TW201427666A - Methods of treating a disease or disorder associated with Bruton's tyrosine kinase - Google Patents

Abstract

The present invention provides methods of treating, stabilizing or lessening the severity or progression of a disease or disorder associated with BTK.

Description

Method for treating diseases or disorders associated with BRUTON's tyrosine kinase

Cross-reference to related applications

This application claims US Provisional Application No. 61/728,701, filed on November 20, 2012, No. 61/728,703, filed on Nov. 20, 2012, and No. 61/799,761, filed on March 15, 2013, Priority No. 61/870,718, filed on August 27, 2013, and No. 61/870,720, filed on Aug. 27, 2013, each of which is incorporated herein by reference in its entirety. .

The present invention provides methods of treating, stabilizing or ameliorating the severity or progression of a disease or disorder associated with Bruton's Tyrosine Kinase (BTK).

In recent years, the search for novel therapeutic agents has been greatly assisted by a better understanding of the structure of enzymes and other biomolecules associated with diseases. One of the most important enzymes that have become a widely studied subject is protein kinases.

Protein kinases constitute a large family of structurally related enzymes responsible for controlling multiple signal transduction processes within cells. Since the structure and catalytic function of protein kinases are conserved, they are thought to evolve from common ancestral genes. Almost all kinases contain a similar catalytic domain of 250-300 amino acids. Kinases can be classified into families according to their phosphorylation (eg, protein-tyrosine, protein-serine/threonine, lipid, etc.).

In general, protein kinases mediate intracellular signaling by affecting the phosphoryl transfer from the nucleoside triphosphate to the protein receptor involved in the signaling pathway. These phosphorylation events act as molecular on/off transducers that modulate or regulate the biological function of the target protein. Responses to a variety of extracellular stimuli and other stimuli ultimately trigger these phosphorylation events. Examples of such stimuli include environmental and chemical stress signals (eg, osmotic shock, thermal shock, ultraviolet radiation, bacterial endotoxin and H ₂ O ₂ ), cytokines (eg, interleukin-1 (IL-1), and tumor necrosis). Factor alpha (TNF-alpha) and growth factors (eg, granule macrophage colony stimulating factor (GM-CSF) and fibroblast growth factor (FGF)). Extracellular stimulation can affect one or more cellular responses associated with cell growth, migration, differentiation, hormone secretion, activation of transcription factors, glucose metabolism, control of protein synthesis, and regulation of the cell cycle.

A variety of diseases are associated with abnormal cellular responses elicited by events mediated by protein kinases as described above. Such diseases include, but are not limited to, autoimmune diseases, inflammatory diseases, bone diseases, metabolic diseases, neurodegenerative diseases, cancer, cardiovascular diseases, allergies and asthma, Alzheimer's disease (Alzheimer's disease) ) and hormone-related diseases. Therefore, there is still a need to find protein kinase inhibitors that are useful as therapeutic agents.

Chronic lymphocytic leukemia (CLL) is a lymphoid proliferative malignancy characterized by progressive accumulation of morphologically mature but functionally incomplete lymphocytes in blood, bone marrow, and lymphoid tissues. It mainly affects older individuals with a median age of 65 to 70 years. Small lymphocytic lymphoma (SLL) and CLL are generally considered to be different manifestations of the same disease. CLL is found in the blood and bone marrow, while SLL is mainly found in lymph nodes. The clinical course of CLL/SLL is between invasive disease with a long-term survival of more than 12 years and a median survival of 2 years. The mean age of diagnosis with CLL/SLL is approximately 60 years.

Despite the recent approval of therapeutics and combination therapies, CLL/SLL remains incurable and most patients eventually relapse and/or die. Improved novel combination therapies for individuals with CLL/SLL requiring treatment still do not meet medical needs.

Bruton tyrosine kinase (Btk) is a non-receptor tyrosine kinase that is primarily restricted to B lymphocytes, monocytes, and restricted cell expression of mast cells or basophils. Btk is a key component of the B cell receptor (BCR) signaling network and is critical for B cell development. Investigations have shown that some B cell malignancies (including diseases such as CLL/SLL) rely on BCR signaling, suggesting that disrupting this signaling can be a promising therapeutic opportunity. Recently, agents that inhibit spleen tyrosine kinase (Syk) and Btk (i.e., two components of the BCR signaling pathway) have been reported for various B-cell non-Hodgkin's Lymphoma (NHL). And clinical anti-tumor response in CLL/SLL.

U.S. Published Patent Application No. US 2010/0029610, the entire disclosure of which is hereby incorporated by reference in its entirety in its entirety in the entire entire entire entire entire entire entire entire entire entire disclosure Certain 2,4-disubstituted pyrimidine compounds that are active in kinases, including BTK, a member of the TEC kinase. Such compounds include N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilinyl)pyrimidin-4-ylamino)phenyl)propene, hereinafter referred to as Compound 1 . Indoleamine, which is named Compound No. I-182 in the '610 publication. The synthesis of Compound 1 is described in detail in Example 20 of the '610 publication. Compound 1 is active in a variety of assays and therapeutic models, demonstrating covalent and irreversible inhibition of BTK (in enzymatic and cellular assays). Notably, Compound 1 is an effective, selective, orally administrable small molecule that has been found to inhibit B cell proliferation and activation. Compound 1 is therefore suitable for the treatment of one or more disorders associated with BTK activity.

Accordingly, the present invention provides, inter alia, methods of treating, stabilizing or ameliorating the severity or progression of one or more diseases and conditions associated with BTK. In some aspects, the invention provides methods of treating, stabilizing or ameliorating the severity or progression of one or more BTK-related diseases and conditions, comprising administering to a patient in need thereof a pharmaceutically acceptable composition, The composition comprises N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl)propenylamine ( 1 ):

In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of one or more BTK-related diseases and conditions, comprising administering Compound 1 with lenalidomide to a patient in need thereof ( The combination of lenalidomide).

In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of one or more BTK-associated diseases and conditions, comprising administering to a patient in need thereof a composition comprising Compound 1 and comprising A combination of lenalidomide compositions.

In some embodiments, the methods provided comprise administering a combination of Compound 1 and lenalidomide to a patient in need thereof, wherein Compound 1 is administered once a day. In some embodiments, the methods provided comprise administering a combination of Compound 1 and lenalidomide to a patient in need thereof, wherein Compound 1 is administered twice a day. In some of these embodiments, lenalidomide is administered once a day. Thus, in some embodiments, the methods provided comprise administering a combination of Compound 1 and lenalidomide to a patient in need thereof, wherein Compound 1 is administered twice a day and lenalidomide is administered once a day.

In some embodiments, a method is provided comprising administering to a patient in need thereof a composition comprising Compound 1 and lenalidomide.

In some embodiments, the disease or condition associated with BTK is selected from the group consisting of chronic lymphocytic leukemia and small lymphocytic lymphoma.

In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of chronic lymphocytic leukemia (CLL) comprising administering a combination of Compound 1 and lenalidomide to a patient in need thereof .

In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of small lymphoplastic lymphoma (SLL) comprising administering Compound 1 with lenalidomide to a patient in need thereof combination.

In some embodiments, the therapy provided comprises orally administering to a patient a combination of Compound 1 and lenalidomide. In some embodiments, Compound 1 and lenalidomide are each administered as a pharmaceutical formulation. In some of these embodiments, the formulations are capsule formulations.

In some embodiments, the invention also provides a dosing regimen and schedule for administering a combination of Compound 1 and lenalidomide to a patient in need thereof.

In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of one or more BTK-related diseases and conditions, comprising administering Compound 1 , lenalidomide, and, to a patient in need thereof Each of the anti-CD20 antibodies (eg, rituximab). In some such embodiments, the disease or condition associated with BTK is selected from the group consisting of chronic lymphocytic leukemia and small lymphocytic lymphoma.

The methods, dosing regimens and plans for administration of the combination are described in further detail below.

Figure 1 presents a specific dose response curve for the combination of Compound 1 besylate and lenalidomide (3000 nM) in the OCI-LY-10 cell line. • = compound 1 besylate; □ = lenalidomide; Δ = calculated or expected combined activity; ◇ = observed combined activity. Compound 1 besylate and lenalidomide appeared to exhibit an additive effect in the OCI-LY-10 cell line.

Figure 2 presents a "volcano" plot of the combination of Compound 1 besylate (0.2 nM - 1500 nM) and lenalidomide (300 nM, 1000 nM and 3000 nM) in the OCI-LY-10 cell line. Compound 1 besylate and lenalidomide appeared to exhibit an additive effect in the OCI-LY-10 cell line.

Figure 3 presents a specific dose response curve for the combination of Compound 1 besylate and lenalidomide (3333 nM) in the WSU-DLCL2 cell line. • = compound 1 besylate; □ = lenalidomide; Δ = calculated or expected combined activity; ◇ = observed combined activity. Compound 1 besylate and lenalidomide appeared to exhibit an additive effect in the WSU-DLCL2 cell line.

Figure 4 presents a "volcano" plot of the combination of Compound 1 besylate (0.5 nM - 3333 nM) and lenalidomide (123 nM, 3333 nM and 10000 nM) in the WSU-DLCL2 cell line. Compound 1 and benzenesulfonate lenalidomide seems to show additive effects in cell line WSU-DLCL2.

5 presents certain cell lines Dose-response curves of Compound 1 Riva benzenesulfonate and lenalidomide (3333nM) of the combination. • = compound 1 besylate; □ = lenalidomide; Δ = calculated or expected combined activity; ◇ = observed combined activity. Significant synergies are indicated by arrows.

Figure 6 presents a "volcano" plot of the combination of Compound 1 besylate (0.2 nM - 1000 nM) and lenalidomide (41 nM, 1111 nM and 3333 nM) in the Riva cell line. Significant synergies are indicated by arrows.

Figure 7 presents a specific dose response curve for the combination of Compound 1 besylate and lenalidomide (333 nM) in the Riva cell line. • = compound 1 besylate; □ = lenalidomide; Δ = calculated or expected combined activity; ◇ = observed combined activity. Significant synergies are indicated by arrows.

Figure 8 presents a "volcano" plot of the combination of Compound 1 besylate (0.2 nM - 1000 nM) and lenalidomide (41 nM, 123 nM and 370 nM) in the Riva cell line. Significant synergies are indicated by arrows.

Figure 9 presents a specific dose response curve for the combination of Compound 1 besylate and lenalidomide (333 nM) in the Riva cell line. • = compound 1 besylate; □ = lenalidomide; Δ = calculated or expected combined activity; ◇ = observed combined activity.

Figure 10 presents a "volcano" plot of the combination of Compound 1 besylate (0.2 nM - 1000 nM) and lenalidomide (41 nM, 123 nM and 370 nM) in the Riva cell line. Compound 1 besylate and lenalidomide appeared to exhibit an additive effect in the Riva cell line.

Figure 11 presents a specific dose response curve for the combination of Compound 1 besylate and lenalidomide (3000 nM) in the TMD-8 cell line. • = compound 1 besylate; □ = lenalidomide; Δ = calculated or expected combined activity; ◇ = observed combined activity. Significant synergies are indicated by arrows.

Figure 12 presents a "volcano" plot of the combination of Compound 1 besylate (0.2 nM - 1500 nM) and lenalidomide (300 nM, 1000 nM and 3000 nM) in the TMD-8 cell line. Significant synergies are indicated by arrows.

Figure 13 presents a response assessment of patients enrolled in Group 1 up to October 16, 2013.

Figure 14 presents the reaction assessment to date October 16, 2013 Group 2 patients registered.

definition

As used herein, the terms "combination", "in combination with" or "combination therapy" mean that two or more different pharmaceutical agents are administered in an overlapping regimen such that the individual is simultaneously exposed to both agents. The situation. In some embodiments, the combinations are directed to the individual dosage form of Compound 1 and lenalidomide administered simultaneously. In some embodiments, the combinations are directed to an individual to simultaneously administer a separate dosage form of Compound 1 and lenalidomide, wherein Compound 1 is administered before, during, or after administration of lenalidomide. In some embodiments, simultaneous or simultaneous exposure of Compound 1 and lenalidomide is achieved via different dosing regimens appropriate to each therapeutic agent. For example, in some embodiments, Compound 1 is administered twice daily or BID, while lenalidomide is administered once daily or QD. Alternatively and/or additionally, Compound 1 may be administered once or twice daily for one or more 28-day cycles, while lenalidomide may be administered once daily for one or more of the first 28 days of the 28-day cycle. Day to the 21st day. In some embodiments, Compound 1 is administered twice daily from day 8 to day 28 of one or more 28 day cycles, and lenalidomide is on day 1 to day of one or more 28 day cycles. Once a day for 28 days. In some embodiments, Compound 1 is administered twice daily from day 1 to day 28 of one or more 28 day cycles, and lenalidomide is on day 1 to day of one or more 28 day cycles. Once a day for 28 days.

The term "percent inhibition" as used herein refers to a test compound (eg, no Percentage of target activity relative to control target activity decreased in the presence of a reversible BTK inhibitor. It will be appreciated that the percent inhibition of a target (e.g., kinase) can be determined in a number of ways, one of which is described in Example 3 below. In some embodiments, the percent inhibition is expressed as % inhibition (eg, 50% inhibition). In some embodiments, the percent inhibition of the kinase is the average percent inhibition.

As used herein, the term "comparable" means that two or more agents, entities, conditions, conditional settings, etc., may not be identical to each other, but are similar enough to allow comparison therebetween so that they can be based on observations Differences or similarities are reasonably drawn. Those of ordinary skill in the art will appreciate that, in this context, the degree of consistency required for two or more such agents, entities, conditions, conditional settings, etc., in any given environment will be considered comparable. . As used herein, the terms " percent of comparable inhibition " or " percentage of comparable mean inhibition " refer to a kinase within 10% of the percent inhibition or percent inhibition of a reference kinase inhibitor observed or measured, respectively. Percent inhibition or average percent inhibition. For example, if the reference kinase inhibitor has 50% inhibition of the kinase relative to the control and the other inhibitor has about 40% to about 60% inhibition of the same kinase relative to the control, the other inhibitor should be considered as exhibitable. Comparison of inhibition. In some embodiments, the irreversible BTK inhibitor has a percent inhibition comparable to a reference kinase inhibitor, wherein the percent inhibition of the irreversible BTK inhibitor is 9%, 8 percent of the percent inhibition of the reference kinase inhibitor observed or determined. %, 7%, 6%, 5%, 4%, 3%, 2% or 1% inhibition.

As used herein, "a disease or disorder associated with BTK" or "a BTK-mediated disorder" means any disease or other deleterious condition in which BTK or a mutant thereof is known or suspected to function. Accordingly, another embodiment of the invention relates to the prevention, treatment, stabilization or amelioration of the severity or progression of one or more diseases known or suspected to function as BTK or a mutant thereof. In particular, the invention relates to a method of treating or ameliorating the severity of a proliferative disorder, wherein the method comprises administering to a patient in need thereof a combination of Compound 1 and lenalidomide.

As used herein, the term "irreversible" or "irreversible inhibitor" refers to an inhibitor (ie, a compound) that is capable of covalently binding to a protein kinase of interest in a substantially non-reversible manner. That is, in view of the ability of a reversible inhibitor to bind to (but generally fail to form a covalently bound) target protein kinase, and thus dissociate from a protein kinase of interest, the irreversible inhibitor will still substantially bind after covalent binding has occurred. Target protein kinase. Irreversible inhibitors generally show time dependence , whereby the degree of inhibition increases with the time the inhibitor is in contact with the enzyme. Methods for identifying whether a compound acts as an irreversible inhibitor are known to those of ordinary skill in the art. Such methods include, but are not limited to, enzymatic kinetic analysis of inhibition profiles of compounds and protein kinase targets, mass spectrometric analysis using protein drug targets modified in the presence of inhibitor compounds, discontinuous exposure (also known as "elution ( Washout)) experiments, and the use of labels (such as radiolabeled inhibitors) to display covalent modifications of the enzyme, as well as other methods known to those skilled in the art.

The term "refractory CLL/SLL" as used herein is defined as CLL/SLL treated with at least one line of prior therapy (i) not achieving at least partial response to therapy or (ii) progressing within 6 months of treatment.

