Treatment of diseases and disorders with IRAK4-modified compounds

Abstract

The present disclosure relates to methods of treating diseases and conditions with IRAK4-modified compounds, wherein the diseases and conditions are characterized by the presence of a related biomarker in a biological sample of a subject.

Description

[0001] Related Applications

[0002] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 340,692, filed on May 11, 2022, the contents of which are incorporated herein by reference in their entirety. Background Art

[0003] Interleukin-1 (IL-1) receptor-associated kinase 4 (IRAK4) is a serine / threonine kinase that plays an important role in signal transduction through Toll / IL-1 receptor (TIR). Various IRAK enzymes are key components in signal transduction pathways mediated by interleukin-1 receptor (IL-1R) and Toll-like receptors (TLR) (Janssens, S et al., Mol. Cell. 11, 2003, 293–302). There are four members in the mammalian IRAK family: IRAK1, IRAK2, IRAK3 and IRAK4. These proteins are characterized by a typical N-terminal death domain that mediates interaction with the MyD88 family adaptor protein and a centrally located kinase domain. IRAK proteins, as well as MyD88, have been shown to play a role in transducing signals other than those derived from the IL-1R receptor, including those triggered by the IL-18 receptor (Kanakaraj et al., J. Exp. Med. 189 (7): 1999, 1129-38) and by activation of the LPS receptor (Yang et al., J. Immunol. 163, 1999, 639-643). Of the four members of the mammalian IRAK family, IRAK4 is considered to be a "motherboard IRAK". Under overexpression conditions, all IRAKs can mediate activation of nuclear factor-κB (NF-κB) and stress-induced mitogen-activated protein kinase (MAPK)-signaling cascades. However, only IRAK-1 and IRAK4 have been shown to have active kinase activity. Although the kinase activity of IRAK-1 may be dispensable for its function in IL-1-induced NF-κB activation (Kanakaraj et al., J. Exp. Med. 187(12), 1998, 2073–2079) and (Xiaoxia Li et al., Mol. Cell. Biol. 19(7), 1999, 4643–4652), the signal transduction of IRAK4 requires its kinase activity (Li S et al., Proc. Natl. Acad. Sci. USA 99(8), 2002, 5567–5572) and (Lye, E et al., J. Biol. Chem. 279(39); 2004, 40653-8). Given the central role of IRAK4 in Toll-like / IL-1R signaling and immune protection, IRAK4 inhibitors have been considered as valuable therapeutic agents in inflammatory diseases, sepsis and autoimmune diseases (Wietek C et al., Mol. Interv. 2:2002, 212-215).

[0004] Mice lacking IRAK4 are viable and show complete abrogation of inflammatory cytokine production in response to IL-1, IL-18 or LPS (Suzuki et al., Nature, 416(6882), 2002, 750-756). Similarly, human patients lacking IRAK4 are severely immunocompromised and do not respond to these cytokines (Medvedev et al., J. Exp. Med., 198(4), 2003, 521-531 and Picard et al., Science 299(5615), 2003, 2076-2079). Knock-in mice containing inactive IRAK4 are completely resistant to lipopolysaccharide and CpG-induced shock (Kim TW et al., J Exp Med 204:2007, 1025-36) and (Kawagoe T et al., J Exp Med 204(5):2007, 1013-1024) and IRAK4 kinase activity was shown to be essential for cytokine production, activation of MAPKs and induction of NF-κB-regulated genes in response to TLR ligands (Koziczak-Holbro M et al., J Biol Chem; 282(18):2007; 13552-13560). Inactivation of IRAK4 kinase (IRAK4 KI) in mice results in resistance to EAE due to a reduction in inflammatory cells infiltrating into the CNS and reduced IL-17 production mediated by antigen-specific CD4+ T cells (Kirk A et al., The Journal of Immunology, 183(1), 2009, 568-577).

[0005] Non-Hodgkin lymphoma (NHL) is the most common hematological malignancy in adults, with an estimated 78,000 new cases and 20,000 deaths in the United States in 2020. The molecular pathology that drives NHL is diverse, but a common theme is the overactivity of the NF-κB signaling pathway. Specific molecular changes that drive this pathway have been identified as a subset of NHL. For example, diffuse large B-cell lymphoma (hereinafter also referred to as "DLBCL") is an aggressive lymphoma that can occur in lymph nodes or outside the lymphatic system, in the gastrointestinal tract, testicles, thyroid, skin, breast, bone or brain. DLBCL is a cancer of B cells, a type of white blood cell responsible for producing antibodies. It is the most common type of non-Hodgkin lymphoma in adults, with an incidence of 7-8 cases per 100,000 people per year. This cancer occurs primarily in older individuals, with a median age of diagnosis of approximately 70 years, but can also occur in children and young adults in a few cases. DLBCL is an aggressive tumor, and the first symptom of the disease is usually a rapidly growing mass. The five-year survival rate is only 58%. DLBCL has a subtype named according to its cell source, and includes germinal center B cell-like (GCB) and activated B cell-like (ABC). The difference is that it has a poor prognosis, and in some cases, requires special treatment methods.

[0006] Another example of NHL is Waldenstrom's macroglobulinemia (WM). WM is a non-Hodgkin lymphoma that affects two types of B cells, lymphoplasmacytoid cells and plasma cells. WM is characterized by high levels of circulating antibodies, immunoglobulin M (IgM), which are produced and secreted by cells involved in the disease. WM is a rare disease, with only about 1,500 cases per year in the United States. There is no single accepted treatment for WM, and clinical outcomes vary significantly due to gaps in knowledge about the molecular basis of the disease. Objective response rates are high (>80%), but complete response rates are low (0%-15%).

[0007] Other types of non-Hodgkin's lymphoma include mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), follicular lymphoma (FL), chronic lymphoblastic leukemia (CLL), small lymphocytic lymphoma (SLL), CNS lymphoma and testicular lymphoma. Non-Hodgkin's lymphoma can be caused by a variety of factors, such as infectious agents (Epstein-Barr virus, hepatitis C virus and human T-cell leukemia virus), radiation and chemotherapy treatments, and autoimmune diseases. As a group, non-Hodgkin's lymphoma affects 2.1% of the U.S. population during their lifetime. The percentage of people who survive more than five years after diagnosis is 71%.

[0008] In view of the above, there is a clear and unmet need for additional therapies for the treatment of cancer and other diseases associated with IRAK4. Summary of the invention

[0009] In certain aspects, the present disclosure provides a method of treating a disease or disorder in a subject, the method comprising:

[0010] obtaining one or more biological samples from a subject;

[0011] analyzing the biological sample for the presence of a first biomarker in nuclei of cells of the biological sample; and

[0012] administering to the subject an IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader when the presence of the first biomarker is detected in the nucleus of the cell of the biological sample;

[0013] The first biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

[0014] In some aspects, the present disclosure provides a method of treating a disease or disorder in a subject, the method comprising:

[0015] obtaining one or more biological samples from the subject;

[0016] measuring a sample expression level of a first biomarker in the biological sample;

[0017] comparing the sample expression level of the first biomarker to a reference expression level of the first biomarker;

[0018] administering to the subject an IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader when the sample expression level of the first biomarker in the sample is elevated compared to the reference expression level of the first biomarker; and

[0019] The first biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

[0020] In other aspects, the present disclosure provides methods of treating a disease or condition in a subject, the method comprising:

[0021] obtaining one or more biological samples from the subject;

[0022] measuring a sample expression level of a first biomarker in the biological sample;

[0023] comparing the sample expression level of the first biomarker to a reference expression level of the first biomarker;

[0024] analyzing the biological sample for the presence of the first biomarker in nuclei of cells of the biological sample; and

[0025] An IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader is administered to the subject only if:

[0026] detecting the presence of the first biomarker in the nucleus of the cell of the biological sample; and

[0027] the sample expression level of the first biomarker in the sample is increased compared to the reference expression level of the first biomarker;

[0028] The first biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 is a schematic diagram of a cancer cell showing the typical NF-κB signaling pathway and related processes and antibody complexes associated with the amount of IRAK4 expression in the membrane, cytoplasm, and nucleus.

[0030] Figure 2A Representative images of training samples including ICH-stained triple-positive bone marrow aspirate clot samples obtained from AML patients, showing the expression of IRAK4 in the samples. Blasts: 34%.

[0031] Figure 2B Representative images of training samples including ICH-stained triple-positive bone marrow aspirate clot samples obtained from AML patients, showing the expression of NF-κΒ p-p65 S536 in the samples. Blasts: 34%.

[0032] Figure 2C Representative images of training samples including ICH-stained triple-positive bone marrow aspirate clot samples obtained from AML patients, showing the expression of NF-κΒ p-p50 S337 in the samples. Blasts: 34%.

[0033] Figure 3A Representative images of training samples including ICH-stained triple-negative bone marrow aspirate clot samples obtained from AML patients, showing the expression of IRAK4 in the samples. Blasts: 47%.

[0034] Figure 3B Representative images of training samples including ICH-stained triple-negative bone marrow aspirate clot samples obtained from AML patients, showing the expression of NF-κΒ p-p65 S536 in the samples. Blasts: 47%.

[0035] Figure 3C Representative images of training samples including ICH-stained triple-negative bone marrow aspirate clot samples obtained from AML patients, showing the expression of NF-κΒ p-p50 S337 in the samples. Blasts: 47%.

[0036] Figure 4A Representative images of training samples including ICH-stained benign bone marrow aspirate clot samples showing the expression of IRAK4 in the samples. Blasts: 3%.

[0037] Figure 4B Representative images of training samples including ICH-stained benign bone marrow aspirate clot samples showing the expression of NF-κΒ p-p65 S536 in the samples. Blasts: 3%.

[0038] Figure 4C Representative images of training samples including ICH-stained benign bone marrow aspirate clot samples showing the expression of NF-κΒ p-p50 S337 in the samples. Blasts: 3%.

[0039] Figure 5A Representative images of AML training sample ABS04b showing nuclear expression of p-IRAK1 T387.

[0040] Figure 5B Representative images of AML training sample ABS2 showing negative staining for p-IRAK1 T387 expression.

[0041] Figures 6A-6D Representative IHC images of serial sections of FFPE bone marrow aspirate clots obtained from a secondary AML case (training sample 01b), showing quadruple positive nuclear staining for IRAK4 (A), p-IRAK1 T387 (B), NF-κΒ p-p50 S337 (C), and NF-κΒ p-p65 S536 (D).

[0042] Figures 7A-7C Representative IHC images of serial sections of FFPE bone marrow aspirate clots obtained from a progressive AML case showing triple negative staining assays for IRAK4 (A), NF-κΒ p-p50 S337 (B), and NF-κΒ p-p65 S536 (C).

[0043] Figures 8A-8CRepresentative IHC images of serial sections of FFPE bone marrow aspirate clots obtained from an AML case, showing triple positive staining test for IRAK4 (A), NF-κΒ p-p50 S337 (B), and NF-κΒ p-p65 S536 (C).

[0044] Fig. 9 Representative images of triple-positive, IRAK4-cytoplasmic stained tissue sections before and after hematoxylin counterstaining are shown. Sections of AML case samples, FFPE bone marrow aspirate clots.

[0045] Figures 10A-10C Representative images showing cytoplasmic staining of IRAK4 (A), NF-κB p-p50 S337 (B), and NF-κB p-p65 S536 (C) in a triple positive cytoplasmic staining test.

[0046] Fig.11A and 11B Representative images of triple positive (A) or negative (B) nuclear staining tests for MYD88 expression.

[0047] Figures 12A-12D It is a representative image showing the results of staining experiments on OCI-LY3 xenograft tumor (MYD88-mut) tissue sections. The images show the expression of IRAK4 in the nucleus of cells in each sample (A), the expression of MYD-88 in the cytoplasm (B), the expression of NF-κΒ p-p65 in the nucleus (C), and the expression of NF-κΒ p-p50 in the nucleus (D).

[0048] Fig.13A and 13B Representative images of IHC staining showing IRAK4 expression in tissue sections collected from lymphoma xenograft tumors. Nuclear expression (A) and cytoplasmic expression (B) are shown.

[0049] Fig.14A and 14B Representative images of IRAK4 IHC staining in lymphoma training samples, showing samples positive nuclear expression of IRAK4 (A) and negative expression of IRAK4 (B).

[0050] Fig.15A and 15B Representative images of IHC staining in lymphoma tissue sections showing nuclear expression of IRAK4 (A) and NF-κΒ p-p50 (B).

[0051] Fig.16are images showing the results of western blot tests of IRAK4 and MYD88 expression in nuclear extracts of leukemia and lymphoma cell lines. The results of the western blot tests support the observations in the IHC staining experiments.

[0052] Fig.17 Representative images of FLT3 staining in AML bone marrow training samples (blasts: 84%).

[0053] Fig.18 Dose-dependent objective responses are shown for human patients receiving specific doses of Compound 1.

[0054] Fig.19 Schematic representation of the IRAK1 / 4 complex with the adaptor protein MYD88. Upon substrate binding to IL-R1 or TLR, MYD88 activates the recruitment of the IRAK4 / 1 complex, allowing phosphorylation of IRAK-1. Phosphorylated IRAK-1 then binds to TRAF-6, activating NK-kB signaling, leading to inflammation and tumor promotion. The MYD88-L265P mutation results in a sustained upregulation of this pathway. Compound 1 inhibits IRAK4.

[0055] Fig. 20 Dose-dependent objective responses are shown for human patients receiving specific doses of Compound 1.

[0056] Fig.21 The responses of certain human patients receiving Compound 1 are shown.

[0057] Fig.22A -C shows the efficacy of Compound 1 on certain in vivo models of non-Hodgkin's lymphoma. In each case, administration of Compound 1 reduced tumor growth.

[0058] Fig.23 The efficacy of Compound 1 in combination with ibrutinib is shown. The combination of Compound 1 and ibrutinib demonstrated a synergistic reduction in tumor growth compared to Compound 1 or ibrutinib alone.

[0059] Fig.24 The oral pharmacology profile of Compound 1 in humans at an exemplary dose is shown. After oral administration, Compound 1 is rapidly absorbed, with maximum plasma concentrations observed at 0.5 hours to 8 hours after administration. Compound 1 exhibits a dose proportional increase in exposure and has a half-life of approximately 6 hours. After multiple single doses per day, minimal to no accumulation is observed. After multiple twice-daily doses, moderate accumulation is observed at steady state. In summary, the oral pharmacokinetics of Compound 1 are desirable.

[0060] Fig.25The percentage reduction in tumor burden is shown for subjects receiving 300 mg twice daily. Compound 1 had an acceptable safety and tolerability profile at RP2D, including 3 patients who have been on study for 1-2 years.

[0061] Fig.26A Shown are the effects of exemplary concentrations of Compound 1 on erythroid differentiation of primary MDS / AML hematopoietic stem and progenitor cells (HSCP).

[0062] Fig.26B Shown are the effects of exemplary concentrations of Compound 1 on neutrophil differentiation from primary MDS / AML hematopoietic stem and progenitor cells (HSCP).

[0063] Fig.27A Shown is the effect of Compound 1 on spleen weight in leukemic xerographic copies after 6 weeks of treatment at 12.5 mg / kg.

[0064] Fig.27B Shown is the effect of Compound 1 on liver weight in leukemic xerographic copies after 6 weeks of treatment at 12.5 mg / kg.

[0065] Fig.27C The effect of Compound 1 on the % of leukemic cells in the bone marrow in leukemic xerographic copies is shown after 6 weeks of treatment at 12.5 mg / kg. Compound 1 reduces disease burden in THP-1 xerographic copies.

[0066] Fig.28 The design of the study described in Example 4 is described.

[0067] Fig.29A The effect of compound 1 on NF-κB phosphorylated-p50 expression is shown. NF-κB phosphorylated p50 protein expression in pre-treatment biopsies correlates with lymphoma shrinkage or stable disease. SD refers to stable disease, and PD refers to progressive disease.

[0068] Fig.29B The effect of compound 1 on NF-κB phosphorylation-p50 expression is shown. Inhibition of NF-κB phosphorylation-p50 expression indicates that compound 1 is inhibiting IRAK4 and downregulating NF-κB. During treatment with compound 1, NF-κB phosphorylation-p50 expression was inhibited (positive to negative).

[0069] Fig.29C The expression of NF-κB phosphorylation-p50 in human tonsil cells and lymphoma cells is shown. The expression of NF-κB phosphorylation-p50 is increased in lymphoma cells.

[0070] Fig. 30Ais a Western blot of OCL-LY10 cells treated with DMSO or compound 1. Treatment with compound 1 downregulated the expression of NF-κB phospho-p50.

[0071] Fig. 30B is a Western blot of AML cells treated with DMSO or compound 1. Treatment with compound 1 downregulated the expression of NF-κB phosphorylated-p50.

[0072] Fig.31A and 31B Representative images of FFPE BM samples obtained from AML patients showing the expression of NF-κBp-p50 in the samples. DETAILED DESCRIPTION

[0073] Acute myeloid leukemia (AML) is the second most common form of leukemia in adults and remains a highly lethal disease. Interleukin-1 receptor-associated kinase 4 (IRAK4) has been shown to be a potential therapeutic target for human AML. IRAK4-mediated activation of the NF-κB signaling pathway may play a key role in NF-κB-regulated cancer cell survival and chemoresistance. Results from an ongoing Phase 1 study demonstrate that the IRAK4 inhibitor emavusertib (compound 1) has clinical activity in patients with relapsed / refractory AML and high-risk MDS. To support the development of a companion diagnostic for emavusertib, we developed an immunohistochemistry (IHC) assay and explored the expression of potential biomarkers in bone marrow (BM) samples obtained from AML patients. The present disclosure relates to methods for analyzing the expression of IRAK4, NF-κB p-p50, and NF-κB p-p65 in human AML (including IHC staining), and further evaluating these molecules as potential biomarkers for emavusertib therapy in AML patients.

[0074] In some embodiments of the present disclosure, we use IHC staining and immunoblotting to determine the expression of IRAK4, NF-κB p-p50 S337, and NF-κB p-p65 S536 proteins in human leukemia cell lines and clinical AML samples. Exploratory biomarker evaluation was performed using serial sections of formalin-fixed paraffin-embedded BM clot samples obtained from 19 AML patients. Eight BM AML samples were purchased from Analytical Biological Services, and 11 BM samples were obtained from evaluable relapsed / refractory AML patients during screening. Using IHC staining, we found that 9 / 19 AML cases had IRAK4 nuclear expression in the mother cells. To our knowledge, this is the first report showing nuclear accumulation of IRAK4 in cancer cells. To support our findings in clinical AML samples, we detected IRAK4 protein expression in nuclear lysates prepared from leukemia cell lines THP-1, HL-60, and K562. Using AML BM samples, we found that IRAK4 nuclear expression in blasts was significantly associated with activation of NF-κB, as determined by nuclear accumulation of NF-κB p-p50 and p-p65 in 9 / 19 cases. Cytoplasmic expression of IRAK4 was detected in 2 / 19 AMLs, whereas IRAK4, NF-κB p-p50, and p-p65 expression were not detected in 8 / 19 AMLs. Clinical response data will be presented in the context of these novel findings.

[0075] Our findings reveal a previously unknown nuclear expression of IRAK4 in leukemic cells. Our results demonstrate that nuclear IRAK4, NF-κB p-p50, and p-p65 are co-expressed in blasts, suggesting a possible novel mode of interaction between IRAK4 and NF-κB in human AML. Although the role of nuclear IRAK4 in leukemic cells remains to be investigated, our preliminary findings reveal a new perspective for imatinib therapy stratification and demonstrate the potential for novel biomarker discovery through IHC analysis of clinical samples.

[0076] Activation of IRAK4 leads to activation of the NF-κB signaling pathway, including phosphorylation of NF-κB p50, which is required for DNA binding and transcriptional activity of NF-κB (Hou S et al. Phosphorylation of serine 337 of NF-κB p50 is critical for DNA binding. J Biol Chem. 2003). Phosphorylation of S536 in the NF-κB p65 transcriptional activation domain (TAD) leads to enhanced transcriptional activation by increasing CBP / p300 binding and acetylation at K310 of p65.

[0077] In certain aspects, the present disclosure provides a method of treating a disease or disorder in a subject, the method comprising:

[0078] obtaining one or more biological samples from a subject;

[0079] analyzing the biological sample for the presence of a first biomarker in nuclei of cells of the biological sample; and

[0080] administering to the subject an IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader when the presence of the first biomarker is detected in the nucleus of the cell of the biological sample;

[0081] The first biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

[0082] In certain embodiments, the methods of the present disclosure further comprise:

[0083] analyzing the biological sample for the presence of a second biomarker in nuclei of cells of the biological sample;

[0084] administering an IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader to the subject only when the presence of the first biomarker and the second biomarker is detected in the nucleus of the cell of the biological sample; and

[0085] Wherein the second biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

[0086] In some embodiments, the methods of the present disclosure further include:

[0087] analyzing the biological sample for the presence of a third biomarker in nuclei of cells of the biological sample; and

[0088] administering to the subject an IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader only when the presence of the first biomarker, the second biomarker, and the third biomarker is detected in the nucleus of the cell of the biological sample;

[0089] wherein the third biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

[0090] In certain embodiments, the methods of the present disclosure further comprise:

[0091] analyzing the biological sample for the presence of a fourth biomarker in nuclei of cells of the biological sample; and

[0092] administering an IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader to the subject only when the presence of the first biomarker, the second biomarker, the third biomarker, and the fourth biomarker is detected in the nucleus of the cell of the biological sample;

[0093] wherein the fourth biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

[0094] In some aspects, the present disclosure provides a method of treating a disease or disorder in a subject, the method comprising:

[0095] obtaining one or more biological samples from the subject;

[0096] measuring a sample expression level of a first biomarker in the biological sample;

[0097] comparing the sample expression level of the first biomarker to a reference expression level of the first biomarker;

[0098] administering to the subject an IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader when the sample expression level of the first biomarker in the sample is elevated compared to the reference expression level of the first biomarker; and

[0099] The first biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

[0100] In certain embodiments, the methods of the present disclosure further comprise:

[0101] measuring the sample expression level of a second biomarker in the biological sample;

[0102] comparing the sample expression level of the second biomarker to a reference expression level of the second biomarker; and

[0103] administering to said subject an IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader only if said sample expression levels of said first and second biomarkers in said sample are both elevated as compared to said reference expression levels of said first and second biomarkers;

[0104] The second biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

[0105] In some embodiments, the methods of the present disclosure further include:

[0106] measuring the sample expression level of a third biomarker in the biological sample;

[0107] comparing the sample expression level of the third biomarker to a reference expression level of the third biomarker; and

[0108] administering an IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader to the subject only if the sample expression levels of the first biomarker, the second biomarker, and the third biomarker in the sample are each elevated compared to the reference expression levels of the first biomarker, the second biomarker, and the third biomarker;

[0109] wherein the third biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

[0110] In certain embodiments, the methods of the present disclosure further comprise:

[0111] measuring the sample expression level of a fourth biomarker in the biological sample;

[0112] comparing the sample expression level of the fourth biomarker to a reference expression level of the fourth biomarker; and

[0113] administering an IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader to the subject only if the sample expression levels of the first biomarker, the second biomarker, the third biomarker, and the fourth biomarker in the sample are each elevated compared to the reference expression levels of the first biomarker, the second biomarker, the third biomarker, and the fourth biomarker;

[0114] wherein the fourth biomarker is selected from IRAK4, NF-κBp-p65, NF-κBp-p50, p-IRAK1, FLT3 and MYD88.

