Pharmaceutical combination for treating non-hodgkin lymphoma and use thereof

Abstract

The use of an anti-CD20 antibody-drug conjugate or a pharmaceutical combination thereof in the preparation of a drug for treating recurrent or refractory non-Hodgkin lymphoma after receiving at least one standard treatment scheme. The pharmaceutical combination comprises an anti-CD20 antibody-drug conjugate and at least one therapeutic agent. Compared with existing clinical second-line standard therapies, the combination therapy using the anti-CD20 antibody-drug conjugate or the pharmaceutical combination comprising same has a better therapeutic effect.

Description

Drug combination for treating non-Hodgkin's lymphoma and use thereof

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This patent application claims the benefit of priority to Chinese invention patent application No. 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 pharmaceuticals and drug combination, and in particular to a drug combination for treating non-Hodgkin's lymphoma and uses 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 disorders of the lymphoid system, encompassing numerous types of lymphocytic neoplasms. These primarily originate from B lymphocytes (>85%), with a smaller proportion arising from T cells and natural killer cells. Based on prognosis, these diseases are typically categorized as slowly progressive or indolent NHL (iNHL) and rapidly progressive or aggressive NHL (aNHL). Follicular lymphoma (FL) is the most common iNHL, accounting for approximately one-fifth of all NHL patients, while diffuse large B-cell lymphoma (DLBCL) is the most common aNHL, accounting for approximately one-third of all NHL patients.

[0006] Among DLBCLs, 30%-35% express MYC protein, and 20-35% also express BCL2, which are called "double-expressing lymphomas." Some DLBCLs simultaneously express MYC, BCL2, and BCL6, which are called "triple-expressing lymphomas," indicating a poor prognosis.

[0007] Improving disease treatment efficacy through drug combination therapy is a common approach in this field. However, the standard drug combination treatment regimens used clinically for different lines and subtypes of non-Hodgkin's lymphoma vary. There are even more drug combination regimens under research, and the variety of chemical and biological drugs that can be used in combination is even more complex. This makes 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 equivalent CN115485304A disclose an anti-CD20 antibody-drug conjugate, and the description mentions 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 the description mentions 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 aforementioned patent application simply lists the combination of anti-CD20 conjugates with commonly used clinical anti-tumor drugs. The examples do not design specific drug combination regimens for the treatment of non-Hodgkin's lymphoma and provide efficacy and safety data. Furthermore, the aforementioned taxanes, PD-1 antibodies, CTLA-4 inhibitors, and other drugs used in combination with anti-CD20 antibody-drug conjugates have not even been used clinically to treat non-Hodgkin's lymphoma. The listing of various drugs for combination use in the aforementioned patent is merely a drafting technique and constitutes incidental disclosure. Due to the wide variety of available combination drugs and regimens, as well as the uncertainty surrounding the efficacy and toxicity of combined drugs, the aforementioned patent application cannot provide technical guidance for specific drug combination regimens.

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

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

[0012] Although the disease progresses, 40%-50% of patients are prone to relapse or refractory disease, especially for "double-expression lymphoma", which shows a poor prognosis.

[0013] The second-line standard treatments for relapsed or refractory DLBCL after first-line standard treatment are 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 (rituximab (R) and lenalidomide (R) combined), BR (bendamustine (B) and rituximab (R) combined), R-ICE (rituximab, ifosfamide, carboplatin and etoposide combined), R-DHAP (rituximab, cisplatin, cytarabine and dexamethasone combined), R-GDP (rituximab, gemcitabine, cisplatin and dexamethasone combined), R-MINE (rituximab, mesna, ifosfamide, mitoxantrone and etoposide combined), R-ESHAP (rituximab, etoposide, methylprednisolone, cisplatin and cytarabine combined), DA-EPOCH-R (rituximab, etoposide, prednisone, vincristine, cyclophosphamide and doxorubicin combined). The above guidelines recommend various regimens such as R 2 The regimen is also used as the standard treatment for other subtypes of non-Hodgkin's lymphoma, such as follicular lymphoma and marginal zone lymphoma.

[0014] Table b Second-line treatment options for DLBCL

[0015] 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 treatments and treatment regimens for relapsed or refractory non-Hodgkin's lymphoma than existing standard treatments.

