Dosing regimens for treating PI3K-related conditions

Abstract

The present application provides methods of treating PI3K delta related conditions using pyrazolopyrimidine derivatives.

Description

[0001] This application is a divisional application of Chinese patent application No. 2019800489184 (application date: May 30, 2019, invention name: Dosage regimen for treating PI3K-related diseases). Technical Field

[0002] The present application provides methods for treating PI3Kδ-related disorders using pyrazolopyrimidine derivatives. Background Art

[0003] Constitutive signaling through the B cell receptor (BCR) plays a key role in the pathogenesis of human B cell malignancies (Gururajan M. et al., The Journal of Immunology, 2006; 176(10):5715-5719) and leads to downstream activation of class I phosphatidylinositol 3-kinases (PI3Ks) (Benschop RJ, Cambier JC, Current Opinion in Immunology, 1999; 11(2):143-151; So L., Fruman DA The Biochemical Journal, 2012; 442(3):465-481). Class I PI3Ks are heterodimeric lipid kinases composed of regulatory (p85 or p101) and catalytic (p110) subunits (Chalhoub N. et al., Annual review of pathology, 2009; 4:127-150). The four tissue-specific p110 subunit isoforms (class IA: α, β, and δ; class IB: γ) each confer a unique physiological function to the corresponding PI3K isoform, including insulin signaling and angiogenesis (PI3Kα) (Foukas LC et al., Nature, 2006; 441:366; Knight ZA et al., Cell, 2006; 125(4):733-747; and Graupera M. et al., Nature, 2008; 453:662), platelet function and thrombosis (PI3Kβ) (Jackson SP et al., Nature Medicine, 2005; 11:507), and leukocyte function and inflammation (PI3Kγ) (Hirsch E. et al., Thrombosis and Haemostasis, 2006; 95(1):29-35). The PI3Kδ isoform serves as a key node in the signaling network that regulates B cell growth and survival, and its abnormal activation is a key event in the malignant transformation of B cells (Kang S. et al., Proceedings of the National Academy of Sciences of the United States of America, 2005; 102(3):802-807; Puri KD, Gold MR, Frontiers in Immunology, 2012; 3:256).Substantial interconnections exist between the BCR and PI3Kδ-mediated signaling networks and other networks important for regulating B cell survival and proliferation, including the Janus kinase signal transducer and activator of transcription (JAK / STAT) pathway (Lam LT et al., Blood, 2008; 111(7):3701-3713; Rui L. et al., Proceedings of the National Academy of Sciences, 2016; 113(46):E7260-E7267), suggesting potential synergistic or synergistic therapeutic effects in B cell malignancies.

[0004] The prognosis of patients with relapsed or refractory B-cell non-Hodgkin lymphoma (NHL) is generally poor. For example, the 5-year overall survival (OS) rate of patients with relapsed follicular lymphoma (FL) (the most common indolent NHL subtype) is only 50% (Casulo C. et al., Journal of Clinical Oncology, 2015; 33 (23): 2516-2522). The prognosis of patients with relapsed aggressive NHL subtypes is even worse, with the median survival of patients with relapsed diffuse large B-cell lymphoma (DLBCL) who have failed first-line and second-line rescue regimens being 3.6 months and 4.4 months, respectively (Van DenNeste E. et al., Bone Marrow Transplant, 2016; 51 (1): 51-57). Current guidelines for the treatment of relapsed B-cell NHL vary by subtype and include chemo- or radioimmunotherapy, targeted therapy using small molecule kinase inhibitors, or immunomodulatory therapy (NCCN Clinical Practice Guidelines in Oncology (NCCN ). B-cell Lymphomas. 7th edition. 2017; 2017; Tilly H. et al., Annals of Oncology, 2015; 26(Suppl_5): v116-v125; Dreyling M. et al., Annals of Oncology, 2016; 27(Suppl_5): v83-v90; Dreyling M. et al., Annals of Oncology, 2017; 28(Suppl_4): iv62-iv71). In addition to systemic therapy, autologous or allogeneic stem cell transplantation (SCT) is also commonly used to treat patients with relapsed B-cell NHL and allogeneic SCT is considered the only treatment option for these patients (Wudhikarn K. et al. Biology of Blood and Marrow Transplantation, 2011; 17(10): 1497-1504; Avivi I. et al., British Journal of Haematology, 2009; 147(5): 719-728; Klyuchnikov E. et al., Bone Marrow Transplant, 2014; 49(1): 1-7; Hamadani M. et al., Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation. 2009; 15(5): 547-553).

[0005] The differential tissue distribution of PI3K isoform factors leads to their different biological functions. Genetic ablation of PI3Kα or PI3Kβ causes embryonic death, indicating that PI3Kα and PI3Kβ have essential and non-redundant functions at least during development (Vanhaesebroeck et al., 2005). In contrast, mice lacking PI3Kγ and PI3Kδ are viable, fertile and have normal life spans, although they show altered immune systems. PI3Kγ lacks macrophages and neutrophils to weaken the recruitment of inflammatory sites and weaken the activation of T cells (Sasaki et al., Science, 2000, 287 (5455): 1040-6). PI3Kδ mutant mice have specific defects in B cell signaling, resulting in impaired B cell development and reduced antibody response after antigen stimulation (Clayton et al., J Exp Med. 2002, 196(6):753-63; Jou et al., Mol Cell Biol. 2002, 22(24):8580-91; Okkenhaug et al., Science, 2002, 297(5583):1031-4).

[0006] The phenotype of PI3Kγ and PI3Kδ mutant mice suggests that these enzymes play a role in inflammation and other immune-based diseases and this has been confirmed in preclinical models. PI3Kγ mutant mice are essentially protected from disease in mouse models of rheumatoid arthritis (RA) and asthma (Camps et al., Nat Med. 2005, 11 (9): 936-43; Thomas et al., Eur J Immunol. 2005, 35 (4): 1283-91). In addition, it is shown that treatment of wild-type mice with a selective inhibitor of PI3Kγ reduces glomerulonephritis and prolongs survival in the MRL-lpr model of systemic lupus nephritis (SLE) and inhibits joint inflammation and damage in the RA model (Barber et al., Nat Med. 2005, 11 (9): 933-5; Camps et al., 2005). Similarly, PI3Kδ mutant mice and wild-type mice treated with a selective inhibitor of PI3Kδ have been shown to have reduced allergic airway inflammation and hyperresponsiveness in a mouse asthma model (Ali et al., Nature. 2004, 431(7011):1007-11; Lee et al., FASEB J. 2006, 20(3):455-65) and reduced disease in an RA model (Randis et al., Eur. J. Immunol., 2008, 38(5):1215-24).

[0007] B cell proliferation has been shown to play an important role in the development of inflammatory autoimmune diseases (Puri, Frontiers in Immunology (2012), 3 (256), 1-16; Walsh, Kidney International (2007) 72, 676–682). For example, B cells support T cell autoreactivity, which is an important component of inflammatory autoimmune diseases. Once activated and mature, B cells can be trafficked to sites of inflammation and recruit inflammatory cells or differentiate into plasmablasts. Therefore, the activity of B cells can be affected by targeting B cell stimulatory cytokines, B cell surface receptors, or by B cell depletion. Rituximab - an IgG1κ mouse / human chimeric monoclonal antibody directed against the B cell surface receptor CD20 - has been shown to deplete CD20+ B cells. The use of rituximab has been shown to be effective in treating idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, or vasculitis. For example, treatment with rituximab results in disease remission in patients with antineutrophil cytoplasmic antibody-associated (ANCA) systemic vasculitis (AASV), suggesting peripheral B cell depletion (Walsh, 2007; Lovric, Nephrol Dial Transplant (2009) 24: 179–185). Similarly, complete responses have been reported in one-third to two-thirds of patients with mixed cryoglobulinemia vasculitis following treatment with rituximab, including patients with severe forms of vasculitis that are resistant or intolerant to other treatments (Cacoub, Ann Rheum Dis 2008; 67: 283–287). Similarly, rituximab has been shown to be effective in treating patients with idiopathic thrombocytopenic purpura (or immune thrombocytopenic purpura) (Garvey, British Journal of Haematology, (2008) 141, 149-169; Godeau, Blood (2008), 112(4), 999-1004; Medeo, European Journal of Haematology, (2008) 81, 165-169) and autoimmune hemolytic anemia (Garvey, British Journal of Haematology, (2008) 141, 149-169).

[0008] PI3Kδ signaling is dependent on B cell survival, migration and activation (Puri, Frontiers in Immunology, 2012, 3(256), 1-16, pp. 1-5; and Clayton, J Exp Med, 2002, 196(6): 753-63). For example, PI3Kδ is required for antigen-dependent B cell activation driven by the B cell receptor. By blocking B cell adhesion, survival, activation and proliferation, PI3Kδ inhibition can weaken the ability of B cells to activate T cells, thereby preventing their activation and reducing the secretion of autoantibodies and proinflammatory cytokines. Therefore, through its ability to inhibit B cell activation, PI3Kδ inhibitors are expected to treat B cell-mediated diseases that can be treated with similar methods such as depletion of B cells by rituximab. In fact, PI3Kδ inhibitors have been shown to be useful in mouse models of various autoimmune diseases that can also be treated with rituximab, such as arthritis (Puri (2012)). Furthermore, innate-like B cells, which are implicated in autoimmunity, are sensitive to PI3Kδ activity, as MZ and B-1 cells are almost absent in mice lacking the p110δ gene (Puri (2012). PI3Kδ inhibitors can reduce the trafficking and activation of MZ and B-1 cells, which are implicated in autoimmune diseases.

[0009] In addition to their potential role in inflammatory diseases, all four class I PI3K isoforms can play a role in cancer. The gene encoding p110α is often mutated in common cancers including breast, prostate, colon and endometrium (Samuels et al., Science, 2004, 304 (5670): 554; Samuels et al., Curr Opin Oncol. 2006, 18 (1): 77-82). 80% of these mutations are represented by one of the three amino acid substitutions in the helical or kinase domains of the enzyme and cause a significant increase in kinase activity, resulting in oncogenic transformation in cell culture and animal models (Kang et al., Proc Natl Acad Sci US A. 2005, 102 (3): 802-7; Bader et al., Proc Natl Acad Sci US A. 2006, 103 (5): 1475-9). Such mutations have not been identified in other PI3K isoforms, although there is evidence that they can promote the development and progression of malignant tumors. Consistent overexpression of PI3Kδ was observed in acute myeloblastic leukemia (Sujobert et al., Blood, 2005, 106 (3): 1063-6) and inhibitors of PI3Kδ can prevent the growth of leukemic cells (Billottet et al., Oncogene. 2006, 25 (50): 6648-59). Increased expression of PI3Kγ is seen in chronic myeloid leukemia (Hickey et al., J Biol Chem. 2006, 281 (5): 2441-50). Alterations in the expression of PI3Kβ, PI3Kγ, and PI3Kδ have also been observed in cancers of the brain, colon, and bladder (Benistant et al., Oncogene, 2000, 19(44):5083-90; Mizoguchi et al., Brain Pathol. 2004, 14(4):372-7; Knobbe et al., Neuropathol Appl Neurobiol. 2005, 31(5):486-90). In addition, these isoforms have been shown to be oncogenic in cell culture (Kang et al., 2006).

[0010] The introduction of rituximab improved the treatment outcomes of patients with B-cell NHL (Chao MP, Cancer Management and Research. 2013; 5: 251-269; Nandagopal L., Mehta A., Expert Review of Hematology, 2017; 10(3): 259-273). PI3K inhibitors have shown promise for patients experiencing rituximab resistance and / or relapsed or refractory disease, but clinical use is limited by toxicity. For example, the first class PI3Kδ selective inhibitor idelalisib is associated with considerable toxicity, including hepatotoxicity (Coutré S.E. et al., Leukemia & Lymphoma, 2015; 56(10): 2779-2786; Gopal A., Graf S., Expert opinion on pharmacotherapy, 2016; 17(2): 265-274) and an increased risk of opportunistic infections (Lampson BL et al., Blood, 2016; 128(2): 195-203), and requires frequent monitoring. Similarly, therapies targeting other PI3K isoforms have corresponding risks (e.g., hyperglycemia and hypertension) (Burris HA et al., Journal of Clinical Oncology, 2014; 32(15_Suppl): 2513-2513; ALIQOPA TM .(copanlisib) for injection, for intravenous use. US Whippany, NJ: Bayer HealthCare Pharmaceuticals Inc.; 2017; Patnaik A. et al., Annals of Oncology, 2016; 27(10): 1928-1940; Dreyling M. et al., Journal of Clinical Oncology, 2017; 35(35): 3898-3905). Therefore, there remains an unmet need for effective therapies with an improved safety profile in this difficult-to-treat patient population. Summary of the invention

[0011] The present application particularly provides a method for treating a disease in a patient, wherein the disease is associated with abnormal expression or activity of PI3Kδ kinase, the method comprising:

[0012] i) administering to the patient a first dose of about 3 mg / day to about 50 mg / day of a PI3Kδ inhibitor for a first period of about 2 weeks to about 12 weeks; and

[0013] ii) administering to the patient a second dose of the PI3Kδ inhibitor at the end of the first period, the second dose being less than the first dose and being:

[0014] (a) about 2.5 mg / day or less; or

[0015] (b) about 50 mg / week or less;

[0016] and wherein the second dose is administered during a second period occurring after the first period.

[0017] The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1A-1B The PK of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride (ie, Compound 1C) at steady state is shown. Figure 1A : PK of compound 1C administration. Figure 1B : Dose-response relationship of compound 1C AUC. AUC: area under the curve; AUC 0–T : Area under the plasma concentration-time curve; IC 50 : half maximal inhibitory concentration; IC 90 : 90% maximum inhibitory concentration; PK: pharmacokinetics; QD: once daily.

[0019] Figure 2 The monotherapy best percent change from baseline in target lesion size is shown. *Data for 3 patients with DLBCL, 2 patients with MZL, and 1 patient with MCL are not shown due to unmeasurable disease at baseline or invalid post-baseline target lesion measurements. Best percent change from baseline in target lesion size >100%. IB: 3 patients with MCL had received ibrutinib prior to the study, with 2 patients achieving a best overall response of CR or PR. DLBCL: diffuse large B-cell lymphoma; FL: follicular lymphoma; MCL: mantle cell lymphoma; MZL: marginal zone lymphoma.

[0020] Figure 3Duration of Compound 1C monotherapy treatment is shown. AE: adverse event; DLBCL: diffuse large B-cell lymphoma; FL: follicular lymphoma; MCL: mantle cell lymphoma; MZL: marginal zone lymphoma; PD: progressive disease; QW: once weekly.

[0021] Figure 4 Mean percent inhibition of phosphorylated AKT at steady state is shown.Compound 1C was administered once daily and on day 15 of cycle 1, levels of phosphorylated AKT (pAKT; Ser473) were measured by flow cytometry in SU-DHL cells added to patient whole blood following Compound 1C treatment.

[0022] Figure 5A-5B ALT ( Figure 5A ) and AST( Figure 5B ). ALT: alanine aminotransferase; AST: aspartate aminotransferase; QD: once daily; ULN: upper limit of normal.

[0023] Figure 6 Compound 1C monotherapy times to achieve responses in NHL subtypes of interest (DLBCL, FL, MCL, MZL) are shown. DLBCL: diffuse large B-cell lymphoma; FL: follicular lymphoma; MCL: mantle cell lymphoma; MZL: marginal zone lymphoma.

[0024] Figure 7 Representative positron emission tomography (PET) images of patients achieving complete response to compound 1C monotherapy are shown. Patients <65 years old, with mantle cell lymphoma without bone marrow involvement (target lesion, 98 mm×58 mm) and have received 1 prior treatment (R-HyperCVAD). Treated with compound 1C, 30 mg, once daily; complete response was achieved at week 9. DETAILED DESCRIPTION

[0025] The present application provides a method for treating a disease in a patient, wherein the disease is associated with abnormal expression or activity of PI3Kδ kinase, the method comprising:

[0026] i) administering to the patient a first dose of about 3 mg / day to about 50 mg / day of a PI3Kδ inhibitor for a first period of about 2 weeks to about 12 weeks; and

[0027] ii) administering to the patient a second dose of the PI3Kδ inhibitor at the end of the first period, the second dose being less than the first dose and being:

[0028] (a) about 2.5 mg / day or less; or

[0029] (b) about 50 mg / week or less;

[0030] and wherein the second dose is administered during a second period occurring after the first period.

[0031] The present application also provides a method for treating a disease in a patient, wherein the disease is associated with abnormal expression or activity of PI3Kδ kinase, the method comprising:

[0032] i) administering to the patient a first dose of about 3 mg / day to about 50 mg / day of a PI3Kδ inhibitor for a first period of about 2 weeks to about 12 weeks; and

[0033] ii) administering to the patient a second dose of the PI3Kδ inhibitor at the end of the first period, the second dose being less than the first dose and being about 7.5 mg / day or less;

[0034] and wherein the second dose is administered during a second period occurring after the first period.

[0035] As used herein, the term "first period" refers to the period during which the first dose of the PI3Kδ inhibitor is administered to the patient.

[0036] In some embodiments, the first period begins when the first dose of the PI3Kδ inhibitor is initially administered to the patient.

[0037] As used herein, the term "second period" refers to the period during which a second dose of a PI3Kδ inhibitor is administered to the patient.

[0038] In some embodiments, the second period begins when the second dose of the PI3Kδ inhibitor is initially administered to the patient.

[0039] In some embodiments, the second dose of the PI3Kδ inhibitor is initially administered to the patient (i.e., the start of the second period) within 2 weeks of the end of the first period, such as within 1 week, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day of the end of the first period.

[0040] In some embodiments, the second period lasts for any length of time, such as until treatment ends.

[0041] In some embodiments, the first doses are each administered as a single, once-daily dose.

[0042] In some embodiments, the first doses are each administered as a single, once-daily oral dose.

[0043] In some embodiments, the first dose is about 5 mg / day to about 50 mg / day, for example, about 5 mg / day to about 40 mg / day, about 5 mg / day to about 30 mg / day, about 5 mg / day to about 20 mg / day, about 5 mg / day to about 10 mg / day, about 10 mg / day to about 40 mg / day, about 10 mg / day to about 30 mg / day, about 10 mg / day to about 20 mg / day, about 20 mg / day to about 40 mg / day, about 20 mg / day to about 30 mg / day, or about 20 mg / day to about 40 mg / day.

[0044] In some embodiments, the first dose is about 50 mg / day, about 45 mg / day, about 40 mg / day, about 30 mg / day, about 20 mg / day, about 15 mg / day, about 10 mg / day, or about 5 mg / day.

[0045] In some embodiments, the first dose is about 18 mg / day to about 22 mg / day.

[0046] In some embodiments, the first dose is about 18 mg / day to about 22 mg / day and is administered as a single, once-daily (ie, QD) dose.

[0047] In some embodiments, the first dose is about 18 mg / day to about 22 mg / day and is administered as a single, once-daily (ie, QD) oral dose.

[0048] In some embodiments, the first dose is about 20 mg / day.

[0049] In some embodiments, the first dose is about 20 mg / day and is administered as a single, once-daily (ie, QD) dose.

[0050] In some embodiments, the first dose is about 20 mg / day and is administered as a single, once-daily (ie, QD) oral dose.

[0051] In some embodiments, the first dose is about 10 mg / day.

[0052] In some embodiments, the first dose is about 10 mg / day and is administered as a single, once-daily (ie, QD) dose.

[0053] In some embodiments, the first dose is about 10 mg / day and is administered as a single, once-daily (ie, QD) oral dose.