The term "recurrent CLL/SLL" as used herein is defined as after treatment After 6 months, CLL/SLL progressed after obtaining partial or complete response to the therapy.

The term "individual" as used herein means a mammal and includes both human and animal individuals, such as livestock (eg, horses, dogs, cats, etc.).

As used herein, "therapeutically effective amount" means the amount of a substance (eg, a therapeutic, composition, and/or formulation) that elicits a desired biological response. In some embodiments, a therapeutically effective amount of a substance is sufficient to treat, diagnose, prevent, treat, and/or prevent the disease, disorder, and/or when administered to an individual suffering from or susceptible to a disease, disorder, and/or condition. Or the condition and/or delay the amount of its onset. Those of ordinary skill in the art will appreciate that the effective amount of a substance may vary depending on factors such as the desired biological endpoint, the substance to be delivered, the target cell or tissue, and the like. Chemical. For example, an effective amount of a compound in a formulation to treat a disease, disorder, and/or condition is to alleviate, ameliorate, alleviate, inhibit, prevent, or delay one or more symptoms or characteristics of the disease, disorder, and/or condition. The amount that attacks, reduces its severity, and/or reduces its incidence. In some embodiments, a "therapeutically effective amount" is a compound or a composition comprising a compound that is sufficient to treat at least a minimum amount of one or more symptoms of a disorder or condition associated with Bruton tyrosine kinase.

The term "treating" as used herein refers to partial or complete alleviation, inhibition, delay of onset, prevention, amelioration and/or alleviation of a disorder or condition, or one or more symptoms of the disorder or condition. As used herein, the term "treating" refers to partial or complete alleviation, inhibition, delay of onset, prevention, amelioration and/or alleviation of a disorder or condition, or one or more symptoms of the disorder or condition, as described herein. In some embodiments, the treatment can be administered after one or more symptoms have developed. In some embodiments, the term "treating" includes preventing a disease or disorder or interrupting its progression. In other embodiments, the treatment can be administered in the absence of symptoms. For example, treatment can be administered to a susceptible individual prior to the onset of symptoms (eg, based on a history of symptoms and/or according to genetic or other susceptibility factors). Treatment can also continue after the symptoms have subsided, for example to prevent or delay their reproduction. Thus, in some embodiments, the term "treatment" includes preventing the recurrence or recurrence of a disease or disorder.

The expression "unit dosage form" as used herein refers to a physically discrete unit of a therapeutic formulation suitable for the individual to be treated. It will be understood, however, that the total daily usage of the compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The particular effective amount of any particular individual or organism will depend on a number of factors, including the disorder being treated and the severity of the disorder; the activity of the particular active agent employed; the particular composition employed; the age, weight, and general health of the subject Condition, sex and diet; time of administration, and excretion rate of the particular active agent used; duration of treatment; drugs and/or additional therapies used in combination or superimposed with the particular compound employed, and similar factors well known in the medical arts.

As used herein, the term "antibody" or a variant thereof (ie, an antibody) refers to a polypeptide that is capable of binding to an epitope. In some embodiments, the antibody is a full length antibody. In some embodiments, the antibody is less than the full length (ie, the antibody fragment) but includes at least one binding site. In some such embodiments, the binding site comprises at least one and preferably at least two sequences having an antibody variable region structure. In some embodiments, the term "antibody" encompasses any protein whose binding domain is homologous or largely homologous to an immunoglobulin binding domain. In a specific embodiment, the term "antibody" encompasses a binding domain that displays a polypeptide that is at least 99% identical to an immunoglobulin binding domain. In some embodiments, the antibody exhibits at least 70%, at least 80%, at least 85%, at least 90%, or at least 95% identity to the immunoglobulin binding domain for the binding domain. The antibody polypeptides of the invention can be prepared by any available means, including, for example, isolation from a natural source or antibody library, recombinant production in a host system or using a host system, chemical synthesis, and the like, or a combination thereof. In some embodiments, the antibody is single or multiple. In some embodiments, the antibody can be a member of any of the immunoglobulin classes, including any of the human classes IgG, IgM, IgA, IgD, and IgE. In certain embodiments, the antibody is a member of the IgG immunoglobulin class. In some embodiments, the term "antibody" refers to any derivative of an antibody that has the ability to bind to an epitope of interest. In some embodiments, antibody fragments comprise a plurality of strands that are linked together, for example, by disulfide linkages. In some embodiments, the antibody is a human antibody. In some embodiments, the antibody is a humanized antibody. In some embodiments, the humanized antibody comprises a chimeric immunoglobulin, immunoglobulin chain or antibody fragment (Fv, Fab, Fab', F(ab') ₂ or comprising a minimal sequence derived from a non-human immunoglobulin or Other antigen binding sequence of the antibody). In some embodiments, the humanized antibody is a human immunoglobulin (receptor antibody), wherein the residues of the complementarity determining regions (CDRs) of the receptor are passed through a non-human species having the desired specificity, affinity, and ability (for Subbody antibodies), such as residues of CDRs of mouse, rat or rabbit. In a particular embodiment, the antibody used in the invention binds to a particular epitope of CD20. In some embodiments, the epitope of CD20 that binds to an anti-CD20 antibody includes, for example, ¹⁷⁰ ANPS ¹⁷³ (Binder et al, Blood 2006, 108(6): 1975-1978), FMC7 (Deans et al, Blood 2008, 111). (4): 2492), Rp5-L and Rp15-C (simulated epitope of CD20) (Perosa et al. , J. Immunol. 2009, ¹⁸² : 416-423), ¹⁸² YCYSI ¹⁸⁵ (Binder et al., Blood 2006, 108(6): 1975-1978) and WEWTI (model of ¹⁸² YCYSI ¹⁸⁵ ) (Binder et al., Blood 2006, 108(6): 1975-1978). In some embodiments, the anti-CD20 antibody binding affinity of the epitope of the CD20 antigen (K _d) of less than 12nM, less than 11 nM, less than 10 nM, less than 9nM, less than 8nM, less than 7 nM, less than 6nM, less than of 5 nM, less than 4nM, less than 3nM, less than 2nM or less than 1nM.

As used herein, the term (eg, an approved reference product/biological drug, such as a protein therapeutic, an antibody, etc.) "biological analog" refers to a biological product that is similar to a reference product based on the following information: a) Analytical studies demonstrating that biological products are highly similar to reference products despite minor differences in clinically inactive components; (b) animal studies (including toxicity assessment); and/or (c) clinical studies (including immunogens) Sexual and pharmacodynamic or pharmacodynamic evaluation) sufficient to demonstrate the safety, purity and efficacy of the reference product under approved and intended use and for approval of one or more suitable conditions of use (eg for product safety) In terms of purity and efficacy, there is no clinically meaningful difference between biological products and reference products).

In some embodiments, the biosimilar biological product and the reference product utilize the same mechanism of action for the conditions of use specified, recommended, or proposed in the proposed label, but only if the mechanism of action of the reference product is known. In some embodiments, the conditions of use specified, recommended, or proposed in the suggested label for the biological product have been previously approved for use in the reference product. In some embodiments, the biopharmaceutical route of administration, dosage form, and/or concentration is the same as the reference product. In some embodiments, the facility that manufactures, processes, packages, or holds the biological product conforms to standards designed to ensure that the biological product remains safe, pure, and effective. The reference product is approved in at least one of the United States, Europe, or Japan. The biological analog can be, for example, a currently known antibody having the same amino acid sequence as the commercially available antibody, but can be produced in different cell types or by different methods of production, purification or formulation.

A general method for treating BTK-mediated diseases or disorders

In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of one or more BTK-related diseases and conditions, comprising administering an irreversible BTK inhibitor to a patient in need thereof amine.

It will be understood that although the methods described herein refer to the formulation, dosage and dosing schedule/time course of Compound 1 and its salts, such formulations, dosages and/or dosing schedules/time schedules are equally applicable to any irreversible BTK inhibitors, such as those described below, are irreversible BTK inhibitors. Thus, in some embodiments, the dosage or dosing regimen of the irreversible BTK inhibitor is selected from any of the doses or dosing regimens of Compound 1 as described herein. In some embodiments, the methods provided comprise administering an irreversible BTK inhibitor in an amount selected from any of the doses of Compound 1 as described herein. In some such embodiments, a dose of an irreversible BTK inhibitor is administered according to the time course of administration of any of the dosing schedules selected from Compound 1 described herein. In some embodiments, the composition comprising an irreversible BTK inhibitor is any formulation as described herein.

In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of one or more BTK-related diseases and conditions, comprising administering an irreversible BTK inhibitor to a patient in need thereof, Amines and anti-CD20 antibodies (eg rituximab).

In some embodiments, the irreversible BTK inhibitor is covalently bound to Cys481 of BTK.

In some embodiments, the irreversible BTK inhibitor has activity against one or more kinases selected from the kinases listed in Table 7 below.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition comparable to a reference kinase inhibitor for a kinase selected from Table 7 or a combination thereof. In some such embodiments, the reference kinase inhibitor is Compound 2 :

In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition comparable to Compound 2 for one or more kinases selected from Table 7 or a combination thereof, wherein the percent inhibition of the irreversible kinase inhibitor is observed with respect to Compound 2 Within about 10% of the percent inhibition. In some embodiments, the irreversible BTK inhibitor has a percent inhibition comparable to Compound 2 for one or more kinases selected from Table 7 or a combination thereof, wherein the percent inhibition of the irreversible kinase inhibitor is observed with respect to Compound 2 Up to about 9%, or about 8%, or about 7%, or about 6%, or about 5%, or about 4%, or about 3%, or about 2% or about 1% inhibition of the percent inhibition.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of one or more kinases selected from Table 7 that is greater than the percent inhibition observed for Compound 2 . In some embodiments, the irreversible BTK inhibitor has a percent inhibition of one or more kinases selected from Table 7 that is less than the percent inhibition observed for Compound 2 .

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of one or more additional kinases, wherein the percent inhibition of the one or more kinases is at least about 50%, at least about 55%, at least about 60%, at least about 65%, At least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95%.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition comparable to a reference kinase inhibitor for one or more kinases selected from the group consisting of: TXK, BMX/ETK, FLT3, BLK, TEC, ERBB4/ HER4, Aurora B, TRKC, RET, LOK/STK10, Aurora C, FLT4/VEGFR3, ROS/ROS1, ARK5/NUAK1, EGFR, DDR1, JAK3, LRRK2, ABL2/ARG, ITK, Aurora A, YES/YES1, FGFR3 , TNK1, BRK, FGFR2, PDGFRb, c-SRC, ACK1, FGFR1, STK16, ABL1, AXL, TYK2, ERBB2/HER2, FGR, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2 and CLK2 , or a combination thereof. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percentage of inhibition comparable to a reference kinase inhibitor for a panel of kinases consisting of: TXK, BMX/ETK, FLT3, BLK, TEC, ERBB4/HER4, Aurora B , TRKC, RET, LOK/STK10, Aurora C, FLT4/VEGFR3, ROS/ROS1, ARK5/NUAK1, EGFR, DDR1, JAK3, LRRK2, ABL2/ARG, ITK, Aurora A, YES/YES1, FGFR3, TNK1, BRK , FGFR2, PDGFRb, c-SRC, ACK1, FGFR1, STK16, ABL1, AXL, TYK2, ERBB2/HER2, FGR, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2 and CLK2, or a combination thereof . In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor inhibits a kinase selected from the group consisting of: TXK, BMX/ETK, FLT3, BLK, TEC, ERBB4/HER4, Aurora B, TRKC, RET, LOK/STK10, Aurora C, FLT4/VEGFR3, ROS/ROS1, ARK5/NUAK1, EGFR, DDR1, JAK3, LRRK2, ABL2/ARG, ITK, Aurora A, YES/YES1, FGFR3, TNK1, BRK, FGFR2, PDGFRb, c-SRC, ACK1 , FGFR1, STK16, ABL1, AXL, TYK2, ERBB2/HER2, FGR, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2 and CLK2, or a combination thereof, wherein the inhibition of the one or more kinases The percentage inhibition of at least the reference kinase inhibitor observed. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has at least about 50%, at least about 55%, at least about 60%, at least 65%, at least about 70%, at least about 75%, at least about a kinase selected from the group consisting of: 80%, at least about 85%, at least about 90% or at least about 95% inhibition: TXK, BMX/ETK, FLT3, BLK, TEC, ERBB4/HER4, Aurora B, TRKC, RET, LOK/STK10, Aurora C, FLT4 /VEGFR3, ROS/ROS1, ARK5/NUAK1, EGFR, DDR1, JAK3, LRRK2, ABL2/ARG, ITK, Aurora A, YES/YES1, FGFR3, TNK1, BRK, FGFR2, PDGFRb, c-SRC, ACK1, FGFR1 STK16, ABL1, AXL, TYK2, ERBB2/HER2, FGR, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2 and CLK2, or a combination thereof.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition comparable to a reference kinase inhibitor for one or more kinases selected from the group consisting of: Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1 ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1, STK16, ABL1, AXL, TYK2, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2 and CLK2, or a combination thereof. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition comparable to a reference kinase inhibitor for a panel of kinases consisting of: Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1 , LRRK2, ABL2/ARG, TNK1, STK16, ABL1, AXL, TYK2, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2, and CLK2, or a combination thereof. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the percent inhibition of the irreversible BTK inhibitor against one or more kinases selected from the group consisting of at least the observed percent inhibition of the reference kinase inhibitor: Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1, STK16, ABL1, AXL, TYK2, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2 and CLK2, or combination. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least at least a kinase selected from the group consisting of: About 80%, at least about 85%, at least about 90% or at least about 95% inhibition: Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1, STK16, ABL1 , AXL, TYK2, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2, and CLK2, or a combination thereof.

In some embodiments, the invention provides a method of treating, stabilizing or attenuating the severity or progression of CLL/SLL comprising administering an irreversible BTK inhibitor to a patient in need thereof And lenalidomide, wherein the irreversible BTK inhibitor has no more than about 50% inhibition of a kinase selected from the group consisting of: Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2 /ARG, TNK1, STK16, ABL1, AXL, TYK2, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2, and CLK2, or a combination thereof.

In some embodiments, the irreversible BTK inhibitor has at least about 50% inhibition of a kinase selected from the group consisting of Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1, STK16, ABL1, AXL, TYK2, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, and PKCb2, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has at least about 50% inhibition of a panel of kinases consisting of: Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ ARG, TNK1, STK16, ABL1, AXL, TYK2, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2 and CLK2.

In some embodiments, the irreversible BTK inhibitor has at least about 55% inhibition of a kinase selected from the group consisting of Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1, STK16, ABL1, CHK2, MLK1/MAP3K9, MLK2/MAP3K10 and MLK3/MAP3K11, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has at least about 55% inhibition by one of a group of kinases: Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ ARG, TNK1, STK16, ABL1, CHK2, MLK1/MAP3K9, MLK2/MAP3K10 and MLK3/MAP3K11.

In some embodiments, the irreversible BTK inhibitor has at least about 60% inhibition of a kinase selected from the group consisting of: Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1, STK16, CHK2, MLK1/MAP3K9 and MLK3/MAP3K11, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has at least about 60% inhibition of a panel of kinases consisting of: Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ ARG, TNK1, STK16, CHK2, MLK1/MAP3K9 and MLK3/MAP3K11.

In some embodiments, the irreversible BTK inhibitor has at least about 65% inhibition of a kinase selected from the group consisting of: Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, TNK1, STK16, CHK2, MLK1/MAP3K9 and MLK3/MAP3K11, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has at least about 65% inhibition by one of a group of kinases: Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, TNK1 STK16, CHK2, MLK1/MAP3K9 and MLK3/MAP3K11.

In some embodiments, the irreversible BTK inhibitor has at least about 70% inhibition of a kinase selected from the group consisting of Aurora A, Aurora B, Aurora C, ROS/ROS1, ARK5/NUAK1, TNK1, STK16, CHK2, MLK1/ MAP3K9 and MLK3/MAP3K11, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has at least about 70% inhibition by one of a group of kinases: Aurora A, Aurora B, Aurora C, ROS/ROS1, ARK5/NUAK1, TNK1, STK16, CHK2 MLK1/MAP3K9 and MLK3/MAP3K11.