[0016] Summary of the Invention

[0017] The present invention aims to provide a drug combination and its use in the preparation of a drug for treating non-Hodgkin's lymphoma, wherein the drug combination therapy has a better therapeutic effect than the existing clinical second-line standard therapy.

[0018] The first aspect of the present invention discloses the use of a combination of an anti-CD20 antibody drug conjugate and at least one therapeutic agent in a medicament for treating non-Hodgkin's lymphoma.

[0019] The anti-CD20 antibody drug conjugate has the following structural formula:

[0020] mAb-(LD)n;

[0021] Wherein, mAb represents a recombinant anti-CD20 monoclonal antibody, D represents 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 coupled to the monoclonal antibody; and “-” represents a bond.

[0022] Preferred recombinant anti-CD20 monoclonal antibodies include but are not limited to rituximab or its biosimilars, ofatumumab or its biosimilars, obinutuzumab or its biosimilars, and ocrelizumab or its biosimilars.

[0023] More preferably, the recombinant anti-CD20 monoclonal antibody is rituximab or a biosimilar thereof.

[0024] 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 auristatin-E (MMAE).

[0025] 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, preferably the maleimide group further 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).

[0026] The cleavable linker includes, but is not limited to, a hydrazone linker, a disulfide linker, and a peptide linker; the peptide linker comprises valine-citrulline (Val-Cit or VC), phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), or valine-lysine (Val-Lys). Preferably, the peptide linker is valine-citrulline (Val-Cit).

[0027] Furthermore, the linker portion further comprises p-aminobenzyl alcohol (PAB), p-aminobenzyloxycarbonyl (PABC) or derivatives or analogs thereof.

[0028] Furthermore, L is preferably MC-VC-PAB.

[0029] Said n is an integer or non-integer of 1-8; preferably n is 4.2±1, more preferably 4.2±0.5, and even more preferably 4.2±0.3.

[0030] The anti-CD20 antibody drug conjugate further has the following structure:

[0031] Wherein, mAb is rituximab or a biosimilar thereof, and the average conjugation number n of the small molecule toxin conjugated to the antibody is 1-8; preferably n is 4.2±1, more preferably n is 4.2±0.5, and even more preferably n is 4.2±0.3.

[0032] 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 excipient, and the pharmaceutical preparation is a liquid preparation or a lyophilized preparation.

[0033] The carrier or excipient is selected from the following group: pH buffer, osmotic pressure regulator, lyophilized powder excipient, protein protectant, solubilizer, and water for injection.

[0034] 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.

[0035] The protein protective agent includes trehalose, sucrose, lactose or glucose, and preferably the protein protective agent is trehalose.

[0036] The osmotic pressure regulator includes mannitol or sodium chloride, and the preferred osmotic pressure regulator is mannitol.

[0037] The solubilizer includes polysorbate 80, polysorbate 20 or poloxamer 188, and the preferred solubilizer is polysorbate 80.

[0038] Furthermore, the liquid or lyophilized preparation comprises the above-mentioned anti-CD20 antibody drug conjugate, histidine hydrochloride, trehalose, mannitol and polysorbate 80.

[0039] Preferably, the at least one therapeutic agent is an immunomodulatory drug, and the immunomodulatory drug is selected from one or more of lenalidomide, pomalidomide and thalidomide.

[0040] Furthermore, the at least one therapeutic agent is lenalidomide.

[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 small lymphocytic lymphoma, B-cell chronic lymphocytic leukemia, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, precursor (peripheral) large granular lymphocytic leukemia or anaplastic large cell lymphoma.

[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 CD20-positive diffuse large B-cell lymphoma.

[0048] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma that dually expresses MYC and BCL2.

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

[0051] 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.

[0052] 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.

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

[0054] Furthermore, 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.

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

[0056] The second aspect of the present invention discloses a drug combination for treating non-Hodgkin's lymphoma

[0057] 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:

[0058] Herein, mAb represents a recombinant anti-CD20 monoclonal antibody, and the drug-antibody ratio DAR is 1-8, preferably 4.2±1, more preferably 4.2±0.5, and even more preferably 4.2±0.3.

[0059] Preferably, the recombinant anti-CD20 monoclonal antibody is selected from rituximab or its biosimilar, ofatumumab or its biosimilar, ocrelizumab or its biosimilar, and obinutuzumab or its biosimilar.

