Pharmaceutical composition for treating diffuse large B-cell lymphoma and application thereof

Abstract

The invention discloses application of an anti-CD20 antibody drug conjugate or a drug combination thereof in preparation of a drug for treating diffuse large B-cell lymphoma. The drug combination comprises the anti-CD20 antibody drug conjugate, gemcitabine and oxaliplatin, the anti-CD20 antibody drug conjugate or the drug combination therapy containing the anti-CD20 antibody drug conjugate has a better curative effect compared with the existing clinical second-line standard therapy.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This patent application claims the priority benefit of the Chinese invention patent application with application number CN 202311225386.1 filed on September 21, 2023, the entire contents of which are hereby incorporated by reference. Technical Field

[0003] The present invention relates to the technical field of pharmaceutical and drug combination, and in particular to a drug combination for treating diffuse large B-cell lymphoma and a use thereof. Background Art

[0004] Non-Hodgkin lymphoma (NHL) is the seventh most common type of cancer in the world, accounting for approximately 4% to 5% of new cancer cases and 3% to 4% of cancer-related deaths, affecting approximately 1.5 million people worldwide.

[0005] NHL is a general term for a group of malignant proliferative diseases of the lymphatic system, including many types of lymphocytic neoplasms, which mainly originate from B lymphocytes (>85%), and a few originate from T cells and natural killer cells. It is usually classified according to the prognosis into slowly progressing or indolent NHL (iNHL) and rapidly progressing or aggressive NHL (aNHL); among them, follicular lymphoma (FL) is the most common iNHL, accounting for about 1 / 5 of all NHL patients, while diffuse large B-cell lymphoma (DLBCL) is the most common aNHL, accounting for about 1 / 3 of all NHL patients.

[0006] Among DLBCL, 30%-35% express MYC protein, and 20-35% also express BCL2, which is called "double-expressing lymphoma", indicating a poor prognosis.

[0007] Combining drugs to improve disease treatment outcomes is a common approach in this field. However, the standard drug combination treatment regimens used for different lines and subtypes of non-Hodgkin's lymphoma vary clinically. There are even more drug combination regimens under study, and the types of chemical and biological drugs that can be used in combination are even more complex, making the design of drug combination regimens very difficult, and there is great uncertainty about the efficacy and safety of drug combinations.

[0008] WO2021223048A1 and its Chinese homologue CN115485304A disclose an anti-CD20 antibody-drug conjugate, and mention in the specification that the conjugate can be used in combination with doxorubicin, cyclophosphamide, taxanes, capecitabine, gemcitabine, irinotecan, oxaliplatin, prednisone, vincristine, etc., or a combination thereof; the applicant of the present invention previously applied for CN108452318A and CN108452319A, respectively disclosing an anti-CD20 antibody-drug conjugate and its preparation, and mention in the specification that the anti-CD20 antibody-drug conjugate can be used in combination with cyclophosphamide, doxorubicin, vincristine, prednisone, PD1 antibody, CTLA4 inhibitor, or a combination thereof.

[0009] However, the above patent application simply lists the combination of anti-CD20 conjugates and commonly used anti-tumor drugs in clinical practice. The examples do not design specific drug combination schemes for the treatment of non-Hodgkin's lymphoma and obtain efficacy and safety data. Moreover, the above-mentioned taxanes, PD1 antibodies, CTLA4 inhibitors and other drugs used in combination with anti-CD20 antibody-drug conjugates have not even been used in the treatment of non-Hodgkin's lymphoma in clinical practice. The listing of various drugs used in combination in the above patents is only a way of writing patents and is accidental disclosure. Due to the variety of drugs available for combination, the combination of combination schemes, and the uncertainty of the efficacy of drug combination and the superposition of toxicity of combined drugs, the above patent application cannot provide technical inspiration for specific drug combination schemes.

[0010] The first-line standard treatment regimens for DLBCL currently recommended by the Chinese Society of Clinical Oncology (CSCO) Lymphoma Diagnosis and Treatment Guidelines 2022 are shown in Table a below, including R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R, and Pola-R-CHP. The R-CHOP regimen uses rituximab (R), cyclophosphamide (C), vincristine (H), doxorubicin (O), and prednisone (P). The R-CHOEP regimen uses rituximab (R), cyclophosphamide (C), and vincristine (H). Doxorubicin (O) and prednisone (P) are combined. ), vincristine (H), doxorubicin (O), etoposide (E) and prednisone (P); R-miniCHOP regimen uses rituximab (R), cyclophosphamide (C), vincristine (H), doxorubicin (O) and prednisone (P), among which CHOP uses a low dose; DA-EPOCH-R uses etoposide (E), prednisone (P), doxorubicin (O), cyclophosphamide (C), vincristine (H) and rituximab (R), and after each chemotherapy cycle, the dose is adjusted according to the patient's response to chemotherapy (Dose Adjustment, DA); Pola-R-CHP uses Polatuzumab vedotin (ADC targeting CD79b, trade name ), rituximab (R), cyclophosphamide (C), vincristine (H) and prednisone (P).

[0011] Table a First-line treatment options for DLBCL

[0012]

[0013] Although patients have achieved varying degrees of relief through the above-mentioned first-line therapies, 40%-50% are still prone to relapse or refractory disease, especially for "double-expression lymphoma", which shows a poor prognosis.

