Methods of treating DLBCL using BTK inhibitors and combinations thereof

Abstract

The present invention provides methods for treating non-GCB DLBCL with (S)-7-(1-acryloylpiperidin-4-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo-[1,5-a]pyrimidine-3-carboxamide or a pharmaceutically acceptable salt thereof. The present invention provides methods for treating non-GCB DLBCL in a subject, comprising administering to a subject in need thereof a combination of (S)-7-(1-acryloylpiperidin-4-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo-[1,5-a]pyrimidine-3-carboxamide or a pharmaceutically acceptable salt thereof and an anti-CD20 antibody.

Description

Technical Field

[0001] The present invention discloses methods for treating non-GCB DLBCL in a subject, comprising administering to a subject in need thereof (S)-7-(1-acryloylpiperidin-4-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo-[1,5-a]pyrimidine-3-carboxamide or a pharmaceutically acceptable salt thereof. The present invention also discloses methods for treating non-GCB DLBCL in a subject, comprising administering to a subject in need thereof (S)-7-(1-acryloylpiperidin-4-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo-[1,5-a]pyrimidine-3-carboxamide or a pharmaceutically acceptable salt thereof in combination with an anti-CD20 antibody. Background Art

[0002] Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin's lymphoma, which is clinically heterogeneous: 40% of patients respond well to existing therapies and have prolonged survival, while the rest die from the disease (Alizadeh et al., Nature 2000; 403(6769): 503-11). DLBCL can be divided into subgroups with germinal center B cell-like (GCB), activated B cell-like (ABC), expression profiles according to their prognostic significance (Alizadeh see above). The GCB group has a better survival rate than the ABC group, but both indications are associated with poor clinical outcomes. Molecular profiles have previously been used to predict survival in DLBCL (Rosenwald et al., N Engl J Med. [New England Journal of Medicine] 2002; 346: 1937-1947). In 2016, the World Health Organization (WHO) revised its classification of lymphoid neoplasms to account for major advances in lymphoma biology since 2008 (Swerdlow et al., Blood. 2016; 127(20): 2375-2390). The WHO classification emphasizes changes in gene expression and clinical importance, such as changes in the MYC, BCL2, and / or BCL6 oncogenes. More than 80% of patients with so-called "double-hit" lymphomas have MYC and BCL2 translocations (Aukema et al., Blood. 2011; 117(8): 2319-2331). This suggests the need for additional molecular markers in addition to MYC and BCL2 that can be used to classify patients before treatment or indicate response to treatment.

[0003] Bruton's tyrosine kinase (BTK) inhibitors alone or in combination with CD20 antibodies have shown only modest activity in R / R non-GCB DLBCL. However, patients with CD79B mutations responded frequently to ibrutinib (5 / 9; 55%), especially those with concomitant myeloid differentiation primary response 88 (MYD88) mutations (4 / 5; 80%). The Phase 3 PHONIX trial of ibrutinib + R-CHOP failed to reach its primary endpoint for the treatment of first-line non-GCB DLBCL patients, however, biomarker analysis showed improved event-free survival in the BCL2 / MYC co-expressing population. And these benefits were only observed in patients under 65 years of age, which also suggests that the combination of ibrutinib and R-CHOP may lead to relatively higher toxicity. Therefore, these clinical results indicate that patient stratification and patient response based on molecular markers will be key to future treatment of non-GCB DLBCL, and further research should be done on less toxic regimens. Summary of the invention

[0004] WO 2014 / 173289 A discloses a series of BTK inhibitors, in particular (S)-7-(1-acryloylpiperidin-4-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide (hereinafter referred to as Compound 1). Compound 1 can be used to treat cancers in which there are aberrations in the B cell receptor (BCR) and FcR signaling pathways in which BTK plays an important role, and has been shown to have effective and irreversible inhibitory activity against BTK.

[0005]

[0006] The present invention describes (S)-7-(1-acryloylpiperidin-4-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 1) alone or a pharmaceutically acceptable salt thereof, and a combination of Compound 1 and an anti-CD20 antibody, which show sensitive responses in some populations of non-GCB DLBCL or ABC DLBCL, particularly those with high expression of specific genes (including PAX5, PIM1, BCL2, FOXP1, BCL2, MYC, or BCL2 / MYC).

[0007] The present invention also describes that Compound 1 or a pharmaceutically acceptable salt thereof, when administered alone or as a combination of Compound 1 and an anti-CD20 antibody, shows a sensitive response in some populations of non-GCB DLBCL, particularly those with specific gene mutations, including mutations in the BCR or NOTCH1 pathways, such as CD79B mutations or NOTCH1 mutations.

[0008] The present inventors found that ABC or non-GCB DLBCLs were more sensitive to compound 1 than the GCB subtype in both in vivo and in vitro assays.

