Pharmaceutical composition for treating non-Hodgkin's lymphoma and its use

Abstract

The present invention discloses the use of an anti-CD20 antibody-drug conjugate or a pharmaceutical composition thereof in the manufacture of a therapeutic agent for non-Hodgkin's lymphoma, the pharmaceutical composition comprising the anti-CD20 antibody-drug conjugate and at least one therapeutic agent. Furthermore, the combination therapy with the anti-CD20 antibody-drug conjugate or a pharmaceutical composition comprising the same has a superior therapeutic effect to existing clinical second-line standard therapies.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This patent application claims priority from Chinese invention patent application bearing application number CN 202311225386.1, filed on September 21, 2023, the entire contents of which are incorporated herein by reference.

[0002] Technical Field The present invention relates to the field of pharmaceuticals and drug combinations, and in particular to pharmaceutical compositions and uses thereof for the treatment of non-Hodgkin's lymphoma. [Background technology]

[0003] Non-Hodgkin lymphoma (NHL) is the seventh most common cancer in the world, accounting for approximately 4% to 5% of new cases and 3% to 4% of deaths. Worldwide, the disease affects approximately 1.5 million people.

[0004] NHL is a general term for malignant proliferative disorders of the lymphatic system, including various types of lymphoma. These tumors are primarily derived from B lymphocytes (>85%), with a small proportion derived from T cells and natural killer cells. NHL is usually classified as indolent or indolent NHL (iNHL) or aggressive or aggressive NHL (aNHL) based on prognosis. Follicular lymphoma (FL) is the most common type of iNHL, accounting for approximately one-fifth of all NHL cases, while diffuse large B-cell lymphoma (DLBCL) is the most common type of aNHL, accounting for approximately one-third of all NHL cases.

[0005] Thirty to 35% of DLBCLs express the MYC protein, and 20 to 35% co-express BCL2, which are termed "dual-expressing lymphomas" and suggest a poor prognosis.

[0006] Improving disease treatment efficacy through drug combinations is a common concept in this field. However, in clinical practice, the standard drug combination treatment regimens adopted for different treatment lines and different subtypes of non-Hodgkin's lymphoma vary, and even more drug combination regimens are under research. The types of chemical and biological drugs that can be used in combination are even more complex. This presents great difficulties in designing drug combination regimens, and there is great uncertainty regarding the therapeutic efficacy and safety of drug combinations.

[0007] WO2021223048A1 and its Chinese equivalent CN115485304A disclose anti-CD20 antibody-drug conjugates, and the specifications state that the conjugates can be used in combination with doxorubicin, cyclophosphamide, taxanes, capecitabine, gemcitabine, irinotecan, oxaliplatin, prednisone, vincristine, etc., or combinations thereof. The applicant's prior applications CN108452318A and CN108452319A each disclose anti-CD20 antibody-drug conjugates and formulations thereof, and the instructions state that the anti-CD20 antibody-drug conjugates can be used in combination with cyclophosphamide, doxorubicin, vincristine, prednisone, PD1 antibodies, CTLA4 inhibitors, or combinations thereof.

[0008] However, the above-mentioned patent application merely lists the combination of anti-CD20 conjugates with commonly used anti-tumor drugs in clinical practice, and the examples do not include specific drug combination regimens for the treatment of non-Hodgkin's lymphoma, nor does it provide data on therapeutic efficacy or safety. Furthermore, drugs such as taxanes, PD1 antibodies, and CTLA4 inhibitors used in combination with the aforementioned anti-CD20 antibody-drug conjugates have not even been used clinically to treat non-Hodgkin's lymphoma. Therefore, listing the various drugs used in the above-mentioned patents is merely a method of patent preparation and constitutes accidental disclosure. Due to the large number of drug types and combination regimens that can be used in combination, and the uncertainty surrounding the therapeutic efficacy and toxicity of the combined drugs, the above-mentioned patent application fails to provide technical revelation regarding specific drug combination regimens.

[0009] Currently, the first-line standard treatment regimens for DLBCL recommended by the 2022 edition of the Clinical Practice Guidelines for Lymphoma issued by the Chinese Society of Clinical Oncology (CSCO) are shown in Table a below. These regimens include R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R, and Pola-R-CHP. R-CHOP is a combination of rituximab (R), cyclophosphamide (C), vincristine (H), doxorubicin (O), and prednisone (P). R-CHOEP is a combination of rituximab (R), cyclophosphamide (C), vincristine (H), The R-miniCHOP regimen is a combination of rituximab (R), cyclophosphamide (C), vincristine (H), doxorubicin (O), and prednisone (P), in which CHOP is used at a low dose; the DA-EPOCH-R regimen is a combination of etoposide (E), prednisone (P), doxorubicin (O), cyclophosphamide (C), vincristine (H), and rituximab (R), with dose adjustments after each chemotherapy cycle according to the patient's response to chemotherapy. Adjustment, DA); Pola-R-CHP is a combination of Polatuzumab vedotin (an ADC targeting CD79b, trade name Polivy®), rituximab (R), cyclophosphamide (C), vincristine (H) and prednisone (P).

[0010] [Table 1]

[0011] Even after undergoing the above-mentioned first-line treatments, patients achieve varying degrees of relief, but 40% to 50% of cases are still prone to relapse, are refractory, and have a poor prognosis, especially for "double-expression lymphoma."