[0014] The second-line standard treatment options for relapsed or refractory DLBCL after first-line standard treatment are currently recommended by the Chinese Society of Clinical Oncology (CSCO) Lymphoma Diagnosis and Treatment Guidelines 2022 as shown in Table b below, including R-GemOx (rituximab (R), gemcitabine (Gem) and oxaliplatin (Ox) combination), R 2 (combination of rituximab (R) and lenalidomide (R)), BR (combination of bendamustine (B) and rituximab (R)), R-ICE (combination of rituximab, ifosfamide, carboplatin and etoposide), R-DHAP (combination of rituximab, cisplatin, cytarabine and dexamethasone), R-GDP (combination of rituximab, gemcitabine, cisplatin and dexamethasone), R-MINE (combination of rituximab, mesna, ifosfamide, mitoxantrone and etoposide), R-ESHAP (combination of rituximab, etoposide, methylprednisolone, cisplatin and cytarabine), DA-EPOCH-R (combination of rituximab, etoposide, prednisone, vincristine, cyclophosphamide and doxorubicin).

[0015] Table b Second-line treatment options for DLBCL

[0016]

[0017] Although patients have achieved a certain degree of relief through the above-mentioned second-line standard treatment regimen, there is still room for further improvement in clinical benefits. Therefore, there is still a need to find more effective treatments or combination treatment drugs and treatment regimens for diffuse large B-cell lymphoma than existing standard treatments. Summary of the invention

[0018] The object of the present invention is to provide a drug combination and its use in the preparation of a drug for treating non-Hodgkin's lymphoma, wherein the drug combination combined therapy has a better therapeutic effect than the existing clinical second-line standard therapy.

[0019] The first aspect of the present invention discloses the use of an anti-CD20 antibody-drug conjugate in a drug for treating non-Hodgkin's lymphoma.

[0020] The anti-CD20 antibody drug conjugate has the following structural formula:

[0021] mAb-(LD)n;

[0022] Wherein, mAb represents recombinant anti-CD20 monoclonal antibody, D represents small molecule toxin; L is a linker connecting the monoclonal antibody and the small molecule toxin; n is the average coupling number of the small molecule toxin coupled to the monoclonal antibody; and “-” represents a bond.

[0023] Preferred recombinant anti-CD20 monoclonal antibodies include but are not limited to rituximab or its biosimilars, ofatumumab or its biosimilars, obinutuzumab or its biosimilars, ocrelizumab or its biosimilars.

[0024] More preferably, the recombinant anti-CD20 monoclonal antibody is rituximab or a biosimilar thereof.

[0025] Preferably, D is one or more monomethyl auristatins, wherein the monomethyl auristatin includes but is not limited to monomethyl auristatin-E (MMAE), monomethyl auristatin-D (MMAD) or monomethyl auristatin-F (MMAF); further, the monomethyl auristatin is monomethyl auristatin-E (MMAE).

[0026] L is a cleavable linker or a non-cleavable linker, wherein the non-cleavable linker includes but is not limited to a thioether bond linker, wherein the thioether bond linker comprises a maleimide group, and preferably the maleimide group also comprises hexanoic acid to form a maleimidocaproyl group (mc); preferably the maleimide group comprises a maleimidomethyl group, such as succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sMCC) or sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-sMCC).

[0027] The cleavable linker includes but is not limited to a hydrazone linker, a disulfide linker, and a peptide linker; the peptide linker includes valine-citrulline (Val-Cit or VC), phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala) or valine-lysine (Val-Lys). The preferred peptide linker is valine-citrulline (Val-Cit).

[0028] Furthermore, the linker portion further comprises p-aminobenzyl alcohol (PAB), p-aminobenzyloxycarbonyl (PABC) or derivatives or analogs thereof.

[0029] Furthermore, L is preferably MC-VC-PAB.

[0030] Said n is an integer or non-integer of 1-8; preferably n is 4.2±1, more preferably 4.2±0.5, and further preferably 4.2±0.3.

[0031] The anti-CD20 antibody drug conjugate further has the following structure:

[0032]

[0033] Wherein, mAb is rituximab or a biosimilar thereof, and the average coupling number n of the small molecule toxin coupled to the antibody is 1-8; preferably n is 4.2±1, further preferably n is 4.2±0.5, and further preferably n is 4.2±0.3.

[0034] The anti-CD20 antibody-drug conjugate of the present invention may be a pharmaceutical preparation containing the anti-CD20 antibody-drug conjugate and a carrier or an excipient, and the pharmaceutical preparation is a liquid preparation or a lyophilized preparation.

[0035] The carrier or excipient is selected from the following group: pH buffer, osmotic pressure regulator, lyophilized powder excipient, protein protectant, solubilizer, water for injection.

[0036] The pH buffer includes histidine hydrochloride buffer, acetate buffer, phosphate buffer, citrate buffer or Tris buffer, and preferably the pH buffer is histidine hydrochloride buffer.

[0037] The protein protective agent includes trehalose, sucrose, lactose or glucose, and preferably the protein protective agent is trehalose.

[0038] The osmotic pressure regulator includes mannitol or sodium chloride, and preferably the osmotic pressure regulator is mannitol.