[0009] In clinical studies, the inventors found that PAX5 expression was significantly higher in Compound 1 monotherapy responders, and PIM1, BCL2, and FOXP1 expression was higher in Compound 1 combination therapy responders. In Compound 1 monotherapy, the inventors also found that patients with NOTCH1 mutations had higher ORRs. In both Compound 1 monotherapy and combination therapy of Compound 1 in combination with anti-CD20 antibodies, the inventors also found that patients with MYC, BCL2, MYC and BCL2 double-expressing DLBCL tended to have higher ORRs and longer progression-free survival and overall survival, and patients with CD79B showed significantly higher ORRs than patients without CD79B mutations.

[0010] Furthermore, the present inventors found that EBV-negative non-GCB DLBCLs were more sensitive to Compound 1 than EBV-positive non-GCB DLBCLs.

[0011] In a first aspect, the present invention discloses a method of treating non-GCB DLBCL or ABC DLBCL in a subject, the method comprising administering to the subject in need thereof a therapeutically effective amount of Compound 1 or a pharmaceutically acceptable salt thereof.

[0012] The present invention also discloses a pharmaceutical composition for treating non-GCB DLBCL or ABC DLBCL, wherein the pharmaceutical composition comprises Compound 1 or a pharmaceutically acceptable salt thereof.

[0013] In one embodiment, the non-GCB DLBCL is a non-GCB DLBCL with high expression of PAX5, BCL2, MYC, or BCL2 / MYC genes.

[0014] In one embodiment, the non-GCB DLBCL is a non-GCB DLBCL with a BCR pathway gene mutation or a NOTCH1 pathway gene mutation. In a preferred embodiment, the non-GCB DLBCL is a non-GCB DLBCL with a CD79B mutation or a NOTCH1 mutation.

[0015] In a second aspect, the present invention discloses a method of treating non-GCB DLBCL or ABC DLBCL in a subject, the method comprising administering to the subject in need thereof a therapeutically effective amount of a combination of Compound 1 or a pharmaceutically acceptable salt thereof and an anti-CD20 antibody.

[0016] In yet another aspect, the present invention discloses a combination of Compound 1 or a pharmaceutically acceptable salt thereof and an anti-CD20 antibody for use in treating non-GCB DLBCL or ABC DLBCL.

[0017] The present invention also provides use of a drug combination in the manufacture of a drug for treating non-GCB DLBCL or ABC DLBCL, and the drug combination comprises Compound 1 or a pharmaceutically acceptable salt thereof, and an anti-CD20 antibody.

[0018] An article of manufacture, or "kit" comprises a first container, a second container, and a package insert, wherein the first container comprises at least one dose of a medicament comprising Compound 1 or a pharmaceutically acceptable salt thereof, the second container comprises at least one dose of a medicament comprising an anti-CD20 antibody, and further comprises a package insert comprising instructions for treating non-GCB DLBCL in a subject using the medicament.

[0019] In one embodiment, the anti-CD20 antibody is selected from rituximab, ibritumomab tiuxetan, tositumomab, ofatumumab, or obinutuzumab.

[0020] In one embodiment, the non-GCB DLBCL is a non-GCB DLBCL with high expression of PIM1, BCL2, FOXP1, MYC, or BCL2 / MYC genes.

[0021] In one embodiment, the non-GCB DLBCL is a non-GCB DLBCL with a BCR pathway gene mutation or a NOTCH1 pathway gene mutation. In a preferred embodiment, the non-GCB DLBCL is a non-GCB DLBCL with a CD79B mutation or a NOTCH1 mutation.

[0022] In one embodiment in each of the above aspects, the non-GCB is a R / R non-GCB DLBCL.

[0023] In one embodiment of each of the above aspects, the non-GCB is R / R ABC DLBCL.

[0024] In a third aspect, the present invention discloses a method for increasing a cancer patient's response to treatment with a BTK inhibitor, the method comprising:

[0025] a) administering a BTK inhibitor to the patient;

[0026] b) determining the gene expression of PAX5 in a cancer sample obtained from the patient;

[0027] c) comparing PAX5 gene expression, CD79B mutation or NOTCH1 mutation in a cancer sample obtained from a patient treated with a BTK inhibitor with PAX5 gene expression, CD79B mutation or NOTCH1 mutation in a cancer sample before administration of the BTK inhibitor;

[0028] d) wherein increased PAX5 expression, CD79B mutation or NOTCH1 mutation indicates that the patient will be sensitive to BTK inhibitor treatment.