[0115] In other aspects, the present disclosure provides methods of treating a disease or condition in a subject, the method comprising:

[0116] obtaining one or more biological samples from the subject;

[0117] measuring a sample expression level of a first biomarker in the biological sample;

[0118] comparing the sample expression level of the first biomarker to a reference expression level of the first biomarker;

[0119] analyzing the biological sample for the presence of the first biomarker in nuclei of cells of the biological sample; and

[0120] An IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader is administered to the subject only if:

[0121] detecting the presence of the first biomarker in the nucleus of the cell of the biological sample; and

[0122] the sample expression level of the first biomarker in the sample is increased compared to the reference expression level of the first biomarker;

[0123] The first biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

[0124] In certain embodiments, the methods of the present disclosure further comprise:

[0125] measuring the sample expression level of a second biomarker in the biological sample;

[0126] comparing the sample expression level of the second biomarker to a reference expression level of the second biomarker;

[0127] analyzing the biological sample for the presence of the second biomarker in nuclei of cells of the biological sample; and

[0128] An IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader is administered to the subject only if:

[0129] detecting the presence of both the first biomarker and the second biomarker in the nucleus of the cell of the biological sample; and

[0130] the sample expression levels of the first biomarker and the second biomarker in the sample are both increased compared to the reference expression levels of the first biomarker and the second biomarker;

[0131] Wherein the second biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

[0132] In some embodiments, the methods of the present disclosure further include:

[0133] measuring the sample expression level of a third biomarker in the biological sample;

[0134] comparing the sample expression level of the third biomarker to a reference expression level of the third biomarker;

[0135] analyzing the biological sample for the presence of the third biomarker in nuclei of cells of the biological sample; and

[0136] An IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader is administered to the subject only if:

[0137] detecting the presence of each of the first biomarker, the second biomarker, and the third biomarker in the nucleus of the cell of the biological sample; and

[0138] the sample expression level of each of the first biomarker, the second biomarker, and the third biomarker in the sample is elevated compared to the reference expression level of each of the first biomarker, the second biomarker, and the third biomarker;

[0139] wherein the third biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

[0140] In certain embodiments, the methods of the present disclosure further comprise:

[0141] measuring the sample expression level of a fourth biomarker in the biological sample;

[0142] comparing the sample expression level of the fourth biomarker to a reference expression level of the fourth biomarker;

[0143] analyzing the biological sample for the presence of the fourth biomarker in nuclei of cells of the biological sample; and

[0144] An IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader is administered to the subject only if:

[0145] detecting the presence of each of the first biomarker, the second biomarker, the third biomarker, and the fourth biomarker in the nucleus of the cell of the biological sample; and

[0146] the sample expression level of each of the first biomarker, the second biomarker, the third biomarker, and the fourth biomarker in the sample is elevated compared to the reference expression level of each of the first biomarker, the second biomarker, the third biomarker, and the fourth biomarker;

[0147] wherein the fourth biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

[0148] In certain embodiments of the present disclosure, the first biomarker is IRAK4. In other embodiments, the first biomarker is NF-κB p-p65. In other embodiments, the first biomarker is NF-κB p-p50. In certain embodiments, the first biomarker is p-IRAK1. In other embodiments, the first biomarker is FLT3. In other embodiments, the first biomarker is MYD88.

[0149] In certain embodiments, the second biomarker is IRAK4. In other embodiments, the second biomarker is NF-κB p-p65. In other embodiments, the second biomarker is NF-κB p-p50. In certain embodiments, the second biomarker is p-IRAK1. In other embodiments, the second biomarker is FLT3. In other embodiments, the second biomarker is MYD88.

[0150] In certain embodiments, the third biomarker is present in the sample. In certain such embodiments, when present, the third biomarker is IRAK4. In other embodiments, when present, the third biomarker is NF-κB p-p65. In other embodiments, when present, the third biomarker is NF-κB p-p50. In certain embodiments, the third biomarker is p-IRAK1. In other embodiments, when present, the third biomarker is FLT3. In other embodiments, when present, the third biomarker is MYD88.

[0151] In certain embodiments, the fourth biomarker is present in the sample. In certain such embodiments, when present, the fourth biomarker is IRAK4. In other embodiments, when present, the fourth biomarker is NF-κB p-p65. In other embodiments, when present, the fourth biomarker is NF-κB p-p50. In certain embodiments, the fourth biomarker is p-IRAK1. In other embodiments, when present, the fourth biomarker is FLT3. In other embodiments, when present, the fourth biomarker is MYD88.

[0152] In some embodiments, the sample expression level of the first biomarker indicates the nuclear expression amount of the first biomarker, and the corresponding reference expression level of the first biomarker represents the nuclear expression amount of the first biomarker. In other embodiments, the sample expression level of the first biomarker indicates the cytoplasmic expression amount of the first biomarker, and the corresponding reference expression level of the first biomarker represents the cytoplasmic expression amount of the first biomarker. In some embodiments, the sample expression level of the first biomarker indicates the membrane expression amount of the first biomarker, and the corresponding reference expression level of the first biomarker represents the membrane expression amount of the first biomarker.

[0153] In some embodiments, the sample expression level of the second biomarker represents the nuclear expression amount of the second biomarker, and the reference expression level of the second biomarker indicates the nuclear expression amount of the second biomarker. In some embodiments, the sample expression level of the second biomarker represents the cytoplasmic expression amount of the second biomarker, and the reference expression level of the second biomarker indicates the cytoplasmic expression amount of the second biomarker. In some embodiments, the sample expression level of the second biomarker represents the membrane expression amount of the second biomarker, and the reference expression level of the second biomarker indicates the membrane expression amount of the second biomarker.

[0154] In some embodiments, when present, the sample expression level of the third biomarker represents the nuclear expression amount of the third biomarker, and when present, the reference expression level of the third biomarker indicates the nuclear expression amount of the third biomarker. In other embodiments, when present, the sample expression level of the third biomarker represents the cytoplasmic expression amount of the third biomarker, and when present, the reference expression level of the third biomarker indicates the cytoplasmic expression amount of the third biomarker. In other embodiments, when present, the sample expression level of the third biomarker represents the membrane expression amount of the third biomarker, and when present, the reference expression level of the third biomarker indicates the membrane expression amount of the third biomarker.

[0155] In some embodiments, when present, the sample expression level of the fourth biomarker indicates the nuclear expression of the fourth biomarker, and when present, the reference expression level of the fourth biomarker indicates the nuclear expression of the fourth biomarker. In other embodiments, when present, the sample expression level of the fourth biomarker indicates the cytoplasmic expression of the fourth biomarker, and when present, the reference expression level of the fourth biomarker indicates the cytoplasmic expression of the fourth biomarker. In other embodiments, when present, the sample expression level of the fourth biomarker indicates the membrane expression of the fourth biomarker, and when present, the reference expression level of the fourth biomarker indicates the membrane expression of the fourth biomarker.

[0156] In some embodiments, when present, the sample expression level of IRAK4 indicates the nuclear expression amount of IRAK4, and when present, the reference expression level of IRAK4 indicates the nuclear expression amount of IRAK4. In other embodiments, when present, the sample expression level of IRAK4 indicates the cytoplasmic expression amount of IRAK4, and when present, the reference expression level of IRAK4 indicates the cytoplasmic expression amount of IRAK4.

[0157] In some embodiments, when present, the sample expression level of NF-κB p-p65 indicates the nuclear expression amount of NF-κB p-p65. In certain embodiments, when present, the reference expression level of NF-κB p-p65 indicates the nuclear expression amount of NF-κB p-p65. In some embodiments, when present, the sample expression level of NF-κB p-p65 indicates the cytoplasmic expression amount of NF-κB p-p65. In certain embodiments, when present, the reference expression level of NF-κB p-p65 indicates the cytoplasmic expression amount of NF-κB p-p65.

[0158] In some embodiments, when present, the sample expression level of NF-κB p-p50 indicates the nuclear expression amount of NF-κB p-p50. In certain embodiments, when present, the reference expression level of NF-κB p-p50 indicates the nuclear expression amount of NF-κB p-p50. In some embodiments, when present, the sample expression level of NF-κB p-p50 indicates the cytoplasmic expression amount of NF-κB p-p50. In certain embodiments, when present, the reference expression level of NF-κB p-p50 indicates the cytoplasmic expression amount of NF-κB p-p50.

[0159] In some embodiments, when present, the sample expression level of p-IRAK1 indicates the nuclear expression amount of p-IRAK1, and when present, the reference expression level of p-IRAK1 indicates the nuclear expression amount of p-IRAK1. In other embodiments, when present, the sample expression level of p-IRAK1 indicates the cytoplasmic expression amount of p-IRAK1, and when present, the reference expression level of p-IRAK1 indicates the cytoplasmic expression amount of p-IRAK1.

[0160] In some embodiments, when present, the sample expression level of FLT3 indicates the cytoplasmic expression of FLT3. In certain embodiments, when present, the reference expression level of FLT3 indicates the cytoplasmic expression of FLT3. In some embodiments, when present, the sample expression level of FLT3 indicates the membrane expression of FLT3. In certain embodiments, when present, the reference expression level of FLT3 indicates the membrane expression of FLT3.

[0161] In some embodiments, when present, the sample expression level of MYD88 indicates the nuclear expression amount of MYD88. In certain embodiments, when present, the reference expression level of MYD88 indicates the nuclear expression amount of MYD88. In some embodiments, when present, the sample expression level of MYD88 indicates the cytoplasmic expression amount of MYD88. In certain embodiments, when present, the reference expression level of MYD88 indicates the cytoplasmic expression amount of MYD88.

[0162] In some embodiments, the reference expression level is a value representing or obtained from a subject or subjects not suffering from a disease or condition. In further embodiments, the value of the reference expression level is obtained from tissue or blood. In further embodiments, the value is obtained from blood.

[0163] In certain embodiments, the IRAK4 modifying compound is an IRAK4 inhibitor. In other embodiments, the IRAK4 modifying compound is an IRAK4 degrader.

[0164] Methods and compounds related to the contents of the present disclosure can be found, for example, in U.S. Patent Nos. 10,160,753, 9,732,095, 10,758,518, published U.S. patent application No.: 20210290628, U.S. application No.: 17 / 680,995, and PCT application Nos. PCT / WO2022 / 031330 and PCT / US2021 / 59668; the contents of which are incorporated herein by reference in their entirety, particularly with respect to the IRAK4 inhibitors, treatment regimens, and disease indications disclosed therein.

[0165] IRAK4 inhibitors

[0166] In a broad sense, the methods disclosed herein can be performed with any IRAK4 inhibitor. For example, the methods can be performed with IRAK4 inhibitors disclosed in PCT / IB2015 / 050119, PCT / IB2015 / 050217, PCT / IB2015 / 0054620, PCT / IB2016 / 054203 and / or PCT / IB2016 / 054229. The contents of each of the above international applications are incorporated herein by reference in their entirety, particularly the IRAK4 inhibitors disclosed therein.

[0167] In certain embodiments, the IRAK4 inhibitor is of Formula I:

[0168]

[0169] or a pharmaceutically acceptable salt thereof;

[0170] in

[0171] X1 and X3 are independently CH or N; X2 is CR2 or N; wherein one and not more than one of X1, X2 or X3 is N;

[0172] A is O or S;

[0173] Y is -CH2- or O;

[0174] Z is an aryl group or a heterocyclic group;

[0175] R1 is independently at each occurrence halo or optionally substituted heterocyclyl; wherein the substituent is alkyl, alkoxy, aminoalkyl, halo, hydroxy, hydroxyalkyl or -NR a R b ;

[0176] R2 is hydrogen, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl or -NR a R b ; wherein the substituent is an alkyl, amino, halide or hydroxyl group;

[0177] R3, at each occurrence, is alkyl or hydroxy;

[0178] R a and R b are independently hydrogen, alkyl, acyl or heterocyclyl;

[0179] 'm' and 'n' are independently 0, 1 or 2;

[0180] 'p' is either 0 or 1.

[0181] In certain embodiments, A is O or S; Y is -CH2- or O; Z is aryl or heterocyclyl; R1 at each occurrence is independently halo or optionally substituted heterocyclyl, wherein the substituent is alkyl, aminoalkyl, halo or -NR a R b ; where R a and R b are independently hydrogen, alkyl or heterocyclyl; R2 is hydrogen, cycloalkyl, heterocyclyl or -NR a R b ; 'm' is 0; and 'n' is 1.

[0182] In other embodiments, A is O or S; Y is -CH2- or O; Z is aryl or heterocyclyl; R1 at each occurrence is independently halo or optionally substituted heterocyclyl; wherein the substituent is alkyl, alkoxy, aminoalkyl, halo, hydroxy or -NR a R b ; where R a and R b R2 is hydrogen, cycloalkyl, optionally substituted heterocyclyl or -NR a R b , wherein the substituents are selected from amino, halo or hydroxy; 'm' and 'n' are independently 0, 1 or 2; and 'p' is 0 or 1.

[0183] In certain embodiments, yes

[0184]

[0185] In certain embodiments, Z is aryl or a 5- or 6-membered heterocyclyl. In certain embodiments, Z is an optionally substituted heterocyclyl selected from phenyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, 1H-tetrazolyl, oxadiazolyl, triazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, azetidinyl, oxetanyl, imidazolidinyl, pyrrolidinyl, oxazolidinyl, thiazolidinyl, pyrazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, 1,4-dioxanyl, dioxythiomorpholinyl, oxapiperazinyl, oxapiperidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl, dihydropyranyl, and azabicyclo[3.2.1]octyl; each of which is optionally substituted with alkyl, alkoxy, halo, hydroxy, hydroxyalkyl, or -NR a R b Replace; and R a and R b are independently hydrogen, alkyl or acyl.

[0186] In certain embodiments, the IRAK4 inhibitor is of formula (IA):

[0187]

[0188]

[0189] or a pharmaceutically acceptable salt thereof. In certain embodiments, A is O or S; Y is -CH2- or O; R1 is independently halo or optionally substituted heterocyclyl at each occurrence, wherein the substituent is alkyl, aminoalkyl, halo or -NR a R b ; where R a and R b are independently hydrogen, alkyl or heterocyclyl; R2 is hydrogen, cycloalkyl, heterocyclyl or -NR a R b ; 'm' is 0; and 'n' is 1. In other embodiments, A is O or S; Y is -CH2- or O; R1 is independently at each occurrence halo or optionally substituted heterocyclyl; wherein the substituent is alkyl, alkoxy, aminoalkyl, halo, hydroxy or -NR a R b ; where R a and R b R2 is hydrogen, cycloalkyl, optionally substituted heterocyclyl or -NR a R b , wherein the substituents are selected from amino, halo or hydroxy; and 'm' and 'n' are independently 0, 1 or 2.

[0190] In certain embodiments, the IRAK4 inhibitor is of formula (IB):

[0191]

[0192] or a pharmaceutically acceptable salt thereof. In certain embodiments, A is O or S; Y is -CH2- or O; R1 is independently halo or optionally substituted heterocyclyl at each occurrence, wherein the substituent is alkyl, aminoalkyl, halo or -NR a R b ; where R a and R b are independently hydrogen, alkyl or heterocyclyl; R2 is hydrogen, cycloalkyl, heterocyclyl or -NR a R b ; and 'n' is 1. In other embodiments, A is O or S; Y is -CH2- or O; R1 is independently at each occurrence halo or optionally substituted heterocyclyl; wherein the substituent is alkyl, alkoxy, aminoalkyl, halo, hydroxy or -NR a R b; where R a and R b R2 is hydrogen, cycloalkyl, optionally substituted heterocyclyl or -NR a R b , wherein the substituents are selected from amino, halo or hydroxy; and 'm' and 'n' are independently 0, 1 or 2.

[0193] In certain embodiments, the IRAK4 inhibitor is of formula (IC):

[0194]

[0195] or a pharmaceutically acceptable salt thereof.

[0196] In certain embodiments, R1 is optionally substituted heterocyclyl; wherein the substituent is alkyl, alkoxy, aminoalkyl, halo, hydroxy, hydroxyalkyl, or -NR a R b ; and R a and R b is independently hydrogen or acyl. In other embodiments, R1 is an optionally substituted heterocyclyl; wherein the substituent is alkyl, aminoalkyl, halo or -NR a R b ; and R a and R b is independently hydrogen or acyl. In other embodiments, R1 is an optionally substituted heterocyclyl; the substituent is alkyl, alkoxy, aminoalkyl, halo, hydroxyl or -NR a R b ; where R a and R b R1 is independently hydrogen, alkyl or heterocyclyl. In certain embodiments, R1 is pyridyl, pyrazolyl, pyrrolidinyl or piperidinyl. In certain embodiments, R1 is optionally substituted pyrazolyl, wherein the substituent is alkyl, hydroxyl or -NR a R b In other embodiments, R1 is halo.

[0197] In certain embodiments, R2 is hydrogen, cycloalkyl, optionally substituted heterocyclyl, or -NR a R b , wherein the substituent is selected from amino, halo or hydroxy. In certain embodiments, R2 is hydrogen, cycloalkyl, optionally substituted heterocyclyl or -NR a R b, wherein the substituent is selected from amino, halo or hydroxy. In certain embodiments, R2 is an optionally substituted heterocyclic group selected from the following: piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, azetidinyl, pyrazolyl, furanyl or azabicyclo[3.2.1]octyl; wherein the substituent is hydroxy, halo, alkyl or amino. In certain embodiments, R2 is piperidinyl, pyrrolidinyl, morpholinyl or piperazinyl. In other embodiments, R2 is hydrogen. In other embodiments, is cycloalkyl. In certain embodiments, R2 is cyclopropyl.

[0198] In certain embodiments, R3 is alkyl.

[0199] In certain embodiments, m is 0 and p is 1. In other embodiments, m is 0 or 2 and p is 0 or 1.

[0200] In certain embodiments, the IRAK4 inhibitor is selected from:

[0201] 6'-amino-N-(2-morpholino-oxazolo[4,5-b]pyridin-6-yl)-[2,3'-bipyridine]-6-carboxamide;

[0202] 6'-amino-N-(5-cyclopropyl-2-morpholinyloxazolo[4,5-b]pyridin-6-yl)-[2,3'-bipyridine]-6-carboxamide hydrochloride;

[0203] N-(5-cyclopropyl-2-morpholinyloxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide hydrochloride;

[0204] N-(2,5-di(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)-6-(1H-pyrazol-4-yl)picolinamide hydrochloride;

[0205] N-(2,5-di(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0206] N-(2-morpholinyl-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)-6-(1H-pyrazol-4-yl)picolinamide;

[0207] 2-(2-methylpyridin-4-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0208] 6-Chloro-N-(2-morpholinyl-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)picolinamide;

[0209] N-(2,5-di(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)-6-(1-methyl-1H-pyrazol-4-yl)picolinamide;

[0210] 2-(2-chloropyridin-4-yl)-N-(2,5-di(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0211] (S)-2-(2-methylpyridin-4-yl)-N-(2-morpholinyl-5-(pyrrolidin-3-ylamino)oxazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0212] 6'-amino-N-(2-morpholino-oxazolo[5,4-b]pyridin-5-yl)-[2,3'-bipyridine]-6-carboxamide;

[0213] 6'-amino-N-(2-morpholinothiazolo[4,5-c]pyridin-6-yl)-[2,3'-bipyridine]-6-carboxamide;

[0214] 6'-amino-N-(2-morpholinothiazolo[5,4-b]pyridin-5-yl)-[2,3'-bipyridine]-6-carboxamide;

[0215] 2-(2-methylpyridin-4-yl)-N-(2-morpholinothiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0216] 6'-amino-N-(2-morpholinothiazolo[4,5-b]pyridin-6-yl)-[2,3'-bipyridine]-6-carboxamide;

[0217] N-(2-morpholinothiazolo[4,5-b]pyridin-6-yl)-6-(1H-pyrazol-4-yl)picolinamide;

[0218] 3-(4-(Aminomethyl)piperidin-1-yl)-5-fluoro-N-(2-morpholinothiazolo[4,5-b]pyridin-6-yl)benzamide;

[0219] 2-(4-(Aminomethyl)piperidin-1-yl)-5-fluoro-N-(2-morpholinothiazolo[4,5-b]pyridin-6-yl)benzamide;

[0220] 2-(2-methylpyridin-4-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0221] N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)-6-(1H-pyrazol-4-yl)picolinamide;

[0222] N-(2,5-di(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)-6-(1H-pyrazol-4-yl)picolinamide;

[0223] N-(2,5-di(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0224] N-(2,5-dimorpholinyloxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0225] N-(5-(4-methylpiperazin-1-yl)-2-morpholinooxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0226] N-(2,5-di(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)-2-(6-methoxypyridin-3-yl)oxazole-4-carboxamide;

[0227] N-(2,5-di(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-3-yl)oxazole-4-carboxamide;

[0228] N-(2,5-di(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)-2-(2-hydroxypyridin-3-yl)oxazole-4-carboxamide;

[0229] 2-(2-Hydroxypyridin-3-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0230] N-(2,5-di(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)-2-(6-hydroxypyridin-3-yl)oxazole-4-carboxamide;

[0231] 2-(2-methoxypyridin-4-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0232] 2-(2-methylpyridin-3-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0233] 2-(3-methylpyridin-4-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0234] N-(2,5-di(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)-2-(3-methylpyridin-4-yl)oxazole-4-carboxamide;

[0235] 2-(6-methylpyridin-3-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0236] 6-(1-methyl-1H-pyrazol-4-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)picolinamide;

[0237] N-(2,5-di(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)-2-(6-methylpyridin-3-yl)oxazole-4-carboxamide;

[0238] (S)-N-(5-(3-aminopyrrolidin-1-yl)-2-morpholinyloxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0239] (S)-N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinyloxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0240] (R)-N-(5-(3-aminopyrrolidin-1-yl)-2-morpholinooxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0241] (R)-N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinyloxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0242] (S)-2-(3-aminopyrrolidin-1-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0243] (S)-6-(3-hydroxypyrrolidin-1-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)picolinamide;

[0244] (S)-6-(3-aminopyrrolidin-1-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)picolinamide;

[0245] (S)-2-(3-hydroxypyrrolidin-1-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0246] (S)-N-(5-cyclopropyl-2-morpholinyloxazolo[4,5-b]pyridin-6-yl)-2-(3-hydroxypyrrolidin-1-yl)oxazole-4-carboxamide;

[0247] (S)-2-(3-aminopyrrolidin-1-yl)-N-(5-cyclopropyl-2-morpholinyloxazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0248] 2-(2-methylpyridin-4-yl)-N-(5-(piperidin-1-yl)-2-(pyrrolidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide hydrochloride;

[0249] N-(2-(2,6-dimethylmorpholinyl)-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide hydrochloride;

[0250] N-(2,5-di(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)-6-(1-methyl-1H-pyrazol-4-yl)picolinamide hydrochloride;

[0251] 6-(1-methyl-1H-pyrazol-4-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)picolinamide;

[0252] N-(2,5-di(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-3-yl)oxazole-4-carboxamide hydrochloride;

[0253] N-(2-((2S,6R)-2,6-dimethylmorpholinyl)-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0254] 2-(2-methylpyridin-3-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0255] 2-(2-Hydroxypyridin-3-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0256] N-(2,5-di(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)-2-(2-methoxypyridin-4-yl)oxazole-4-carboxamide;

[0257] 2-(6-methoxypyridin-3-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0258] 2-(2-methoxypyridin-4-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0259] (S)-N-(5-(3-fluoropiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0260] 2-(6-methylpyridin-3-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0261] 2-(3-methylpyridin-4-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0262] (S)-6-(3-aminopyrrolidin-1-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)picolinamide;

[0263] (S)-6-(3-hydroxypyrrolidin-1-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)picolinamide;

[0264] (S)-6-(3-aminopyrrolidin-1-yl)-N-(2,5-di(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)picolinamide;

[0265] (S)-N-(2,5-di(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)-6-(3-hydroxypyrrolidin-1-yl)picolinamide;

[0266] (S)-2-(3-aminopyrrolidin-1-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0267] (S)-N-(5-(3-aminopyrrolidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0268] (S)-2-(3-aminopyrrolidin-1-yl)-N-(5-cyclopropyl-2-morpholinothiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0269] N-(5-cyclopropyl-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0270] (S)-2-(3-hydroxypyrrolidin-1-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0271] (S)-N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0272] (S)-N-(5-cyclopropyl-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-6-(3-hydroxypyrrolidin-1-yl)picolinamide;

[0273] (S)-N-(5-cyclopropyl-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(3-hydroxypyrrolidin-1-yl)oxazole-4-carboxamide;

[0274] (S)-N-(5-cyclopropyl-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-6-(1-(2-hydroxypropyl)-1H-pyrazol-4-yl)picolinamide;

[0275] (S)-N-(5-cyclopropyl-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(1-(2-hydroxypropyl)-1H-pyrazol-4-yl)oxazole-4-carboxamide;

[0276] N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(6-methoxypyridin-3-yl)oxazole-4-carboxamide;

[0277] (S)-N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(6-methoxypyridin-3-yl)oxazole-4-carboxamide;

[0278] (R)-N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(6-methoxypyridin-3-yl)oxazole-4-carboxamide;

[0279] (S)-N-(5-(azetidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-6-(3-hydroxypyrrolidin-1-yl)picolinamide;

[0280] N-(5-(3-hydroxyazetidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0281] (S)-N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-5-(2-methylpyridin-4-yl)thiophene-2-carboxamide;

[0282] (S)-N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-5-(2-methylpyridin-4-yl)furan-2-carboxamide;

[0283] (S)-N-(5-(3-hydroxypiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0284] N-(5-(4-Hydroxypiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide

[0285] (R)-N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0286] N-(5-(4-hydroxypiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-5-(2-methylpyridin-4-yl)furan-2-carboxamide;