[0039] The solubilizer includes polysorbate 80, polysorbate 20 or poloxamer 188, and preferably the solubilizer is polysorbate 80.

[0040] Furthermore, the liquid or lyophilized preparation comprises the above-mentioned anti-CD20 antibody drug conjugate, histidine hydrochloride, trehalose, mannitol and polysorbate 80.

[0041] Preferably, the anti-CD20 antibody-drug conjugate is used in combination with at least one therapeutic agent, and the at least one therapeutic agent is selected from antimetabolite drugs and platinum drugs.

[0042] The antimetabolite drug is selected from one or more of pemetrexed, methotrexate, fluorouracil, capecitabine, cytarabine, gemcitabine, 6-mercaptopurine and thioguanine;

[0043] The platinum drug is selected from one or more of carboplatin, cisplatin and oxaliplatin.

[0044] Furthermore, the at least one therapeutic agent is gemcitabine and oxaliplatin.

[0045] Preferably, the non-Hodgkin's lymphoma is selected from small lymphocytic lymphoma, B-cell chronic lymphocytic leukemia, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, mantle cell lymphoma, precursor (peripheral) large granular lymphocytic leukemia or anaplastic large cell lymphoma.

[0046] Preferably, the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.

[0047] Preferably, the diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma.

[0048] Furthermore, the diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma after receiving at least one standard treatment regimen.

[0049] Furthermore, the diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

[0050] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with dual expression of MYC and BCL2.

[0051] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with high CD20 expression or low CD20 expression.

[0052] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with high expression of CD20 and high expression of one or more of MYC, BCL2 and BCL6.

[0053] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with high expression of CD20 and high expression of MYC and BCL2.

[0054] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with low expression of CD20 and high expression of one or more of MYC, BCL2 and BCL6.

[0055] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with low expression of CD20 and high expression of MYC and BCL2.

[0056] Preferably, the at least one standard treatment regimen includes at least anti-CD20 antibody monotherapy or anti-CD20 antibody combination therapy.

[0057] Further, the at least one standard treatment regimen includes at least one selected from R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R and Pola-R-CHP.

[0058] Furthermore, the at least one standard treatment regimen is R-CHOP.

[0059] The second aspect of the present invention discloses a drug combination for treating non-Hodgkin's lymphoma

[0060] The drug combination comprises an anti-CD20 antibody drug conjugate and at least one therapeutic agent, wherein the anti-CD20 antibody drug conjugate has the following structure:

[0061]

[0062] Herein, mAb represents a recombinant anti-CD20 monoclonal antibody, the drug-antibody ratio DAR is 1-8, the preferred DAR is 4.2±1, the further preferred DAR is 4.2±0.5, and the further preferred DAR is 4.2±0.3.

[0063] Preferably, the recombinant anti-CD20 monoclonal antibody is selected from rituximab or its biosimilars, ofatumumab or its biosimilars, ocrelizumab or its biosimilars, and obinutuzumab or its biosimilars.

[0064] Further preferably, the recombinant anti-CD20 monoclonal antibody is selected from rituximab or a biosimilar thereof.

[0065] Preferably, the at least one therapeutic agent is selected from antimetabolites and platinum drugs; the antimetabolites are selected from one or more of pemetrexed, methotrexate, fluorouracil, capecitabine, cytarabine, gemcitabine, 6-mercaptopurine and thioguanine; the platinum drugs are selected from one or more of carboplatin, cisplatin and oxaliplatin.

[0066] Furthermore, the at least one therapeutic agent is gemcitabine and oxaliplatin.

[0067] Preferably, the drug combination of the present invention is a combination of the anti-CD20 antibody-drug conjugate, gemcitabine and oxaliplatin.

[0068] The above-mentioned combination of the anti-CD20 antibody-drug conjugate and at least one therapeutic agent is only an exemplary combination and does not constitute a limitation on the combined use of the anti-CD20 antibody-drug conjugate of the present invention and other drugs.

[0069] The drug combination of the present invention consists of a first drug composition comprising the anti-CD20 antibody drug conjugate and a carrier or excipient thereof and a second drug composition comprising at least one therapeutic agent and a carrier or excipient thereof.

[0070] The first pharmaceutical composition and the second pharmaceutical composition of the present invention can be packaged independently or together.

[0071] The present invention also discloses a kit, which comprises the anti-CD20 antibody-drug conjugate and the at least one therapeutic agent.

[0072] Preferably, the anti-CD20 antibody drug conjugate and the at least one therapeutic agent are packaged independently and provided in the kit.

[0073] Preferably, the kit comprises the anti-CD20 antibody drug conjugate, gemcitabine and oxaliplatin.

[0074] The third aspect of the present invention discloses a combination therapy for treating non-Hodgkin's lymphoma.

[0075] The combined therapy of the present invention comprises administering an effective amount of the drug combination disclosed in the second aspect of the present invention for treating non-Hodgkin's lymphoma.

[0076] Preferably, the combination therapy comprises the combined administration of an effective amount of the anti-CD20 antibody drug conjugate, gemcitabine and oxaliplatin.

[0077] Preferably, the non-Hodgkin's lymphoma is selected from small lymphocytic lymphoma, B-cell chronic lymphocytic leukemia, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, mantle cell lymphoma, precursor (peripheral) large granular lymphocytic leukemia or anaplastic large cell lymphoma.

