0711] Length: 5

[0712] Molecular type: Protein

[0713] Source: Artificial sequence

[0714] Species: Synthetic construct

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[0717]

[0718] Sequence ID No: 10

[0719] Length: 19

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[0723] Sequence: Specification 60 / 71 Page 64 CN 122070142 A

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[0725] Sequence ID No: 11

[0726] Length: 14

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[0732] Sequence ID No: 12

[0733] ​​Length: 14

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[0736] Species: Synthetic construct

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[0739] Sequence ID No: 13

[0740] Length: 7

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[0746] Sequence ID No: 14

[0747] Length: 9

[0748] Molecular type: Protein

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[0753] Sequence ID No: 15

[0754] Length: 125

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[0767] Sequence ID No: 17

[0768] Length: 672

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[0773] Specification 63 / 71 pages 67 CN 122070142 A Specification 64 / 71 pages 68 CN 122070142 A Specification 65 / 71 pages 69 CN 122070142 A

[0774] Sequence ID No: 18

[0775] Length: 447

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[0780] Specification 66 / 71 pages 70 CN 122070142 A Specification 67 / 71 pages 71 CN 122070142 A Specification 68 / 71 pages 72 CN 122070142 A

[0781] Sequence ID No: 19

[0782] Length: 232

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[0787] Specification 69 / 71 pages 73 CN 122070142 A

[0788] Sequence ID No: 20

[0789] Length: 219

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[0792] Species: Synthetic construct

[0793] Sequence:

[0794] Specification 70 / 71 pages 74 CN 122070142 A

[0795] Other embodiments

[0796] Although the invention has been described in considerable detail above by way of demonstration and example for the purpose of clear understanding, such description and example should not be construed as limiting the scope of the invention. All patent and scientific literature disclosures cited herein are expressly incorporated herein by reference in their entirety. Specification 71 / 71 pages 75 CN 122070142 A Figure 1 Specification Figure 1 / 4 pages 76 CN 122070142 A Figure 2 Specification Figure 2 / 4 pages 77CN 122070142 A Figure 3, Appendix to the Specification, Page 3 / 4, 78 CN 122070142 A Figure 4, Appendix to the Specification, Page 4 / 4, 79 CN 122070142 A

Claims

1. A method for treating a previously untreated patient with diffuse large B-cell lymphoma at high risk of circulating tumor DNA (ctDNA), the method comprising administering a combination of a bispecific antibody targeting CD3 and CD20, comprising the VH and VL sequences of glimetuzumab, with chemotherapy, wherein the patient has been identified as at high risk of ctDNA if the ctDNA level decreases by less than 2.5 log after one or two cycles of chemotherapy.

2. The method according to claim 1, wherein the bispecific antibody targeting CD3 and CD20 comprises the heavy chain of SEQ ID NO: 17, the heavy chain of SEQ ID NO: 18, the two light chains of SEQ ID NO: 20, and the light chain of SEQ ID NO: 19; and wherein the heavy chain and the light chain are assembled to form a first Fab molecule specifically binding to CD20, a second Fab molecule specifically binding to CD3, and a third Fab molecule specifically binding to CD20, and an Fc domain composed of a first subunit and a second subunit capable of stable association.

3. The method according to claim 1 or 2, wherein the chemotherapy is selected from: rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP); rituximab, cyclophosphamide, doxorubicin and prednisone (R-CHP); or rituximab, ifosfamide, carboplatin and etoposide phosphate (R-ICE).

4. The method according to any one of claims 1 to 3, wherein the bispecific antibody targeting CD3 and CD20 is glimetuzumab, and the chemotherapy is rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), and wherein the patient has been identified as high-risk for ctDNA if the patient's ctDNA decreases by less than 2.5 log after one or two R-CHOP cycles.

5. The method of claim 4, wherein the reduction in ctDNA is determined by measuring the amount of ctDNA before or on the day of the first treatment (baseline) and after at least one R-CHOP treatment cycle.

6. The method of claim 4 or 5, wherein the amount of ctDNA is determined on or before day 1 of the first R-CHOP treatment cycle (baseline) and on day 1 of the second R-CHOP treatment cycle.

7. The method according to any one of claims 4 to 6, wherein if the patient's ctDNA decreases by less than 2 log, the patient is identified as high-risk for ctDNA.

8. A method for treating previously untreated diffuse large B-cell lymphoma, the method comprising: a. Administer rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles; b. Determine the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and c. Select patients whose ctDNA reduction is <2.5-log between baseline and at least one R-CHOP treatment cycle and continue treatment with R-CHOP and glimetuzumab.