[0287] N-(5-(azetidin-1-yl)-2-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0288] 2-(2-methylpyridin-4-yl)-N-(2-(piperidin-1-yl)-5-(pyrrolidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0289] 2-(2-methylpyridin-4-yl)-N-(2-morpholinyl-5-(pyrrolidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0290] 5-(2-methylpyridin-4-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)furan-2-carboxamide;

[0291] N-(5-(azepan-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0292] 2-(2-aminopyridin-4-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide hydrochloride;

[0293] N-(5-(azetidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0294] (R)-N-(5-(3-hydroxypiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0295] (R)-N-(5-(3-hydroxypiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-5-(2-methylpyridin-4-yl)furan-2-carboxamide;

[0296] (S)-6-(1-(2-hydroxypropyl)-1H-pyrazol-4-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)picolinamide

[0297] N-(5-(4-fluoropiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-5-(2-methylpyridin-4-yl)furan-2-carboxamide

[0298] N-(5-(4-Fluoropiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide hydrochloride

[0299] N-(5-(1-methyl-1H-pyrazol-4-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0300] N-(5-(3-fluorophenyl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0301] N-(5-(4-hydroxypiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-5-(2-methylpyridin-4-yl)furan-2-carboxamide;

[0302] N-(5-(3-fluoropiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-5-(2-methylpyridin-4-yl)furan-2-carboxamide;

[0303] (S)-N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinyloxazolo[4,5-b]pyridin-6-yl)-2-(6-methoxypyridin-3-yl)oxazole-4-carboxamide;

[0304] N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinooxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0305] (R)-N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinyloxazolo[4,5-b]pyridin-6-yl)-2-(6-methoxypyridin-3-yl)oxazole-4-carboxamide;

[0306] N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinyloxazolo[4,5-b]pyridin-6-yl)-2-(6-methoxypyridin-3-yl)oxazole-4-carboxamide;

[0307] (S)-N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinooxazolo[4,5-b]pyridin-6-yl)-5-(2-methylpyridin-4-yl)furan-2-carboxamide;

[0308] (S)-N-(5-(3-hydroxypyrrolidin-1-yl)-2-morpholinooxazolo[4,5-b]pyridin-6-yl)-5-(2-methylpyridin-4-yl)thiophene-2-carboxamide;

[0309] N-(5-(azetidin-1-yl)-2-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0310] 2-(2-methylpyridin-4-yl)-N-(2-(piperidin-1-yl)-5-(pyrrolidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0311] 5-(2-methylpyridin-4-yl)-N-(2-morpholinyl-5-(piperidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)furan-2-carboxamide;

[0312] N-(5-(azetidin-1-yl)-2-morpholinooxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0313] 2-(2-methylpyridin-4-yl)-N-(2-morpholinyl-5-(pyrrolidin-1-yl)oxazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0314] N-(5-(4-hydroxypiperidin-1-yl)-2-morpholinooxazolo[4,5-b]pyridin-6-yl)-5-(2-methylpyridin-4-yl)furan-2-carboxamide;

[0315] (R)-N-(5-(3-hydroxypiperidin-1-yl)-2-morpholinooxazolo[4,5-b]pyridin-6-yl)-5-(2-methylpyridin-4-yl)furan-2-carboxamide;

[0316] N-(5-(Furan-3-yl)-2-morpholinooxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0317] N-(5-(3-fluoropiperidin-1-yl)-2-morpholinooxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0318] N-(5-(4-hydroxypiperidin-1-yl)-2-morpholinooxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0319] N-(5-(4-fluoropiperidin-1-yl)-2-morpholinooxazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0320] (S)-N-(5-(3-aminopiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0321] 2-(2-methylpyridin-4-yl)-N-(2-morpholinyl-5-(1H-pyrazol-4-yl)thiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0322] N-(5-(6-fluoropyridin-3-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0323] N-(5-(3-hydroxy-8-azabicyclo[3.2.1]octan-8-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0324] N-(2-(3-hydroxypiperidin-1-yl)-5-(piperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0325] 2-(2-acetamidopyridin-4-yl)-N-(5-(4-hydroxypiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0326] N-(2-(3-hydroxypiperidin-1-yl)-5-(4-hydroxypiperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0327] 2-(2-acetamidopyridin-4-yl)-N-(5-(3-hydroxypiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide;

[0328] 2-(2-aminopyridin-4-yl)-N-(5-(3-hydroxypiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide hydrochloride;

[0329] 5-(2-aminopyridin-4-yl)-N-(5-(4-hydroxypiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)furan-3-carboxamide hydrochloride;

[0330] 2-(2-aminopyridin-4-yl)-N-(5-(4-hydroxypiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide hydrochloride;

[0331] 2-(2-aminopyridin-4-yl)-N-(5-(4-fluoropiperidin-1-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)oxazole-4-carboxamide hydrochloride;

[0332] N-(5-(2-fluoropyridin-4-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0333] N-(5-(4-fluoropiperidin-1-yl)-2-(3-hydroxypiperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide;

[0334] N-(5-(4-aminopiperidin-1-yl)-2-(3-hydroxypiperidin-1-yl)thiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide hydrochloride; and

[0335] N-(5-(2-hydroxypyridin-4-yl)-2-morpholinothiazolo[4,5-b]pyridin-6-yl)-2-(2-methylpyridin-4-yl)oxazole-4-carboxamide hydrochloride;

[0336] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0337] In certain embodiments, the IRAK4 inhibitor is

[0338] In certain preferred embodiments, the IRAK 4 inhibitor is In other preferred embodiments, the I RAK4 inhibitor is

[0339] Compound 1 can be administered in any amount or manner that causes a desired response in a subject. For example, 100 mg-400 mg of Compound 1 can be administered to a subject twice a day, or 200 mg-1000 mg of Compound 1 can be administered to a subject once a day. In certain embodiments, 100 mg-400 mg of Compound 1 is administered to a subject twice a day. In certain embodiments, 200 mg-400 mg of Compound 1 is administered to a subject twice a day. In certain preferred embodiments, 250 mg-350 mg of Compound 1 is administered to a subject twice a day. In certain embodiments, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, or about 500 mg of Compound 1 is administered to a subject twice a day. In certain embodiments, about 50 mg, about 75 mg, about 100 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, or about 400 mg of Compound 1 is administered to a subject twice a day. In certain embodiments, about 50 mg, about 100 mg, about 200 mg, or about 300 mg of Compound 1 is administered to a subject twice a day. In certain embodiments, about 50 mg of Compound 1 is administered to a subject twice a day. In other embodiments, about 200 mg of Compound 1 is administered to a subject twice a day. In other embodiments, about 225 mg of Compound 1 is administered to a subject twice a day. In other embodiments, about 250 mg of Compound 1 is administered to a subject twice a day. In other embodiments, about 275 mg of Compound 1 is administered to a subject twice a day. In particularly preferred embodiments, about 300 mg of Compound 1 is administered to a subject twice a day. In other embodiments, about 325 mg of Compound 1 is administered to a subject twice a day. In other embodiments, about 350 mg of Compound 1 is administered to a subject twice a day. In other embodiments, about 375 mg of Compound 1 is administered to a subject twice a day. In other embodiments, about 400 mg of Compound 1 is administered to a subject twice a day.

[0340] In certain embodiments, about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, or about 500 mg of Compound 1 is administered to a subject once a day. In certain embodiments, about 50 mg of Compound 1 is administered to a subject once a day. In certain embodiments, about 75 mg of Compound 1 is administered to a subject once a day. In certain embodiments, about 100 mg of Compound 1 is administered to a subject once a day. In certain embodiments, about 125 mg of Compound 1 is administered to a subject once a day. In certain embodiments, about 150 mg of Compound 1 is administered to a subject once a day.

[0341] In certain preferred embodiments, Compound 1 is administered orally to the subject. In certain embodiments, about 50 mg of Compound 1 is administered orally to the subject twice a day. In other embodiments, about 200 mg of Compound 1 is administered orally to the subject twice a day. In other embodiments, about 250 mg of Compound 1 is administered orally to the subject twice a day. In particularly preferred embodiments, about 300 mg of Compound 1 is administered orally to the subject twice a day. In other embodiments, about 325 mg of Compound 1 is administered orally to the subject twice a day. In other embodiments, about 350 mg of Compound 1 is administered orally to the subject twice a day. In other embodiments, about 375 mg of Compound 1 is administered orally to the subject twice a day. In other embodiments, about 400 mg of Compound 1 is administered orally to the subject twice a day. In other embodiments, about 50 mg of Compound 1 is administered to the subject once a day. In other embodiments, about 75 mg of Compound 1 is administered to the subject once a day. In other embodiments, about 100 mg of Compound 1 is administered to the subject once a day. In other embodiments, about 125 mg of Compound 1 is administered to the subject once a day. In other embodiments, about 150 mg of Compound 1 is administered to the subject once daily.

[0342] In other embodiments, the IRAK4 inhibitor is PF-06650833 or BAY 1830839.

[0343] IRAK4 degraders

[0344] In certain embodiments, the method comprises administering an IRAK4 degrader. In certain embodiments, the IRAK4 degrader is KT-474.

[0345] Combination therapy

[0346] In certain embodiments of the methods disclosed herein, the method further comprises administering a BCL-2 inhibitor to the subject in combination. In certain preferred embodiments, the BCL-2 inhibitor is venetoclax. In certain embodiments, the method further comprises administering 400 mg of venetoclax daily. In certain embodiments, venetoclax is administered orally. In certain preferred embodiments, the method further comprises administering 400 mg of venetoclax orally daily.

[0347] In other embodiments, the method further comprises administering a BTK inhibitor to the subject in combination. In certain embodiments, the BTK inhibitor is ibrutinib, acalabrutinib, zabutinib, ivotinib, ONO-4059, sipetinib or HM71224. In certain embodiments, the BTK inhibitor is ibrutinib, acalabrutinib, zabutinib, ivotinib, ONO-4059, sipetinib or HM7 1224. In certain embodiments, the BTK inhibitor is acalabrutinib. In certain embodiments, the method comprises administering 200 mg of acalabrutinib daily. In certain embodiments, acalabrutinib is administered orally. In certain embodiments, the method comprises administering 200 mg of acalabrutinib orally daily. In certain preferred embodiments, the BTK inhibitor is ibrutinib. In certain embodiments, the method comprises administering 420 mg of ibrutinib daily. In other embodiments, the method comprises administering 420 mg of ibrutinib daily. In certain embodiments, ibrutinib is administered orally. In certain preferred embodiments, 420 mg of ibrutinib is administered orally daily. In other preferred embodiments, the methods comprise administering 560 mg of ibrutinib daily. In certain embodiments, the BTK inhibitor is zanubrutinib. In certain embodiments, the methods administer 160 mg of zanubrutinib twice daily. In other embodiments, the methods comprise administering 320 mg of zanubrutinib once daily. In certain embodiments, zanubrutinib is administered orally. In certain embodiments, the methods comprise administering 160 mg of zanubrutinib orally twice daily. In other embodiments, the methods comprise administering 320 mg of zanubrutinib orally once daily. In certain embodiments, the method further comprises administering one or more of the following in combination: ABT-737, BAY-1143572, 5-fluorouracil, abiraterone acetate, acetylcholine, ado-trastuzumab emtansine, afatinib, aldesleukin, alectinib, alemtuzumab, alitretinoin, aminolevulinic acid, anastrozole, anastrozole, aprepitant, arsenic trioxide, asparaginaseerwiniachrysanthemi), atezolizumab, axitinib, azacitidine, belinostat, bendamustine, benzyl isothiocyanate, bevacizumab, bexarotene, bicalutamide, bleomycin, blinatumomab, bortezomib, bosutinib, brentuximab vedotin), busulfan, cabazitaxel, capecitabine, carboplatin, carfilzomib, carmustine, ceritinib, cetuximab, chlorambucil, cisplatin, clofarabine, cobimetinib, copanlisib, crizotinib ib), cyclophosphamide, cytarabine, dabrafenib, dacarbazine, dacarbazine, dactinomycin, daratumumab, dasatinib, daunorubicin, decitabine, defibrotidesodium, degarelix, denileukin diftitox, denosumab, dexamethasone, dexrazoxane, dihydrotestosterone (DHT), dinutuximab, docetaxel, doxorubicin, elotuzumab, eltrombopag, enzalutamide, epirubicin, eribulin mesylatemesylate, erlotinib, etoposide, everolimus, exemestane, exemestane, filgrastim, fludarabine phosphate, flutamide, fulvestrant, fulvestrant, gefitinib, gemcitabine, gemtuzumab, gemtuzumab ozogamicin, glucarpidase, goserelin acetate, hydroxyurea, ibritumomab tiuxetan tiuxetan), ibrutinib, idarubicin, idelalisib, ifosfamide, imatinib, imiquimod, interferon alpha-2b, ipilimumab, irinotecan, ixabepilone, ixazomib, lanreotide, lapatinib, lenalidomide, lenvatinib, letrozole, leucovorin, leuprolide, lomustine, lomustine, mechlorethamine, megestrolacetate, melphalan, mercaptopurine, mesna, methotrexate, mitomycin C, mitoxantrone, navitoclax, necitumumab, nelarabine, netupitant, nilotinib, nilutamide, nivolumab, obinutuzumab, ofatumumab, olaparib, omacetaxinemepesuccinate, osimertinib, oxaliplatin, ozogamicin, paclitaxel, palbociclib, palifermin, pamidronate, panitumumab, panobinostat, pazopanib, pegaspargase, peginterferon alfa-2b, pembrolizumab, pemetrexed, pertuzumab, plerixafor, pomalidomide, ponatinib, pralatrexate, prednisone, procarbazine, propranolol, radium 223 dichloride, raloxifene, ramucirumab, rasburicase, regorafenib, rituximab, rolapitant, romidepsin, romiplostim, ruxolitinib, siltuximab, sipuleucel-t, sonidegib, sorafenib, sunitinib, talimogenelaherparepvec, tamoxifen, temozolomide, temsirolimus, thalidomide, thioguanine, thiotepa, tipiracil, topotecan, toremifene, toremifene, tositumomab, trabectedin, trametinib, trastuzumab In some embodiments, the second therapeutic agent is one or more of rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone.

[0348] Diseases and conditions

[0349] The methods disclosed herein relate to the treatment of many diseases and conditions; for example, the methods can be used to treat diseases and conditions associated with IRAK4. In certain embodiments, the disease or condition is cancer, preferably a hematological malignancy, such as a leukemia or lymphoma, e.g., non-Hodgkin's lymphoma. In certain embodiments, the hematological malignancy is myeloid leukemia, myeloid leukemia (e.g., acute myeloid leukemia), myelodysplastic syndrome, lymphoblastic leukemia (e.g., acute lymphoblastic leukemia), chronic lymphoblastic leukemia (CLL), small lymphocytic lymphoma (SLL), high-risk CLL, follicular lymphoma, diffuse large B-cell lymphoma (DLBCL) (e.g., DLBCL or ABC-DLBLC), mantle cell lymphoma (MCL), Waldenstrom's macroglobulinemia (WM), multiple myeloma, marginal zone lymphoma (MZL), Burkitt's lymphoma, non-Burkitt's high-grade B-cell lymphoma, extranodal marginal zone B-cell lymphoma, transformed high-grade B-cell lymphoma (HGBL), lymphoplasmacytic lymphoma (LPL), central nervous system lymphoma (CNSL) or MALT lymphoma. In certain embodiments, the hematological malignancy is myeloid leukemia. In other embodiments, the hematological malignancy is myeloid leukemia (e.g., acute myeloid leukemia). In certain embodiments, the hematological malignancy is acute myeloid leukemia (e.g., AML). In certain embodiments, AML is primary AML. In other embodiments, AML is secondary AML. In other embodiments, the hematological malignancy is myelodysplastic syndrome. In certain embodiments, myelodysplastic syndrome is high-grade. In other embodiments, myelodysplastic syndrome is low-grade. In certain embodiments, myelodysplastic syndrome is high-risk. In other embodiments, the hematological malignancy is lymphoblastic leukemia (e.g., acute lymphoblastic leukemia). In other embodiments, the hematological malignancy is chronic lymphoblastic leukemia (CLL). In certain embodiments, CLL is high-risk CLL. In other embodiments, the hematological malignancy is small lymphocytic lymphoma (SLL). In other embodiments, the hematological malignancy is follicular lymphoma. In other embodiments, the hematological malignancy is diffuse large B-cell lymphoma (DLBCL). In other embodiments, the hematological malignancy is activated B cell-like (ABC) DLBCL. In other embodiments, the hematological malignancy is germinal center B cell-like (GCB) DLBCL. In certain embodiments, DLBCL is extranodal. In certain embodiments, DLBCL is extranodal leg lymphoma, extranodal testicular lymphoma, or extranodal nonspecific (NOS) lymphoma. In other embodiments, the hematological malignancy is mantle cell lymphoma. In additional embodiments, the hematological malignancy is Waldenstrom's macroglobulinemia. In other embodiments, the hematological malignancy is multiple myeloma.In other embodiments, the hematological malignancy is marginal zone lymphoma. In other embodiments, the hematological malignancy is Burkitt's lymphoma. In other embodiments, the hematological malignancy is non-Burkitt's high-grade B-cell lymphoma. In other embodiments, the hematological malignancy is extranodal marginal zone B-cell lymphoma. In other embodiments, the hematological malignancy is transformed high-grade B-cell lymphoma (HGBL). In other embodiments, the hematological malignancy is lymphoplasmacytic lymphoma (LPL). In other embodiments, the hematological malignancy CNS lymphoma. In other embodiments, the CNS lymphoma is primary CNS lymphoma (PCNSL). In other embodiments, the hematological malignancy is MALT lymphoma. In certain embodiments, the above-mentioned hematological malignancy is relapsed or refractory. In certain embodiments, the above-mentioned hematological malignancy is resistant to treatment with a BTK inhibitor. In certain embodiments, the above-mentioned hematological malignancy is resistant to treatment with a BTK inhibitor as a monotherapy. In certain embodiments, the hematological malignancy is resistant to treatment with ibrutinib, acalabrutinib, zanubrutinib, ivotinib, ONO-4059, sepretinib, or HM7 1224. In certain preferred embodiments, the hematological malignancy is resistant to treatment with ibrutinib.

[0350] In certain embodiments, the cancer is selected from brain cancer, kidney cancer, liver cancer, stomach cancer, penis cancer, vaginal cancer, ovarian cancer, gastric cancer, breast cancer, bladder cancer, colon cancer, prostate cancer, pancreatic cancer, lung cancer, cervical cancer, epidermal cancer, prostate cancer, head and neck cancer. In certain preferred embodiments, cancer is pancreatic cancer. In other embodiments, cancer is colon cancer. In certain embodiments, cancer is a solid tumor. In various such embodiments, cancer can be recurrent or refractory. In certain embodiments, the above-mentioned cancer is resistant to treatment with BTK inhibitors. In certain embodiments, the above-mentioned cancer is resistant to treatment with BTK inhibitors as monotherapy. In certain embodiments, cancer is resistant to treatment with ibrutinib, acalabrutinib, zabutinib, ivotinib, ONO-4059, sipetinib or HM71224. In certain preferred embodiments, cancer is resistant to treatment with ibrutinib.

[0351] In other embodiments, the disease or condition is an inflammatory disease or condition. In certain embodiments, the inflammatory disease or condition is an autoimmune disease or condition. In certain embodiments, the inflammatory disease or condition is an ocular allergy, conjunctivitis, keratoconjunctivitis sicca, vernal conjunctivitis, allergic rhinitis, an autoimmune blood disorder, hemolytic anemia, aplastic anemia, pure red cell anemia, idiopathic thrombocytopenia, systemic lupus erythematosus,Rheumatoid arthritis, polychondritis, scleroderma, Wegener's granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, Stevens-Johnson syndrome, idiopathic sprue, autoimmune inflammatory bowel disease, ulcerative colitis, Crohn's disease, irritable bowel syndrome, celiac disease, periodontitis, hyaline membrane disease, kidney disease, glomerular disease, alcoholic liver disease, multiple sclerosis, endocrine eye disease, Grave's disease, sarcoidosis, alveolitis, chronic allergic pneumonitis, primary biliary cirrhosis, uveitis (anterior or posterior), Sjögren's syndrome, interstitial pulmonary fibrosis, psoriatic arthritis, systemic juvenile idiopathic arthritis, nephritis, vasculitis, diverticulitis, interstitial cystitis, glomerulonephritis, idiopathic kidney syndrome, minimal change disease, chronic granulomatous disease, endometriosis, leptospirosis nephropathy, glaucoma, retinal disease, headache, pain, complex regional pain syndrome, cardiomegaly, muscle atrophy, metabolic disorders, obesity, fetal growth retardation, hypercholesterolemia, heart disease, chronic heart failure, mesothelioma, anhidrotic urticaria dysplasia, Behcet's disease, incontinence pigmenti, Paget's disease, pancreatitis, hereditary periodic fever syndrome, asthma, acute lung injury, acute respiratory distress syndrome, eosinophilia, hypersensitivity, allergic reaction, fibrositis, gastritis, gastroenteritis, sinusitis, ocular allergy, silica-induced disease, chronic obstructive pulmonary disease (COPD), cystic fibrosis , acid-induced lung injury, pulmonary hypertension, polyneuropathy, cataracts, muscle inflammation and systemic sclerosis, inclusion body myositis, myasthenia gravis, thyroiditis, Addison's disease, lichen planus, appendicitis, atopic dermatitis, asthma, allergies, blepharitis, bronchiolitis, bronchitis, bursitis, cervicitis, cholangitis, cholecystitis, chronic transplant rejection, colitis, conjunctivitis, cystitis, dacryoadenitis, dermatitis, juvenile rheumatoid arthritis, encephalitis, endocarditis, endometritis, enteritis, enterocolitis, epicondylitis, epididymitis, fasciitis, allergic purpura, hepatitis, suppurative hidradenitis, immunoglobulin A nephropathy, interstitial lung disease, laryngitis, mastitis, meningitis, myelitis, myocarditis, myositis, nephritis, oophoritis, Orchitis, osteitis, otitis, pancreatitis, mumps, pericarditis, peritonitis, pharyngitis, urticaria, phlebitis, localized pneumonia, pneumonia, polymyositis, proctitis, prostatitis, pyelonephritis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis, tendinitis, tonsillitis, ulcerative colitis, vasculitis, vulvitis, alopecia areata, erythema multiforme, dermatitis herpetiformis, scleroderma, vitiligo, allergic vasculitis, urticaria, bullous pemphigoid, pemphigus vulgaris, pemphigus foliaceus, paraneoplastic pemphigus, acquired epidermolysis bullosa, acute or chronic gout, chronic gouty arthritis, psoriasis, psoriatic arthritis, rheumatoid arthritis, cryptopyrin-associated periodic syndrome (CAPS) and osteoarthritis. In certain preferred embodiments,Inflammatory diseases or conditions are hypercytokinemia. In certain embodiments, hypercytokinemia is caused by an infectious agent. In certain embodiments, the infectious agent is a virus. In certain preferred embodiments, the virus is a coronavirus (e.g., COVID-19). In other embodiments, the infectious agent is a bacterium. In certain embodiments, the inflammatory disease or condition is graft-versus-host disease (GVHD). In certain embodiments, GVHD is chronic graft-versus-host disease (cGVHD). In certain embodiments, GVHD is scleroderma-like GVHD, steroid-resistant GVHD, cyclosporin-resistant GVHD, GVHD, oral GVHD, reticular oral GVHD, erosive GVHD, or ulcerative oral GVHD. In certain embodiments, GVHD is scleroderma-like GVHD. In certain embodiments, GVHD is oral GVHD. In certain embodiments, GVHD is reticular oral GVHD. In certain embodiments, GVHD is erosive GVHD. In certain embodiments, GVHD is ulcerative oral GVHD. In certain embodiments, the GVHD is overlapping chronic GVHD. In certain embodiments, the GVHD is classic chronic GVHD. In certain embodiments, the GVHD is steroid-resistant GVHD. In certain embodiments, the GVHD is cyclosporine-resistant GVHD. In certain embodiments, the GVHD is refractory. In certain embodiments, the GVHD is relapsed.

[0352] In certain embodiments, the above-mentioned diseases or conditions are resistant to treatment with a BTK inhibitor alone. In certain embodiments, the above-mentioned diseases or conditions are resistant to treatment with a BTK inhibitor as a monotherapy. In certain embodiments, the disease or condition is resistant to treatment with ibrutinib, acalabrutinib, zanubrutinib, ivotinib, ONO-4059, sepretinib or HM71224. In certain preferred embodiments, the disease or condition is resistant to treatment with ibrutinib.

[0353] In certain embodiments, the disease or condition is associated with chronic anemia. In certain embodiments, the disease or condition is chronic anemia. In certain embodiments, the disease or condition is associated with transfusion dependence.