[0012] Currently, the second-line standard treatment regimens for relapsed or refractory DLBCL after treatment with the first-line standard treatment regimen recommended in the 2022 edition of the Clinical Practice Guidelines for Lymphoma issued by the Chinese Society of Clinical Oncology (CSCO) are shown in Table b below. These regimens include R-GemOx (combination of rituximab (R), gemcitabine (Gem), and oxaliplatin (Ox)), R2 (combination of rituximab (R) and lenalidomide (R)), BR (combination of bendamustine (B) and rituximab (R)), and R-ICE (combination of rituximab, ibo). These include 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), and DA-EPOCH-R (combination of rituximab, etoposide, prednisone, vincristine, cyclophosphamide, and doxorubicin). Various of the above-mentioned regimens recommended in the guidelines, such as the R-GemOx regimen, have also become standard treatments for other subtypes of non-Hodgkin's lymphoma, such as mantle cell lymphoma and marginal zone lymphoma.

[0013] [Table 2]

[0014] Although patients can achieve a certain degree of relief after undergoing the above-mentioned second-line standard treatment regimens, there is still room for further improvement in clinical benefits. Therefore, it is necessary to explore more effective treatments, combination drugs, and treatment regimens for non-Hodgkin's lymphoma than the existing standard therapies. Summary of the Invention [Problem to be solved by the invention]

[0015] The object of the present invention is to provide a pharmaceutical composition and its use in the manufacture of a therapeutic agent for non-Hodgkin's lymphoma, and the combination therapy of the pharmaceutical composition has a superior therapeutic effect compared with the existing clinical second-line standard therapy. [Means for solving the problem]

[0016] A first aspect of the present invention discloses the use of an anti-CD20 antibody-drug conjugate in the manufacture of a medicament for the treatment of non-Hodgkin's lymphoma.

[0017] The anti-CD20 antibody-drug conjugate has the following structural formula:

[0018] mAb-(LD)n;

[0019] In the formula, mAb is a recombinant anti-CD20 monoclonal antibody, D is a small molecule toxin, L is a linker connecting the monoclonal antibody and the small molecule toxin, n is the average number of small molecule toxins bound to the monoclonal antibody, and "-" represents a bond.

[0020] Preferred recombinant anti-CD20 monoclonal antibodies include, but are not limited to, rituximab or a biologically similar drug thereof, ofatumumab or a biologically similar drug thereof, obinutuzumab or a biologically similar drug thereof, and ocrelizumab or a biologically similar drug thereof.

[0021] More preferably, the recombinant anti-CD20 monoclonal antibody is rituximab or a biologically similar drug thereof.

[0022] Preferably, D is one or more monomethyl auristatins, including, but not limited to, monomethyl auristatin-E (MMAE), monomethyl auristatin-D (MMAD), or monomethyl auristatin-F (MMAF); further, the monomethyl auristatin is monomethyl olestatin-E (MMAE).

[0023] L is a cleavable linker or a non-cleavable linker, including, but not limited to, a thioether-linked linker, which includes a maleimide group. Preferred maleimide groups include a maleimidocaproyl group (mc) that further contains caproic acid. Preferred maleimide groups also include maleimidomethyl groups, such as succinimide-4-(N-maleimidomethyl)cyclohexane-1-carboxylic acid ester (sMCC) or sulfosuccinimide-4-(N-maleimidomethyl)cyclohexane-1-carboxylic acid ester (sulfo-sMCC).

[0024] Cleavable linkers include, but are not limited to, hydrazone bond linkers, disulfide bond linkers, and peptide bond linkers, including valine-citrulline (Val-Cit or VC), phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), or valine-lysine (Val-Lys). A preferred peptide bond linker is valine-citrulline (Val-Cit).

[0025] Additionally, the linker moiety includes para-aminobenzyl (PAB), para-amino-benzyloxycarbonyl (PABC) or derivatives or analogs thereof.

[0026] Furthermore, L is preferably MC-VC-PAB.

[0027] n is an integer or a non-integer of 1 to 8; preferably, n is 4.2±1, more preferably 4.2±0.5, and even more preferably 4.2±0.3.

[0028] The anti-CD20 antibody-drug conjugate further has the structure: [ka]

[0029] In the formula, mAb is rituximab or a biologically similar drug thereof, and the average number n of small molecule toxins bound to the antibody is 1 to 8, preferably n is 4.2±1, more preferably n is 4.2±0.5, and even more preferably n is 4.2±0.3.

[0030] The anti-CD20 antibody-drug conjugate of the present invention may be a pharmaceutical product containing the CD20 antibody-drug conjugate and a carrier or excipient, and the pharmaceutical product may be a liquid or lyophilized preparation.

[0031] The carrier or excipient is selected from the group consisting of a pH buffering agent, an osmolality adjusting agent, a lyophilized powder excipient, a protein protecting agent, a solubilizing agent, and water for injection.

[0032] Examples of the pH buffer include histidine hydrochloride buffer, acetate buffer, phosphate buffer, citrate buffer, and Tris buffer, and a preferred pH buffer is histidine hydrochloride buffer.

[0033] The protein protecting agent includes trehalose, sucrose, lactose, or glucose, and a preferred protein protecting agent is trehalose.

[0034] The osmolality adjuster includes mannitol or sodium chloride, and a preferred osmolality adjuster is mannitol.

[0035] The solubilizing agent may be polysorbate 80, polysorbate 20, or poloxamer 188, and a preferred solubilizing agent is polysorbate 80.

[0036] The liquid or lyophilized formulation further comprises the anti-CD20 antibody-drug conjugate, histidine hydrochloride, trehalose, mannitol, and polysorbate 80.