[0029] In a fourth aspect, the present invention discloses a method for increasing a cancer patient's response to treatment with a BTK inhibitor in combination with an anti-CD20 antibody, the method comprising:

[0030] a) administering to the patient a combination of a BTK inhibitor and an anti-CD20 antibody;

[0031] b) determining the gene expression of any one of PIM1, BCL2, FOXP1, MYC or CD79B mutation in a cancer sample obtained from the patient;

[0032] c) comparing the gene expression of any one of PIM1, BCL2, FOXP1, MYC or CD79B mutation in a cancer sample obtained from a patient receiving a combination therapy of a BTK inhibitor and an anti-CD20 antibody with the gene expression of any one of PIM1, BCL2, FOXP1, MYC or CD79B mutation in the cancer sample before administration of the combination of a BTK inhibitor and an anti-CD20 antibody;

[0033] d) wherein increased PIM1, BCL2, FOXP1, MYC expression or CD79B mutation indicates that the patient will be sensitive to BTK inhibitor treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 It was shown that ABC / non-GCB DLBCLs were more sensitive to compound 1 than the GCB subtype.

[0035] DLBCL PDX models were classified into GCB and non-GCB subtypes and treated with compound 1 as described in the methods. Tumor growth inhibition (TGI) of compound 1 was calculated in each mouse. Each dot represents the average TGI of one model. P < 0.05 (two-tailed t test).

[0036] Figure 2 It was shown that non-GCBDLBCL PDX models with CD79B mutations or CD79B-ITAM domain (amino acids 193-210) mutations were more sensitive to compound 1.

[0037] Figure 2(A): Non-GCB DLBCL PDX models were treated with compound 1, and the TGI of each model was calculated as described in the methods. The models were subdivided into CD79B wild-type (CD79B-WT) and CD79B mutant (CD79B-mut) groups. Each dot represents the average TGI of one model. P < 0.05 (two-tailed t-test).

[0038] Figure 2 (B): Non-GCB DLBCL PDX models were treated with compound 1, and the TGI of each model was calculated as described in the methods. The models were subdivided into CD79B ITAM domain (amino acids 193-210) wild type (CD79B ITAM-WT) and CD79B ITAM domain mutant (CD79B ITAM-mut) groups. Each dot represents the average TGI of a model. P < 0.05 (two-tailed t test).

[0039] Figure 3 It was shown that EBV-negative non-GCB DLBCL PDXs were more sensitive to compound 1.

[0040] Non-GCB DLBCL PDX models were treated with compound 1, and TGI for each mouse was calculated as described in the methods. The models were subdivided into Epstein-Barr virus (EBV) negative (EBV-) and EBV positive (EBV+) groups. Each dot represents the mean TGI of one model. P<0.05 (two-tailed t-test).

[0041] FIG. 4 shows mRNA analysis of 56 non-GCB DLBCL subtypes subdivided by the GEP method (95 genes in total).

[0042] FIG. 4(A) shows that PAX5 is enriched in responders in Compound 1 monotherapy regimens 1 and 2.

[0043] FIG. 4(B) shows that PIM1, FOXP1 and BCL2 are enriched in responders via the combination of Compound 1 and CD20 antibody of Schemes 3 and 4.

[0044] FIG5 shows the best observed response (BOR), progression-free survival (PFS), and overall survival (OS) in 28 BCL2-high and 28 BCL2-low patients (using the median expression value of BCL2 as the threshold to define BCL2-high or BCL2-low).

[0045] FIG6 shows the best observed response (BOR), progression-free survival (PFS), and overall survival (OS) in 28 MYC-high and 28 MYC-low patients (using the median expression value of MYC as the threshold to define MYC-high or MYC-low).

[0046] FIG7 shows the best observed response (BOR), progression-free survival (PFS), and overall survival (OS) in 18 DE patients and 38 non-DE patients (using the median expression value of BCL2 / MYC as the threshold to define DE or non-DE). DETAILED DESCRIPTION

[0047] definition

[0048] Unless specifically defined otherwise in this document, all other technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art.

[0049] As used herein, including the appended claims, singular forms such as "a," "an," and "the" include their corresponding plural referents unless the context clearly dictates otherwise.

[0050] Unless the context clearly indicates otherwise, the term "or" is used to mean and is interchangeable with the term "and / or."

[0051] The terms "administration or administering" and "treating or treatment" herein, when applied to an animal, a human, an experimental subject, a cell, a tissue, an organ, or a biological fluid, mean that an exogenous agent, therapeutic agent, diagnostic agent, or composition contacts the animal, human, subject, cell, tissue, organ, or biological fluid. Treatment of cells encompasses contact of an agent with the cell, and contact of an agent with a fluid, wherein the fluid contacts the cell. The terms "administration" and "treatment" also mean, for example, in vitro and ex vivo treatment of cells by agents, diagnostics, binding compounds, or by other cells. The term "subject" herein includes any organism, preferably an animal, more preferably a mammal (e.g., rats, mice, dogs, cats, rabbits), and most preferably a human. In one aspect, treating any disease or disorder refers to improving the disease or disorder (i.e., slowing down or preventing or alleviating the development of the disease or at least one of its clinical symptoms). In another aspect, "treat, treating, or treatment" refers to alleviating or improving at least one physical parameter, including those that the patient may not be able to discern. In yet another aspect, "treat," "treating," or "treatment" refers to regulating the disease or disorder either physically (e.g., stabilization of discernible symptoms), physiologically (e.g., stabilization of a physical parameter), or both. In yet another aspect, "treat," "treating," or "treatment" refers to preventing or delaying the onset or development or progression of the disease or disorder.