[0054] In some embodiments, the first period of time is about 3 weeks to about 11 weeks, such as about 8 weeks to about 12 weeks, about 4 weeks to about 10 weeks, about 5 weeks to about 9 weeks, or about 8 weeks to about 9 weeks. In some embodiments, the first period of time is about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, or about 12 weeks.

[0055] In some embodiments, the first period of time is from about 8 weeks to about 9 weeks.

[0056] In some embodiments, the first period is about 8 weeks.

[0057] In some embodiments, the first period is about 9 weeks.

[0058] In some embodiments, the first dose is reduced during the first period of time.

[0059] In some embodiments, the first dose is reduced from about 3 mg / day to about 50 mg / day to about 3 mg / day to about 30 mg / day, about 3 mg / day to about 20 mg / day, about 3 mg / day to about 15 mg / day, about 3 mg / day to about 10 mg / day, about 3 mg / day to about 7 mg / day, or about 3 mg / day to about 5 mg / day.

[0060] In some embodiments, the first dose is reduced from about 3 mg / day to about 50 mg / day to about 3 mg / day to about 7 mg / day during the first period of time.

[0061] In some embodiments, the first dose is reduced from about 50 mg / day to about 30 mg / day, about 20 mg / day, about 15 mg / day, about 10 mg / day, about 5 mg / day, about 2 mg / day, or about 1 mg / day during the first period.

[0062] In some embodiments, the first dose is reduced from about 20 mg / day to about 15 mg / day, about 10 mg / day, about 5 mg / day, about 2 mg / day, or about 1 mg / day during the first period.

[0063] In some embodiments, the second doses are each administered as a single, once-daily (QD) dose or as a single, once-weekly (QW) dose.

[0064] In some embodiments, the second doses are each administered as a single, once-daily (QD) dose.

[0065] In some embodiments, the second doses are each administered as a single, once-weekly (QW) dose.

[0066] In some embodiments, the second doses are each administered as a single, once-daily (QD) oral dose or as a single, once-weekly (QW) oral dose.

[0067] In some embodiments, the second doses are each administered as a single, once-daily (QD) oral dose.

[0068] In some embodiments, the second doses are each administered as a single, once-weekly (QW) oral dose.

[0069] In some embodiments, the second dose is about 5.0 mg / day or less, such as about 5.0 mg / day, about 4.0 mg / day, about 3.0 mg / day, about 2.5 mg / day, about 2.0 mg / day, about 1.75 mg / day, about 1.5 mg / day, about 1.25 mg / day, or about 1.0 mg / day.

[0070] In some embodiments, the second dose is from about 2.5 mg / day to about 7.5 mg / day.

[0071] In some embodiments, the second dose is from about 3.0 mg / day to about 7.0 mg / day.

[0072] In some embodiments, the second dose is about 4.0 mg / day to about 6.0 mg / day.

[0073] In some embodiments, the second dose is about 5.0 mg / day.

[0074] In some embodiments, the second dose is about 4.0 mg / day to about 6.0 mg / day and is administered as a single, once daily (QD) dose.

[0075] In some embodiments, the second dose is about 5.0 mg / day and is administered as a single, once-daily (QD) dose.

[0076] In some embodiments, the second dose is about 4.0 mg / day to about 6.0 mg / day and is administered as a single, once daily (QD) oral dose.

[0077] In some embodiments, the second dose is about 5.0 mg / day and is administered as a single, once daily (QD) oral dose.

[0078] In some embodiments, the second dose is reduced from about 5.0 mg / day or less during the second period.

[0079] In some embodiments, the second dose is reduced during the second period from about 5.0 mg / day or less to about 2.5 mg / day or less, 2.0 mg / day or less, about 1.75 mg / day or less, about 1.5 mg / day or less, about 1.25 mg / day or less, or about 1.0 mg / day or less.

[0080] In some embodiments, the second dose is reduced from about 5.0 mg / day to about 2.5 mg / day, 2.0 mg / day, about 1.75 mg / day, about 1.5 mg / day, about 1.25 mg / day, or about 1.0 mg / day during the second period.

[0081] In some embodiments, the second dose is reduced from about 5.0 mg / day to about 0.75 mg / day to about 1.25 mg / day during the second period.

[0082] In some embodiments, the second dose is reduced from about 5.0 mg / day to about 1.25 mg / day during the second period.

[0083] In some embodiments, the second dose is reduced from about 5.0 mg / day to about 2.0 mg / day, about 1.5 mg / day, about 1.25 mg / day, or about 1.0 mg / day.

[0084] In some embodiments, the second dose is reduced from about 5.0 mg / day to about 1.0 mg / day.

[0085] In some embodiments, the second dose is about 2.5 mg / day or less, such as about 2.5 mg / day, about 2.0 mg / day, about 1.75 mg / day, about 1.5 mg / day, about 1.25 mg / day, or about 1.0 mg / day.

[0086] In some embodiments, the second dose is about 1.0 mg / day to about 2.5 mg / day, for example, about 1.0 mg / day to about 2.0 mg / day, about 1.0 mg / day to about 1.5 mg / day, or about 1.0 mg / day to about 1.25 mg / day.

[0087] In some embodiments, the second dose is from about 2 mg / day to about 2.5 mg / day.

[0088] In some embodiments, the second dose is about 2.5 mg / day.

[0089] In some embodiments, the second dose is about 2 mg / day to about 2.5 mg / day and is administered as a single, once daily (QD) dose.

[0090] In some embodiments, the second dose is about 2.5 mg / day and is administered as a single, once-daily (QD) dose.

[0091] In some embodiments, the second dose is about 2 mg / day to about 2.5 mg / day and is administered as a single, once daily (QD) oral dose.

[0092] In some embodiments, the second dose is about 2.5 mg / day and is administered as a single, once daily (QD) oral dose.

[0093] In some embodiments, the second dose is reduced from about 2.5 mg / day or less during the second period.

[0094] In some embodiments, the second dose is reduced from about 2.5 mg / day or less to about 2.0 mg / day or less, about 1.75 mg / day or less, about 1.5 mg / day or less, about 1.25 mg / day or less, or about 1.0 mg / day or less during the second period.

[0095] In some embodiments, the second dose is reduced during the second period from about 2.5 mg / day to about 2.0 mg / day, about 1.75 mg / day, about 1.5 mg / day, about 1.25 mg / day, or about 1.0 mg / day.

[0096] In some embodiments, the second dose is reduced from about 2.5 mg / day to about 1.25 mg / day during the second period.

[0097] In some embodiments, the second dose is reduced from about 2.5 mg / day to about 2.0 mg / day, about 1.5 mg / day, about 1.25 mg / day, or about 1.0 mg / day.

[0098] In some embodiments, the second dose is reduced from about 2.5 mg / day to about 1.0 mg / day.

[0099] In some embodiments, the reduced dose of the second dose is administered as a single, once-daily (QD) dose.

[0100] In some embodiments, the reduced dose of the second dose is administered as a single, once-daily (QD) oral dose.

[0101] In some embodiments, the second dose is about 50 mg / week or less, e.g., about 40 mg / week or less, about 30 mg / week or less, about 20 mg / week or less, about 10 mg / week or less, about 5 mg / week or less, about 2 mg / week or less, or about 1 mg / week or less.

[0102] In some embodiments, the second dose is about 5 mg / week to about 50 mg / week, for example, about 5 mg / week to about 40 mg / week, about 5 mg / week to about 30 mg / week, about 5 mg / week to about 20 mg / week, about 5 mg / week to about 10 mg / week, about 10 mg / week to about 40 mg / week, about 10 mg / week to about 30 mg / week, about 10 mg / week to about 20 mg / week, about 20 mg / week to about 40 mg / week, about 20 mg / week to about 30 mg / week, or about 30 mg / week to about 40 mg / week.

[0103] In some embodiments, the second dose is about 18 mg / week to about 20 mg / week.

[0104] In some embodiments, the second dose is about 20 mg / week.

[0105] In some embodiments, the second dose is about 18 mg / week to about 20 mg / week and is administered as a single, once-weekly (QW) dose.

[0106] In some embodiments, the second dose is about 20 mg / week and is administered as a single, once-weekly (QW) dose.

[0107] In some embodiments, the second dose is reduced from about 50 mg / week or less during the second period.

[0108] In some embodiments, the second dose is reduced from about 50 mg / week or less to about 40 mg / week or less, about 30 mg / week or less, about 20 mg / week or less, about 10 mg / week or less, about 5 mg / week or less, about 2 mg / week or less, or about 1 mg / week or less during the second period.

[0109] In some embodiments, the second dose is decreased during the second period from about 5 mg / week to about 50 mg / week to about 5 mg / week to about 40 mg / week, about 5 mg / week to about 30 mg / week, about 5 mg / week to about 20 mg / week, about 5 mg / week to about 10 mg / week, about 10 mg / week to about 40 mg / week, about 10 mg / week to about 30 mg / week, about 10 mg / week to about 20 mg / week, about 20 mg / week to about 40 mg / week, about 20 mg / week to about 30 mg / week, or about 30 mg / week to about 40 mg / week.

[0110] In some embodiments, the second dose is reduced from about 50 mg / week to about 40 mg / week, about 30 mg / week, about 20 mg / week, about 10 mg / week, or about 5 mg / week during the second period.

[0111] In some embodiments, the second dose is reduced from about 20 mg / week to about 10 mg / week during the second period.

[0112] In some embodiments, the second dose is reduced from about 20 mg / week to about 10 mg / week during the second period.

[0113] In some embodiments, the second dose is reduced from about 10 mg / week to about 5 mg / week during the second period.

[0114] In some embodiments, the reduced dose of the second dose is administered as a single, once-weekly (QW) dose.

[0115] In some embodiments, the patient has been identified as showing one or more symptoms associated with one or more adverse events (TEAEs) occurring during treatment. As used herein, the severity of TEAEs described herein is assessed using CommonTerminology Criteria for Adverse Events 4.03 version, the disclosure of which is incorporated herein by reference in its entirety.

[0116] In some embodiments, the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs) during the first period.

[0117] In some embodiments, when the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs), the first dose is reduced from about 3 mg / day to about 50 mg / day to about 3 mg / day to about 30 mg / day, about 3 mg / day to about 20 mg / day, about 3 mg / day to about 15 mg / day, about 3 mg / day to about 10 mg / day, about 3 mg / day to about 7 mg / day, or about 3 mg / day to about 5 mg / day.

[0118] In some embodiments, the first dose is reduced from about 50 mg / day to about 30 mg / day, about 20 mg / day, about 15 mg / day, about 10 mg / day, about 5 mg / day, about 2 mg / day, or about 1 mg / day during the first period when the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs).

[0119] In some embodiments, the first dose is reduced from about 20 mg / day to about 15 mg / day, about 10 mg / day, about 5 mg / day, about 2 mg / day, or about 1 mg / day during the first period when the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs).

[0120] In some embodiments, the first dose is reduced from about 20 mg / day to about 10 mg / day when the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs).

[0121] In some embodiments, the first dose is reduced from about 20 mg / day to about 5 mg / day when the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs).

[0122] In some embodiments, the first dose is reduced from about 10 mg / day to about 5 mg / day when the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs).

[0123] In some embodiments, the methods provided herein further comprise discontinuing administration of the first dose when the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs).

[0124] In some embodiments, the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs) during the second period.

[0125] In some embodiments, when the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs), the second dose is reduced from about 3 mg / day to about 50 mg / day to about 3 mg / day to about 30 mg / day, about 3 mg / day to about 20 mg / day, about 3 mg / day to about 15 mg / day, about 3 mg / day to about 10 mg / day, about 3 mg / day to about 7 mg / day, or about 3 mg / day to about 5 mg / day during the second period.

[0126] In some embodiments, when the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs), the second dose is reduced from about 50 mg / week to about 40 mg / week, about 30 mg / week, about 20 mg / week, about 10 mg / week, about 5 mg / week, about 2 mg / week, or about 1 mg / week during the second period.

[0127] In some embodiments, when the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs), the second dose is reduced during the second period from about 50 mg / week or less to about 40 mg / week or less, about 30 mg / week or less, about 20 mg / week or less, about 10 mg / week or less, about 5 mg / week or less, about 2 mg / week or less, or about 1 mg / week or less.

[0128] In some embodiments, when the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs), the second dose is reduced from about 20 mg / week to about 10 mg / week or less during the second period.

[0129] In some embodiments, when the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs), the second dose is reduced from about 20 mg / week to about 5 mg / week or less during the second period.

[0130] In some embodiments, when the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs), the second dose is reduced from about 10 mg / week to about 5 mg / week or less during the second period.

[0131] In some embodiments, one or more treatment-emergent adverse events include one or more of the following: diarrhea / colitis, nausea, fatigue, rash, neutropenia, pyrexia, hypotension, sepsis, respiratory failure, pneumonitis, pneumonia, hypertension, hyperglycemia, abdominal pain, bronchitis, dehydration, thrombocytopenia, cough, vomiting, anorexia, increased lacrimation, oral herpes, tachycardia, spinal cord compression, intractable pain, increased alkaline phosphatase, increased transaminases, hyperlipidemia, hypercalcemia, dizziness, alopecia, constipation, fluid overload, headache, hypokalemia, night sweats, encephalopathy, atrial flutter, atrial fibrillation, and dyspnea.

[0132] In some embodiments, symptoms associated with one or more treatment-emergent adverse events are symptoms associated with one or more of the following: diarrhea / colitis, nausea, fatigue, rash, neutropenia, pyrexia, hypotension, sepsis, respiratory failure, parenchymal lung inflammation, pneumonia, hypertension, hyperglycemia, abdominal pain, bronchitis, dehydration, thrombocytopenia, cough, vomiting, anorexia, increased lacrimation, oral herpes, tachycardia, spinal cord compression, intractable pain, increased alkaline phosphatase, increased transaminases, hyperlipidemia, hypercalcemia, vertigo, alopecia, constipation, fluid overload, headache, hypokalemia, night sweats, encephalopathy, atrial flutter, atrial fibrillation, and dyspnea.

[0133] In some embodiments, one or more treatment-emergent adverse events include one or more of the following: diarrhea / colitis, nausea, fatigue, rash, cough, vomiting, dizziness, fever, hypokalemia, abdominal pain, constipation, decreased appetite, night sweats, pruritus, back pain, chills, leukopenia, neutropenia, lymphopenia, thrombocytopenia, and anemia.

[0134] In some embodiments, one or more treatment-emergent adverse events include one or more of diarrhea / colitis and rash.

[0135] In some embodiments, diarrhea / colitis includes one or more of diarrhea, colitis, enterocolitis, gastroenteritis, microscopic colitis, and cytomegalovirus colitis.

[0136] In some embodiments, the rash comprises one or more of: exfoliative dermatitis, rash, erythematous rash, macular rash, maculopapular rash, pruritic rash, exfoliative rash, generalized rash, epidemic rash, and pustular rash.

[0137] In some embodiments, neutropenia comprises febrile neutropenia.In some embodiments, elevated transaminases comprise elevated alanine aminotransferase (ALT), elevated aspartate aminotransferase (AST), or a combination thereof.

[0138] As used herein, the term "contacting" refers to bringing the indicated moieties together in an in vitro system or an in vivo system. For example, "contacting" of PI3K with a PI3Kδ inhibitor compound described herein includes administering a compound described herein to an individual or patient such as a human having PI3K, and, for example, introducing a compound described herein into a sample containing cells or purified preparations containing PI3K.

[0139] In some embodiments, the PI3Kδ inhibitor is a compound of U.S. Patent Publication No. US2011 / 0015212, filed June 28, 2010, U.S. Patent Publication No. 2013 / 0059835, filed August 31, 2013, U.S. Patent Publication No. 2011 / 0183985, filed December 17, 2010, U.S. Patent Publication No. 2012 / 0157430, filed December 19, 2011, U.S. Patent Publication No. 2014 / 0249132, filed February 28, 2014, or U.S. Patent Publication No. 2016 / 0257689, filed February 26, 2016, each of which is herein incorporated by reference in its entirety.

[0140] In some embodiments, the PI3Kδ inhibitor is prepared by the method of U.S. Patent Publication No. US2011 / 0015212 filed on June 28, 2010, U.S. Patent Publication No. 2013 / 0059835 filed on August 31, 2013, U.S. Patent Publication No. 2011 / 0183985 filed on December 17, 2010, or U.S. Patent Publication No. 2012 / 0157430 filed on December 19, 2011, U.S. Patent Publication No. 2014 / 0249132 filed on February 28, 2014, or U.S. Patent Publication No. 2016 / 0257689 filed on February 26, 2016, each of which is herein incorporated by reference in its entirety.

[0141] In some embodiments, the PI3Kδ inhibitor is selected from:

[0142] 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one; and

[0143] 5-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}-1,3-oxazolidin-2-one;

[0144] or a pharmaceutically acceptable salt thereof.

[0145] In some embodiments, the PI3Kδ inhibitor is 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one or a pharmaceutically acceptable salt thereof.

[0146] In some embodiments, the PI3Kδ inhibitor is selected from:

[0147] (S)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one;

[0148] (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one;

[0149] (S)-4-(3-((R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; and

[0150] (R)-4-(3-((R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one;

[0151] or a pharmaceutically acceptable salt thereof.

[0152] In some embodiments, the PI3Kδ inhibitor is a pharmaceutically acceptable salt of 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one.

[0153] In some embodiments, the PI3Kδ inhibitor is selected from a pharmaceutically acceptable salt of:

[0154] (S)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one;

[0155] (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one;

[0156] (S)-4-(3-((R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; and

[0157] (R)-4-(3-((R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one.

[0158] In some embodiments, the PI3Kδ inhibitor is a pharmaceutically acceptable salt of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one.

[0159] In some embodiments, the PI3Kδ inhibitor is (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride. In some embodiments, the salt is a salt in which the stoichiometric ratio of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one to hydrochloric acid is 1:1.

[0160] In some embodiments, the salt is crystalline (for methods and processes for preparing (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride, see, e.g., U.S. Patent Publication No. 2016 / 0257689 filed on February 26, 2016. U.S. Patent Publication No. 2016 / 0257689 is incorporated herein by reference in its entirety.).

[0161] In some embodiments, the hydrochloride salt of 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one has: at least one, two, three, four or five XRPD peaks, expressed in 2-θ, selected from about 11.3°, about 16.4°, about 21.0°, about 23.0°, about 28.1°, about 29.8°, about 30.7°, about 31.6°, about 32.9°, about 34.7°, about 36.9°, about 38.8°, about 39.9°, about 40.7°, about 41.9°, about 42.9°, about 43.7°, about 44.9°, about 45.7°, at least two XRPD peaks, expressed as 2-θ, selected from about 11.3°, about 16.4°, about 21.0°, about 23.0°, about 28.1°, about 31.2°, and about 32.8°; at least three XRPD peaks, expressed as 2-θ, selected from about 11.3°, about 16.4°, about 21.0°, about 23.0°, about 28.1°, about 31.2°, and about 32.8°. In some embodiments, the hydrochloride salt of the compound of Formula I has at least four XRPD peaks, expressed as 2-θ, selected from about 11.3°, about 16.4°, about 21.0°, about 23.0°, about 28.1°, about 31.2°, and about 32.8°. In some embodiments, the hydrochloride salt of 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one has all of the listed XRPD peaks, expressed as 2-theta, at about 11.3°, about 16.4°, about 21.0°, about 23.0°, about 28.1°, about 31.2°, and about 32.8°. In some embodiments, the hydrochloride salt of 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one has a DSC thermogram with an endothermic peak at about 207°C.

[0162] In some embodiments, salts and compounds described herein are substantially separated. "Substantially separated" refers to that a salt or compound is at least partially or substantially separated from the environment in which it is formed or detected. Partial separation can include, for example, a composition enriched in the form of a salt as described herein. Substantially separating can include a composition containing at least about 50% by weight, at least about 60% by weight, at least about 70% by weight, at least about 80% by weight, at least about 90% by weight, at least about 95% by weight, at least about 97% by weight, or at least about 99% by weight of a salt as described herein or a salt thereof. Methods for separating compounds and their salts are conventional in the art.