[0037] Preferably, the anti-CD20 antibody-drug conjugate is used in combination with at least one therapeutic agent, the at least one therapeutic agent being selected from antimetabolites and platinum agents.

[0038] the antimetabolite is one or more selected from the group consisting of pemetrexed, methotrexate, fluorouracil, capecitabine, cytarabine, gemcitabine, 6-mercaptopurine, and thioguanine;

[0039] The platinum agent is one or more selected from the group consisting of carboplatin, cisplatin, and oxaliplatin.

[0040] Additionally, the at least one therapeutic agent is gemcitabine and oxaliplatin.

[0041] Preferably, the non-Hodgkin's lymphoma is relapsed or refractory non-Hodgkin's lymphoma.

[0042] Preferably, the relapsed or refractory non-Hodgkin's lymphoma is a relapsed or refractory non-Hodgkin's lymphoma after receiving at least one standard treatment regimen.

[0043] Preferably, the non-Hodgkin's lymphoma is selected from the group consisting of small lymphocytic lymphoma, B-cell chronic lymphocytic lymphoma, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, mantle cell lymphoma, precursor B-cell lymphoblastic leukemia / lymphoma (B-ALL / LBL), T-cell large granular lymphocytic leukemia or anaplastic large cell lymphoma (ALCL).

[0044] Preferably, the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.

[0045] Preferably, the diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma.

[0046] Furthermore, the diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma after receiving at least one standard treatment regimen.

[0047] Furthermore, the diffuse large B-cell lymphoma is a CD20-positive diffuse large B-cell lymphoma.

[0048] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with dual expression of MYC and BCL2.

[0049] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with high CD20 expression or low CD20 expression.

[0050] 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. Still further, 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.

[0051] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma in which CD20 is expressed at a low level and one or more of MYC, BCL2, and BCL6 are expressed at a high level.

[0052] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with low CD20 expression and high MYC and BCL2 expression.

[0053] Preferably, the at least one standard treatment regimen includes at least an anti-CD20 antibody monotherapy or anti-CD20 antibody combination therapy.

[0054] Furthermore, the at least one standard treatment regimen comprises at least one selected from the group consisting of R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R, and Pola-R-CHP.

[0055] Still further, the at least one standard treatment regimen is R-CHOP.

[0056] A second aspect of the present invention discloses a pharmaceutical composition for treating non-Hodgkin's lymphoma.

[0057] The pharmaceutical composition comprises an anti-CD20 antibody-drug conjugate and at least one therapeutic agent, wherein the anti-CD20 antibody-drug conjugate has the following structure: [ka] ;

[0058] Here, the mAb is a recombinant anti-CD20 monoclonal antibody, and the drug-antibody ratio (DAR) is 1 to 8, with the preferred DAR being 4.2±1, the more preferred DAR being 4.2±0.5, and the even more preferred DAR being 4.2±0.3.

[0059] Preferred recombinant anti-CD20 monoclonal antibodies include rituximab or biologically similar drugs thereof, ofatumumab or biologically similar drugs thereof, ocrelizumab or biologically similar drugs thereof, and obinutuzumab or biologically similar drugs thereof.

[0060] More preferably, the recombinant anti-CD20 monoclonal antibody is selected from the group consisting of Rituximab or its biological analogs.

[0061] Preferably, the at least one therapeutic agent is selected from an antimetabolite and a platinum agent, the antimetabolite being one or more selected from the group consisting of pemetrexed, methotrexate, fluorouracil, capecitabine, cytarabine, gemcitabine, 6-mercaptopurine, and thioguanine, and the platinum agent being one or more selected from the group consisting of carboplatin, cisplatin, and oxaliplatin.

[0062] Additionally, the at least one therapeutic agent is gemcitabine and oxaliplatin.

[0063] Preferably, the pharmaceutical composition of the present invention is a combination of the anti-CD20 antibody-drug conjugate, gemcitabine, and oxaliplatin.

[0064] The above-mentioned combinations of anti-CD20 antibody-drug conjugates and at least one therapeutic agent are merely exemplary combinations and are not intended to limit the combinations of the anti-CD20 antibody-drug conjugates of the present invention with other drugs.

[0065] The pharmaceutical compositions of the present invention comprise a first pharmaceutical composition comprising the anti-CD20 antibody-drug conjugate and a carrier or excipient therefor, and a second pharmaceutical composition comprising at least one therapeutic agent and a carrier or excipient therefor.

[0066] The first pharmaceutical composition and the second pharmaceutical composition according to the present invention may be packaged independently or may be packaged together.

[0067] The present invention also discloses a kit comprising an anti-CD20 antibody-drug conjugate and at least one therapeutic agent.

[0068] Preferably, the anti-CD20 antibody-drug conjugate and at least one therapeutic agent are separately packaged and disposed within the kit.

[0069] Preferably, the kit comprises the anti-CD20 antibody-drug conjugate, gemcitabine, and oxaliplatin.

[0070] A third aspect of the present invention discloses a combination therapy for treating non-Hodgkin's lymphoma.

[0071] The combination therapy of the present invention comprises administering an effective amount of the pharmaceutical composition according to the second aspect of the present invention for the treatment of non-Hodgkin's lymphoma.

[0072] Preferably, the combination therapy comprises combining effective amounts of the anti-CD20 antibody-drug conjugate, gemcitabine and oxaliplatin.

[0073] Preferably, the non-Hodgkin's lymphoma is relapsed or refractory non-Hodgkin's lymphoma.

