Pharmaceutical combination comprising Anti-CD20 antibody-drug conjugate and use thereof for treating non-hodgkin lymphoma

Abstract

An anti-CD20 antibody-drug conjugate, use thereof in combination with a therapeutic agent for treating relapsed or refractory non-Hodgkin lymphoma, and a composition thereof. The present invention relates to the field of biopharmaceuticals. The combined use of the anti-CD20 antibody-drug conjugate and at least one additional therapeutic agent has better efficacy than existing clinical second-line standard therapies.

Description

Pharmaceutical combinations comprising anti-cd20 antibody drug conjugates and their use in treating non-hodgkin lymphoma TECHNICAL FIELD

[0001] The present application relates to the field of pharmaceutical and pharmaceutical combination technology, in particular to the use of pharmaceutical combinations comprising anti-CD20 antibody drug conjugates in treating NHL. BACKGROUND

[0002] Non-Hodgkin lymphoma (NHL) is the seventh most common cancer worldwide, accounting for about 4-5% of new cancer cases and 3-4% of cancer-related deaths, affecting about 1.5 million people worldwide.

[0003] NHL is a collective term for a group of malignant proliferative diseases of the lymphatic system, including numerous types of lymphocytic tumors, which mainly originate from B lymphocytes (>85%), and a small number from T cells and natural killer cells. It is usually classified as slow progression or indolent NHL (iNHL) and rapid progression or aggressive NHL (aNHL) according to prognosis; among them, follicular lymphoma (FL) is the most common iNHL, accounting for about 1 / 5 of total NHL patients, while diffuse large B-cell lymphoma (DLBCL) is the most common aNHL, accounting for about 1 / 3 of total NHL patients.

[0004] 30-35% of DLBCL express MYC protein in DLBCL, 20-35% express BCL2 at the same time, called "double expression lymphoma", and part of DLBCL express MYC, BCL2 and BCL6 at the same time, called "triple expression lymphoma", which indicates poor prognosis.

[0005] The current clinical guidelines recommend the first-line standard treatment regimen for DLBCL including R-CHOP regimen, R-CHOEP regimen, R-miniCHOP regimen, DA-EPOCH-R regimen and Pola-R-CHP regimen, etc. The R-CHOP regimen uses rituximab (R), cyclophosphamide (C), vincristine (H), doxorubicin (O) and prednisone (P) in combination; the R-CHOEP regimen uses rituximab (R), cyclophosphamide (C), vincristine (H), doxorubicin (O), etoposide (E) and prednisone (P) in combination; the R-miniCHOP regimen uses rituximab (R), cyclophosphamide (C), vincristine (H), doxorubicin (O) and prednisone (P) in combination, wherein 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 dose adjustment (DA) is performed after each chemotherapy cycle according to the patient's response to chemotherapy; Pola-R-CHP uses polatuzumab vedotin (an ADC targeting CD79b, trade name ), rituximab (R), cyclophosphamide (C), vincristine (H) and prednisone (P) in combination. Although the patients have different degrees of remission after the above first-line therapy, 40%-50% of them are prone to relapse or refractory, especially for "double expression lymphoma" or "triple expression lymphoma", which shows poor prognosis.

[0006] The current second-line standard therapy for relapsed or refractory DLBCL after the first-line standard treatment regimen includes R-GemOx (rituximab (R), gemcitabine (Gem) and oxaliplatin (Ox) in combination), R 2 (rituximab (R) and lenalidomide (R) in combination), BR (bendamustine (B) and rituximab (R) in combination), R-ICE (rituximab, ifosfamide, carboplatin and etoposide in combination), R-DHAP (rituximab, cisplatin, cytarabine and dexamethasone in combination), R-GDP (rituximab, gemcitabine, cisplatin and dexamethasone in combination), R-MINE (rituximab, mesna, ifosfamide, mitoxantrone and etoposide in combination), R-ESHAP (rituximab, etoposide, methylprednisolone, cisplatin and cytarabine in combination), DA-EPOCH-R (rituximab, etoposide, prednisone, vincristine, cyclophosphamide and doxorubicin in combination). Although the patients have a certain degree of remission after the above second-line standard treatment regimen, there is still room for further improvement in clinical benefit, and therefore, there is still a need to find more effective treatment or combination therapy drugs and treatment regimens for relapsed or refractory non-Hodgkin's lymphoma after receiving at least one standard treatment regimen. SUMMARY

[0007] The present application aims to provide a pharmaceutical product / composition and its use in the preparation of a medicament for treating non-Hodgkin lymphoma, which has a better efficacy compared to the existing clinical second-line standard therapy.

[0008] One aspect of the present application discloses the use of a combination of an anti-CD20 antibody drug conjugate and at least another therapeutic agent in the preparation of a medicament for treating relapsed or refractory non-Hodgkin lymphoma.