In some embodiments, the irreversible BTK inhibitor has at least about 75% inhibition of a kinase selected from the group consisting of Aurora A, Aurora B, ROS/ROS1, ARK5/NUAK1, TNK1, STK16, and MLK1/MAP3K9, or a combination thereof . In some embodiments, the irreversible BTK inhibitor has at least about 75% inhibition of a panel of kinases consisting of: Aurora A, Aurora B, ROS/ROS1, ARK5/NUAK1, TNK1, STK16, and MLK1/MAP3K9.

In some embodiments, the irreversible BTK inhibitor has at least about 80% inhibition of a kinase selected from the group consisting of Aurora A, Aurora B, ROS/ROS1, ARK5/NUAK1, TNK1, and MLK1/MAP3K9, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has at least about 80% inhibition by one of a group of kinases: Aurora A, Aurora B, ROS/ROS1, ARK5/NUAK1, TNK1, and MLK1/MAP3K9.

In some embodiments, the irreversible BTK inhibitor has at least about 85% inhibition of a kinase selected from the group consisting of Aurora A, Aurora B, ROS/ROS1, ARK5/NUAK1, and MLK1/MAP3K9, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has at least about 85% inhibition by one of a group of kinases: Aurora A, Aurora B, ROS/ROS1, ARK5/NUAK1, and MLK1/MAP3K9.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition comparable to a reference kinase inhibitor for one or more kinases selected from the group consisting of: TNK1, STK16, ABL1, AXL, TYK2, CHK2, MLK1/ MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, SIK1, PKCb2, and CLK2, or a combination thereof. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor is directed against a panel of kinases having a percent inhibition comparable to a reference kinase inhibitor: TNK1, STK16, ABL1, AXL, TYK2, CHK2, MLK1/MAP3K9, MLK2/ MAP3K10, MLK3/MAP3K11, SIK1, PKCb2, and CLK2, or a combination thereof. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the percent inhibition of the irreversible BTK inhibitor against one or more kinases selected from the group consisting of at least the observed percent inhibition of the reference kinase inhibitor: TNK1, STK16, ABL1, AXL , TYK2, CHK2, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, SIK1, PKCb2, and CLK2, or a combination thereof. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least at least a kinase selected from the group consisting of: About 80%, at least about 85%, at least about 90%, or at least about 95% inhibition: TNK1, STK16, ABL1, AXL, TYK2, CHK2, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, SIK1, PKCb2, and CLK2, Or a combination thereof.

In some embodiments, the irreversible BTK inhibitor has at least about 50% inhibition of a kinase selected from the group consisting of: TNK1, STK16, ABL1, AXL, TYK2, CHK2, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, SIK1 , PKCb2 and CLK2, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has at least about 50% inhibition of a panel of kinases consisting of: TNK1, STK16, ABL1, AXL, TYK2, CHK2, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11 , SIK1, PKCb2 and CLK2.

In some embodiments, the irreversible BTK inhibitor has at least about 55% inhibition of a kinase selected from the group consisting of: TNK1, STK16, ABL1, CHK2, MLK1/MAP3K9, and MLK3/MAP3K11, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has at least about 55% inhibition by one of a group of kinases: TNK1, STK16, ABL1, CHK2, MLK1/MAP3K9, and MLK3/MAP3K11, or a combination thereof.

In some embodiments, the irreversible BTK inhibitor has at least about 60%, at least about 65%, or at least about 70% inhibition of a kinase selected from the group consisting of: TNK1, STK16, CHK2, MLK1/MAP3K9 and MLK3/MAP3K11, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has at least about 60%, at least about 65%, or at least about 70% inhibition of one of the group consisting of: CHK2, MLK1/MAP3K9, and MLK3/MAP3K11.

In some embodiments, the irreversible BTK inhibitor has at least about 75% inhibition of a kinase selected from the group consisting of: TNK1, STK16, and MLK1/MAP3K9, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has at least about 75% inhibition of a panel of kinases consisting of: TNK1, STK16, and MLK1/MAP3K9.

In some embodiments, an irreversible BTK inhibitor for use in the invention has a percentage of inhibition comparable to a reference kinase inhibitor for one or more kinases selected from the group consisting of: c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS, or a combination thereof. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor is directed against a panel of kinases having a percent inhibition comparable to a reference kinase inhibitor: c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS, or a combination thereof. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition against one or more kinases selected from the group consisting of: c-Kit, PDGFRa, RIPK2 HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS, or a combination thereof. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has no more than about 50%, no more than about 45%, no more than about 40%, no more than about 35%, no more than about 30%, no more than a kinase selected from the group consisting of: More than about 25%, no more than about 20%, no more than about 15%, no more than about 10%, or no more than about 5% inhibition: c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK, LCK, ZAK/ MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS, or a combination thereof.

In some embodiments, the irreversible BTK inhibitor has no more than about 50%, no more than about 45%, no more than about 40%, no more than about 35%, no more than about 30%, no more than a kinase selected from the group consisting of: More than about 25%, no more than about 20%, no more than about 15%, no more than about 10%, or no more than about 5% inhibition: RIPK2, HCK, LYN, CSK, LCK, LYN B, and FYN, or a combination thereof.

In some embodiments, the irreversible BTK inhibitor has no more than about 50%, no more than about 45%, no more than about 40%, no more than about 35%, no more than about 30%, no more than a kinase selected from the group consisting of: More than about 25%, no more than about 20%, no more than about 15%, no more than about 10%, or no more than about 5% inhibition: EPHA6, LYN B, FRK/PTK5, RIPK3, BRAF, ARAF, and SRMS, or combinations thereof .

In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of CLL/SLL comprising administering to a patient in need thereof an irreversible BTK inhibitor and lenalidomide, wherein the irreversible BTK inhibitor At least about 50% inhibition of a kinase selected from the group consisting of c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, BRAF, RIPK3, ARAF and SRMS, or a combination thereof.

In some embodiments, the irreversible BTK inhibitor has no more than about 50% inhibition of a kinase selected from the group consisting of c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has no more than about 50% inhibition of one of the group consisting of: c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS.

In some embodiments, the irreversible BTK inhibitor has no more than about 45% inhibition of a kinase selected from the group consisting of c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B , FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has no more than about 45% inhibition of one of the group consisting of: c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS.

In some embodiments, the irreversible BTK inhibitor has no more than about 40% inhibition of a kinase selected from the group consisting of c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B , FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has no more than about 40% inhibition by one of a group of kinases: c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS.

In some embodiments, the irreversible BTK inhibitor has no more than about 35% inhibition of a kinase selected from the group consisting of: c-Kit, RIPK2, HCK, EPHA6, LYN, CSK, ZAK/MLTK, LYN B, FRK/PTK5 , FYN, RIPK3, BRAF, ARAF and SRMS, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has the following No more than about 35% inhibition of one of the various components of the kinase: c-Kit, RIPK2, HCK, EPHA6, LYN, CSK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF, and SRMS.

In some embodiments, the irreversible BTK inhibitor has no more than about 30% inhibition of a kinase selected from the group consisting of c-Kit, RIPK2, HCK, EPHA6, LYN, CSK, ZAK/MLTK, LYN B, FRK/PTK5 , FYN, RIPK3, BRAF, ARAF and SRMS, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has no more than about 30% inhibition of one of the group consisting of: c-Kit, RIPK2, HCK, EPHA6, LYN, CSK, ZAK/MLTK, LYN B, FRK /PTK5, FYN, RIPK3, BRAF, ARAF and SRMS.

In some embodiments, the irreversible BTK inhibitor has no more than about 25% inhibition of a kinase selected from the group consisting of: c-Kit, RIPK2, EPHA6, CSK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3 , BRAF, ARAF and SRMS, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has no more than about 25% inhibition of one of the group consisting of: c-Kit, IPK2, EPHA6, CSK, ZAK/MLTK, LYN B, FRK/PTK5, FYN , RIPK3, BRAF, ARAF and SRMS.

In some embodiments, the irreversible BTK inhibitor has no more than about 20% inhibition of a kinase selected from the group consisting of: EPHA6, CSK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF, and SRMS , or a combination thereof. In some embodiments, the irreversible BTK inhibitor has no more than about 20% inhibition of one of the group consisting of: EPHA6, CSK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF And SRMS.

In some embodiments, the irreversible BTK inhibitor has no more than about 15% inhibition of a kinase selected from the group consisting of: EPHA6, LYN B, FRK/PTK5, RIPK3, BRAF, ARAF, and SRMS, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has No more than about 15% inhibition by one of the following groups of kinases: EPHA6, LYN B, FRK/PTK5, RIPK3, BRAF, ARAF, and SRMS.

In some embodiments, the irreversible BTK inhibitor has no more than about 10% inhibition of a kinase selected from the group consisting of: EPHA6, LYN B, FRK/PTK5, RIPK3, BRAF, ARAF, and SRMS, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has no more than about 10% inhibition by one of a group of kinases: EPHA6, LYN B, FRK/PTK5, RIPK3, BRAF, ARAF, and SRMS.

In some embodiments, the irreversible BTK inhibitor has no more than about 5% inhibition of a kinase selected from the group consisting of: EPHA6, FRK/PTK5, RIPK3, BRAF, ARAF, and SRMS, or a combination thereof. In some embodiments, the irreversible BTK inhibitor has no more than about 5% inhibition by one of the group consisting of: EPHA6, FRK/PTK5, RIPK3, BRAF, ARAF, and SRMS.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of LYN comparable to a reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of no more than the observed KIN of the reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of LYN of about 20-30%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of LYN of about 25-30%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of LYN of about 25-28%. In some embodiments, the irreversible BTK inhibitor has no more than about 25%, no more than about 26%, no more than about 27%, no more than about 28%, no more than about 29%, no more than about 30%, no more than about Percent inhibition of LYN of 31%, no more than about 32%, or no more than about 33%. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of c-Kit comparable to a reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of c-Kit that does not exceed the observed reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of c-Kit of about 15-25%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of c-Kit of about 20-25%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of c-Kit of about 20-23%. In some embodiments, the irreversible BTK inhibitor has no more than about 15%, no more than about 16%, no more than about 17%, no more than about 18%, no more than about 19%, no more than about 20%, no more than about Percent inhibition of c-Kit of 21%, no more than about 22%, no more than about 23%, no more than about 24%, or no more than about 25%. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of PDGFRa comparable to a reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of PDGFRa that does not exceed the observed reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of PDGFRa of about 30-40%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of PDGFRa of about 35-40%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of PDGFRa of between about 35 and 38%. In some embodiments, the irreversible BTK inhibitor has no more than about 30%, no more than about 31%, no more than about 32%, no more than about 33%, no more than about 34%, no more than about 35%, no more than about Percent inhibition of PDGFRa of 36%, no more than about 37%, no more than about 38%, no more than about 39%, or no more than about 40%. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of RIPK2 comparable to a reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of RIPK2 that does not exceed the observed reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of RIPK2 of about 20-30%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of RIPK2 of from about 20% to about 25%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of RIPK2 of about 22-25%. In some embodiments, the irreversible BTK inhibitor has no more than about 18%, no more than about 19%, no more than about 20%, no more than about 21%, no more than about 22%, no more than about 23%, no more than about Percent inhibition of RIPK2 of 24%, no more than about 25%, no more than about 26%, or no more than about 27%. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of HCK comparable to a reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of HCK that does not exceed the observed reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of HCK of between about 25 and 35%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of HCK of about 27-32%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of HCK of about 28-31%. In some embodiments, the irreversible BTK inhibitor has no more than about 26%, no more than about 27%, no more than about 28%, no more than about 29%, no more than about 30%, no more than about 31%, no more than about Percent inhibition of HCK of 32%, no more than about 33% or no more than about 34%. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of EPHA6 comparable to a reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of EPHA6 that does not exceed the observed reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of about 0-10% EPHA6. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of from about 0% to about 5% of EPHA6. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of from about 0% to about 3% of EPHA6. In some embodiments, the irreversible BTK inhibitor has no more than about 0.5%, no more than about 0.6%, no more than about 0.7%, no more than about 0.8%, no more than about 0.9%, no more than about 1%, no more than about Percent inhibition of 25%, no more than about 3%, or no more than about 4% of EPHA6. In some embodiments, the kinase inhibitor is the reference compound 2. In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of CSK comparable to a reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of CSK that does not exceed the observed reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of CSK of about 10-20%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of CSK of about 15-20%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of CSK of about 16-19%. In some embodiments, the irreversible BTK inhibitor has no more than about 15%, no more than about 16%, no more than about 17%, no more than about 18%, no more than about 19%, no more than about 20%, no more than about Percent inhibition of CSK of 21%, no more than about 22%, or no more than about 23%. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of LCK comparable to a reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of LCK that does not exceed the observed reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of LCK of about 30-40%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of LCK of about 32-37%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of LCK of about 34-37%. In some embodiments, the irreversible BTK inhibitor has no more than about 34%, no more than about 35%, no more than about 36%, no more than about 37%, no more than about 38%, no more than about 39%, no more than about Percent inhibition of LCK of 40%, no more than about 41% or no more than about 42%. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of ZAK/MLTK comparable to a reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of ZAK/MLTK that does not exceed the observed reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of ZAK/MLTK of about 10-20%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of ZAK/MLTK of about 12-17%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of ZAK/MLTK of about 14-17%. In some embodiments, the irreversible BTK inhibitor has no more than about 12%, no more than about 13%, no more than about 14%, no more than about 15%, no more than about 16%, no more than about 17%, no more than about Percent inhibition of ZAK/MLTK of 18%, no more than about 19% or no more than about 20%. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of LYN B comparable to a reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of LYN B that does not exceed the observed reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of LYN B of from about 0% to about 10%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of LYN B of from about 3 to 8%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of about 4-7% of LYN B. In some embodiments, the irreversible BTK inhibitor has no more than about 1%, no more than about 2%, no more than about 3%, no more than about 4%, no more than about 5%, no more than about 6%, no more than about Percent inhibition of 7% B of 7%, no more than about 8%, no more than about 9%, or no more than about 10%. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of FRK/PTK5 comparable to a reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of FRK/PTK5 that does not exceed the observed reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of FRK/PTK5 of from about 0% to about 10%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of FRK/PTK5 of from about 0% to about 5%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of FRK/PTK5 of from about 0% to about 3%. In some embodiments, the irreversible BTK inhibitor has no more than about 0.5%, no more than about 0.6%, no more than about 0.7%, no more than about 0.8%, no more than about 0.9%, no more than about 1%, no more than about Percent inhibition of FRK/PTK5 of 1.5%, no more than about 2%, no more than about 3%, or no more than about 4%. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of FYN comparable to a reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of no more than the observed FYN of the reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of FYN of about 15-25%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of FYN of about 15-20%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of FYN of about 17-20%. In some embodiments, the irreversible BTK inhibitor has no more than about 15%, no more than about 16%, no more than about 17%, no more than about 18%, no more than about 19%, no more than about 20%, no more than about Percent inhibition of FYN of 21%, no more than about 22%, or no more than about 23%. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition of BRAF comparable to a reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of BRAF that does not exceed the observed reference kinase inhibitor. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of BRAF of from about 0% to about 10%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of BRAF of from about 0.1% to about 5%. In some embodiments, the irreversible BTK inhibitor has a percent inhibition of BRAF of from about 0.2% to about 3%. In some embodiments, the irreversible BTK inhibitor has no more than about 0.1%, no more than about 0.2%, no more than about 0.3%, no more than about 0.4%, no more than about 0.5%, no more than about 0.6%, no more than about Percent inhibition of BRAF of 0.7%, no more than about 0.8%, no more than about 0.9%, no more than about 1%, no more than about 2%, no more than about 3%, no more than about 4%, or no more than about 5%. In some embodiments, the reference kinase inhibitor is Compound 2 . In some embodiments, the percent inhibition of the reference kinase inhibitor is the percent inhibition exhibited by Compound 2 in Example 3.