[0074] Preferably, the relapsed or refractory non-Hodgkin's lymphoma is a relapsed or refractory non-Hodgkin's lymphoma after receiving at least one standard treatment regimen.

[0075] Preferably, the non-Hodgkin's lymphoma is selected from the group consisting of small lymphocytic lymphoma, B-cell chronic lymphocytic lymphoma, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, mantle cell lymphoma, precursor B-cell lymphoblastic leukemia / lymphoma (B-ALL / LBL), T-cell large granular lymphocytic leukemia or anaplastic large cell lymphoma (ALCL).

[0076] Preferably, the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.

[0077] Preferably, the diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma.

[0078] Furthermore, the relapsed or refractory diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma after receiving at least one standard treatment regimen.

[0079] Furthermore, the diffuse large B-cell lymphoma is a CD20-positive diffuse large B-cell lymphoma.

[0080] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with dual expression of MYC and BCL2.

[0081] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with high CD20 expression or low CD20 expression.

[0082] Furthermore, the CD20-positive diffuse large B-cell lymphoma is diffuse large B-cell lymphoma in which CD20 is highly expressed and one or more of MYC, BCL2, and BCL6 are highly expressed.

[0083] 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.

[0084] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma in which CD20 is expressed at a low level and one or more of MYC, BCL2, and BCL6 are expressed at a high level.

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

[0086] Preferably, the at least one standard treatment regimen includes at least an anti-CD20 antibody monotherapy or anti-CD20 antibody combination therapy.

[0087] Furthermore, the at least one standard treatment regimen comprises at least one selected from the group consisting of R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R, and Pola-R-CHP.

[0088] Still further, the at least one standard treatment regimen is R-CHOP.

[0089] The anti-CD20 antibody-drug conjugate is administered before, simultaneously with, and / or after the administration of the at least one therapeutic agent.

[0090] The anti-CD20 antibody-drug conjugate and the at least one therapeutic agent may be administered once or multiple times. For example, the anti-CD20 antibody-drug conjugate may be administered once or multiple times. When administered multiple times, the administration frequency may 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. Alternatively, one cycle may be one day, one week, two weeks, three weeks, four weeks, one month, two months, three months, or six months, with one, two, three, four, five, six, or other administrations being administered in each cycle. Administration treatment is continued at an appropriate administration cycle and frequency until the subject is cured, the condition progresses, or the subject dies.

[0091] At least one therapeutic agent, such as gemcitabine or oxaliplatin, may be administered once or multiple times. In the case of multiple administration, the administration frequency may be, for example, three times a day, twice a day, once a day, once every other day, once a week, once every two weeks, once every three weeks, or once every four weeks. Alternatively, one cycle may be one day, one week, two weeks, three weeks, four weeks, one month, two months, three months, or six months, with one, two, three, four, five, six, or other doses administered in each cycle. Administration treatment is continued at an appropriate administration cycle and frequency until the subject is cured, the condition progresses, or the subject dies.

[0092] With regard to dosage, the initial dose candidate of the anti-CD20 antibody-drug conjugate to be administered to a patient may be about 1 μg / kg to 100 mg / kg depending on the type and severity of the disease, and an exemplary dose range of the anti-CD20 antibody-drug conjugate may be, for example, 0.01 mg / kg to 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, with preferred dosages being 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.

[0093] The dosage of at least one therapeutic agent administered to a patient ranges from 1 mg / m to 1 mg / m depending on the type and severity of the disease. 2 ~100g / m 2 Or it may be 1 μg / kg-100 mg / kg. For example, the dose of gemcitabine may be 10 mg / m 2 ~100g / m 2 and preferably 100 mg / m 2 ~10g / m 2 , preferably 200 mg / m 2 ~5000mg / m 2 and the preferred dosage is 250 mg / m 2 , 500 mg / m 2 , 750 mg / m 2 , 1000 mg / m2 , 1500 mg / m 2 , 2000 mg / m 2 , 2500 mg / m 2 For example, the dose of oxaliplatin may be 0.1 mg / m 2 ~10g / m 2 or 0.1 μg / kg-10 mg / kg, preferably the dosage is 1 mg / m 2 ~10g / m 2 may be used, preferably 10 mg / m 2 ~1000mg / m 2 and preferably 20 mg / m 2 ~500mg / m 2 and preferably 25 mg / m 2 , 50 mg / m 2 , 75 mg / m 2 , 100 mg / m 2 , 150 mg / m 2 , 200 mg / m 2 , 250 mg / m 2 etc. may also be used.

[0094] The combination drug of the present invention can be administered by any suitable means, including gastrointestinal, parenteral, pulmonary, intranasal, etc. Parenteral administration includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration, etc.

[0095] A fourth aspect of the present invention discloses a method for inhibiting the growth of non-Hodgkin's lymphoma cells.

[0096] The growth-inhibiting method comprises adding to the non-Hodgkin's lymphoma cell line an effective amount of an anti-CD20 antibody-drug conjugate according to the present invention or a pharmaceutical composition according to the second aspect of the present invention.

[0097] The first method of growth inhibition comprises adding to the non-Hodgkin's lymphoma cell line an effective amount of an anti-CD20 antibody-drug conjugate, the anti-CD20 antibody-drug conjugate having the following structure: [ka] ;

[0098] Here, the mAb is a recombinant anti-CD20 monoclonal antibody, and the drug-antibody ratio (DAR) is 1 to 8, with the preferred DAR being 4.2±1, the more preferred DAR being 4.2±0.5, and the even more preferred DAR being 4.2±0.3.