[0078] Preferably, the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.

[0079] Preferably, the diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma.

[0080] Furthermore, the diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma after receiving at least one standard treatment regimen.

[0081] Furthermore, the diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

[0082] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with dual expression of MYC and BCL2.

[0083] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with high CD20 expression or low CD20 expression.

[0084] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with high expression of CD20 and high expression of one or more of MYC, BCL2 and BCL6.

[0085] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with high expression of CD20 and high expression of MYC and BCL2.

[0086] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with low expression of CD20 and high expression of one or more of MYC, BCL2 and BCL6.

[0087] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with low expression of CD20 and high expression of MYC and BCL2.

[0088] Preferably, the at least one standard treatment regimen includes at least anti-CD20 antibody monotherapy or anti-CD20 antibody combination therapy.

[0089] Further, the at least one standard treatment regimen includes at least one selected from R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R and Pola-R-CHP.

[0090] Furthermore, the at least one standard treatment regimen is R-CHOP.

[0091] The anti-CD20 antibody drug conjugate is administered before, simultaneously with, and / or after the at least one therapeutic agent.

[0092] The anti-CD20 antibody drug conjugate and the at least one therapeutic agent can be administered once or multiple times. For example, the anti-CD20 antibody drug conjugate can be administered once or multiple times. For multiple administrations, the administration frequency can be, for example, four times a week, three times a week, twice a week, once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every seven weeks, or once every eight weeks, etc.; or one day, one week, two weeks, three weeks, four weeks, one month, two months, three months, six months as a cycle, and each cycle is administered once, twice, three times, four times, five times, six times, etc., and the administration is continued according to the appropriate administration cycle and frequency until the subject is cured, the disease symptoms progress, or the subject dies.

[0093] For at least one therapeutic agent, such as gemcitabine, oxaliplatin, etc., a single administration or multiple administration can be performed. For multiple administration, the administration frequency can be, for example, three times a day, twice a day, once a day, once every two days, once a week, once every two weeks, once every three weeks, once every four weeks, etc.; or one day, one week, two weeks, three weeks, four weeks, one month, two months, three months, six months as a cycle, and each cycle is administered once, twice, three times, four times, five times, six times, etc., and the administration is continued according to the appropriate administration cycle and frequency until the subject is cured, the disease symptoms progress, or the subject dies.

[0094] Regarding the dosage, depending on the type and severity of the disease, the initial candidate dosage of the anti-CD20 antibody drug conjugate administered to the patient can be about 1 μg / kg-100 mg / kg, and an exemplary dosage range of the anti-CD20 antibody drug conjugate can be, for example, 0.01 mg / kg-50 mg / kg, preferably about 0.05 mg / kg to about 20 mg / kg, preferably about 0.1 mg / kg to about 10 mg / kg, preferably about 0.5 mg / kg to about 5.0 mg / kg, and a preferred dosage is 0.5 mg / kg, 0.6 mg / kg, 0.75 mg / kg, 0.8 mg / kg, 1.0 mg / kg, 1.2 mg / kg, 1.5 mg / kg, 1.8 mg / kg, 2.0 mg / kg or 2.5 mg / kg.

[0095] Regarding the dosage, depending on the type and severity of the disease, the dosage of at least one therapeutic agent administered to the patient may be 1 mg / m 2 -100g / m 2 Or 1μg / kg-100mg / kg. For example, gemcitabine can be administered at a dose of 10mg / m 2 -100g / m 2 , preferably 100 mg / m 2 -10g / m 2 , preferably 200mg / m 2-5000mg / m 2 The preferred dosage is 250 mg / m 2 , 500mg / m 2 , 750mg / m 2 , 1000mg / m 2 , 1500mg / m 2 , 2000mg / m 2 , 2500mg / m 2 For example, oxaliplatin can be administered at a dose of 0.1 mg / m 2 -10g / m 2 Or 0.1μg / kg-10mg / kg; preferably, the dosage can be 1mg / m 2 -10g / m 2 , preferably 10 mg / m 2 -1000mg / m 2 , preferably 20 mg / m 2 -500mg / m 2 The preferred dosage is 25 mg / m 2 , 50mg / m 2 , 75mg / m 2 , 100mg / m 2 , 150mg / m 2 , 200mg / m 2 , 250mg / m 2 wait.

[0096] The combination drug of the present invention can be administered by any suitable means, including enteral, parenteral, intrapulmonary and intranasal administration, etc. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration, etc.

[0097] The fourth aspect of the present invention discloses a method for inhibiting the growth of non-Hodgkin's lymphoma cells.

[0098] The growth inhibition method comprises adding an effective dose of the anti-CD20 antibody-drug conjugate of the present invention or the drug combination of the second aspect of the present invention to the non-Hodgkin's lymphoma cell system.

[0099] One of the growth inhibition methods comprises adding an effective dose of an anti-CD20 antibody-drug conjugate to the non-Hodgkin's lymphoma cell system, wherein the anti-CD20 antibody-drug conjugate has the following structure:

[0100]

[0101] Herein, mAb represents a recombinant anti-CD20 monoclonal antibody, the drug-antibody ratio DAR is 1-8, the preferred DAR is 4.2±1, the further preferred DAR is 4.2±0.5, and the further preferred DAR is 4.2±0.3.