[0009] Another aspect of the present application discloses the use of an anti-CD20 antibody drug conjugate and at least another therapeutic agent in the treatment of relapsed or refractory non-Hodgkin lymphoma.

[0010] Still another aspect of the present application discloses a method for treating relapsed or refractory non-Hodgkin lymphoma, which comprises administering to a subject in need thereof a therapeutically effective amount of an anti-CD20 antibody drug conjugate and at least another therapeutic agent.

[0011] The anti-CD20 antibody drug conjugate has the following structural formula:

[0012] mAb-(L-D)n;

[0013] 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 conjugated small molecule toxins to the monoclonal antibody; and “-” is a bond.

[0014] The preferred recombinant anti-CD20 monoclonal antibody includes but is not limited to Rituximab or its biosimilars, Ofatumumab or its biosimilars, Obinutuzumab or its biosimilars, Ocrelizumab or its biosimilars.

[0015] More preferably, the recombinant anti-CD20 monoclonal antibody is preferably Rituximab or its biosimilars.

[0016] The preferred D is one or more monomethyl auristatin, which includes but is not limited to monomethyl auristatin-E (MMAE), monomethyl auristatin-D (MMAD), or monomethyl auristatin-F (MMAF); further, the monomethyl auristatin is monomethyl auristatin-E (MMAE).

[0017] L is a cleavable linker or a non-cleavable linker, the non-cleavable linker includes but is not limited to a thioether bond linker, the thioether bond linker comprises a maleimide group, preferably the maleimide group further comprises hexanoic acid, forming a maleimide group hexanoyl (mc); preferably the maleimide group comprises a maleimide group methyl, for example succinimidyl-4-(N-maleimide group methyl) cyclohexane-1-carboxylate (sMCC) or sulfosuccinimidyl-4-(N-maleimide group methyl) cyclohexane-1-carboxylate (sulfo-sMCC).

[0018] The cleavable linker includes but is not limited to a hydrazone bond linker, a disulfide bond linker, a peptide bond linker; the peptide bond linker comprises valine-citrulline (Val-Cit or VC), phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala) or valine-lysine (Val-Lys). Preferably, the peptide bond linker is valine-citrulline (Val-Cit).

[0019] Further, the linker moiety further comprises p-aminobenzyl alcohol (PAB), p-aminobenzyloxycarbonyl (PABC) or derivatives or analogues thereof.

[0020] Further, L is preferably MC-VC-PAB.

[0021] n is an integer or a non-integer from 1 to 8; preferably n is 4.2±1, further preferably 4.2±0.5, and more preferably 4.2±0.3.

[0022] The anti-CD20 antibody drug conjugate further has the following structure:

[0023] wherein mAb represents a recombinant anti-CD20 monoclonal antibody, the average number of conjugation n of the small molecule toxin conjugated to the antibody is 1-8; preferably n is 4.2±1, further preferably n is 4.2±0.5, and more preferably n is 4.2±0.3. Those skilled in the art should understand that the average number of conjugation n of the small molecule toxin conjugated to the antibody can also be expressed as the drug antibody ratio DAR.

[0024] The anti-CD20 antibody drug conjugate of the present application can 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.

[0025] The carrier or excipient is selected from the group consisting of a pH buffer, an osmotic pressure regulator, a lyophilized powder excipient, a protein protective agent, a solubilizer, water for injection.

[0026] The pH buffer comprises hydrochloric acid histidine buffer, acetic acid buffer, phosphate buffer, citric acid buffer or Tris buffer, preferably the pH buffer is hydrochloric acid histidine buffer.

[0027] The protein protective agent comprises trehalose, sucrose, lactose or glucose, preferably the protein protective agent is trehalose.

[0028] The osmotic pressure regulator comprises mannitol or sodium chloride, preferably the osmotic pressure regulator is mannitol.

[0029] The solubilizer comprises polysorbate 80, polysorbate 20 or poloxamer 188, preferably the solubilizer is polysorbate 80.

[0030] Further, the liquid or lyophilized formulation comprises the anti-CD20 antibody drug conjugate, hydrochloric acid histidine, trehalose, mannitol and polysorbate 80.

[0031] In some embodiments, the recombinant anti-CD20 monoclonal antibody is selected from Rituximab or a biosimilar thereof, Ofatumumab or a biosimilar thereof, Ocrelizumab or a biosimilar thereof, Obinutuzumab or a biosimilar thereof.

[0032] Further, the recombinant anti-CD20 monoclonal antibody is selected from Rituximab or a biosimilar thereof.

[0033] In some embodiments, the at least another therapeutic agent is selected from one or more of a hormone drug, a platinum drug and an antimetabolite drug; the hormone drug is selected from one or more of prednisone, dexamethasone and methylprednisolone; the platinum drug is selected from one or more of carboplatin, cisplatin and oxaliplatin; the antimetabolite drug is selected from one or more of pemetrexed, methotrexate, fluorouracil, capecitabine, cytarabine, gemcitabine, 6-mercaptopurine and thioguanine.