Compound 1 is an irreversible BTK inhibitor

As noted above, Bruton's tyrosine kinase (Btk) is a non-receptor tyrosine kinase that is primarily restricted to B lymphocytes, monocytes, and limiting cell expression of mast cells or basophils. Btk is a key component of the B cell receptor (BCR) signaling network and is critical for B cell development. Investigations have shown that some B cell malignancies (including CLL/SLL) are dependent on BCR signaling, suggesting that disrupting this signaling can be a promising therapeutic opportunity. Recently, agents that inhibit spleen tyrosine kinase (Syk) and Btk (ie, two components of the BCR signaling pathway) have been reported for use in various B-cell non-Hodgkin's lymphomas (NHL) and CLL/SLL. Clinical anti-tumor response.

Compound 1 is active in a variety of assays and therapeutic models, demonstrating covalent and irreversible inhibition of BTK (in enzymatic and cellular assays). Compound 1 by high affinity binding to the Btk adenosine triphosphate (ATP) binding site and forms a Btk target protein covalently bound to inhibition of Btk activity, in vitro and in vivo to provide rapid, complete and prolonged inhibition of Btk activity.

Inhibition of autophosphoric acid on Btk in Ramos cells (human Burkitt lymphoma cell line) by Compound 1 at an effective concentration of 1 nM to 10 nM required to achieve 50% inhibition (EC ₅₀ ) Phosphorylation of the Btk reaction site (Tyr1217) at the chemistry site (Tyr223) and PLCγ2. Compound 1 shows high selectivity for related kinases in cell analysis systems.

In a single-dose study of healthy individuals, Compound 1 was shown to be sufficiently safe to predict pharmacokinetics (PK) and at a dose greater than 0.5 mg/kg, Btk receptor target occupancy in normal human peripheral blood B cells was 80. % to 100%. A phase I dose escalation study of single agent compound 1 is currently underway for different hematological malignancies (including CLL/SLL).

Lenalidomide

Lenalidomide (REVLIMID®) belongs to the class of pharmaceutical compounds known as immunomodulatory drugs (IMiDsP®). The key to its therapeutic potential lies in the fact that it has multiple mechanisms of action that act to produce anti-inflammatory and anti-tumor effects. These effects depend on the cell type and trigger stimuli. To date, lenalidomide has been associated with tumor necrosis factor (TNF)-alpha inhibition, T cell co-stimulation, anti-proliferative and anti-angiogenic activities.

Lenalidomide has been approved by several global health authorities (including the United States Food and Drug Administration, but not the European Union) for the genetic abnormality of the 5q cell Treatment of patients with low- or medium-risk-1 myelodysplastic syndromes with or without transfusion-dependent anemia associated with other cytogenetic abnormalities. Lenalidomide has also been approved by several global health authorities (notably US and EU) for use in combination with dexamethasone for patients with previously treated multiple myeloma (MM).

Lenalidomide is being investigated as a therapeutic agent for a variety of oncological indications including MM, NHL and solid tumors. Non-oncological indications have also been studied.

Tumor necrosis factor-α has been proposed as an autocrine growth factor that crosses the TNF-α receptor on CLL cells. High serum vascular endothelial growth factor (VEGF) levels and CLL cells have also been reported Vascular Endothelial Growth Factor Receptor 1 (VEGFR-1) and Vascular Endothelial Growth Factor Receptor 2 (VEGFR-2). Lenalidomide has been shown to have the ability to regulate and regulate growth factors/cytokines including VEGF, TNF-[alpha], interleukin (IL)-10 and IL-6. Immunomodulatory drugs have been shown to synergistically stimulate immune effector cells (T cells and natural killer cells). These observations and the known activity of thalidomide and lenalidomide in another B-cell malignant multiple myeloma provide an initial rationale for studying lenalidomide in CLL/SLL.

The single agent lenalidomide has been studied in relapsed/refractory CLL. Individuals were given lenalidomide 25 mg for 21 days per 28 day cycle. The overall response rate was 53% (18% complete response [CR], 36% partial reaction [PR]). Clinical response (CR or was observed in 47% of individuals with del(11)(q23) and/or del(17)(p13.1) unfavorable cytogenes and 30% of fludarabine refractory individuals PR). The median time to optimal response was 5.9 months (1.6-18.3 months) and the median progression-free survival (PFS) time was 19.4 months (1.2 to 31.8 months). In addition, individuals with relapsed/refractory CLL were treated by daily administration of lenalidomide over a 28 day period until disease progression.

A large number of clinical data indicate that lenalidomide is active in relapsed/refractory aggressive B cell malignancies. In addition, the B cell receptor (BCR) pathway of Btk as a downstream component has been linked to the pathogenesis of B cell proliferation, activation and survival. The role of Btk in CLL/SLL is supported by early reports of clinical studies involving Btk inhibitors.

anti-CD20 antibody

CD20 (the first B cell-specific antigen defined by the monoclonal antibody tositumomab) plays a key role in B cell development. Human CD20 is a phosphoprotein having 297 amino acids (30 to 35 kDa) in four transmembrane domains encoded by the gene MS4A1 located on chromosome 11q12.2. CD20 plays a key role in B cell development and is a biomarker for immunotherapy targeting B cell-derived diseases. CD20 is an integral membrane protein expressed by early differentiation of B lymphocytes and by most B cell lymphomas, but not by differentiated plasma cells. CD20 remains on the B cell membrane after antibody binding without dissociation or internalization. Despite the binding to the Src family of tyrosine kinases (such as Lyn, Fyn, and Lck), CD20 still functions, and thus is involved in the phosphorylation cascade of intracellular proteins. Anti-CD20 antibodies are broadly classified into type I and type II antibodies. Both types of anti-CD20 antibodies show considerable ability to activate Fc-FcyR interactions, such as antibody-dependent cellular cytotoxicity (ADCC) and phagocytosis. Type I anti-CD20 antibodies redistribute CD20 in membrane lipid rafts and potently activate complement dependent cytotoxicity (CDC). Type II anti-CD20 antibodies weakly activate CDC, but more potently induce direct progressive cell death.

In some embodiments, the invention contemplates that a BTK inhibitor (eg, Compound 1 ), together with lenalidomide and an anti-CD20 antibody, is useful for treating a BTK-mediated disease or disorder. Accordingly, in some embodiments, the invention comprises a method of treating a BTK mediated disease or disorder, the method comprising administering to each patient in need thereof a compound 1 , a lenalidomide, and an anti-CD20 antibody. Those skilled in the art will readily be able to identify and select additional anti-CD20 antibodies suitable for use in the present invention. For example, in some embodiments, such antibodies are described in, for example, U.S. Patent Nos. 8,153,125, 8,147,832, 8,101,179, 8,084,582, 8,057,793, and 7,879,984, and U.S. Patent Publication No. 2011/ No. 0129412, No. 2012/0183545, No. 2012/0134990 and No. 2012/0034185.

In some embodiments, the anti-CD20 antibody used in the invention is a type I antibody. In some embodiments, the anti-CD20 antibody used in the invention is a Type II antibody.

In some embodiments, the anti-CD20 antibody is an antibody that binds to a CD20 epitope selected from the group consisting of ¹⁷⁰ ANPS ¹⁷³ and ¹⁸² YCYSI ¹⁸⁵ .

In some embodiments, the anti-CD20 antibody binding affinity of the epitope of the CD20 antigen (K _d) of less than 12nM, less than 11 nM, less than 10 nM, less than 9nM, less than 8nM, less than 7 nM, less than 6nM, less than of 5 nM, less than 4nM, less than 3nM, less than 2nM or less than 1nM.

Rituximab is only one example of an anti-CD20 antibody. In some embodiments, anti-CD20 antibodies for use in the invention include, for example, rituximab (Rituxan® or MabThera®), Gazyva® (i.e., obinutuzumab), and Arzerra® (Orpheus Anti (ofatumumab)). For ease of reference, the methods provided and the protocols detailed herein refer to exemplary anti-CD20 antibodies (i.e., rituximab); however, this reference is not intended to limit the invention to a single anti-CD20 antibody. In fact, those of skill in the art will read all references to rituximab or a biological analog thereof to encompass classes of anti-CD20 antibodies. For example, it should be understood that in each case where rituximab is mentioned, the anti-CD20 antibody arfazumab (Arzerra®) or oubizumab (Gazyva®) may be administered instead. Accordingly, in some embodiments, the methods provided comprise administering Compound 1 , lenalidomide, and/or orfarizumab. In some of these embodiments, orfarizumab was administered in 12 doses according to the following schedule: 300 mg first dose, followed by 7 doses of 2000 mg dose per week after 1 week, followed by 4 weeks, 4 doses of 2000 mg dose every 4 weeks . In some embodiments, the methods provided comprise administering Compound 1 , lenalidomide, and/or opiconumab. In some of these examples, the following two 28-day cycles were administered to opiconumab: 100 mg on the first day of the first cycle; 900 mg on the second day of the first cycle; on the eighth day of the first cycle and 1000 mg for 15 days; and 1000 mg on the first day of the 2-6th cycle. Thus, in some embodiments, the term "rituximab" encompasses the requirement to obtain a sales authorization for the same or biosimilar product in a country or territory selected from a group of countries consisting of the United States, Europe, and Japan. All corresponding anti-CD20 antibodies.

In some embodiments, the anti-CD20 antibody has the same or similar activity as rituximab or a biological analog thereof. In some embodiments, the anti-CD20 antibody binds to a region or epitope that is identical or similar to rituximab or a fragment thereof. In some embodiments, the anti-CD20 antibody competes with rituximab or a fragment thereof for binding to CD20. In some embodiments, the anti-CD20 antibody is bioequivalent to rituximab or a fragment thereof. In some embodiments, the anti-CD20 antibody is a biological analog of rituximab or a fragment thereof. In some implementations In one embodiment, the anti-CD20 antibody is a variant or derivative of rituximab, including a functional fragment, derivative or antibody conjugate.

Rituximab

Rituxan® or MabThera® is a genetically engineered CD20 cell surface molecule present in normal B lymphocytes and B cell CLL and in most forms of non-Hodgkin's B cell lymphoma. Cell lytic chimeric murine/human monoclonal IgG ₁ κ antibody. Rituximab has a binding affinity of about 8.0 nM for the CD20 antigen. Rituximab induces complement-dependent cytotoxicity (CDC) and antibody-dependent cellular cytotoxicity (ADCC), resulting in its clinical activity against lymphoma cells. Rituximab also causes B cell apoptosis upon binding to CD20, thereby directly inhibiting cell growth.

Rituximab is produced from a suspension culture of mammalian cells (Chinese hamster ovaries) in a nutrient medium containing the antibiotic gentamicin. Gentamicin is not detectable in the final product. Rituximab is a sterile, clear, colorless, preservative-free liquid concentrate for intravenous administration. Rituximab was supplied at a concentration of 10 mg/mL in a single use vial of 100 mg/10 mL or 500 mg/50 mL. Rituximab was formulated in polysorbate 80 (0.7 mg/mL), sodium citrate dihydrate (7.35 mg/mL), sodium chloride (9 mg/mL), and water for injection. Rituxan® (or MabThera®) has a pH of 6.5.

Rituximab has been investigated in clinical studies and approved for the treatment of patients with CLL with fludarabine and cyclophosphamide, as well as with methotrexate A patient with rheumatoid arthritis. Rituximab is also approved for the treatment of non-Hodgkin's lymphoma, Wegener's Granulomatosis and Microscopic Polyangiitis. In addition, studies have shown that rituximab and bendamustine are also effective in the treatment of relapsed or refractory CLL and NHL, such as indolent lymphoma and mantle cell lymphoma. It is understood and understood that when rituximab is administered with Compound 1 and lenalidomide, rituximab can be used as a single agent or with (i) fludarabine and cyclophosphamide ("FCR"") or (ii) Bendamustine ("BR") is used together. Accordingly, in some embodiments, the methods provided comprise administering each of Compound 1 , lenalidomide, rituximab, fludarabine, and cyclophosphamide. In some embodiments, the methods provided comprise administering each of Compound 1 , lenalidomide, rituximab, and bendamustine.

I. General dosing plan

As described herein, the methods provided comprise administering two or more therapeutic agents (i.e., Compound 1 , lenalidomide, and/or anti-CD20 antibodies). It should be understood, each therapeutic agent can be synchronized as part of the program or sequence of administration (e.g. Compound 1 may lenalidomide and / or anti-CD20 antibody before, during or after the administration, and vice-versa) and administered. For example, Compound 1 can be administered one or more hours, one or more days, or one or more weeks prior to administration of lenalidomide or an anti-CD20 antibody. In other embodiments, each of Compound 1 and lenalidomide can be administered one or more hours, one or more days, or one or more weeks prior to administration of the anti-CD20 antibody. In other embodiments, each of Compound 1 and the anti-CD20 antibody can be administered one or more hours, one or more days, or one or more weeks prior to administration of lenalidomide.

In some embodiments, the invention provides methods for treating, stabilizing or ameliorating the severity or progression of one or more diseases or conditions associated with BTK. In some embodiments, the invention provides methods for preventing progression of a disease or disorder associated with BTK. In some embodiments, the disease or disorder associated with BTK is selected from the group consisting of chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL).

In some embodiments, the disease or disorder associated with BTK is refractory CLL. In some embodiments, the disease or disorder associated with BTK is recurrent CLL. In some embodiments, the disease or disorder associated with BTK is a refractory SLL. In some embodiments, the disease or disorder associated with BTK is a recurrent SLL.

In some embodiments, the methods provided comprise administering a combination of Compound 1 and lenalidomide to a patient in need thereof. In some such embodiments, Compound 1 and lenalidomide in each composition additionally comprises one or more pharmaceutically acceptable excipients and administered in the form of. In some embodiments, the methods provided additionally comprise administering an anti-CD20 antibody, such as rituximab. In some embodiments, the methods provided comprise administering each of Compound 1 , lenalidomide, and an anti-CD20 antibody (eg, rituximab).

In some embodiments, a method is provided comprising administering to a patient in need thereof a therapeutically effective amount of Compound 1 in combination with a therapeutically effective amount of lenalidomide. Accordingly, in some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of one or more diseases associated with BTK, the method comprising administering to a patient in need thereof a therapeutically effective amount of Compound 1 and A combination of a therapeutically effective amount of lenalidomide. In some embodiments, the methods provided additionally comprise administering an anti-CD20 antibody, such as rituximab. In some embodiments, the methods provided comprise administering a therapeutically effective amount of each of Compound 1 , lenalidomide, and an anti-CD20 antibody (eg, rituximab).

In some embodiments, the methods provided comprise administering a combination of Compound 1 and lenalidomide, wherein Compound 1 and lenalidomide are administered once daily ("QD"). In some embodiments, the methods provided comprise administering a combination of Compound 1 and lenalidomide, wherein Compound 1 is administered twice daily ("BID"). In some embodiments, the methods provided additionally comprise administering an anti-CD20 antibody, such as rituximab. In some embodiments, the methods provided comprise administering each of Compound 1 , lenalidomide, and an anti-CD20 antibody (eg, rituximab), wherein Compound 1 is administered BID. In some of these embodiments, lenalidomide is administered to QD. For the sake of clarity, administration of a 375 mg dose of Compound 1 "BID" means that two separate doses of 375 mg are administered to the patient within one day.

In some embodiments, the methods provided comprise administering a combination of Compound 1 and lenalidomide, wherein lenalidomide is administered once daily. In some embodiments, the methods provided comprise administering a combination of Compound 1 and lenalidomide, wherein Compound 1 is administered twice daily and lenalidomide is administered once daily. In some embodiments, the methods provided additionally comprise administering an anti-CD20 antibody, such as rituximab, wherein the anti-CD20 antibody is administered once in a 28 day period. In some of these embodiments, the anti-CD20 antibody is administered on the first or second day of the first cycle. In some of these examples, the anti-CD20 antibody was administered on day 1 of the 28 day cycle. In some embodiments, the anti-CD20 antibody is administered on day 1 of cycles 2-6. In some embodiments, the methods provided comprise administering each of Compound 1 , lenalidomide, and an anti-CD20 antibody, wherein the anti-CD20 antibody is administered once in a 28 day period. In some of these embodiments, the anti-CD20 antibody is administered on day 1 or day 2 of the 28 day cycle. It should be appreciated that while the method herein refers to administration of the compound 1, but these methods are equally applicable to the administration of the compound in the form of a salt (e.g. benzenesulfonate salt of Compound 1) method.