[0102] Preferably, the recombinant anti-CD20 monoclonal antibody is selected from rituximab or a biosimilar thereof.

[0103] Preferably, the DAR is 4.2±0.5.

[0104] The second growth inhibition method comprises administering an effective dose of the drug combination described in the second aspect of the present invention to the non-Hodgkin's lymphoma cell system.

[0105] Preferably, the growth inhibition method comprises administering an effective dose of the anti-CD20 antibody drug conjugate, gemcitabine and oxaliplatin to the non-Hodgkin's lymphoma cell system.

[0106] Preferably, the non-Hodgkin's lymphoma is selected from small lymphocytic lymphoma, B-cell chronic lymphocytic leukemia, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, mantle cell lymphoma, precursor (peripheral) large granular lymphocytic leukemia or anaplastic large cell lymphoma.

[0107] Preferably, the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.

[0108] Preferably, the diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma.

[0109] Furthermore, the diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma after receiving at least one standard treatment regimen.

[0110] Furthermore, the diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

[0111] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with dual expression of MYC and BCL2.

[0112] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with high CD20 expression or low CD20 expression.

[0113] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with high expression of CD20 and high expression of one or more of MYC, BCL2 and BCL6.

[0114] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with high expression of CD20 and high expression of MYC and BCL2.

[0115] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with low expression of CD20 and high expression of one or more of MYC, BCL2 and BCL6.

[0116] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with low expression of CD20 and high expression of MYC and BCL2.

[0117] Preferably, the at least one inhibitory treatment comprises at least an anti-CD20 antibody monotherapy or an anti-CD20 antibody combined with at least one therapeutic agent.

[0118] Furthermore, the at least one inhibition treatment includes at least one selected from R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R and Pola-R-CHP.

[0119] Furthermore, the at least one inhibitory treatment is R-CHOP.

[0120] Based on actual clinical needs, the present invention uses clinical standard treatment regimens as controls. For diffuse large B-cell lymphoma, especially diffuse large B-cell lymphoma with dual expression of BCL2 and MYC, the anti-CD20 antibody-drug conjugate combined with the Gemox regimen of the present invention shows better tumor inhibition effect and better treatment potential than the existing second-line standard therapy, while having good safety.

[0121] It should be understood that one, some or all of the various embodiments described herein can be combined to form other embodiments of the present invention. These and other embodiments of the present invention are further described by the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0122] The embodiments of the present invention are described in detail below with reference to the accompanying drawings, wherein:

[0123] Figure 1 : Change trend of tumor volume in mice of each treatment group in WSU-DLCL2 model;

[0124] Figure 2 : In the WSU-DLCL2 model, at the end of the experiment, the tumor weight of mice in each treatment group;

[0125] Figure 3 : Change trend of tumor volume in mice of each treatment group in OCI-LY19 model. DETAILED DESCRIPTION

[0126] The present invention is described below with reference to specific examples. It will be appreciated by those skilled in the art that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention in any way.

[0127] The experimental methods in the following examples are conventional methods unless otherwise specified. The raw materials, reagents, etc. used in the following examples are commercially available products unless otherwise specified.

[0128] definition:

[0129] "Rituximab" refers to a monoclonal antibody drug targeting CD20 developed by Roche and approved for marketing by drug regulatory authorities under the trade name "MabThera". Rituximab is its generic name, and only original research drugs can be called rituximab.

[0130] "Rituximab biosimilars" refer to: in addition to the original rituximab, other companies have developed monoclonal antibody drugs that imitate rituximab and have the same amino acid sequence as rituximab and similar physical and chemical properties, efficacy, pharmacokinetics and safety as the original rituximab.

[0131] "High expression" means that technicians identify the antigen or gene high expression as higher than a certain expression level or relatively high expression level based on the test results of tumor tissue or cell antigen or gene expression levels according to conventional evaluation standards. For example, strong positive (+++) and / or positive (++) determined by the proportion of immunohistochemically stained cells and the staining intensity can be identified as high expression of the antigen or gene.

[0132] "Low expression" means that technicians identify the antigen or gene low expression as below a certain expression level or a relatively low expression level based on the conventional evaluation standards based on the test results of tumor tissue or cell antigen or gene expression levels. For example, weak positive (+) determined by the proportion of immunohistochemically stained cells and staining intensity can be identified as low antigen or gene expression.

[0133] An "effective dose" refers to an amount effective to achieve the desired therapeutic result, at dosages and for periods of time necessary. The effective dose may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the therapeutic agent or combination of therapeutic agents to elicit a desired response in the individual.

[0134]

[0066] While the present invention has been generally described, embodiments of the present invention will be further disclosed in the following examples, which should not be construed as limiting the scope of the claims.

[0135] Example 1: Preparation of anti-CD20 ADC (TRS005)

[0136] The preparation method of the exemplary anti-CD20 ADC (TRS005) is specifically described in Chinese patent CN108452318B, which is fully incorporated into the present invention.

[0137] The structure of TRS005 is as follows:

[0138]

[0139] Among them, mAb was rituximab or rituximab biosimilar, and the drug-antibody ratio (DAR) was 4.2±0.5.