[0354] In certain embodiments, the subject is an adult.

[0355] In certain embodiments, the IRAK4 inhibitor is Compound 1; Compound 1 is orally administered once daily at a dose of about 50 mg; and the disease or condition is DLBCL. In certain embodiments, DLBCL is relapsed or refractory.

[0356] In certain embodiments, the IRAK4 inhibitor is Compound 1; Compound 1 is orally administered once daily at a dose of about 50 mg; and the disease or condition is FL. In certain embodiments, FL is relapsed or refractory.

[0357] In certain embodiments, the IRAK4 inhibitor is Compound 1; Compound 1 is orally administered once daily at a dose of about 300 mg; and the disease or disorder is WM. In certain embodiments, WM is relapsed or refractory.

[0358] In certain embodiments, the IRAK4 inhibitor is Compound 1; Compound 1 is orally administered twice daily at a dose of about 50 mg; and the disease or condition is DLBCL. In certain embodiments, DLBCL is relapsed or refractory.

[0359] In certain embodiments, the IRAK4 inhibitor is Compound 1; Compound 1 is administered orally twice daily at a dose of about 300 mg; and the disease or disorder is LPL. In certain embodiments, LPL is relapsed or refractory.

[0360] In certain embodiments, the IRAK4 inhibitor is Compound 1; Compound 1 is orally administered twice daily at a dose of about 300 mg; and the disease or condition is GCB DLBCL. In certain embodiments, GCB DLBCL is relapsed or refractory.

[0361] In certain embodiments, the IRAK4 inhibitor is Compound 1; Compound 1 is administered orally twice daily at a dose of about 400 mg; and the disease or condition is ABC DLBCL. In certain embodiments, ABC DLBCL is relapsed or refractory.

[0362] In certain embodiments, the IRAK4 inhibitor is Compound 1; Compound 1 is administered orally twice daily at a dose of about 400 mg; and the disease or condition is MZL. In certain embodiments, MZL is relapsed or refractory.

[0363] In certain embodiments, the IRAK4 inhibitor is Compound 1; Compound 1 is administered orally twice daily at a dose of about 300 mg; and the disease or condition is MZL. In certain embodiments, MZL is relapsed or refractory.

[0364] In certain embodiments, the IRAK4 inhibitor is Compound 1; Compound 1 is administered orally twice daily at a dose of about 300 mg; and the disease or condition is MALT. In certain embodiments, MALT is relapsed or refractory.

[0365] In certain embodiments, compound 1 is administered continuously (e.g., compound 1 is administered without a drug holiday). In other embodiments, compound 1 is administered intermittently (e.g., compound 1 is administered continuously interrupted by one or more drug holidays). In certain embodiments, each drug holiday lasts for a period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 days. In certain preferred embodiments, the drug holiday lasts for 7 days. In a further preferred embodiment, compound 1 is administered daily for three weeks, followed by a one-week drug holiday, optionally followed by three weeks of daily administration and a one-week drug holiday, and the cycle may be further repeated. In certain embodiments, the aforementioned dosing regimen continues, alternating dosing periods with holidays, until changes in the disease state are observed (e.g., until a complete response, a partial response, or unacceptable toxicity is observed). Methods for treating certain diseases and conditions with compound 1 are disclosed in PCT / U20S21 / 030192, the contents of which are incorporated herein by reference in their entirety.

[0366] Previous treatment

[0367] The methods disclosed herein can be used as a first-line therapy, or can be applied to patients who cannot achieve a partial or complete response using one or more previous anticancer therapies or anti-inflammatory therapies. In certain embodiments, the subject has previously received at least one anticancer therapy. In certain embodiments, the patient has previously received one anticancer therapy. In other embodiments, the patient has previously received two anticancer therapies. In other embodiments, the patient has previously received three anticancer therapies. In other embodiments, the patient has previously received four anticancer therapies. In other embodiments, the patient has previously received five anticancer therapies. In certain embodiments, at least one anticancer therapy is selected from anti-CD20 antibodies, nitrogen mustards, steroids, purine analogs, DNA topoisomerase inhibitors, DNA intercalators, tubulin inhibitors, BCL-2 inhibitors, proteasome inhibitors, toll-like receptor inhibitors, kinase inhibitors, SRC kinase inhibitors, PI3K kinase inhibitors, BTK inhibitors, glutaminase inhibitors, PD-1 inhibitors, PD-L1 inhibitors and methylating agents; or a combination thereof. In certain embodiments, the anticancer therapy is selected from ibrutinib, rituximab, bendamustine, bortezomib, dexamethasone, chlorambucil, cladribine, cyclophosphamide, doxorubicin, vincristine, venetoclax, ifosfamide, prednisone, oprozomib, ixazomib, acalabrutinib, zanubrutinib, IMO-08400, idelalisib, erbulisel, CB-839, fludarabine, and thalidomide; or a combination thereof. In certain embodiments, the anticancer therapy is ibrutinib. In certain embodiments, the anticancer therapy is ibrutinib and rituximab. In certain embodiments, the anticancer therapy is bendamustine. In certain embodiments, the anticancer therapy is bendamustine and rituximab. In certain embodiments, the anticancer therapy is bortezomib. In certain embodiments, the anticancer therapy is bortezomib and dexamethasone. In certain embodiments, anticancer therapy is bortezomib and rituximab. In certain embodiments, anticancer therapy is bortezomib, rituximab and dexamethasone. In certain embodiments, chlorambucil. In certain embodiments, anticancer therapy is cladribine. In certain embodiments, anticancer therapy is cladribine and rituximab. In certain embodiments, anticancer therapy is cyclophosphamide, doxorubicin, vincristine, prednisone and rituximab (ie, CHOP-R). In certain embodiments, anticancer therapy is cyclophosphamide, prednisone and rituximab (ie, CPR). In certain embodiments, anticancer therapy is fludarabine. In certain embodiments, anticancer therapy is fludarabine and rituximab. In certain embodiments, anticancer therapy is fludarabine, cyclophosphamide and rituximab. In certain preferred embodiments, anticancer therapy is rituximab. In certain preferred embodiments, anticancer therapy comprises rituximab.In certain embodiments, anticancer therapy is rituximab, cyclophosphamide and dexamethasone (ie, RCD). In certain embodiments, anticancer therapy is thalidomide. In certain embodiments, anticancer therapy is thalidomide and rituximab. In certain embodiments, anticancer therapy is venetoclax. In certain embodiments, anticancer therapy is cyclophosphamide, bortezomib and dexamethasone (ie, R-CyBorD). In certain embodiments, anticancer therapy is a hypomethylating agent. In certain embodiments, the subject has previously received at least 6 cycles of hypomethylating agents. In certain embodiments, anticancer therapy is any of the aforementioned combinations, for example, the subject may first receive rituximab, and then receive a combination of rituximab, cyclophosphamide and dexamethasone (ie, RCD) later.

[0368] In certain embodiments, the subject has previously received at least one anti-inflammatory therapy. In certain embodiments, the patient has previously received one anti-inflammatory therapy. In other embodiments, the patient has previously received two anti-inflammatory therapies. In other embodiments, the patient has previously received three anti-inflammatory therapies. In other embodiments, the patient has previously received four anti-inflammatory therapies. In certain embodiments, the anti-inflammatory agent is a steroid (e.g., a corticosteroid). In certain embodiments, the anti-inflammatory therapy is hydrocortisone, cortisone, ethamethasone (ethamethasoneb), prednisone, prednisolone, triamcinolone, dexamethasone or fludrocortisone; or a combination thereof.

[0369] The subject may also have received or is preparing to undergo other non-chemotherapeutic treatments, such as surgery, radiotherapy, or bone marrow transplantation. In certain embodiments, the subject has previously received etoposide chemomobilization therapy. In certain embodiments, the subject has previously received bone marrow transplantation. In certain embodiments, the subject has previously received stem cell transplantation. In certain embodiments, the subject has previously received autologous stem cell transplantation. In certain embodiments, the subject has previously received allogeneic stem cell transplantation. In certain embodiments, the subject has previously received hematopoietic cell transplantation. In certain embodiments, the subject has previously received carmustine, etoposide, cytarabine, and melphalan (i.e., BEAM conditioning). In certain embodiments, the subject has previously received re-induction therapy.

[0370] The subject may also have previously shown favorable results to a previous therapy, only to require additional treatment later. In certain embodiments, the subject has previously achieved a partial response. In certain embodiments, the subject has previously achieved a good partial response. In certain embodiments, the subject has previously achieved a complete response. In certain embodiments, the cancer is recurrent. In certain embodiments, the cancer is refractory.

[0371] The subject may also have one or more pre-existing or developed gene mutations that make the subject's cancer more or less resistant to therapy. In certain embodiments, the subject has a mutation in RICTOR. In certain embodiments, there is an N1065S mutation in RICTOR. In certain preferred embodiments, the subject has a mutation in MYD88. In certain even more preferred embodiments, the subject has an L265P mutation in MYD88. In certain embodiments, the subject has a mutation in TET2. In certain embodiments, the subject does not have a mutation in CXCR4. In other embodiments, the subject has a mutation in CXCR4. In certain embodiments, the subject shows early progression. In certain embodiments, the subject has not previously received a BTK inhibitor.

[0372] In certain embodiments, after the administration of the compound, the subject achieves a partial response. In certain embodiments, after the administration of the compound, the subject achieves a good partial response. In other embodiments, after the administration of the compound, the subject achieves a complete response. In certain embodiments, the subject achieves a partial response within 7 days of receiving the compound. In certain embodiments, the subject achieves a good partial response within 7 days of receiving the compound. In certain embodiments, the subject achieves a complete response within 7 days of receiving the compound. In certain embodiments, the subject's tumor volume is reduced by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90% or about 95%. In certain embodiments, the subject's tumor volume is reduced by 5%. In certain embodiments, the subject's tumor volume is reduced by 10%. In certain embodiments, the subject's tumor volume is reduced by 15%. In certain embodiments, the subject's tumor volume is reduced by 20%. In certain embodiments, the tumor volume of the subject is reduced by 25%. In certain embodiments, the tumor volume of the subject is reduced by 30%. In certain embodiments, the tumor volume of the subject is reduced by 35%. In certain embodiments, the tumor volume of the subject is reduced by 40%. In certain embodiments, the tumor volume of the subject is reduced by 45%. In certain embodiments, the tumor volume of the subject is reduced by 50%. In certain embodiments, the tumor volume of the subject is reduced by 55%. In certain embodiments, the tumor volume of the subject is reduced by 60%. In certain embodiments, the tumor volume of the subject is reduced by 65%. In certain embodiments, the tumor volume of the subject is reduced by 70%. In certain embodiments, the tumor volume of the subject is reduced by 80%. In certain embodiments, the tumor volume of the subject is reduced by 85%. In certain embodiments, the tumor volume of the subject is reduced by 90%. In certain embodiments, the tumor volume of the subject is reduced by 95%.

[0373] Methods for immunohistochemical staining

[0374] In certain embodiments of the methods disclosed herein, the expression level of NF-κB p-p50 in the sample can be determined by immunohistochemical staining. Methods for performing immunohistochemical staining are generally known to those skilled in the art. In short, the tissue sample is contacted with an antibody specific for NF-κB p-p50 or NF-κB p-p65. After the incubation period, the tissue sample is contacted with a secondary antibody. The secondary antibody recognizes and binds to the first antibody. The secondary antibody may contain a conjugation activity (e.g., an enzymatic activity) for detecting the presence of the secondary antibody, and thereby detecting the presence of the first antibody, and then detecting the presence of NF-κB p-p50 or NF-κB p-p65. Exemplary conjugation activities can be any activity known to those skilled in the art that can be used to produce a detectable immunohistochemical signal. Enzyme conjugates suitable for the secondary antibody include, for example, horseradish peroxidase (HRP), alkaline phosphatase, glucose oxidase, and β-galactosidase; fluorescent probes, radioisotopes, chemiluminescent compounds, bioluminescent compounds, or combinations thereof are also contemplated.

[0375] In certain embodiments, the NF-κB p-p50 or NF-κB p-p65 specific antibody is a commercially available NF-κB p-p50 antibody. In certain embodiments, the NF-κB p-p50 or NF-κB p-p65 specific antibody is a polyclonal antibody. In certain embodiments, the NF-κB p-p50 or NF-κB p-p65 specific antibody is a monoclonal antibody. In certain embodiments, the NF-κB p-p50 or NF-κB p-p65 specific antibody is a rabbit antibody. In certain embodiments, the NF-κB p-p50 specific antibody is a phosphorylated p50 NF-κB (Ser337) (sc-271908) Ab from Santa Cruz Biotechnology. In certain embodiments, the NF-κB p-p65 specific antibody is a phosphorylated p65 NF-κB (Ser536) (ab86299) Ab from Abcam. In certain embodiments, the NF-κB p-p65 specific antibody is the phosphorylated p65 NF-κB (Ser276) (ab194726) Ab from Abeam.

[0376] In certain embodiments, the secondary antibody is commercially available.In certain embodiments, the secondary antibody is a peroxidase-labeled polymer conjugated to goat anti-rabbit immunoglobulin, such as those included in the EnVision+ System-HRP kit (DAKO, Carpinteria, CA).

[0377] In the methods of the present disclosure, after determining the expression level of NF-κB p-p50 or NF-κB p-p65 in a tissue sample, the level is compared with the expression level of NF-κB p-p50 in a reference sample. In certain embodiments, the reference sample has the same or comparable tissue type as the tissue sample, but is known to have a normal expression level of NF-κB p-p50 or NF-κB p-p65 or does not express NF-κB p-p50 or NF-κB p-p65. In certain embodiments, the reference sample is a normal or non-diseased tissue of the same tissue type as the tissue sample, but is taken from an individual or group of individuals known to express a normal expression level of NF-κB p-p50 or NF-κB p-p65 or does not express NF-κB p-p50 or NF-κB p-p65. In certain embodiments, the reference sample is a normal or non-diseased tissue of the same or comparable tissue type as a tissue sample taken from the same individual as the tissue sample. In certain embodiments, the reference sample comprises a normal or non-diseased cell subpopulation within the tissue sample. In certain embodiments, the reference sample is a plurality of cells or tissues that do not exhibit a phenotype of elevated expression levels of NF-κB p-p50 or NF-κB p-p65.

[0378] When the expression level of NF-κB p-p50 or NF-κB p-p65 in the tissue sample is higher than the expression level of NF-κB p-p50 or NF-κB p-p65 in the reference sample, an increased expression level will be detected. Positive expression of NF-κB p-p50 or NF-κB p-p65 can be defined as positive staining of the cytoplasm and / or nucleus of more than 50% of the cancer cells.

[0379] In the immunohistochemical staining method of the present disclosure, a biological sample is obtained. The biological sample can be any tissue specimen or any cell collection from a tissue. The biological sample can be from any animal or human. In certain embodiments, the biological sample is from a human. In other embodiments, the biological sample is from an animal.

[0380] In the immunohistochemical staining method of the present disclosure, a biological sample is contacted with a first antibody specific for NF-κB p-p50 or NF-κB p-p65 to produce a biological sample contacted with the first antibody. The first antibody is specific for NF-κB p-p50, which means that the antibody selectively binds to NF-κB p-p50 or NF-κB p-p65. In certain embodiments, the first antibody is a polyclonal antibody. In certain embodiments, the first antibody is a monoclonal antibody. In certain embodiments, the first antibody is a rabbit polyclonal antibody. In certain embodiments, the first antibody is a rabbit monoclonal antibody.

[0381] In the immunohistochemical staining method of the present disclosure, a biological sample contacted with a first antibody is contacted with a second antibody that is specific for the first antibody, wherein the second antibody also has conjugation activity. The second antibody must selectively bind to the first antibody. The second antibody may be from the same species as the first antibody, or from a species different from the first antibody. The second antibody may be a polyclonal antibody or a monoclonal antibody.

[0382] The second antibody also has conjugation activity, which can be an enzymatic activity. In certain embodiments, the enzymatic activity is an intrinsic activity of the second antibody. In other embodiments, the enzymatic activity of the second antibody is provided by an enzyme conjugated to the antibody.

[0383] In certain embodiments, the enzymatic activity of the second antibody is peroxidase activity. In other embodiments, the enzymatic activity of the second antibody is alkaline phosphatase activity. Exemplary conjugated enzymatic activity can be any enzymatic activity known to those skilled in the art that can be used to produce detectable immunohistochemical signals, including, for example, horseradish peroxidase (HRP), alkaline phosphatase, glucose oxidase and beta-galactosidase. Other immunohistochemical signals are also contemplated, including, for example, fluorescent probes, radioisotopes, chemiluminescent compounds, bioluminescent compounds or combinations thereof.

[0384] In the immunohistochemical staining method disclosed herein, the product after the biological sample contacted with the first antibody is contacted with the second antibody is a biological sample bound to the first antibody, and wherein the second antibody is bound to the first antibody. In the method disclosed herein, this product is contacted with a chromogenic substrate for the enzymatic activity of the second antibody.

[0385] The chromogenic substrate for the enzymatic activity of the secondary antibody is a compound that changes color after reacting with the enzymatic activity of the secondary antibody. In certain embodiments, the chromogenic substrate is diaminobenzidine (DAB). In other embodiments, the chromogenic substrate is 3-amino-9-ethylcarbazole (AEC). In other embodiments, the chromogenic substrate is 5-bromo-4-chloro-3-indolyl phosphate / tetranitro blue tetrazolium (BCIP / TNBT). In other embodiments, the chromogenic substrate is naphthol AS-MX phosphate + Fast Blue BB.

[0386] After treating the sample with the chromogenic substrate, the product is counterstained for a period of time. Any counterstain sufficient to contrast the color of the chromogenic substrate can be used. Many different counterstains are known to those skilled in the art, including, for example, methyl green and hematoxylin.

[0387] In certain embodiments, the product is then counterstained for up to 1 minute. In certain embodiments, the product is counterstained for up to 10 seconds.

[0388] In certain embodiments, the counterstain is hematoxylin. Methods of using hematoxylin are known to those skilled in the art. See, for example, Godwin Avwioro, Histochemical uses of Haematoxylin—A Review, JPCS Vol. 1, April-June 2011, 24-34. The concentration of hematoxylin is generally in the range of about 1 g / L to about 2 g / L.

[0389] Pharmaceutical composition

[0390] The compositions and methods of the present invention can be used to treat individuals in need. In certain embodiments, the individual is a mammal, such as a human or non-human mammal. When applied to an animal such as a human, the composition or compound is preferably applied in the form of a pharmaceutical composition, which comprises, for example, a compound of the present invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art, and include, for example, aqueous solutions, such as water or physiologically buffered saline or other solvents or vehicles, such as glycols, glycerol, oils such as olive oil or injectable organic esters. In a preferred embodiment, when such pharmaceutical compositions are applied to humans, particularly for invasive routes of administration (i.e., such as injections or implants that circumvent transport or diffusion through an epithelial barrier), the aqueous solution is pyrogen-free or substantially pyrogen-free. Excipients can be selected, for example, to achieve delayed release of an agent or selectively target one or more cells, tissues or organs. Pharmaceutical compositions can be in dosage unit form, such as tablets, capsules (including dispersed capsules and gelatin capsules), granules, lyophilized agents for reconstitution, powders, solutions, syrups, suppositories, injections, etc. The composition may also be presented in a transdermal delivery system, such as a skin patch. The composition may also be presented in a solution suitable for topical administration, such as a lotion, cream or ointment.

[0391] A pharmaceutically acceptable carrier may contain a physiologically acceptable agent, which, for example, acts to stabilize a compound (such as a compound of the present invention), increase its solubility, or increase its absorption. Such physiologically acceptable agents include, for example, carbohydrates such as glucose, sucrose, or dextran; antioxidants such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins or other stabilizers or excipients. The selection of a pharmaceutically acceptable carrier (including a physiologically acceptable agent) depends, for example, on the route of administration of the composition. The formulation or pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) may also be a liposome or other polymer matrix, into which, for example, a compound of the present invention may be incorporated. For example, liposomes comprising phospholipids or other lipids are non-toxic, physiologically acceptable, and metabolizable carriers that are relatively simple to prepare and administer.

[0392] The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0393] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethyl cellulose and cellulose acetate; (4) tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and and soybean oil; (10) glycols such as propylene glycol; (11) polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solution; and (21) other non-toxic compatible substances used in pharmaceutical formulations.

[0394] The pharmaceutical composition (preparation) can be administered to a subject by any of a variety of routes of administration, including, for example, oral administration (e.g., an infusion in water or non-aqueous solutions or suspensions for application to the tongue, tablets, capsules (including dispersible capsules and gelatin capsules), boluses, powders, granules, pastes); absorption through the oral mucosa (e.g., sublingually); subcutaneously; transdermally (e.g., as a patch applied to the skin); and topically (e.g., as a cream, ointment or spray applied to the skin). The compound can also be formulated for inhalation. In certain embodiments, the compound can simply be dissolved or suspended in sterile water. Details of appropriate routes of administration and compositions useful therefor may be found, for example, in U.S. Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970, and 4,172,896, and patents cited therein.

[0395] The formulation can be conveniently present in unit dosage form and can be prepared by any method well known in the pharmaceutical field. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary according to the host being treated, the specific mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be the amount of the compound that produces the therapeutic effect. In general, in one hundred parts, this amount will be in the range of about 1% to about 99% active ingredient, preferably about 5% to about 70%, and most preferably about 10% to about 30%.

[0396] Methods for preparing these formulations or compositions include the step of bringing into association the active compound (such as a compound of the present invention) with a carrier and optionally one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0397] Formulations of the invention suitable for oral administration may be in the form of capsules (including dispersible capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored base, usually sucrose and acacia or tragacanth), lyophilizates, powders, granules, or as a solution or suspension in an aqueous liquid or a non-aqueous liquid, or as an oil-in-water or water-in-oil emulsion, or as an elixir or syrup, or as a pastille (using an inert base such as gelatin and glycerin, or sucrose and acacia), and / or as a mouthwash, etc., each containing a predetermined amount of the compound of the invention as the active ingredient. The composition or compound may also be administered as a bolus, electuary, or paste.

[0398] To prepare solid dosage forms for oral administration (capsules (including dispersible capsules and gelatin capsules), tablets, pills, dragees, powders, granules, etc.), the active ingredient is mixed with one or more pharmaceutically acceptable carriers (such as sodium citrate or dicalcium phosphate) and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and / or silicic acid; (2) binders, such as carboxymethyl cellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or gum arabic; (3) humectants, such as glycerol. ; (4) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain silicates and sodium carbonate; (5) solution retardants, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compositions; (7) wetting agents, such as cetyl alcohol and glyceryl monostearate; (8) absorbents, such as kaolin and bentonite clays; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate and mixtures thereof; (10) complexing agents, such as modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including dispersible capsules and gelatin capsules), tablets and pills, the pharmaceutical composition may also contain a buffer. Similar types of solid compositions may also be used as fillers in soft-filled and hard-filled gelatin capsules using excipients such as lactose or lactose and high molecular weight polyethylene glycols.

[0399] Tablets can be prepared by compression or molding, optionally containing one or more auxiliary ingredients. Compressed tablets can be prepared using a binder (e.g., gelatin or hydroxypropyl methylcellulose), a lubricant, an inert diluent, a preservative, a disintegrant (e.g., sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), a surfactant or a dispersant. Molded tablets can be prepared by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0400] Tablets and other solid dosage forms of pharmaceutical compositions such as dragees, capsules (including dispersible capsules and gelatin capsules), pills and granules may be optionally scored or prepared with coatings and shells, such as enteric coatings or other coatings well known in the field of pharmaceutical formulation. Hydroxypropyl methylcellulose, other polymer matrices, liposomes and / or microspheres in different proportions, for example, for providing the desired release characteristics, may also be formulated to provide slow release or controlled release of the active ingredient contained therein. It may be sterilized by, for example, filtering through a bacteria-retaining filter or by incorporating a sterilizing agent in the form of a sterile solid composition, which may be dissolved in sterile water or some other sterile injectable medium immediately before use. These compositions may also optionally contain an opacifier and may have a composition that releases the active ingredient only or preferentially in a certain part of the gastrointestinal tract, optionally in a delayed manner. Examples of embedding compositions that may be used include polymeric substances and waxes. The active ingredient may also be in microencapsulated form, with one or more of the above-mentioned excipients, where appropriate.

[0401] Liquid dosage forms that can be used for oral administration include pharmaceutically acceptable emulsions, lyophilized agents for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage form may contain an inert diluent commonly used in the art, such as, for example, water or other solvents, cyclodextrins and derivatives thereof, solubilizers and emulsifiers such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (specifically cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan and mixtures thereof.