[0052] As used herein, the term "therapeutically effective amount" refers to the amount of a Bcl-2 inhibitor that is sufficient to affect such treatment of a disease, disorder, or symptom when administered to a subject for treatment of the disease, disorder, or at least one clinical symptom of such disease or disorder. The "therapeutically effective amount" may vary depending on factors such as the agent, the disease, disorder, and / or symptoms of the disease or disorder, the severity of the disease, disorder, and / or symptoms of the disease or disorder, the age of the subject to be treated, and / or the weight of the subject to be treated. In any given case, the appropriate amount may be apparent to one skilled in the art or may be determined by routine experimentation. In the context of combination therapy, a "therapeutically effective amount" refers to the total amount of the composition used to effectively treat the disease, disorder, or condition.

[0053] The present invention provides a method for treating non-GCB DLBCL or ABC DLBCL in a subject, the method comprising administering Compound 1 or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0054] The present invention also provides a method for treating non-GCB DLBCL or ABC DLBCL in a subject, the method comprising administering to a subject in need thereof a combination of Compound 1 or a pharmaceutically acceptable salt thereof and an anti-CD20 antibody.

[0055] The present invention also provides a method for increasing the response of a cancer patient to treatment with a BTK inhibitor, the method comprising:

[0056] a) administering a BTK inhibitor to the patient;

[0057] b) determining the expression of the PAX5 gene in a cancer sample obtained from the patient;

[0058] c) comparing PAX5 gene expression in cancer samples obtained from patients treated with a BTK inhibitor with PAX5 gene expression in cancer samples before administration of the BTK inhibitor;

[0059] d) wherein increased PAX5 expression indicates that the patient will be sensitive to BTK inhibitor treatment.

[0060] The present invention also provides a method for increasing the response of a cancer patient to a combination treatment with a BTK inhibitor and an anti-CD20 antibody, the method comprising:

[0061] a) administering to the patient a combination of a BTK inhibitor and an anti-CD20 antibody;

[0062] b) determining the gene expression of any one of PIM1, BCL2, FOXP1 or MYC in a cancer sample obtained from the patient;

[0063] c) comparing the gene expression of any one of PIM1, BCL2, FOXP1 or MYC in a cancer sample obtained from a patient receiving a combination of a BTK inhibitor and an anti-CD20 antibody with the gene expression of any one of PIM1, BCL2, FOXP1 or MYC in the cancer sample before administration of the combination of a BTK inhibitor and an anti-CD20 antibody;

[0064] d) wherein increased expression of PIM1, BCL2, FOXP1 or MYC indicates that the patient will be sensitive to treatment with a BTK inhibitor.

[0065] Anti-CD20 antibody

[0066] "Anti-CD20 antibodies" include, but are not limited to, rituximab, ibritumomab tiuxetan, tositumomab, ofatumumab, or obinutuzumab.

[0067] Treatment

[0068] In one aspect, the invention provides a method of treating GCB DLBCL or ABC DLBCL in a subject.

[0069] In certain aspects, the method comprises administering Compound 1, or a pharmaceutically acceptable salt thereof, to a subject in need thereof.

[0070] In one embodiment, the non-GCB DLBCL is a non-GCB DLBCL with high expression of a gene selected from the group consisting of PAX5, BCL2, MYC, or BCL2 / MYC.

[0071] In one embodiment, the non-GCB DLBCL is a non-GCB DLBCL with a BCR pathway gene mutation or a NOTCH1 pathway gene mutation. In a preferred embodiment, the non-GCB DLBCL is a non-GCB DLBCL with a CD79B mutation or a NOTCH1 mutation.

[0072] In yet another aspect, the present invention discloses a combination of Compound 1 or a pharmaceutically acceptable salt thereof and an anti-CD20 antibody for use in treating non-GCB DLBCL or ABC DLBCL.

[0073] In one embodiment, the non-GCB DLBCL is a non-GCB DLBCL with high expression of a gene selected from the group consisting of PIM1, BCL2, FOXP1, MYC, or BCL2 / MYC.

[0074] In one embodiment, the non-GCB DLBCL is a non-GCB DLBCL with a BCR pathway gene mutation or a NOTCH1 pathway gene mutation. In a preferred embodiment, the non-GCB DLBCL is a non-GCB DLBCL with a CD79B mutation or a NOTCH1 mutation.

[0075] Compound 1 can be administered in any suitable manner, including oral, parenteral, intrapulmonary, and intranasal administration, and if local treatment is required, it can also be administered within the lesion site. Administration can be carried out by any suitable route. Various dosing regimens include, but are not limited to, single or multiple administrations at various time points, and push injection administration and pulse infusion are contemplated herein.

