Use of a pharmaceutical composition comprising an anti-CD20 antibody-drug conjugate in the manufacture of a treatment for NHL

Abstract

The present invention discloses the use of a pharmaceutical composition comprising an anti-CD20 antibody-drug conjugate in the manufacture of a therapeutic agent for non-Hodgkin's lymphoma, the pharmaceutical composition comprising an anti-CD20 antibody-drug conjugate and at least one therapeutic agent, and furthermore, the combination therapy using the pharmaceutical composition has superior therapeutic effects, fewer dosage types, shorter infusion times, lower safety risks, and a lower economic burden compared to existing clinical first-line standard therapies.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This patent application claims priority from Chinese Patent Application No. CN202310337027.9, filed on March 31, 2023, the entire contents of which are incorporated herein by reference.

[0002] Technical Field The present invention relates to the field of pharmaceuticals and pharmaceutical combinations, and in particular to the use of pharmaceutical compositions comprising anti-CD20 antibody-drug conjugates in the manufacture of drugs for the treatment of NHL. [Background technology]

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

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

[0005] Double expressor lymphoma (DEL) is defined in the 2016 World Health Organization (WHO) classification revision as overexpression of MYC and BCL-2 proteins independent of chromosomal rearrangements. DEL accounts for 20-30% of newly diagnosed DLBCL. DEL is a highly aggressive lymphoma with poor progression-free survival (PFS).

[0006] Clinical guidelines recommend first-line treatment for most NHL subtypes, including DLBCL, including the quadruple combination of rituximab (R) with cyclophosphamide (C), doxorubicin (H), vincristine (O), and prednisone (P) (R-CHOP). While the majority of NHL patients treated with R-CHOP demonstrate favorable remission rates, there is still room for improvement. Furthermore, this regimen involves as many as five different drugs, has long administration times, carries certain transfusion risks, and is expensive.

[0007] A recently approved first-line treatment regimen for DLBCL is Pola-R-CHP (i.e., replacing chemotherapy drug O in the R-CHOP regimen with a CD79b-ADC). This regimen does not reduce the number of combination drugs; it simply employs the costly ADC drug polatuzumab (POLIVY) to replace the inexpensive chemotherapy drug vincristine (O). Current first-line DLBCL clinical trials include various R-CHOP+X regimens, exploring the possibility of enhancing therapeutic efficacy based on R-CHOP. However, these regimens all rely on additive approaches based on R-CHOP, further increasing the number of drugs used. These regimens, on the one hand, increase drug risk for patients and, on the other hand, increase treatment costs.

[0008] Therefore, there is a need to explore new, more effective treatments and combination therapies for NHL, especially DLBCL, including drugs and treatment protocols that reduce the number of combination drugs and further improve therapeutic efficacy compared to standard therapies. Summary of the Invention [Problem to be solved by the invention]

[0009] The present invention provides a pharmaceutical composition and its use in the manufacture of a therapeutic agent for non-Hodgkin's lymphoma. The combination therapy using this pharmaceutical composition exhibits superior therapeutic effects compared to existing standard first-line clinical therapies, and requires fewer drugs, shortens infusion times, reduces safety risks, and reduces the economic burden. [Means for solving the problem]

[0010] A first aspect of the present invention discloses the use of an anti-CD20 antibody-drug conjugate, alone or in combination with at least one therapeutic agent, in the manufacture of a medicament for NHL.

[0011] The anti-CD20 antibody-drug conjugate has the following structural formula: mAb-(LD)n;

[0012] In the formula, mAb is a recombinant anti-CD20 monoclonal antibody, D is a small molecule toxin, L is a linker connecting the monoclonal antibody and the small molecule toxin, n is the average number of small molecule toxins bound to the monoclonal antibody, and "-" represents a bond.

[0013] Preferred recombinant anti-CD20 monoclonal antibodies include, but are not limited to, rituximab or a biologically similar drug thereof, ofatumumab or a biologically similar drug thereof, obinutuzumab or a biologically similar drug thereof, and ocrelizumab or a biologically similar drug thereof.

[0014] More preferably, the recombinant anti-CD20 monoclonal antibody is rituximab or a biologically similar drug thereof.

[0015] Preferably, D is one or more monomethyl auristatins, including, but not limited to, monomethyl auristatin-E (MMAE), monomethyl auristatin-D (MMAD), or monomethyl auristatin-F (MMAF); further, the monomethyl auristatin is monomethyl olestatin-E (MMAE).

[0016] L is a cleavable linker or a non-cleavable linker, including, but not limited to, a thioether-linked linker, which includes a maleimide group. Preferred maleimide groups include a maleimidocaproyl group (mc) that further contains caproic acid. Preferred maleimide groups also include maleimidomethyl groups, such as succinimide-4-(N-maleimidomethyl)cyclohexane-1-carboxylic acid ester (sMCC) or sulfosuccinimide-4-(N-maleimidomethyl)cyclohexane-1-carboxylic acid ester (sulfo-sMCC).

[0017] Cleavable linkers include, but are not limited to, hydrazone bond linkers, disulfide bond linkers, and peptide bond linkers, including valine-citrulline (Val-Cit or VC), phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), or valine-lysine (Val-Lys). A preferred peptide bond linker is valine-citrulline (Val-Cit).

[0018] Additionally, the linker moiety includes para-aminobenzyl (PAB), para-amino-benzyloxycarbonyl (PABC) or derivatives or analogs thereof.

[0019] Furthermore, L is preferably MC-VC-PAB.

[0020] n is an integer or a non-integer of 1 to 8; preferably, n is 4.2±1, more preferably 4.2±0.5, and even more preferably 4.2±0.3.