[0402] Besides inert diluents, the oral compositions may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0403] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0404] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier and any preservatives, buffers or propellants that may be required.

[0405] Ointments, pastes, creams and gels may contain, in addition to the active compounds, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0406] In addition to the active compound, powders and sprays may contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate and polyamide powder, or mixtures of these substances. Sprays may additionally contain customary propellants, such as chlorofluorocarbons and volatile unsubstituted hydrocarbons (such as butane and propane).

[0407] Transdermal patches have the additional advantage of providing controlled delivery of the compounds of the present invention to health. Such dosage forms can also be prepared by dissolving or dispersing the active compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound through the skin. The rate of this flux can be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or a gel.

[0408] As used herein, the phrases "parenteral administration" and "administered parenterally" mean modes of administration other than enteral administration and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration comprise a combination of one or more active compounds with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be reconstituted into sterile injectable solutions or dispersions immediately prior to use, which combination may contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0409] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters (such as ethyl oleate). Appropriate fluidity can be maintained, for example, by the use of coating materials (such as lecithin), by maintaining the desired particle size in the case of dispersions, and by the use of surfactants.

[0410] These compositions may also contain adjuvants, such as preservatives, wetting agents, emulsifiers and dispersants. Various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, etc., may be included to ensure that the effects of microorganisms are prevented. It may also be desirable to include isotonic agents, such as sugar, sodium chloride, etc., in the composition. In addition, extended absorption of injectable drug forms may be achieved by including agents that delay absorption, such as aluminum monostearate and gelatin.

[0411] In some cases, in order to prolong the effect of the drug, it is desirable to slow down the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of a crystalline or amorphous material with poor water solubility. The absorption rate of the drug depends on its dissolution rate, which in turn can depend on crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form can be accomplished by dissolving or suspending the drug in an oil vehicle.

[0412] Injectable depot forms are prepared by forming a microencapsulated matrix of the subject compound in a biodegradable polymer such as polylactide-polyglycolide. Depending on the ratio of the drug to the polymer and the properties of the specific polymer used, the drug release rate can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injection formulations can also be prepared by embedding the drug in a liposome or microemulsion compatible with body tissues.

[0413] For use in the methods of the invention, the active compound may be provided per se or as a pharmaceutical composition containing, for example, 0.1% to 99.5% (more preferably 0.5% to 90%) of the active ingredient in combination with a pharmaceutically acceptable carrier.

[0414] The method of introduction can also be provided by a rechargeable or biodegradable device. Regarding the controlled delivery of drugs (including protein biopharmaceuticals), various slow-release polymer devices have been developed and tested in vivo in recent years. A variety of biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, can be used to form implants to continuously release compounds at specific target sites.

[0415] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0416] The selected dosage level will depend upon a variety of factors including the activity of the specific compound or combination of compounds employed, or the esters, salts or amides thereof, the route of administration, the time of administration, the rate of excretion of the specific compound employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the specific compound employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0417] A physician or veterinarian with ordinary skills in the art can easily determine and prescribe the desired pharmaceutical composition for a therapeutically effective amount. For example, a physician or veterinarian can start the pharmaceutical composition or compound dosage at a level lower than the level required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. "Therapeutically effective amount" means the concentration of the compound sufficient to cause the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the subject's weight, sex, age and medical history. Other factors affecting the effective amount may include, but are not limited to, the severity of the patient's condition, the condition being treated, the stability of the compound, and, if desired, another type of therapeutic agent administered with the compound of the present invention. A larger total dose can be delivered by multiple administrations of the agent. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13th edition, 1814-1882, incorporated herein by reference).

[0418] In general, a suitable daily dose of the active compound used in the compositions and methods of the invention will be that amount of the compound which is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend on the factors described above.

[0419] If desired, the effective daily dose of the active compound may be administered as one, two, three, four, five, six or more sub-doses, divided at appropriate intervals throughout the day, optionally in unit dosage form. In certain embodiments of the invention, the active compound may be administered twice or three times a day. In a preferred embodiment, the active compound will be administered once a day.

[0420] Patients receiving such treatment are any animal in need thereof, including primates, particularly humans; as well as other mammals, such as horses, cattle, pigs, sheep, cats and dogs; poultry; and pets in general.

[0421] In certain embodiments, the compounds of the invention may be used alone or in combination with another type of therapeutic agent.

[0422] The disclosure includes the use of pharmaceutically acceptable salts of the compounds of the present invention in the compositions and methods of the present invention. In certain embodiments, the salts considered by the present invention include but are not limited to alkyl, dialkyl, trialkyl or tetraalkyl ammonium salts. In certain embodiments, the salts considered by the present invention include but are not limited to L-arginine, benethamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucosamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine and zinc salts. In certain embodiments, the salts considered by the present invention include but are not limited to Na, Ca, K, Mg, Zn or other metal salts. In certain embodiments, the salts contemplated by the present invention include, but are not limited to, 1-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxy-ethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, l-ascorbic acid, l-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, capric acid / decanoic acid, caproic acid / hexanoic acid, caprylic acid / octanoic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, acid), dodecyl sulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d-glucoheptonic acid, d-gluconic acid, d-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, l-malic acid, malonic acid, phenylglycolic acid, methanesulfonic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, niacin, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, propionic acid, l-pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, l-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenate.

[0423] Pharmaceutically acceptable acid addition salts may also exist in the form of various solvates, such as solvates with water, methanol, ethanol, dimethylformamide, etc. Mixtures of such solvates may also be prepared. The source of such solvates may be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or insoluble in such solvents.

[0424] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0425] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and (3) metal chelators, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0426] definition

[0427] Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings commonly understood by those of ordinary skill in the art. In general, the nomenclature and techniques used in conjunction with chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics, and protein and nucleic acid chemistry described herein are those well-known and commonly used in the art.

[0428] Unless otherwise indicated, the methods and techniques of the present disclosure are generally performed according to conventional methods well known in the art and as described in various general and more specific references cited and discussed throughout this specification. See, for example, "Principles of Neural Science", McGraw-Hill Medical, New York, NY (2000); Motulsky, "Intui tive Biostatistics", Oxford University Press, Inc. (1995); Lodish et al., "Molecular Cell Biology, 4th Edition", WH Freeman & Co., New York (2000); Griffiths et al., "Introduction to Genetic Analysis, 7th Edition", WH Freeman & Co., NY (1999); and Gilbert et al., "Developmental Biology, 6th Edition", Sinauer Associates, Inc., Sunderland, MA (2000).

[0429] Unless otherwise defined herein, chemical terms used herein are used according to conventional usage in the art, as exemplified by “The McGraw-Hill Dictionary of Chemical Terms”, Parker S., ed., McGraw-Hill, San Francisco, CA (1985).

[0430] All of the above and any other publications, patents, and published patent applications mentioned in this patent application are expressly incorporated herein by reference. In the event of conflict, the present specification, including its specific definitions, will control.

[0431] The term "agent" as used herein refers to a compound (such as an organic or inorganic compound, a mixture of compounds), a biomacromolecule (such as a nucleic acid, an antibody, including portions thereof and humanized, chimeric and human antibodies and monoclonal antibodies, a protein or portions thereof, such as peptides, lipids, carbohydrates) or an extract made from biological materials such as bacteria, plants, fungi or animal (particularly mammalian) cells or tissues. Agents include, for example, agents of known structure and agents of unknown structure. The ability of such agents to inhibit AR or promote AR degradation may make them suitable as "therapeutic agents" in the methods and compositions of the present disclosure.

[0432] "Patient", "subject" or "individual" are used interchangeably and refer to human or non-human animals. These terms include mammals, such as humans, primates, livestock animals (including cattle, pigs, etc.), companion animals (e.g., dogs, cats, etc.), and rodents (e.g., mice and rats).

[0433] "Treating" a condition or patient means taking steps to obtain a beneficial or desired result, including a clinical result. Beneficial or desired clinical results may include, but are not limited to, alleviation or improvement of one or more symptoms or conditions, reduction in the extent of the disease, stabilization of the disease state (i.e., no worsening), prevention of disease spread, delay or slowing of disease progression, improvement or alleviation of the disease state, and remission (whether partial or total), whether detectable or undetectable. "Treatment" may also mean prolonged survival compared to survival expected when not receiving treatment.

[0434] The term "prevention" is art-recognized and when used with respect to a condition such as a local recurrence (e.g., pain), a disease such as cancer, a symptom such as heart failure, or any other medical condition, is well known in the art and includes administering a composition that reduces the frequency of symptoms of the medical condition in a subject, or delays the onset of the same, relative to a subject not receiving the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable cancerous growths in a patient population receiving prophylactic treatment relative to an untreated control population, and / or delaying the appearance of detectable cancerous growths in a treated population relative to an untreated control population, e.g., by a statistically and / or clinically significant amount.

[0435] "Administering / administration" a substance, compound or agent to a subject can be implemented using one of the various methods known to those skilled in the art. For example, a compound or agent can be administered by intravenous, intraarterial, intradermal, intramuscular, intraperitoneal, subcutaneous, ocular, sublingual, oral (by ingestion), intranasal (by inhalation), intraspinal, intracerebral, and transdermal (by absorption, for example, through a skin tube). A compound or agent can also be suitably introduced by a rechargeable or biodegradable polymer device or other device (such as a patch and a pump) or formulation that provides extended, slow or controlled release of the compound or agent. Administration can also be performed, for example, once, multiple times, and / or over one or more extended time periods.

[0436] The appropriate method of administering a substance, compound or agent to a subject will also depend on, for example, the age and / or physical condition of the subject and the chemical and biological properties (e.g., solubility, digestibility, bioavailability, stability and toxicity) of the compound or agent. In certain embodiments, the compound or agent is orally administered to a subject, for example, by ingestion. In certain embodiments, the orally administered compound or agent is in a formulation that extends release or slow release, or is administered using a device for this slow or extended release.

[0437] As used herein, the phrase "combined administration" refers to any form of administration of two or more different therapeutic agents such that a second agent is administered while the previously administered therapeutic agent is still effective in vivo (e.g., both agents are effective in the patient simultaneously, which may include a synergistic effect of the two agents). For example, different therapeutic compounds may be administered simultaneously or sequentially in the same formulation or in separate formulations. Thus, an individual receiving such treatment may benefit from the combined effects of the different therapeutic agents.

[0438] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or agent is an amount of the drug or agent that will have the intended therapeutic effect when administered to a subject. The full therapeutic effect does not necessarily occur by administering a single dose, but may only occur after a series of doses are administered. Thus, a therapeutically effective amount may be administered in one or more administrations. The precise effective amount required for a subject will depend, for example, on the subject's size, health, and age, and the nature and extent of the condition being treated (such as cancer or MDS). A skilled person can readily determine the effective amount for a given situation by routine experimentation.

[0439] As used herein, the term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where the event or circumstance does not occur. For example, "optionally substituted alkyl" means that the alkyl group may be substituted as well as instances where the alkyl group is not substituted.

[0440] It is understood that substituents and substitution patterns on the compounds of the invention can be selected by one of ordinary skill in the art to yield chemically stable compounds that can be readily synthesized from readily available starting materials by techniques known in the art and those described below. If a substituent is itself substituted with more than one group, it is understood that these multiple groups may be on the same carbon or on different carbons, as long as a stable structure results.

[0441] As used herein, the term "optionally substituted" refers to the replacement of one to six hydrogen groups in a given structure with the group of a specified substituent, including but not limited to: hydroxy, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silyl, acyl, acyloxy, aryl, cycloalkyl, heterocyclyl, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OCO-CH2-O-alkyl, -OP(O)(O-alkyl)2 or -CH2-OP(O)(O-alkyl)2. Preferably, "optionally substituted" refers to the replacement of one to four hydrogen groups in a given structure with the substituents mentioned above. More preferably, one to three hydrogen groups are replaced by the substituents mentioned above. It should be understood that the substituents may be further substituted.

[0442] As used herein, the term "alkyl" refers to a saturated aliphatic group, including but not limited to C1-C 10 Straight chain alkyl group or C1-C 10Branched alkyl groups. Preferably, "alkyl" groups refer to C1-C6 straight chain alkyl groups or C1-C6 branched chain alkyl groups. Most preferably, "alkyl" groups refer to C1-C4 straight chain alkyl groups or C1-C4 branched chain alkyl groups. The example of "alkyl" includes but is not limited to methyl, ethyl, 1-propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, 1-pentyl, 2-pentyl, 3-pentyl, neopentyl, 1-hexyl, 2-hexyl, 3-hexyl, 1-heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 1-octyl, 2-octyl, 3-octyl or 4-octyl etc. "alkyl" groups can be optionally substituted.

[0443] The term "acyl" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

[0444] The term "acylamino" is art-recognized and refers to an amino group substituted with an acyl group, and may be represented, for example, by the formula hydrocarbyl C(O)NH-.

[0445] The term "acyloxy" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-.

[0446] The term "alkoxy" refers to an alkyl group attached to an oxygen. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy, and the like.

[0447] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group, and may be represented by the general formula alkyl-O-alkyl.

[0448] The term "alkyl" refers to a saturated aliphatic group, including straight chain alkyl, branched chain alkyl, cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl and cycloalkyl substituted alkyl. In preferred embodiments, a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., for a straight chain alkyl having 30 carbon atoms or less, for a straight chain alkyl having 30 carbon atoms or less, for a branched ... 1-30 , for the branched chain C 3-30 ), and more preferably 20 or less.

[0449] In addition, the term "alkyl" as used throughout the specification, examples and claims is intended to include unsubstituted and substituted alkyl groups, the latter referring to alkyl moieties having substituents replacing hydrogen on one or more carbon atoms of the hydrocarbon backbone, including halogenated alkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl and the like.

[0450] When used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, the term "C x-y ” or “C x -C y" is meant to include groups containing from x to y carbons in the chain. C0 alkyl represents hydrogen, where the group is in a terminal position, or a bond if internal. For example, C 1-6 Alkyl groups contain 1 to 6 carbon atoms in the chain.

[0451] As used herein, the term "alkylamino" refers to an amino group substituted with at least one alkyl group.

[0452] As used herein, the term "alkylthio" refers to a thiol group substituted with an alkyl group, and may be represented by the general formula alkylS-.

[0453] As used herein, the term "amide" refers to a group

[0454]

[0455] Where R 9 and R 10 Each independently represents hydrogen or a hydrocarbon group, or R 9 and R 10 Together with the N atom to which it is attached it forms a heterocyclic ring having 4 to 8 atoms in the ring structure.

[0456] The terms "amine" and "amino" are art-recognized and refer to unsubstituted and substituted amines and salts thereof, for example, a moiety represented by the formula

[0457]

[0458] Where R 9 , R 10 and R 10 ' each independently represents hydrogen or a hydrocarbon group, or R 9 and R 10 Together with the N atom to which it is attached it forms a heterocyclic ring having 4 to 8 atoms in the ring structure.

[0459] As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group.

[0460] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group.

[0461] As used herein, the term "aryl" includes substituted or unsubstituted monocyclic aromatic groups, wherein each atom of the ring is carbon. Preferably, the ring is a 5-7-membered ring, more preferably a 6-membered ring. The term "aryl" also includes a polycyclic system with two or more rings, wherein two or more carbons are common to two adjacent rings, wherein at least one ring is aromatic, for example, the other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclic. Aryl includes benzene, naphthalene, phenanthrene, phenol, aniline, etc.

[0462] The term "carbamate" is art-recognized and refers to the group

[0463]

[0464] Where R 9 and R 10 independently represents hydrogen or a hydrocarbyl group.

[0465] As used herein, the term "carbocyclylalkyl" refers to an alkyl group substituted with a carbocyclyl group.

[0466] The term "carbocycle" includes 5-7 membered monocycles and 8-12 membered bicyclic rings. Each ring of the bicyclic carbocycle can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. Carbocycles include bicyclic molecules in which one, two, or three or more atoms are shared between the two rings. The term "fused carbocycle" refers to a bicyclic carbocycle in which each ring shares two adjacent atoms with another ring. Each ring of the fused carbocycle can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. In an exemplary embodiment, an aromatic ring (e.g., phenyl) can be fused with a saturated or unsaturated ring (e.g., cyclohexane, cyclopentane, or cyclohexene). When valence permits, any combination of saturated bicyclic, unsaturated bicyclic, and aromatic bicyclic rings is included in the definition of carbocycle. Exemplary "carbocycles" include cyclopentane, cyclohexane, bicyclo [2.2.1] heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo [4.2.0] oct-3-ene, naphthalene, and adamantane. Exemplary fused carbocycles include decahydronaphthalene, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene, and bicyclo[4.1.0]hept-3-ene. "Carbocycle" may be substituted at any position or positions capable of carrying a hydrogen atom.

[0467] As used herein, the term "carbocyclylalkyl" refers to an alkyl group substituted with a carbocyclyl group.

[0468] The term "carbonate" is art-recognized and refers to the group -OCO2-.

[0469] As used herein, the term "carboxyl" refers to a group represented by the formula -CO2H.

[0470] The term "cycloalkyl" includes substituted or unsubstituted non-aromatic monocyclic structures, preferably 4 to 8-membered rings, more preferably 4 to 6-membered rings. The term "cycloalkyl" also includes polycyclic ring systems having two or more rings, wherein two or more carbon atoms are shared by two adjacent rings, wherein at least one ring is a cycloalkyl group and the substituents (e.g., R 100) is connected to the cycloalkyl ring, for example, the other ring can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclic. Heteroaryl includes, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, benzodioxane, tetrahydroquinoline, etc.

[0471] As used herein, the term "ester" refers to the group -C(O)OR 9 , where R 9 It represents a hydrocarbon group.

[0472] As used herein, the term "ether" refers to a hydrocarbyl group connected to another hydrocarbyl group through an oxygen. Thus, an ether substituent of a hydrocarbyl group may be hydrocarbyl-O-. An ether may be symmetrical or asymmetrical. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethers include "alkoxyalkyl" groups, which may be represented by the general formula alkyl-O-alkyl.

[0473] As used herein, the terms "halo" and "halogen" refer to halogens, including chlorine, fluorine, bromine, and iodine.

[0474] As used herein, the terms "hetaralkyl" and "heteroaralk yl" refer to an alkyl group substituted with a heteroaryl group.

[0475] The terms "heteroaryl" and "heteroaryl" include substituted or unsubstituted aromatic monocyclic structures, preferably 5 to 7 rings, more preferably 5 to 6 rings, whose ring structure includes at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heteroaryl" and "heteroaryl" also include polycyclic systems with two or more rings, wherein two or more carbons are shared by two adjacent rings, wherein at least one ring is heteroaromatic, for example, other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclic radicals. Heteroaryl includes, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine etc.

[0476] As used herein, the term "heteroatom" means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen and sulfur.

[0477] As used herein, the term "heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclyl group.

[0478] The terms "heterocyclyl", "heterocycle" and "heterocyclic" refer to substituted or unsubstituted non-aromatic ring structures, preferably 3 to 10-membered rings, more preferably 3 to 7-membered rings, whose ring structures contain at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more rings, wherein two or more carbon atoms are common to two adjacent rings, wherein at least one ring is heterocyclic, for example, the other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclyl. Heterocyclyl includes, for example, piperidine, piperazine, pyrrolidine, morpholine, lactone, lactam, etc.

[0479] As used herein, the term "hydrocarbyl" refers to a group bonded by a carbon atom that does not have a =O or =S substituent, and typically has at least one carbon-hydrogen bond and a main chain that is mainly carbon, but may optionally contain heteroatoms. Therefore, for the purposes of this application, groups such as methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered to be hydrocarbyl, but substituents such as acetyl (which has a =O substituent on the connecting carbon) and ethoxy (which is connected by oxygen rather than carbon) are not. Hydrocarbyl includes, but is not limited to, aryl, heteroaryl, carbocycle, heterocycle, alkyl, alkenyl, alkynyl, and combinations thereof.

[0480] As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group.

[0481] When used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl or alkoxy, the term "low" means including groups having ten or fewer atoms, preferably six or fewer atoms in the substituent. For example, "low alkyl" refers to an alkyl group containing ten or fewer, preferably six or fewer carbon atoms. In certain embodiments, the acyl, acyloxy, alkyl, alkenyl, alkynyl or alkoxy substituents defined herein are respectively low acyl, low acyloxy, low alkyl, low alkenyl, low alkynyl or low alkoxy, whether they occur alone or in combination with other substituents, such as in narrating hydroxyalkyl and aralkyl (in this case, for example, when calculating the carbon atoms in the alkyl substituent, the atoms in the aryl are not calculated).

[0482] The terms "polycyclic group", "polycyclic" and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclyl) wherein two or more atoms are common to two adjacent rings, e.g., the ring is a "fused ring". Each ring of the polycyclic ring may be substituted or unsubstituted. In certain embodiments, each ring of the polycyclic ring contains 3 to 10 atoms, preferably 5 to 7 atoms, in the ring.

[0483] The term "sulfate" is art-recognized and refers to the group -OSO3H or a pharmaceutically acceptable salt thereof.

[0484] The term "sulfonamide" is art-recognized and refers to a group represented by the general formula

[0485]

[0486] Where R 9 and R 10 independently represent hydrogen or a hydrocarbon group.

[0487] The term "sulfoxide" is art-recognized and refers to the group -S(O)-.

[0488] The term "sulfonate" is art-recognized and refers to the group SO3H or a pharmaceutically acceptable salt thereof.

[0489] The term "sulfone" is art-recognized and refers to the group -S(O)2-.

[0490] The term "substituted" refers to a substituent having a substituent replacing hydrogen on one or more carbons of the main chain. It should be understood that "substituted" or "substituted by ... " includes implicit conditions, i.e., such substitution is based on the allowed valence of the substituted atom and the substituent, and the substitution produces a stable compound, for example, it will not spontaneously undergo transformations such as by rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" considers all allowed substituents of organic compounds. In a broad sense, allowable substituents include non-cyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Allowable substituents can be one or more substituents and are the same or different for appropriate organic compounds. For purposes of the present invention, heteroatoms such as nitrogen can have any allowable substituents that satisfy the valence of heteroatoms of hydrogen substituents and / or organic compounds described herein. Substituents may include any of the substituents described herein, for example, halogen, hydroxy, carbonyl (such as carboxyl, alkoxycarbonyl, formyl or acyl), thiocarbonyl (such as thioester, thioacetate or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfonamido, sulfonyl, heterocyclyl, aralkyl or aromatic or heteroaromatic moieties. Those skilled in the art will appreciate that the moieties substituted on the hydrocarbon chain may themselves be substituted if appropriate.

[0491] As used herein, the term "thioalkyl" refers to an alkyl group substituted with a thiol group.

[0492] As used herein, the term "thioester" refers to the group -C(O)SR 9 or –SC(O)R 9

[0493] Where R 9 It represents a hydrocarbon group.

[0494] As used herein, the term "thioether" is equivalent to an ether in which the oxygen is replaced by sulfur.

[0495] The term "urea" is art-recognized and can be represented by the general formula

[0496]

[0497] Where R 9 and R 10 independently represent hydrogen or a hydrocarbon group.

[0498] As used herein, the term "modulate" includes inhibiting or suppressing a function or activity (such as cell proliferation) as well as enhancing a function or activity.

[0499] The phrase "pharmaceutically acceptable" is recognized in the art. In certain embodiments, the term includes compositions, excipients, adjuvants, polymers and other materials and / or dosage forms that are suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic response or other problems or complications, and commensurate with a reasonable benefit / risk ratio, within the scope of sound medical judgment.

[0500] "Pharmaceutically acceptable salt" or "salt" is used herein to refer to acid addition salts or base addition salts that are suitable for use in or compatible with the treatment of a patient.

[0501] As used herein, the term "pharmaceutically acceptable acid addition salt" means any non-toxic organic or inorganic salt of any base compound represented by Formula I. Exemplary inorganic acids forming suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid and metal salts, such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Illustrative organic acids forming suitable salts include monocarboxylic acids, dicarboxylic acids and tricarboxylic acids, such as glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, benzoic acid, phenylacetic acid, cinnamic acid and salicylic acid and sulfonic acids, such as p-toluenesulfonic acid and methanesulfonic acid. Monobasic or dibasic acid salts can be formed, and such salts can exist in hydrated form, solvated form or substantially anhydrous form. Typically, the acid addition salts of the compound of Formula I are more soluble in water and various hydrophilic organic solvents, and typically exhibit a higher melting point compared to their free base forms. The selection of suitable salts is known to those skilled in the art. Other non-pharmaceutically acceptable salts, such as oxalates, may be used, for example, to isolate a compound of formula I for laboratory use, or for subsequent conversion to a pharmaceutically acceptable acid addition salt.