[0060] Further preferably, the recombinant anti-CD20 monoclonal antibody is selected from rituximab or a biosimilar thereof.

[0061] Preferably, the at least one therapeutic agent is an immunomodulatory drug, and the immunomodulatory drug is selected from one or more of lenalidomide, pomalidomide and thalidomide.

[0062] Furthermore, the at least one therapeutic agent is lenalidomide.

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

[0064] The above-mentioned combination of the anti-CD20 antibody-drug conjugate and at least one therapeutic agent is merely 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.

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

[0066] The first pharmaceutical composition and the second pharmaceutical composition of the present invention can be packaged independently or co-packaged.

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

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

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

[0070] The 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 drug combination disclosed in the second aspect of the present invention for treating non-Hodgkin's lymphoma.

[0072] Preferably, the combination therapy comprises the combined administration of an effective amount of the anti-CD20 antibody drug conjugate and lenalidomide.

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

[0074] Furthermore, 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 small lymphocytic lymphoma, B-cell chronic lymphocytic leukemia, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, precursor (peripheral) large granular lymphocytic leukemia or anaplastic large cell lymphoma.

[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 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 CD20-positive diffuse large B-cell lymphoma.

[0080] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma that dually expresses MYC and BCL2.

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

[0082] 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. 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.

[0083] 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.

[0084] 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.

[0085] Preferably, the at least one standard treatment regimen includes at least anti-CD20 antibody monotherapy or anti-CD20 antibody combination therapy.

[0086] Furthermore, 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.

[0087] Furthermore, the at least one standard treatment regimen is R-CHOP.

[0088] The anti-CD20 antibody drug conjugate is administered before, simultaneously with, and / or after the at least one therapeutic agent.

[0089] 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 the administration frequency can be one day, one week, two weeks, three weeks, four weeks, one month, two months, three months, or six months as a cycle, with one, two, three, four, five, six, etc., of the drug administered in each cycle, and the administration can be continued according to the appropriate administration cycle and frequency until the subject is cured, the disease symptoms progress, or the subject dies.

[0090] At least one therapeutic agent, such as lenalidomide, can be administered once or multiple times. For multiple administrations, the administration frequency can be, for example, three times a day, twice a day, once a day, every two days, once a week, every two weeks, every three weeks, 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, with administration once, twice, three times, four times, five times, six times, etc. per cycle, and the treatment is continued according to the appropriate dosing cycle and frequency until the subject is cured, disease symptoms progress, or the subject dies.

[0091] 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. An exemplary dosage of the anti-CD20 antibody drug conjugate can range from, 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, 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.

[0092] 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, the dosage of lenalidomide can be 20mg / m 2 -200g / m 2 , preferably 200mg / m 2 -20g / m 2 , preferably 400mg / m 2 -10g / m 2 The preferred dosage is 500 mg / m 2 , 1000mg / m 2 , 1500mg / m 2 , 2000mg / m 2 3000mg / m 2 4000mg / m 2 , 5000mg / m 2 wait.

[0093] 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.

[0094] A fourth aspect of the present invention discloses a method for inhibiting the growth of non-Hodgkin's lymphoma cells.

[0095] 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.

[0096] 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:

[0097] Herein, mAb represents a recombinant anti-CD20 monoclonal antibody, and the drug-antibody ratio DAR is 1-8, preferably 4.2±1, more preferably 4.2±0.5, and even more preferably 4.2±0.3.

[0098] Preferably, the recombinant anti-CD20 monoclonal antibody is selected from rituximab or a biosimilar thereof.

[0099] Preferably, the DAR is 4.2±0.5.

[0100] 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.

[0101] Preferably, the growth inhibition method comprises administering an effective dose of the anti-CD20 antibody drug conjugate and lenalidomide to the non-Hodgkin's lymphoma cell system.

[0102] Preferably, the non-Hodgkin's lymphoma is relapsed or refractory non-Hodgkin's lymphoma.

[0103] Furthermore, 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.

[0104] 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, precursor (peripheral) large granular lymphocytic leukemia or anaplastic large cell lymphoma.

[0105] Preferably, the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.

[0106] Preferably, the diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma.

[0107] Furthermore, the diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma after receiving at least one standard treatment regimen.

[0108] Furthermore, the diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

[0109] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma that dually expresses MYC and BCL2.