[0021] The anti-CD20 antibody-drug conjugate further has the structure: [ka]

[0022] In the formula, mAb is rituximab or a biologically similar drug thereof, and the average number n of small molecule toxins bound to the antibody is 1 to 8, preferably n is 4.2±1, more preferably n is 4.2±0.5, and even more preferably n is 4.2±0.3.

[0023] The anti-CD20 antibody-drug conjugate of the present invention may be a pharmaceutical product containing the CD20 antibody-drug conjugate and a carrier or excipient, and the pharmaceutical product may be a liquid or lyophilized preparation.

[0024] The carrier or excipient is selected from the group consisting of a pH buffering agent, an osmolality adjusting agent, a lyophilized powder excipient, a protein protecting agent, a solubilizing agent, and water for injection.

[0025] Examples of the pH buffer include histidine hydrochloride buffer, acetate buffer, phosphate buffer, citrate buffer, and Tris buffer, and a preferred pH buffer is histidine hydrochloride buffer.

[0026] The protein protecting agent includes trehalose, sucrose, lactose, or glucose, and a preferred protein protecting agent is sucrose.

[0027] The osmolality adjuster includes mannitol or sodium chloride, and a preferred osmolality adjuster is mannitol.

[0028] The solubilizing agent may be polysorbate 80, polysorbate 20, or poloxamer 188, and a preferred solubilizing agent is polysorbate 80.

[0029] The liquid or lyophilized formulation further comprises the anti-CD20 antibody-drug conjugate, histidine hydrochloride, trehalose, mannitol, and polysorbate 80.

[0030] the at least one therapeutic agent is one or more selected from the group consisting of alkylating agents, antibiotics, botanical preparations, hormone-based drugs, platinum preparations, and antimetabolites;

[0031] the alkylating agent is at least one selected from the group consisting of cyclophosphamide, ifosfamide, melphalan, busulfan, chlormethine, chlorambucil, thiotepa, bendamustine, carmustine, nimustine, lomustine, and semustine;

[0032] the antibiotic is one or more selected from the group consisting of doxorubicin, daunorubicin, bleomycin, mitomycin, epirubicin, aclarubicin, and idarubicin;

[0033] The plant component preparation is one or more selected from the group consisting of vinca alkaloids, vinblastine, vincristine, vindesine, vinorelbine, docetaxel, camptothecin, irinotecan, exatecan, topotecan, paclitaxel, and taxanes;

[0034] the hormone-based drug is one or more selected from the group consisting of prednisone and dexamethasone;

[0035] The platinum agent is one or more selected from the group consisting of carboplatin, cisplatin, and oxaliplatin;

[0036] The antimetabolite is one or more selected from the group consisting of pemetrexed, methotrexate, fluorouracil, capecitabine, cytarabine, gemcitabine, 6-mercaptopurine, and thioguanine.

[0037] Preferably, the at least one therapeutic agent is an alkylating agent, an antibiotic, and a hormonal agent;

[0038] the alkylating agent is at least one selected from the group consisting of cyclophosphamide, ifosfamide, melphalan, busulfan, chlormethine, chlorambucil, thiotepa, bendamustine, carmustine, nimustine, lomustine, and semustine;

[0039] the antibiotic is one or more selected from the group consisting of doxorubicin, daunorubicin, bleomycin, mitomycin, epirubicin, aclarubicin, and idarubicin;

[0040] The hormone-based drug is one or more selected from the group consisting of prednisone and dexamethasone.

[0041] Further, the at least one therapeutic agent is cyclophosphamide, doxorubicin, and prednisone, ie, the pharmaceutical composition is the anti-CD20 antibody-drug conjugate, cyclophosphamide, doxorubicin, and prednisone.

[0042] Preferably, the non-Hodgkin's lymphoma is selected from the group consisting of small lymphocytic lymphoma, B-cell chronic lymphocytic lymphoma, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, mantle cell lymphoma, precursor B-cell lymphoblastic leukemia / lymphoma (B-ALL / LBL), T-cell large granular lymphocytic leukemia or anaplastic large cell lymphoma (ALCL).

[0043] Furthermore, the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma, and a preferred diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

[0044] Furthermore, the diffuse large B-cell lymphoma is a newly diagnosed diffuse large B-cell lymphoma, and preferably, the newly diagnosed diffuse large B-cell lymphoma is a newly diagnosed CD20-positive diffuse large B-cell lymphoma.

[0045] Furthermore, the CD20-positive diffuse large B-cell lymphoma is diffuse large B-cell lymphoma with high CD20 expression or low CD20 expression.

[0046] Still further, the newly diagnosed CD20-positive diffuse large B-cell lymphoma is a newly diagnosed CD20-high or CD20-low expression diffuse large B-cell lymphoma.

[0047] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with double expression of MYC and BCL2 and high or low expression of CD20.

[0048] Furthermore, the newly diagnosed CD20-positive diffuse large B-cell lymphoma is a newly diagnosed diffuse large B-cell lymphoma that double expresses MYC and BCL2 and has high or low CD20 expression.

[0049] A preferred invention discloses the use of an anti-CD20 antibody-drug conjugate in combination with cyclophosphamide, doxorubicin and prednisone in the manufacture of a medicament for the treatment of CD20-positive diffuse large B-cell lymphoma.

[0050] Additionally, the present invention discloses the use of an anti-CD20 antibody-drug conjugate in combination with cyclophosphamide, doxorubicin, and prednisone in the manufacture of a medicament for the treatment of newly diagnosed CD20-positive diffuse large B-cell lymphoma.

[0051] Furthermore, the present invention discloses the use of an anti-CD20 antibody-drug conjugate in combination with cyclophosphamide, doxorubicin, and prednisone in the manufacture of a medicament for the treatment of CD20 high or CD20 low expressing diffuse large B-cell lymphoma.