[0502] As used herein, the term "pharmaceutically acceptable base addition salt" means any nontoxic organic or inorganic base addition salt of any acid compound represented by Formula I or any intermediate thereof. Illustrative inorganic bases that form suitable salts include lithium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide or barium hydroxide. Exemplary organic bases that form suitable salts include aliphatic, alicyclic or aromatic organic amines, such as methylamine, trimethylamine and picoline or ammonia. The selection of appropriate salts will be known to those skilled in the art.

[0503] Many compounds that can be used for the methods and compositions of the present disclosure have at least one stereocenter in their structure. This stereocenter can exist in R or S configuration, and the R and S symbols are used according to the rules described in Pure Appli. Chem. (1976), 45, 11-30. The present disclosure contemplates all stereoisomeric forms, such as enantiomers and diastereoisomers (including all possible mixtures of stereoisomers) of compounds, salts, prodrugs, or mixtures thereof. See, for example, WO01 / 062726.

[0504] Additionally, certain alkenyl-containing compounds may exist as Z (same) or E (hetero) isomers. In each case, the disclosure includes both the mixture and the separate individual isomers.

[0505] Some compounds may also exist as tautomeric forms. Although not explicitly indicated in the formulas described herein, such forms are intended to be included within the scope of the present disclosure.

[0506] "Prodrug" or "pharmaceutically acceptable prodrug" refers to a compound that is metabolized (e.g., hydrolyzed or oxidized) in a host to form a compound of the present disclosure (e.g., a compound of formula I) after administration. Typical examples of prodrugs include compounds having biologically unstable or cleavable (protective) groups on the functional portion of the active compound. Prodrugs include compounds that can be oxidized, reduced, aminized, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated, acylated, deacylated, phosphorylated, or dephosphorylated to produce an active compound. Examples of prodrugs using esters or phosphoramidates as biologically unstable or cleavable (protective) groups are disclosed in U.S. Patents 6,875,751, 7,585,851, and 7,964,580, the disclosures of which are incorporated herein by reference. Prodrugs of the present disclosure are metabolized to produce compounds of formula I. The present disclosure includes within its scope prodrugs of the compounds described herein. Conventional procedures for the selection and preparation of suitable prodrugs are described, for example, in "Design of Prodrugs", Ed. H. Bundgaard, Elsevier, 1985.

[0507] As used herein, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition or vehicle that can be used to formulate a medicament for medical or therapeutic use, such as a liquid or solid filter aid, diluent, excipient, solvent or encapsulating material.

[0508] As used herein, the term "logarithm of solubility", "LogS" or "logS" is used in the art to quantify the water solubility of a compound. The water solubility of a compound significantly affects its absorption and distribution characteristics. Low solubility is often associated with poor absorption. The LogS value is the unit exfoliation logarithm (base 10) of the solubility measured in moles / liter.

[0509] As used herein, the phrase "expression level" refers to the expression level and / or prevalence of an expression product in a sample. For example, the expression level of a protein can be measured by staining a tissue sample (e.g., a plurality of cells) and measuring the prevalence (i.e., incidence) and / or level of the protein in one or more cells (preferably a plurality of cells) of the tissue or in the entire tissue sample.

[0510] Example

[0511] Having now generally described the invention, the invention will be more readily understood by reference to the following examples, which are included merely for the purpose of illustrating certain aspects and embodiments of the invention and are not intended to limit the invention.

[0512] Exemplary methods and compounds related to the present disclosure can be found, for example, in U.S. Pat. Nos. 10 / 160,753, 09 / 732,095, 10 / 758,518, published U.S. patent application No.: 20210290628, pending U.S. application No.: 17 / 680,995, and PCT application Nos. WO22 / 031330 and US21 / 59668; the contents of which are incorporated herein by reference in their entirety.

[0513] Embodiment 1: Exemplary Antibodies of the Disclosure

[0514] Table 1 describes exemplary antibodies of the present disclosure, relevant source information, recommended dilution levels for use in the protocols of Examples 2 and 3, and comments regarding expression patterns observed in certain embodiments of the present disclosure.

[0515]

[0516]

[0517] Embodiment 2: Exemplary Methods for Determining NF-κBp-p50 or NF-κBp-p65 Expression

[0518] Formalin-fixed, paraffin-embedded human tonsil and lymphoma sections were used. Tissue sections (5 μm) were deparaffinized and antigen retrieval was performed in citrate buffer at 90-100°C for 10-40 minutes. The sections were incubated in 1% catalase for 10 minutes to quench endogenous tissue peroxidase. The tissue sections were then incubated with a primary NF-κB p-p50 specific antibody (dilution 1:300) at 4°C for approximately 16 hours. The primary NF-κB p-p50 specific antibody used NF-κB p-p50 (S337), sc-271908 from Santa Cruz Biotechnology.

[0519] Slides were stained using a standard EnVision+ System-HRP kit (DAKO, Carpinteria, CA) according to the manufacturer's protocol. Immunohistochemistry was performed using diaminobenzidine as a chromogenic peroxidase substrate, and slides were counterstained with hematoxylin. Negative control samples included replacement of the primary antibody with non-immune IgG1 (Dako).

[0520] At a 1:100 dilution of NF-κB p-p50 Ab, specific staining of the target molecule with low background staining was observed in human tonsil and lymphoma samples ( Fig. 12C ). Specifically, nuclear and / or cytoplasmic expression of NF-κB p-p50 was found in all 6 SD cases treated with 50 mg QD (2 cases, tumor regression), 50 mg BID (1 case), 200 BID (1 case, tumor regression), and 400 BID (2 cases). Expression of NF-κB p-p50 was not detected in 7 of 8 PD patients, including patients treated with 50 mg QD (1 case), 100 mg QD (1 case), 100 mg BID (3 cases), 200 mg BID (1 case), and 400 BID (1 case). A statistically significant correlation was observed between the expression of NF-κB p-p50 in tumor biopsies and SD in NHL patients treated with compound 1 (p<0.05). Analysis of NF-κB p-p50 expression in paired tumor biopsies (n=3) collected before and after Compound 1 treatment showed that NF-κB p-p50 expression was significantly downregulated in tumors obtained from NHL patients treated with Compound 1. In vitro experiments demonstrated expression of NF-κB p-p50 in 3D lymphoma organoids treated with clinically relevant concentrations of Compound 1. Our results support further development of NF-κB p-p50 as a potential predictive and pharmacodynamic biomarker for IRAK4 inhibitors.

[0521] In conclusion, the expression of NF-κB p-p50 can be used as a biomarker to predict SD in NHL patients in response to treatment with IRAK4-modifying compounds. The NF-κB p-p50 selection strategy can be used in future clinical trials to identify NHL patients who are most likely to respond to treatment with IRAK4-modifying compounds in combination with chemotherapy or targeted therapeutics.

[0522] Embodiment 3: For evaluation of antibody expression in FFPE (formalin-fixed paraffin-embedded) AML bone marrow agglutinate samples Exemplary Single IHC Staining Protocol

[0523] The tissue sections were dewaxed three times in xylene for 5 minutes each time. The dewaxed tissue sections were washed twice in 100% ethanol for 5 minutes each time and further washed in 95% ethanol for 5 minutes. The resulting sections were washed in tap water for 5 minutes. The washed tissue sections were incubated in a dual endogenous peroxidase blocker (Dako Envision, K4065) for 10 minutes and then washed again in tap water for 5 minutes.

[0524] The slides were placed in a container and covered with citrate buffer (Cat. No. C9999-1000ML, MilliporeSigma), heated in a microwave oven while maintaining a sufficient amount of citrate buffer to completely cover the slides. The slides were then cooled in the buffer at ambient temperature. The slides were washed three times in 1X wash buffer (Envision Flex wash buffer, A gilent, K800721-2) for 5 minutes each time.

[0525] The slides were incubated overnight at 4°C in a humidified chamber with a primary antibody (selected from the antibodies listed in Table 1). The incubated slides containing the primary antibody were washed three times in a wash buffer for 5 minutes each time. The labeled polymer-HRP solution (DAKO EnVision+System-HRP) was applied to the slides and the slides were incubated at room temperature for 30 minutes. The resulting slides were washed three times in phosphate buffered saline for 5 minutes each time. The substrate-chromogen solution was prepared by mixing 1 mL of substrate buffer with 20 μL of liquid DAB+ chromogen (DAB+=DAKO EnVision Dual Link System-HRP, catalog number K4065). The resulting substrate-chromogen solution was applied to the prepared tissue sections for staining for 5 minutes. The slides were then washed in tap water for 5 minutes to stop staining. The washed slides were counterstained with freshly filtered Mayer's hematoxylin for 5 seconds and washed in tap water for 10 minutes. Slides were washed repeatedly three times with 100% ethanol for 5 min each and placed in xylene for 5 s before mounting and covering with a glass coverslip.

[0526] Embodiment 4: Performance of compound 1 in WM

[0527] The patient is an otherwise healthy male, 49 years of age, who complained of severe fatigue. Routine labs were notable for an elevated erythrocyte sedimentation rate and anemia; therefore, he was referred to the hematology / oncology department. Further workup revealed IgMλm-protein on serum protein electrophoresis and bone marrow hypercellularity with trilineage hematopoiesis and an atypical lymphoplasmacytic infiltration, consistent with WM. A CT scan did not reveal lymphadenopathy or hepatosplenomegaly.

[0528] Because of symptomatic cytopenias and profound fatigue, treatment was recommended. Between 2005 and 2007, the patient received 375 mg / m 2Rituximab was induced and continued for 8 weeks with IV, followed by maintenance rituximab every 3 months for 8 doses, achieving a very good partial remission. He did well for approximately 4 years, at which time his disease progressed and he developed recurrent symptomatic anemia and a new grade 1 sensory peripheral neuropathy involving the hands and feet. Between June and September 2011, he was retreated with rituximab with stable disease, during which time the IgM value increased from 1476 mg / dL to 2042 mg / dL, and no improvement in symptoms. In November 2011, a repeat bone marrow biopsy showed a 90% cellular bone marrow with WM accounting for 20% of the cellularity, IgM lambda plasma cells accounting for 5%-10% of the cellularity, normal cytogenetics, no increase in reticulin staining, and trace amounts of stainable iron. At this time, there was no lymphadenopathy or organomegaly on CT scan. By December 2012, his serum IgM increased to 3380 mg / dL and IgMλm-protein was 2.37 g / dL. He was referred to a tertiary care center for further treatment, where induction chemotherapy followed by autologous stem cell transplantation was recommended. In early 2013, he received 2 cycles of rituximab, cyclophosphamide, bortezomib, and dexamethasone (R-CyBorD), achieving a partial remission (PR) with a decrease in IgM to 1285 mg / dL and m-protein to 0.88 g / dL. Then in June 2013, the patient received rituximab, ifosfamide, etoposide chemomobilization, and stem cell collection, after which his IgM and m-protein remained unchanged. Two cycles of bendamustine and rituximab (BR) were administered before transplantation to further reduce the cells, which deepened the patient's partial response (IgM 454 mg / dL, m-protein 0.30 g.dL). In October 2013, he underwent autologous stem cell transplantation with BEAM conditioning (carmustine, etoposide, cytarabine, melphalan) without complications and achieved a very good partial response (VGPR) with a post-transplant IgM nadir of 135 mg / dL and m-protein detectable by immunofixation only in January 2014.

[0529] The patient remained asymptomatic for more than 4 years, during which time m-protein and IgM slowly increased. By mid-2017, he began to experience increasing fatigue. In November 2017, bone marrow evaluation revealed a normocellular bone marrow with 30% involvement of WM and normal cytogenetics. Next-generation sequencing revealed the RICTOR N1065S mutation in a subclonal population as well as the MYD88 L265P mutation and TET2 mutation. There was no evidence of CXCR4 genomic alterations. By the end of 2018, his fatigue had begun to interfere with his ability to perform daily activities, so treatment was again recommended. Several options were discussed, including clinical trials and standard of care Bruton's tyrosine kinase inhibitor (BTKi) therapy. Given his clinical history, current symptoms, known mutational profile, and personal preferences, he was enrolled in a phase 1, dose-escalation study of a novel oral IRAK4 inhibitor (Compound 1) in patients with relapsed or refractory B-cell malignancies (NCT03328078).

[0530] Baseline testing in December 2018 included a bone marrow biopsy that showed 5%-10% WM involvement, m-protein of 1.66 g / dL, IgM of 2,801 mg / dL, and no pathological lymphadenopathy or hepatosplenomegaly on computed tomography. Quantitative immunoglobulins and serum protein electrophoresis were obtained each cycle to determine response to treatment ( Figure 1 ).

[0531] The patient started treatment at the first dose level of 50 mg. He tolerated the treatment well with no adverse events. During the first six 21-day cycles, his m-protein slowly but steadily tended to decline to 1.55 g / dL, and IgM initially increased from 2801 mg / dL to 2866 mg / dL during the first 2 cycles, then decreased to 2639 mg / dL on day 1 of cycle 6 ( Figure 1). Using a standard 3+3 design, subsequent dose levels, 100 mg orally twice daily, were cleared per protocol. Given the lack of evidence of response with significant toxicity, the patient was a candidate for escalation to 100 mg orally twice daily starting on Day 1 of Cycle 7 in April 2019. During Cycles 7 and 8, he continued to have stable disease (SD) with a downward trend in IgM and m-protein with no evidence of toxicity and thus became a candidate for escalation to the next cleared dose level of 200 mg orally twice daily starting on Day 1 of Cycle 9 in May 2019. Prior to the dose escalation to 200 mg twice daily, his baseline IgM was 2245 mg / dL and m-protein was 1.37 g / dL. His baseline symptoms of fatigue improved slightly but persisted. At the dose level of 200 mg orally twice daily, he continued to enjoy a seemingly dose-dependent reduction in tumor markers and his fatigue had completely resolved by the summer of 2019. He reported a significant improvement in his quality of life, elimination of fatigue, and resumed his rigorous daily exercise program that he had been unable to perform for the past 2 years due to WM-related symptoms. In August 2019, he was found to have an asymptomatic grade 2 creatine phosphokinase (CPK) elevation, and extensive examinations and physical examinations did not reveal any problems. He reduced the intensity of his exercise program, and the asymptomatic CPK elevation was completely eliminated without the need for compound 1 dose delay or reduction. During the 15th to 20th cycles, the patient's IgM stabilized at around 1500mg / dL and m-protein stabilized at around 0.9g / dL when at a dose of 200mg orally twice a day. Based on the safety data evolving at higher dose levels, the patient was increased to 300mg orally twice a day starting from the 20th cycle in January 2020. He once again experienced a seemingly dose-dependent acceleration of the response without significant toxicity, and achieved PR on the first day of the 22nd cycle (m-protein 0.68g / dL, IgM 1241mg / dL). He continued to take Compound 1, 300 mg orally twice daily.

[0532] The treatment of WM involves targeting pathways associated with known mutations of MYD88 and CXCR-4. Previous studies have shown the role of IRAK4 in signal cascades, which participate in the stimulation of proinflammatory cytokines by forming a complex with MYD88. Therefore, IRAK4 is an essential component for regulating immune responses, and those with dysfunction in any part of the complex can lead to immunodeficiency or immune dysregulation. With the addition of IRAK4 inhibitors, a strong association is formed between IRAK4 and MYD88, and a weak association is formed with IRAK-1, thereby reducing the ubiquitination of IRAK1, ultimately leading to a reduction in IL-1-induced signaling and cytokine production.

[0533] By inhibiting IRAK4, compound 1 prevents NF-κB activation, resulting in reduced production of inflammatory cytokines and potential anti-tumor, immunomodulatory and anti-inflammatory effects. Preclinical studies have also shown that compound 1 affects TLR / IL1R signaling, which may prevent inflammatory processes in autoimmune disorders.

[0534] The patient tolerated continuous oral treatment with Compound 1 well for nearly 18 months. His tumor burden has been shrinking in a dose-dependent manner, achieving a partial response status (PR) ( Figure 1 ). The patient's quality of life improved from baseline with resolution of fatigue, even allowing him to engage in the vigorous physical exercise he had enjoyed prior to disease onset. However, this resulted in intermittent asymptomatic Grade 2 elevations in CK that were resolved by moderate exercise without the need to maintain or reduce Compound 1 exposure.

[0535] Embodiment 5: Performance of Compound 1 in DLBCL, FL, HGBL, WM, LPL, MZL, and MCL

[0536] Research Design and Methods

[0537] Phase I trial compound 1 is a dose escalation trial using a 3+3 design. Seven dosing groups include 50mg and 100mg QD and 50mg, 100mg, 200mg, 300mg or 400mg BID of daily continuous oral monotherapy in a 21-day cycle. Targets include safety and tolerability (primary), pk / pd and early efficacy (secondary) and biomarker correlation (exploratory). 31 patients with drug-resistant or refractory advanced NHL have been recruited. The detailed information of the patient population is shown in Table 2 below.

[0538] Table 2 - Summary of adverse effects

[0539]

[0540]

[0541] *High-grade composite low-high-grade disease based on local pathology report

[0542] **Includes lymphoplasmacytic cells (n=2), mantle cells (n=2), marginal zone (n=2), high-grade MYC-BCL6 (n-1)

[0543] result

[0544] Compound 1 was well tolerated. Eight patients were exposed at the highest dose level of 400 mg twice a day: 2 of 5 DLT evaluable patients had grade 3 rhabdomyolysis (DLT), without complications, and reversible after treatment interruption and hydration / analgesia therapy-then all continued treatment at a lower dose of 200 mg twice a day or 300 mg twice a day, respectively. Six patients tolerated 300 mg twice a day well without DLT. Most non-hematological TEAEs were grade 1 or 2 and manageable, including diarrhea, vomiting, fatigue, dyspnea, and myalgia. Mild / moderate, neutropenia, anemia, thrombocytopenia; at dose levels within the range of 200 mg twice a day and 400 mg twice a day, only 4 of 18 patients had grade 3 combined attacks but no complications (Table 3). There was no toxic death. Pharmacokinetics showed favorable characteristics, with a proportional increase in dose when exposed. Similar pharmacodynamic changes were shown in terms of cytokine reduction. Duration of treatment ranged from <1 month to 18+ months with sustained disease control. Eight of 28 evaluable patients experienced a ≥20% reduction in overall tumor burden from baseline, with greater reductions at higher doses (Table 5). WM patients with sustained PR underwent intrapatient dose escalation with a dose / response relationship and very good treatment tolerability ( Fig. 20 ). Downstream pharmacodynamic markers of IRAK4 and molecular characteristics including cell sources will be shown.

[0545] Table 3 - Summary of adverse effects

[0546]

[0547] Table 4 - Summary of Adverse Hematological Effects

[0548]

[0549]

[0550] Table 5 - Reaction Overview

[0551]

[0552] In summary, compound 1 exhibited good safety and tolerability, desirable pharmacokinetic properties, and preliminary clinical activity.

[0553] Embodiment 6: Performance of compound 1 in AML and MDS

[0554] Research Design and Methods

[0555] This is a single-arm dose-escalation Phase 1 study of orally administered Compound 1 monotherapy in adult patients with AML or high-risk MDS (NCT04278768). The study was conducted in 2 parts: an initial dose escalation and a dose expansion phase. The starting dose level is 200 mg twice a day, which has been determined to be safe, capable of achieving relevant levels of drug exposure and showing signs of biological activity and clinical efficacy in NHL studies. Three patients with AML or MDS will be recruited at the designated dose. If the first 3 patients do not experience DLT during the first cycle, patients can be recruited to the next higher dose level of 300 mg twice a day until a safe and effective RP2D is established.

[0556] The study is expected to recruit approximately 18 patients to establish an initial RP2D. The safety population will include all patients who receive any dose of compound 1 in the study, and the efficacy population will include patients with valid baseline and post-baseline disease assessments and who receive at least one dose of study drug. Each treatment cycle of compound 1 will be 28 days in length and repeated in the absence of toxicity or disease progression.

[0557] The main study inclusion and exclusion criteria were as follows: relapsed or refractory AML (primary or secondary, including treatment-related) after at least 6 cycles of hypomethylating agents [HMA] or evidence of early progression based on investigator assessment or high / very high risk relapsed / refractory MDS (IPSS-R criteria). Patients diagnosed with acute promyelocytic leukemia (APL, M3), CML blast, allogeneic hematopoietic stem cell transplant (Allo-HSCT) within 60 days of the first dose of Compound 1, or clinically significant graft-versus-host disease (GVHD) requiring continued up-titration of immunosuppressive drugs before starting Compound 1 were excluded.

[0558] The primary objective is to determine the maximum tolerated dose (MTD) and recommended phase 2 dose (RP2D) of compound 1 in patients with AML and high-risk MDS based on safety and tolerability, DLT, and PK / PD results.

[0559] result

[0560] All naive patients completed cycle 1 with a reduction in bone marrow blasts, including several complete bone marrow responses.

[0561] Group 1 (200 mg twice daily; cycle duration 4 weeks)

[0562] 3 patients with hr-MDS; all on ongoing treatment (currently 2 cycles - 4 cycles). Cycle 1 no DLT.

[0563] One patient had a C2 reduction in Gr dose. Three patients had dizziness.

[0564] Group 2 (300 mg twice a day)

[0565] 4 patients (3 AML, 1 hr-MDS), all are on treatment (currently 1-2 cycles. No DLT in the first 3 patients).

[0566] Group 3 open (400 mg twice a day)

[0567] Open for solicitation.

[0568] Embodiment 7: Performance of Compound 1 in Relapsed or Refractory Hematological Malignancies

[0569] Research Design and Methods

[0570] This is a trial of the combination of compound 1 and ibrutinib orally administered in adult patients with relapsed or refractory hematological malignancies. (NCT03328078). It has 2 parts: the initial dose escalation phase (Part A2) and the expansion part of 4 groups. In the 3x3 dose escalation design, the starting oral dose of compound 1 will be 200 mg BID, administered daily. At the same time, patients receive a labeled dose of ibrutinib (560 mg or 420 mg) for the corresponding NHL subtype daily. If well tolerated, the dose of compound 1 will be escalated to 300 mg BID. Goals include safety / tolerance, pharmacokinetics, preliminary efficacy assessments, and exploratory biomarker correlations. Once the recommended Phase 2 dose (RP2D) for the combined dose is determined, the expansion phase (Part B) will evaluate the efficacy (CR / ORR rate / duration), safety / tolerance, population PK, and biomarker correlation of the combination of compound 1 and ibrutinib. Part B will contain four cohorts including: 1-MZL, 2-DLBCL, 3-CNSL, and 4-NHL with adaptive ibrutinib resistance (basket design).

[0571] Groups 1-3 must not use BTK inhibitors. The latter group should have received and responded to ibrutinib monotherapy (no primary resistance). Once they develop adaptability, secondary resistance and show tumor progression, a combination of ibrutinib and compound 1 will be given. (A short interval of <3 weeks of ibrutinib therapy is acceptable.) This group will include patients with ibrutinib-approved or NCCN-recommended indications: MCL, MZL, CLL / SLL, WM / LPL, PCNSL (NCCN listed).

[0572] Primary Objectives: Identification of preliminary efficacy signals of improved objective responses in Cohorts 1-3 compared to historical data and demonstration of reversal of resistance in Cohort 4 by showing an objective response after prior progression.

[0573] The estimated sample size of up to about 18 patients in Part A2 is based on the standard 3+3 study design for dose escalation. When co-administered with ibrutinib, the exact number of patients will be determined by establishing the maximum tolerated dose (MTD) of compound 1 and the number of groups required for the recommended Phase 2 dose (RP2D). For Part B dose expansion, up to 46 patients are recruited in each group of 4 NHL groups. The safety population will include all patients who receive any dose of the combination of compound 1 and ibrutinib in the study, and the efficacy population will include patients with effective baseline and post-baseline disease assessment and receiving at least one dose of the study combination drug. Safety observations and measurements include drug exposure, AE, safety laboratory tests, vital signs, physical examinations, ECG and ECOG performance status. Each treatment cycle length of compound 1 will be 21 days, and repeated in the absence of toxicity or tumor progression, and ibrutinib will be administered according to the label.

[0574] The primary study inclusion and exclusion criteria for the combination therapy dose escalation part A2 are as follows: A diagnosis of histopathologically confirmed B-cell NHL according to the WHO 2016 classification. Eligible NHL subtypes include follicular lymphoma, MZL, mantle cell lymphoma, DLBCL (including extranodal lymphoma of the leg, testis, or NOS type), CLL / SLL, primary or secondary CNS lymphoma, and Waldenstrom's macroglobulinemia / LPL. Patients with mantle cell lymphoma, MZL, WM / LPL, or CLL / SLL should meet clinical criteria for treatment of their disease. Patients with the presence of acute or chronic toxicity (except alopecia) caused by previous anticancer therapy that has not resolved to ≤ Grade 1 as determined by NCI CTCAE v4.03 within 7 days before the start of the study will be excluded.