[0076] Compound 1 will be formulated, dosed, and administered in a manner consistent with good medical practice. Factors considered in this context include the specific disorder being treated, the specific mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the administration regimen, and other factors known to practitioners.

[0077] For the prevention or treatment of disease, appropriate dosages of Compound 1 and anti-CD20 antibody in monotherapy or combination therapy will depend on the type of disease to be treated, the severity and course of the disease.

[0078] In some embodiments, Compound 1 is orally administered at a dose of 40 mg-640 mg per day. In some embodiments, Compound 1 is orally administered at a dose of 100 mg-400 mg per day. In some embodiments, Compound 1 is orally administered at a dose of 320 mg QD or 160 mg BID.

[0079] In some embodiments, the anti-CD20 antibody is obinutuzumab. In some embodiments, Compound 1 is administered orally at a dose of 320 mg QD or 160 mg BID over a 28-day cycle, and obinutuzumab is administered at a loading dose of 1000 mg 3 times per week, followed by 1000 mg on the first day of cycles 2-6.

[0080] In some embodiments, the anti-CD20 is rituximab. Compound 1 is administered in combination with rituximab. In some embodiments, Compound 1 is administered orally at a dose of 320 mg QD or 160 mg BID, rituximab is administered intravenously at 375 mg / m2 on days 1, 8, 15, 22 of cycle 1 and day 1 of cycles 4, 6, 8, 10, and Compound 1 is administered at least 30 minutes before the start of rituximab infusion.

[0081] Abbreviations and definitions

[0082]

[0083]

[0084] Examples

[0085] The present invention is further illustrated by the following examples which illustrate the invention but are not limited thereto.

[0086] Example 1 Comparison of the effects of DLBCL origin cells (COO) and compound 1 response

[0087] method

[0088] DLBCL patient-derived xenograft (PDX) models were established in immunodeficient NCG (NOD-Prkdcem26Cd52Il2rgem26Cd22 / NjuCrl) mice. Based on immunohistochemistry (IHC) staining, these models were classified into GCB and non-GCB subgroups using the Hans algorithm (Hans, CP, et al., Blood, 2004.103(1): p.275-282.). Tumor tissue was subcutaneously inoculated in NCG mice for efficacy studies. When the tumor reached 100-200 mm 3 At 14:00, mice were randomized into groups of 7-10 mice / group based on tumor volume. Compound 1 was formulated with 0.5% methylcellulose as a vehicle and treatment was initiated on the day of randomization. Compound 1 was orally administered at 7.5 mg / (kg body weight), BID. Tumor volume was measured twice a week in two dimensions using a caliper and expressed in mm using the following formula: 3 Indicates volume:

[0089] V=0.5(axb 2 ), where a and b are the long and short diameters of the tumor, respectively. Tumor growth inhibition (TGI) was calculated using the following formula:

[0090] %TGI=100×[1-(treatment vt -treat v0 ) / (vehicle vt -vehicle v0 )]

[0091] treat vt = tumor volume of the treatment group at time t.

[0092] treat v0 = tumor volume of the treatment group at the start of treatment.

[0093] vehicle vt = tumor volume of the vehicle group at time t.

[0094] vehicle v0= tumor volume of the vehicle group at the start of treatment.

[0095] P values ​​were calculated using a two-tailed t test.

[0096] result

[0097] Compound 1 resulted in a higher TGI in the non-GCB DLBCL model (TGI 63.60±42.92, mean±SD) than in the GCB subtype (TGI 25.5±11.96, mean±SD), P<0.05 (two-tailed t-test). Figure 1 .

[0098] in conclusion

[0099] DLBCL of the non-GCB subtype were significantly more sensitive to compound 1 than those of the GCB subtype.

[0100] Example 2 Comparison of sensitivity of CD79B and CD79B ITAM domain mutations to compound 1 in DLBCL

[0101] method

[0102] DLBCL patient-derived xenograft (PDX) models were established in immunodeficient NCG (NOD-Prkdcem26Cd52Il2rgem26Cd22 / NjuCrl) mice. Based on immunohistochemistry (IHC) staining, these models were classified into GCB and non-GCB subgroups using the Hans algorithm. Non-GCB DLBCL PDX models were classified into CD79B mutant versus CD79B wild-type groups; or CD79B ITAM domain mutant (amino acids 193-210) versus CD79B ITAM domain wild-type groups. Tumor tissues were subcutaneously inoculated in NCG mice for efficacy studies. When the tumor reached 100-200mm 3 At 14:00, mice were randomized into groups of 7-10 mice / group based on tumor volume. Compound 1 was formulated with 0.5% methylcellulose as a vehicle and treatment was initiated on the day of randomization. Compound 1 was orally administered at 7.5 mg / (kg body weight), BID. Tumor volume was measured twice a week in two dimensions using a caliper and expressed in mm using the following formula: 3 Indicates volume:

[0103] V=0.5(axb 2 ), where a and b are the long and short diameters of the tumor, respectively. Tumor growth inhibition (TGI) was calculated using the following formula:

[0104] %TGI=100×[1-(treatment vt -treat v0 ) / (vehicle vt -vehiclev0 )]

[0105] treat vt = tumor volume of the treatment group at time t.