[0052] Still further, the present invention discloses the use of an anti-CD20 antibody-drug conjugate in combination with cyclophosphamide, doxorubicin, and prednisone in the manufacture of a medicament for the treatment of newly diagnosed CD20-high or CD20-low diffuse large B-cell lymphoma.

[0053] A second aspect of the present invention discloses a medicament for use in the treatment of non-Hodgkin's lymphoma.

[0054] The medicament comprises an anti-CD20 antibody-drug conjugate and at least one therapeutic agent, wherein the anti-CD20 antibody-drug conjugate has a structure according to the first aspect of the invention and the at least one therapeutic agent is of a type according to the first aspect of the invention.

[0055] Preferably, the non-Hodgkin's lymphoma is selected from the group consisting of small lymphocytic lymphoma, B-cell chronic lymphocytic lymphoma, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, mantle cell lymphoma, precursor B-cell lymphoblastic leukemia / lymphoma (B-ALL / LBL), T-cell large granular lymphocytic leukemia or anaplastic large cell lymphoma (ALCL).

[0056] Furthermore, the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma, and a preferred diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

[0057] Furthermore, the diffuse large B-cell lymphoma is a newly diagnosed diffuse large B-cell lymphoma, and preferably, the newly diagnosed diffuse large B-cell lymphoma is a newly diagnosed CD20-positive diffuse large B-cell lymphoma.

[0058] Furthermore, the CD20-positive diffuse large B-cell lymphoma is diffuse large B-cell lymphoma with high CD20 expression or low CD20 expression.

[0059] Still further, the newly diagnosed CD20-positive diffuse large B-cell lymphoma is a newly diagnosed CD20-high or CD20-low expression diffuse large B-cell lymphoma.

[0060] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with double expression of MYC and BCL2 and high or low expression of CD20.

[0061] Furthermore, the newly diagnosed CD20-positive diffuse large B-cell lymphoma is a newly diagnosed diffuse large B-cell lymphoma that double expresses MYC and BCL2 and has high or low CD20 expression.

[0062] Preferably, the pharmaceutical agent used to treat the non-Hodgkin's lymphoma comprises the anti-CD20 antibody-drug conjugate, cyclophosphamide, doxorubicin, and prednisone.

[0063] A third aspect of the present invention discloses a method for treating non-Hodgkin's lymphoma.

[0064] The treatment method of the present invention comprises administering to a subject an effective amount of the anti-CD20 antibody-drug conjugate or the medicament according to the second aspect of the invention for the treatment of non-Hodgkin's lymphoma, wherein the anti-CD20 antibody-drug conjugate in the medicament may precede, coincide with and / or follow administration of at least one therapeutic agent.

[0065] Preferably, the non-Hodgkin's lymphoma is selected from the group consisting of small lymphocytic lymphoma, B-cell chronic lymphocytic lymphoma, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, mantle cell lymphoma, precursor B-cell lymphoblastic leukemia / lymphoma (B-ALL / LBL), T-cell large granular lymphocytic leukemia or anaplastic large cell lymphoma (ALCL).

[0066] Furthermore, the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma, and a preferred diffuse large B-cell lymphoma is CD20-positive diffuse large B-cell lymphoma.

[0067] Furthermore, the diffuse large B-cell lymphoma is a newly diagnosed diffuse large B-cell lymphoma, and preferably, the newly diagnosed diffuse large B-cell lymphoma is a newly diagnosed CD20-positive diffuse large B-cell lymphoma.

[0068] Furthermore, the CD20-positive diffuse large B-cell lymphoma is diffuse large B-cell lymphoma with high CD20 expression or low CD20 expression.

[0069] Still further, the newly diagnosed CD20-positive diffuse large B-cell lymphoma is a newly diagnosed CD20-high or CD20-low expression diffuse large B-cell lymphoma.

[0070] Furthermore, the CD20-positive diffuse large B-cell lymphoma is a diffuse large B-cell lymphoma with double expression of MYC and BCL2 and high or low expression of CD20.

[0071] Furthermore, the newly diagnosed CD20-positive diffuse large B-cell lymphoma is a newly diagnosed diffuse large B-cell lymphoma that double expresses MYC and BCL2 and has high or low CD20 expression.

[0072] A preferred combination therapy of the present invention comprises the combined use of effective amounts of an anti-CD20 antibody-drug conjugate, cyclophosphamide, doxorubicin, and prednisone.

[0073] The combination of the present invention can be administered by any suitable means, including gastrointestinal, parenteral, pulmonary, intranasal, etc. Parenteral administration includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration.

[0074] A generally suitable method for preventing or treating a disease using the anti-CD20 antibody-drug conjugate of the present invention and at least one therapeutic agent is to administer the drug to a patient one or more times. The initial dose of the anti-CD20 antibody-drug conjugate administered to a patient may be about 1 μg / kg to 100 mg / kg, e.g., 0.1 mg / kg to 20 mg / kg, depending on the type and severity of the disease. For repeated administration over several days or more, treatment is generally continued until a desired suppression of disease symptoms occurs, depending on the circumstances. An exemplary dose range for the antibody-drug conjugate may be about 0.05 mg / kg to about 10 mg / kg.

[0075] The anti-CD20 antibody-drug conjugate of the present invention is administered to a patient multiple times, and the administration frequency may be once every 1 to 6 weeks, for example, once every 2, 3, or 4 weeks, and most preferably once every 3 weeks. The anti-CD20 antibody-drug conjugate and the at least one therapeutic agent are administered until the subject is cured, the disease progresses, or the subject dies.

[0076] A fourth aspect of the present invention discloses a non-therapeutic method of inhibiting non-Hodgkin's lymphoma cells.

[0077] The method comprises administering to a non-Hodgkin's lymphoma cell line an effective amount of a pharmaceutical agent according to the second aspect of the invention.