[0110] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with high or low CD20 expression.

[0111] 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. 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.

[0112] 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.

[0113] 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.

[0114] 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.

[0115] Furthermore, the at least one inhibitory treatment includes at least one selected from R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R and Pola-R-CHP.

[0116] Furthermore, the at least one inhibitory treatment is R-CHOP.

[0117] Based on actual clinical needs and using clinical standard treatment regimens as a control, the present invention targets non-Hodgkin's lymphoma, especially CD20-positive diffuse large B-cell lymphoma with dual expression of BCL2 and MYC. The anti-CD20 antibody-drug conjugate combination regimen of the present invention shows better tumor inhibition effect and greater therapeutic potential than existing second-line standard treatments, while also having good safety.

[0118] 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

[0119] Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings, in which:

[0120] Figure 1: Changes in tumor volume in mice of each treatment group in the WSU-DLCL2 model;

[0121] Figure 2: Tumor weight of mice in each treatment group at the end of the experiment in the WSU-DLCL2 model;

[0122] Figure 3: Trends in tumor volume changes in mice in each treatment group in the OCI-LY19 model. DETAILED DESCRIPTION

[0123] The present invention is described below with reference to specific examples. Those skilled in the art will appreciate 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.

[0124] 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.

[0125] definition:

[0126] "Rituximab" refers to a CD20-targeting monoclonal antibody developed by Roche and approved by drug regulatory authorities under the trade name "MabThera." Rituximab is its generic name; only brand-name drugs can be called rituximab.

[0127] "Rituximab biosimilars" refer to monoclonal antibody drugs developed by other companies that imitate rituximab, in addition to the original rituximab, with the same amino acid sequence as rituximab and similar physical and chemical properties, efficacy, pharmacokinetics and safety as the original rituximab.

[0128] "High expression" means that technicians identify the expression level of tumor tissue or cell antigens or genes as being higher than a certain expression level or a relatively high expression level according to conventional evaluation standards based on the test results of tumor tissue or cell antigen or gene expression levels. 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 antigens or genes.

[0129] "Low expression" means that technicians identify the expression level of tumor tissue or cell antigens or genes as low expression if it is lower than a certain expression level or a relatively low expression level according to conventional evaluation standards. For example, a weak positive (+) determined by the proportion of immunohistochemically stained cells and the staining intensity can be identified as low expression of antigens or genes.

[0130] An "effective dose" refers to an amount effective to achieve the desired therapeutic result, at dosages and for periods of time necessary. An 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.

[0131]

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

[0132] Example 1: Preparation of anti-CD20 ADC (TRS005)

[0133] The preparation method of an exemplary anti-CD20 ADC (TRS005) is specifically described in Chinese Patent CN108452318B, which is incorporated herein in its entirety.

[0134] The structure of TRS005 is as follows:

[0135] Among them, the mAb was rituximab or a rituximab biosimilar, and the drug-antibody ratio (DAR) was 4.2±0.5.

[0136] Example 2: Human lymphoma cell line screening

[0137] By sequencing, we detected 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. We screened out the lymphoma cell line WSU-DLCL2, which dually expresses BCL2 and MYC and has high CD20 expression, and the cell line OCI-LY19, which dually expresses BCL2 and MYC and has low CD20 expression. WSU-DLCL2 and OCI-LY19 are CD20-positive cell lines that dually express BCL2 and MYC. The expression levels of CD20, BCL2, and MYC in these two cell lines are shown in Table 1.

[0138] Table 1: Expression levels of CD20, BCL2, and MYC in two cell lines (log2TPM)

[0139] Example 3: Evaluation of the antitumor efficacy of TRS005 alone or in combination with lenalidomide in the WSU-DLCL2 / R-CHOP relapse model of human lymphoma

[0140] (1) Model establishment

[0141] The recurrence model was established using conventional methods in the art, such as inoculating WSU-DLCL2 cells into mice and treating them with R-CHOP, which was defined as P1; selecting WSU-DLCL2 / R-CHOP (P1) tumor tissue with good growth status, removing the mouse fascia and internal necrotic tissue, and inoculating mice with plugs under sterile conditions and treating them with R-CHOP, which was defined as P2; selecting WSU-DLCL2 / R-CHOP (P2) tumor tissue with good growth status, removing the mouse fascia and internal necrotic tissue, and inoculating mice with plugs under sterile conditions and treating them with R-CHOP, which was defined as P3; through R-CHOP treatment and passaged from P1 to P3, an R-CHOP recurrence model was established.