[0163] In some embodiments, the PI3Kδ inhibitor is 5-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}-1,3-oxazolidin-2-one or a pharmaceutically acceptable salt thereof.

[0164] In some embodiments, the PI3Kδ inhibitor is selected from:

[0165] (R)-5-{3-[(R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}-1,3-oxazolidin-2-one;

[0166] (R)-5-{3-[(S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}-1,3-oxazolidin-2-one;

[0167] (S)-5-{3-[(S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}-1,3-oxazolidin-2-one; and

[0168] (S)-5-{3-[(R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}-1,3-oxazolidin-2-one;

[0169] or a pharmaceutically acceptable salt thereof.

[0170] The compounds described herein (including salts thereof) can be prepared using known organic synthesis techniques and can be synthesized according to any of a number of possible synthetic routes. In some embodiments, the compounds can be prepared as described in U.S. Patents 9,199,982 and 9,932,341, each of which is incorporated herein by reference in its entirety.

[0171] The reaction for preparing the compounds described herein can be carried out in a suitable solvent, which can be easily selected by a technician in the field of organic synthesis. Suitable solvents can be substantially free from reacting with starting materials (reactants), intermediates or products at the temperature of the reaction (e.g., a temperature that can range from the freezing temperature of the solvent to the boiling temperature of the solvent). A given reaction can be carried out in a solvent or a mixture of more than one solvent. Depending on the specific reaction steps, suitable solvents for specific reaction steps can be selected by a skilled technician.

[0172] The preparation of the compounds described herein may involve the protection and deprotection of various chemical groups. The need for protection and deprotection and the selection of appropriate protecting groups can be easily determined by those skilled in the art. The chemical properties of protecting groups can be found in, for example, TW Greene and PGM Wuts, Protective Groups in Organic Synthesis, 3rd Edition, Wiley & Sons, Inc., New York (1999), which is incorporated herein by reference in its entirety.

[0173] The reaction can be monitored by any suitable method known in the art. For example, product formation can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., 1 H or 13 C), infrared spectroscopy, spectrophotometry (e.g., UV-visible light), mass spectrometry; or chromatographic methods, such as high performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LCMS) or thin layer chromatography (TLC). The compound can be purified by a variety of methods including high performance liquid chromatography (HPLC) ("Preparative LC-MS Purification: Improved Compound Specific Method Optimization" Karl F. Blom, Brian Glass, Richard Sparks, Andrew P. Combs J. Combi. Chem. 2004, 6 (6), 874-883, which is incorporated herein by reference in its entirety) and normal phase silica gel chromatography.

[0174] The compounds described herein can regulate the activity of one or more of various kinases, such as phosphoinositide 3-kinase (PI3K). The term "regulation" refers to the ability to increase or decrease the activity of one or more members of the PI3K family. Therefore, the compounds described herein can be used in the method of regulating PI3K, and the regulation is achieved by contacting PI3K with any one or more of the compounds or compositions described herein. In some embodiments, the compounds of the present invention can serve as inhibitors of one or more PI3K. In other embodiments, the compounds described herein can be used to regulate the activity of PI3K in individuals who need to regulate receptors by administering a regulated amount of the compounds described herein or a pharmaceutically acceptable salt thereof. In some embodiments, regulation is inhibition. In view of the fact that the growth and survival of cancer cells are affected by multiple signal transduction pathways, the present invention is suitable for treating disease states characterized by drug-resistant kinase mutations. In addition, different kinase inhibitors can be used in combination, and they show different preferences in kinases whose activity is regulated by the inhibitor. This method can show high efficiency in treating disease states by targeting multiple signal transduction pathways, reducing the possibility of drug resistance in cells and reducing the toxicity used for the treatment of diseases.

[0175] The PI3Kδ inhibitors described herein may be selective. "Selectivity" refers to that the compound binds or inhibits a kinase with greater affinity or potency, respectively, compared to at least one other kinase. In some embodiments, the compounds described herein are selective inhibitors of PI3Kδ relative to PI3Kγ, PI3Kα and / or PI3Kβ. In some embodiments, the compounds described herein are selective inhibitors of PI3Kδ (e.g., relative to PI3Kα, PI3Kβ and PI3Kγ). In some embodiments, the selectivity may be at least about 2 times, 5 times, 10 times, at least about 20 times, at least about 50 times, at least about 100 times, at least about 200 times, at least about 500 times, or at least about 1000 times. Selectivity may be measured by conventional methods in the art. In some embodiments, selectivity may be measured at the K of each enzyme. m In some embodiments, the selectivity of the compounds described herein can be determined by a cellular assay associated with a specific PI3K kinase activity.

[0176] The kinases that the compounds of the present invention bind to and / or regulate (e.g., inhibit) include any member of the PI3K family. In some embodiments, PI3K is PI3Kα, PI3Kβ, PI3Kγ, or PI3Kδ. In some embodiments, PI3K is PI3Kγ or PI3Kδ. In some embodiments, PI3K is PI3Kγ. In some embodiments, PI3K is PI3Kδ. In some embodiments, PI3K includes mutations. Mutations can be substitutions of one amino acid for another amino acid, or the absence of one or more amino acids. In such embodiments, mutations can be present in the kinase domain of PI3K.

[0177] In some embodiments of the methods of the invention, more than one compound described herein is used to inhibit the activity of one kinase (eg, PI3Kγ or PI3Kδ).

[0178] In some embodiments of the methods of the invention, more than one compound described herein is used to inhibit more than one kinase, such as at least two kinases (eg, PI3Kγ and PI3Kδ).

[0179] In some embodiments of the methods of the invention, one or more compounds are used in combination with another kinase inhibitor to inhibit the activity of one kinase (eg, PI3Kγ or PI3Kδ).

[0180] In some embodiments of the methods of the invention, one or more compounds are used in combination with another kinase inhibitor to inhibit the activity of more than one kinase (eg, PI3Kγ or PI3Kδ), such as the activity of at least two kinases.

[0181] Another aspect of the present invention relates to a method for treating a kinase (such as PI3Kδ)-related disease or condition in an individual (e.g., a patient) by administering a therapeutically effective amount or dose of one or more compounds of the present invention or a pharmaceutical composition thereof to an individual in need of such treatment. PI3K-related diseases may include any disease, disorder, or condition directly or indirectly associated with the expression or activity of PI3K, including overexpression and / or abnormal activity levels. In some embodiments, the disease may be associated with Akt (protein kinase B), mammalian target of rapamycin (mTOR), or phosphoinositide-dependent kinase 1 (PDK1). In some embodiments, mTOR-related diseases may be inflammation, atherosclerosis, psoriasis, restenosis, benign prostatic hypertrophy, bone disorders, pancreatitis, angiogenesis, diabetic retinopathy, atherosclerosis, arthritis, immune disorders, kidney disease, or cancer. PI3K-related diseases may also include any disease, disorder, or condition that can be prevented, improved, or cured by regulating PI3K activity. In some embodiments, the disease is characterized by abnormal activity of PI3K. In some embodiments, the disease is characterized by mutant PI3K. In such embodiments, the mutation may be present in the kinase domain of PI3K.

[0182] Examples of PI3K-associated diseases include immune-based diseases involving the system including, for example, rheumatoid arthritis, allergies, asthma, glomerulonephritis, lupus, or inflammation associated with any of the foregoing.

[0183] Other examples of PI3K-associated diseases include cancers such as breast cancer, prostate cancer, colon cancer, endometrial cancer, brain cancer, bladder cancer, skin cancer, uterine cancer, ovarian cancer, lung cancer, pancreatic cancer, kidney cancer, stomach cancer, or blood cancer.

[0184] In some embodiments, the blood cancer is acute myeloblastic leukemia (AML) or chronic myeloid leukemia (CML), or B-cell lymphoma. In some embodiments, the B-cell lymphoma is diffuse large B-cell lymphoma.

[0185] Other examples of PI3K-associated diseases include lung diseases, such as acute lung injury (ALI) and adult respiratory distress syndrome (ARDS).

[0186] Other examples of PI3K-related diseases include osteoarthritis, restenosis, atherosclerosis, bone disorders, arthritis, diabetic retinopathy, psoriasis, benign prostatic hypertrophy, inflammation, angiogenesis, pancreatitis, kidney disease, inflammatory bowel disease, myasthenia gravis, multiple sclerosis or Sjögren's syndrome, etc.

[0187] Other examples of PI3K-associated diseases include autoimmune hemolytic anemia, polycythemia vera, and pemphigus.

[0188] In some embodiments, the disease is non-Hodgkin lymphoma (NHL). In some embodiments, non-Hodgkin lymphoma (NHL) is relapsed or refractory NHL or relapsed follicular NHL.

[0189] In some embodiments, the disease is an aggressive lymphoma (eg, germinal center B-cell-like (GCB) or activated B-cell-like (ABC)).

[0190] In some embodiments, the disease is selected from diffuse large B-cell lymphoma, chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma, Hodgkin's lymphoma, Burkitt's lymphoma, Waldenstrom's macroglobulinemia, macroglobulinemia), prolymphocytic leukemia, acute lymphoblastic leukemia, myelofibrosis, mucosa-associated lymphoid tissue (MALT) lymphoma, mediastinal (thymic) large B-cell lymphoma, lymphomatoid granulomatosis, splenic marginal zone lymphoma, primary effusion lymphoma, intravascular large B-cell lymphoma, plasma cell leukemia, extramedullary plasmacytoma, smoldering myeloma (also called asymptomatic myeloma), monoclonal gammopathy of undetermined significance (MGUS), activated B-cell-like (ABC) diffuse large B-cell lymphoma (ABC-DLBCL), and germinal center B-cell (GCB) diffuse large B-cell lymphoma (GCB-DLBCL).

[0191] In some embodiments, the disease is myelofibrosis. In some embodiments, myelofibrosis is selected from post-essential thrombocythaemia myelofibrosis (PET-MF), primary myelofibrosis (PMF) and post-polycythaemia vera myelofibrosis (PPV-MF). In some embodiments, myelofibrosis is post-essential thrombocythaemia myelofibrosis (PET-MF). In some embodiments, myelofibrosis is primary myelofibrosis (PMF). In some embodiments, myelofibrosis is post-polycythaemia vera myelofibrosis (PPV-MF).

[0192] In some embodiments, the disease is acute myeloid leukemia. In some embodiments, the disease is Burkitt's lymphoma. In some embodiments, the disease is activated B-cell-like (ABC) diffuse large B-cell lymphoma (ABC-DLBCL) and germinal center B-cell (GCB) diffuse large B-cell lymphoma (GCB-DLBCL).

[0193] In some embodiments, the disease is selected from chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Hodgkin lymphoma (HL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), and Waldenstrom's macroglobulinemia (WM).

[0194] In some embodiments, the marginal zone lymphoma (MZL) is selected from extranodal MZL, nodal MZL, splenic MZL, and unknown MZL subtype.

[0195] In some embodiments, the Hodgkin lymphoma (HL) is selected from classical Hodgkin lymphoma (HL) and nodular lymphocyte-predominant HL.

[0196] In some embodiments, the diffuse large B-cell lymphoma is selected from activated B-cell-like (ABC) diffuse large B-cell lymphoma (ABC-DLBCL) and germinal center B-cell (GCB) diffuse large B-cell lymphoma (GCB-DLBCL).

[0197] In some embodiments, the disease is selected from autoimmune hemolytic anemia, polycythemia vera, and pemphigus.

[0198] In some embodiments, the present application also provides a method for treating a disease in a patient, wherein the disease is selected from chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Hodgkin lymphoma (HL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL) and Waldenstrom's macroglobulinemia (WM), the method comprising:

[0199] i) administering to the patient a first dose of about 20 mg / day to about 50 mg / day of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride over a first period of about 8 weeks to about 9 weeks; and

[0200] ii) administering to the patient a second dose of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride in a second period occurring after the first period, wherein the second dose is about 2.5 mg / day or less.

[0201] In some embodiments, the present application also provides a method for treating a disease in a patient, wherein the disease is selected from chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Hodgkin lymphoma (HL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL) and Waldenstrom's macroglobulinemia (WM), the method comprising:

[0202] i) administering to the patient a first dose of about 20 mg / day to about 50 mg / day of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride over a first period of about 8 weeks to about 9 weeks; and

[0203] ii) administering to the patient a second dose of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride in a second period occurring after the first period, wherein the second dose is from about 20 mg / week to about 50 mg / week.

[0204] In some embodiments, the present application also provides a method for treating a disease in a patient, wherein the disease is selected from chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Hodgkin lymphoma (HL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL) and Waldenstrom's macroglobulinemia (WM), the method comprising:

[0205] i) administering to the patient a first dose of about 20 mg / day of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride over a first period of about 8 weeks; and

[0206] ii) administering to the patient a second dose of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride in a second period occurring after the first period, wherein the second dose is about 2.5 mg / day.

[0207] In some embodiments, the present application also provides a method for treating a disease in a patient, wherein the disease is selected from chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Hodgkin lymphoma (HL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL) and Waldenstrom's macroglobulinemia (WM), the method comprising:

[0208] i) administering to the patient a first dose of about 20 mg / day of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride over a first period of about 8 weeks; and

[0209] ii) administering to the patient a second dose of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride in a second period occurring after the first period, wherein the second dose is about 20 mg / week.

[0210] As used herein, the terms "subject" or "patient" are used interchangeably and refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, and most preferably humans.

[0211] As used herein, the phrase "therapeutically effective amount" refers to that amount of an active compound or agent that elicits the biological or medical response in a tissue, system, animal, individual or human that is being sought by the researcher, veterinarian, medical doctor or other clinician.

[0212] As used herein, the terms "treating" or "treatment" refer to one or more of the following: (1) preventing a disease; e.g., preventing a disease, condition, or disorder in an individual who may be susceptible to the disease, condition, or disorder but is not yet experiencing or showing the pathology or symptomology of the disease; (2) inhibiting a disease; e.g., inhibiting a disease, condition, or disorder in an individual who is experiencing or showing the pathology or symptomology of the disease, condition, or disorder (i.e., curbing further development of the pathology and / or symptomology); and (3) ameliorating a disease; e.g., ameliorating a disease, condition, or disorder in an individual who is experiencing or showing the pathology or symptomology of the disease, condition, or disorder (i.e., reversing the pathology and / or symptomology), such as reducing the severity of the disease. In some embodiments, the terms "treating" or "treatment" refer to inhibiting or ameliorating a disease.

[0213] As used herein, "QD" is intended to mean a dose administered to a subject once daily. "QW" is intended to mean a dose administered to a subject once weekly. "Q2W" is intended to mean a dose administered to a subject once every other week. "Q3W" is intended to mean a dose administered to a subject once every three weeks. "Q4W" is intended to mean a dose administered to a subject once every four weeks.

[0214] As used herein, "about" is intended to encompass a deviation of ±10% when referring to a measurable value such as an amount, dosage, duration, etc. In certain embodiments, "about" may include deviations of ±5%, ±1%, or ±0.1% from the stated value, and any deviations therebetween, as such deviations are appropriate for performing the disclosed methods.

[0215] Combination therapy

[0216] I. Immune Checkpoint Therapy

[0217] In some embodiments, the PI3K inhibitors provided herein can be used in combination with one or more immune checkpoint inhibitors to treat cancer as described herein. In one embodiment, the combination with one or more immune checkpoint inhibitors as described herein can be used to treat B cell malignancies as described herein. The compounds disclosed herein can be used in combination with one or more immune checkpoint inhibitors. Exemplary immune checkpoint inhibitors include inhibitors for immune checkpoint molecules such as: CD20, CD28, CD40, CD122, CD96, CD73, CD47, GITR, CSF1R, JAK, PI3Kδ, PI3Kγ, TAM, arginase, HPK1, CD137 (also known as 4-1BB), ICOS, B7-H3, B7-H4, BTLA, CTLA-4, LAG3, TIM3, VISTA, TIGIT, PD-1, PD-L1 and PD-L2. In some embodiments, the immune checkpoint molecule is a stimulatory checkpoint molecule selected from the following: CD27, CD28, CD40, ICOS, OX40, GITR and CD137. In some embodiments, the immune checkpoint molecule is an inhibitory checkpoint molecule selected from the following: A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, TIM3, TIGIT and VISTA. In some embodiments, the compounds of the present disclosure provided herein can be used in combination with one or more agents selected from the following: KIR inhibitors, TIGIT inhibitors, LAIR1 inhibitors, CD160 inhibitors, 2B4 inhibitors and TGFRβ inhibitors.

[0218] In some embodiments, the PI3K inhibitors provided herein can be used in combination with agonists of one or more of the following immune checkpoint molecules: for example, OX40, CD27, OX40, GITR, and CD137 (also known as 4-1BB).

[0219] In some embodiments, the inhibitor of an immune checkpoint molecule is an anti-PD1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody.

[0220] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of PD-1, such as an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab, pembrolizumab (also known as MK-3475), durvalumab In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab or pembrolizumab. In some embodiments, the anti-PD1 antibody is pembrolizumab. In some embodiments, the anti-PD-1 monoclonal antibody is MGA012. In some embodiments, the anti-PD1 antibody is SHR-1210. Other anticancer agents include antibody therapeutics such as 4-1BB (e.g., urelumab, utomilumab).

[0221] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of PD-L1, such as an anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-L1 monoclonal antibody is BMS-935559, MEDI4736, MPDL3280A (also known as RG7446), or MSB0010718C. In some embodiments, the anti-PD-L1 monoclonal antibody is MPDL3280A or MEDI4736.

[0222] In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of PD-1 and PD-L1, such as an anti-PD-1 / PD-L1 monoclonal antibody. In some embodiments, the anti-PD-1 / PD-L1 is MCLA-136.

[0223] In some embodiments, the inhibitor is MCLA-145.

[0224] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of CTLA-4, such as an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab, tremelimumab, AGEN1884 or CP-675,206.

[0225] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of LAG3, such as an anti-LAG3 antibody. In some embodiments, the anti-LAG3 antibody is BMS-986016, LAG525 or INCAGN2385.

[0226] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of TIM3, such as an anti-TIM3 antibody. In some embodiments, the anti-TIM3 antibody is INCAGN2390, MBG453 or TSR-022.

[0227] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of GITR, such as an anti-GITR antibody. In some embodiments, the anti-GITR antibody is TRX518, MK-4166, INCAGN1876, MK-1248, AMG228, BMS-986156, GWN323 or MEDI1873.

[0228] In some embodiments, the inhibitor of the immune checkpoint molecule is an agonist of OX40, such as an OX40 agonist antibody or an OX40L fusion protein. In some embodiments, the anti-OX40 antibody is MEDI0562, MOXR-0916, PF-04518600, GSK3174998, or BMS-986178. In some embodiments, the OX40L fusion protein is MEDI6383.

[0229] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of CD20, such as an anti-CD20 antibody. In some embodiments, the anti-CD20 antibody is obinutuzumab or rituximab.

[0230] The compounds of the present disclosure can be used in combination with bispecific antibodies. In some embodiments, one domain of the bispecific antibody targets PD-1, PD-L1, CTLA-4, GITR, OX40, TIM3, LAG3, CD137, ICOS, CD3 or TGFβ receptor.

[0231] In some embodiments, the compounds of the present disclosure can be used in combination with one or more metabolic enzyme inhibitors. In some embodiments, the metabolic enzyme inhibitor is an inhibitor of IDO1, TDO or arginase. Examples of IDO1 inhibitors include epacadostat, NLG919, BMS-986205, PF-06840003, IOM2983, RG-70099 and LY338196.