[0078] Preferably, the non-therapeutic method of inhibiting non-Hodgkin's lymphoma cells of the present invention comprises administering to a non-Hodgkin's lymphoma cell line effective amounts of an anti-CD20 antibody-drug conjugate, cyclophosphamide, doxorubicin, and prednisone.

[0079] It should be understood that one, some, or all of the various embodiments described herein may be combined to form other embodiments of the present invention. These and other embodiments of the present invention are further described in the detailed description that follows.

[0080] Hereinafter, an embodiment of the present invention will be described in detail with reference to the accompanying drawings. [Brief explanation of the drawings]

[0081] [Figure 1] : Tumors removed from WSU-DLCL2 cell xenograft tumor mouse models after mice from each treatment group were euthanized on day 35; [Figure 2] : Changes in tumor volume in each treatment group in a WSU-DLCL2 cell xenograft tumor mouse model; [Figure 3] : Changes in tumor growth inhibition rate in each treatment group in WSU-DLCL2 cell xenograft tumor mouse model; [Figure 4] : Trends in tumor volume change in mice for each treatment group in an OCI-LY19 cell xenograft tumor mouse model. DETAILED DESCRIPTION OF THE INVENTION

[0082] Specific Embodiments The present invention will now be described with reference to specific examples, which should be understood by those skilled in the art as being illustrative of the present invention and not limiting the scope of the present invention in any way.

[0083] The experimental methods in the following examples are conventional methods unless otherwise specified. Raw materials, reagents, etc. used in the following examples are commercially available products unless otherwise specified.

[0084] Definition:

[0085] "Rituximab" refers to a monoclonal antibody drug targeting CD20, developed by Roche and approved by the National Medical Products Regulatory Authority and marketed under the trade name "Mei Luohua." Rituximab is its generic name; only the original drug can be called rituximab.

[0086] "Biologically similar drugs to rituximab" refers to monoclonal antibody drugs, excluding the original drug rituximab, that have been developed by other companies to imitate rituximab, whose amino acid sequence is identical to that of rituximab and whose physicochemical properties, therapeutic effects, pharmacokinetics and safety are similar to those of the original drug rituximab.

[0087] "High CD20 expression" means that a technician determines, based on the results of detecting the CD20 expression level in tumor tissues or cells, that an expression level higher than or relatively high than a certain expression level based on conventional evaluation criteria is high CD20 expression. For example, high CD20 expression can be determined from CD20 strongly positive (+++) and / or positive (++) determined based on the proportion and staining intensity of immunohistochemically stained cells.

[0088] "Low CD20 expression" refers to a condition in which a technician determines the CD20 expression level of tumor tissue or cells based on the results of detection, and determines that the expression level is below a certain level or is relatively low based on conventional evaluation criteria, for example, that the CD20 expression level is weakly positive (+) for CD20 determined based on the proportion of immunohistochemically stained cells and staining intensity.

[0089] An "effective dose" refers to an amount effective, at a desired dosage and for a desired period of time, to achieve a desired therapeutic result. An effective dose can vary depending on factors such as the disease state, age, sex, and weight of the individual, and the ability of the therapeutic agent or combination of therapeutic agents to elicit a desired response in the individual.

[0090] Having generally described the present invention, the following examples further disclose embodiments of the invention and are not to be construed as limiting the scope of the claims.

[0091] Example 1: Preparation of anti-CD20 ADC (TRS005) For an exemplary method for producing anti-CD20 ADC (TRS005), see patent application CN108452318A, the entire text of which is incorporated herein by reference.

[0092] The structure of TRS005 is as follows:

[0093] [ka]

[0094] Here, the mAb is rituximab or a biosimilar of rituximab with a drug-antibody ratio (DAR) of 4.2±0.5.

[0095] Example 2: Screening of human lymphoma cell lines

[0096] The human diffuse large B-cell lymphoma cell line WSU-DLCL2, which has high expression of BCL-2 and MYC and high expression of CD20, and the human diffuse large B-cell lymphoma cell line OCI-LY19, which has high expression of BCL-2 and MYC and low expression of CD20, were screened by measuring the expression levels (mRNA levels) of BCL-2, MYC, and CD20 in each human diffuse large B-cell lymphoma cell line. The expression levels of CD20, BCL2, and MYC in both cell lines are shown in Table 1.

[0097] Table 1: Expression levels of CD20, BCL2, and MYC in both cell lines (log2TPM).

[0098] [Table 1]

[0099] Example 3: Study of the growth inhibitory effect of TRS005 alone and in combination with CHOP compared with R-CHOP regimen on human lymphoma WSU-DLCL2 cell xenograft tumors

[0100] Drug information, model building, grouping and administration The TRS005 formulation was provided by Zhejiang Tesco Pharmaceutical Co., Ltd., and all other positive drugs were commercially available, of which MabThera, cyclophosphamide, doxorubicin, vincristine, and prednisone were formulated to the appropriate concentrations using saline as the solvent and stored at 4°C away from light. The TRS005 formulation was reconstituted with sterile water for injection, and the corresponding volume of saline was added to adjust to the corresponding concentration. The TRS005 formulation was reconstituted with water for injection, packaged, and stored at -70°C.

[0101] Under sterile conditions, a concentration of 410 7 0.1 mL of the WSU-DLCL2 cell suspension was inoculated subcutaneously into the right dorsal region. The average tumor volume was approximately 150 mm 3 When tumor volume reached 1000 mg / kg, the animals were randomly divided into four groups according to tumor volume: Group 1 vehicle group, Group 2 R-CHOP group, Group 3 TRS005 single agent group, and Group 4 T-CHP group. Each group contained six mice, and tumor volume differences were less than 10% of the mean. The day of grouping was designated Day 0, and administration began according to the animals' weight. The specific grouping and administration regimen are shown in Table 2:

[0102] Table 2. Grouping and administration plan [Table 2]

[0103] where IV = intravenous injection; IP = intraperitoneal injection; PO = oral administration; BIW x 3w = twice a week for a total of 3 weeks; QW x 3w = once a week for a total of 3 weeks; QD x 5 / w x 3w = once a day, 5 times a week for a total of 3 weeks.