[0142] 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.

[0143] (2) Grouping and drug administration

[0144] When the average tumor volume reaches 150 mm 3Around 4:00 p.m., animals were randomly divided into groups based on tumor volume, ensuring that the difference in tumor volume between groups was less than 10% of the mean. Each group consisted of 6 animals. Day 0 was the day of grouping, and dosing was performed immediately after grouping according to animal body weight. On Day 24, the remaining animals were sacrificed, and tumors were weighed, photographed, and recorded. The experiment concluded with the grouping and dosing schedule shown in Tables 2-3.

[0145] Table 2. Dosing regimen for days 0-13

[0146] Among them, IV means intravenous injection, PO means oral administration; QW×2w means once a week for two weeks; BIW×2w means twice a week for two weeks; QD×14 means once a day for 14 times.

[0147] Table 3. Dosage regimen for days 14-20

[0148] On days 14-20, the lenalidomide dose in group 2 was increased to 50 mg / kg, and the dosing regimen for other groups remained unchanged on days 14-20.

[0149] Animal body weight and tumor volume were measured twice weekly during the experiment. Clinical symptoms were observed and recorded daily. Clinical observations included overall health, abnormal weight, abnormal behavior, and other medication-related adverse reactions.

[0150] (3) Evaluation indicators and statistical analysis

[0151] 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.

[0152] The relative tumor volume (RTV) was calculated as: Vt / V0, where V0 was the tumor volume at the time of grouping and Vt was the tumor volume at each measurement.

[0153] Relative tumor proliferation rate (%T / C RTV ) is calculated as follows: RTV / C RTV × 100%, where T RTV RTV for the treatment group, C RTV RTV was the negative control group.

[0154] Tumor inhibition rate (% TGI TW )The calculation formula is: (TW C -TWT ) / TW C 100%, including TW C is the average tumor weight of the negative control group, TW T is the average tumor weight of the treatment group.

[0155] 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.

[0156] According to China NMPA's "Technical Guidelines for Non-clinical Studies of Cytotoxic Anti-tumor Drugs" (November 2006), %T / C RTV If the mortality rate is less than 40% and P<0.05, the drug is considered effective. If the number of drug-related animal deaths exceeds 20%, the drug dose is considered to be severely toxic.

[0157] The experimental data were expressed as Mean ± SEM, and the two-tailed t-test was used for comparison between 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).

[0158] (IV) Test results

[0159] 1. Anti-tumor effect of the test substance

[0160] In this experiment, the anti-tumor activity of TRS005 alone or in combination with lenalidomide in the relapsed human lymphoma WSU-DLCL2 / R-CHOP model is shown in Tables 4-5 and Figures 1-2.

[0161] In the lenalidomide combination regimen, on Days 0–13, the lenalidomide dose in the Group 2R+lenalidomide group was 25 mg / kg; on Days 14–20, the lenalidomide dose in the Group 2R+lenalidomide group was increased to 50 mg / kg, and the lenalidomide dose in the Group 4TRS005+lenalidomide group was maintained at 25 mg / kg.

[0162] See Table 4 and Figures 1-2. On Day 24, the tumor weight of the Group 2R+lenalidomide group was 1.573±0.209 g, and the tumor inhibition rate was 25.98%; the tumor weight of the Group 3TRS005 single-agent group was 0.508±0.063 g, and the tumor inhibition rate was 76.09%; the tumor weight of the Group 4TRS005+lenalidomide group was 0.461±0.115 g, and the tumor inhibition rate was 78.31%.