In some embodiments, Compound 1 and lenalidomide are each administered as a pharmaceutically acceptable composition. In some such embodiments, each pharmaceutically acceptable composition is formulated into an oral dosage form. In some embodiments, the oral dosage forms are capsules. In some embodiments, the methods provided additionally comprise a pharmaceutically acceptable composition for administration of an anti-CD20 antibody (eg, rituximab). In some embodiments, a method provided comprises administering a pharmaceutically acceptable composition of each of Compound 1 , lenalidomide, and an anti-CD20 antibody (eg, rituximab). In some such embodiments, a pharmaceutically acceptable composition of an anti-CD20 antibody is formulated into an intravenous composition.

In some embodiments, the pharmaceutically acceptable composition comprising Compound 1 comprises from about 5% to about 60% of Compound 1 or a pharmaceutically acceptable salt thereof, based on the total weight of the composition. In some embodiments, the pharmaceutically acceptable composition comprising Compound 1 comprises from about 5% to about 15%, or from about 7% to about 15%, or from about 7% to about, based on the total weight of the composition. 10%, or about 9% to about 12% of Compound 1 . In some embodiments, the methods provided comprise administering to a patient in need thereof a pharmaceutically acceptable composition comprising from about 25% to about 75%, or about 30, based on the total weight of the formulation. From about 60%, or from about 40% to about 50%, or from about 40% to about 45% of Compound 1 . In certain embodiments, the regimen provided comprises administering to a patient in need thereof a pharmaceutically acceptable composition comprising about 6%, based on the total weight of the given composition or formulation. 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 20%, about 30%, about 40%, about 41%, about 42%, about 43% About 44%, about 45%, about 50%, about 60%, about 70%, or about 75% of Compound 1 .

In some embodiments, the pharmaceutically acceptable composition comprising lenalidomide comprises from about 2.5% to about 6.25% lenalidomide, based on the total weight of the composition. In some embodiments, the pharmaceutically acceptable composition comprises about 2.5%, about 3.75%, or about 6.25% lenalidomide or a pharmaceutically acceptable salt thereof, based on the total weight of the composition. In some embodiments, the composition of lenalidomide is a composition commercially available from Celgene Corporation. In some embodiments, the pharmaceutically acceptable composition of lenalidomide is the composition described in Table 2 or Table 3.

Rituximab is commercially available as a 10 mg/mL solution comprising sodium citrate, polysorbate 80, sodium chloride, sodium hydroxide, hydrochloric acid and water. Commercially available vials contain 100 mg/10 mL or 500 mg/50 mL.

In some embodiments, the pharmaceutically acceptable composition comprises from about 1 mg/mL to about 4 mg/mL rituximab. In some embodiments, the pharmaceutically acceptable composition comprises about 1 mg/mL, about 2 mg/mL, about 3 mg/mL, or about 4 mg/mL rituximab. In some embodiments, the pharmaceutically acceptable composition comprises 10 mg/mL.

In some embodiments, the methods provided comprise administering a combination of Compound 1 and lenalidomide daily for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27 or 28 days. In some embodiments, the treatment regimen comprises at least one 28 day period. As used herein, the term "28 day cycle" is intended to mean that a patient in need thereof is administered a combination of Compound 1 and lenalidomide for 28 consecutive days. In some embodiments, the combination of Compound 1 and lenalidomide is administered once a day for at least one 28 day period. In some embodiments, the combination of Compound 1 and lenalidomide is administered for at least two, at least three, at least four, at least five, or at least six 28 day cycles. In some embodiments, the combination of Compound 1 and lenalidomide is administered once a day for at least seven, at least eight, at least nine, at least ten, at least eleven, or at least twelve 28-day cycles. . In some embodiments, the combination of Compound 1 and lenalidomide is administered once a day for at least thirteen, at least fourteen, at least fifteen, at least sixteen, at least seventeen, at least eighteen At least nineteen or at least twenty 28-day cycles. In some embodiments, a combination of Compound 1 and lenalidomide is administered to a patient for a lifetime. In some embodiments, Compound 1 is administered on Day 1 to Day 28 of one or more 28-day cycles (eg, once daily or twice daily) and is 1 in one or more 28-day cycles Lenalidomide is administered on day 21 (eg, once daily). In some embodiments, Compound 1 is administered from day 8 to day 28 of one or more 28 day cycles, and lenalidomide is administered from day 1 to day 28 of one or more 28 day cycles. . In some embodiments, Compound 1 is administered from Day 1 to Day 28 of one or more 28-day cycles, and lenalidomide is administered on Days 1 to 28 of one or more 28-day cycles. .

In some embodiments, each of Compound 1 and lenalidomide is administered to the patient for one, two or three 28 day cycles, followed by a reduction in the dose of lenalidomide. In some embodiments, each of Compound 1 and lenalidomide is administered to the patient for one, two or three 28 day cycles, followed by a reduction in the dose of lenalidomide to the previous group dose. In some embodiments, each of Compound 1 and lenalidomide is administered to the patient until the desired response (eg, complete response) is reached, followed by a reduction in the dose of lenalidomide.

In some embodiments, each of Compound 1 , lenalidomide, and rituximab is administered for at least six 28-day cycles, and each of Compound 1 and lenalidomide is additionally administered. Continue for at least one 28-day cycle. In some embodiments, each of Compound 1 , lenalidomide, and rituximab is administered for at least six 28-day cycles, and each of Compound 1 and lenalidomide is additionally administered. Then, continue for two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen or fourteen 28-day cycles. In some embodiments, each of Compound 1 , lenalidomide, and rituximab is administered for at least six 28-day cycles, and each of Compound 1 and lenalidomide is additionally administered. For the rest of the patient's life. In some embodiments, each of Compound 1 and lenalidomide is administered from Day 1 to Day 28 of one or more 28-day cycles (eg, once daily or twice daily), and Rituximab was administered on the first day of the 28-day cycle. In some embodiments, each of Compound 1 and lenalidomide is administered from Day 1 to Day 28 of one or more 28-day cycles, and on Day 1 or Day 2 of the 28-day cycle Invested in rituximab.

In some embodiments, each of Compound 1 and rituximab is administered to a patient in one or more cycles, followed by administration of Compound 1 , Rituximab, and Lenazide to the patient over a subsequent cycle. Each of the amines. In some embodiments, each of Compound 1 and rituximab is administered to the patient during the first cycle, followed by administration of Compound 1 , Rituximab, and Laina to the patient during Cycles 2-6. Each of the amines. In some embodiments, the methods provided comprise (i) administering to each of the patients in need thereof, Compound 1 and rituximab, during the first 28-day period; (ii) at 2-6 Each of Compound 1 , Rituximab, and Lenalidomide is administered to the patient during the cycle; and (iii) Compound 1 is administered to the patient over a period of one or more 28 days thereafter and/or Amine.

In some embodiments, two adjacent 28 day periods may be separated by a rest period. Such rest periods may be one day, two days, three days, four days, five days, six days, seven days or more, during which no one of Compound 1 and lenalidomide is administered to the patient or Both. In a preferred embodiment, two adjacent 28 day periods are continuous.

In some embodiments, a method is provided comprising administering to a patient in need thereof a combination of Compound 1 and lenalidomide, wherein the patient has failed at least one prior therapy. In some embodiments, a method is provided comprising administering to a patient in need thereof each of Compound 1 , lenalidomide, and rituximab, wherein the patient has failed at least one prior therapy.

Unit dosage form

The pharmaceutical compositions used in the present invention can be prepared in unit dosage form. One of ordinary skill in the art will appreciate that the unit dosage form described herein refers to the amount of the component in its free base form. It will be further understood by those skilled in the art that when the pharmaceutical composition comprises a salt form of one component (e.g., the besylate form of Compound 1 ), the amount of the salt form present in the composition is the same as the component ( That is, the unit dose equivalent amount of the free base of Compound 1 ). For example, a pharmaceutical composition comprising a besylate salt of Compound 1 will contain the 34.97 mg besylate salt form necessary to deliver an equivalent equivalent of 25 mg unit dose of Compound 1 free base.

In some embodiments, the method comprises providing to a patient in need there of administering a therapeutically effective amount of a compound, wherein the therapeutically effective amount of a compound is from about 250mg to about 1250mg. In some embodiments, a therapeutically effective amount of Compound 1 is administered in one or more discrete dosage forms. For example, in some embodiments, the therapeutically effective amount of Compound 1 is 250 mg, wherein the therapeutically effective amount is administered twice daily (BID) in the form of 125 mg. In some embodiments, the therapeutically effective amount of Compound 1 is 500 mg, wherein the therapeutically effective amount is administered twice daily (BID) in the form of 250 mg. In some embodiments, the therapeutically effective amount of Compound 1 is 750 mg, wherein the therapeutically effective amount is administered twice daily (BID) in the form of 375 mg. In some embodiments, the therapeutically effective amount of Compound 1 is 1000 mg, wherein the therapeutically effective amount is administered twice daily (BID) in the form of 500 mg.

In some embodiments, the methods provided comprise to have administered to treat a patient in need of an effective amount of a compound, wherein the compound therapeutically effective amount of a is from about 125mg to about 1250 mg, or from about 125mg to about 1125 mg, or about 125mg To about 1000 mg, or about 125 mg to about 875 mg, or about 125 mg to about 750 mg, or about 125 mg to about 625 mg, or about 125 mg to about 500 mg, or about 125 mg to about 375 mg, or about 125 mg to about 250 mg, or about 250 mg to About 1250 mg, or about 250 mg to about 1125 mg, or about 250 mg to about 1000 mg, or about 250 mg to about 875 mg, or about 250 mg to about 750 mg, or about 250 mg to about 625 mg, or about 250 mg to about 500 mg, or about 250 mg to about 375 mg, or from about 375 mg to about 1250 mg, or from about 375 mg to about 1125 mg, or from about 375 mg to about 1000 mg, or from about 375 mg to about 875 mg, or from about 375 mg to about 750 mg, or from about 375 mg to about 625 mg, or from about 375 mg to about 500 mg. Or from about 500 mg to about 1250 mg, or from about 500 mg to about 1125 mg, or from about 500 mg to about 1000 mg, or from about 500 mg to about 875 mg, or from about 500 mg to about 750 mg, or from about 500 mg to about 625 mg, or from about 625 mg to about 1250 mg Or from about 625 mg to about 1125 mg, or from about 625 mg to about 1000 mg, or about 625 mg to 875 mg, or from about 625 mg to about 750 mg, or from about 750 mg to about 1250 mg, or from about 750 mg to about 1125 mg, or from about 750 mg to about 1000 mg, or from about 875 mg to about 1250 mg, or from about 875 mg to about 1125 mg, or from about 875 mg to about 1000 mg .

In some embodiments, the methods provided comprise to have administered to treat a patient in need of an effective amount of a compound, wherein the compound of the therapeutically effective amount is about 125mg, 130mg, 135mg, 140mg, 145mg, 150mg, 155mg, 160mg 165mg, 170mg, 175mg, 180mg, 185mg, 190mg, 195mg, 200mg, 205mg, 210mg, 215mg, 220mg, 225mg, 230mg, 235mg, 240mg, 245mg, 250mg, 255mg, 260mg, 265mg, 270mg, 275mg, 280mg, 285mg 290mg, 295mg, 300mg, 305mg, 310mg, 315mg, 320mg, 325mg, 330mg, 335mg, 340mg, 345mg, 350mg, 355mg, 360mg, 365mg, 370mg, 375mg, 380mg, 385mg, 390mg, 395mg, 400mg, 405mg, 410mg 415mg, 420mg, 425mg, 430mg, 435mg, 440mg, 445mg, 450mg, 455mg, 460mg, 465mg, 470mg, 475mg, 480mg, 485mg, 490mg, 495mg, 500mg, 505mg, 510mg, 515mg, 520mg, 525mg, 530mg, 535mg 540mg, 545mg, 550mg, 555mg, 560mg, 565mg, 570mg, 575mg, 580mg, 585mg, 590mg, 595mg, 600mg, 605mg, 610mg, 615mg, 620mg, 625mg, 630mg, 635mg, 640mg, 645mg, 650mg, 65 5mg, 660mg, 665mg, 670mg, 675mg, 680mg, 685mg, 690mg, 695mg, 700mg, 705mg, 710mg, 715mg, 720mg, 725mg, 730mg, 735mg, 740mg, 745mg, 750mg, 755mg, 760mg, 765mg, 770mg, 775mg , 780mg, 785mg, 790mg, 795mg, 800mg, 805mg, 810mg, 815mg, 820mg, 825mg, 830mg, 835mg, 840mg, 845mg, 850mg, 855mg, 860mg, 865mg, 870mg, 875mg, 880mg, 885mg, 890mg, 895mg, 900mg 905mg, 910mg, 915mg, 920mg, 925mg, 930mg, 935mg, 940mg, 945mg, 950mg, 955mg, 960mg, 965mg, 970mg, 975mg, 980mg, 985mg, 990mg, 995mg, 1000mg, 1005mg, 1010mg, 1015mg, 1020mg, 1025mg 1030 mg, 1035 mg, 1040 mg, 1045 mg, 1050 mg, 1055 mg, 1060 mg, 1065 mg, 1070 mg, 1075 mg, 1080 mg, 1085 mg, 1090 mg, 1095 mg, 1100 mg, 1105 mg, 1110 mg, 1115 mg, 1120 mg, 1125 mg, 1130 mg, 1135 mg, 1140 mg, 1145 mg, 1150 mg 1,155 mg, 1160 mg, 1165 mg, 1170 mg, 1175 mg, 1180 mg, 1185 mg, 1190 mg, 1195 mg, 1200 mg, 1205 mg, 1210 mg, 1215 mg, 1220 mg, 1225 mg, 1230 mg, 1235 mg, 1240 mg, 1245 mg 1250mg.

In some embodiments, a method is provided comprising administering to a patient in need thereof a therapeutically effective amount of lenalidomide, wherein the therapeutically effective amount of lenalidomide is from about 2.5 mg to about 25 mg.

In some embodiments, the methods provided comprise administering to a patient in need thereof a therapeutically effective amount of lenalidomide, wherein the therapeutically effective amount of lenalidomide is from about 2.5 mg to about 25 mg, or about 2.5 mg to About 20 mg, or about 2.5 mg to about 15 mg, or about 2.5 mg to about 10 mg, or about 2.5 mg to about 7.5 mg, or about 2.5 mg to about 5 mg, or about 5 mg to about 25 mg, or about 5 mg to about 20 mg, Or from about 5 mg to about 15 mg, or from about 5 mg to about 10 mg, or from about 5 mg to about 7.5 mg, or from about 7.5 mg to about 25 mg, or from about 7.5 mg to about 20 mg, or from about 7.5 mg to about 15 mg, or about 7.5 mg To about 10 mg, or about 10 mg to About 25 mg, or from about 10 mg to about 20 mg, or from about 10 mg to about 15 mg, or from about 15 mg to about 25 mg, or from about 15 mg to about 20 mg, or from about 20 mg to about 25 mg.

In some embodiments, the methods provided comprise administering about 2.5 mg, 5 mg, about 7.5 mg, 10 mg, 15 mg, 20 mg, 25 mg of lenalidomide to a patient in need thereof.

In some embodiments, a method is provided comprising administering to a patient in need thereof a pharmaceutical composition comprising a unit dose of Compound 1 in combination with a unit dose of lenalidomide. In some such embodiments, the unit dosage of Compound 1 is 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, or about 250 mg.

In some embodiments, the unit dose of lenalidomide is about 2.5 mg, about 5 mg, about 7.5 mg, about 10 mg, about 15 mg, or about 25 mg.