[0104] During the study, animal weights and tumor volumes were measured twice weekly. Clinical observations were recorded daily, including the animals' overall health, weight and behavioral abnormalities, and any other treatment-related adverse reactions.

[0105] Evaluation indicators The tumor growth inhibition rate (%TGI) was (1-TV T / TV C )×100%, TV C is the mean tumor volume in the negative control group, TV T was calculated based on the mean tumor volume of the treatment group.

[0106] Relative tumor volume (RTV) is V t / V0, where V0 is the tumor volume at the time of grouping, and V t is the tumor volume at each measurement.

[0107] Relative tumor growth rate (%T / C RTV ) is T RTV / C RTV The calculation was done by multiplying the T by 100%. RTV is the treatment group RTV, and C RTV is the negative control group RTV.

[0108] The percent weight change (%BWC) of an animal is (BW t -BW0) / BW0×100% t is the body weight of the animal at each measurement, and BW0 is the body weight of the animal at the time of grouping.

[0109] According to the China NMPA "Technical Guidelines for Non-clinical Research on Cytotoxic Anticancer Drugs" (November 2006), %T / C RTVA statistical analysis of ≦40% and P<0.05 was considered significant. If the drug-related animal mortality rate exceeded 20%, the drug dose was considered to be severely toxic.

[0110] Test results (1) Tumor-inhibiting effect In this experiment, the effects of single agent or combined administration of TRS005 on the tumor volume of human lymphoma WSU-DLCL2 are shown in Table 3 and Figures 1-3.

[0111] Table 3. Effect of administration treatment on tumor volume in each group

[0112] [Table 3]

[0113] Note: *** indicates P<0.001 compared to Group 1. ### indicates P<0.001 compared to Group 2. $ indicates P<0.05 compared to Group 3.

[0114] As can be seen from Table 3 and Figures 1-3, the mean tumor volume in Group 4 (TRS005-CHP) on Day 35 was 65.73 ± 26.35 mm 3 and Group 2 (R-CHOP) 782.94±94.87mm 3 Group 3 (TRS005) had a mean mean of 315.93±94.23mm, P<0.001. 3 It is significantly lower than that of the control group, P<0.05.

[0115] The tumor growth inhibition rates (%TGI) on Day 35 were 96.24% in Group 4, 81.94% in Group 3, and 55.24% in Group 3. The above results indicate that the combination therapy of the anti-CD20 antibody-drug conjugate TRS005 of the present invention and the CHOP regimen significantly inhibited the growth of tumor cells with high CD20 expression, demonstrating significantly improved therapeutic efficacy compared to the R-CHOP regimen and the TRS005 monotherapy. At the same time, the TRS005 monotherapy achieved a more favorable therapeutic effect than the R-CHOP regimen.

[0116] (2) Effect of the test substance on the body weight of tumor-bearing animals The effects of TRS005 monotherapy or combination therapy on the body weight of WSU-DLCL2 tumor-bearing animals showed that on Day 35, the average body weight of animals in each group increased by 1.67% to 8.39% (0.34 to 1.68 g) compared to the start of the experiment on Day 0. The effects of treatment in each group on the body weight of tumor-bearing animals are shown in Table 4.

[0117] Table 4. Effect of administration of each group on the body weight of tumor-bearing animals

[0118] [Table 4]

[0119] During the experimental period, 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 or adverse reactions were observed, demonstrating the good safety profile of the pharmaceutical composition of TRS005 and CHP and the TRS005 monotherapy regimen of the present invention.

[0120] Example 4: Study of the growth inhibitory effect of TRS005 alone and in combination with CHOP compared with R-CHOP regimen on human lymphoma OCI-LY19 cell xenograft tumors

[0121] Drug information, model building, grouping and administration Drug information: As in Example 3, the drug was administered subcutaneously under sterile conditions to the right back at a concentration of 210 mg / kg. 60.1 mL of the OCI-LY19 cell suspension was inoculated into each mouse. The average tumor volume was approximately 150 mm 3 When tumor volume reached 1000 mg / kg, the animals were randomly divided into four groups according to tumor volume: Group 1 vehicle group, Group 2 R-CHOP group, Group 3 TRS005 single agent group, and Group 4 T-CHP group. Each group contained 10 mice, and tumor volume differences were less than 10% of the mean. The day of grouping was designated Day 0, and administration began according to the animals' weight. The specific grouping and administration regimen are shown in Table 5:

[0122] Table 5. Grouping and administration plan

[0123] [Table 5]

[0124] where IV = intravenous injection; IP = intraperitoneal injection; PO = oral administration; BIW x 3w = twice a week for a total of 3 weeks; QW x 3w = once a week for a total of 3 weeks; QD x 5 / w x 3w = once a day, 5 times a week for a total of 3 weeks.

[0125] During the experimental period, animal weights and tumor volumes were measured twice weekly. Animal clinical symptoms were observed and recorded daily. Clinical observations included the animals' overall health, weight and behavioral abnormalities, and other treatment-related adverse reactions. During the experimental period, if an individual experimental animal met any of the following conditions, it was removed from the experimental group and euthanized: 1. The animal's weight decreased by 20% or more compared to Day 0 (BWL ≥ 20%); 2. The animal showed severe adverse reactions, such as blindness or paralysis; or 3. The tumor volume exceeded 2000 mm. 3 Larger; 4, forming an open ulcer on the tumor surface.

[0126] Evaluation indicators Tumor volume (TV) is 1 / 2ab 2 where a and b are the measured length and width of the tumor, respectively.