[0232] As provided throughout, the additional compounds, inhibitors, agents, etc. can be combined with the compounds of the present invention in a single or sequential dosage form, or they can be administered simultaneously or sequentially as separate dosage forms.

[0233] II. Cancer Therapy

[0234] The growth and survival of cancer cells can be affected by multiple signaling pathways. Therefore, it is useful to combine different enzyme / protein / receptor inhibitors that exhibit different preferences for targets whose activity they modulate in order to treat such conditions. Targeting more than one signaling pathway (or more than one biomolecule involved in a given signaling pathway) can reduce the likelihood of resistance developing in a cell population and / or reduce the toxicity of the treatment.

[0235] The compounds of the present disclosure may be used in combination with one or more other enzyme / protein / receptor inhibitors or one or more therapies for treating a disease such as cancer. Examples of diseases and indications that may be treated with combination therapy include those described herein.

[0236] One or more additional agents or therapeutic agents such as chemotherapeutics, immuno-oncology agents, metabolic enzyme inhibitors, chemokine receptor inhibitors and phosphatase inhibitors and targeted therapies such as Bcr-Abl, Flt-3, EGFR, HER2, JAK, c-MET, VEGFR, PDGFR, c-Kit, IGF-1R, RAF and FAK kinase inhibitors. One or more additional agents can be administered to the patient simultaneously or sequentially.

[0237] Exemplary antibodies for use in combination therapy include, but are not limited to, Trastuzumab (e.g., anti-HER2), Ranibizumab (e.g., anti-VEGF-A), Bevacizumab (trade name Avastin, e.g., anti-VEGF), Panitumumab (e.g., anti-EGFR), Cetuximab (e.g., anti-EGFR), Rituxan (anti-CD20), and antibodies to c-MET.

[0238] One or more of the following agents may be used in combination with the compounds of the present disclosure and are provided as a non-limiting list: cytostatics, taxotere, paclitaxel, camptostar, epothilones, 5-fluorouracil, SCH66336, R115777, L778,123, BMS214662, IRESSA TM (gefitinib), TARCEVA TM (erlotinib), EGFR antibody, GLEEVEC TM (imatinib mesylate), intron, cytarabine, doxorubicin, cyclophosphamide, nitrogen mustard, triethylenemelamine, triethylenethiophosphoramide, busulfan, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, ELOXATIN TM (oxaliplatin), vindesine, mithramycin, deoxycoformycin, L-asparaginase, 17α-ethinyl estradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone, aminoglutethimide, medroxyprogesterone, leuprolide, flutamide, goserelin, hydroxyurea, amsacrine, navelbene, reloxafine, droloxafine, hexamethonium, acrivastine, HERCEPTIN TM (trastuzumab), BEXXAR TM (tositumomab), VELCADE TM (bortezomib), ZEVALIN TM (ibritumomab tiuxetan), TRISENOX TM (arsenic trioxide), XELODA TM (capecitabine), vinorelbine, porfimer, ERBITUX TM (cetuximab), aphidicolon, rituxan, Sml1, triapine, didox, trimidox, amidox, 3-AP and MDL-101,731.

[0239] The compounds disclosed herein can also be used in combination with other methods for treating cancer: for example, chemotherapy, radiotherapy, tumor-targeted therapy, adjuvant therapy, immunotherapy or surgery. Examples of immunotherapy include cytokine therapy (e.g., interferon, GM-CSF, G-CSF, IL-2), CRS-207 immunotherapy, cancer vaccines, monoclonal antibodies, adoptive T cell transfer, Toll receptor agonists, STING agonists, oncolytic virus therapy and immunomodulatory small molecules, including thalidomide or JAK1 / 2 inhibitors, etc. The compound can be administered in combination with one or more anticancer drugs (such as chemotherapeutic agents). Exemplary chemotherapeutic agents include any of the following: abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab, bexarotene, baricitinib, bleomycin, bortezombi, bortezomib, busulfan intravenous, busulfan oral oral), calusterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparin sodium, dasatinib, daunorubicin, decitabine, denileukin, denileukin-diphtheria toxin conjugatediftitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone propionate, eculizumab, epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate acetate, histrelin acetate, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib mesylate, interferon alfa 2a, irinotecan, lapatinib ditosylate, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, nitrogen mustard, megestrol acetate, melphalan, mercaptopurine, methotrexate, methoxsalen, mitomycin CC), mitotane, mitoxantrone, nandrolone phenpropionate, nelarabine, nofetumomab, olaparib, oxaliplatin, paclitaxel, pamidronate, panitumumab, pegaspargase, pegfilgrastim, pemetrexed disodium, pentostatin, pipobroman, plicamycin, procarbazine, quinacrine, rasburicase, rituximab, ruxolitinib, rucaparib, sorafenib, streptozocin, sunitinib, sunitinib malate maleate, tamoxifen, temozolomide, teniposide, testolactone, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, niraparib, veliparib, talazoparib, and zoledronate.

[0240] Additional examples of chemotherapeutic agents include proteasome inhibitors (eg, bortezomib), thalidomide, revlimid; and DNA damaging agents such as melphalan, doxorubicin, cyclophosphamide, vincristine, etoposide, carmustine, and the like.

[0241] Exemplary Bcr-Abl inhibitors include compounds and pharmaceutically acceptable salts.

[0242] Exemplary suitable Flt-3 inhibitors include compounds and pharmaceutically acceptable salts thereof.

[0243] Exemplary suitable RAF inhibitors include compounds and pharmaceutically acceptable salts thereof.

[0244] Exemplary suitable FAK inhibitors include compounds and pharmaceutically acceptable salts thereof.

[0245] In some embodiments, the compounds of the present disclosure may be used in combination with one or more other kinase inhibitors, including imatinib, particularly for treating patients who are resistant to imatinib or other kinase inhibitors.

[0246] The compound can be used in combination with tumor vaccines and CAR (chimeric antigen receptor) T cell therapy as a T cell activation enhancer. In some embodiments, tumor vaccines include proteins from viruses related to the following human cancers: such as human papillomavirus (HPV), hepatitis viruses (HBV and HCV) and Kaposi's herpes sarcoma virus (KHSV). In some embodiments, the compounds of the present disclosure can be used in combination with tumor-specific antigens such as heat shock proteins separated from tumor tissue itself. In some embodiments, the compound can be combined with dendritic cell immunity to activate effective anti-tumor responses.

[0247] Methods for the safe and effective administration of most of these chemotherapeutic agents are known to those skilled in the art. In addition, their administration is described in standard literature. For example, the administration of a variety of chemotherapeutic agents is described in "Physicians' Desk Reference" (PDR, e.g., 1996 edition, Medical Economics Company, Montvale, NJ), the disclosure of which is incorporated herein by reference, as if set forth in full.

[0248] In some embodiments, the additional therapeutic agent is selected from a JAK inhibitor (eg, a selective JAK1 inhibitor), a pan-PIM selective kinase inhibitor, an inhibitor of an epigenetic regulator, and an immunotherapeutic agent.

[0249] In some embodiments, the inhibitor of an epigenetic regulator is selected from a bromoterminal and exoterminal domain inhibitor and a lysine-specific histone demethylase 1A inhibitor.

[0250] In some embodiments, the additional therapeutic agent is an immunotherapeutic agent.

[0251] In some embodiments, the immunotherapeutic agent is R-ICE.

[0252] In some embodiments, the additional therapeutic agent is a JAK inhibitor.

[0253] In some embodiments, the JAK inhibitor is selective for JAK1 and JAK2 relative to JAK3 and TYK2. In some embodiments, the JAK inhibitor is selective for JAK1 relative to JAK2, JAK3, and TYK2. For example, the JAK inhibitor may preferentially inhibit JAK1 relative to one or more of JAK2, JAK3, and TYK2. In some embodiments, the JAK inhibitor preferentially inhibits JAK1 relative to JAK2 (e.g., a JAK1 / JAK2 IC 50 Ratio>1). In some embodiments, the JAK inhibitor is 10-fold more selective for JAK1 than for JAK2. In some embodiments, the JAK inhibitor is about 3-fold, about 5-fold, about 10-fold, about 15-fold, or about 20-fold more selective for JAK1 than for JAK2, as measured by IC at 1 mM ATP. 50 Calculated (see, e.g., U.S. Patent Application 14 / 680,659, filed April 7, 2015, the disclosure of which is incorporated herein by reference in its entirety).

[0254] In some embodiments, the additional therapeutic agent is a JAK1 / JAK2 inhibitor. In some embodiments, the additional therapeutic agent is ruxolitinib (eg, ruxolitinib phosphate).

[0255] In some embodiments, the additional therapeutic agent is a JAK inhibitor selected from the group provided in the following table. The compounds provided in the table are selective JAK1 inhibitors (selective over JAK2, JAK3 and TYK2). IC 200 μg / mL obtained by the method of assay A described in U.S. Patent Application 14 / 680,659 at 1 mM ATP 50 Shown in Table A.

[0256] Table A

[0257]

[0258]

[0259]

[0260]

[0261]

[0262]

[0263]

[0264]

[0265] + means <10 nM (for assay conditions, see U.S. Patent Application 14 / 680,659, Example A)

[0266] ++ means ≤100 nM (for assay conditions, see U.S. Patent Application 14 / 680,659, Example A)

[0267] +++ means ≤ 300 nM (for assay conditions, see U.S. Patent Application 14 / 680,659, Example A)

[0268] a Data for enantiomer 1

[0269] b Data for Enantiomer 2

[0270] In some embodiments, the JAK inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile (i.e., itacitinib) or a pharmaceutically acceptable salt thereof.

[0271] In some embodiments, the JAK inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile adipate.

[0272] In some embodiments, the JAK inhibitor is 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide or a pharmaceutically acceptable salt thereof.

[0273] In some embodiments, the JAK inhibitor is selected from (R)-3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propionitrile, (R)-3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propionitrile, (R)-4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2, 3-[(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile, (R)-4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile or (R)-4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile, (S)-3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propionitrile, (S)-3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propionitrile, (S)-4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol -1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile, (S)-4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile, (S)-4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butyronitrile; and pharmaceutically acceptable salts of any of the foregoing.

[0274] In some embodiments, the compounds of Table A are prepared by the synthetic procedures described in the following patent publications: U.S. Patent Publication 2010 / 0298334, filed May 21, 2010, U.S. Patent Publication 2011 / 0059951, filed August 31, 2010, U.S. Patent Publication 2011 / 0224190, filed March 9, 2011, U.S. Patent Publication 2012 / 0149681, filed November 18, 2011, U.S. Patent Publication 2012 / 0149682, filed November 18, 2011, U.S. Patent Publication 2013 / 0018034, filed June 19, 2012, U.S. Patent Publication 2013 / 0045963, filed August 17, 2012, and U.S. Patent Publication 2014 / 0005166, filed May 17, 2013, each of which is incorporated herein by reference in its entirety.

[0275] In some embodiments, the JAK inhibitor is selected from compounds in the following patent publications: U.S. Patent Publication 2010 / 0298334, filed May 21, 2010, U.S. Patent Publication 2011 / 0059951, filed August 31, 2010, U.S. Patent Publication 2011 / 0224190, filed March 9, 2011, U.S. Patent Publication 2012 / 0149681, filed November 18, 2011, U.S. Patent Publication 2012 / 0149682, filed November 18, 2011, U.S. Patent Publication 2013 / 0018034, filed June 19, 2012, U.S. Patent Publication 2013 / 0045963, filed August 17, 2012, and U.S. Patent Publication 2014 / 0005166, filed May 17, 2013, each of which is incorporated herein by reference in its entirety.

[0276] In some embodiments, the PI3Kδ inhibitor and the additional therapeutic agent are administered simultaneously.

[0277] In some embodiments, the PI3Kδ inhibitor and the additional therapeutic agent are administered sequentially.

[0278] Pharmaceutical preparations

[0279] When used as a drug, the compound used in the method of the present disclosure can be administered in the form of a pharmaceutical composition. These compositions can be prepared in a manner well known in the medical field, and can be administered by a variety of routes depending on whether local treatment or systemic treatment is desired and the area to be treated. Administration can be topical (including transdermal, epidermal, ocular and applied to mucous membranes, including intranasal, vaginal and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizers; intratracheal or intranasal), oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial, such as intrathecal or intraventricular administration. Parenteral administration can be in the form of a single bolus dosage form, or can be achieved, for example, by a continuous infusion pump. Pharmaceutical compositions and preparations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional drug carriers, aqueous, powder or oily bases, thickeners, etc. may be necessary or desired.

[0280] The present disclosure also includes pharmaceutical compositions, which contain a compound of the present disclosure or a pharmaceutically acceptable salt thereof as an active ingredient, combined with one or more pharmaceutically acceptable carriers (excipients). In some embodiments, the composition is suitable for topical application. When preparing the composition of the present disclosure, the active ingredient is usually mixed with an excipient, diluted with an excipient or encapsulated in a carrier in the form of, for example, a capsule, a sachet, paper or other container. When an excipient acts as a diluent, it can be a solid, semisolid or liquid material, used as a vehicle, carrier or medium for the active ingredient. Therefore, the composition can be in the following forms: tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solids or in liquid media), ointments containing, for example, up to 10% by weight of active compounds, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders.

[0281] When preparing a formulation, the active compound may be ground to provide an appropriate particle size before combining with other ingredients. If the active compound is substantially insoluble, it may be ground to a particle size of less than 200 mesh. If the active compound is substantially soluble in water, the particle size may be adjusted by grinding to provide a substantially uniform distribution in the formulation, e.g., about 40 mesh.

[0282] The compounds of the present disclosure may be milled using known milling procedures such as wet milling to obtain particle sizes suitable for tablet formation and other formulation types. Finely dispersed (nanoparticulate) formulations of the compounds of the present disclosure may be prepared by processes known in the art, for example see International Application WO 2002 / 000196.

[0283] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup and methylcellulose. The formulation may additionally include: lubricants such as talc, magnesium stearate and mineral oil; wetting agents; emulsifiers and suspending agents; preservatives such as methyl hydroxy-benzoate and propyl hydroxy-benzoate; sweeteners; and flavoring agents. The compositions of the present disclosure can be formulated to provide rapid, sustained or delayed release of the active ingredient after administration to a patient using procedures known in the art.

[0284] The compositions may be formulated in unit dosage form. The term "unit dosage form" refers to physically discrete units suitable as unit dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient.

[0285] To prepare solid compositions, such as tablets, the main active ingredient is mixed with a pharmaceutical excipient to form a solid preformulated composition containing a homogeneous mixture of the disclosed compounds. When these preformulated compositions are referred to as homogeneous, the active ingredient is generally evenly dispersed in the composition so that the composition can be easily subdivided into equally effective unit dosage forms, such as tablets, pills, and capsules. This solid preformulation is then subdivided into unit dosage forms of the above types.

[0286] Tablets or pills of the present disclosure can be coated with coatings or otherwise compounded to produce dosage forms that provide the advantage of extended action. For example, tablets or pills can include an inner dose and an outer dose component, the latter being in the form of an outer inclusion body that coats the former. The two components can be separated by an enteric layer that is used to resist disintegration in the stomach and allows the inner component to enter the duodenum intact or delay release. A variety of materials can be used for such enteric layers or coatings, including a variety of polymeric acids and a mixture of polymeric acids and materials such as shellac, hexadecanol, and cellulose acetate.

[0287] Liquid forms in which the compounds and compositions of the present disclosure may be incorporated for administration orally or by injection include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and emulsions flavored with edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[0288] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, and powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, the composition is administered by oral or nasal respiratory route to achieve local or systemic effects. The composition can be atomized using an inert gas. The atomized solution can be breathed directly from the atomizing device, or the atomizing device can be connected to a mask, curtain, or intermittent positive pressure breathing machine. Solutions, suspensions, or powder compositions can be administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0289] Topical preparations can contain one or more conventional carriers. In some embodiments, ointments can contain water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white vaseline, etc. The carrier composition of the cream can be based on a combination of water and glycerol and one or more other components (for example, glyceryl monostearate, PEG-glyceryl monostearate and hexadecearyl alcohol). Isopropyl alcohol and water can be used, suitably combined with other components (such as glycerol, hydroxyethyl cellulose, etc.) to prepare gels. In some embodiments, topical preparations contain at least about 0.1% by weight, at least about 0.25% by weight, at least about 0.5% by weight, at least about 1% by weight, at least about 2% by weight or at least about 5% by weight of the compound of the present disclosure. Topical preparations can be suitably packaged in, for example, a 100g tube, which is optionally associated with the instructions for treating selected indications (for example, psoriasis or other skin conditions).

[0290] The amount of the compound or composition administered to a patient will vary depending on the agent, the purpose of administration (such as prevention or therapy), the patient's state, the mode of administration, etc. In therapeutic applications, the composition can be administered to a patient already suffering from the disease in an amount sufficient to cure or at least partially arrest the symptoms of the disease and its complications. The effective dose will depend on the disease condition being treated, as well as the judgment of the attending clinician based on factors such as the severity of the disease, the patient's age, weight, and general condition.

[0291] The composition administered to the patient can be in the form of the above-mentioned pharmaceutical composition. These compositions can be sterilized by conventional sterilization techniques, or can be aseptically filtered. The aqueous solution can be packaged for use as is, or lyophilized, and the lyophilized preparation is combined with a sterile aqueous carrier before administration. The pH of the compound preparation will generally be between 3 and 11, more preferably 5 to 9 and most preferably 7 to 8. It should be understood that the use of some of the aforementioned excipients, carriers or stabilizers will result in the formation of pharmaceutical salts.

[0292] The therapeutic dose of the disclosed compounds can vary according to, for example, the specific use to be achieved in the treatment, the mode of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The ratio or concentration of the disclosed compounds in the pharmaceutical composition can vary according to many factors, including dosage, chemical characteristics (e.g., hydrophobicity), and route of administration. For example, the disclosed compounds can be provided in a physiological buffered aqueous solution containing about 0.1 w / v% (weight / volume %) to about 10 w / v% of the compound for parenteral administration.

[0293] The compositions of the present disclosure may also include one or more additional agents, such as chemotherapeutic agents, steroids, anti-inflammatory compounds, or immunosuppressive agents, examples of which are listed herein.

[0294] Labeled compounds and assays

[0295] The methods of the present disclosure also include the use of isotopically labeled compounds of the present disclosure. An "isotope" or "radiolabeled" compound is a compound of the present disclosure in which one or more atoms are replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number normally found in nature (i.e., naturally occurring). Suitable radionuclides that can be incorporated into the compounds of the present disclosure include, but are not limited to 2 H (also written as D, i.e. deuterium), 3 H (also written as T, i.e. tritium), 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 18 F. 35 S. 36 Cl, 82 Br, 75 Br, 76 Br, 77 Br, 123 I. 124 I. 125 I and 131 I. For example, one or more hydrogen atoms in the compounds of the present disclosure may be replaced by a deuterium atom (e.g., one or more hydrogen atoms of an alkyl group of a compound described herein may be optionally replaced by a deuterium atom, such as -CD 3 Substitute -CH 3 ).

[0296] One or more constituent atoms of the compounds provided herein can be replaced or substituted with isotopes of the atoms in natural or non-natural abundance. In some embodiments, the compound includes at least one deuterium atom. In some embodiments, the compound includes two or more deuterium atoms. In some embodiments, the compound includes 1-2, 1-3, 1-4, 1-5 or 1-6 deuterium atoms. In some embodiments, all hydrogen atoms in the compound can be replaced or substituted with deuterium atoms.

[0297] In some embodiments, 1, 2, 3, 4, 5, 6, 7, or 8 hydrogen atoms attached to carbon atoms of the compounds described herein are optionally replaced with deuterium atoms.