[0034] Further, the at least another therapeutic agent can comprise at least one of gemcitabine, dexamethasone and cisplatin.

[0035] In some embodiments, 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, mantle cell lymphoma, precursor (peripheral) large granular lymphocytic leukemia or anaplastic large cell lymphoma.

[0036] Further, the non-Hodgkin lymphoma is diffuse large B-cell lymphoma.

[0037] Further, the diffuse large B-cell lymphoma is CD20 positive diffuse large B-cell lymphoma.

[0038] Further, the CD20 positive diffuse large B-cell lymphoma is MYC and BCL2 double-expressed diffuse large B-cell lymphoma.

[0039] In some embodiments, the relapsed or refractory non-Hodgkin lymphoma has received treatment of at least one standard treatment regimen.

[0040] In some embodiments, the subject has received treatment of at least one standard treatment regimen.

[0041] In some embodiments, the at least one standard treatment regimen comprises at least anti-CD20 antibody monotherapy or anti-CD20 antibody combination therapy.

[0042] Further, the at least one standard treatment regimen comprises at least one selected from R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R and Pola-R-CHP.

[0043] Further, the at least one standard treatment regimen is R-CHOP.

[0044] Another aspect of the present application discloses a composition for treating relapsed or refractory non-Hodgkin lymphoma, the composition comprising an anti-CD20 antibody drug conjugate and at least another therapeutic agent, the anti-CD20 antibody drug conjugate having the following structure:

[0045] wherein, mAb represents a recombinant anti-CD20 monoclonal antibody, the drug-antibody ratio (DAR) is 1-8, preferably the DAR is 4.2±1, further preferably the DAR is 4.2±0.5, and more further preferably the DAR is 4.2±0.3,

[0046] wherein, the at least another therapeutic agent is selected from one or more of a hormone drug, a platinum drug and an antimetabolite drug;

[0047] the hormone drug is selected from one or more of prednisone and dexamethasone;

[0048] the platinum drug is selected from one or more of carboplatin, cisplatin and oxaliplatin;

[0049] the antimetabolite drug is selected from one or more of pemetrexed, methotrexate, fluorouracil, capecitabine, cytarabine, gemcitabine, 6-mercaptopurine and thioguanine.

[0050] In some embodiments, the recombinant anti-CD20 mAb is selected from the group consisting of Rituximab (Rituxan®) or a biosimilar thereof, Ofatumumab (Arzerra®) or a biosimilar thereof, Ocrelizumab (Ocrevus®) or a biosimilar thereof, and Obinutuzumab (Gazyva®) or a biosimilar thereof.

[0051] Further, the recombinant anti-CD20 mAb is selected from the group consisting of Rituximab (Rituxan®) or a biosimilar thereof.

[0052] In some embodiments, the at least one additional therapeutic agent is gemcitabine, dexamethasone, and cisplatin.

[0053] In some embodiments, the composition is a combination of the anti-CD20 antibody drug conjugate, gemcitabine, dexamethasone, and cisplatin.

[0054] The combination of the anti-CD20 antibody drug conjugate and the at least one additional therapeutic agent described above is only an exemplary combination and does not constitute a limitation on the use of the anti-CD20 antibody drug conjugate of the present application in combination with other drugs.

[0055] The composition of the present application consists of a first pharmaceutical product comprising the anti-CD20 antibody drug conjugate and a carrier or excipient thereof and a second pharmaceutical product comprising the at least one additional therapeutic agent and a carrier or excipient thereof.

[0056] The first pharmaceutical product and the second pharmaceutical product of the present application can be packaged independently or co-packaged.

[0057] The present application also discloses a kit comprising the anti-CD20 antibody drug conjugate and the at least one additional therapeutic agent.

[0058] Further, the anti-CD20 antibody drug conjugate and the at least one additional therapeutic agent are packaged independently and disposed in the kit.

[0059] Further, the kit comprises the anti-CD20 antibody drug conjugate, gemcitabine, dexamethasone, and cisplatin.

[0060] Yet another aspect of the present application discloses a combination therapy for treating relapsed or refractory non-Hodgkin lymphoma after receiving at least one standard treatment regimen.

[0061] The combination therapy of the present application comprises administering an effective amount of the composition of the present application for treating relapsed or refractory non-Hodgkin lymphoma.

[0062] In some embodiments, the combination therapy described herein comprises administering an effective amount of the composition described herein for treating relapsed or refractory non-Hodgkin's lymphoma after receiving at least one standard treatment regimen.

[0063] In some embodiments, the combination therapy described herein comprises administering an effective amount of the anti-CD20 antibody drug conjugate, gemcitabine, dexamethasone, and cisplatin in combination.