[0127] The tumor growth inhibition rate (%TGI) was (1-TV T / TV C )×100%, TV C is the mean tumor volume in the negative control group, TV T was calculated based on the mean tumor volume of the treatment group.

[0128] Relative tumor volume (RTV) is V t / V0, where V0 is the tumor volume at the time of grouping, and V t is the tumor volume at each measurement.

[0129] Relative tumor growth rate (%T / C RTV ) is T RTV / C RTV The calculation was done by multiplying the T by 100%. RTV is the treatment group RTV, and C RTV is the negative control group RTV.

[0130] The percent weight change (%BWC) of the animals was calculated using the formula (BWt-BW0) / BW0 x 100%, where BWt is the weight of the animals at each measurement, and BW0 is the weight of the animals at the time of grouping.

[0131] (1) Tumor-inhibiting effect In this experiment, the effects of TRS005 alone or in combination with CHP on the tumor volume of human lymphoma OCI-LY19 are shown in Tables 6-7 and FIG.

[0132] Table 6. Effect of administration of each treatment on tumor volume from D0 to D14

[0133] [Table 6]

[0134] Note: *** indicates P<0.001 compared with vehicle Group 1. ## indicates P<0.01 compared with R-CHOP Group 2. $$$ indicates P<0.001 compared with Group 3.

[0135] On Day 14, the tumor volume of the vehicle control animals was 2000 mm 3 The tumor volume began to exceed 100%, reaching the end point of the study, so the animals were euthanized. Statistical analysis of the data on Day 14 revealed that, as can be seen from Table 6 and Figure 4, the mean tumor volume in Group 4 (T-CHP) on Day 14 was 176.27 ± 63.80 mm. 3 The mean tumor volume in Group 2 (R-CHOP) was 557.11 ± 76.43 mm 3 The mean tumor volume in Group 3 (TRS005) was 905.30 ± 129.66 mm 3 The mean tumor volume in Group 4 (T-CHP) was significantly lower than in Group 2 (R-CHOP) and Group 3 (TRS005). The tumor growth inhibition rates (%TGI) on Day 14 were 90.63%, 51.87%, and 70.38% for Group 4, Group 3, and Group 2, respectively.

[0136] Table 7. Changes in tumor volume in mice in each administration group from D14 to D28

[0137] [Table 7]

[0138] ## indicates P<0.01 compared with group 2 R-CHOP.

[0139] On D14, tumor inhibition rate TGI analysis was performed. However, four mice in Group 1 had tumor volumes exceeding 2000 mm3 on D14 and were euthanized, making it impossible to perform tumor inhibition rate TGI analysis on D14-D28. As can be seen from Table 7, the tumor volumes of Group 4 (T-CHP) mice were significantly lower than those of Group 2 (R-CHOP) during the D14-D24 period, and the tumor inhibition effect continued the trend from D0-D14. On D24, one mouse in Group 4 (T-CHP) and seven mice in Group 2 (R-CHOP), respectively, had tumor volumes exceeding 2000 mm3. 3By day 28, only three mice remained in Group 2 (R-CHOP) that could be used for tumor volume measurement, and the tumor volume was 327.42 mm. 3 , 2458.12mm 3 , 2798.35mm 3 The mean value was 1861.30 ± 773.20. In Group 4 (T-CHP), tumor volume was measured in nine mice, and the tumor volume was 687.44 mm. 3 , 241.60mm 3 , 57.61mm 3 , 281.22mm 3 , 1243.65mm 3 , 0.00mm 3 , 723.03mm 3 , 1810.62mm 3 , 1673.31mm 3 The mean value was 746.50±228.09.

[0140] Combining the tumor volume data of mice in each treatment group from D0 to D28, the tumor volume of Group 4 (T-CHP) mice was significantly lower than that of Group 2 (R-CHOP) and Group 3 (TRS005 monotherapy), and Group 4 (T-CHP) was able to significantly suppress the growth of tumor cells with low CD20 expression, resulting in a significantly improved therapeutic effect compared to Group 2 (R-CHOP) and Group 3 (TRS005 monotherapy).

[0141] (2) Effect of the test substance on the body weight of tumor-bearing animals The effects of TRS005 administration alone or in combination on the body weight of OCI-LY19 tumor-bearing animals showed that on Day 14, the average body weight of animals in each group increased by 1.11-12.50% (0.24-2.64 g) compared to the start of the experiment on Day 0. The effects of treatment in each group on the body weight of tumor-bearing animals are shown in Table 8.

[0142] Table 8. Effect of administration of each group on the body weight of tumor-bearing animals

[0143] [Table 8]

[0144] During the experimental period, 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 toxicities or adverse reactions were observed, demonstrating that the pharmaceutical composition of TRS005 and CHP and the TRS005 monotherapy of the present invention have good safety in inhibiting tumor cells with low CD20 expression.

[0145] Based on the above, the present invention provides a new approach to drug combination therapy based on the existing first-line standard therapy for NHL, particularly DLBCL. By subtracting from the existing first-line standard therapy, R-CHOP, the anti-CD20 antibody-drug conjugates of the present invention can replace rituximab and vincristine (O) in the standard therapy, thereby reducing the number of medications administered. Even with reduced dosages, the pharmaceutical composition of the anti-CD20 antibody-drug conjugates of the present invention and CHOP can achieve significantly improved tumor inhibitory effects in both high- and low-CD20-expressing DLBCL cells compared to R-CHOP. Furthermore, the anti-CD20 antibody-drug conjugates of the present invention alone exhibit excellent therapeutic effects in both high- and low-CD20-expressing DLBCL cells, providing clinical benefits with fewer medications, shorter infusion times, lower safety risks, and better economical outcomes, making them an excellent treatment option for first-line DLBCL treatment.