[0298] Synthetic methods for including isotopes in organic compounds are known in the art (Deuterium Labeling in Organic Chemistry, Alan F. Thomas (New York, NY, Appleton-Century-Crofts, 1971; The Renaissance of H / D Exchange, Jens Atzrodt, Volker Derdau, Thorsten Fey and Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765; The Organic Chemistry of Isotopic Labelling, James R. Hanson, Royal Society of Chemistry, 2011). Isotopically labeled compounds can be used in a variety of studies such as NMR spectroscopy, metabolic experiments and / or assays.

[0299] Substitution with heavier isotopes such as deuterium may provide certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and may therefore be preferred in some circumstances. (See, e.g., A. Kerekes et al. J. Med. Chem. 2011, 54, 201-210; R. Xu et al. J. Label Compd. Radiopharm. 2015, 58, 308-312). In particular, substitution at one or more metabolic sites may provide one or more therapeutic advantages.

[0300] The radionuclide incorporated into the radiolabeled compounds of the invention will depend on the specific application of the radiolabeled compound. For example, for in vitro PI3K labeling and competition assays, incorporation of 3 H. 14 C. 82Br, 125 I. 131 I or 35 Compounds of S may be useful. For radioactive imaging applications, 11 C. 18 F. 125 I. 123 I. 124 I. 131 I. 75 Br, 76 Br or 77 Br can be useful.

[0301] It is understood that a "radiolabeled" or "labeled compound" is a compound into which at least one radionuclide has been incorporated. In some embodiments, the radionuclide is selected from 3 H. 14 C. 125 I. 35 S and 82 The group composed of Br.

[0302] The present disclosure may also include synthetic methods for incorporating radioisotopes into compounds of the present disclosure.Synthetic methods for incorporating radioisotopes into organic compounds are well known in the art, and one of ordinary skill in the art will readily recognize methods applicable to the compounds of the present disclosure.

[0303] Reagent test kit

[0304] The present disclosure also includes a pharmaceutical kit suitable for, for example, treating or preventing a PI3K-related disease or condition described herein, comprising one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a compound of the present disclosure. Such kits may also include (if desired) one or more of various conventional pharmaceutical kit components, such as a container with one or more pharmaceutically acceptable carriers, additional containers, etc., as will be readily apparent to those skilled in the art. Instructions in the form of inserts or labels may also be included in the kit, indicating the amount of the component to be administered, guidelines for administration, and / or guidelines for mixing the components. In certain embodiments, the instructions include methods of the present disclosure, including prescribed doses or dosing regimens of the present disclosure.

[0305] The present invention will be described in more detail by means of specific examples. The following examples are provided to achieve illustrative purposes, and are not intended to limit the present invention in any way. Those skilled in the art will readily recognize that a variety of non-critical parameters that can be changed or modified to produce substantially the same results. It should be understood that some features of the present invention described in the context of the respective embodiments for clarity may also be provided in combination in a single embodiment. On the contrary, the various features of the present invention described in the context of a single embodiment for brevity may also be provided separately or in any suitable sub-combination.

[0306] Example

[0307] Examples 1A-1D. Diastereomers of 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one

[0308]

[0309] Step 1. 1-(5-chloro-2-ethoxy-3-iodo-4-methylphenyl)ethanol

[0310]

[0311] A solution of 1-(5-chloro-2-ethoxy-4-fluoro-3-iodophenyl)ethanone (20.0 g, 58.4 mmol; see Example 212, Step 1 of U.S. Patent 9,199,982) and 1,2-ethylene glycol (6.5 mL, 120 mmol) in toluene (190 mL) was treated with p-toluenesulfonic acid monohydrate (1.1 g, 5.8 mmol). The flask was equipped with a Dean-Stark trap filled with sieves and refluxed for 3 h. The reaction mixture was cooled and added to an ice-cold saturated sodium bicarbonate solution (250 mL) and extracted with ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate, filtered, and concentrated to a crude orange oil. The crude material was purified by flash column chromatography using hexanes containing ethyl acetate (0%-20%) to give the desired product (22 g, 99%). 12 H 14 ClFIO 3 LCMS: (M+H) + :m / z=387.0; Found: 386.9.

[0312] Step 2. Ethyl (2E)-3-[3-chloro-6-ethoxy-2-fluoro-5-(2-methyl-1,3-dioxolan-2-yl)phenyl]acrylate

[0313]

[0314] A mixture of 2-(5-chloro-2-ethoxy-4-fluoro-3-iodophenyl)-2-methyl-1,3-dioxolane (22 g, 58 mmol) (from step 1), ethyl (2E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)acrylate (16 mL, 70 mmol) and potassium carbonate (24 g, 170 mmol) in 1,4-dioxane (230 mL) and water (110 mL) was degassed with nitrogen for 10 minutes. The reaction mixture was treated with [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II), complexed with dichloromethane (1:1) (2.4 g, 2.9 mmol), degassed with nitrogen for 10 minutes, and heated at 80°C for 2 h. The reaction mixture was filtered through celite and washed with ethyl acetate (300 mL). The filtrate was poured into water (400 mL). The aqueous layer was separated and extracted with ethyl acetate (300 mL). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated into a crude brown solid. The crude material was purified by flash column chromatography using hexane (0%-30%) containing ethyl acetate to give the desired product (20 g, 96%). 1 H NMR (400 MHz, CDCl 3 )δ7.74(d,J=16.5Hz,1H),7.56(d,J=8.6Hz,1H),6.70(dd,J=16.5,0.9Hz,1H),4.26(q,J=7.1Hz,2H),4.10–3. 99(m,2H),3.91(q,J=7.0Hz,2H),3.87–3.76(m,2H),1.73(s,3H),1.44(t,J=7.0Hz,3H),1.33(t,J=7.1Hz,3H). C 17 H 21 ClFO 5 LCMS: (M+H) + :m / z=359.1; measured value: 359.1.

[0315] Step 3. Ethyl 3-[3-chloro-6-ethoxy-2-fluoro-5-(2-methyl-1,3-dioxolan-2-yl)phenyl]-4-nitrobutyrate

[0316]

[0317] A solution of (2E)-3-[3-chloro-6-ethoxy-2-fluoro-5-(2-methyl-1,3-dioxolan-2-yl\)phenyl]ethyl acrylate (10 g, 28 mmol) (from step 2) in nitromethane (100 mL) was treated with 1,8-diazabicyclo[5.4.0]11-7-ene (4.6 mL, 31 mmol) and stirred at 60 ° C for 15 h. The reaction mixture was poured into water (400 mL) and extracted with ethyl acetate (2 x 300 mL). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated to a crude orange oil. The crude material was purified by flash column chromatography using hexane (0%-30%) containing ethyl acetate to give the desired product (10.4 g, 89%) as a mixture of enantiomers. 1 H NMR (400 MHz, CDCl 3 )δ7.52(d,J=9.1Hz,1H),4.82(ddd,J=12.5,7.6,1.4Hz,1H),4.68(dd,J=12.5,7.2Hz,1H),4.54–4.40(m,1H),4.15–3.90(m,6H),3.89–3 .75(m,2H),2.85(ddd,J=16.0,8.6,1.4Hz,1H),2.73(dd,J=16.1,6.2Hz,1H),1.70(s,3H),1.47(t,J=7.0Hz,3H),1.21(t,J=7.1Hz,3H). C 18 H 24 ClFNO 7 LCMS: (M+H) + :m / z=420.1; measured value: 420.1.

[0318] Step 4. Enantiomers of 4-[3-chloro-6-ethoxy-2-fluoro-5-(2-methyl-1,3-dioxolan-2-yl)phenyl]pyrrolidin-2-one

[0319]

[0320] A suspension of 3-[3-chloro-6-ethoxy-2-fluoro-5-(2-methyl-1,3-dioxolane-2-yl)phenyl]-4-nitrobutyric acid ethyl ester (1.0 g, 2.4 mmol) (from step 3) in ethanol (16 mL) was heated to dissolve the solid. The solution was cooled back to ambient temperature, degassed with nitrogen, and treated with a slurry of 2800 Raney Nickel in water (1.5 mL). The reaction mixture was degassed again with nitrogen and hydrogenated with a hydrogen balloon for 3 h. The reaction mixture was filtered through diatomaceous earth and concentrated to give an intermediate amino ester (0.93 g, 100%). The intermediate amino ester was dissolved in toluene (12 mL) and heated at 110 ° C for 12 h. The reaction mixture was cooled to ambient temperature, at which time solids precipitated from the solution. The mixture was cooled to 0°C, stirred for 30 minutes, filtered, washed with cold toluene, and dried to give the desired product as a mixture of enantiomers (0.61 g, 75%). 16 H 20 ClFNO 4 LCMS: (M+H) + : m / z = 344.1; found: 344.1. The mixture of enantiomers was separated by chiral HPLC to give the individual enantiomers as peak 1 and peak 2 (RT = 5.39 min and 7.01 min, respectively; Phenomenex Lux Cellulose C-1, 21.2 x 250 mm, 5 micron particle size, eluted with 20% ethanol in hexanes at 18 mL / min).

[0321] Step 5. Enantiomers of 4-(3-acetyl-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one

[0322]

[0323] The separated enantiomers from step 4 were each processed individually to the final compound. A solution of 4-[3-chloro-6-ethoxy-2-fluoro-5-(2-methyl-1,3-dioxolan-2-yl)phenyl]pyrrolidin-2-one (1.7 g, 5.0 mmol) (from step 4) in methanol (17 mL) was treated dropwise with water (11 mL, 69 mmol) containing 6.0 M hydrogen chloride and stirred at 20 ° C for 30 minutes. The reaction mixture was added dropwise to an ice-cold saturated sodium bicarbonate solution (75 ml) and extracted with ethyl acetate (2x100 ml). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated to give the desired product [from peak 1 (1.5 g, 99%); from peak 2 (1.5 g, 99%)], which was used without further purification. From peak 1: 1 HNMR (400 MHz, DMSO-d6 )δ7.84(s,1H),7.70(d,J=8.6Hz,1H),4.16–3.99(m,1H),3.83(q,J=7.0Hz,2H),3.65–3.54( m,1H),3.30–3.23(m,1H),2.55(s,3H),2.33(dd,J=16.8,8.4Hz,1H),1.30(t,J=7.0Hz,3H). C 14 H 16 ClFNO 3 LCMS: (M+H) + :m / z=300.1; Found: 300.0. From Peak 2: 1 H NMR (400 MHz, DMSO-d 6 )δ7.84(s,1H),7.70(d,J=8.6Hz,1H),4.13–4.00(m,1H),3.87–3.77(m,2H),3.65–3.55(m,1H ), 3.31–3.23(m,1H),2.55(s,3H),2.32(ddd,J=16.9,8.4,1.6Hz,1H),1.30(t,J=7.0Hz,3H). C 14 H 16 ClFNO 3 LCMS: (M+H) + :m / z=300.1; measured value: 300.1.

[0324] Step 6. Diastereomers of 4-[3-chloro-6-ethoxy-2-fluoro-5-(1-hydroxyethyl)phenyl]pyrrolidin-2-one

[0325]

[0326] The enantiomers from step 5 are processed into final products separately. At 0 ° C, a solution of 4-(3-acetyl 5-chloro-2-ethoxy 6-fluorophenyl) pyrrolidin-2-one (0.402g, 1.34mmol) (from step 5) in anhydrous methanol (6.7mL) is treated with sodium tetrahydroborate (0.10g, 2.7mmol) under a nitrogen atmosphere and stirred for 30 minutes at 0 ° C. The reaction mixture is quenched with water at 0 ° C and poured into water (50mL) / ethyl acetate (100mL) while stirring. The mixture is warmed to ambient temperature and the water layer is separated and extracted with ethyl acetate (50mL) again. The combined organic extracts are washed with brine, dried over sodium sulfate, filtered, and concentrated to give a white foam. The crude material was purified by flash column chromatography using acetonitrile (containing 7% methanol) in dichloromethane (0%-100%) to give the desired product as a mixture of diastereomers [from Peak 1 (0.40 g, 99%); from Peak 2 (0.40 g, 99%)]. From Peak 1: C 14 H 18 ClFNO 3 LCMS: (M+H) + :m / z=302.1; Found: 302.0. From Peak 2: C 14 H 18 ClFNO 3 LCMS: (M+H) + :m / z=302.1; measured value: 302.1.

[0327] Step 7. Diastereomers of 4-[3-chloro-5-(1-chloroethyl)-6-ethoxy-2-fluorophenyl]pyrrolidin-2-one

[0328]

[0329] The mixture of diastereomers from step 6 was processed individually to the final product. A solution of 4-[3-chloro-6-ethoxy-2-fluoro-5-(1-hydroxyethyl)phenyl]pyrrolidin-2-one (0.41 g, 1.4 mmol) (from step 6) in dichloromethane (12 mL) was treated dropwise with N,N-dimethylformamide (0.011 mL, 0.14 mmol) and thionyl chloride (0.21 mL, 2.9 mmol) and stirred at 20°C for 30 minutes. The reaction mixture was added dropwise to an ice-cold saturated sodium bicarbonate solution and extracted with dichloromethane. The organic layer was separated and washed with brine, dried over sodium sulfate, filtered, and concentrated to give the desired product [from peak 1 (0.38 g, 87%); from peak 2 (0.39 g, 89%)] and 17-18% of styrene formed by chloride elimination. These mixtures were used without further purification. From peak 1: C14 H 17 Cl 2 FNO 2 LCMS: (M+H) + :m / z=320.1; Found: 320.0. From Peak 2: C 14 H 17 Cl 2 FNO 2 LCMS: (M+H) + :m / z=320.1; measured value: 320.0.

[0330] Step 8. Diastereomers of 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one

[0331]

[0332] The mixture of diastereomers from step 7 was processed individually to the final product. A mixture of 4-[3-chloro-5-(1-chloroethyl)-6-ethoxy-2-fluorophenyl]pyrrolidin-2-one (0.36 g, 1.1 mmol) (from step 7), 3-methyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (0.19 g, 1.3 mmol), cesium carbonate (0.54 g, 1.7 mmol) and potassium iodide (18 mg, 0.11 mmol) in N,N-dimethylformamide (7.4 mL) was heated at 100° C. for 4.5 h. The reaction mixture was poured into water (30 ml) and washed with ethyl acetate (3×10 % ethyl acetate). 50mL) was extracted to obtain the diastereoisomers ((S)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidine -2-one; (S)-4-(3-((R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one; and (R)-4-(3-((R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one). The mixture of diastereomers was purified by preparative LCMS (XBridge C18 column, eluted with a gradient of acetonitrile / water containing 0.1% ammonium hydroxide at a flow rate of 60 mL / min) to give the desired product [Peak A (Example 1A) (0.13 g, 54%) and Peak B (Example 1B) (0.11 g, 46%) were separated from Peak 1; Peak A (Example 1C) (0.15 g, 63%) and Peak B (Example 1D) (0.14 g, 55%) were separated from Peak 2].

[0333] Example 1B: 1 H NMR (300 MHz, DMSO-d 6)δ8.12(s,1H),7.82(s,1H),7.52(d,J=8.5Hz,1H),7.30(br s,1H),6.23(q,J=7.0Hz,1H),4.05–3.90(m,1H),3.88–3.78(m,2H),3.63–3.53(m,1H),3.29 –3.20(m,1H),2.54(s,3H),2.38–2.21(m,1H),1.70(d,J=7.1Hz,3H),1.39(t,J=6.9Hz,3H). C 20 H 23 CIF 6 O 2 LCMS: (M+H) + :m / z=433.2; Found: 433.1. Example 1C: 1 H NMR (500 MHz, DMSO-d 6 )δ8.12(s,1H),7.77(s,1H),7.53(d,J=8.5Hz,1H),7.26(br s,2H),6.24(q,J=7.0Hz,1H),4.04–3.94(m,1H),3.93–3.85(m,1H),3.84–3.77(m,1H),3.61–3.53(m,1H), 3.27–3.22(m,1H),2.54(s,3H),2.30(dd,J=18.1,8.6Hz,1H),1.71(d,J=7.1Hz,3H),1.40(t,J=6.9Hz,3H). C 20 H 23 CIF 6 O 2 LCMS: (M+H) + :m / z=433.2; measured value: 433.1.

[0334] Examples 2A-2D. Diastereomers of 5-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}-1,3-oxazolidin-2-one

[0335]

[0336] Step 1: tert-Butyl [2-(3-acetyl-5-chloro-2-ethoxy-6-fluorophenyl)-2-hydroxyethyl]carbamate

[0337]

[0338] The aqueous solution (10mL) of 0.2M osmium tetroxide is added to a solution of [(4-chlorobenzoyl)oxy] tert-butyl carbamate (Lawrence Harris, J.Org.Chem, 2011, 76, 358-372) (19g, 70mmol) in acetonitrile (210mL) and stirred for 10 minutes. 1-(5-chloro-2-ethoxy-4-fluoro-3-vinylphenyl) ethyl ketone (11.2g, 46mmol) (see Example 353, step 1 of U.S. Patent 9,199,982) in the form of a solution in acetonitrile (210mL) is added to the carbamate solution, followed by addition of water (50mL) and stirring at room temperature for 3 hours. The reaction is quenched with a saturated 10M potassium disulfite aqueous solution (240mL) and stirred for 5 minutes. Water is added and the reaction mixture is extracted with ethyl acetate. The extract is washed with saturated sodium bicarbonate solution, brine and dried over sodium sulfate, filtered and evaporated. Purification on silica gel using ethyl acetate in hexanes (0-100%) gave the desired compound as a racemic mixture, 16.6 g, 95%. 17 H 23 ClFNO 5 LCMS calculated value of Na: (M+Na) + :m / z=398.1; Found: 398.0.

[0339] Step 2: 5-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}-1,3-oxazolidin-2-one

[0340]

[0341] The desired single enantiomer (Peak 3) was prepared using the same procedure as Example 353, Steps 8-12 of U.S. Pat. No. 9,199,982, except that the intermediate from Step 1 in this example was a racemate and therefore the final separation of the four diastereomers was performed in Step 12. Chiral purification on a Phenomenex Lux Cellulose C-4, 21 x 250 mm (Chiral Technologies, 5 micron particle size) using 30% ethanol in hexanes at a flow rate of 18 mL / min afforded Peak 1: Example 2A (single enantiomer) (Retention time = 12.7 minutes), Peak 2: Example 2B (single enantiomer) (Retention time = 14.2 minutes), Peak 3: Example 2C (single enantiomer) (Retention time = 20.3 minutes), and Peak 4: Example 2D (single enantiomer) (Retention time = 28.9 minutes); the most active enantiomer was Peak 3. 19H 21 CIF 6 O 3 LCMS calculated value: (M+H) + :m / z=435.1; Found: 435.1. 1 H NMR (500 MHz, DMSO-d 6 ): δ8.15(s,1H),7.81(s,1H),7.71(d,1H),7.26(bs,1H),6.23(m,1H),5.8 4(t,1H),3.92(m,1H),3.83(m,1H),2.52(s,3H),1.75(d,3H),1.40(m,3H).

[0342] Example A1: PI3K enzyme assay

[0343] The PI3 kinase luminescent assay kit was purchased from Echelon Biosciences (Salt Lake City, UT), and the kit includes the lipid kinase substrate D-myo-phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) D(+)-sn-1,2-di-O-octanoylglycerol, 3-O-phosphate-linked (PIP2), biotinylated I(1,3,4,5)P4, and PI(3,4,5)P3 detection protein. AlphaScreen including donor beads and acceptor beads TM GST detection kit was purchased from PerkinElmer LifeSciences (Waltham, MA). PI3Kδ (p110δ / p85α) was purchased from Millipore (Bedford, MA). ATP, MgCl 2 , DTT, EDTA, HEPES and CHAPS were purchased from Sigma-Aldrich (St. Louis, MO).