[0099] Preferably, the recombinant anti-CD20 monoclonal antibody is selected from the group consisting of Rituximab or its biosimilars.

[0100] Preferably, the DAR is 4.2±0.5.

[0101] The second method of growth inhibition comprises administering to the non-Hodgkin's lymphoma cell line an effective amount of a pharmaceutical agent according to the second aspect of the invention.

[0102] Preferably, the growth inhibiting method comprises administering to the non-Hodgkin's lymphoma cell line effective amounts of the anti-CD20 antibody-drug conjugate, gemcitabine, and oxaliplatin.

[0103] Preferably, the non-Hodgkin's lymphoma is relapsed or refractory non-Hodgkin's lymphoma.

[0104] Preferably, the relapsed or refractory non-Hodgkin's lymphoma is a relapsed or refractory non-Hodgkin's lymphoma after receiving at least one standard treatment regimen.

[0105] Preferably, the non-Hodgkin's lymphoma is selected from the group consisting of small lymphocytic lymphoma, B-cell chronic lymphocytic lymphoma, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, mantle cell lymphoma, precursor B-cell lymphoblastic leukemia / lymphoma (B-ALL / LBL), T-cell large granular lymphocytic leukemia or anaplastic large cell lymphoma (ALCL).

[0106] Preferably, the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.

[0107] Preferably, the diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma.

[0108] Furthermore, the diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma after receiving at least one standard treatment regimen.

[0109] Preferably, the diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

[0110] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with dual expression of MYC and BCL2.

[0111] Preferably, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with high CD20 expression or low CD20 expression.

[0112] Preferably, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma in which CD20 is highly expressed and one or more of MYC, BCL2, and BCL6 are highly expressed.

[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 MYC and BCL2.

[0114] Preferably, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma in which CD20 is expressed at a low level and one or more of MYC, BCL2, and BCL6 are expressed at a high level.

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

[0116] Preferably, the at least one inhibitory treatment comprises an anti-CD20 antibody alone or in combination with at least one therapeutic agent.

[0117] Additionally, the at least one inhibitory treatment comprises at least one selected from the group consisting of R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R, and Pola-R-CHP.

[0118] Additionally, the at least one inhibitory treatment is R-CHOP.

[0119] Based on actual clinical needs and using clinical standard treatment regimens as a control, the present invention demonstrates that the combination regimen of the anti-CD20 antibody-drug conjugate of the present invention with Gemox for non-Hodgkin's lymphoma, particularly CD20-positive diffuse large B-cell lymphoma, exhibits superior tumor-inhibitory effects and therapeutic potential compared to existing second-line standard treatment regimens, while at the same time having good safety profile.

[0120] It should be understood that one, some, or all of the various embodiments described herein may be combined to form other embodiments of the present invention. These and other embodiments of the present invention are further described in the detailed description that follows.

[0121] Hereinafter, an embodiment of the present invention will be described in detail with reference to the accompanying drawings. [Brief explanation of the drawings]

[0122] [Figure 1] : Changes in tumor volume in mice in each treatment group in the WSU-DLCL2 model; [Figure 2] : Tumor weight of mice in each treatment group at the end of the study in the WSU-DLCL2 model; [Figure 3] : Trends in tumor volume change in mice for each treatment group in the OCI-LY19 model. DETAILED DESCRIPTION OF THE INVENTION

[0123] Specific Embodiments

[0124] The present invention will now be described with reference to specific examples, which should be understood by those skilled in the art as being illustrative of the present invention and not limiting the scope of the present invention in any way.

[0125] The experimental methods in the following examples are conventional methods unless otherwise specified. Raw materials, reagents, etc. used in the following examples are commercially available products unless otherwise specified.

[0126] Definition:

[0127] "Rituximab" refers to a monoclonal antibody drug targeting CD20, developed by Roche and approved by the National Medical Products Regulatory Authority and marketed under the trade name "Mei Luohua." Rituximab is its generic name; only the original drug can be called rituximab.

[0128] "Biologically similar drugs to rituximab" refers to monoclonal antibody drugs, excluding the original drug rituximab, that have been developed by other companies to imitate rituximab, whose amino acid sequence is identical to that of rituximab and whose physicochemical properties, therapeutic effects, pharmacokinetics and safety are similar to those of the original drug rituximab.

[0129] "High expression" means that a technician recognizes high expression of an antigen or gene when the expression level of the antigen or gene in tumor tissue or cells exceeds a certain expression level or a relatively high expression level based on conventional evaluation criteria, for example, strong positivity (+++) and / or positivity (++) determined based on the proportion of immunohistochemically stained cells and staining intensity.

[0130] "Low expression" means that a technician determines that an antigen or gene has low expression when the expression level of the antigen or gene in tumor tissue or cells is below a certain expression level or a relatively low expression level based on conventional evaluation criteria, for example, based on the proportion of immunohistochemically stained cells and staining intensity as weakly positive (+).

[0131] An "effective dose" refers to an amount effective, at a desired dosage and for a desired period of time, to achieve a desired therapeutic result. An effective dose can vary depending on 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.

[0132] Having generally described the present invention, the following examples further disclose embodiments of the invention and are not to be construed as limiting the scope of the claims.

[0133] Example 1: Preparation of anti-CD20 ADC (TRS005) For an exemplary method for producing anti-CD20 ADC (TRS005), see Chinese Patent Application CN108452318B, the entire text of which is incorporated herein by reference.