[0064] In some embodiments, the non-Hodgkin's lymphoma is selected from small lymphocytic lymphoma, B-cell chronic lymphocytic leukemia, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, mantle cell lymphoma, precursor (peripheral) large granular lymphocyte leukemia, or anaplastic large cell lymphoma.

[0065] Further, the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.

[0066] Further, the diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

[0067] Further, the CD20-positive diffuse large B-cell lymphoma is MYC and BCL2 double-expressed diffuse large B-cell lymphoma.

[0068] In some embodiments, the at least one standard treatment regimen comprises at least anti-CD20 antibody monotherapy or anti-CD20 antibody combination therapy.

[0069] Further, the at least one standard treatment regimen comprises at least one selected from R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R, and Pola-R-CHP.

[0070] Further, the at least one standard treatment regimen is R-CHOP. It is understood by one skilled in the art that the R-CHOP regimen employs rituximab (R), cyclophosphamide (C), vincristine (H), doxorubicin (O), and prednisone (P) in combination.

[0071] In some embodiments, the anti-CD20 antibody drug conjugate is administered prior to, concurrently with, and / or after the at least another therapeutic agent.

[0072] In some embodiments, the anti-CD20 antibody drug conjugate and the at least another therapeutic agent can be administered in a single administration or multiple administrations. For example, the anti-CD20 antibody drug conjugate can be administered in a single administration or multiple administrations. For multiple administrations, the frequency of administration can be, for example, four times a week, three times a week, two times a week, one time a week, one time every two weeks, one time every three weeks, one time every four weeks, one time every five weeks, one time every six weeks, one time every seven weeks, or one time every eight weeks, etc. Alternatively, the administration can be in a cycle of one day, one week, two weeks, three weeks, four weeks, one month, two months, three months, six months, etc., with one, two, three, four, five, six, etc. administrations per cycle, and the administration can continue until the patient's disease progresses, dies, or is cured.

[0073] The at least another therapeutic agent, for example, gemcitabine, dexamethasone, and cisplatin, can be administered in a single administration or multiple administrations. For multiple administrations, the frequency of administration can be, for example, three times a day, two times a day, one time a day, one time every two days, one time a week, one time every two weeks, one time every three weeks, one time every four weeks, etc. Alternatively, the administration can be in a cycle of one day, one week, two weeks, three weeks, four weeks, one month, two months, three months, six months, etc., with one, two, three, four, five, six, etc. administrations per cycle, and the administration can continue until the patient's disease progresses, dies, or is cured.

[0074] The combination drug of the present application can be administered by any suitable means, including gastrointestinal, parenteral, intrapulmonary, and intranasal, etc. Parenteral infusions include intramuscular, intravenous, intra-arterial, intraperitoneal, or subcutaneous administration, etc.

[0075] Another aspect of the present application discloses a non-therapeutic method for inhibiting the proliferation of non-Hodgkin's lymphoma cells.

[0076] The method comprises adding an effective amount of the composition of the present application to a non-Hodgkin's lymphoma cell system that has received at least one inhibiting treatment.

[0077] In some embodiments, the method comprises administering an effective amount of the anti-CD20 antibody drug conjugate, gemcitabine, dexamethasone, and cisplatin to the non-Hodgkin's lymphoma cell system that has received at least one inhibiting treatment.

[0078] In some embodiments, the non-Hodgkin's lymphoma is selected from small lymphocytic lymphoma, B-cell chronic lymphocytic leukemia, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, mantle cell lymphoma, precursor (peripheral) large granular lymphocyte leukemia, or anaplastic large cell lymphoma.

[0079] Further, the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma. Further, the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.

[0080] Further, the diffuse large B-cell lymphoma is CD20 positive diffuse large B-cell lymphoma.

[0081] Further, the CD20 positive diffuse large B-cell lymphoma is MYC and BCL2 double expression diffuse large B-cell lymphoma.

[0082] In some embodiments, the at least one inhibiting treatment comprises at least one of anti-CD20 antibody monotherapy or anti-CD20 antibody combined with at least one other therapeutic agent.

[0083] Further, the at least one inhibiting treatment comprises treatment with at least one selected from R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R and Pola-R-CHP.

[0084] Further, the at least one inhibiting treatment is R-CHOP.

[0085] In another aspect of the present application, a container comprising the above composition is disclosed.

[0086] In another aspect of the present application, a kit comprising the above composition is disclosed.

[0087] For relapsed or refractory non-Hodgkin's lymphoma after receiving at least one standard treatment regimen, especially CD20 positive diffuse large B-cell lymphoma with BCL2 and MYC double expression, the anti-CD20 antibody drug conjugate combination regimen of the present application shows significant progress in tumor inhibition effect and better treatment potential compared with existing second-line standard therapy, and has good safety.