[0106] treat v0 = tumor volume of the treatment group at the start of treatment.

[0107] vehicle vt = tumor volume of the vehicle group at time t.

[0108] vehicle v0 = tumor volume of the vehicle group at the start of treatment.

[0109] P values ​​were calculated using a two-tailed t test.

[0110] result

[0111] Compound 1 resulted in higher TGI in CD79B mutant (TGI 87.5±33.14, mean±SD) or CD79B ITAM domain mutant (TGI 96.00±28.83, mean±SD) non-GCB DLBCL models than in CD79B wild-type (TGI 49.44±42.36, mean±SD) or CD79B ITAM domain wild-type (TGI 49.18±41.03, mean±SD) non-GCB DLBCL models, P<0.05 (two-tailed t-test). Figure 2 .

[0112] in conclusion

[0113] Non-GCB DLBCLs with CD79B gene mutations were more sensitive to Compound 1 than those with wild-type CD79B gene. Non-GCB DLBCLs with CD79B ITAM domain mutations were more sensitive to Compound 1 than those without CD79B ITAM domain mutations.

[0114] Example 3 Comparison of EBV infection status and sensitivity to compound 1 in DLBCL

[0115] method

[0116] DLBCL patient-derived xenograft (PDX) models were established in immunodeficient NCG (NOD-Prkdcem26Cd52Il2rgem26Cd22 / NjuCrl) mice. Based on immunohistochemistry (IHC) staining, these models were classified into GCB and non-GCB subgroups using the Hans algorithm. Non-GCB DLBCL PDX models were classified into Epstein-Barr virus (EBV) negative (EBV-) and EBV positive (EBV+) subgroups. Tumor tissue was subcutaneously inoculated in NCG mice for efficacy studies. When the tumor reached 100-200mm 3 At 14:00, mice were randomized into groups of 7-10 mice / group based on tumor volume. Compound 1 was formulated with 0.5% methylcellulose as a vehicle and treatment was initiated on the day of randomization. Compound 1 was orally administered at 7.5 mg / (kg body weight), BID. Tumor volume was measured twice a week in two dimensions using a caliper and expressed in mm using the following formula: 3 Indicates volume:

[0117] V=0.5(axb 2 ), where a and b are the long and short diameters of the tumor, respectively. Tumor growth inhibition (TGI) was calculated using the following formula:

[0118] %TGI=100×[1-(treatment vt -treat v0 ) / (vehicle vt -vehicle v0 )]

[0119] treat vt = tumor volume of the treatment group at time t.

[0120] treat v0 = tumor volume of the treatment group at the start of treatment.

[0121] vehicle vt = tumor volume of the vehicle group at time t.

[0122] vehicle v0 = tumor volume of the vehicle group at the start of treatment.

[0123] The mean TGI for each model was used and is represented by dots in the figures. P values ​​were calculated using a two-tailed t-test to compare differences between groups.

[0124] result

[0125] Compound 1 resulted in a higher TGI in the EBV-negative (TGI 78.00±43.41, mean±SD) non-GCB DLBCL model than in the EBV-positive (TGI 33.25±22.41, mean±SD) model, P<0.05 (two-tailed t-test). Figure 3 .

[0126] in conclusion

[0127] EBV-negative non-GCB DLBCLs were more sensitive to compound 1 than EBV-positive non-GCB DLBCLs.

[0128] Example 4 Treatment of non-germinal center B-cell-like diffuse large B-cell lymphoma (non-GCBDLBCL) treated with BTK inhibitors

[0129] A total of 121 R / R non-GCB DLBCL patients were recruited in four compound 1 treatment studies conducted in similar time periods. In the four studies, two were compound 1 monotherapy (n=79) and two were compound 1 combined with anti-CD20 antibodies (n=42). Similar inclusion and exclusion criteria and response evaluation criteria were used in all studies. Fifty-six non-GCB patients were further subdivided using the HTG EdgeSeq DLBCL origin cell assay based on gene expression profiles (GEP). The expression of approximately 90 lymphoma-related genes from the HTG GEP assay was analyzed by the R limma package to analyze its association with the response to compound 1 treatment. Seventy-seven patient samples were tested by next-generation sequencing (NGS) with a set of genes. The correlation between mutations and objective response rate (ORR) was assessed using the chi-square test.

[0130] Example 4A: Treatment of non-GCB DLBCL with BTK inhibitor monotherapy

[0131] method

[0132] Monotherapy regimen 1: Compound 1 was orally administered daily at 160 mg twice daily (BID) to R / R non-GCB DLBCL patients over a 28-day cycle.