In some embodiments, a method is provided comprising administering to a patient in need thereof a pharmaceutical composition comprising rituximab, wherein rituximab is administered as an infusion at a rate of 50 mg/h. In some embodiments, the infusion rate of rituximab is increased by 50 mg/h every 30 minutes to a maximum of 400 mg/h. In some embodiments, the infusion rate of rituximab is increased by 100 mg/h every 30 minutes to a maximum of 400 mg/h. Thus, in some embodiments, the infusion rate of rituximab is 100 mg/h. In some embodiments, the infusion rate of rituximab is 150 mg/h. In some embodiments, the infusion rate of rituximab is 200 mg/h. In some embodiments, the infusion rate of rituximab is 250 mg/h. In some embodiments, the infusion rate of rituximab is 300 mg/h. In some embodiments, the infusion rate of rituximab is 350 mg/h. In some embodiments, the infusion rate of rituximab is 400 mg/h.

II. Use of Compounds and Pharmaceutically Acceptable Compositions

The compounds 1 and compositions described herein are generally suitable for inhibiting the protein kinase activity of one or more enzymes. Examples of kinases that are inhibited by the compounds 1 and compositions described herein and which are suitable for use in the methods described herein include BTK and other TEC-kinases, including ITK, TEC, BMX, and RLK, or mutants thereof.

Bruton tyrosine kinase ("BTK", a member of TEC kinase) is a key signaling enzyme that is expressed in B lymphocytes, monocytes, and mast cells or basophils. BTK plays an essential role in the B cell signaling pathway that links cell surface B cell receptor (BCR) stimulation with downstream intracellular responses.

BTK is a key regulator of B cell development, activation, signaling and survival (Kurosaki, Curr. Op. Imm., 2000, 276-281; Schaeffer and Schwartzberg, Curr. Op . Imm . 2000, 282-288). In addition, BTK plays a role in many other hematopoietic signaling pathways, such as Toll like receptor (TLR) in macrophages and cytokine receptor-mediated TNF-α production, IgE in mast cells. Receptor (Fc_ε_RI) signaling, inhibition of Fas/APO-1 cell apoptosis signaling in B lineage lymphoid cells, and collagen-stimulated platelet aggregation. See, for example, CA Jeffries, et al, (2003), Journal of Biological Chemistry 278: 26258-26264; NJ Horwood, et al, (2003), The Journal of Experimental Medicine 197: 1603-1611; Iwaki et al. (2005), Journal of Biological Chemistry 280(48): 40261-40270; Vassilev et al. (1999), Journal of Biological Chemistry 274(3): 1646-1656 and Quek et al. (1998), Current Biology 8(20): 1137-1140.

Patients with BTK genetically inactivating mutations have significant barriers to B cell development, resulting in the almost complete absence of mature B lymphocytes and plasma cells, severely reducing Ig content and greatly inhibiting humoral responses to recall antigens (in Vihinen et al. Frontiers in Bioscience 5: reviewed in d917-928). Mice lacking BTK also had a reduced number of peripheral B cells and a greatly reduced serum IgM and IgG3 levels. Deletion of BTK in mice has a profound effect on anti-IgM-induced B cell proliferation and inhibits immune responses to non-thymus-dependent type II antigens (Ellmeier et al, J Exp Med 192:1611-1623 (2000)). BTK also plays a key role in the activation of mast cells via the high affinity IgE receptor (Fc_ε_RI). BTK-deficient murine mast cells reduce degranulation and reduce pro-inflammatory cytokine production following Fc_ε_RI cross-linking (Kawakami et al . Journal of Leukocyte Biology 65: 286-290).

Compound 1 is a BTK inhibitor and is therefore suitable for the treatment of one or more disorders associated with BTK activity. Accordingly, in some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of a BTK mediated disorder comprising the step of administering a combination of Compound 1 and lenalidomide to a patient in need thereof.

Chronic lymphocytic leukemia and small lymphocytic lymphoma

B cell dysregulation Chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL) indicate both ends of the same disease process range where blood/bone marrow involvement (CLL) and lymph node involvement (SLL) differ. CLL is a lymphoid proliferative malignancy characterized by progressive accumulation of morphologically mature but functionally incomplete lymphocytes in blood, bone marrow, and lymphoid tissues. It mainly affects older individuals with a median age of 65 to 70 years. The clinical course of CLL is between invasive disease with a long-term survival of more than 12 years and a median survival of 2 years.

Chronic lymphocytic leukemia is the most common leukemia in the United States and is usually characterized by immunophenotype as CD5+, CD23+, CD10-, CD19+, CD20 weak manifestation (dim), sIg weak manifestation, and cyclin D1- (the latter indicated with the set) The distinguishing feature of cell lymphoma). Chronic lymphocytic leukemia must also be distinguished from single B lymphocytosis (absolute single B cell count <5000/μL and no other clinical features of adenosis or lymphocytosis disorders). Recently, Lanasa, Furman, and the National Comprehensive Cancer Network NHL panel have reviewed the biology and prognostic factors of CLL/SLL and the advances in the development of a risk stratification approach to treat CLL/SLL.

The cell expression of Btk is limited and is mainly limited to B lymphocytes, monocytes and mast cells or basophils. Investigations have shown that some B-cell lymphomas and CLL/SLL are dependent on BCR signaling, suggesting that disrupting this signaling may be a promising therapeutic opportunity. near In addition, one of the CLLs has been reported to have in vitro BCR signaling, and immunoglobulin heavy chain gene somatic mutation (IgVH) is an important determinant of BCR response. In fact, the mutation status of BCR in CLL is one of the most powerful predictors of disease progression, as invasive disease usually shows a BCR encoded by an unmutated immunoglobulin variable heavy chain.

Mutant and unmutated CLL cells have been reported to be differentially responsive to ICR junctions in BCR, in which unmutated (rather than mutated) CLL cells respond by increasing overall tyrosine phosphorylation and up-regulating several genes involved in cell cycle regulation. Stimulated by BCR and allowed for cell growth and expansion. These data highlight the differential role of BCR signaling in the physiological function of CLL depending on the IgVH mutation status and may indicate that CLL may have a different response to BCR signaling inhibitors. Other in vitro studies have reported that the specific Btk inhibition of the agent PCI-32765 in the study produces more apoptosis and cytotoxicity in CLL cells than in normal B cells; and is induced by anti-apoptotic microenvironmental signals. Apoptosis, reduced secretion of chemokines CCL3 and CCL4, and decreased chemotaxis toward the chemokines CXCL12 and CXCL13. A detailed study of the pathophysiological effects of Btk in the source and/or maintenance of WM has not been reported. However, a recent report investigating the transgenic mouse model shows that constitutively active Btk expression causes selective expansion or survival of B-1 cells, driving germinal center-independent plasma cell differentiation, such as IgM+ in the spleen and bone marrow. Increased plasma cell numbers and significantly elevated serum IgM were demonstrated. Anti-nucleosome autoantibodies and glomerular IgM deposition were also observed. However, a sequence analysis study of 19 WM patients with low gamma globulinemia G and/or A failed to find any novel variants of the Btk promoter, flanking intron or exon.

Allogeneic stem cell transplantation is the only treatment that may cure CLL, but 70% of patients are diagnosed 65 years old, suffering from a combined disease that limits the applicability of the therapy, and exhibits an extended natural history in the presence or absence of specific treatment. The actual prognosis of CLL is variable and depends primarily on the clinical stage and certain genetic and molecular characteristics. The Rai and Binet clinical staging systems are able to range from the most advanced (thrombocytopenia) from 19 months to the earliest (no adenosis, visceral enlargement or defined anemia/thrombocytopenia in blood and bone marrow lymphocytes) The 150-month median OS differentiated the patient prognosis group. Additional prognostic differentiation was added for clinical staging by the presence or absence of IgVH and by profiling of inferior fluorescence in situ hybridization (iFISH) analysis of acquired chromosomal abnormalities, with unmutated IgVH and del(11q) and del ( 17p) Cytogenetics predicts poorer results.

CLL treatment strategies are complex and require first treatment (eg, such as fatigue or night sweats; large adenopathy/visceral enlargement; symptoms of progressive anemia/thrombocytopenia); and second choice of treatment options, generally involving one or more of the following: Purine nucleoside (fludarabine), alkylating agent (cyclophosphamide, chlorambucil, bendamustine), corticosteroids, anti-CD20 monoclonal antibody (rituximab) /ofolimumab) or anti-CD52 monoclonal antibody (alemtuzumab). The choice of specific therapy depends on the patient's age, disease patterns (eg, primarily node and non-node), expected drug resistance and contraindications, and the presence or absence of adverse prognostic features (such as del(11q) or del(17p)) And set. Although there are many therapies, the treatment options are ultimately limited by drug toxicity and tolerance, and patients who have not died of other diseases endure the progressive complications associated with cytopenia, lymphadenopathy and visceral enlargement, systemic Symptoms and infectious complications. Given the ageing characteristics of patient populations, oral and well tolerated treatments that exploit the new weaknesses of CLL should be welcome.

The basic principle of targeting Btk in CLL and SLL and its combination with lenalidomide

Strategies that specifically target B cells (eg, B cell depletion of the anti-CD20 monoclonal antibody rituximab and orfarizumab) have shown clinical efficacy in B cell lymphoma and CLL. Spleen tyrosine kinase (Syk) is a kinase close to Btk in the BCR signaling pathway. The use of the orally available Syk inhibitor fostamatinib disodium inhibits the clinical response of Syk in DLBCL, CLL and mantle cell lymphoma. Most convincingly, the clinical evidence for the concept of Btk inhibition has been shown by clinical investigation of the orally available Btk inhibitor PCI-32765, which has been reported to have DLBCL; mantle cells, marginal/mucosa-associated lymphoid tissue (MALT) and follicular lymphoma (FL), Waldenstrom macroglobulin Objective anti-tumor response and good tolerance of patients with blood (Waldenstrom's macroglobulinemia, WM) and CLL/SLL.

Therefore, based on the critical importance of Btk-mediated BCR signaling for the survival and proliferation of various malignant B cells; restricted cell expression of Btk in B cells, macrophages and monocytes; and the shown inhibition of Btk production Preclinical and early clinical evidence for the concept of anti-lymphoma, CLL and WM effects and acceptable clinical tolerance, targeting Btk with selective Btk inhibitors is a promising therapeutic strategy for further clinical investigations . Compound 1 (in its besylate form) has been shown to be safe as a single therapeutic agent and effective against CLL in the latest studies. As of September 11, 2012, 35 of 43 CLL patients had stable disease and continued to use N- (5-fluoro-2-(4-(2-methoxyethoxy)aniline) Monopyridyl-4-pyrimidinyl)phenyl)propenylamine besylate monotherapy. The lymph node size of 15 of the 23 patients has decreased, and the absolute lymphocyte count (ALC) of 28 of the 33 patients has increased early. See U.S. Patent Application Serial No. 13/661,678, the disclosure of which is hereby incorporated by reference in its entirety in its entirety in the entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire all all all all all This data strongly supports the use of BTK inhibitors and in particular Compound 1 to treat CLL. Compound 1 as a single agent is generally well tolerated up to 750 mg PO QD and has not yet reached the maximum tolerated dose (MTD). Additional doses that are still under investigation and currently under investigation include: 1000 mg QD, 1250 mg QD, 375 mg BID, and 500 mg BID. Lenalidomide has also been shown to exhibit good activity against patients with relapsed/refractory CLL.

Rituximab has also been shown to exhibit good activity against patients with relapsed/refractory CLL. In one study, rituximab, fludarabine, and cyclophosphamide were evaluated in 408 CLL patients and demonstrated a 86% response rate as observed with fludarabine/cyclophosphamide alone. Compared to the 73% response rate. The median progression-free survival was 39.8 months compared to 31.5 months observed for fludarabine/cyclophosphamide alone.

Thus, in some embodiments, the invention contemplates the use of a combination of a BTK inhibitor such as Compound 1 and lenalidomide for the treatment of CLL and SLL. In some embodiments, the invention contemplates the use of a BTK inhibitor such as Compound 1 in combination with lenalidomide and rituximab for the treatment of CLL and SLL. Compound 1 can be found to be effective as a single agent or in combination for patients with CLL, including but not limited to patients who have demonstrated one or more of the following prognosis/genetic markers and cytogenetic risk factors: chromosome 11q , 17p or 13q deletion, or 12th trisomy 13 (Trisomy 12) and 14q versus trisomy, ζ chain associated protein kinase 70 (ZAP 70) or immunoglobulin heavy chain variable region (IgVH ) No mutation.

In some embodiments, the invention encompasses the recognition that lenalidomide and a Btk inhibitor are synergistic in a diffuse large B-cell lymphoma (DLBCL) model, perhaps due to the two agents acting against IκB kinase (IKK) and downstream Concomitant inhibition of nuclear factor kappa B (NFkB) signaling. Accordingly, the present invention provides a method of treating, stabilizing or ameliorating the severity or progression of one or more BTK-associated diseases and conditions, comprising administering a combination of Compound 1 and lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering rituximab to a patient in need thereof. In some embodiments, the methods provided comprise administering to each of the patients in need, Compound 1 , lenalidomide, and rituximab.

III. Methods of treating diseases or disorders associated with BTK

In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of a disease or disorder selected from the group consisting of chronic lymphocytic leukemia and small lymphocytic lymphoma, the method comprising The patient is administered a combination of Compound 1 and lenalidomide. In some embodiments, the methods provided additionally comprise administering rituximab to a patient in need thereof. In some embodiments, the methods provided comprise administering to each of the patients in need, Compound 1 , lenalidomide, and rituximab.

In some embodiments, the invention provides a combination of Compound 1 and lenalidomide, wherein the combination exhibits a synergistic effect. In some embodiments, the methods provided comprise administering a combination of Compound 1 and lenalidomide to a patient in need thereof, wherein the amount of Compound 1 and/or lenalidomide administered is less than in each of the treatment regimens The amount of one or both compounds administered at the time of administration. In some embodiments, the methods provided additionally comprise administering rituximab to a patient in need thereof. In some embodiments, a method is provided comprising administering to a patient in need thereof each of the therapeutic agents Compound 1 , Lenalidomide, and Rituximab, wherein the therapeutic agents exhibit a synergistic effect.

In some embodiments, the methods provided comprise administering to a patient in need, a combination comprising a composition comprising Compound 1 and a composition comprising lenalidomide. In some embodiments, the methods provided additionally comprise administering to a patient in need thereof a composition comprising rituximab. In some embodiments, a method is provided comprising administering to a patient in need thereof a composition comprising each of Compound 1 , Lenalidomide, and Rituximab.

In some embodiments, the composition comprising Compound 1 additionally comprises one or more pharmaceutically acceptable excipients. In some of these embodiments, the composition comprising Compound 1 is formulated into an oral dosage form. In some embodiments, the oral dosage form is a capsule.

In some embodiments, the composition comprising lenalidomide additionally comprises one or more pharmaceutically acceptable excipients. In some of these embodiments, the composition comprising lenalidomide is formulated into an oral dosage form. In some embodiments, the oral dosage form is a capsule.

In some embodiments, the compound 1 in combination with an amine of lenalidomide in combination on a single pharmaceutically acceptable composition is administered.

In some embodiments, a method is provided comprising administering a unit dose of Compound 1 to a unit dose of lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering a unit dose of rituximab to a patient in need thereof. In some embodiments, the methods provided comprise administering a unit dose of Compound 1 , a unit dose of lenalidomide, and a unit dose of rituximab to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering a unit dose of rituximab to a patient in need thereof. In some embodiments, the unit dosage of Compound 1 is 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, or about 250 mg. In some embodiments, the unit dose of lenalidomide is about 2.5 mg, about 5 mg, about 7.5 mg, about 10 mg, about 15 mg, or about 25 mg.

In some embodiments, a method is provided comprising administering to a patient in need thereof a pharmaceutical composition comprising rituximab, wherein rituximab is administered as an infusion at a rate of 50 mg/h. In some embodiments, the infusion rate of rituximab is increased by 50 mg/h every 30 minutes to a maximum of 400 mg/h. In some embodiments, the infusion rate of rituximab is increased by 100 mg/h every 30 minutes to a maximum of 400 mg/h. Thus, in some embodiments, the infusion rate of rituximab is 100 mg/h. In some embodiments, the infusion rate of rituximab is 150 mg/h. In some embodiments, the infusion rate of rituximab is 200 mg/h. In some embodiments, the infusion rate of rituximab is 250 mg/h. In some embodiments, the infusion rate of rituximab is 300 mg/h. In some embodiments, the infusion rate of rituximab is 350 mg/h. In some embodiments, the infusion rate of rituximab is 400 mg/h.