[0088] The anti-CD20 antibody drug conjugate of the present application has a synergistic effect with GDP in inhibiting tumor effect and improving drug efficacy.

[0089] The anti-CD20 antibody drug conjugate of the present application has a synergistic effect with GDP in inhibiting tumor effect and improving drug efficacy for non-Hodgkin's lymphoma patients or cells that have received R-CHOP inhibiting treatment. BRIEF DESCRIPTION OF DRAWINGS

[0090] Hereinafter, embodiments of the present application will be described in detail with reference to the accompanying drawings, in which:

[0091] Figure 1: Tumor volume change trend of mice in each administration group of WSU-DLCL2 / R-CHOP (relapse model);

[0092] Figure 2: Body weight change trend of mice in each administration group of WSU-DLCL2 / R-CHOP (relapse model). DETAILED DESCRIPTION

[0093] The present application will be described with respect to the following specific examples. Those skilled in the art will appreciate that these examples are intended for illustration only and are not intended to limit the scope of the present application.

[0094] The experimental methods in the following examples are all routine methods unless otherwise specified. The raw materials, reagents and materials used in the following examples are all commercially available unless otherwise specified.

[0095] Definitions:

[0096] "Rituximab" refers to: a CD20-targeted monoclonal antibody drug developed by Roche and approved for marketing by drug regulatory agencies with the trade name "Mabthera". Rituximab is its generic name, only the original drug can be called rituximab.

[0097] "Rituximab biosimilar" refers to: in addition to the original rituximab, other enterprises develop monoclonal antibody drugs with the same amino acid sequence as rituximab and similar physicochemical properties, efficacy, pharmacokinetics and safety as the original rituximab.

[0098] "High expression" refers to: the recognition of high expression of antigen or gene by technicians according to the detection results of tumor tissue or cell antigen or gene expression level and the evaluation standard higher than a certain expression level or relatively higher expression level, for example, strong positive (+++) and / or positive (++) determined according to the proportion of immunohistochemically colored cells and staining intensity.

[0099] "Low expression" refers to: the recognition of low expression of antigen or gene by technicians according to the detection results of tumor tissue or cell antigen or gene expression level and the evaluation standard lower than a certain expression level or relatively lower expression level, for example, weak positive (+) determined according to the proportion of immunohistochemically colored cells and staining intensity.

[0100] "Effective dose" or "effective amount" refers to an amount effective to achieve a desired therapeutic result at a desired dose and time period. The effective dose can vary depending on factors such as the individual's disease state, age, sex, and weight, and the ability of the therapeutic agent or combination of therapeutic agents to elicit the desired response in the individual.

[0101] The "standard treatment" referred to in the present application is not limited to the standard treatment regimen determined in the general sense of clinical practice, but also includes clinical recommended treatment regimen, clinical trial treatment regimen and other treatment regimens.

[0102] Although the present application has been described generally, embodiments of the present application will be further disclosed in the following examples, which should not be construed as limiting the scope of the claims.

[0103] Example 1: Preparation of anti-CD20 ADC (TRS005)

[0104] The preparation method of the exemplary anti-CD20 ADC (TRS005) is specifically described in Chinese Patent CN108452318B, which is incorporated herein in its entirety.

[0105] TRS005 has the following structure:

[0106] wherein mAb is rituximab or a rituximab biosimilar, and the drug-antibody ratio (DAR) is 4.2±0.5.

[0107] Example 2: Anti-tumor efficacy evaluation of TRS005 alone or in combination in a human lymphoma WSU-DLCL2 / R-CHOP relapse model

[0108] (I) Model establishment

[0109] The establishment of the relapse / refractory model uses conventional methods in the art. Briefly, WSU-DLCL2 cells are inoculated into SCID mice at a cell inoculation amount of 4*10 6 / each, treated with R-CHOP, defined as P1; tumor tissues with good growth state of WSU-DLCL2 / R-CHOP (P1) are selected, the fascia and internal necrotic tissues of the mice are removed, and then the tumor tissues are inoculated into SCID mice under sterile conditions, treated with R-CHOP, defined as P2; tumor tissues with good growth state of WSU-DLCL2 / R-CHOP (P2) are selected, the fascia and internal necrotic tissues of the mice are removed, and then the tumor tissues are inoculated into SCID mice under sterile conditions, treated with R-CHOP, defined as P3; the R-CHOP relapse model is constructed by P1-P3 passages through R-CHOP treatment. The R-CHOP administration scheme is shown in Table 1.

[0110] Table 1. R-CHOP administration scheme

[0111] wherein IV represents intravenous injection; IP represents intraperitoneal injection; PO represents oral administration;

[0112] QW*2wks represents once a week, for 2 weeks;

[0113] QD*5 / wk*2wks represents once a day, for 2 weeks, with administration on the first 5 days of each week;

[0114] BIW*2wks represents twice a week, with an interval of 3 days between each administration, for 2 weeks.