[0163] Table 4. Antitumor activity of TRS005 alone or in combination with lenalidomide

[0164] *: P<0.05 compared with the vehicle group

[0165] **: P < 0.01 compared with the vehicle group

[0166] ***: P < 0.001 compared with vehicle group

[0167] Referring to Table 5, the significance analysis of tumor inhibition activity showed that the tumor inhibition rate of group 4 TRS005 + lenalidomide group was significantly improved compared with group 2 R + lenalidomide (50 mg / kg) (P < 0.001), that is, at a relatively low dose of lenalidomide, the tumor inhibition rate of group 4 TRS005 + lenalidomide group was still significantly improved compared with group 2 R + lenalidomide group. Therefore, the combination regimen of TRS005 + lenalidomide of the present invention can significantly improve the tumor inhibition rate while reducing the dosage of lenalidomide, and has the potential to further improve the clinical benefit of patients. In addition, the tumor inhibition rate of group 3 TRS005 monotherapy group of the present invention was significantly improved compared with group 2 R + lenalidomide group, indicating that the TRS005 monotherapy of the present invention has achieved a significant improvement in efficacy compared with the second-line standard therapy R + lenalidomide.

[0168] Table 5. Analysis of the significance of antitumor activity of TRS005 alone or in combination with lenalidomide

[0169] 2. Effects of the test substances on the body weight of tumor-bearing animals

[0170] 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 2.09%-6.41% (0.45g-1.41g) compared to the start of the experiment on Day 0. The effects of each treatment on the body weight of tumor-bearing animals are shown in Table 6.

[0171] Table 6 Effects of drug treatment in each group on the body weight of tumor-bearing animals

[0172] 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.

[0173] Example 4: Evaluation of the anti-tumor efficacy of TRS005 alone or in combination with lenalidomide in the human lymphoma OCI-LY19 model

[0174] (1) Grouping and drug administration

[0175] When the average tumor volume reaches 100-120 mm 3At 4 hr, animals were randomly divided into groups based on tumor volume, ensuring that the difference in tumor volume between groups was less than 10% of the mean. Grouping was designated Day 0, and dosing began according to animal body weight. The grouping and dosing schedule are shown in Table 7.

[0176] Table 7 Grouping and dosing regimen

[0177] Among them, IV indicates intravenous injection; PO indicates oral administration; QW×3wks indicates administration once a week, administration for three weeks: BIW×3wks indicates administration twice a week, administration for three weeks: QW×3wks indicates administration once a week, administration for three weeks: QD×21 indicates administration once a day for a total of 21 times.

[0178] 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, animals were euthanized by CO2. The remaining animals in the same group continued to receive medication and observation until reaching the animal welfare endpoint or the experimental end point. The experimental period was 28 days (3 weeks of medication and 1 week of observation after drug withdrawal). After the final weighing at the experimental end point, the remaining animals were euthanized by CO2, and the tumors were removed and weighed before the experiment ended.

[0179] (2) Test results

[0180] 1. Anti-tumor effect of the test substance

[0181] The experimental period was 28 days. On Day 18, the animals in the vehicle control group began to develop tumors with a volume greater than 2000 mm 3 The mice that reached the end point of the experiment were euthanized. TV The tumor volume data of the vehicle control group is required for indicator calculation, so the data on Day 18 are used for statistical analysis.

[0182] See Table 8 and Figure 3 for the study of the anti-tumor activity of TRS005 combined with lenalidomide in the human lymphoma OCI-LY19 model. On Day 18, Group 2 R+lenalidomide, the average tumor volume was 2358.92±199.07mm 3 , %TGI TV The mean tumor volume of Group 3 TRS005+lenalidomide was 808.24±140.98mm 3 , %TGI TV R+Lenalidomide is the standard second-line treatment for DLBCL. Significance analysis showed that TRS005+Lenalidomide significantly improved tumor suppression compared to R+Lenalidomide, achieving unexpected technical results.

[0183] The final dose was administered on Day 14. As shown in Figure 3, after Day 18, tumor volume in mice in Group 2 (R + lenalidomide) rapidly increased, and on Day 21, all mice reached the euthanasia criteria. While tumor volume in mice in Group 3 (TRS005 + lenalidomide) also increased, the rate of increase was slower than in Group 2, and only one mouse reached the euthanasia criteria on Day 21. These results further demonstrate that TRS005 + lenalidomide in Group 4 exhibits superior tumor inhibition efficacy compared to R + lenalidomide in Group 2.

[0184] It should be noted that the OCI-LY19 model is a model with low CD20 expression and is more sensitive to chemotherapy. Although R+lenalidomide showed a mild tumor suppression effect in this animal model, it is a standard treatment option in clinical practice. TRS005 is an ADC containing the small molecule drug MMAE. In animal models with low CD20 expression, TRS005+lenalidomide achieved a significant tumor suppression effect compared to R+lenalidomide, demonstrating greater clinical therapeutic potential.