[0133] Monotherapy regimen 2: Compound 1 was orally administered daily at 160 mg BID to R / R non-GCBD BCL patients in each cycle of 28 days.

[0134] 79 non-GCB DLBCL patients received compound 1 monotherapy (monotherapy regimen 1 or 2).

[0135] result

[0136] The unadjusted ORRs in non-GCB DLBCL were 31.6% and 29.3% for the two monotherapies, and 53.8% and 36%, respectively, for patients with GEP-confirmed activated B-cell DLBCL.

[0137] For the 44 non-GCB patients with HTG gene expression profiles, PAX5 expression was significantly higher in both monotherapy responders (see Figure 4).

[0138] For the 44 non-GCB DLBCL patients who underwent group sequencing, mutations in genes related to the B cell receptor (BCR) pathway or NOTCH1 pathway have been identified to be associated with better responses. Non-GCB DLBCL patients with CD79B mutations (n=17) showed a higher ORR than other populations (52.9% vs. 33.3%, p=0.2). In both monotherapy, patients with NOTCH1 mutants (n=3) had a higher ORR (100% vs. 36.6%, p<0.05). See Table 1.

[0139] Table 1

[0140]

[0141]

[0142] [1] 95% CI for the difference based on the Miettinen and Nuriminen method

[0143] [2] Chi-square test

[0144] in conclusion

[0145] In monotherapy treatment regimens, patients with higher PAX5 expression or NOTCH1 or CD79B mutations showed better responses after administration of Compound 1. Likewise, screening of non-GCB DLBCL patients for PAX5 expression or NOTCH1 or CD79B mutations prior to treatment is a useful marker to determine a patient's response to Compound 1 and to determine whether Compound 1 treatment should be continued.

[0146] Example 4B: Treatment of non-GCB DLBCL with BTK inhibitors in combination with anti-CD20 antibodies

[0147] method

[0148] Combination therapy regimen 1:Compound 1 was orally administered daily in combination with obinutuzumab to patients with R / R non-GCBDLBCL at 160 mg BID in each cycle of 28-day cycles. Obinutuzumab was administered for up to 6 cycles in accordance with the US labeling regimen (100 mg on day 1 of cycle 1, 1000 mg on days 8 and 15, 900 mg on day 2 of cycle 1, and 1000 mg on day 1 of subsequent cycles 2-6).

[0149] Combination therapy regimen 2: Compound 1 was orally administered daily in combination with rituximab to R / R non-GCBDLBCL patients at 160 mg BID in each cycle of 28-day cycles. Rituximab was administered intravenously at 375 mg / m2 on days 1, 8, 15, 22 of cycle 1 and day 1 of cycles 4, 6, 8, 10. Compound 1 was administered at least 30 minutes before the start of rituximab infusion.

[0150] Forty-two patients with non-GCB DLBCL received Compound 1 + anti-CD20 antibody combination therapy regimen 1 or 2.

[0151] result

[0152] The unadjusted ORRs in non-GCB DLBCL were 22.7% and 35% for the two combinations, and 50% and 40%, respectively, for patients with GEP-confirmed activated B-cell DLBCL.

[0153] The expression of approximately 90 lymphoma-related genes from the HTG GEP assay was analyzed by the R limma package for association with response to combination therapy regimens 1 to 2. For the 12 non-GCB patients with HTG gene expression profiles, PIM1, BCL2, and FOXP1 were expressed more highly in combination therapy responders. (See Figure 4).

[0154] Non-GCB DLBCL patients with CD79B mutation (n=8) showed significantly higher ORR than patients without CD79B mutation (see Table 2).

[0155] Table 2

[0156]

[0157]

[0158] [1] 95% CI for the difference based on the Miettinen and Nuriminen method

[0159] [2] Chi-square test

[0160] in conclusion

[0161] The combination therapy results are very different from the monotherapy results. The enrichment of PAX5 expression after monotherapy is not obvious. On the contrary, increased PIM1, BCL2, and FOXP1 expression were observed in combination therapy patients. Patients with CD79B mutations showed significantly better responses to combination therapy. Therefore, screening for PIM1, BCL2, or FOXP1 expression or CD79B mutations in non-GCB DLBCL patients before treatment is a useful marker to determine the patient's response to compound 1 and anti-CD20 antibodies and to determine whether combination therapy should be continued.

[0162] Example 4C: Treatment of non-GCB DLBCL with BTK inhibitors monotherapy or combination therapy

[0163] method

[0164] Monotherapy regimen 1: Compound 1 was orally administered daily at 160 mg twice daily (BID) to R / R non-GCB DLBCL patients over a 28-day cycle.

[0165] Monotherapy regimen 2: Compound 1 was orally administered daily at 160 mg BID to R / R non-GCBD BCL patients in each cycle of 28 days.