[0134] The structure of TRS005 is as follows: [ka] ;

[0135] Here, the mAb is rituximab or a biosimilar of rituximab with a drug-antibody ratio (DAR) of 4.2±0.5.

[0136] Example 2: Screening of human lymphoma cell lines The expression levels (mRNA levels) of BCL2, MYC, and CD20 in the lymphoma cell lines SU-DHL-4, SU-DHL-6, TMD-8, WSU DLCL2, DOHH2, DB, OCI-LY10, and OCI-LY19 were measured by sequencing. Two lymphoma cell lines were selected: WSU-DLCL2, which expresses both BCL2 and MYC, but has high CD20 expression, and OCI-LY19, which expresses both BCL2 and MYC and has low CD20 expression. WSU-DLCL2 and OCI-LY19 are CD20-positive and have dual expression of both BCL2 and MYC. The expression levels of CD20, BCL2, and MYC in both cell lines are shown in Table 1.

[0137] [Table 3]

[0138] Example 3: Evaluation of antitumor effects of TRS005 alone or in combination with Gemox in a human lymphoma WSU-DLCL2 / R-CHOP relapse model

[0139] (1) Establishing a model The recurrence model was established using standard methods in the art. For example, WSU-DLCL2 cells were inoculated into mice and treated with R-CHOP (P1). Tumor tissues with good growth from WSU-DLCL2 / R-CHOP (P1) were selected, and the mouse fascia and internal necrotic tissue were removed. The resulting block was inoculated into mice under aseptic conditions and treated with R-CHOP (P2). Tumor tissues with good growth from WSU-DLCL2 / R-CHOP (P2) were selected, and the mouse fascia and internal necrotic tissue were removed. The resulting block was inoculated into mice under aseptic conditions and treated with R-CHOP (P3). The R-CHOP recurrence model was established through P1-P3 subculture.

[0140] WSU-DLCL2 / R-CHOP (P3) tumor tissue showing good growth was selected, and after removing mouse fascia and internal necrotic tissue, it was cut into 2 x 2 x 2 mm blocks and stored in RPMI 1640 basal medium. After isoflurane anesthesia, the tissue block was inoculated subcutaneously on the right side of the mouse using an 11G trocar under sterile conditions.

[0141] (2) Grouping and Administration The average tumor volume was approximately 150 mm 3 When tumor volume reached 1000 mg / kg, the animals were randomly divided according to tumor volume so that the difference in tumor volume between groups was less than 10% of the mean. In this experiment, six animals were used per group. The day of grouping was designated Day 0, and animals were immediately dosed according to their weight after grouping. On Day 24, the remaining animals were euthanized, and tumors were collected, weighed, photographed, and recorded. The experiment was then terminated. The grouping and dosing regimen are shown in Tables 2-3 below.

[0142] [Table 4]

[0143] [Table 5]

[0144] On days 14-20, the administration regimen for Group 3 was changed to TRS005-Gemox, while the administration regimen for the other groups remained unchanged on days 14-20.

[0145] During the study, animal weights and tumor volumes were measured twice weekly. Clinical observations were recorded daily, including the animals' overall health, weight and behavioral abnormalities, and any other treatment-related adverse reactions.

[0146] (3) Evaluation index and statistical analysis Tumor growth inhibition rate (%TGI TV ) is (1-TV T / TV C )×100%, TV Cis the mean tumor volume in the negative control group, TV T was calculated based on the mean tumor volume of the treatment group.

[0147] Relative tumor volume (RTV) was calculated as Vt / V0, where V0 is the tumor volume at the time of grouping and Vt is the tumor volume at each measurement.

[0148] Relative tumor growth rate (%T / C RTV ) is T RTV / C RTV The calculation was done by multiplying the T by 100%. RTV is the treatment group RTV, and C RTV is the negative control group RTV.

[0149] Tumor inhibition rate (%TGI TW ) is (TW C -TW T ) / TW C × 100%, where TW C is the mean tumor weight of the negative control group, and TW T is the mean tumor weight of the treatment group.

[0150] The percent weight change (%BWC) of the animals was calculated using the formula (BWt-BW0) / BW0 x 100%, where BWt is the weight of the animals at each measurement, and BW0 is the weight of the animals at the time of grouping.

[0151] According to the China NMPA "Technical Guidelines for Non-clinical Research on Cytotoxic Anticancer Drugs" (November 2006), %T / C RTV A statistical analysis of ≦40% and P<0.05 was considered significant. If the drug-related animal mortality rate exceeded 20%, the drug dose was considered to be severely toxic.

[0152] All experimental data are expressed as mean ± SEM, and comparisons between groups were performed using a two-tailed t-test, with P < 0.05 considered significant and P < 0.01 considered highly significant (Microsoft Excel 2007, Redmond, WA, USA).

[0153] (4) Test results 1. Tumor-inhibitory effect of the test substance In this experiment, the results of investigating the antitumor activity of TRS005 alone or in combination with Gemox against a human lymphoma WSU-DLCL2 / R-CHOP relapse model are shown in Tables 4-5 and Figures 1-2.

[0154] As shown in Table 4 and Figures 1-2, on Day 24, the tumor weight in Group 2 R+Gemox group was 0.705±0.199g, and the tumor inhibition rate was 66.82%; the tumor weight in Group 4 TRS005 single agent group was 0.508±0.063g, and the tumor inhibition rate was 76.09%; and the tumor weight in Group 5 TRS005+Gemox group was 0.031±0.019g, and the tumor inhibition rate was 98.54%. [Table 6]

[0155] Referring to Table 5, the significance analysis of tumor inhibitory activity showed that the tumor inhibition rate of the TRS005 + Gemox group in Group 5 was significantly higher than that of the R + Gemox group in Group 2 and the TRS005 monotherapy group in Group 4, and the tumor inhibition rate of the TRS005 monotherapy group in Group 4 was superior to that of the R + Gemox group in Group 2. Therefore, the TRS005 combined with Gemox of the present invention achieved a more significant tumor inhibition effect than the combination of R and Gemox, which is the second-line standard therapy, and achieved an unexpected technical effect.