[0344] AlphaScreen for PI3Kδ TM Determination

[0345] Kinase reactions were performed in 384-well REMP plates from Thermo Fisher Scientific in a final volume of 40 μL. Inhibitors were first serially diluted in DMSO and added to the wells, followed by the addition of other reaction components. The final concentration of DMSO in the assay was 2%. The cells were incubated in 50 mM HEPES (pH 7.4), 5 mM MgCl 2PI3K assays were performed in 50 mM NaCl, 5 mM DTT, and 0.04% CHAPS. The reaction was initiated by adding ATP, and the final reaction mixture consisting of 20 μM PIP2, 20 μM ATP, 1.2 nM PI3Kδ was incubated for 20 minutes. 10 μL of the reaction mixture was then transferred to 5 μL of 50 nM biotinylated I(1,3,4,5)P4 in the following quenching buffer: 50 mM HEPES pH 7.4, 150 mM NaCl, 10 mM EDTA, 5 mM DTT, 0.1% Tween-20, followed by the addition of 10 μL of AlphaScreen suspended in quenching buffer containing 25 nM PI(3,4,5)P3 detection protein. TM Donor beads and acceptor beads. The final concentration of both donor beads and acceptor beads was 20 mg / ml. After sealing the plate, the plate was incubated at room temperature in a dark location for 2 hours. Product activity was determined on a Fusion-α microplate reader (Perkin-Elmer). IC was performed by fitting a curve of the control activity percentage relative to the logarithm of the inhibitor concentration using GraphPad Prism 3.0 software. 50 Sure.

[0346] Example A2: PI3K enzyme assay

[0347] Material

[0348] The lipid kinase substrate phosphoinositide-4,5-bisphosphate (PIP2) was purchased from Echelon Biosciences (Salt Lake City, UT). PI3K isoforms α, β, δ, and γ were purchased from Millipore (Bedford, MA). ATP, MgCl 2 , DTT, EDTA, MOPS, and CHAPS were purchased from Sigma-Aldrich (St. Louis, MO).

[0349] Kinase reactions were performed in clear-bottom 96-well plates from Thermo Fisher Scientific in a final volume of 24 μL. Inhibitors were first serially diluted in DMSO and added to the wells, followed by the addition of other reaction components. The final concentration of DMSO in the assay was 0.5%. The cells were incubated in 20 mM MOPS (pH 6.7), 10 mM MgCl at room temperature. 2 PI3K assays were performed in 50 μM 5 mM DTT and 0.03% CHAPS. Reaction mixtures containing 50 μM PIP2, kinase and different concentrations of inhibitors were prepared. 33The reaction was initiated by adding 100 μM of ATP to a final concentration of 1000 μM of ATP. The final concentrations of PI3K isoforms α, β, δ and γ in the assay were 1.3, 9.4, 2.9 and 10.8 nM, respectively. The reaction was incubated for 180 minutes and terminated by adding 100 μL of 1M potassium phosphate (pH 8.0), 30 mM EDTA quenching buffer. Then 100 μL of reaction solution aliquots were transferred to 96-well Millipore MultiScreen IP 0.45 μm PVDF filter plates (filter plates were pre-wetted with 200 μL 100% ethanol, distilled water and 1M potassium phosphate (pH 8.0), respectively). The filter plate was aspirated under vacuum on a Millipore Manifold and washed with 18 × 200 μL of a wash buffer containing 1M potassium phosphate (pH 8.0) and 1 mM ATP. After drying by aspiration and wicking, the plates were air-dried overnight in an incubator at 37°C. A Packard TopCount adapter (Millipore) was then connected to the plate, followed by the addition of 120 μL of Microscint 20 scintillation cocktail (PerkinElmer) to each well. After sealing the plate, the radioactivity of the product was determined by scintillation counting on a Topcount (Perkin-Elmer). IC was determined by fitting a curve of the percentage of control activity versus the logarithm of the inhibitor concentration using GraphPad Prism 3.0 software. 50 Sure.

[0350] Example A3: PI3Kδ scintillation proximity assay

[0351] Material

[0352] [γ- 33 P]ATP (10 mCi / mL) was purchased from Perkin–Elmer (Waltham, MA). The lipid kinase substrate D-myo-phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) D(+)-sn-1,2-di-O-octanoylglycerol, 3-O-phosphate-linked (PIP2) (CAS 204858-53-7) was purchased from Echelon Biosciences (Salt Lake City, UT). PI3Kδ (p110δ / p85α) was purchased from Millipore (Bedford, MA). ATP, MgCl 2DTT, EDTA, MOPS and CHAPS were purchased from Sigma-Aldrich (St. Louis, MO). Wheat Germ Agglutinin (WGA) YSi SPA scintillation beads were purchased from GE healthcare life sciences (Piscataway, NJ).

[0353] Kinase reactions were performed in polystyrene 384-well matrix white plates from Thermo Fisher Scientific in a final volume of 25 μL. Inhibitors were first serially diluted in DMSO and added to the wells, followed by the addition of other reaction components. The final concentration of DMSO in the assay was 0.5%. The 50 μL lysate was added to the plate in 20 mM MOPS (pH 6.7), 10 mM MgCl at room temperature. 2 PI3K assay was performed in 5 mM DTT and 0.03% CHAPS. The reaction was initiated by adding ATP, and the final reaction mixture consisted of 20 μM PIP2, 20 μM ATP, 0.2 μCi [γ- 33 The reaction was incubated for 210 minutes and terminated by adding 40 μL SPA beads suspended in the following quenching buffer: 150 mM potassium phosphate (pH 8.0), 20% glycerol, 25 mM EDTA, 400 μM ATP. The final concentration of SPA beads was 1.0 mg / mL. After sealing the plate, the plate was shaken overnight at room temperature and centrifuged at 1800 rpm for 10 minutes, and the radioactivity of the product was determined by scintillation counting on a Topcount (Perkin-Elmer). IC was performed by fitting a curve of the percentage of control activity relative to the logarithm of the inhibitor concentration using GraphPad Prism 3.0 software. 50 As determined by Assay A, the IC of the Example 50 The data are given in Table B.

[0354] Table B

[0355] Example No. <![CDATA[PI3KδSPA IC 50 (nM)*]]> 1A +++ 1B + 1C + 1D +++ 2A +++ 2B +++ 2C + 2D +++++

[0356] * Symbols in the column:

[0357] + indicates ≤10nM

[0358] ++ means >10nM to 50nM

[0359] +++ means >50nM to 200nM

[0360] ++++ means >200nM to 500nM

[0361] +++++ means >500nM

[0362] Example B1: B cell proliferation assay

[0363] To obtain B cells, human PBMCs were isolated from peripheral blood of normal drug-free donors by standard density gradient centrifugation on Ficoll-Hypague (GE Healthcare, Piscataway, NJ) and incubated with anti-CD19 microbeads (Miltenyi Biotech, Auburn, CA). B cells were then purified by positive immunosorting using autoMacs (Miltenyi Biotech) according to the manufacturer's instructions.

[0364] Purified B cells (2×10 5 / well / 200 μL) were cultured in RPMI1640, 10% FBS and goat F(ab')2 anti-human IgM (10 μg / ml) (Invitrogen, Carlsbad, CA) in the presence of different amounts of test compounds in 96-well ultra-low binding plates (Corning, Corning, NY) for three days. 3 H]-thymidine (1 μCi / well) (PerkinElmer, Boston, MA) in PBS was added to B cell cultures for an additional 12 h, after which bound radioactivity was isolated by filtration with water through GF / B filters (Packard Bioscience, Meriden, CT) and measured by liquid scintillation counting with a TopCount (Packard Bioscience).

[0365] Example B2: Pfeiffer cell proliferation assay

[0366] Pfeiffer cell line (diffuse large B-cell lymphoma) was purchased from ATCC (Manassas, VA) and maintained in recommended medium (RPMI and 10% FBS). To measure the antiproliferative activity of compounds, Pfeiffer cells were plated with medium (2x10 3 cells / well / 200 μl) into a 96-well ultra-low binding plate (Corning, Corning, NY). After 3-4 days, the cells containing [ 3H]-thymidine (1 μCi / well) (PerkinElmer, Boston, MA) in PBS was added to the cell cultures for an additional 12 h, after which the bound radioactivity was isolated by filtration with water through GF / B filters (Packard Bioscience, Meridenj, CT) and measured by liquid scintillation counting with a TopCount (Packard Bioscience). IC of selected compounds 50 The data are given in Table C.

[0367] Table C

[0368] Example No. <![CDATA[Pfeiffer IC 50 (nM)*]]> 1B + 1C + 2C +

[0369] * Symbols in the column:

[0370] + indicates ≤10nM

[0371] ++ means >10nM to 50nM

[0372] Example C: Akt phosphorylation assay

[0373] Ramos cells (B lymphocytes from Burkitt's lymphoma) were obtained from ATCC (Manassas, VA) and maintained in RPMI1640 and 10% FBS. 7 Cells / tube / 3mL in RPMI) were incubated with different amounts of test compounds at 37°C for 2 hours and then stimulated with goat F(ab')2 anti-human IgM (5μg / mL) (Invitrogen) in a 37°C water bath for 17 minutes. The stimulated cells were briefly centrifuged at 4°C and whole cell extracts were prepared using 300μL lysis buffer (Cell Signaling Technology, Danvers, MA). The resulting lysate was sonicated and the supernatant was collected. The phosphorylation level of Akt in the supernatant was analyzed by using PathScan Phospho-Akt1 (Ser473) Sandwich ELISA Kit (Cell Signaling Technology) according to the manufacturer's instructions.

[0374] Example D: Pfeiffer Lymphoma Model

[0375] method

[0376] Female SCID mice (5 to 8 weeks old, Charles River Laboratories, Wilmington, MA) were inoculated with 0.2 mL of sterile saline containing 1×107 tumor cells (Pfeiffer, ATCC#CRL-2632, Manassas, VA) and matrigel (BD Biosciences#354234). Inoculation was performed subcutaneously on the flank. Tumor tissue fragments (approximately 3 mm×3 mm) were collected 3 to 6 weeks after cultured cell inoculation and implanted subcutaneously in place of cell inoculation. Tissue fragments were implanted in the form of solid fragments using blunt-tipped forceps. Depending on the size of the tumor, treatment of tumor-bearing mice was started 15 to 25 days after tumor inoculation. Animals were sorted so that the average tumor volume in each group was approximately equal. On the first day of treatment, the minimum average tumor volume in all groups was 150 mm3 and each group consisted of 7 animals. Experimental therapeutic agent Example 347 was administered to mice orally (PO). The frequency of treatment was 2 times a day for a minimum of 14 days to achieve efficacy. The size of subcutaneous tumors was measured 2 to 3 times a week using a digital caliper. Tumor volume was calculated by measuring the tumor in 2 dimensions and using the equation: volume = [length × width 2)] / 2; where the larger value is the length and the smaller value is the width. If multiple tumors are formed, the final volume is the sum of the individual tumors that obey the same equation: for example, 2 tumors; volume = {[L1×(W1)2] / 2}+{[L2 x(W2)2] / 2}. The effect on tumor growth is reported as tumor growth inhibition percentage (TGI%). The TGI percentage is calculated using the equation: (1-(Tx volume / control volume))*100, where the control volume is the volume of the vehicle or untreated tumor on a given day, and the Tx volume is the volume of any treatment group tumor on the same day. ANOVA: single factor test was used to assess the statistical differences between treatment and vehicle control.

[0377] result

[0378] (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one (see Example 1) was evaluated as a single agent in the Pfeiffer human tumor xenograft model of diffuse large B-cell lymphoma, a subtype of NHL. Pfeiffer cancer cells were shown to be sensitive to the antiproliferative effects of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one in vitro. Thus, a tumor model was established based on subcutaneous inoculation of tumor cells into immunocompromised SCID mice and tumor-bearing mice, which received oral doses of vehicle or (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one at 0.3, 1, 3 or 10 mg / kg twice daily for 14 days. (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one treatment inhibited tumor growth by 22%, 24%, 36% and 58% (percent tumor growth inhibition) with increasing doses (see, e.g., U.S. Pat. No. 9,932,341).

[0379] Example E: Dose escalation and dose expansion studies

[0380] In the dose escalation and dose expansion studies described herein, the PK, PD, safety, and efficacy of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride (i.e., Compound 1C) were evaluated in patients with relapsed or refractory B-cell malignancies. The PK characteristics of Compound 1C in the absence of dose-limiting toxicity (DLT) allowed for exceeding the IC at all tested doses throughout the once-daily (QD) dosing interval. 90 In addition, the results of the food effect analysis indicate that compound 1C can be administered regardless of meal time. The near-maximal inhibition of pAKT in the ex vivo PD assay is consistent with the exposure data. The results of PI3Kδ monotherapy and combination therapy are discussed in Examples FH below.

[0381] Study Design and Patients

[0382] The dose escalation and dose expansion study was conducted in multiple parts: dose escalation of compound 1C monotherapy (part 1) followed by cohort expansion (part 3); compound 1C + itatinib dose escalation (part 2) followed by cohort expansion (part 3); and compound 1C + R-ICE (rituximab plus ifosfamide, carboplatin, and etoposide) dose evaluation and expansion (part 6) (NCT02018861). Eligible patients (age ≥18 years) had a diagnosis of lymphoid malignancies of B-cell origin, including indolent or aggressive B-cell NHL malignancies, transformed NHL histology, and Hodgkin lymphoma (HL), who had relapsed or were refractory to previous standard therapy. Patients with Burkitt's lymphoma and precursor B lymphoblastic leukemia / lymphoma were excluded. Eligible patients had an Eastern Cooperative Oncology Group performance status (ECOGPS) ≤1 (dose escalation) or ≤2 (dose expansion); had adequate cardiac, hepatic, and renal function; had a life expectancy of ≥12 weeks; had received ≥1 prior treatment regimen; and were unresponsive to or not a candidate for hematopoietic (H)SCT or other potential curative therapies.

[0383] Patients will be excluded if they have a history of: untreated, symptomatic, or unstable brain metastases; spinal cord compression; lymphoma involving the central nervous system (allowed in Compound 1C monotherapy expansion cohort A [B-cell malignancies] and Compound 1C + itatinib combination expansion cohort); or allogeneic hematopoietic stem cell transplant (HSCT) within 6 months or autologous HSCT within 3 months of enrollment. Prior treatment with PI3Kδ inhibitors was excluded from the Compound 1C monotherapy cohort, and prior JAK inhibitors were excluded from the Compound 1C + itatinib combination therapy cohort, unless approved by the medical supervisor.

[0384] 72 patients were enrolled and treated with Compound 1C monotherapy (median age, 66 [range, 30–89] years) (Table 1). The median (range) duration of treatment was 4.0 (0.2-22.7) months. At the time of data cutoff, all but 11 subjects had discontinued study treatment, primarily due to disease progression (n=35 [49%]) and adverse events (AEs) (n=14 [19%]).

[0385] Table 1. Patient demographics and baseline characteristics (ITT)

[0386]

[0387] *Includes American Indian, Alaska Native, Pacific Islander, and Other. These included classical HL (n=9) and nodular lymphocyte-predominant HL (n=1). These included extranodal MZL of MALT type (n=2), nodular MZL (n=4), splenic MZL (n=2), and unknown MZL subtype (n=1). Classical HL (n=2) was included. CLL: chronic lymphocytic leukemia; DLBCL: diffuse large B-cell lymphoma; FL: follicular lymphoma; HL: Hodgkin lymphoma; HSCT: hematopoietic stem cell transplantation; MALT: mucosa-associated lymphoid tissue; MCL: mantle cell lymphoma; MZL: marginal zone lymphoma; R-ICE: rituximab plus ifosfamide, carboplatin, and etoposide; WM: Waldenstrom's macroglobulinemia.

[0388] Drug administration

[0389] For compound 1C monotherapy, compound 1C is orally self-administered once a day (QD). Compound 1C is administered in a fasting state, with the exception that patients join an optional food effect group until the protocol amendment allows administration without regard to food. Treatment continues until unacceptable toxicity or disease progression. During dose escalation (see Table 2C), the initial single patient group is treated with 5mgQD and subsequent groups are recruited using a 3+3 design to identify any dose-limiting toxicity (DLT; defined in Table 2A) during the 21-day observation period. If DLT occurs in >1 of the first 3 patients or in the total group of 6 patients, it is considered to exceed the maximum tolerated dose (MTD) and the next lower tolerable dose level is defined as MTD. All doses up to 45mg QD are well tolerated in the 21-day DLT observation period, and are therefore not identified as MTD. However, the occurrence of colitis was observed over a longer period (>3 months), which caused changes in the dosing regimen. The protocol is amended to include a modified dosing schedule, wherein based on PK simulations, weekly dosing (see discussion) is used after daily dosing. Patients at enrollment or after this protocol amendment received Compound 1C 20 mg QD for the first 9 weeks, followed by Compound 1C 20 mg once weekly (QW); patients who had received Compound 1C 20 mg QD for ≥9 weeks were switched to the QW schedule. Per the protocol amendment, ongoing and newly enrolled patients were required to receive prophylaxis against Pneumocystis jiroveci pneumonia (PJP).

[0390] Table 2A. Definition of dose-limiting toxicities

[0391]

[0392]

[0393] *Istatinib can cause a transient decrease in white blood cells due to margination; therefore, the DLT rule requires that neutropenia persist after 2 to 3 days of maintenance on itatinib. When the subject's clinical status permits, the investigator is prompted to wait 24 hours before starting growth factors to determine whether white blood cell margination contributes to the extent of neutropenia. AE: adverse event; ANC: absolute neutrophil count; DLT: dose-limiting toxicity.

[0394] For the Compound 1C + itatinib combination, a 3+3 design was used to perform dose escalation of Compound 1C, starting with a Compound 1C dose that was approximately 25% less than the recommended dose determined for Compound 1C monotherapy. The criteria for itatinib dose interruption are summarized in Table 2A. For each cohort, itatinib was co-administered at a dose of 300 mg QD.

[0395] For the Compound 1C + R-ICE combination, dose escalation of Compound 1C was performed using a 3+3 design with a starting dose approximately 25% lower than the recommended dose established for Compound 1C. The R-ICE chemotherapy component was administered according to institutional practice or per protocol.

[0396] Evaluate

[0397] Patients were evaluated on Days 1, 8, and 15 of Cycle 1 and on Day 1 (± 3) of each subsequent treatment cycle. The schedule of pivotal study assessments is shown in Table 2B.

[0398] Table 2B. Study Assessment

[0399]

[0400]

[0401] * Every 9 weeks (3 cycles) or as often as consistent with standard of care; performed only when measurable disease is present. May be performed according to standard of care; only for lymphoma. To confirm CR. § Not collected from patients receiving Compound 1C + R-ICE. || Cycle 2 only; collected from patients with B-cell malignancies in the Compound 1C monotherapy expansion cohort (Cohort A). Through cycle 6 for Compound 1C monotherapy and Compound 1C + itatinib combination; and in cycles 3, 6, 9, and 12 for Compound 1C + R-ICE combination. # Archival tissue is acceptable; for patients with CLL, peripheral blood is acceptable.** Optional on-treatment or EOT biopsy (or peripheral blood for patients with CLL). AE: adverse event; CLL: chronic lymphocytic leukemia; CR: complete response; CT: computed tomography; ECG: electrocardiogram; ECOGPS: Eastern Cooperative Oncology Group performance status; EOT: end of treatment; FDG-PET: fluorodeoxyglucose (FDG)-positron emission tomography; MRI: magnetic resonance imaging; PD: pharmacodynamics; PK: pharmacokinetics; R-ICE: rituximab plus ifosfamide, carboplatin, and etoposide.