[0115] The WSU-DLCL2 / R-CHOP (P3) well-grown tumor tissues were selected, and after the fascia and internal necrotic tissues of the mice were removed, the tissues were cut into 2x2x2 mm 3 Small pieces were reserved in RPMI 1640 basic medium for later use. After the SCID mice were anesthetized with isoflurane, the tissue pieces were inoculated into the right subcutaneous tissue of the SCID mice under sterile conditions using an 11G cannula needle for the following experiments.

[0116] (ii) Grouping and administration

[0117] When the average tumor volume reached 120-150 mm 3 , the SCID mice were randomly grouped according to tumor volume, and the difference in tumor volume between groups was less than 10% of the average. In this experiment, 48 SCID mice were divided into 8 groups, with 6 mice in each group. The day of grouping was recorded as day 0, and the animals were immediately administered after grouping according to their body weight. The grouping and administration scheme is shown in Table 2.

[0118] Table 2. Grouping and administration scheme

[0119] Among them, IV represents intravenous injection; IP represents intraperitoneal injection; PO represents oral administration;

[0120] QWx3wks represents once a week for 3 weeks;

[0121] QDx5 / wkx3wks represents once a day, 5 days a week for 3 weeks;

[0122] QDx4 / wkx3wks represents once a day, 4 days a week for 3 weeks;

[0123] BIWx3wks represents twice a week, with an interval of 3 days, for 3 weeks.

[0124] The body weight and tumor volume of the animals were measured twice a week during the experiment. The animals were observed and recorded for clinical symptoms once a day during the experiment. Clinical observation included the overall health of the animals, abnormal body weight, abnormal behavior, and other drug-related adverse reactions.

[0125] (iii) Evaluation index and statistical analysis

[0126] The tumor growth inhibition rate (%TGI TV ) was calculated using the formula: (1-TV T / TV C ) x 100%, where TV C is the average tumor volume of the negative control group, and TV T is the average tumor volume of the treatment group.

[0127] The formula for calculating the relative tumor volume (RTV) is: Vt / V0, where V0 is the tumor volume at the time of grouping and Vt is the tumor volume at each measurement.

[0128] The formula for calculating the relative tumor proliferation rate (%T / C RTV ) is: T RTV / C RTV x 100%, where T RTV is the RTV of the treatment group and C RTV is the RTV of the negative control group.

[0129] The formula for calculating the rate of change in animal body weight (%BWC) is: (BWt-BW0) / BW0 x 100%, where BWt is the animal body weight at each measurement and BW0 is the animal body weight at the time of grouping.

[0130] According to the Chinese NMPA Technical Guidelines for Nonclinical Studies of Cytotoxic Antitumor Drugs (November 2006), %T / C RTV ≤ 40% and P < 0.05 by statistical analysis is considered effective. If the number of drug-related animal deaths exceeds 20%, the drug dose is considered to have severe toxicity.

[0131] The experimental data are expressed as mean ± SEM, and the two-tailed t-test was used for comparison between groups. P < 0.05 is considered a significant difference, and P < 0.01 is considered a highly significant difference (Microsoft Excel 2007, Redmond, WA, USA).

[0132] (IV) Test Results

[0133] 1. Inhibition of tumor volume by the test substance

[0134] The experimental period was 35 days. On day 25, the tumor volume of the vehicle control group was greater than 2000 mm 3 , and the animals reached the end of the experiment and were euthanized. The data on day 25 were used for statistical analysis. The results of the study on the antitumor activity of TRS005 combined with GDP in the relapsed model of human lymphoma WSU-DLCL2 / R-CHOP are shown in Tables 3-5 and Figure 1.

[0135] Referring to Table 3 and Figure 1, in the single-dose administration group:

[0136] Group 5, TRS005 low, had an average tumor volume of 963.43 ± 88.63 mm 3 , and the tumor growth inhibition rate %TGI TV was 40.51% (P < 0.01); the relative tumor proliferation rate %T / C RTV was 60.59% (P < 0.001).

[0137] Group 6 TRS005 high, average tumor volume was 564.28 ± 92.11 mm 3 , tumor growth inhibition rate %TGI TV was 65.16% (P < 0.001); relative tumor proliferation rate %T / C RTV was 36.02% (P < 0.001).

[0138] In the combination administration group:

[0139] Group 2 R-CHOP administration group, average tumor volume was 1024.81 ± 85.61 mm 3 , tumor growth inhibition rate %TGI TV was 36.72% (P < 0.05); relative tumor proliferation rate %T / C RTV was 64.15% (P < 0.001).

[0140] Group 3 R low + GDP administration group, average tumor volume was 507.68 ± 96.44 mm 3 , tumor growth inhibition rate %TGI TV was 68.65% (P < 0.001); relative tumor proliferation rate %T / C RTV was 31.53% (P < 0.001).