[0156] On days 0-13, Group 3 received the first-line standard therapy, R-CHOP. Figure 1 shows that tumor volume showed a tendency to increase relatively rapidly. On day 14, after administration of TRS005 and Gemox in combination, the rapid increase in tumor volume reversed and the tumor volume began to decrease, indicating that the combination of TRS005 and Gemox can effectively suppress the growth of recurrent tumors after R-CHOP treatment.

[0157] [Table 7]

[0158] 2. Effect of the test substance on the body weight of tumor-bearing animals The effect of each test substance on the body weight of WSU-DLCL2 tumor-bearing animals was examined. On Day 24, the average body weight of animals in each group increased by -1.29% and -5.99% (-0.25g-1.3g) compared to the start of the experiment on Day 0. The effect of treatment in each group on the body weight of tumor-bearing animals is shown in Table 6.

[0159] [Table 8]

[0160] During the experimental period, the weight changes of mice in each treatment group were within the normal range, no drug-related deaths occurred, and no other obvious drug-related toxicities or adverse reactions were observed, demonstrating the good safety profile of the TRS005 monotherapy and TRS005 combination therapy regimens of the present invention.

[0161] Example 4: Antitumor efficacy evaluation of TRS005 in combination with Gemox in a human lymphoma OCI-LY19 model

[0162] (1) Grouping and Administration Mean tumor volume 100-120mm 3 When the tumor volume reached 100 mg / kg, the animals were randomly divided according to tumor volume so that the difference in tumor volume between groups was less than 10% of the mean. The day of grouping was designated Day 0, and administration began according to the animal's weight. The grouping and administration regimen are shown in Table 7 below.

[0163] [Table 9]

[0164] During the experiment, the body weight and tumor volume of the animals were measured twice a week, and the clinical symptoms of the animals were observed and recorded once a day. 3After the final weighing, the animals were euthanized with CO2, and the remaining animals in the same group continued to be observed until the animal welfare endpoint or experimental endpoint was reached. The experimental period was 28 days (3 weeks of administration, 1 week of observation after discontinuation). After the final weighing at the experimental endpoint, the remaining animals were euthanized with CO2, and the tumors were collected and weighed, and the experiment was terminated.

[0165] (2) Test results 1. Tumor-inhibitory effect of the test substance The study lasted for 28 days. On Day 18, the tumor volume of the vehicle control animals reached 2000 mm 3 The animals began to exceed the %TGI limit, reaching the end point of the study and euthanasia was performed. TV Because tumor volume data from the vehicle control group is required for index calculation, statistical analysis was performed using data from Day 18.

[0166] As shown in Table 8 and Figure 3, in a study of the antitumor activity of TRS005 in combination with Gemox against the human lymphoma OCI-LY19 model, on Day 18, in Group 2 R+Gemox, the mean tumor volume was 621.17 ± 72.62 mm 3 and %TGI TV The mean tumor volume of TRS005 in Group 3 was 897.52 ± 146.95 mm 3 and %TGI TV In group 4, TRS005+Gemox, the mean tumor volume was 153.24±32.78mm 3 , %TGI TV The efficacy was 93.76%. R+Gemox is the standard second-line treatment for DLBCL, and compared with R+Gemox, the tumor suppression effect of TRS005+Gemox was significantly improved, achieving unexpected technological benefits.

[0167] The last dose was administered on Day 14. As can be seen in Figure 3, after Day 18, tumor volumes in Group 2 R+Gemox mice rapidly increased. By Day 25, tumor volumes in nine Group 2 mice reached the euthanasia criteria; meanwhile, tumor volumes in Group 4 TRS005+Gemox mice also increased, but at a slower rate than Group 2. By Day 25, no Group 4 mice reached the euthanasia criteria, and only one mouse reached the euthanasia criteria on Day 28. After Day 18, the difference in tumor volume between the two groups further widened, further demonstrating the superior tumor-inhibitory effect of Group 4 TRS005+Gemox compared with Group 2 R+Gemox.

[0168] [Table 10]

[0169] 2. Effect of the test substance on the body weight of tumor-bearing animals The effect of each test substance on the body weight of OCI-LY19 tumor-bearing animals was examined. On Day 18, the average body weight of animals in each group increased by -0.9% and -13.07% (-0.20g-2.90g) compared to the start of the experiment on Day 0. The effect of treatment in each group on the body weight of tumor-bearing animals is shown in Table 9.

[0170] [Table 11]

[0171] During the experiment, the weight changes of mice in each treatment group were within the normal range, no drug-related deaths occurred, and no other obvious drug-related toxicities or adverse reactions were observed, demonstrating the good safety profile of the TRS005 and Gemox combination administration regimen of the present invention.