[0575] The endpoints of the study treatment are to determine the safety and tolerability, DLT, MTD and RP2D of the combination of oral compound 1 and ibrutinib, and the secondary endpoints are to evaluate the objective response rate (ORR), durable response rate (DOR) DCR, PFS and OS after treatment with the combination of compound 1 and ibrutinib.

[0576] Embodiment 8: Performance of Compound 1 in Autoimmune Diseases

[0577] Compound 1 will be administered to subjects with autoimmune disorders (eg, graft-versus-host disease) in a dose escalation study starting at 50 mg. The efficacy of Compound 1 will be determined by methods known to those of ordinary skill in the art.

[0578] Embodiment 9: Exemplary treatment of OCL-LY10 and TF-1 cells with compound 1

[0579] OCL-LY10 and TF-1 cells were treated with different concentrations of 3 μM and 10 μM compound 1. 48 hours after treatment, cell lysates were obtained. Protein sample concentrations were quantified, and equal amounts of 20 μg of whole protein extracts were loaded into each well of SDS-polyacrylamide gels. Cell extracts were separated by 10% SDS-PAGE, transferred to nitrocellulose membranes, and probed as indicated. The following antibodies were used for immunoblot analysis: NF-κB p-p50S337 (Santa Cruz Biotechnology) and b-actin (Cell Signaling Technology). The expression of NF-κB p-p50 S337 was downregulated in OCL-Ly10 and TF-1 cell lines treated with compound 1.

[0580] Incorporated by Reference

[0581] All publications and patents mentioned herein are hereby incorporated by reference in their entirety to the same extent as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In the event of a conflict, the present application, including any definitions herein, will control.

[0582] Equivalent solutions

[0583] Although specific embodiments of the present invention have been discussed, the above description is illustrative and non-restrictive. After reading this specification and the appended claims, many variations of the present invention will become apparent to those skilled in the art. The full scope of the present invention should be determined by reference to the full scope of the claims together with their equivalents, and this specification together with such variations.

Claims

1. A method of treating a disease or condition in a subject, the method comprising: obtaining one or more biological samples from the subject; analyzing the biological sample for the presence of a first biomarker in nuclei of cells of the biological sample; and administering to the subject an IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader when the presence of the first biomarker is detected in the nucleus of the cell of the biological sample; The first biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

2. The method of claim 1, wherein the method further comprises: analyzing the biological sample for the presence of a second biomarker in nuclei of cells of the biological sample; administering the IRAK4 modifying compound selected from the IRAK4 inhibitor or the IRAK4 degrader to the subject only when the presence of the first biomarker and the second biomarker is detected in the nucleus of the cell of the biological sample; and Wherein the second biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

3. The method of claim 2, wherein the method further comprises: analyzing the biological sample for the presence of a third biomarker in nuclei of cells of the biological sample; and administering the IRAK4 modifying compound selected from the IRAK4 inhibitor or the IRAK4 degrader to the subject only when the presence of the first biomarker, the second biomarker, and the third biomarker is detected in the nucleus of the cell of the biological sample; wherein the third biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

4. The method of claim 3, wherein the method further comprises: analyzing the biological sample for the presence of a fourth biomarker in nuclei of cells of the biological sample; and administering the IRAK4 modifying compound selected from the IRAK4 inhibitor or the IRAK4 degrader to the subject only when the presence of the first biomarker, the second biomarker, the third biomarker, and the fourth biomarker is detected in the nucleus of the cell of the biological sample; wherein the fourth biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

5. A method of treating a disease or condition in a subject, the method comprising: obtaining one or more biological samples from the subject; measuring a sample expression level of a first biomarker in the biological sample; comparing the sample expression level of the first biomarker to a reference expression level of the first biomarker; administering to the subject an IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader when the sample expression level of the first biomarker in the sample is elevated compared to the reference expression level of the first biomarker; and The first biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

6. The method of claim 5, wherein the method further comprises: measuring the sample expression level of a second biomarker in the biological sample; comparing the sample expression level of the second biomarker to a reference expression level of the second biomarker; as well as administering the IRAK4 modifying compound selected from the IRAK4 inhibitor or the IRAK4 degrader to the subject only if the sample expression levels of the first marker and the second biomarker in the sample are each elevated as compared to the reference expression levels of the first marker and the second biomarker; Wherein the second biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

7. The method of claim 6, wherein the method further comprises: measuring the sample expression level of a third biomarker in the biological sample; comparing the sample expression level of the third biomarker to a reference expression level of the third biomarker; as well as administering the IRAK4 modifying compound selected from the IRAK4 inhibitor or the IRAK4 degrader to the subject only if the sample expression levels of the first biomarker, the second biomarker, and the third biomarker in the sample are each elevated compared to the reference expression levels of the first biomarker, the second biomarker, and the third biomarker; wherein the third biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

8. The method of claim 7, wherein the method further comprises: measuring the sample expression level of a fourth biomarker in the biological sample; comparing the sample expression level of the fourth biomarker to a reference expression level of the fourth biomarker; as well as administering an IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader to the subject only if the sample expression levels of the first biomarker, the second biomarker, the third biomarker, and the fourth biomarker in the sample are each elevated compared to the reference expression levels of the first biomarker, the second biomarker, the third biomarker, and the fourth biomarker; wherein the fourth biomarker is selected from IRAK4, NF-κBp-p65, NF-κBp-p50, p-IRAK1, FLT3 and MYD88.

9. A method of treating a disease or condition in a subject, the method comprising: obtaining one or more biological samples from the subject; measuring a sample expression level of a first biomarker in the biological sample; comparing the sample expression level of the first biomarker to a reference expression level of the first biomarker; analyzing the biological sample for the presence of the first biomarker in nuclei of cells of the biological sample; and An IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader is administered to the subject only if: detecting the presence of the first biomarker in the nucleus of the cell of the biological sample; as well as the sample expression level of the first biomarker in the sample is increased compared to the reference expression level of the first biomarker; The first biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

10. The method of claim 9, wherein the method further comprises: measuring the sample expression level of a second biomarker in the biological sample; comparing the sample expression level of the second biomarker to a reference expression level of the second biomarker; analyzing the biological sample for the presence of the second biomarker in nuclei of cells of the biological sample; and An IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader is administered to the subject only if: detecting the presence of each of the first biomarker and the second biomarker in the nucleus of the cell of the biological sample; as well as the sample expression levels of the first biomarker and the second biomarker in the sample are both increased compared to the reference expression levels of the first biomarker and the second biomarker; Wherein the second biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

11. The method of claim 10, wherein the method further comprises: measuring the sample expression level of a third biomarker in the biological sample; comparing the sample expression level of the third biomarker to a reference expression level of the third biomarker; analyzing the biological sample for the presence of the third biomarker in nuclei of cells of the biological sample; and An IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader is administered to the subject only if: detecting the presence of each of the first biomarker, the second biomarker, and the third biomarker in the nucleus of the cell of the biological sample; as well as the sample expression level of each of the first biomarker, the second biomarker, and the third biomarker in the sample is elevated compared to the reference expression level of each of the first biomarker, the second biomarker, and the third biomarker; wherein the third biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

12. The method of claim 11, wherein the method further comprises: measuring the sample expression level of a fourth biomarker in the biological sample; comparing the sample expression level of the fourth biomarker to a reference expression level of the fourth biomarker; analyzing the biological sample for the presence of the fourth biomarker in nuclei of cells of the biological sample; and An IRAK4 modifying compound selected from an IRAK4 inhibitor or an IRAK4 degrader is administered to the subject only if: detecting the presence of each of the first biomarker, the second biomarker, the third biomarker, and the fourth biomarker in the nucleus of the cell of the biological sample; as well as the sample expression level of each of the first biomarker, the second biomarker, the third biomarker, and the fourth biomarker in the sample is elevated compared to the reference expression level of each of the first biomarker, the second biomarker, the third biomarker, and the fourth biomarker; wherein the fourth biomarker is selected from IRAK4, NF-κB p-p65, NF-κB p-p50, p-IRAK1, FLT3 and MYD88.

13. The method of any one of claims 1 to 12, wherein the first biomarker is IRAK4.

14. The method of any one of claims 1 to 12, wherein the first biomarker is NF-κB p-p65.

15. The method of any one of claims 1 to 12, wherein the first biomarker is NF-κB p-p50.

16. The method of any one of claims 1 to 12, wherein the first biomarker is p-IRAK1.

17. The method of any one of claims 1 to 12, wherein the first biomarker is FLT3.

18. The method of any one of claims 1 to 12, wherein the first biomarker is MYD88.

19. The method of any one of claims 2 to 4, 6 to 8, 10 to 18, wherein the second biomarker is IRAK4.

20. The method of any one of claims 2 to 4, 6 to 8, 10 to 18, wherein the second biomarker is NF-κB p-p65.

21. The method of any one of claims 2 to 4, 6 to 8, 10 to 18, wherein the second biomarker is NF-κB p-p50.

22. The method of any one of claims 2 to 4, 6 to 8, 10 to 18, wherein the second biomarker is p-IRAK1.

23. The method of any one of claims 2 to 4, 6 to 8, 10 to 18, wherein the second biomarker is FLT3.

24. The method of any one of claims 2-4, 6-8, 10-18, wherein the second biomarker is MYD88.

25. The method of any one of claims 3, 4, 7, 8, 11-24, wherein the third biomarker is IRAK4.

26. The method of any one of claims 3, 4, 7, 8, 11-24, wherein the third biomarker is NF-κB p-p65.

27. The method of any one of claims 3, 4, 7, 8, 11-24, wherein the third biomarker is NF-κB p-p50.

28. The method of any one of claims 3, 4, 7, 8, 11-24, wherein the third biomarker is p-IRAK1.

29. The method of any one of claims 3, 4, 7, 8, 11-24, wherein the third biomarker is FLT3.

30. The method of any one of claims 3, 4, 7, 8, 11-24, wherein the third biomarker is MYD88.

31. The method of any one of claims 4, 8, 12 to 30, wherein the fourth biomarker is IRAK4.

32. The method of any one of claims 4, 8, 12 to 30, wherein the fourth biomarker is NF-κBp-p65.

33. The method of any one of claims 4, 8, 12 to 30, wherein the fourth biomarker is NF-κBp-p50.

34. The method of any one of claims 4, 8, 12-30, wherein the fourth biomarker is p-IRAK1.

35. The method of any one of claims 4, 8, 12 to 30, wherein the fourth biomarker is FLT3.

36. The method of any one of claims 4, 8, 12-30, wherein the fourth biomarker is MYD88.

37. The method of any one of claims 5 to 36, wherein the sample expression level of the first biomarker is indicative of the amount of nuclear expression of the first biomarker.

38. The method of any one of claims 5 to 37, wherein the reference expression level of the first biomarker is indicative of an amount of nuclear expression of the first biomarker.

39. The method of any one of claims 5 to 38, wherein the sample expression level of the first biomarker is indicative of the amount of cytoplasmic expression of the first biomarker.

40. The method of any one of claims 5 to 39, wherein the reference expression level of the first biomarker is indicative of a cytoplasmic expression amount of the first biomarker.

41. The method of any one of claims 5 to 40, wherein the sample expression level of the first biomarker is indicative of the amount of membrane expression of the first biomarker.

42. The method of any one of claims 5 to 41, wherein the reference expression level of the first biomarker is indicative of the amount of membrane expression of the first biomarker.

43. The method of any one of claims 6 to 8 and 10 to 42, wherein the sample expression level of the second biomarker is indicative of the amount of nuclear expression of the second biomarker.

44. The method of any one of claims 6 to 8 and 10 to 43, wherein the reference expression level of the second biomarker is indicative of an amount of nuclear expression of the second biomarker.

45. The method of any one of claims 6 to 8 and 10 to 44, wherein the sample expression level of the second biomarker is indicative of the amount of cytoplasmic expression of the second biomarker.

46. ​​The method of any one of claims 6 to 8 and 10 to 45, wherein the reference expression level of the second biomarker is indicative of the cytoplasmic expression amount of the second biomarker.

47. The method of any one of claims 6 to 8 and 10 to 46, wherein the sample expression level of the second biomarker is indicative of the amount of membrane expression of the second biomarker.

48. The method of any one of claims 6 to 8 and 10 to 47, wherein the reference expression level of the second biomarker is indicative of the amount of membrane expression of the second biomarker.

49. The method of any one of claims 7, 8, and 11 to 48, wherein the sample expression level of the third biomarker is indicative of an amount of nuclear expression of the third biomarker.

50. The method of any one of claims 7, 8, and 11 to 49, wherein the reference expression level of the third biomarker is indicative of an amount of nuclear expression of the third biomarker.

51. The method of any one of claims 7, 8, and 11 to 50, wherein the sample expression level of the third biomarker is indicative of a cytoplasmic expression amount of the third biomarker.

52. The method of any one of claims 7, 8, and 11 to 51, wherein the reference expression level of the third biomarker is indicative of a cytoplasmic expression amount of the third biomarker.

53. The method of any one of claims 7, 8, and 11 to 52, wherein the sample expression level of the third biomarker is indicative of the amount of membrane expression of the third biomarker.

54. The method of any one of claims 7, 8, and 11 to 53, wherein the reference expression level of the third biomarker is indicative of a membrane expression amount of the third biomarker.

55. The method of any one of claims 8 and 12 to 54, wherein the sample expression level of the fourth biomarker is indicative of the amount of nuclear expression of the fourth biomarker.

56. The method of any one of claims 8 and 12 to 55, wherein the reference expression level of the fourth biomarker is indicative of an amount of nuclear expression of the fourth biomarker.

57. The method of any one of claims 8 and 12 to 56, wherein the sample expression level of the fourth biomarker is indicative of a cytoplasmic expression amount of the fourth biomarker.

58. The method of any one of claims 8 and 12 to 57, wherein the reference expression level of the fourth biomarker is indicative of a cytoplasmic expression amount of the fourth biomarker.

59. The method of any one of claims 8 and 12 to 58, wherein the sample expression level of the fourth biomarker is indicative of the amount of membrane expression of the fourth biomarker.

60. The method of any one of claims 8 and 12 to 59, wherein the reference expression level of the fourth biomarker is indicative of a membrane expression amount of the fourth biomarker.

61. The method of any one of claims 5 to 60, wherein the sample expression level of IRAK4 is indicative of the amount of nuclear expression of IRAK4.

62. The method of any one of claims 5 to 61, wherein the reference expression level of IRAK4 indicates the amount of nuclear expression of IRAK4.

63. The method of any one of claims 5 to 62, wherein the sample expression level of IRAK4 indicates the amount of cytoplasmic expression of IRAK4.

64. The method of any one of claims 5 to 63, wherein the reference expression level of IRAK4 indicates the cytoplasmic expression amount of IRAK4.

65. The method of any one of claims 5 to 64, wherein the sample expression level of NF-κB p-p65 is indicative of the amount of nuclear expression of NF-κB p-p65.

66. The method of any one of claims 5 to 65, wherein the reference expression level of NF-κB p-p65 indicates the amount of nuclear expression of NF-κB p-p65.

67. The method of any one of claims 5 to 66, wherein the sample expression level of NF-κB p-p65 indicates the amount of cytoplasmic expression of NF-κB p-p65.

68. The method of any one of claims 5 to 67, wherein the reference expression level of NF-κB p-p65 indicates the cytoplasmic expression amount of NF-κB p-p65.

69. The method of any one of claims 5 to 68, wherein the sample expression level of NF-κB p-p50 is indicative of the amount of nuclear expression of NF-κB p-p50.

70. The method of any one of claims 5 to 69, wherein the reference expression level of NF-κB p-p50 indicates the amount of nuclear expression of NF-κB p-p50.

71. The method of any one of claims 5 to 70, wherein the sample expression level of NF-κB p-p50 is indicative of the amount of cytoplasmic expression of NF-κB p-p50.

72. The method of any one of claims 5 to 71, wherein the reference expression level of NF-κB p-p50 indicates the amount of cytoplasmic expression of NF-κB p-p50.

73. The method of any one of claims 5 to 72, wherein the sample expression level of p-IRAK1 is indicative of the amount of nuclear expression of p-IRAK1.

74. The method of any one of claims 5 to 73, wherein the reference expression level of p-IRAK1 indicates the amount of nuclear expression of p-IRAK1.

75. The method of any one of claims 5 to 74, wherein the sample expression level of p-IRAK1 is indicative of the amount of cytoplasmic expression of p-IRAK1.

76. The method of any one of claims 5 to 75, wherein the reference expression level of p-IRAK1 indicates the amount of cytoplasmic expression of p-IRAK1.

77. The method of any one of claims 5 to 76, wherein the sample expression level of FLT3 indicates the amount of cytoplasmic expression of FLT3.

78. The method of any one of claims 5 to 77, wherein the reference expression level of FLT3 indicates the cytoplasmic expression amount of FLT3.

79. The method of any one of claims 5 to 78, wherein the sample expression level of FLT3 is indicative of the amount of membrane expression of FLT3.

80. The method of any one of claims 5 to 79, wherein the reference expression level of FLT3 indicates the amount of membrane expression of FLT3.

81. The method of any one of claims 5 to 80, wherein the sample expression level of MYD88 indicates the amount of nuclear expression of MYD88.

82. The method of any one of claims 5 to 81, wherein the reference expression level of MYD88 indicates the amount of nuclear expression of MYD88.

83. The method of any one of claims 5 to 82, wherein the sample expression level of MYD88 indicates the amount of cytoplasmic expression of MYD88.

84. The method of any one of claims 5 to 83, wherein the reference expression level of MYD88 indicates the cytoplasmic expression amount of MYD88.

85. The method of any one of claims 5 to 84, wherein the reference level is a value obtained from a subject or subjects not suffering from the disease or disorder.

86. The method of claim 85, wherein the value is obtained from the same biological source as the biological sample (e.g., tissue, blood, or other body fluid).

87. The method of claim 85 or 86, wherein the value is obtained from tissue or blood.

88. The method of any one of claims 85 to 91, wherein the value is obtained from blood.

89. The method of any one of claims 1 to 4, 9 to 12, wherein the cell nucleus is a cancer cell nucleus.

90. The method of any one of claims 1 to 89, wherein the IRAK4 modifying compound is an IRAK4 inhibitor.

91. The method of any one of claims 1 to 89, wherein the IRAK4 modifying compound is an IRAK4 degrader.

92. The method of any one of claims 1 to 91, wherein the IRAK4 inhibitor is represented by Formula I: or a pharmaceutically acceptable salt thereof; in X1 and X3 are independently CH or N; X2 is CR2 or N; wherein one and not more than one of X1, X2 or X3 is N; A is O or S; Y is -CH2- or O; Z is an aryl group or a heterocyclic group; R1 is independently at each occurrence halo or optionally substituted heterocyclyl; wherein the substituent is alkyl, alkoxy, aminoalkyl, halo, hydroxy, hydroxyalkyl or -NR a R b ; R2 is hydrogen, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl or -NR a R b ; wherein the substituent is an alkyl, amino, halide or hydroxyl group; R3, at each occurrence, is alkyl or hydroxy; R a and R b are independently hydrogen, alkyl, acyl or heterocyclyl; 'm' and 'n' are independently 0, 1 or 2; and 'p' is either 0 or 1.

93. The method of claim 92, wherein A is O or S; Y is -CH2- or O; Z is an aryl group or a heterocyclic group; R1 is independently at each occurrence halo or optionally substituted heterocyclyl, wherein the substituent is alkyl, aminoalkyl, halo or -NR a R b ; where R a and R b are independently hydrogen, alkyl or heterocyclyl; R2 is hydrogen, cycloalkyl, heterocyclyl or -NR a R b ; 'm' is 0; and 'n' is 1.

94. The method of claim 92, wherein A is O or S; Y is -CH2- or O; Z is an aryl group or a heterocyclic group; R1 is independently at each occurrence halo or optionally substituted heterocyclyl; wherein the substituent is alkyl, alkoxy, aminoalkyl, halo, hydroxy or -NR a R b ; where R a and R b are independently hydrogen, alkyl or heterocyclyl; R2 is hydrogen, cycloalkyl, optionally substituted heterocyclyl or -NR a R b , wherein the substituent is selected from amino, halo or hydroxy; 'm' and 'n' are independently 0, 1 or 2; and 'p' is either 0 or 1.

95. The method of any one of claims 92 to 94, wherein yes 96. The method of any one of claims 92 to 95, wherein Z is aryl or a 5-membered or 6-membered heterocyclyl.

97. The method of any one of claims 92 to 96, wherein Z is an optionally substituted heterocyclyl selected from the group consisting of phenyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, 1H-tetrazolyl, oxadiazolyl, triazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, azetidinyl, oxetanyl, imidazolidinyl, pyrrolidinyl, oxazolidinyl, thiazolidinyl, pyrazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, 1,4-dioxanyl, dioxythiomorpholinyl, oxapiperazinyl, oxapiperidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, dihydropyranyl and azabicyclo[3.2.1]octyl; each of which is optionally substituted by alkyl, alkoxy, halo, hydroxy, hydroxyalkyl or -NR a R b Replace; and R a and R b are independently hydrogen, alkyl or acyl.

98. The method of claim 92, wherein the IRAK4 inhibitor is represented by formula (IA): or a pharmaceutically acceptable salt thereof.

99. The method of claim 98, wherein A is O or S; Y is -CH2- or O; R1 is independently at each occurrence halo or optionally substituted heterocyclyl, wherein the substituent is alkyl, aminoalkyl, halo or -NR a R b ; where R a and R b are independently hydrogen, alkyl or heterocyclyl; R2 is hydrogen, cycloalkyl, heterocyclyl or -NR a R b ; 'm' is 0; and 'n' is 1.

100. The method of claim 98, wherein A is O or S; Y is -CH2- or O; R1 is independently at each occurrence halo or optionally substituted heterocyclyl; wherein the substituent is alkyl, alkoxy, aminoalkyl, halo, hydroxy or -NR a R b ; where R a and R b are independently hydrogen, alkyl or heterocyclyl; R2 is hydrogen, cycloalkyl, optionally substituted heterocyclyl or -NR a R b , wherein the substituent is selected from amino, halo or hydroxy; and 'm' and 'n' are independently 0, 1 or 2.

101. The method of claim 92, wherein the IRAK4 inhibitor is of formula (IB): or a pharmaceutically acceptable salt thereof.

102. The method of claim 101, wherein A is O or S; Y is -CH2- or O; R1 is independently at each occurrence halo or optionally substituted heterocyclyl; wherein the substituent is alkyl, alkoxy, aminoalkyl, halo, hydroxy or -NR a R b ; where R a and R b are independently hydrogen, alkyl or heterocyclyl; R2 is hydrogen, cycloalkyl, optionally substituted heterocyclyl or -NR a R b , wherein the substituent is selected from amino, halo or hydroxy; and 'm' and 'n' are independently 0, 1 or 2.

103. The method of claim 92, wherein the compound of formula (I) is a compound of formula (IC) or a pharmaceutically acceptable salt thereof.

104. The method of any one of claims 92 to 103, wherein R is an optionally substituted heterocyclyl; wherein the substituent is alkyl, alkoxy, aminoalkyl, halo, hydroxy, hydroxyalkyl, or -NR a R b ; and R a and R b are independently hydrogen or acyl.

105. The method of any one of claims 92 to 103, wherein R is an optionally substituted heterocyclyl; wherein the substituent is alkyl, aminoalkyl, halo or -NR a R b ; and R a and R b are independently hydrogen or acyl.

106. The method of any one of claims 92 to 103, wherein R1 is an optionally substituted heterocyclyl; and the substituent is alkyl, aminoalkyl, halo or -NR a R b ; where R a and R b are independently hydrogen, alkyl or heterocyclyl.

107. The method of any one of claims 92 to 103, wherein R1 is an optionally substituted heterocyclyl; and the substituent is alkyl, alkoxy, aminoalkyl, halo, hydroxyl, or -NR a R b ; where R a and R b are independently hydrogen, alkyl or heterocyclyl.

108. The method of any one of claims 92 to 103, wherein R1 is pyridinyl, pyrazolyl, pyrrolidinyl or piperidinyl.

109. The method of any one of claims 92 to 103, wherein R1 is an optionally substituted pyrazolyl, wherein the substituent is alkyl, hydroxyl or -NR a R b .

110. The method of any one of claims 92 to 103, wherein R1 is halo.

111. The method of any one of claims 92 to 110, wherein R2 is hydrogen, cycloalkyl, heterocyclyl or -NR a R b .

112. The method of any one of claims 92 to 110, wherein R2 is hydrogen, cycloalkyl, optionally substituted heterocyclyl, or -NR a R b , wherein the substituent is selected from amino, halo or hydroxy.