[0141] Group 4 R high + GDP administration group, average tumor volume was 252.57 ± 82.91 mm 3 , tumor growth inhibition rate %TGI TV was 84.40% (P < 0.001); relative tumor proliferation rate %T / C RTV was 14.66% (P < 0.001).

[0142] Group 7 TRS005 low + GDP administration group, average tumor volume was 42.93 ± 23.77 mm 3 , tumor growth inhibition rate %TGI TV was 97.35% (P < 0.001); relative tumor proliferation rate %T / C RTV was 2.29% (P < 0.001).

[0143] Group 8 TRS005 high + GDP administration group, average tumor volume was 7.69 ± 7.69 mm 3 , tumor growth inhibition rate %TGI TV was 99.53% (P < 0.001); relative tumor proliferation rate %T / C RTV was 0.42% (P < 0.001).

[0144] Table 3. Anti-tumor activity of TRS005 in combination with GDP *: P<0.05 compared with the solvent group. **: P<0.01 compared with the solvent group. ***: P<0.001 compared with the solvent group.

[0145] Referring to Table 4, the significant analysis of the anti-tumor activity shows that there is a very significant difference (P<0.001) between the low-dose TRS005 administration group and the low-dose TRS005+GDP administration group. There is a very significant difference (P<0.001) between the high-dose TRS005 administration group and the high-dose TRS005+GDP administration group.

[0146] There is a very significant difference (P<0.01) between the R low+GDP administration group and the low-dose TRS005+GDP administration group. There is a significant difference (P<0.05) between the R high+GDP administration group and the high-dose TRS005+GDP administration group.

[0147] Table 4. Significant analysis of anti-tumor activity of TRS005 monotherapy or combination with GDP

[0148] The above results show that the anti-CD20 antibody drug conjugate of the present application in combination with GDP significantly improves the tumor inhibition rate compared with the anti-CD20 antibody drug conjugate monotherapy and the second-line standard therapy R+GDP, and achieves excellent efficacy.

[0149] 2. Effects of the test substances on the body weight of tumor-bearing animals

[0150] After administration, the effects of the test substances on the body weight of the WSU-DLCL2 / R-CHOP relapse model tumor-bearing animals are shown in Table 5 and FIG. 2. The results show that on day 25, the average body weight of the animals in each group changed by -2.01%-6.78% (-0.49g-1.52g) compared with the average body weight at the beginning of the experiment on day 0. The body weight changes of the mice in each treatment group were within the normal range, no drug-related mouse deaths occurred, and no other obvious drug-related adverse reactions were observed, indicating that the TRS005 monotherapy and each TRS005 combination administration scheme of the present application have good safety.

[0151] Table 5. Effects of administration of each group on the body weight of tumor-bearing animals

[0152] In summary, for the WSU-DLCL2 / R-CHOP relapse model, the tumor inhibition rates of the TRS005 high+GDP and TRS005 low+GDP two dose groups are significantly improved compared with the corresponding two-line standard therapy R high+GDP and R low+GDP two dose groups; the tumor inhibition rates of the TRS005 high+GDP and TRS005 low+GDP two dose groups are significantly improved compared with the corresponding TRS005 high and TRS005 low two single drug dose groups; no animal death and other drug-related side effects are observed during the experiment, and the TRS005 combined with GDP scheme of the present application has good safety.

[0153] The anti-CD20 antibody drug conjugate combined scheme of the present application shows a significantly improved tumor inhibition effect and better treatment potential compared with the existing two-line standard therapy, and has good safety; the anti-CD20 antibody drug conjugate combined with GDP has a synergistic effect in inhibiting tumor effect and improving drug efficacy after the inhibiting treatment of R-CHOP.

[0154] The above description of the specific embodiments of the present application does not limit the present application, and those skilled in the art can make various changes or modifications to the present application without departing from the spirit of the present application, and all such changes or modifications shall belong to the scope of the claims attached to the present application.

Claims

1. Use of an anti-CD20 antibody drug conjugate in combination with at least another therapeutic agent for the treatment of relapsed or refractory non-Hodgkin's lymphoma, wherein, The anti-CD20 antibody drug conjugate has the following structure: mAb, wherein mAb represents a recombinant anti-CD20 monoclonal antibody, the drug-antibody ratio (DAR) is 1-8, preferably the DAR is 4.2±1, more preferably the DAR is 4.2±0.5, and even more preferably the DAR is 4.2±0.3; wherein the at least another therapeutic agent comprises one or more of a hormone drug, a platinum drug or an anti-metabolite drug; the hormone drug comprises one or more of prednisone, dexamethasone or methylprednisolone; the platinum drug comprises one or more of carboplatin, cisplatin or oxaliplatin; the anti-metabolite drug comprises one or more of pemetrexed, methotrexate, fluorouracil, capecitabine, cytarabine, gemcitabine, 6-mercaptopurine or thioguanine.