[0172] In light of the above, in order to enhance the actual clinical benefits for patients with non-Hodgkin's lymphoma, the present invention is based on actual clinical needs and uses clinical standard treatment regimens as a control, and has shown that the combined administration regimen of TRS005 and Gemox adopted by the present invention significantly enhances tumor-inhibitory effects compared to the second-line standard treatment, R+Gemox, in non-Hodgkin's lymphoma, particularly CD20-positive diffuse large B-cell lymphoma.The above results show that the combined administration of TRS005 and Gemox of the present invention exhibits superior tumor-inhibitory effects and therapeutic potential compared to existing second-line standard treatment regimens, while at the same time having favorable safety profiles, demonstrating that it can better meet clinical treatment needs.

[0173] The above description of the specific embodiments of the present invention is not intended to limit the present invention, and those skilled in the art may make various modifications or variations based on the present invention without departing from the spirit of the present invention, all of which are within the scope of the claims of the present invention.

Claims

1. 1. Use of a combination of an anti-CD20 antibody-drug conjugate and at least one therapeutic agent in the manufacture of a therapeutic agent for treating non-Hodgkin's lymphoma, wherein the anti-CD20 antibody-drug conjugate has the following structure: 【Chemistry 1】 ； wherein the mAb is a recombinant anti-CD20 monoclonal antibody, the drug-antibody ratio DAR is 4.2±1, preferably the DAR is 4.2±0.5, and the at least one therapeutic agent is gemcitabine and oxaliplatin.

2. 2. The use according to claim 1, wherein the non-Hodgkin's lymphoma is relapsed or refractory non-Hodgkin's lymphoma.

3. The use according to claim 2, wherein the non-Hodgkin's lymphoma is relapsed or refractory non-Hodgkin's 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 the group consisting of rituximab or a biologically similar drug thereof, ofatumumab or a biologically similar drug thereof, ocrelizumab or a biologically similar drug thereof, and obinutuzumab or a biologically similar drug thereof.

5. The use according to claim 4, characterized in that the recombinant anti-CD20 monoclonal antibody is selected from the group consisting of rituximab or its biological analogues.

6. The use according to any one of claims 1 to 5, characterized in that the non-Hodgkin's lymphoma is selected from the group consisting of small lymphocytic lymphoma, B-cell chronic lymphocytic lymphoma, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, precursor B-cell lymphoblastic leukemia / lymphoma (B-ALL / LBL), T-cell large granular lymphocytic leukemia or anaplastic large cell lymphoma (ALCL).

7. 7. The use according to claim 6, wherein the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.

8. The use according to claim 7, wherein the diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

9. The use according to claim 8, wherein the CD20-positive diffuse large B-cell lymphoma is MYC and BCL2 dual-expressing diffuse large B-cell lymphoma.

10. The use according to any one of claims 1 to 9, wherein the at least one standard treatment regimen includes at least anti-CD20 antibody monotherapy or anti-CD20 antibody combination therapy.

11. The use according to claim 10, wherein the at least one standard treatment regimen comprises at least one selected from the group consisting of R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R, and Pola-R-CHP.

12. The use according to claim 11, wherein the at least one standard treatment regimen is R-CHOP.

13. 1. A pharmaceutical composition for use in the treatment of non-Hodgkin's lymphoma, the pharmaceutical composition comprising a combination of an anti-CD20 antibody-drug conjugate, gemcitabine, and oxaliplatin, wherein the anti-CD20 antibody-drug conjugate is characterized by having the following structure: 【Chemistry 2】 ； Here, the mAb is a recombinant anti-CD20 monoclonal antibody, and the drug-antibody ratio DAR is 4.2±1, preferably 4.2±0.

5.

14. The pharmaceutical composition according to claim 13, wherein the non-Hodgkin's lymphoma is relapsed or refractory non-Hodgkin's lymphoma.

15. The pharmaceutical composition according to claim 14, wherein the non-Hodgkin's lymphoma is relapsed or refractory non-Hodgkin's lymphoma after receiving at least one standard treatment regimen.

16. The pharmaceutical composition according to any one of claims 13 to 15, wherein the recombinant anti-CD20 monoclonal antibody is selected from the group consisting of rituximab or a biologically similar drug thereof, ofatumumab or a biologically similar drug thereof, ocrelizumab or a biologically similar drug thereof, and obinutuzumab or a biologically similar drug thereof.

17. 17. The pharmaceutical composition according to claim 16, wherein the recombinant anti-CD20 monoclonal antibody is selected from the group consisting of rituximab or its biologically similar drugs.

18. A combination therapy for treating non-Hodgkin's lymphoma, comprising administering to a subject an effective amount of the pharmaceutical composition according to any one of claims 13 to 17.

19. 19. The combination therapy of claim 18, wherein the non-Hodgkin's lymphoma is selected from the group consisting of small lymphocytic lymphoma, B-cell chronic lymphocytic lymphoma, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, precursor B-cell lymphoblastic leukemia / lymphoma (B-ALL / LBL), T-cell large granular lymphocytic leukemia, or anaplastic large cell lymphoma (ALCL).

20. 20. The combination therapy of claim 19, wherein the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.

21. 21. The combination therapy according to claim 20, wherein the diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

22. 22. The combination therapy according to claim 21, wherein the CD20-positive diffuse large B-cell lymphoma is MYC and BCL2 dual-expressing diffuse large B-cell lymphoma.

23. The combination therapy according to any one of claims 18 to 22, wherein the at least one standard treatment regimen includes at least an anti-CD20 antibody monotherapy or an anti-CD20 antibody combination therapy.

24. The combination therapy of claim 23, wherein the at least one standard treatment regimen comprises at least one selected from the group consisting of R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R, and Pola-R-CHP.

25. The combination therapy of claim 24, wherein the at least one standard treatment regimen is R-CHOP.

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