[0146] The above description of the specific embodiments of the present invention is not intended to limit the present invention, and those skilled in the art may make various modifications or variations based on the present invention without departing from the spirit of the present invention, all of which are within the scope of the claims of the present invention.

Claims

1. 1. Use of an anti-CD20 antibody-drug conjugate, alone or in combination with at least one therapeutic agent, in the manufacture of a therapeutic agent for treating non-Hodgkin's lymphoma, wherein the anti-CD20 antibody-drug conjugate has the following structure: 【Chemistry 1】 ； wherein mAb is a recombinant anti-CD20 monoclonal antibody, preferably the recombinant anti-CD20 monoclonal antibody is rituximab or a biologically similar drug thereof, the drug-antibody ratio (DAR) is n, wherein n is 1 to 8, preferably n is 4.2±1, more preferably n is 4.2±0.5, and even more preferably n is 4.2±0.3; wherein the at least one therapeutic agent is one or more selected from the group consisting of alkylating agents, antibiotics, botanical preparations, hormone-based drugs, platinum preparations, and antimetabolites; the alkylating agent is one or more selected from the group consisting of cyclophosphamide, ifosfamide, melphalan, busulfan, chlormethine, chlorambucil, thiotepa, bendamustine, carmustine, nimustine, lomustine, and semustine; the antibiotic is at least one selected from the group consisting of doxorubicin, daunorubicin, bleomycin, mitomycin, epirubicin, aclarubicin, and idarubicin; the plant component preparation is one or more selected from the group consisting of vinca alkaloids, vinblastine, vincristine, vindesine, vinorelbine, docetaxel, camptothecin, irinotecan, exatecan, topotecan, paclitaxel, and taxanes; the hormone-based drug is one or more selected from the group consisting of prednisone and dexamethasone; the platinum agent is one or more selected from the group consisting of carboplatin, cisplatin, and oxaliplatin; The antimetabolite is one or more selected from the group consisting of pemetrexed, methotrexate, fluorouracil, capecitabine, cytarabine, gemcitabine, 6-mercaptopurine, and thioguanine.

2. 2. The use of claim 1, wherein the at least one therapeutic agent is an alkylating agent, an antibiotic, or a hormonal agent; the alkylating agent is one or more selected from the group consisting of cyclophosphamide, ifosfamide, melphalan, busulfan, chlormethine, chlorambucil, thiotepa, bendamustine, carmustine, nimustine, lomustine, and semustine; the antibiotic is one or more selected from the group consisting of doxorubicin, daunorubicin, bleomycin, mitomycin, epirubicin, aclarubicin, and idarubicin; The hormone-based drug is at least one selected from the group consisting of prednisone and dexamethasone.

3. 3. The use according to claim 2, wherein the at least one therapeutic agent is cyclophosphamide, doxorubicin and prednisone.

4. The use according to any one of claims 1 to 3, characterized in that the non-Hodgkin's lymphoma is selected from the group consisting of small lymphocytic lymphoma, B-cell chronic lymphocytic lymphoma, marginal zone B-cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, mantle cell lymphoma, precursor B-cell lymphoblastic leukemia / lymphoma (B-ALL / LBL), T-cell large granular lymphocytic leukemia, or anaplastic large cell lymphoma (ALCL).

5. 5. The use according to claim 4, wherein the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma, preferably CD20-positive diffuse large B-cell lymphoma.

6. 5. The use according to claim 4, wherein the non-Hodgkin's lymphoma is a newly diagnosed diffuse large B-cell lymphoma, preferably a newly diagnosed CD20-positive diffuse large B-cell lymphoma.

7. The use according to claim 5, wherein the CD20-positive diffuse large B-cell lymphoma is diffuse large B-cell lymphoma with high CD20 expression or low CD20 expression.

8. The use according to claim 6, characterized in that the newly diagnosed CD20-positive diffuse large B-cell lymphoma is a newly diagnosed CD20-high or CD20-low expressing diffuse large B-cell lymphoma.

9. The use according to claim 7, characterized in that the CD20-positive diffuse large B-cell lymphoma is diffuse large B-cell lymphoma with dual expression of MYC and BCL2 and high or low expression of CD20.

10. The use according to claim 8, characterized in that the newly diagnosed CD20-positive diffuse large B-cell lymphoma is a newly diagnosed diffuse large B-cell lymphoma that double expresses MYC and BCL2 and has high or low CD20 expression.

11. 1. A medicament for treating non-Hodgkin's lymphoma, said medicament comprising an anti-CD20 antibody-drug conjugate and at least one therapeutic agent, and characterized in that said anti-CD20 antibody-drug conjugate has the structure: 【Chemistry 2】 ； wherein mAb is a recombinant anti-CD20 monoclonal antibody, preferably the recombinant anti-CD20 monoclonal antibody is rituximab or a biologically similar drug thereof, the drug-antibody ratio (DAR) is n, wherein n is 1 to 8, preferably n is 4.2±1, more preferably n is 4.2±0.5, and even more preferably n is 4.2±0.3; wherein the at least one therapeutic agent is one or more selected from the group consisting of alkylating agents, antibiotics, botanical preparations, hormone-based drugs, platinum preparations, and antimetabolites; the alkylating agent is one or more selected from the group consisting of cyclophosphamide, ifosfamide, melphalan, busulfan, chlormethine, chlorambucil, thiotepa, bendamustine, carmustine, nimustine, lomustine, and semustine; the antibiotic is one or more selected from the group consisting of doxorubicin, daunorubicin, bleomycin, mitomycin, epirubicin, aclarubicin, and idarubicin; the plant component preparation is one or more selected from the group consisting of vinca alkaloids, vinblastine, vincristine, vindesine, vinorelbine, docetaxel, camptothecin, irinotecan, exatecan, topotecan, paclitaxel, and taxanes; the hormone-based drug is one or more selected from the group consisting of prednisone and dexamethasone; the platinum agent is one or more selected from the group consisting of carboplatin, cisplatin, and oxaliplatin; The antimetabolite is one or more selected from the group consisting of pemetrexed, methotrexate, fluorouracil, capecitabine, cytarabine, gemcitabine, 6-mercaptopurine, and thioguanine.