[0166] Combination therapy regimen 1: Compound 1 was orally administered daily in combination with obinutuzumab to patients with R / R non-GCBDLBCL at 160 mg BID in each cycle of 28-day cycles. Obinutuzumab was administered for up to 6 cycles in accordance with the US labeling regimen (100 mg on day 1 of cycle 1, 1000 mg on days 8 and 15, 900 mg on day 2 of cycle 1, and 1000 mg on day 1 of subsequent cycles 2-6).

[0167] Combination therapy regimen 2: Compound 1 was orally administered daily in combination with rituximab to R / R non-GCBDLBCL patients at 160 mg BID in each cycle of 28 days. Rituximab was administered intravenously at 375 mg / m2 on days 1, 8, 15, 22 of cycle 1 and day 1 of cycles 4, 6, 8, 10. Compound 1 was administered at least 30 minutes before the start of rituximab infusion.

[0168] A total of 121 patients with non-GCB DLBCL were included in the response evaluation. 79 patients received compound 1 monotherapy (monotherapy regimen 1 or 2) and 42 patients received compound 1 + anti-CD20 antibody combination therapy ( Combination therapy regimen 1 or 2 ).

[0169] Fifty-six non-GCB patients were further subdivided based on gene expression profiling (GEP) using the HTG EdgeSeq DLBCL cell-of-origin assay. The expression of approximately 90 lymphoma-related genes from the HTG GEP assay was analyzed by the R limma package for association with response to zanubrutinib treatment.

[0170] result

[0171] The median expression value of BCL2 was used as the threshold to define high BCL2 and low BCL2 groups. A total of 28 high BCL2 (BCL2-H) and 28 low BCL2 (BCL2-L) patients were studied. Patients with high BCL2 expression had significantly better BOR (16 / 28, 57% vs. 6 / 28, 21%, p=0.028) and longer PFS (5.3m vs. 2.8m, p=0.03) / OS (10m vs. 6m, p=0.21). See Figure 5A , 5B and 5C.

[0172] The median expression value of MYC was used as the threshold to define high MYC and low MYC groups. A total of 28 high MYC and 28 low MYC patients were studied. Patients with high MYC expression tended to have better BOR (13 / 28, 46% vs. 9 / 28, 32%, p=0.14) and longer PFS (5.4m vs. 2.9m, p=0.52) / OS (10m vs. 7m, p=0.48). See Fig. 6A , 6B and 6C.

[0173] The median expression value of BCL2 / MYC was used as the threshold to define dual expression (DE) or non-DE. Patients (18 DE and 38 non-DE) received two monotherapies and two combination therapies. Patients with DLBCL that dually expressed MYC and BCL2 tended to have a higher ORR (11 / 18, 61% vs. 11 / 38, 29%; P = 0.12) and longer progression-free survival (5.4 months vs. 3.6 months; P = 0.16) and overall survival (10 months vs. 7 months, P = 0.32). See Fig. 7A , 7B and 7C.

[0174] in conclusion

[0175] Patients with MYC, BCL2, or BCL2 / MYC dual-expressing DLBCL tended to have higher ORR and longer progression-free survival and overall survival, with higher response rates in the four Compound 1 treatment studies. Therefore, expression screening of MYC, BCL2, or BCL2 / MYC dual expression in patients with non-GCB DLBCL before treatment is a useful marker to determine the patient's response to Compound 1 and Compound 1 plus anti-CD20 antibody and to determine whether treatment should continue as monotherapy or combination therapy.

[0176] The foregoing examples and descriptions of certain embodiments should be considered as illustrative, rather than limiting, of the present invention as defined in the claims. As will be readily appreciated, many variations and combinations of the above features may be used without departing from the present invention as set forth in the claims. All such variations are intended to be included within the scope of the present invention. All cited references are incorporated herein by reference in their entirety.

Claims

1. Use of a BTK inhibitor in the preparation of a medicament for treating non-GCB DLBCL in a subject, wherein the BTK inhibitor is compound 1 ， or a pharmaceutically acceptable salt thereof; The subject has a CD79B mutation.

2. The use according to claim 1, wherein compound 1 is administered at 320 mg once daily.

3. The use according to claim 1, wherein Compound 1 is administered at 160 mg twice daily.

4. The use according to claim 2 or 3, wherein compound 1 is administered within a 28-day cycle.

5. Use of a combination of a BTK inhibitor and an anti-CD20 antibody in the preparation of a medicament for treating non-GCB DLBCL in a subject, wherein the BTK inhibitor is compound 1 ， or a pharmaceutically acceptable salt thereof; The subject has a CD79B mutation, and the anti-CD20 antibody is rituximab or obinutuzumab.

6. The use according to claim 5, wherein Compound 1 is administered at 320 mg once daily.

7. The use according to claim 5, wherein Compound 1 is administered at 160 mg twice daily.

8. The use according to claim 6 or 7, wherein compound 1 is administered within a 28-day cycle.

9. The use according to claim 5, wherein the anti-CD20 antibody is rituximab.

10. The use according to claim 5, wherein the anti-CD20 antibody is obinutuzumab.

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