In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of a disease or disorder selected from the group consisting of chronic lymphocytic leukemia and small lymphocytic lymphoma, the method comprising The patient is administered a combination of Compound 1 and lenalidomide, wherein the patient has failed at least one prior therapy. In some embodiments, a method is provided comprising administering to a patient in need thereof each of Compound 1 , lenalidomide, and rituximab, wherein the patient has failed at least one prior therapy.

In some embodiments, the methods provided comprise administering to a patient in need thereof a combination of from about 250 mg to about 1250 mg of Compound 1 and from about 2.5 mg to about 25 mg of lenalidomide. In some embodiments, the methods provided comprise administering to a patient in need thereof a combination of from about 500 mg to about 1250 mg of Compound 1 and from about 2.5 mg to about 25 mg of lenalidomide. In some embodiments, the methods provided comprise administering from about 250 mg to about 1250 mg of Compound 1 , from about 2.5 mg to about 25 mg of lenalidomide, and from about 375 mg/m ² to about 500 mg/m ² of lysine to a patient in need thereof. Waximab.

In some embodiments, the methods provided comprise administering from about 750 mg to about 1000 mg of Compound 1 and from about 2.5 mg to about 10 mg of lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering from about 750 mg to about 1000 mg of Compound 1 and from about 2.5 mg to about 15 mg of lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering from about 750 mg to about 1000 mg of Compound 1 and from about 5 mg to about 10 mg of lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering from about 750 mg to about 1000 mg of Compound 1 and from about 5 mg to about 15 mg of lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering from about 750 mg to about 1000 mg of Compound 1 and from about 7.5 mg to about 10 mg of lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering from about 750 mg to about 1000 mg of Compound 1 and from about 7.5 mg to about 15 mg of lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering from about 750 mg to about 1000 mg of Compound 1 and from about 10 mg to about 15 mg of lenalidomide to a patient in need thereof. In some embodiments, the method further comprising providing to a patient in need there of administration and about 375mg / m ² to about 500mg / m ² rituximab.

In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of a disease or disorder selected from the group consisting of chronic lymphocytic leukemia and small lymphocytic lymphoma, the method comprising The patient is administered a combination of about 125 mg BID to about 500 mg BID Compound 1 and about 2.5 mg QD to about 15 mg QD lenalidomide. In some embodiments, the method further comprising providing to a patient in need there of administration and about 375mg / m ² to about 500mg / m ² rituximab. In some embodiments, the methods provided comprise administering to a patient in need thereof about 125 mg BID to about 500 mg BID Compound 1 , about 2.5 mg QD to about 15 mg QD lenalidomide, and about 375 mg/m ² to about 500 mg/ m ² rituximab.

In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of a disease or disorder selected from the group consisting of chronic lymphocytic leukemia and small lymphocytic lymphoma, the method comprising The patient is administered a combination of about 125 mg BID to about 500 mg BID Compound 1 and about 5 mg QD to about 15 mg QD lenalidomide. In some embodiments, the method further comprising providing to a patient in need there of administration and about 375mg / m ² to about 500mg / m ² rituximab. In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of a disease or disorder selected from the group consisting of chronic lymphocytic leukemia and small lymphocytic lymphoma, the method comprising the patient administered from about 125mg BID and 500mg BID compound to about 1, to about 2.5mg QD about 15mg QD lenalidomide and about 375mg / m ² to about 500mg / m ² rituximab.

In some embodiments, the methods provided comprise administering about 125 mg BID to about 500 mg BID Compound 1 and about 2.5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 500 mg BID Compound 1 and about 2.5 mg QD to about 10 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 500 mg BID Compound 1 and about 2.5 mg QD to about 7.5 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 500 mg BID Compound 1 and about 2.5 mg QD to about 5 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering to a patient in need thereof about 125 mg BID to about 500 mg BID Compound 1 and about 5 mg QD to about 15 mg QD lenalidomide. In some embodiments, the methods provided comprise administering about 125 mg BID to about 500 mg BID Compound 1 and about 5 mg QD to about 10 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 500 mg BID Compound 1 and about 5 mg QD to about 7.5 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 500 mg BID Compound 1 and about 7.5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 500 mg BID Compound 1 and about 7.5 mg QD to about 10 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 500 mg BID Compound 1 and about 10 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the method further comprising providing to a patient in need there of administration and about 375mg / m ² to about 500mg / m ² rituximab.

In some embodiments, the methods provided comprise administering about 125 mg BID to about 375 mg BID Compound 1 and about 2.5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 375 mg BID Compound 1 and about 2.5 mg QD to about 10 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 375 mg BID Compound 1 and about 2.5 mg QD to about 7.5 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 375 mg BID Compound 1 and about 2.5 mg QD to about 5 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering to a patient in need thereof about 125 mg BID to about 375 mg BID Compound 1 and about 5 mg QD to about 15 mg QD lenalidomide. In some embodiments, the methods provided comprise administering about 125 mg BID to about 375 mg BID Compound 1 and about 5 mg QD to about 10 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 375 mg BID Compound 1 and about 5 mg QD to about 7.5 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 375 mg BID Compound 1 and about 7.5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 375 mg BID Compound 1 and about 7.5 mg QD to about 10 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 375 mg BID Compound 1 and about 10 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the method further comprising providing to a patient in need there of administration and about 375mg / m ² to about 500mg / m ² rituximab.

In some embodiments, the methods provided comprise administering about 125 mg BID to about 250 mg BID Compound 1 and about 2.5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 125 mg BID to about 250 mg BID Compound 1 and about 2.5 mg QD to about 10 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering to a patient in need thereof about 125 mg BID to about 250 mg BID Compound 1 and about 2.5 mg QD to about 7.5 mg QD lenalidomide. In some embodiments, the methods provided comprise administering to a patient in need thereof about 125 mg BID to about 250 mg BID Compound 1 and about 2.5 mg QD to about 5 mg QD lenalidomide. In some embodiments, the methods provided comprise administering about 125 mg BID to about 250 mg BID Compound 1 and about 5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering to a patient in need thereof about 125 mg BID to about 250 mg BID Compound 1 and about 5 mg QD to about 10 mg QD lenalidomide. In some embodiments, the methods provided comprise administering about 125 mg BID to about 250 mg BID Compound 1 and about 5 mg QD to about 7.5 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering to a patient in need thereof about 125 mg BID to about 250 mg BID Compound 1 and about 7.5 mg QD to about 15 mg QD lenalidomide. In some embodiments, the methods provided comprise administering to a patient in need thereof about 125 mg BID to about 250 mg BID Compound 1 and about 7.5 mg QD to about 10 mg QD lenalidomide. In some embodiments, the methods provided comprise administering to a patient in need thereof about 125 mg BID to about 250 mg BID Compound 1 and about 10 mg QD to about 15 mg QD lenalidomide. In some embodiments, the method further comprising providing to a patient in need there of administration and about 375mg / m ² to about 500mg / m ² rituximab.

In some embodiments, the methods provided comprise administering about 250 mg BID to about 500 mg BID Compound 1 and about 2.5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 250 mg BID to about 500 mg BID Compound 1 and about 2.5 mg QD to about 10 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 250 mg BID to about 500 mg BID Compound 1 and about 2.5 mg QD to about 7.5 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering to a patient in need thereof about 250 mg BID to about 500 mg BID Compound 1 and about 2.5 mg QD to about 5 mg QD lenalidomide. In some embodiments, the methods provided comprise administering about 250 mg BID to about 500 mg BID Compound 1 and about 5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 250 mg BID to about 500 mg BID Compound 1 and about 5 mg QD to about 10 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering to a patient in need thereof about 250 mg BID to about 500 mg BID Compound 1 and about 5 mg QD to about 7.5 mg QD lenalidomide. In some embodiments, the methods provided comprise administering about 250 mg BID to about 500 mg BID Compound 1 and about 7.5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 250 mg BID to about 500 mg BID Compound 1 and about 7.5 mg QD to about 10 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 250 mg BID to about 500 mg BID Compound 1 and about 10 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the method further comprising providing to a patient in need there of administration and about 375mg / m ² to about 500mg / m ² rituximab.

In some embodiments, the methods provided comprise administering about 250 mg BID to about 375 mg BID Compound 1 and about 2.5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 250 mg BID to about 375 mg BID Compound 1 and about 2.5 mg QD to about 10 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 250 mg BID to about 375 mg BID Compound 1 and about 2.5 mg QD to about 7.5 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 250 mg BID to about 375 mg BID Compound 1 and about 2.5 mg QD to about 5 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 250 mg BID to about 375 mg BID Compound 1 and about 5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering to a patient in need thereof about 250 mg BID to about 375 mg BID Compound 1 and about 5 mg QD to about 10 mg QD lenalidomide. In some embodiments, the methods provided comprise administering about 250 mg BID to about 375 mg BID Compound 1 and about 5 mg QD to about 7.5 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 250 mg BID to about 375 mg BID Compound 1 and about 7.5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 250 mg BID to about 375 mg BID Compound 1 and about 7.5 mg QD to about 10 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering to a patient in need thereof about 250 mg BID to about 375 mg BID Compound 1 and about 10 mg QD to about 15 mg QD lenalidomide. In some embodiments, the method further comprising providing to a patient in need there of administration and about 375mg / m ² to about 500mg / m ² rituximab.

In some embodiments, the methods provided comprise administering about 375 mg BID to about 500 mg BID Compound 1 and about 2.5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 375 mg BID to about 500 mg BID Compound 1 and about 2.5 mg QD to about 10 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 375 mg BID to about 500 mg BID Compound 1 and about 2.5 mg QD to about 7.5 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 375 mg BID to about 500 mg BID Compound 1 and about 2.5 mg QD to about 5 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 375 mg BID to about 500 mg BID Compound 1 and about 5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering from a patient in need thereof about 375 mg BID to about 500 mg BID Compound 1 and about 5 mg QD to about 10 mg QD lenalidomide. In some embodiments, the methods provided comprise administering about 375 mg BID to about 500 mg BID Compound 1 and about 5 mg QD to about 7.5 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering about 375 mg BID to about 500 mg BID Compound 1 and about 7.5 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided comprise administering from a patient in need thereof about 375 mg BID to about 500 mg BID Compound 1 and about 7.5 mg QD to about 10 mg QD lenalidomide. In some embodiments, the methods provided comprise administering about 375 mg BID to about 500 mg BID Compound 1 and about 10 mg QD to about 15 mg QD lenalidomide to a patient in need thereof. In some embodiments, the method further comprising providing to a patient in need there of administration and about 375mg / m ² to about 500mg / m ² rituximab.

In some embodiments, the methods provided comprise administering about 125 mg of BID Compound 1 and about 2.5 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 125 mg of BID Compound 1 and about 5 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 125 mg of BID Compound 1 and about 7.5 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the method provided further comprises administering to a patient in need of and about 375mg / m ² or from about 500mg / m ² rituximab.

In some embodiments, the methods provided comprise administering about 125 mg of BID Compound 1 and about 10 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 125 mg of BID Compound 1 and about 15 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 250 mg of BID Compound 1 and about 2.5 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 250 mg of BID Compound 1 and about 5 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 250 mg of BID Compound 1 and about 7.5 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 250 mg of BID Compound 1 and about 10 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 250 mg of BID Compound 1 and about 15 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 375 mg of BID Compound 1 and about 2.5 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 375 mg of BID Compound 1 and about 5 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 375 mg of BID Compound 1 and about 7.5 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 375 mg of BID Compound 1 and about 10 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 375 mg of BID Compound 1 and about 15 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 500 mg of BID Compound 1 and about 2.5 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 500 mg of BID Compound 1 and about 5 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 500 mg of BID Compound 1 and about 7.5 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 500 mg of BID Compound 1 and about 10 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the methods provided comprise administering about 500 mg of BID Compound 1 and about 15 mg of QD lenalidomide to a patient in need thereof. In some embodiments, the methods provided additionally comprise administering about 375 mg/m ² or about 500 mg/m ² of rituximab to a patient in need thereof.

In some embodiments, the combination of Compound 1 and lenalidomide is administered over a period of 28 consecutive days ("28 day cycle"). In some embodiments, a combination of Compound 1 and lenalidomide is administered for two, three, four, five or six 28 day cycles. In some embodiments, a combination of Compound 1 and lenalidomide is administered for seven, eight, nine, ten, eleven, twelve or twelve or more 28 day cycles. In some embodiments, the combination of Compound 1 and lenalidomide is administered once a day for at least thirteen, at least fourteen, at least fifteen, at least sixteen, at least seventeen, at least eighteen At least nineteen or at least twenty 28-day cycles. In some embodiments, a combination of Compound 1 and lenalidomide is administered to a patient for a lifetime. In some embodiments, the patient is administered a combination of Compound 1 and lenalidomide for one or more 28-day cycles, and the patient is additionally administered one or more of Compound 1 or Lenalidomide. A 28-day cycle. In some embodiments, the methods provided additionally comprise administering rituximab to a patient in need on the first or second day of the first cycle. In some embodiments, the method provided additionally comprises administering rituximab to a patient in need on the first, second, fourth, fifth, and sixth cycles. In some embodiments, the methods provided comprise administering to a patient in need thereof, each of Compound 1 , Lenalidomide, and Rituximab, wherein Compound 1 and lenalidomide are administered to the patient. Each of the one or more 28-day cycles and the first day, the second cycle, the third cycle, the fourth cycle, the fifth cycle, and/or the first or second day of the sixth cycle Waximab.

In some embodiments, two adjacent 28 day periods may be separated by a rest period. Such rest periods may be one day, two days, three days, four days, five days, six days, seven days or more, during which no one of Compound 1 and lenalidomide is administered to the patient or Both. In a preferred embodiment, two adjacent 28 day periods are continuous.

In some embodiments, a method is provided comprising administering to a patient in need thereof a combination of Compound 1 and lenalidomide, wherein the patient has failed at least one prior therapy.

In some embodiments, the invention provides a system for treating, stabilizing or attenuating the severity of one or more diseases or conditions associated with BTK, the system comprising Compound 1 and lenalidomide. In some embodiments, the system is a kit. In some such embodiments, the kit comprises: a pharmaceutical composition comprising Compound 1 and a pharmaceutical composition comprising lenalidomide. In some embodiments, the kit comprises one or more unit doses of Compound 1 in combination with one or more unit doses of lenalidomide. In some embodiments, the kit comprising: a pharmaceutical composition comprising a compound of 1, lenalidomide comprising the pharmaceutical composition comprises rituximab and pharmaceutical compositions of infliximab. In some embodiments, the kit comprises one or more unit doses of Compound 1 , one or more unit doses of lenalidomide, and one or more unit doses of rituximab.

In some embodiments, the kit comprises two 125 mg doses of Compound 1 and a 2.5 mg dose of lenalidomide. In some embodiments, the kit comprises two 125 mg doses of Compound 1 and a 5 mg dose of lenalidomide. In some embodiments, the kit comprises two 125 mg doses of Compound 1 and a 7.5 mg dose of lenalidomide. In some embodiments, the kit comprises two 125 mg doses of Compound 1 and one 10 mg dose of lenalidomide. In some embodiments, the kit comprises two 125 mg doses of Compound 1 and a 15 mg dose of lenalidomide.

In some embodiments, the kit comprises two 250 mg doses of Compound 1 and a 2.5 mg dose of lenalidomide. In some embodiments, the kit comprises two 250 mg doses of Compound 1 and a 5 mg dose of lenalidomide. In some embodiments, the kit comprises two 250 mg doses of Compound 1 and a 7.5 mg dose of lenalidomide. In some embodiments, the kit comprises two 250 mg doses of Compound 1 and a single 10 mg dose of lenalidomide. In some embodiments, the kit comprises two 250 mg doses of Compound 1 and a 15 mg dose of lenalidomide.