9. The method of claim 8, wherein step a) comprises applying R-CHOP on day 1 of cycles 1, 2 and optionally cycle 3.

10. The method of claim 9, wherein step a) comprises applying R-CHOP on day 1 of cycles 1 and 2.

11. The method according to any one of claims 8 to 10, wherein step b) comprises determining the amount of ctDNA on or before day 1 of cycle 1 (baseline) and day 1 of cycle 2 or day 1 of cycle 3 of the R-CHOP treatment in step a).

12. The method of claim 11, wherein step b) comprises determining the amount of ctDNA on day 1 (baseline) of cycle 1 and day 1 of cycle 2 of the R-CHOP treatment in step a).

13. The method according to any one of claims 8 to 12, wherein step c) comprises selecting patients with a ctDNA reduction of <2-log.

14. The method according to any one of claims 8 to 13, wherein step c) comprises continuing treatment with R-CHOP for a total of six cycles (cycles 1 to 6) and adding glimetuzumab for a total of eight cycles (cycles 3 to 10).

15. The method according to any one of claims 8 to 14, wherein glucentumab is administered at a dose of 2.5 mg on day 8 of cycle 3 and at a dose of 10 mg on day 15, at a dose of 30 mg on day 8 of cycles 4 to 6, and at a dose of 30 mg on day 1 of cycles 7 to 10.

16. The method according to any one of the preceding claims, wherein the ctDNA is circulating tumor DNA in plasma.

17. The method according to any one of the preceding claims, wherein the amount of ctDNA is measured in a blood sample from the patient.

18. The method of claim 17, wherein the amount of ctDNA is measured in genomic DNA isolated from peripheral mononuclear cells or plasma-depleted whole blood (PDWB) or in cell-free DNA (cfDNA) in plasma.

19. The method of claim 18, wherein the ctDNA is measured in cell-free DNA in plasma.

20. The method according to any one of the preceding claims, wherein the ctDNA is determined by targeted next-generation sequencing (NGS).

21. The method according to any one of claims 4 to 20, wherein rituximab is administered at 375 mg / m². 2 Cyclophosphamide was administered intravenously at a dose of 750 mg / m². 2 Doxorubicin was administered intravenously at a dose of 50 mg / m². 2 Vincristine was administered intravenously at a dose of 1.4 mg / m². 2 The dose was administered via IV bolus, and prednisone was administered at a dose of 100 mg / day from day 1 to day 5.

22. The method according to any one of claims 1 to 21, wherein the length of each cycle is 21 days.

23. The method according to any one of claims 4 to 22, wherein administration of such treatment to multiple human patients at high risk of ctDNA results in an overall response rate (ORR) of at least 80%, at least 85%, at least 90%, or at least 95%.

24. The method of claim 23, wherein the ORR is at least 95%.

25. The method according to any one of claims 1 to 21, wherein administration of such treatment to multiple human patients at high risk of ctDNA results in a complete remission (CR) rate of at least 75%, at least 80%, or at least 85%.

26. The method of claim 25, wherein the CR rate is at least 85%.

27. The method according to claims 23 and 24 or claims 25 and 26, wherein the ORR or CR rate is determined at the end of treatment.

28. The method of claim 27, wherein the ORR or CR is determined by PET-CT scan.

29. The method of claim 27 or 28, wherein the ORR or CR is determined according to the 2014 Lugano Reaction Standard.

30. A bispecific antibody targeting CD3 and CD20 comprising the VH and VL sequences of glimetuzumab, for use in a method of treating previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, the method comprising administering glimetuzumab in combination with chemotherapy to a patient, wherein the patient has been identified as at high risk of ctDNA if the ctDNA level decreases by less than 2.5 log after one cycle of chemotherapy.

31. Glimepiride, in a method of treating previously untreated diffuse large B-cell lymphoma with high (ct)DNA risk, the method comprising: a. Administer rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles; b. Determine the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and c. Select patients whose ctDNA reduction is <2.5-log between baseline and at least one R-CHOP treatment cycle and continue treatment with R-CHOP and glimetuzumab.

32. Use of a bispecific antibody targeting CD3 and CD20 comprising the VH and VL sequences of glimetuzumab in the manufacture of a medicament for treating a subject with previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, wherein the patient is administered glimetuzumab in combination with chemotherapy, wherein the patient has been identified as at high risk of ctDNA if the ctDNA decreases by less than 2.5 log after one cycle of chemotherapy.

33. Use of glimetuzumab in the manufacture of a medicament for the treatment of subjects with previously untreated diffuse large B-cell lymphoma at high risk of (ct)DNA, including: a. Administer rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) for at least 2 cycles; b. Determine the amount of ctDNA at or before cycle 1 (baseline) and after at least one R-CHOP treatment cycle; and c. Select patients whose ctDNA reduction is <2.5-log between baseline and at least one R-CHOP treatment cycle and continue treatment with R-CHOP and glimetuzumab.

34. The present invention as described above.

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