12. 12. The pharmaceutical product of claim 11, wherein the at least one therapeutic agent is an alkylating agent, an antibiotic, and a hormonal agent. the alkylating agent is one or more selected from the group consisting of cyclophosphamide, ifosfamide, melphalan, busulfan, chlormethine, chlorambucil, thiotepa, bendamustine, carmustine, nimustine, lomustine, and semustine; the antibiotic is one or more selected from the group consisting of doxorubicin, daunorubicin, bleomycin, mitomycin, epirubicin, aclarubicin, and idarubicin; The hormone-based drug is at least one selected from the group consisting of prednisone and dexamethasone.

13. 13. The pharmaceutical product of claim 12, wherein the at least one therapeutic agent is cyclophosphamide, doxorubicin, and prednisone.

14. The pharmaceutical product according to any one of claims 11 to 13, characterized in that the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma, preferably CD20-positive diffuse large B-cell lymphoma.

15. The pharmaceutical product of claim 14, characterized in that the non-Hodgkin's lymphoma is newly diagnosed diffuse large B-cell lymphoma, preferably newly diagnosed CD20-positive diffuse large B-cell lymphoma.

16. The pharmaceutical product described in claim 15, characterized in that the newly diagnosed CD20-positive diffuse large B-cell lymphoma is newly diagnosed diffuse large B-cell lymphoma with high CD20 expression or low CD20 expression.

17. The pharmaceutical product described in claim 16, characterized in that the newly diagnosed CD20-positive diffuse large B-cell lymphoma is newly diagnosed diffuse large B-cell lymphoma with dual expression of MYC and BCL2 and high or low expression of CD20.

18. 1. A method for treating non-Hodgkin's lymphoma, comprising administering to a subject an effective amount of an anti-CD20 antibody-drug conjugate, or a combination thereof with at least one therapeutic agent, wherein the anti-CD20 antibody-drug conjugate has the structure: 【Transformation 3】 ； wherein mAb is a recombinant anti-CD20 monoclonal antibody, preferably the recombinant anti-CD20 monoclonal antibody is rituximab or a biologically similar drug thereof, the drug-antibody ratio (DAR) is n, wherein n is 1 to 8, preferably n is 4.2±1, more preferably n is 4.2±0.5, and even more preferably n is 4.2±0.3; wherein the at least one therapeutic agent is one or more selected from the group consisting of alkylating agents, antibiotics, botanical preparations, hormone-based drugs, platinum preparations, and antimetabolites; the alkylating agent is one or more selected from the group consisting of cyclophosphamide, ifosfamide, melphalan, busulfan, chlormethine, chlorambucil, thiotepa, bendamustine, carmustine, nimustine, lomustine, and semustine; the antibiotic is one or more selected from the group consisting of doxorubicin, daunorubicin, bleomycin, mitomycin, epirubicin, aclarubicin, and idarubicin; the plant component preparation is one or more selected from the group consisting of vinca alkaloids, vinblastine, vincristine, vindesine, vinorelbine, docetaxel, camptothecin, irinotecan, exatecan, topotecan, paclitaxel, and taxanes; the hormone-based drug is one or more selected from the group consisting of prednisone and dexamethasone; the platinum agent is one or more selected from the group consisting of carboplatin, cisplatin, and oxaliplatin; The antimetabolite is one or more selected from the group consisting of pemetrexed, methotrexate, fluorouracil, capecitabine, cytarabine, gemcitabine, 6-mercaptopurine, and thioguanine.

19. 20. The method of claim 18, wherein the at least one therapeutic agent is an alkylating agent, an antibiotic, and a hormonal agent. the alkylating agent is one or more selected from the group consisting of cyclophosphamide, ifosfamide, melphalan, busulfan, chlormethine, chlorambucil, thiotepa, bendamustine, carmustine, nimustine, lomustine, and semustine; the antibiotic is one or more selected from the group consisting of doxorubicin, daunorubicin, bleomycin, mitomycin, epirubicin, aclarubicin, and idarubicin; The hormone-based drug is at least one selected from the group consisting of prednisone and dexamethasone.

20. 20. The method of claim 19, wherein the at least one therapeutic agent is cyclophosphamide, doxorubicin, and prednisone.

21. The method of any one of claims 18 to 20, wherein the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma, preferably CD20-positive diffuse large B-cell lymphoma.

22. 22. The method of claim 21, wherein the non-Hodgkin's lymphoma is a newly diagnosed diffuse large B-cell lymphoma, preferably a newly diagnosed CD20-positive diffuse large B-cell lymphoma.

23. The method of claim 22, wherein the newly diagnosed CD20-positive diffuse large B-cell lymphoma is a newly diagnosed diffuse large B-cell lymphoma with high CD20 expression or low CD20 expression.

24. The method of claim 23, wherein the newly diagnosed CD20-positive diffuse large B-cell lymphoma is a newly diagnosed diffuse large B-cell lymphoma that double expresses MYC and BCL2 and has high or low CD20 expression.

25. The method of any one of claims 18 to 24, wherein the administration of the anti-CD20 antibody-drug conjugate can be administered before, simultaneously with and / or after the administration of at least one therapeutic agent.

26. A non-therapeutic method for suppressing non-Hodgkin's lymphoma cells, comprising administering to a non-Hodgkin's lymphoma cell line an effective amount of the pharmaceutical agent according to any one of claims 11 to 17.

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