[0185] Table 8. Antitumor activity of TRS005 combined with lenalidomide

[0186] *: P<0.05 compared with the vehicle group

[0187] **: P < 0.01 compared with the vehicle group

[0188] ***: P < 0.001 compared with vehicle group

[0189] #: P<0.05 compared with R+lenalidomide group

[0190] ##: P<0.01 compared with R+lenalidomide group

[0191] ###: P<0.001 compared with R+lenalidomide group

[0192] 2. Effects of the test substances on the body weight of tumor-bearing animals

[0193] 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 animals in each group increased by 2.14%-13.03% (0.48g-2.94g) compared to the start of the experiment on Day 0. The effects of each treatment on the body weight of tumor-bearing animals are shown in Table 9.

[0194] Table 9. Effects of drug treatment in each group on the body weight of tumor-bearing animals

[0195] 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 lenalidomide regimen of the present invention has good safety.

[0196] In summary, in order to improve the actual clinical benefits of patients with non-Hodgkin's lymphoma, the present invention starts from the actual clinical needs and uses the clinical standard treatment regimen as a control. The TRS005 combined with lenalidomide regimen adopted in the present invention significantly improves the tumor inhibition effect in non-Hodgkin's lymphoma, especially CD20-positive DLBCL, compared with the second-line standard treatment R+lenalidomide, showing better therapeutic potential, while having good safety and can better meet clinical treatment needs.

[0197] The above description of the specific embodiments of the present invention does not limit the present invention. Those skilled in the art can 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 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 drug for treating non-Hodgkin's lymphoma, characterized in that: The anti-CD20 antibody drug conjugate has the following structure: Wherein, mAb represents a recombinant anti-CD20 monoclonal antibody, the drug-antibody ratio DAR is 4.2±1, preferably 4.2±0.5; and the at least one therapeutic agent is lenalidomide.

2. The use according to claim 1, characterized in that The non-Hodgkin's lymphoma is relapsed or refractory non-Hodgkin's lymphoma.

3. The use according to claim 2, characterized in that 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 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 any one of claims 1 to 5, characterized in that: The non-Hodgkin 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, precursor (peripheral) large granular lymphocytic leukemia or anaplastic large cell lymphoma.

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

8. The use according to claim 7, characterized in that The diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

9. The use according to claim 8, characterized in that The CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with dual expression of MYC and BCL2.

10. The use according to any one of claims 1 to 9, characterized in that: 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, 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.

12. The use according to claim 11, characterized in that The at least one standard treatment regimen is R-CHOP.

13. A drug combination for treating non-Hodgkin's lymphoma, characterized in that: The drug combination is a combination of an anti-CD20 antibody drug conjugate and lenalidomide, and the anti-CD20 antibody drug conjugate has the following structure: Wherein, mAb represents a recombinant anti-CD20 monoclonal antibody, and the drug-antibody ratio DAR is 4.2±1, preferably 4.2±0.

5.

14. The pharmaceutical combination according to claim 13, characterized in that The non-Hodgkin's lymphoma is relapsed or refractory non-Hodgkin's lymphoma.

15. The pharmaceutical combination according to claim 14, characterized in that The non-Hodgkin's lymphoma is relapsed or refractory non-Hodgkin's lymphoma after receiving at least one standard treatment regimen.

16. The pharmaceutical combination according to any one of claims 13 to 15, 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.

17. The pharmaceutical combination according to claim 16, characterized in that The recombinant anti-CD20 monoclonal antibody is selected from rituximab or its biosimilars.

18. A combination therapy for treating non-Hodgkin's lymphoma, characterized in that: An effective dose of the pharmaceutical combination according to any one of claims 13 to 17 is administered to a subject.

19. The combination therapy according to claim 18, characterized in that The non-Hodgkin 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, precursor (peripheral) large granular lymphocytic leukemia or anaplastic large cell lymphoma.

20. The combination therapy according to claim 19, characterized in that The non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.

21. The combination therapy according to claim 20, characterized in that The diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

22. The combination therapy according to claim 21, characterized in that The CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with dual expression of MYC and BCL2.

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

24. The combination therapy according to claim 23, 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.

25. The combination therapy according to claim 24, characterized in that The at least one standard treatment regimen is R-CHOP.

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