2. The use of claim 1, wherein, the recombinant anti-CD20 monoclonal antibody is selected from Rituximab or a biosimilar thereof, Ofatumumab or a biosimilar thereof, Ocrelizumab or a biosimilar thereof, and Obinutuzumab or a biosimilar thereof; preferably, the recombinant anti-CD20 monoclonal antibody is Rituximab or a biosimilar thereof.

3. The use of claim 1, wherein, the at least another therapeutic agent comprises at least one of gemcitabine, dexamethasone and cisplatin.

4. The use of claim 1, wherein, 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, mantle cell lymphoma, precursor (peripheral) large granular lymphocytic leukemia and anaplastic large cell lymphoma; preferably, the non-Hodgkin lymphoma is diffuse large B-cell lymphoma; more preferably, the diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

5. The use of claim 4, wherein, the CD20-positive diffuse large B-cell lymphoma is MYC and BCL2 double-expressed diffuse large B-cell lymphoma.

6. The use of claim 1, wherein, the relapsed or refractory non-Hodgkin lymphoma has been treated with at least one standard treatment regimen, preferably, the at least one standard treatment regimen comprises at least anti-CD20 antibody monotherapy or anti-CD20 antibody combination therapy.

7. The use of claim 6, wherein, the at least one standard treatment regimen comprises one selected from R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R and Pola-R-CHP; preferably, the at least one standard treatment regimen comprises R-CHOP.

8. The use of claim 1, wherein, the administration of the antibody drug conjugate comprises single administration or multiple administration; for multiple administration, the administration frequency is four times a week, three times a week, two times a week, one time a week, one time two weeks, one time three weeks, one time four weeks, one time five weeks, one time six weeks, one time seven weeks or one time eight weeks, etc.; or one time, two times, three times, four times, five times or six times per cycle, wherein one cycle is one day, one week, two weeks, three weeks, four weeks, one month, two months or three months; and / or The administration of the therapeutic agent includes single administration or multiple administration. For multiple administration, the administration frequency can be, for example, three times a day, twice a day, once a day, once every two days, once a week, once every two weeks, once every three weeks, once every four weeks, etc. Alternatively, the administration can be once, twice, three times, four times, five times, six times per cycle, wherein the cycle is one day, one week, two weeks, three weeks, four weeks, one month, two months, three months, six months, etc.

9. The use of claim 1, wherein, The anti-CD20 antibody drug conjugate is administered before, simultaneously with, and / or after the at least one other therapeutic agent.

10. A composition for treating relapsed or refractory non-Hodgkin's lymphoma, wherein, The composition comprises an anti-CD20 antibody drug conjugate having the structure: wherein mAb represents a recombinant anti-CD20 monoclonal antibody, the drug-antibody ratio (DAR) is 1-8, preferably the DAR is 4.2±1, further preferably the DAR is 4.2±0.5, and more further preferably the DAR is 4.2±0.3; wherein the at least one other therapeutic agent is selected from one or more of a hormone agent, a platinum agent, and an antimetabolite agent; the hormone agent is selected from one or more of prednisone and dexamethasone; the platinum agent is selected from one or more of carboplatin, cisplatin, and oxaliplatin; the antimetabolite agent is selected from one or more of pemetrexed, methotrexate, fluorouracil, capecitabine, cytarabine, gemcitabine, 6-mercaptopurine, and thioguanine.

11. The composition of claim 10, wherein, the recombinant anti-CD20 monoclonal antibody is selected from Rituximab or a biosimilar thereof, Ofatumumab or a biosimilar thereof, Ocrelizumab or a biosimilar thereof, and Obinutuzumab or a biosimilar thereof.

12. The composition of claim 10, wherein, the recombinant anti-CD20 monoclonal antibody is selected from Rituximab or a biosimilar thereof.

13. The composition of claim 10, wherein, the at least one other therapeutic agent comprises at least one of gemcitabine, dexamethasone, and cisplatin.

14. The composition of claim 10, wherein, the relapsed or refractory non-Hodgkin lymphoma comprises a non-Hodgkin lymphoma that has received at least one standard treatment regimen, preferably, the at least one standard treatment regimen comprises at least one selected from R-CHOP, R-CHOEP, R-miniCHOP, DA-EPOCH-R, and Pola-R-CHP, more preferably R-CHOP.

15. A method of non-therapeutically inhibiting proliferation of non-Hodgkin's lymphoma cells, wherein, adding an effective dose of the composition of claim 10 to a non-Hodgkin lymphoma cell system, wherein the non-Hodgkin lymphoma cell is a relapsed or refractory non-Hodgkin lymphoma cell that has received at least one inhibitory treatment.

16. A container, wherein, comprising the composition of any one of claims 10-14.

17. A kit, wherein, comprising the composition of any one of claims 10-14.

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