[0402] The primary endpoint was the safety and tolerability of Compound 1C monotherapy, Compound 1C + itatinib, or Compound 1C + R-ICE, as assessed by a summary of treatment-emergent adverse events (TEAEs), clinical laboratory evaluations, physical examination results, and 12-lead electrocardiograms. The severity of TEAEs was assessed using Common Terminology Criteria for Adverse Events, version 4.03.

[0403] Secondary endpoints were efficacy and PK. Efficacy was measured by best overall response rate (ORR), defined as the proportion of patients achieving a partial response (PR) or complete response (CR). Responses were assessed every 9 weeks (per investigator assessment) based on the Lugano classification of lymphoma response criteria for HL and NHL (Cheson BD et al., Journal of Clinical Oncology, 2014; 32(27):3059-3067), the International Working Group criteria for chronic lymphocytic leukemia (CLL) (Hallek M. et al., Blood, 2008; 111(12):5446-5456; Cheson BD et al., J Clin Oncol, 2012; 30(23):2820-2822), and the VIth International Workshop on Waldenstrom's Macroglobulinemia Response Assessment for Waldenstrom's Macroglobulinemia (WM) (Owen RG et al., Br J Haematol, 2013; 160(2):171-176). If the patient's response to treatment was assessed by both PET and CT / MRI, the PET results were used for the response endpoint; if PET results were not available for assessment, CT / MRI results were used for assessment; if it was the only form of assessment available for the patient, CT / MRI was used for the response endpoint.

[0404] Exploratory endpoints included: duration of response (DOR, duration from first response to death or disease progression, whichever occurred first); progression-free survival (PFS, duration from first study dose to death or disease progression, whichever occurred first); population PK; and pharmacodynamic (PD) relationship with PI3Kδ inhibition (as determined by an ex vivo phosphorylated AKT [pAKT] assay).

[0405] Statistical methods

[0406] Safety, tolerability and efficacy analyses were performed on the safety / intent-to-treat population, including all recruited patients receiving ≥1 dose of any of Compound 1C, itatinib or R-ICE components. PK and PD analyses included all patients with available PK / PD data in the safety / intent-to-treat population. All statistical analyses were exploratory in nature and were summarized using descriptive statistics. ORR was estimated using a 95% confidence interval (CI) calculated using an exact method based on a binomial distribution. Kaplan-Meier estimates of median DOR and PFS were presented using their corresponding 95% CI.

[0407] Additional patient exclusion criteria

[0408] Additional patient exclusion criteria included radiation therapy within 4 weeks, investigational drug within 28 days (or 5 half-lives, whichever is longer), approved anticancer drug within 21 (42 for nitrosoureas) days (or 5 half-lives, whichever is longer) (except for steroids, ≤10 mg prednisone daily), unresolved toxicity grade ≥2, current or recent history of clinically significant infection, total bilirubin ≥1.2× upper limit of normal (ULN), alkaline phosphatase ≥2.5×ULN, aspartate aminotransferase (AST) or alanine aminotransferase ≥2.0×ULN, or creatinine clearance <50 mL / min based on the Cockroft-Gault formula.

[0409] R-ICE dosing schedule according to protocol

[0410] According to the protocol, R-ICE was administered in three 21-day cycles according to the following schedule: rituximab 375 mg / m on days 1 and 2 of cycle 1 and day 1 of cycles 2 and 3 2 On day 3 of each cycle, ifosfamide 5000 mg / m was given by continuous intravenous (IV) infusion over 24 h. 2 ; Carboplatin (AUC = 5 mg / mL; maximum dose 800 mg) by IV infusion on day 3; Etoposide 100 mg / m by IV on days 3 to 5 2 .

[0411] Example F. PI3Kδ inhibitor monotherapy

[0412] This analysis included data from patients enrolled in the Compound 1C monotherapy dose escalation (5-45 mg QD) and expansion (20 mg QD and 30 mg QD) cohorts.

[0413] Dose escalation and cohort expansion

[0414] The number of patients per dose level during Compound 1C monotherapy dose escalation and expansion is shown in Table 2C. No DLTs were identified, the MTD was not reached, and the 20 mg QD and 30 mg QD doses were expanded.

[0415] Table 2C. Monotherapy Dose Escalation and Cohort Expansion

[0416]

[0417] AE = adverse event; DLT = dose-limiting toxicity; MTD = maximum tolerated dose.

[0418] Pharmacokinetics and pharmacodynamics of compound 1C

[0419] All doses tested remained above the IC for target inhibition during the dosing interval. 90 ,like Figure 1A As shown; C generated by 20 mg QD dose of compound 1C 最大 and C 波谷 IC of pAKT inhibition 90 PK simulations were performed to support the selection of a 20 mg QW dosing schedule, estimating that serum Compound 1C levels produced by this schedule would exceed the IC for target inhibition for ∼36 hours. 90 and will continue for approximately half of the dosing interval with minimal to no inhibition. After single multiple dose administration of Compound 1C, Compound 1C rapidly reaches peak plasma concentrations with a median t 最大 0.5 to 1 hour. Subsequently, the plasma concentration of compound 1C decreased in a monophasic manner, with an average terminal-phase disposition of t 1 / 2 At steady state, dose proportionality appears to be observed between 5 and 45 mg QD, as Figure 1B shown.

[0420] Food effects were analyzed in 12 patients. Although co-administration of Compound 1C with a high-fat meal increased the 最大This was a 42% decrease, but it had a modest effect on the area under the curve (AUC; 10% decrease).The geometric mean ratio of fed to fasted AUC was 0.90 (90% CI: 0.68-1.18).

[0421] Near-maximal PI3Kδ inhibition was observed at all Compound 1C doses up to 6 hours after dosing, as measured by the effect on pAKT levels in whole blood, as Figure 4 shown.

[0422] Safety and Tolerability

[0423] TEAEs occurring in >10% of patients in all dose cohorts combined and in the 20 mg QD cohort are shown in Table 3.

[0424] Table 3. Non-hematologic TEAEs occurring in ≥15% of patients receiving Compound 1C monotherapy (safety population)

[0425]

[0426] *Based on starting dose. Includes patients receiving QD and QW dosing. The following preferences are included: diarrhea, colitis, enterocolitis, gastroenteritis, microscopic colitis, and cytomegalovirus colitis. Includes the following preferred terms: exfoliative dermatitis, rash, erythematous rash, macular rash, maculopapular rash, pruritic rash, exfoliative rash, generalized rash, epidemic rash, and pustular rash. MedDRA: Medical Dictionary for Regulatory Activities; TEAE: treatment-emergent adverse event.

[0427] The most common (≥30%) non-hematologic TEAEs of any grade in all patients were diarrhea / colitis (36%), nausea (36%), fatigue (31%), and rash (31%). One patient (1%) experienced grade 3 parenchymal inflammation and grade 3 cytomegalovirus colitis, respectively, and 7 patients (10%) experienced pneumonia (grade 3, n=3 [4%]). Laboratory abnormalities that occurred during treatment in ≥30% of patients are shown in Table 4. New onset / exacerbation of any grade and grade 3 / 4 neutropenia occurred in 32 patients (44%) and 14 patients (19%), respectively (Table 4). SAEs experienced by >2 patients were diarrhea / colitis (n=9 [13%]), pyrexia (n=4 [6%]), hypotension (n=3 [4%]), and sepsis (n=3 [4%]). Infections and infestations of any grade SAE (system organ class) occurred in 11 patients (15%), including pneumonia in 2 patients (3%).

[0428] Table 4. New or Worsening Hematologic Laboratory Abnormalities Emerging in ≥30% of Patients Receiving Compound 1C Monotherapy (Safety Population)*

[0429]

[0430] *Based on reported laboratory values. NA: Not Applicable

[0431] Of the TEAEs of interest, the median (range) time to onset of Grade 3 / 4 diarrhea / colitis was 5.7 (1.6-14.9) months and the median (range) time to onset of Grade 3 / 4 rash was 2.9 (1.5-9.3) months. AST and ALT elevations occurred in 21 (29%) and 20 (28%) patients, respectively; all events were Grade 1, with the exception of 4 events in 2 patients (1 patient had 1 event each of Grade 3 AST and Grade 2 ALT elevations, both occurring >30 days after the last dose of Compound 1C; 1 patient had 1 event each of Grade 3 AST and ALT elevations, considered secondary to sepsis). Bilirubin elevations occurred in 8 (11%) patients; all events were ≤ Grade 2, with the exception of 1 Grade 3 event that occurred >30 days after the date of the last dose. Changes in AST and ALT levels over time were reported in 21 (29%) and 20 (28%) patients, respectively; all events were Grade 1, with the exception of 4 events in 2 patients (1 patient had 1 event each of Grade 3 AST and Grade 2 ALT elevations, both occurring >30 days after the last dose of Compound 1C; 1 patient had 1 event each of Grade 3 AST and ALT elevations, considered secondary to sepsis). Figure 5A-5B There were no TEAEs of PJP or intestinal perforation. Five patients (7%) experienced hypertension (all grade 1 / 2); seven (10%) experienced hyperglycemia (all grade 1 / 2, except one grade 3).

[0432] TEAEs of any grade led to Compound 1C dosing interruptions in 30 patients (42%), dose reductions in 4 patients (6%), and treatment discontinuation in 14 patients (19%). The most common non-hematologic TEAEs leading to Compound 1C dosing interruptions were diarrhea / colitis (11 / 72 [15%]) and pyrexia (4 / 72 [6%]). TEAEs leading to death occurred in 2 patients (respiratory failure, and respiratory failure and sepsis); neither was considered related to Compound 1C.

[0433] Long-term tolerability and administration

[0434] Prior to implementation of the QW dosing schedule, 9 / 31 patients (29%) with DLBCL, FL, MCL, and MZL receiving compound 1C monotherapy discontinued due to TEAEs, the most common (n≥2) being diarrhea / colitis (n=3 [10%]) or rash (n=2 [6%]); however, no patient discontinued due to treatment-related AEs during the first 9 weeks of treatment. As of the data cutoff date, 26 of 72 enrolled patients (36%) had received QW dosing of compound 1C (after receiving QD dosing for at least 9 weeks) for a median (range) of 2.5 (1.2-9.6) months and a total of 105 patient months; 46 patients (64%) received QD dosing only; no patient was still in the first 9 weeks of QD dosing. 4 of 26 patients started the QW schedule at a dose of <20 mg due to a previous dose reduction (1 at 5 mg and 3 at 10 mg). Of the 26 patients who received QW dosing, 8 (31%) had dose interruptions (TEAEs leading to dose interruption in >1 patient were diarrhea and neutropenia [n=2 each]), 1 (4%) had dose reduction (due to pustular rash), and no patient discontinued treatment due to TEAEs. Four patients (15%) who received QW dosing experienced a total of 6 SAEs during the QW dosing period (abdominal pain, diarrhea, fever, bronchitis, sepsis, and dehydration, n=1 each). No grade 4 non-hematological TEAEs were reported during QW dosing. Two patients experienced grade 3 diarrhea and rash (n=1 [4%] each); both events occurred shortly after the switch from QD dosing. No new / worsening grade 4 neutropenia was reported, and one patient reported new / worsening grade 4 thrombocytopenia during QW dosing.

[0435] effect

[0436] The ORR in evaluable patients varied by disease type, as shown in Table 5. Among patients with evaluable lesion size at baseline and postbaseline, deep responses (>75% reduction in target lesion size from baseline) occurred in 4 / 7 patients (57%) with marginal zone lymphoma (MZL), 7 / 14 patients (50%) with follicular lymphoma (FL), 3 / 8 patients (38%) with mantle cell lymphoma (MCL), and 2 / 20 patients (10%) with diffuse large B-cell lymphoma (DLBCL) ( Figure 2 In patients with these tumor types, the majority (93%) of responses occurred at the first assessment (~9 weeks), as Figure 6 Durable responses (>6 months) were observed in patients receiving QW dosing and several patients remained on the QW schedule for at least 8 months, as shown in Figure 3As shown, including 2 patients who received 10 mg QW. The median duration of response was 13.5 months (95% CI: 8.3-18.8) for DLBCL, 4.4 months (95% CI: 2.1-NE) for MZL, and was not reached in FL or MCL. Figure 7 Representative positron emission tomography (PET) images of patients who achieved complete responses to compound 1C monotherapy are shown. Patients <65 years old, with mantle cell lymphoma without bone marrow involvement (target lesion, 98 mm × 58 mm) and who had received 1 prior treatment (R-HyperCVAD). After treatment with compound 1C (30 mg, once daily), the patient achieved complete response at week 9.

[0437] Table 5. Best Overall Response in Patients Receiving Compound 1C Monotherapy*

[0438]

[0439] * Assessed by: Lugano classification, IWGC criteria for CLL, and VIth International Workshop on WM, and by CT or PET, depending on disease subtype. The 95% CI was calculated based on the exact method for the binomial distribution. Two patients had unknown DLBCL subtype. These included extranodal MZL of MALT type (n=2), nodular MZL (n=4), splenic MZL (n=2), and unknown MZL subtype (n=1). ∥∥ Four patients with MCL and three patients with CLL had received ibrutinib prior to the study, with a best overall response of CR or PR achieved in two patients with MCL and one patient with CLL. ** Classical HL (n=9) and nodular lymphocyte-predominant HL (n=1) were included. ABC: activated B-cell-like; CI: confidence interval; CLL: chronic lymphocytic leukemia; CMR: complete metabolic response; CR: complete response; CT: computed tomography; DLBCL: diffuse large B-cell lymphoma; FL: follicular lymphoma; HL: Hodgkin lymphoma; GCB: germinal center B-cell-like; CLL IWGC: International Working Group on CLL; MALT: mucosa-associated lymphoid tissue; MCL: mantle cell lymphoma; MZL: marginal zone lymphoma; ORR: overall response rate; PD: progressive disease; PET: positron emission tomography; PMD: progressive metabolic disease; PMR: partial metabolic response; PR: partial response; SD: stable disease; WM: Waldenstrom's macroglobulinemia.

[0440] Example G. PI3Kδ inhibitor combination therapy

[0441] Compound 1C in combination with itatinib

[0442] 11 patients were recruited and treated with compound 1C + itatinib. The median (range) duration of treatment with compound 1C and itatinib was 2.1 (1.0-10.3) months. At the time of data cutoff, treatment was performed in 2 patients (18%); 8 patients (73%) discontinued treatment due to disease progression, and 1 (9%) discontinued treatment due to TEAE.

[0443] Of the 11 patients treated with Compound 1C + itatinib, 8 received Compound 1C 20 mg QD and 3 received Compound 1C 30 mg QD; all patients received itatinib 300 mg QD. No DLTs were observed. Nine patients (82%) experienced TEAEs, 5 (45%) had grade 3 / 4 TEAEs, and 2 (18%) had SAEs. The most common TEAEs of any grade (n≥2) were nausea, fatigue (n=5 each), cough, hypertension, vomiting (n=3 each), decreased appetite, hyperglycemia, increased tearing, oral herpes, and tachycardia (n=2 each). Five patients experienced grade 3 / 4 TEAEs: grade 3 spinal cord compression, grade 3 intractable pain, and grade 3 alkaline phosphatase increase (n=1); grade 3 worsening hyperlipidemia (n=1); grade 3 hypertension (n=1); grade 3 diarrhea and grade 3 dehydration (n=1); and grade 3 hypertension and grade 4 hypercalcemia (n=1). One patient had a new / worsening grade 4 blood laboratory abnormality (thrombocytopenia). One SAE each was reported: grade 3 dehydration and grade 4 hypercalcemia. Two patients (18%) had TEAEs that led to dose interruptions of compound 1C and itatinib, and 1 patient (9%) had a TEAE that led to discontinuation of compound 1C and itatinib; no patient had a TEAE that led to dose reduction. No TEAE resulted in death.

[0444] Among evaluable patients, patients with CLL (1 / 1) and FL (1 / 1) achieved a best overall response of CR / complete metabolic response; patients with MCL (1 / 1) and classic HL (1 / 2) achieved a best overall response of partial metabolic response. All 6 patients with DLBCL and 1 of 2 patients with classic HL had a best overall response of PD / progressive metabolic disease.

[0445] Compound 1C in combination with R-ICE

[0446] Five patients with DLBCL were treated with compound 1C + R-ICE. The median (range) duration of compound 1C treatment was 2.3 (1.5-3.7) months; 4 patients each received 3 cycles of R-ICE, and 1 patient received 2 cycles of R-ICE. As of the data cutoff date, all patients discontinued treatment due to physician decision (n = 2), progressive disease, adverse events, and others (n = 1 each). Of the 5 patients, 4 received compound 1C 15mg QD and 1 received 20mg QD. No DLT was observed. The most common non-hematological TEAEs of any grade (≥2 patients) were dizziness (n = 3), alopecia, constipation, excess body fluid, headache, hypokalemia, and night sweats (n = 2 each). Three patients reported new / worsened grade 4 thrombocytopenia and two patients reported new / worsened grade 4 neutropenia. Two patients (40%) experienced SAEs, 1 patient experienced Grade 3 acute encephalopathy (related to ifosfamide), and 1 experienced Grade 2 atrial flutter and atrial fibrillation and Grade 3 dyspnea, neutropenia, and febrile neutropenia (all related to Compound 1C and R-ICE). Three patients (60%) had TEAEs that led to dose interruptions of Compound 1C and R-ICE, 1 patient (20%) had a TEAE that led to dose interruption of Compound 1C only, and 1 patient (20%) had a TEAE that led to discontinuation of Compound 1C and R-ICE; no patient had a TEAE that led to dose reduction. No TEAEs resulted in death.

[0447] Of the 5 patients treated with Compound 1C + R-ICE, 3 patients achieved a complete metabolic response (1 of the 3 patients indicated continued SCT) and 2 patients had stable disease as best response.

[0448] Example H. Results of PI3Kδ Monotherapy and Combination Therapy

[0449] Compound 1C exhibits a differential safety profile relative to first-generation PI3K inhibitors (e.g., idelalisib and duvelisib) (Coutré S.E. et al., Leukemia & Lymphoma. 2015; 56(10):2779-2786; Flinn IW et al., Blood, 2018; 131(8):877-887), most notably indicated by the near absence of grade ≥2 transaminase elevations. Compound 1C is comparable to copanlisib, a pan-PI3K inhibitor approved in the United States as monotherapy for the treatment of relapsed FL (ALIQOPA TM(Kupanisib), for injection, intravenous use, US Whippany, NJ: Bayer HealthCare Pharmaceuticals Inc.; 2017), there was no clinically significant hypertension or hyperglycemia, presumably due to the high selectivity of compound 1C for the delta isoform of PI3K. Although no PJP was observed, the protocol amendment made a mandatory PJP prevention component in response to PJP infections reported in a different Phase 1 study by studying compound 1C in combination with pembrolizumab.

[0450] Diarrhea / colitis and rash (common toxicity of PI3K inhibitors) were the 2 most frequent toxicities observed in this study; however, most diarrheal events were grade 1 or 2 and were manageable. Most grade 3 / 4 diarrhea / colitis and rash occurred after the first disease assessment; overall, diarrhea / colitis or rash of any grade led to treatment discontinuation in 9 / 72 (13%) patients, including 3 patients with diarrhea / colitis who were positive after re-challenge with the same dose. Grade 4 neutropenia (6%) was observed only during QD dosing (first occurring after 21 days of treatment). The safety profile associated with the 20 mg QD dose was generally consistent with that observed for all combined doses and with that observed for each individual dose level (data not shown), but relatively few patients were administered at levels below 20 mg QD and above 30 mg QD.