[0140] Example 2: Human lymphoma cell line screening

[0141] The expression levels (mRNA levels) of BCL2, MYC and CD20 in human diffuse large B-cell lymphoma cell lines SU-DHL-4, SU-DHL-6, TMD-8, WSUDLCL2, DOHH2, DB, OCI-LY10 and OCI-LY19 were detected by sequencing, and the human diffuse large B-cell lymphoma cell line WSU-DLCL2 with dual expression of BCL2 and MYC and high expression of CD20, and the human diffuse large B-cell lymphoma cell line OCI-LY19 with dual expression of BCL2 and MYC and low expression of CD20 were screened out. WSU-DLCL2 and OCI-LY19 are CD20-positive cell lines with dual expression of BCL2 and MYC. The expression levels of CD20, BCL2 and MYC in the two cell lines are shown in Table 1.

[0142] Table 1: Expression levels of CD20, BCL2 and MYC in two cell lines (log2TPM)

[0143] WSU DLCL2 OCI-LY19 CD20 9.360825 3.690417 BCL2 6.178316 5.953964 MYC 7.047015 9.049576

[0144] Example 3: Evaluation of the anti-tumor efficacy of TRS005 alone or in combination with Gemox in the WSU-DLCL2 / R-CHOP relapse model of human lymphoma

[0145] (I) Model establishment

[0146] The recurrence model was established by conventional operation methods in the art, such as inoculating WSU-DLCL2 cells into mice and treating them with R-CHOP, which was defined as P1; selecting tumor tissue with good growth status of WSU-DLCL2 / R-CHOP (P1), removing the mouse fascia and internal necrotic tissue, inoculating mice with plugs under sterile conditions, and treating them with R-CHOP, which was defined as P2; selecting tumor tissue with good growth status of WSU-DLCL2 / R-CHOP (P2), removing the mouse fascia and internal necrotic tissue, inoculating mice with plugs under sterile conditions, and treating them with R-CHOP, which was defined as P3; through R-CHOP treatment and passage from P1 to P3, an R-CHOP recurrence model was constructed.

[0147] The WSU-DLCL2 / R-CHOP (P3) tumor tissue with good growth status was selected and cut into 2×2×2mm pieces after removing the mouse fascia and internal necrotic tissue. 3 Small pieces were stored in RPMI 1640 basal medium for future use. After the animals were anesthetized with isoflurane, the tissue pieces were inoculated subcutaneously on the right side of the mouse using an 11G trocar under sterile conditions.

[0148] (II) Grouping and drug administration

[0149] When the average tumor volume reaches 150 mm 3 Around 24 hours, the animals were randomly divided into groups according to the tumor volume, so that the difference in tumor volume between groups was less than 10% of the mean. In this experiment, there were 6 animals in each group, and the day of grouping was recorded as Day 0. After grouping, the animals were immediately dosed according to their weight. On Day 24, the remaining animals were killed, the tumors were weighed and photographed, and the experiment ended. The grouping and dosing schedule are shown in Table 2-3.

[0150] Table 2. Dosing schedule for days 0-13

[0151]

[0152] Among them, IV means intravenous injection; IP means intraperitoneal injection; PO means oral administration; QW×2w means once a week for two weeks; BIW×2w means twice a week for two weeks; QD×5 / w×2w means once a day, 5 days a week, for two weeks.

[0153] Table 3. Dosing regimen for days 14-20

[0154]

[0155] On days 14-20, the dosing regimen of group 3 was changed to TRS005-Gemox, and the dosing regimens of other groups remained unchanged on days 14-20.

[0156] Animal body weight and tumor volume were measured twice a week during the experiment. Animal clinical symptoms were observed and recorded once a day during the experiment. Clinical observations included the overall health status of the animals, abnormal body weight, abnormal behavior, and other adverse reactions related to administration.

[0157] (III) Evaluation indicators and statistical analysis

[0158] Tumor growth inhibition rate (%TGI TV )The calculation formula is: (1-TV T / TV C )×100%,TV C is the average tumor volume of the negative control group, TV T is the average tumor volume of the treatment group.

[0159] The relative tumor volume (RTV) was calculated as follows: Vt / V0, where V0 was the tumor volume at the time of grouping and Vt was the tumor volume at each measurement.

[0160] Relative tumor proliferation rate (%T / C RTV ) is calculated as: RTV / C RTV × 100%, where T RTV RTV for the treatment group, C RTV RTV was the negative control group.

[0161] Tumor inhibition rate (%TGI TW )The calculation formula is: (TW C -TW T ) / TW C × 100%, of which TW C is the average tumor weight of the negative control group, TW T is the average tumor weight of the treatment group.

[0162] The calculation formula of animal body weight change rate (%BWC) is: (BWt-BW0) / BW0×100%, where BWt is the animal weight at each measurement and BW0 is the animal weight at grouping.

[0163] According to the China NMPA "Technical Guidelines for Non-clinical Studies of Cytotoxic Anti-tumor Drugs" (November 2006), %T / C RTV ≤40% and P<0.05 after statistical analysis was considered effective. If the number of drug-related animal deaths exceeded 20%, the drug dose was considered to be severely toxic.

[0164] The experimental data were expressed as Mean ± SEM, and the two-tailed t-test was used for comparison among the groups. P < 0.05 was considered a significant difference, and P < 0.01 was considered an extremely significant difference (Microsoft Excel 2007, Redmond, WA, USA).