113. The method of any one of claims 92 to 110, wherein R2 is an optionally substituted heterocyclyl selected from piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, azetidinyl, pyrazolyl, furanyl, or azabicyclo[3.2.1]octyl; wherein the substituent is hydroxy, halo, alkyl, or amino.

114. The method of any one of claims 92 to 110, wherein R2 is piperidinyl, pyrrolidinyl, morpholinyl, or piperazinyl.

115. The method of any one of claims 92 to 110, wherein R2 is hydrogen.

116. The method of any one of claims 92 to 110, wherein R2 is cycloalkyl.

117. The method of claim 116, wherein R2 is cyclopropyl.

118. The method of any one of claims 92 to 117, wherein R3 is alkyl.

119. The method of any one of claims 92 to 118, wherein m is 0 and p is 1.

120. The method of any one of claims 92 to 118, wherein m is 0 or 2, and p is 0 or 1.

121. The method of any one of claims 1 to 90, wherein the IRAK4 inhibitor is selected from: or a pharmaceutically acceptable salt or stereoisomer thereof.

122. The method of any one of claims 1 to 91, wherein the IRAK4 inhibitor is 123. The method of any one of claims 1 to 91, wherein the IRAK4 inhibitor is A pharmaceutically acceptable salt of .

124. The method of claim 122 or 123, comprising administering 100-400 mg of the IRAK4 inhibitor to the subject twice daily.

125. The method of claim 122 or 123, comprising administering 200-400 mg of the IRAK4 inhibitor to the subject twice daily.

126. The method of claim 122 or 123, comprising administering 250-350 mg of the IRAK4 inhibitor to the subject twice daily.

127. The method of claim 122 or 123, comprising administering to the subject about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, or about 500 mg of the IRAK4 inhibitor twice daily.

128. The method of claim 122 or 123, comprising administering to the subject about 50 mg, about 75 mg, about 100 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, or about 400 mg of the IRAK4 inhibitor twice daily.

129. The method of claim 122 or 123, comprising administering to the subject about 50 mg, about 100 mg, about 200 mg, or about 300 mg of the IRAK4 inhibitor twice daily.

130. The method of claim 122 or 123, comprising administering to the subject about 200 mg of the IRAK4 inhibitor twice daily.

131. The method of claim 122 or 123, comprising administering to the subject about 225 mg of the IRAK4 inhibitor twice daily.

132. The method of claim 122 or 123, comprising administering to the subject about 250 mg of the IRAK4 inhibitor twice daily.

133. The method of claim 122 or 123, comprising administering to the subject about 275 mg of the IRAK4 inhibitor twice daily.

134. The method of claim 122 or 123, comprising administering to the subject about 300 mg of the IRAK4 inhibitor twice daily.

135. The method of claim 122 or 123, comprising administering to the subject about 325 mg of the IRAK4 inhibitor twice daily.

136. The method of claim 122 or 123, comprising administering to the subject about 350 mg of the IRAK4 inhibitor twice daily.

137. The method of claim 122 or 123, comprising administering to the subject about 375 mg of the IRAK4 inhibitor twice daily.

138. The method of claim 122 or 123, comprising administering to the subject about 400 mg of the IRAK4 inhibitor twice daily.

139. The method of claim 122 or 123, comprising administering to the subject about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, or about 500 mg of the IRAK4 inhibitor once a day.

140. The method of claim 122 or 123, comprising administering to the subject about 50 mg of the IRAK4 inhibitor once daily.

141. The method of claim 122 or 123, comprising administering to the subject about 75 mg of the IRAK4 inhibitor once daily.

142. The method of claim 122 or 123, comprising administering about 100 mg of the IRAK4 inhibitor to the subject once daily.

143. The method of claim 122 or 123, comprising administering to the subject about 125 mg of the IRAK4 inhibitor once daily.

144. The method of claim 122 or 123, comprising administering to the subject about 150 mg of the IRAK4 inhibitor once daily.

145. The method of any one of claims 1 to 44, wherein the IRAK4 inhibitor or IRAK4 degrader is administered orally to the subject.

146. The method of claim 122 or 123, comprising orally administering to the subject about 200 mg of the IRAK4 inhibitor twice daily.

147. The method of claim 122 or 123, comprising orally administering to the subject about 225 mg of the IRAK4 inhibitor twice daily.

148. The method of claim 122 or 123, comprising orally administering to the subject about 250 mg of the IRAK4 inhibitor twice daily.

149. The method of claim 122 or 123, comprising orally administering to the subject about 275 mg of the IRAK4 inhibitor twice daily.

150. The method of claim 122 or 123, comprising orally administering to the subject about 300 mg of the IRAK4 inhibitor twice daily.

151. The method of claim 122 or 123, comprising orally administering to the subject about 325 mg of the IRAK4 inhibitor twice daily.

152. The method of claim 122 or 123, comprising orally administering to the subject about 350 mg of the IRAK4 inhibitor twice daily.

153. The method of claim 122 or 123, comprising orally administering to the subject about 375 mg of the IRAK4 inhibitor twice daily.

154. The method of claim 122 or 123, comprising orally administering to the subject about 400 mg of the IRAK4 inhibitor twice daily.

155. The method of claim 122 or 123, comprising administering to the subject about 50 mg of the IRAK4 inhibitor once daily.

156. The method of claim 122 or 123, comprising administering to the subject about 75 mg of the IRAK4 inhibitor once daily.

157. The method of claim 122 or 123, comprising administering to the subject about 100 mg of the IRAK4 inhibitor once daily.

158. The method of claim 122 or 123, comprising administering to the subject about 125 mg of the IRAK4 inhibitor once daily.

159. The method of claim 122 or 123, comprising administering to the subject about 150 mg of the IRAK4 inhibitor once daily.

160. The method of any one of claims 1 to 91, wherein the IRAK4 modifying compound is PF-06650833 or BAY 1830839.

161. The method of claim 91, wherein the IRAK4 degrader is KT-474.

162. The method of any one of claims 1 to 161, wherein the method further comprises co-administering a BCL-2 inhibitor to the subject.

163. The method of claim 162, wherein the BCL-2 inhibitor is venetoclax.

164. The method of claim 162, comprising administering 400 mg of venetoclax per day.

165. The method of any one of claims 163, wherein the venetoclax is administered orally.

166. The method of claim 163, comprising orally administering 400 mg of venetoclax daily.

167. The method of any one of claims 1 to 161, wherein the method further comprises co-administering a BTK inhibitor to the subject.

168. The method of claim 167, wherein the BTK inhibitor is ibrutinib, acalabrutinib, zanubrutinib, ivotinib, ONO-4059, sipetinib, or HM71224.

169. The method of claim 167, wherein the BTK inhibitor is acalabrutinib.

170. The method of claim 169, comprising administering 200 mg of acalabrutinib per day.

171. The method of claim 169, wherein the acalabrutinib is administered orally.

172. The method of claim 169, comprising orally administering 200 mg of acalabrutinib daily.

173. The method of claim 167, wherein the BTK inhibitor is ibrutinib.

174. The method of claim 173, comprising administering 420 mg of ibrutinib per day.

175. The method of claim 173, comprising administering 560 mg of ibrutinib per day.

176. The method of claim 173, wherein the ibrutinib is administered orally.

177. The method of claim 173, comprising orally administering 420 mg of ibrutinib per day.

178. The method of claim 173, comprising orally administering 560 mg of ibrutinib per day.

179. The method of claim 169, wherein the BTK inhibitor is zanubrutinib.

180. The method of claim 179, comprising administering 160 mg of zanubrutinib twice daily.

181. The method of claim 179, comprising administering 320 mg of zanubrutinib once daily.

182. The method of claim 179, wherein the zanubrutinib is administered orally.

183. The method of claim 179, comprising orally administering 160 mg of zanubrutinib twice daily.

184. The method of claim 179, comprising orally administering 320 mg of zanubrutinib once daily.

185. The method of any one of claims 1 to 184, wherein the disease or disorder is cancer.

186. The method of claim 185, wherein the disease or condition is a hematological malignancy.

187. The method of claim 186, wherein the hematological malignancy is non-Hodgkin's lymphoma.

188. The method of claim 186, wherein the hematological malignancy is a leukemia or a lymphoma.

189. The method of any one of claims 186 to 188, wherein the hematological malignancy is a myeloid leukemia, a myeloid leukemia (e.g., acute myeloid leukemia), a myelodysplastic syndrome, a lymphoblastic leukemia (e.g., acute lymphoblastic leukemia), a chronic lymphoblastic leukemia (CLL), a small lymphocytic lymphoma (SLL), a high-risk CLL, a follicular lymphoma, a diffuse large B-cell lymphoma (DLBCL) (e.g., DLBCL or ABC-DLBLC), a mantle cell lymphoma (MCL), Waldenstrom's macroglobulinemia (WM), multiple myeloma, a marginal zone lymphoma (MZL), a Burkitt's lymphoma, a non-Burkitt's high-grade B-cell lymphoma, an extranodal marginal zone B-cell lymphoma, a transformed high-grade B-cell lymphoma (HGBL), a lymphoplasmacytic lymphoma (LPL), a central nervous system lymphoma (CNSL), or a MALT lymphoma.

190. The method of claim 186, wherein the hematological malignancy is myeloid leukemia.

191. The method of claim 186, wherein the hematological malignancy is a myeloid leukemia (eg, acute myeloid leukemia).

192. The method of claim 186, wherein the hematological malignancy is acute myeloid leukemia (eg, AML).

193. The method of claim 192, wherein the AML is primary AML.

194. The method of claim 192, wherein the AML is secondary AML.

195. The method of any one of claims 192 to 194, wherein the AML is therapy-related AML.

196. The method of claim 186, wherein the hematological malignancy is myelodysplastic syndrome.

197. The method of claim 196, wherein the myelodysplastic syndrome is high grade.

198. The method of claim 196, wherein the myelodysplastic syndrome is low grade.

199. The method of any one of claims 196 to 198, wherein the myelodysplastic syndrome is high risk.

200. The method of claim 186, wherein the hematological malignancy is lymphoblastic leukemia (eg, acute lymphoblastic leukemia).

201. The method of claim 186, wherein the hematological malignancy is chronic lymphoblastic leukemia (CLL).

202. The method of claim 201, wherein the CLL is high-risk CLL.

203. The method of claim 186, wherein the hematological malignancy is small lymphocytic lymphoma (SLL).

204. The method of claim 186, wherein the hematological malignancy is follicular lymphoma.

205. The method of claim 186, wherein the hematological malignancy is diffuse large B-cell lymphoma (DLBCL).

206. The method of claim 186, wherein the hematological malignancy is activated B-cell-like (ABC) DLBCL.

207. The method of claim 186, wherein the hematological malignancy is germinal center B-cell-like (GCB) DLBCL.

208. The method of any one of claims 205 to 207, wherein the DLBCL is extranodal.

209. The method of any one of claims 205 to 208, wherein the DLBCL is extranodal leg lymphoma, extranodal testicular lymphoma, or extranodal not otherwise specified (NOS) lymphoma.

210. The method of claim 186, wherein the hematological malignancy is mantle cell lymphoma.

211. The method of claim 186, wherein the hematological malignancy is Waldenstrom's macroglobulinemia.

212. The method of claim 186, wherein the hematological malignancy is multiple myeloma.

213. The method of claim 186, wherein the hematological malignancy is marginal zone lymphoma.

214. The method of claim 186, wherein the hematological malignancy is Burkitt's lymphoma.

215. The method of claim 186, wherein the hematological malignancy is non-Burkitt's high-grade B-cell lymphoma.

216. The method of claim 186, wherein the hematological malignancy is extranodal marginal zone B-cell lymphoma.

217. The method of claim 186, wherein the hematological malignancy is transformed high-grade B-cell lymphoma (HGBL).

218. The method of claim 186, wherein the hematological malignancy is lymphoplasmacytic lymphoma (LPL).

219. The method of claim 186, wherein the hematological malignancy is a CNS lymphoma.

220. The method of claim 219, wherein the CNS lymphoma is primary CNS lymphoma (PCNSL).

221. The method of claim 186, wherein the hematological malignancy is MALT lymphoma.

222. The method of any one of claims 186 to 221, wherein the hematological malignancy is relapsed.

223. The method of any one of claims 186 to 222, wherein the hematological malignancy is refractory.

224. The method of claim 185, wherein the cancer is selected from brain cancer, kidney cancer, liver cancer, stomach cancer, penile cancer, vaginal cancer, ovarian cancer, gastric cancer, breast cancer, bladder cancer, colon cancer, prostate cancer, pancreatic cancer, lung cancer, cervical cancer, epidermal cancer, prostate cancer, head and neck cancer.

225. The method of claim 185, wherein the cancer is pancreatic cancer.

226. The method of claim 185, wherein the cancer is colon cancer.

227. The method of any one of claims 224 to 226, wherein the cancer is a solid tumor.

228. The method of any one of claims 224 to 227, wherein the cancer is recurrent.

229. The method of any one of claims 224 to 228, wherein the cancer is refractory.

230. The method of any one of claims 1 to 229, wherein the disease or condition is resistant to treatment with a BTK inhibitor.

231. The method of claim 230, wherein the disease or condition is resistant to treatment with ibrutinib, acalabrutinib, zanubrutinib, ivotinib, ONO-4059, sepretinib, or HM71224.

232. The method of claim 230, wherein the disease or condition is resistant to treatment with ibrutinib.

233. The method of claim 230, wherein the disease or condition is resistant to treatment with acalabrutinib.

234. The method of any one of claims 1 to 184, wherein the disease or disorder is an inflammatory disease or disorder.

235. The method of claim 234, wherein the inflammatory disease or disorder is an autoimmune disease or disorder.

236. The method of claim 234, wherein the inflammatory disease or disorder is ocular allergy, conjunctivitis, keratoconjunctivitis sicca, vernal conjunctivitis, allergic rhinitis, an autoimmune blood disorder, hemolytic anemia, aplastic anemia, pure red cell anemia, idiopathic thrombocytopenia, systemic lupus erythematosus,Rheumatoid arthritis, polychondritis, scleroderma, Wegener's granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, Stevens-Johnson syndrome, idiopathic sprue, autoimmune inflammatory bowel disease, ulcerative colitis, Crohn's disease, irritable bowel syndrome, celiac disease, periodontitis, hyaline membrane disease, kidney disease, glomerular disease, alcoholic liver disease, multiple sclerosis, endocrine eye disease, Grave's disease, sarcoidosis, alveolitis, chronic hypersensitivity pneumonitis, primary biliary cirrhosis, uveitis (anterior or posterior), Sjögren's syndrome, interstitial pulmonary fibrosis, psoriatic arthritis, systemic juvenile idiopathic arthritis, nephritis, vasculitis, diverticulitis, interstitial cystitis, glomerulonephritis, idiopathic kidney disease syndrome, minimal change disease, chronic granulomatous disease, endometriosis, leptospirosis nephropathy, glaucoma, retinal disease, headache, pain, complex regional pain syndrome, cardiomegaly, muscle atrophy, metabolic disorders, obesity, fetal growth retardation, hypercholesterolemia, heart disease, chronic heart failure, mesothelioma, anhidrotic urticaria dysplasia, Behcet's disease, pigment disorder, Paget's disease, pancreatitis, hereditary periodic fever syndrome, asthma, acute lung injury, acute respiratory distress syndrome, eosinophilia, hypersensitivity, allergic reaction, fibrositis, gastritis, gastroenteritis, sinusitis, eye allergy, silica-induced disease, chronic obstructive pulmonary disease (COPD), cystic fibrosis, acid Induced lung damage, pulmonary hypertension, polyneuropathy, cataracts, muscle inflammation and systemic sclerosis, inclusion body myositis, myasthenia gravis, thyroiditis, Addison's disease, lichen planus, appendicitis, atopic dermatitis, asthma, allergies, blepharitis, bronchiolitis, bronchitis, bursitis, cervicitis, cholangitis, cholecystitis, chronic transplant rejection, colitis, conjunctivitis, cystitis, dacryoadenitis, dermatitis, juvenile rheumatoid arthritis, dermatomyositis, encephalitis, endocarditis, endometritis, enteritis, enterocolitis, epicondylitis, epididymitis, fasciitis, allergic purpura, hepatitis, suppurative hidradenitis, immunoglobulin A nephropathy, interstitial lung disease, laryngitis, mastitis, meningitis, myelitis, myocarditis, myositis, nephritis, oophoritis , orchitis, osteitis, otitis, pancreatitis, mumps, pericarditis, peritonitis, pharyngitis, urticaria, phlebitis, localized pneumonia, pneumonia, polymyositis, proctitis, prostatitis, pyelonephritis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis, tendinitis, tonsillitis, ulcerative colitis, vasculitis, vulvitis, alopecia areata, erythema multiforme, dermatitis herpetiformis, scleroderma, vitiligo, allergic vasculitis, urticaria, bullous pemphigoid, pemphigus vulgaris, pemphigus foliaceus, paraneoplastic pemphigus, epidermolysis bullosa acquired, acute or chronic gout, chronic gouty arthritis, psoriasis, psoriatic arthritis, rheumatoid arthritis, cryptopyrin-associated periodic syndrome (CAPS), and osteoarthritis. , 237. The method of claim 234, wherein the inflammatory disease or disorder is hypercytokinemia.

238. The method of claim 237, wherein the hypercytokinemia is caused by an infectious agent.

239. The method of claim 238, wherein the infectious agent is a virus.

240. The method of claim 239, wherein the virus is a coronavirus (e.g., COVID-19).

241. The method of claim 238, wherein the infectious agent is a bacterium.

242. The method of claim 234, wherein the inflammatory disease or disorder is graft versus host disease (GVHD).

243. The method of claim 234, wherein the GVHD is chronic graft-versus-host disease (cGVHD).

244. The method of claim 234, wherein the GVHD is scleroderma-like GVHD, steroid-resistant GVHD, cyclosporine-resistant GVHD, GVHD, oral GVHD, reticular oral GVHD, erosive GVHD, or ulcerative oral GVHD.

245. The method of claim 243 or 244, wherein the GVHD is scleroderma-like GVHD.

246. The method of claim 243 or 244, wherein the GVHD is oral GVHD.

247. The method of claim 243 or 244, wherein the GVHD is reticular oral GVHD.

248. The method of claim 243 or 244, wherein the GVHD is erosive GVHD.

249. The method of claim 243 or 244, wherein the GVHD is ulcerative oral GVHD.

250. The method of any one of claims 243 to 249, wherein the GVHD is overlapping chronic GVHD.

251. The method of any one of claims 243 to 249, wherein the GVHD is classical chronic GVHD.

252. The method of any one of claims 243 to 251, wherein the GVHD is steroid-resistant GVHD.

253. The method of any one of claims 243 to 251, wherein the GVHD is cyclosporine-resistant GVHD.

254. The method of any one of claims 243 to 253, wherein the GVHD is refractory.

255. The method of any one of claims 243 to 253, wherein the GVHD is relapsed.

256. The method of any one of claims 1 to 255, wherein the disease or disorder is associated with chronic anemia.

257. The method of any one of claims 1 to 184, wherein the disease or disorder is chronic anemia.

258. The method of any one of claims 1 to 257, wherein the disease or condition is associated with transfusion dependence.

259. The method of any one of claims 1 to 258, wherein the subject is an adult.

260. The method of any one of claims 1 to 259, wherein the subject has previously received at least one anti-cancer therapy (e.g., an anti-cancer therapy or an anti-inflammatory therapy).

261. The method of claim 260, wherein the subject has previously received an anti-cancer therapy.

262. The method of claim 260, wherein the subject has previously received two anti-cancer therapies.

263. The method of claim 260, wherein the subject has previously received three anti-cancer therapies.

264. The method of claim 260, wherein the subject has previously received four anti-cancer therapies.

265. The method of claim 260, wherein the subject has previously received five anti-cancer therapies.

266. The method of any one of claims 260 to 265, wherein the at least one anti-cancer therapy is selected from anti-CD20 antibodies, nitrogen mustards, steroids, purine analogs, DNA topoisomerase inhibitors, DNA intercalators, tubulin inhibitors, BCL-2 inhibitors, proteasome inhibitors, toll-like receptor inhibitors, kinase inhibitors, SRC kinase inhibitors, PI3K kinase inhibitors, BTK inhibitors, glutaminase inhibitors, steroids, PD-1 inhibitors, PD-L1 inhibitors, and methylating agents; or a combination thereof.

267. The method of any one of claims 260 to 266, wherein the anticancer therapy is selected from ibrutinib, rituximab, bendamustine, bortezomib, dexamethasone, chlorambucil, cladribine, cyclophosphamide, doxorubicin, vincristine, venetoclax, ifosfamide, prednisone, oprozomib, ixazomib, acalabrutinib, zanubrutinib, IMO-08400, idelalisib, erbulisel, CB-839, fludarabine, and thalidomide; or a combination thereof.

268. The method of any one of claims 260 to 267, wherein the therapy is dexamethasone.

269. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is ibrutinib.

270. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is ibrutinib and rituximab.

271. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is bendamustine.

272. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is bendamustine and rituximab.

273. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is bortezomib.

274. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is bortezomib and dexamethasone.

275. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is bortezomib and rituximab.

276. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is bortezomib, rituximab, and dexamethasone.

277. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is chlorambucil.

278. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is cladribine.

279. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is cladribine and rituximab.

280. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is cyclophosphamide, doxorubicin, vincristine, prednisone, and rituximab (ie, CHOP-R).

281. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is cyclophosphamide, prednisone, and rituximab (ie, CPR).

282. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is fludarabine.

283. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is fludarabine and rituximab.

284. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is fludarabine, cyclophosphamide, and rituximab.

285. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is rituximab.

286. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is rituximab, cyclophosphamide, and dexamethasone (ie, RCD).

287. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is thalidomide.

288. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is thalidomide and rituximab.

289. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is venetoclax.

290. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is cyclophosphamide, bortezomib, and dexamethasone (ie, R-CyBorD).

291. The method of any one of claims 260 to 267, wherein the anti-cancer therapy is a hypomethylating agent.

292. The method of any one of claims 1 to 291, wherein the subject has previously received at least 6 cycles of a hypomethylating agent.

293. The method of any one of claims 1 to 292, wherein the subject has previously received etoposide chemomobilization therapy.

294. The method of any one of claims 1 to 293, wherein the subject has previously received a bone marrow transplant.

295. The method of any one of claims 1 to 294, wherein the subject has previously received a hematopoietic cell transplant.

296. The method of any one of claims 1 to 295, wherein the subject has previously received a stem cell transplant.

297. The method of any one of claims 1 to 296, wherein the subject has previously received an autologous stem cell transplant.

298. The method of any one of claims 1 to 297, wherein the subject has previously received an allogeneic stem cell transplant.

299. The method of any one of claims 1 to 298, wherein the subject has previously received carmustine, etoposide, cytarabine, and melphalan (i.e., BEAM conditioning).

300. The method of any one of claims 1 to 299, wherein the subject has previously received re-induction therapy.

301. The method of any one of claims 1 to 300, wherein the subject has previously achieved a partial response.

302. The method of any one of claims 1 to 301, wherein the subject has previously achieved a good partial response.

303. The method of any one of claims 1 to 302, wherein the subject has previously achieved a complete response.

304. The method of any one of claims 1 to 303, wherein the subject has a mutation in RICTOR.

305. The method of any one of claims 1 to 304, wherein the subject has an N1065S mutation in RICTOR.

306. The method of any one of claims 1 to 305, wherein the subject has a mutation in MYD88.

307. The method of any one of claims 1 to 306, wherein the subject has a L265P mutation in MYD88.

308. The method of any one of claims 1 to 307, wherein the subject has a mutation in TET2.

309. The method of any one of claims 1 to 308, wherein the subject does not have a mutation in CXCR4.

310. The method of any one of claims 1 to 308, wherein the subject has a mutation in CXCR4.

311. The method of any one of claims 1 to 310, wherein the subject exhibits early progression.

312. The method of any one of claims 1 to 311, wherein the subject has not previously received a BTK inhibitor.

313. The method of any one of claims 1 to 312, wherein following administration of the IRAK4 inhibitor, the subject achieves a partial response.

314. The method of any one of claims 1 to 312, wherein following administration of the IRAK4 inhibitor, the subject achieves a good partial response.

315. The method of any one of claims 1 to 312, wherein following administration of the IRAK4 inhibitor, the subject achieves a complete response.

316. The method of any one of claims 1 to 315, wherein following administration of the IRAK4 inhibitor, the subject has reduced IL-1 induced signaling.

317. The method of any one of claims 1 to 316, wherein following administration of the IRAK4 inhibitor, the subject's cytokine production is reduced.

318. The method of any one of claims 1 to 317, wherein the IRAK4 inhibitor is administered until disease progression or unacceptable toxicity.

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