[0451] Long-term administration of compound 1C causes delayed TEAE, resulting in treatment interruption. In order to improve the long-term tolerability of compound 1C while maintaining a high response rate, a modified dosing regimen (20 mg QD for 9 weeks, followed by 20 mg QW) was implemented. The daily dosing period was maintained to retain the possibility of rapid response (at the time of implementation, all 11 patients with NHL who were administered 20 mg QD had achieved the target response, 10 of whom appeared at the time of the first disease assessment). The intermittent portion of the dosing regimen (implemented to reduce the incidence of delayed TEAE) was based on comparative PK / PD simulations using Kupanisi (Patnaik A. et al., Annals of Oncology, 2016; 27 (10): 1928-1940). Without being bound by theory, it is expected that the 20 mg QW regimen provides similar pAKT inhibition as expected for the QW administration of Kupanisi. Encouragingly, based on data accumulated during the 105 patient-month period, none of the 26 patients who received QW administration discontinued the study treatment due to TEAE. This observation, combined with the absence of colitis, grade 4 non-hematological TEAEs and neutropenia, and low incidence of grade 3 diarrhea and rash (n=1 each), suggests that the QW schedule is better tolerated than the continuous QD schedule using the doses evaluated herein.

[0452] Compound 1C monotherapy achieved high rates of rapid, deep, and durable objective responses at doses ranging from 10 mg QD to 45 mg QD in relapsed or refractory B-cell NHL. ORRs in all subtypes ranged from 30% to 78% (DLBCL: 30%; MCL: 67%; FL: 71%; MZL: 78%) and deep responses occurred in >30% of patients. Interestingly, objective responses were observed in 2 of 4 patients with MCL who had previously been treated with ibrutinib; 1 of these responses was ongoing at approximately 6 months at the time of data cutoff. Among patients with DLBCL, substantially more patients with the GCB subtype (n=19) were recruited relative to those with the ABC subtype (n=2). These small, disproportionate numbers are insufficient to determine whether compound 1C is more active in one subtype than in others. Durable responses were observed in all 4 of these NHL subtypes, including ~17 months in FL, ~10 months in MZL, ≥14 months in MCL, and ~19 months in DLBCL, all of which included QW dosing and 2 of which (FL and MZL) had ongoing responses at the time of data cutoff. Taken together, these data suggest that QW dosing of Compound 1C can achieve durable responses in patients with B-cell NHL.

[0453] PK and safety data from the combination study showed that compound 1C can be safely combined with itatinib at a 300 mg QD dose. Similarly, the combination of compound 1C with R-ICE also appears to be tolerated. Additional studies are needed to evaluate the safety of compound 1C in combination with R-ICE and the potential for improved efficacy for each combination.

[0454] Compound 1C exhibits antitumor activity with rapid and deep responses, high response rates, and a differential toxicity profile in patients with relapsed or refractory B-cell malignancies. The modified dosing regimen described herein appears to improve the long-term tolerability of Compound 1C and is able to achieve durable responses in several patients. The efficacy, safety, PK, and PD data presented here support further investigation of Compound 1C monotherapy in selected NHL subtypes (DLBCL [NCT02998476], FL [NCT03126019], MZL [NCT03144674], and MCL [NCT03235544]) in an ongoing Phase 2 study. Phase 1 studies are also ongoing, exploring several combination strategies in patients with NHL (NCT03039114; NCT03424122).

[0455] Example I. Comparison of maintenance dosing

[0456] In this study, subjects were administered 20 mg of Compound 1C once daily for 8 weeks in combination with a JAK1 inhibitor. After Week 8, subjects were assigned to 1 of 2 maintenance dose groups based on block randomization. In one group, subjects were administered 5 mg of Compound 1C once daily. In the second group, subjects were administered 20 mg of Compound 1C once a week.

[0457] The safety and tolerability of Compound 1C were assessed by a summary of treatment-emergent adverse events (TEAEs), clinical laboratory assessments, physical examination results, and 12-lead electrocardiograms. The severity of TEAEs was assessed using Common Terminology Criteria for Adverse Events version 4.03. Secondary endpoints were efficacy and PK, as described in Example E.

[0458] Without being bound by theory, it is believed that a 5 mg once daily maintenance dose of Compound 1C provides similar safety and tolerability as a 20 mg once weekly maintenance dose of Compound 1C.

[0459] Example J. PI3Kδ Combination Therapy with JAK1 / 2 Inhibitors in the Treatment of Myelofibrosis

[0460] Compound 1C in combination with ruxolitinib was evaluated in patients with primary or secondary myelofibrosis (MF) (e.g., PMF, PPV-MF, or PET-MF) who showed a suboptimal response to ruxolitinib monotherapy. The study was designed in three parts, as follows:

[0461] Part 1, Safety Run-in : 3+3 design; Ruxolitinib + Compound 1C, administered at a dose of 10 mg or 20 mg QD for 8 weeks, followed by 10 mg or 20 mg QW.

[0462] Part 2, randomization period (1:1) : Group 1: Ruxolitinib + Compound 1C, administered at a dose of 10 mg QD for 8 weeks, followed by 10 mg QW. Group 2: Ruxolitinib + Compound 1C, administered at a dose of 20 mg QD for 8 weeks, followed by 20 mg QW.

[0463] Part 3 : Alternative dosing regimens, including continuous daily dosing.

[0464] A suboptimal response to ruxolitinib monotherapy was defined as follows:

[0465] Patients have been treated with ruxolitinib for ≥6 months, on a stable dose, for ≥8 weeks immediately prior to enrollment (acceptable dose of prior ruxolitinib: 5 mg–25 mg BID); and

[0466] A spleen palpable >10 cm below the left costal margin during physical examination at Screening; or

[0467] At the screening visit, a palpable spleen 5-10 cm below the left subcostal margin and active symptoms of myelofibrosis during physical examination, each defined as 1 symptom score ≥5; or 2 symptom scores ≥3, using the following screening symptom form: 10-point scale for each of 7 symptoms. Symptoms include: night sweats, itching, abdominal discomfort, left subcostal pain, early satiety, bone / muscle pain, and immobility.

[0468] The dosing regimen and preliminary results of the Compound 1C + Ruxolitinib study are shown in Appendix A, the disclosure of which is incorporated herein by reference in its entirety. As shown in Appendix A, add-on therapy with Compound 1C appears to provide additional clinical benefit in patients with MF who have a suboptimal response to Ruxolitinib monotherapy. Spleen volume reduction was observed in 56% of patients at Week 12, with symptom improvement occurring as early as 4 weeks; the median symptom improvement at Week 24 was 36%. The combination of Compound 1C and Ruxolitinib was well tolerated in patients, with no dose-limiting toxicity (DLT) in Part 1 and with the expected Grade 3 / 4 side effect (AE) profile. Of the 17 patients treated for >6 months, only 2 (7%) discontinued due to AEs. AEs common to PI3Kδ inhibitors (such as liver, rash, and colitis) are rare.

[0469] In addition to those described herein, various modifications of the present invention will be apparent to those skilled in the art from the above description. Such modifications are also intended to fall within the scope of the appended claims. Each reference cited in this application, including all patents, patent applications and publications, is incorporated herein by reference in its entirety.

[0470] In summary, the present invention includes but is not limited to the following items:

[0471] 1. A method for treating a disease in a patient, wherein the disease is associated with abnormal expression or activity of PI3Kδ kinase, the method comprising:

[0472] i) administering to the patient a first dose of about 3 mg / day to about 50 mg / day of a PI3Kδ inhibitor for a first period of about 2 weeks to about 12 weeks; and

[0473] ii) administering to the patient a second dose of the PI3Kδ inhibitor at the end of the first period, the second dose being less than the first dose and being:

[0474] (a) about 2.5 mg / day or less; or

[0475] (b) about 50 mg / week or less;

[0476] And wherein the second dose is administered during a second period occurring after the first period.

[0477] 2. The method of claim 1, wherein the first dose is about 20 mg / day.

[0478] 3. The method of claim 1 or 2, wherein the first period is from about 8 weeks to about 12 weeks.

[0479] 4. A method as described in claim 1 or 2, wherein the first period is about 8 weeks.

[0480] 5. A method as described in any one of items 1 to 4, wherein the first dose is reduced during the first time period.

[0481] 6. The method of any one of items 1 to 5, wherein the second dose is about 2.5 mg / day or less.

[0482] 7. The method of any one of items 1 to 5, wherein the second dose is about 2.5 mg / day.

[0483] 8. The method of any one of items 1 to 5, wherein the second dose is about 50 mg / week or less.

[0484] 9. The method of any one of items 1 to 5, wherein the second dose is about 20 mg / week.

[0485] 10. The method of any one of items 1 to 9, wherein the second dose is reduced during the second period.

[0486] 11. A method as described in any one of items 1 to 10, wherein the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs).

[0487] 12. A method as described in any one of items 1 to 11, wherein the patient has been identified as exhibiting one or more symptoms associated with one or more treatment-emergent adverse events (TEAEs) during the first period.

[0488] 13. A method as described in any one of items 1 to 12, wherein the first period ends when the patient has been identified as exhibiting one or more symptoms associated with one or more adverse events (TEAEs) occurring during treatment.

[0489] 14. A method as described in any one of items 1 to 13, wherein the adverse events occurring during the one or more treatments include one or more of the following: diarrhea / colitis, nausea, fatigue, rash, neutropenia, fever, hypotension, sepsis, respiratory failure, lung parenchymal inflammation, pneumonia, hypertension, hyperglycemia, abdominal pain, bronchitis, dehydration, thrombocytopenia, cough, vomiting, loss of appetite, increased tearing, oral herpes, tachycardia, spinal cord compression, intractable pain, increased alkaline phosphatase, increased transaminases, hyperlipidemia, hypercalcemia, dizziness, alopecia, constipation, excessive body fluid, headache, hypokalemia, night sweats, encephalopathy, atrial flutter, atrial fibrillation and dyspnea.

[0490] 15. A method as described in any one of items 1 to 13, wherein the adverse events occurring during the one or more treatments include one or more of the following: diarrhea / colitis, nausea, fatigue, rash, cough, vomiting, dizziness, fever, hypokalemia, abdominal pain, constipation, loss of appetite, night sweats, itching, back pain, chills, leukopenia, neutropenia, lymphopenia, thrombocytopenia and anemia.

[0491] 16. A method as described in any one of items 1 to 13, wherein the adverse events occurring during the one or more treatment periods include one or more of diarrhea / colitis and rash.

[0492] 17. The method of any one of items 14 to 16, wherein the diarrhea / colitis comprises one or more of the following: diarrhea, colitis, enterocolitis, gastroenteritis, microscopic colitis, and cytomegalovirus colitis.

[0493] 18. A method as described in any one of items 14 to 17, wherein the rash comprises one or more of the following: exfoliative dermatitis, rash, erythematous rash, macular rash, maculopapular rash, pruritic rash, exfoliative rash, generalized rash, epidemic rash, and pustular rash.

[0494] 19. A method as described in any one of items 14 to 18, wherein the neutropenia comprises febrile neutropenia.

[0495] 20. The method of any one of items 14 to 19, wherein the elevated transaminase comprises elevated alanine aminotransferase (ALT), elevated aspartate aminotransferase (AST), or a combination thereof.

[0496] 21. The method of any one of items 1 to 20, wherein the PI3Kδ inhibitor is selected from:

[0497] 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one; and

[0498] 5-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}-1,3-oxazolidin-2-one;

[0499] or a pharmaceutically acceptable salt thereof.

[0500] 22. A method as described in any one of items 1 to 20, wherein the PI3Kδ inhibitor is 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one or a pharmaceutically acceptable salt thereof.

[0501] 23. A method as described in claim 22, wherein the PI3Kδ inhibitor is (S)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one or a pharmaceutically acceptable salt thereof.

[0502] 24. A method as described in claim 22, wherein the PI3Kδ inhibitor is (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one or a pharmaceutically acceptable salt thereof.

[0503] 25. A method as described in claim 22, wherein the PI3Kδ inhibitor is (S)-4-(3-((R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one or a pharmaceutically acceptable salt thereof.

[0504] 26. A method as described in claim 22, wherein the PI3Kδ inhibitor is (R)-4-(3-((R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one or a pharmaceutically acceptable salt thereof.

[0505] 27. A method as described in claim 22, wherein the PI3Kδ inhibitor is a pharmaceutically acceptable salt of 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one.

[0506] 28. A method as described in claim 27, wherein the PI3Kδ inhibitor is (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride.

[0507] 29. A method as described in claim 27, wherein the salt is a salt in which the stoichiometric ratio of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one to hydrochloric acid is 1:1.

[0508] 30. A method as described in claim 27, wherein the salt is crystalline.

[0509] 31. A method as described in any one of items 1 to 20, wherein the PI3Kδ inhibitor is 5-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}-1,3-oxazolidin-2-one or a pharmaceutically acceptable salt thereof.

[0510] 32. A method as described in claim 31, wherein the PI3Kδ inhibitor is (R)-5-{3-[(R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}-1,3-oxazolidin-2-one or a pharmaceutically acceptable salt thereof.

[0511] 33. A method as described in claim 31, wherein the PI3Kδ inhibitor is (R)-5-{3-[(S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}-1,3-oxazolidin-2-one or a pharmaceutically acceptable salt thereof.

[0512] 34. A method as described in claim 31, wherein the PI3Kδ inhibitor is (S)-5-{3-[(S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}-1,3-oxazolidin-2-one or a pharmaceutically acceptable salt thereof.

[0513] 35. A method as described in claim 31, wherein the PI3Kδ inhibitor is (S)-5-{3-[(R)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}-1,3-oxazolidin-2-one or a pharmaceutically acceptable salt thereof.

[0514] 36. A method as described in any one of items 1 to 35, wherein the disease is selected from chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Hodgkin lymphoma (HL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL) and Waldenstrom's macroglobulinemia (WM).

[0515] 37. The method of claim 36, wherein the marginal zone lymphoma (MZL) is selected from the group consisting of extranodal MZL, nodal MZL, splenic MZL, and unknown MZL subtype.

[0516] 38. The method of claim 36, wherein the Hodgkin's lymphoma (HL) is selected from classical Hodgkin's lymphoma (HL) and nodular lymphocyte-predominant HL.

[0517] 39. A method as described in claim 36, wherein the diffuse large B-cell lymphoma is selected from activated B-cell-like (ABC) diffuse large B-cell lymphoma (ABC-DLBCL) and germinal center B cell (GCB) diffuse large B-cell lymphoma (GCB-DLBCL).

[0518] 40. A method of treating a disease in a patient, wherein the disease is selected from the group consisting of chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Hodgkin lymphoma (HL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), and Waldenstrom's macroglobulinemia (WM), the method comprising:

[0519] i) administering to the patient a first dose of about 20 mg / day to about 50 mg / day of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride over a first period of about 8 weeks to about 9 weeks; and

[0520] ii) administering to the patient a second dose of the (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride salt during a second period occurring after the first period, the second dose being about 2.5 mg / day or less.

[0521] 41. A method of treating a disease in a patient, wherein the disease is selected from the group consisting of chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Hodgkin lymphoma (HL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), and Waldenstrom's macroglobulinemia (WM), the method comprising:

[0522] i) administering to the patient a first dose of about 20 mg / day to about 50 mg / day of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride over a first period of about 8 weeks to about 9 weeks; and

[0523] ii) administering to the patient a second dose of the (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride salt in a second period occurring after the first period, the second dose being about 20 mg / week to about 50 mg / week.

[0524] 42. A method of treating a disease in a patient, wherein the disease is selected from the group consisting of chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Hodgkin lymphoma (HL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), and Waldenstrom's macroglobulinemia (WM), the method comprising:

[0525] i) administering to the patient a first dose of about 20 mg / day of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride over a first period of about 8 weeks; and

[0526] ii) administering to the patient a second dose of the (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride salt in a second period occurring after the first period, the second dose being about 2.5 mg / day.

[0527] 43. A method of treating a disease in a patient, wherein the disease is selected from the group consisting of chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Hodgkin lymphoma (HL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), and Waldenstrom's macroglobulinemia (WM), the method comprising:

[0528] i) administering to the patient a first dose of about 20 mg / day of (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride over a first period of about 8 weeks; and

[0529] ii) administering to the patient a second dose of the (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride salt in a second period occurring after the first period, the second dose being about 20 mg / week.

[0530] 44. A method of treating a disease in a patient, wherein the disease is associated with abnormal expression or activity of PI3Kδ kinase, the method comprising:

[0531] i) administering to the patient a first dose of about 3 mg / day to about 50 mg / day of a PI3Kδ inhibitor for a first period of about 2 weeks to about 12 weeks; and

[0532] ii) administering to the patient a second dose of the PI3Kδ inhibitor at the end of the first period, the second dose being less than the first dose and between about 2.5 mg / day and about 7.5 mg / day; and

[0533] wherein the second dose is administered during a second period occurring after the first period.

[0534] 45. The method of claim 44, wherein the second dose is about 3.0 mg / day to about 7.0 mg / day.

[0535] 46. ​​The method of claim 44, wherein the second dose is about 4.0 mg / day to about 6.0 mg / day.

[0536] 47. The method of claim 44, wherein the second dose is about 5.0 mg / day.

[0537] 48. A method as described in any one of items 44 to 47, wherein the PI3Kδ inhibitor is (R)-4-(3-((S)-1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one hydrochloride.

[0538] 49. A method as described in any one of items 44 to 48, wherein the disease is selected from chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Hodgkin lymphoma (HL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL) and Waldenstrom's macroglobulinemia (WM).

[0539] 50. The method of claim 49, wherein the marginal zone lymphoma (MZL) is selected from the group consisting of extranodal MZL, nodal MZL, splenic MZL, and unknown MZL subtype.

[0540] 51. The method of claim 49, wherein the Hodgkin's lymphoma (HL) is selected from classical Hodgkin's lymphoma (HL) and nodular lymphocyte-predominant HL.

[0541] 52. A method as described in claim 49, wherein the diffuse large B-cell lymphoma is selected from activated B-cell-like (ABC) diffuse large B-cell lymphoma (ABC-DLBCL) and germinal center B cell (GCB) diffuse large B-cell lymphoma (GCB-DLBCL).

Claims

1. Use of a PI3Kδ inhibitor in the preparation of a medicament for treating a disease in a patient, wherein the disease is selected from chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Hodgkin lymphoma (HL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL) and Waldenstrom's macroglobulinemia (WM), wherein: i) the medicament comprises a first dose of the PI3Kδ inhibitor, the first dose is about 3 mg / day to about 50 mg / day, and is used for a first period of about 2 weeks to about 12 weeks; and ii) the medicament comprises a second dose of the PI3Kδ inhibitor, the second dose is used at the end of the first period, the second dose is less than the first dose and is: (a) about 2.5 mg / day or less; or (b) about 50 mg / week or less; and wherein the second dose is administered during a second period occurring after the first period; and The PI3Kδ inhibitor is 4-{3-[1-(4-amino-3-methyl-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl]-5-chloro-2-ethoxy-6-fluorophenyl}pyrrolidin-2-one or a pharmaceutically acceptable salt thereof.

2. The method of claim 1, wherein the first dose is about 20 mg / day.

3. The use of claim 1 or 2, wherein the first period is from about 8 weeks to about 12 weeks.

4. The use of claim 1 or 2, wherein the first period is about 8 weeks.

5. The use of any one of claims 1 to 4, wherein the first dose is reduced during the first period.

6. The use of any one of claims 1 to 5, wherein the second dose is about 2.5 mg / day or less.

7. The use of any one of claims 1 to 5, wherein the second dose is about 2.5 mg / day.

8. The use of any one of claims 1 to 5, wherein the second dose is about 50 mg / week or less.

9. The use of any one of claims 1 to 5, wherein the second dose is about 20 mg / week.

10. The use of any one of claims 1 to 9, wherein the second dose is reduced during the second period.

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