[0165] (IV) Test results

[0166] 1. Anti-tumor effect of the test substance

[0167] In this experiment, the results of the study on the anti-tumor activity of TRS005 alone or in combination with Gemox in the relapse model of human lymphoma WSU-DLCL2 / R-CHOP are shown in Tables 4-5 and Figure 1-2 .

[0168] See Table 4 and Figure 1-2 On Day 24, the tumor weight of group 2R+Gemox group was 0.705±0.199g, and the tumor inhibition rate was 66.82%; the tumor weight of group 4TRS005 single-drug group was 0.508±0.063g, and the tumor inhibition rate was 76.09%; the tumor weight of group 5TRS005+Gemox group was 0.031±0.019g, and the tumor inhibition rate was 98.54%.

[0169] Table 4. Antitumor activity of TRS005 alone or in combination with Gemox

[0170]

[0171] *: P<0.05 compared with vehicle group

[0172] **: P<0.01 compared with vehicle group

[0173] ***: P<0.001 compared with vehicle group

[0174] Referring to Table 5, the significance analysis of tumor inhibition activity showed that the tumor inhibition rate of group 5 TRS005+Gemox group was significantly higher than that of group 2 R+Gemox group and group 4 TRS005 monotherapy group, and the tumor inhibition rate of group 4 TRS005 monotherapy group was better than that of group 2 R+Gemox group. Therefore, the combination of TRS005 and Gemox of the present invention achieved a more significant tumor inhibition effect than the second-line standard therapy R combined with Gemox, and achieved unexpected technical effects.

[0175] Day 0-13, group 3 received first-line standard therapy R-CHOP, Figure 1 It can be seen that the tumor volume showed a rapid upward trend; on Day 14, after the administration of TRS005 combined with Gemox, the rapid upward trend of the tumor volume was reversed, and the tumor volume began to decrease, indicating that TRS005 combined with Gemox can effectively inhibit the growth of recurrent tumors after R-CHOP treatment.

[0176] Table 5. Analysis of the significance of anti-tumor activity of TRS005 alone or in combination with Gemox

[0177] 2. Effects of the test substances on the body weight of tumor-bearing animals

[0178] The results of the effects of each test substance on the body weight of WSU-DLCL2 tumor-bearing animals showed that on Day 24, the average body weight of each group of animals increased by -1.29%-5.99% (-0.25g-1.3g) compared with the beginning of the experiment on Day 0. The effects of each group of treatments on the body weight of tumor-bearing animals are shown in Table 6.

[0179] Table 6 Effects of drug treatment in each group on the body weight of tumor-bearing animals

[0180]

[0181] During the experiment, the weight changes of mice in each treatment group were within the normal range, no drug-related mouse deaths occurred, and no other obvious drug-related toxic and side effects were observed, indicating that the TRS005 monotherapy and the various TRS005 combination administration regimens of the present invention have good safety.

[0182] Example 4: Evaluation of the anti-tumor efficacy of TRS005 combined with Gemox in the human lymphoma OCI-LY19 model

[0183] (I) Grouping and drug administration

[0184] When the average tumor volume reaches 100-120mm 3 At 14:00, the animals were randomly divided into groups according to the tumor volume, so that the difference in tumor volume between groups was less than 10% of the mean. The day of grouping was designated as Day 0, and drug administration was started according to the animal weight. The grouping and drug administration scheme are shown in Table 7.

[0185] Table 7. Grouping and dosing regimen

[0186]

[0187] Among them, IV means intravenous injection; IP means intraperitoneal injection; QW×3wks means administration once a week for three weeks; BIW×3wks means administration twice a week for three weeks.

[0188] During the experiment, the animal body weight and tumor volume were measured twice a week, and the clinical symptoms of the animals were observed and recorded once a day. 3 After the final weighing, CO 2Euthanasia was performed, and the remaining animals in the same group continued to be administered and observed until the animal welfare endpoint or the experimental endpoint was reached. The experimental period was 28 days (3 weeks of administration and 1 week of observation after drug withdrawal). After the last weighing at the experimental endpoint, CO 2 The remaining animals were euthanized and the experiment was terminated after the tumors were removed and weighed.

[0189] (II) Test results

[0190] 1. Anti-tumor effect of the test substance

[0191] The experimental period was 28 days. On Day 18, the tumor volume of animals in the vehicle control group began to exceed 2000mm 3 When the end point of the experiment was reached, euthanasia was performed. TV The tumor volume data of the vehicle control group is required for indicator calculation, so the data of Day 18 were used for statistical analysis.

[0192] See Table 8 and Figure 3 In the study of the anti-tumor activity of TRS005 combined with Gemox on the human lymphoma OCI-LY19 model, on Day 18, group 2R+Gemox, the average tumor volume was 621.17±72.62mm 3 , %TGI TV The average tumor volume of group 3TRS005 was 897.52±146.95mm 3 , %TGI TV The mean tumor volume of group 4 TRS005+Gemox was 153.24±32.78mm 3 , %TGI TV The rate was 93.76%. R+Gemox is the second-line standard clinical treatment for DLBCL. Compared with R+Gemox, TRS005+Gemox has a significantly improved tumor inhibition effect, achieving unexpected technical results.

[0193] Day 14, the last dose of medication, Figure 3 It can be seen that after Day 1 8, the tumor volume of mice in Group 2R+Gemox increased rapidly. By Day 25, the tumor volume of 9 mice in Group 2 reached the euthanasia standard. Although the tumor volume of mice in Group 4TRS005+Gemox also increased, the growth rate was lower than that of Group 2. By Day 25, no mouse in Group 4 had a tumor volume that reached the euthanasia standard, and only one mouse had a tumor volume that reached the euthanasia standard on Day 28. It can be seen that after Day 18, the difference in tumor volume between the two groups of mice further expanded, further indicating that the tumor inhibition effect of Group 4TRS005+Gemox is better than that of Group 2R+Gemox.

[0194] Table 8. Antitumor activity of TRS005 combined with Gemox

[0195]

[0196] *: P<0.05 compared with vehicle group

[0197] **: P<0.01 compared with vehicle group

[0198] ***: P<0.001 compared with coal dissolution group

[0199] #: P<0.05 compared with R+Gemox group

[0200] ##: P<0.01 compared with R+Gemox group

[0201] ###: P<0.001 compared with R+Gemox group

[0202] 2. Effects of the test substances on the body weight of tumor-bearing animals

[0203] The results of the effects of each test substance on the body weight of OCI-LY19 tumor-bearing animals showed that on Day 18, the average body weight of each group of animals increased by -0.9%-13.07% (-0.20g-2.90g) compared with the beginning of the experiment on Day 0. The effects of each group of treatments on the body weight of tumor-bearing animals are shown in Table 9.

[0204] Table 9. Effects of each group of drug treatment on the body weight of tumor-bearing animals

[0205]

[0206] During the experiment, the weight changes of mice in each treatment group were within the normal range, no drug-related mouse deaths occurred, and no other obvious drug-related toxic and side effects were observed, indicating that the TRS005 combined with Gemox regimen of the present invention has good safety.

[0207] In summary, in order to improve the actual clinical benefits of non-Hodgkin's lymphoma patients, the present invention starts from the actual clinical needs and takes the clinical standard treatment regimen as a control. The TRS005 combined with Gemox regimen used in the present invention significantly improves the tumor inhibition effect in CD20-positive diffuse large B-cell lymphoma compared with the second-line standard therapy R+Gemox. The above results show that the TRS005 combined with Gemox regimen of the present invention shows better tumor inhibition effect and better treatment potential than the existing second-line standard therapy, and has good safety and the potential to better meet clinical treatment needs.

[0208] The above description of the specific embodiments of the present invention does not limit the present invention. Those skilled in the art may make various changes or modifications based on the present invention. As long as they do not depart from the spirit of the present invention, they should all fall within the scope of the claims attached to the present invention.

Claims

1. Use of a combination of an anti-CD20 antibody-drug conjugate and at least one therapeutic agent in the preparation of a medicament for treating diffuse large B-cell lymphoma, characterized in that: The anti-CD20 antibody drug conjugate has the following structure: Among them, mAb indicates recombinant anti-CD20 monoclonal antibody, and the drug-antibody ratio DAR is 4.2±0.5; The at least one therapeutic agent is gemcitabine and oxaliplatin.

2. The use according to claim 1, characterized in that The diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma.

3. The use according to claim 2, characterized in that The diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma after receiving at least one standard treatment regimen.

4. The use according to any one of claims 1 to 3, characterized in that: The recombinant anti-CD20 monoclonal antibody is selected from rituximab or its biosimilars, ofatumumab or its biosimilars, ocrelizumab or its biosimilars, and obinutuzumab or its biosimilars.

5. The use according to claim 4, characterized in that The recombinant anti-CD20 monoclonal antibody is selected from rituximab or its biosimilars.

6. The use according to claim 5, characterized in that The diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

7. The use according to claim 6, characterized in that The CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma that double-expresses MYC and BCL2.

8. The use according to any one of claims 5 to 7, characterized in that: The at least one standard treatment regimen includes at least anti-CD20 antibody monotherapy or anti-CD20 antibody combination therapy.

9. The use according to claim 8, characterized in that The at least one standard treatment regimen includes at least one selected from R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R and Pola-R-CHP.

10. The use according to claim 9, characterized in that The at least one standard treatment regimen is R-CHOP.

11. A drug combination for treating diffuse large B-cell lymphoma, characterized in that: The drug combination is a combination of an anti-CD20 antibody-drug conjugate, gemcitabine and oxaliplatin, and the anti-CD20 antibody-drug conjugate has the following structure: Among them, mAb represents recombinant anti-CD20 monoclonal antibody, and the drug-antibody ratio DAR is 4.2±0.

5.

12. The pharmaceutical combination according to claim 11, characterized in that The diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma.

13. The pharmaceutical combination according to claim 12, characterized in that The diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma after receiving at least one standard treatment regimen.

14. The pharmaceutical combination according to any one of claims 11 to 13, characterized in that The recombinant anti-CD20 monoclonal antibody is selected from rituximab or its biosimilars, ofatumumab or its biosimilars, ocrelizumab or its biosimilars, and obinutuzumab or its biosimilars.

15. The pharmaceutical combination according to claim 14, characterized in that The recombinant anti-CD20 monoclonal antibody is selected from rituximab or its biosimilars.

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