In some embodiments, the kit comprises a dose of 375mg twice a 2.5mg dose of compound 1 and of lenalidomide. In some embodiments, the kit comprises two 375 mg doses of Compound 1 and a single 5 mg dose of lenalidomide. In some embodiments, the kit comprises two 375 mg doses of Compound 1 and a 7.5 mg dose of lenalidomide. In some embodiments, the kit comprises two 375 mg doses of Compound 1 and a single 10 mg dose of lenalidomide. In some embodiments, the kit comprises two 375 mg doses of Compound 1 and a 15 mg dose of lenalidomide.

In some embodiments, the kit comprises two 500 mg doses of Compound 1 and one 2.5 mg dose of lenalidomide. In some embodiments, the kit comprises two 500 mg doses of Compound 1 and one 5 mg dose of lenalidomide. In some embodiments, the kit comprises two 500 mg doses of Compound 1 and a 7.5 mg dose of lenalidomide. In some embodiments, the kit comprises two 500 mg doses of Compound 1 and one 10 mg dose of lenalidomide. In some embodiments, the kit comprises two 500 mg doses of Compound 1 and a 15 mg dose of lenalidomide.

In some embodiments, the kit comprises seven (7) daily doses of Compound 1 and lenalidomide. In some of these embodiments, the kit comprises fourteen (14) doses of 125 mg of Compound 1 and seven (7) doses of 2.5 mg of lenalidomide. In some of these embodiments, the kit comprises fourteen (14) doses of 125 mg of Compound 1 and seven (7) doses of 5 mg of lenalidomide. In some of these embodiments, the kit comprises fourteen (14) doses of 125 mg of Compound 1 and seven (7) doses of 7.5 mg of lenalidomide. In some of these embodiments, the kit comprises fourteen (14) doses of 125 mg of Compound 1 and seven (7) doses of 10 mg of lenalidomide. In some of these embodiments, the kit comprises fourteen (14) doses of 125 mg of Compound 1 and seven (7) doses of 15 mg of lenalidomide.

In some of these embodiments, the kit comprises fourteen (14) 250 mg doses of Compound 1 and seven (7) 2.5 mg doses of lenalidomide. In some of these embodiments, the kit comprises fourteen (14) 250 mg doses of Compound 1 and seven (7) 5 mg doses of lenalidomide. In some of these embodiments, the kit comprises fourteen (14) 250 mg doses of Compound 1 and seven (7) 7.5 mg doses of lenalidomide. In some of these embodiments, the kit comprises fourteen (14) 250 mg doses of Compound 1 and seven (7) doses of 10 mg dose of lenalidomide. In some of these embodiments, the kit comprises fourteen (14) 250 mg doses of Compound 1 and seven (7) 15 mg doses of lenalidomide.

In some of these embodiments, the kit comprises fourteen (14) 375 mg doses of Compound 1 and seven (7) 2.5 mg doses of lenalidomide. In some of these embodiments, the kit comprises fourteen (14) 375 mg doses of Compound 1 and seven (7) 5 mg doses of lenalidomide. In some of these embodiments, the kit comprises fourteen (14) 375 mg doses of Compound 1 and seven (7) 7.5 mg doses of lenalidomide. In some such embodiments, the kit comprises fourteen (14) times a 375 mg dose of Compound 1 and seven (7) times a 10 mg dose of lenalidomide. In some such embodiments, the kit comprises fourteen (14) 375 mg doses of Compound 1 and seven (7) 15 mg doses of lenalidomide.

In some embodiments, the kit comprises fourteen (14) times a 500 mg dose of Compound 1 and seven (7) times a 2.5 mg dose of lenalidomide. In some embodiments, the kit comprises fourteen (14) times a 500 mg dose of Compound 1 and seven (7) times a 5 mg dose of lenalidomide. In some embodiments, the kit comprises fourteen (14) times a 500 mg dose of Compound 1 and seven (7) times a 7.5 mg dose of lenalidomide. In some embodiments, the kit comprises fourteen (14) times a 500 mg dose of Compound 1 and seven (7) times a 10 mg dose of lenalidomide. In some embodiments, the kit comprises fourteen (14) times a 500 mg dose of Compound 1 and seven (7) times a 15 mg dose of lenalidomide.

In some embodiments, the kit comprises 28 daily doses of Compound 1 and lenalidomide. In some embodiments, the kit comprises fifty-six (56) doses of 125 mg of Compound 1 and twenty-eight (28) doses of 2.5 mg of lenalidomide. In some embodiments, the kit comprises fifty-six (56) doses of 125 mg of Compound 1 and twenty-eight (28) doses of 5 mg of lenalidomide. In some embodiments, the kit comprises fifty-six (56) doses of 125 mg of Compound 1 and twenty-eight (28) doses of 7.5 mg of lenalidomide. In some embodiments, the kit comprises fifty-six (56) doses of 125 mg of Compound 1 and twenty-eight (28) doses of 10 mg of lenalidomide. In some embodiments, the kit comprises fifty-six (56) doses of 125 mg of Compound 1 and twenty-eight (28) doses of 15 mg of lenalidomide. In some embodiments, the kit additionally comprises a 100 mg/10 mL vial of rituximab. In some embodiments, the kit additionally comprises a 500 mg/50 mL vial of rituximab.

In some embodiments, the kit comprises fifty-six (56) 250 mg doses of Compound 1 and twenty-eight (28) doses of 2.5 mg dose of lenalidomide. In some embodiments, the kit comprises fifty-six (56) 250 mg doses of Compound 1 and twenty-eight (28) doses of 5 mg dose of lenalidomide. In some embodiments, the kit comprises fifty-six (56) 250 mg doses of Compound 1 and twenty-eight (28) doses of 7.5 mg dose of lenalidomide. In some embodiments, the kit comprises fifty-six (56) 250 mg doses of Compound 1 and twenty-eight (28) doses of 10 mg dose of lenalidomide. In some embodiments, the kit comprises fifty-six (56) 250 mg doses of Compound 1 and twenty-eight (28) doses of 15 mg dose of lenalidomide. In some embodiments, the kit additionally comprises a 100 mg/10 mL vial of rituximab. In some embodiments, the kit additionally comprises a 500 mg/50 mL vial of rituximab.

In some embodiments, the kit comprises fifty-six (56) 375 mg doses of Compound 1 and twenty-eight (28) doses of 2.5 mg dose of lenalidomide. In some embodiments, the kit comprises fifty-six (56) times a 375 mg dose of Compound 1 and twenty-eight (28) doses of a 5 mg dose of lenalidomide. In some embodiments, the kit comprises fifty-six (56) 375 mg doses of Compound 1 and twenty-eight (28) 7.5 mg doses of lenalidomide. In some embodiments, the kit comprises fifty-six (56) 375 mg doses of Compound 1 and twenty-eight (28) doses of 10 mg dose of lenalidomide. In some embodiments, the kit comprises fifty-six (56) 375 mg doses of Compound 1 and twenty-eight (28) doses of 15 mg dose of lenalidomide. In some embodiments, the kit additionally comprises a 100 mg/10 mL vial of rituximab. In some embodiments, the kit additionally comprises a 500 mg/50 mL vial of rituximab.

In some embodiments, the kit comprises fifty-six (56) doses of 500 mg of Compound 1 and twenty-eight (28) doses of 2.5 mg of lenalidomide. In some embodiments, the kit comprises fifty-six (56) times a 500 mg dose of Compound 1 and twenty-eight (28) doses of a 5 mg dose of lenalidomide. In some embodiments, the kit comprises fifty-six (56) times a 500 mg dose of Compound 1 and twenty-eight (28) doses of a 7.5 mg dose of lenalidomide. In some embodiments, the kit comprises fifty-six (56) times a 500 mg dose of Compound 1 and twenty-eight (28) doses of a 10 mg dose of lenalidomide. In some embodiments, the kit comprises fifty-six (56) 500 mg doses of Compound 1 and twenty-eight (28) doses of 15 mg dose of lenalidomide. In some embodiments, the kit additionally comprises a 100 mg/10 mL vial of rituximab. In some embodiments, the kit additionally comprises a 500 mg/50 mL vial of rituximab.

IV. Formulation comprising Compound 1

As indicated above, the method provided comprises administering to a patient in need thereof a pharmaceutically acceptable composition comprising Compound 1 , wherein the pharmaceutically acceptable composition is an oral dosage form. In some embodiments, the pharmaceutically acceptable composition is formulated into a capsule.

In certain embodiments, the methods provided comprise administering to a patient in need thereof a pharmaceutically acceptable composition comprising Compound 1 and one or more pharmaceutically acceptable excipients, such as a binder. Agents, film coats, diluents, disintegrants, surfactants (wetting agents), lubricants and slip agents (adsorbents) or combinations thereof. It will be readily understood by those skilled in the art that the enumeration of specific components is not intended to be limiting; in some cases, a particular component may suitably be adapted to more than one category. Likewise, it is to be understood that in the case of a particular formulation, such as depending on the amount of ingredient and/or the presence of other ingredients and/or active compound, the same component may sometimes perform different functions or perform more than one function. In some embodiments, the pharmaceutically acceptable composition is a blended powder.

i. Adhesives and thinners

The pharmaceutical compositions used in the present invention may comprise one or more binders. Binders are used in the formulation of solid oral dosage forms to keep the active pharmaceutical ingredients in combination with the inactive ingredients. In some embodiments, the pharmaceutical compositions of the present invention comprise from about 5% to about 50% (w/w) of one or more binders and/or diluents. In some embodiments, the pharmaceutical compositions of the present invention comprise about 20% (w/w) one or more binders and/or diluents. Suitable binders and/or diluents (also known as "fillers") are known in the art. Representative binders and/or diluents include, but are not limited to, starches such as cellulose (low molecular weight HPC (hydroxypropyl cellulose), microcrystalline cellulose (eg Avicel ^® ), low molecular weight HPMC (hydroxylpropyl) Cellulose), low molecular weight carboxymethylcellulose, ethylcellulose); sugars such as lactose (ie, lactose monohydrate), sucrose, dextrose, fructose, maltose, glucose; and polyols such as sorbose Alcohol, mannitol, lactitol, maltitol, and xylitol, or a combination thereof. In some embodiments, the provided compositions comprise a binder of microcrystalline cellulose and/or monohydrated lactose.

Ii. disintegrant

The pharmaceutical composition used in the present invention may additionally comprise one or more disintegrants. Suitable disintegrants are known in the art and include, but are not limited to, agar, calcium carbonate, sodium carbonate, sodium bicarbonate, cross-linked sodium carboxymethyl cellulose (crosslinking) Sodium carboxymethylcellulose (croscarmellose sodium), sodium carboxymethyl starch (sodium starch glycolate), microcrystalline cellulose or a combination thereof. In some embodiments, the formulation provided comprises from about 1% to about 25% of a disintegrant, based on the total weight of the formulation.

Iii. Surfactant

Surfactants (also known as bioavailability enhancers) are well known in the art and are Drug release and absorption are often promoted by enhancing the solubility of poorly soluble drugs. Representative surfactants include, but are not limited to, poloxamers, polyoxyethylene ethers, polyoxyethylene fatty acid esters, polyethylene glycol fatty acid esters, polyoxyethylene hydrogenated castor oil, polyoxyethylene alkylenes. Ether ethers, polysorbates, and combinations thereof. In certain embodiments, the surfactant is a poloxamer. In some of these embodiments, the poloxamer is poloxamer 407. In some embodiments, the compositions used in the present invention comprise from about 1% to about 30% by weight, based on the total weight of the blended powder, of a surfactant.

Iv. Lubricant

The pharmaceutical compositions of the present invention may additionally comprise one or more lubricants. Lubricants are agents that are added in small amounts to the formulation to improve certain processing characteristics. The lubricant prevents the compounding mixture from sticking to the compression machine and enhances product flow by reducing interparticle friction. Representative lubricants include, but are not limited to, magnesium stearate, glyceride, sodium stearyl fumarate, and fatty acids (i.e., palmitic acid and stearic acid). In certain embodiments, the lubricant is magnesium stearate. In some embodiments, the provided formulation comprises from about 0.2% to about 3% lubricant, based on the total weight of the given formulation.

Slip agent

The pharmaceutical compositions of the present invention may additionally comprise one or more slip agents. Representative slip agents include, but are not limited to, ceria (i.e., fumed ceria), microcrystalline cellulose, starch (i.e., corn starch), and carbonates (i.e., calcium carbonate and magnesium carbonate). In some embodiments, the provided formulation comprises from about 0.2% to about 3% slipper, based on the total weight of the given formulation.

Vi. N-(3-(5-fluoro-2-(4-(2-methoxyethoxy)anilinyl)pyrimidin-4-ylamino)phenyl)acrylamidobenzenesulfonate

As described above, the present invention provides a method of treating a disease or disorder selected from the group consisting of chronic lymphocytic leukemia and small lymphocytic lymphoma, the method comprising administering a combination of Compound 1 and lenalidomide to a patient in need thereof. Benzenesulfonate compound 1 of N - (3- (5-fluoro-2- (4- (2-methoxyethoxy) phenylamino) pyrimidin-4-ylamino) phenyl) acrylamide benzene Sulfonates have recently been identified and are currently used as monotherapy in clinical trials for patients with relapsed or refractory B-cell non-Hodgkin's lymphoma (B-NHL), chronic lymphocytic leukemia (CLL) and Val Individuals of Dengstrom's macroglobulinemia (WM). Thus, in some embodiments, the method comprises providing to a patient in need of administration of a compound besylate.

In some embodiments, the methods provided comprise administering to a patient in need thereof a pharmaceutically acceptable composition comprising from about 5% to about 60% of Compound 1 based on the total weight of the formulation. Benzene sulfonate. In some embodiments, the methods provided comprise administering to a patient in need thereof a pharmaceutically acceptable composition comprising from about 5% to about 15%, or about the total weight of the composition. From 7% to about 15%, or from about 7% to about 10%, or from about 9% to about 12% of the besylate salt of Compound 1 . In some embodiments, the methods provided comprise administering to a patient in need thereof a pharmaceutically acceptable composition comprising from about 25% to about 75%, or about 30, based on the total weight of the formulation. % to about 60%, or from about 40% to about 50%, or from about 40% to about 45% of the besylate salt of Compound 1 . In certain embodiments, the methods provided comprise administering to a patient in need thereof a pharmaceutically acceptable composition comprising about 8%, based on the total weight of the given composition or formulation. 9%, about 10%, about 11%, about 12%, about 13%, about 20%, about 30%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45% About 50%, about 60%, about 70% or about 75% of the besylate salt of Compound 1 .

In some such embodiments, the methods provided comprise administering to a patient in need thereof a pharmaceutical composition comprising a unit dose of Compound 1 , wherein Compound 1 is in the form of a besylate salt. In some such embodiments, the unit dosage is an amount of the free base sufficient to provide 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, or about 250 mg of Compound 1 . . In some embodiments, the pharmaceutical composition comprising the besylate salt of Compound 1 is a solid oral dosage form.

In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of a disease or disorder selected from the group consisting of chronic lymphocytic leukemia and small lymphocytic lymphoma, the method comprising The patient is administered a combination of Compound 1 and lenalidomide, wherein Compound 1 is administered as a besylate salt. In some such embodiments, the besylate salt of Compound 1 is administered as a composition comprising one or more pharmaceutically acceptable excipients, one or more pharmaceutically acceptable excipients selected Self-adhesives, film coats, diluents, disintegrants, surfactants, lubricants and slip agents. In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of a disease or disorder selected from the group consisting of chronic lymphocytic leukemia and small lymphocytic lymphoma, the method comprising The patient is administered each of Compound 1 , lenalidomide and rituximab, wherein Compound 1 is administered as a besylate salt. In some such embodiments, the besylate salt of Compound 1 is administered as a composition comprising one or more pharmaceutically acceptable excipients, one or more pharmaceutically acceptable excipients selected Self-adhesives, film coats, diluents, disintegrants, surfactants, lubricants and slip agents.

In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of a disease or disorder selected from the group consisting of chronic lymphocytic leukemia and small lymphocytic lymphoma, the method comprising comprising administering to the patient a compound of the besylate salt of a pharmaceutical composition and to a combination of lenalidomide, wherein the amount of a compound of a besylate sufficient to transfer about 125mg, about 250mg, about 325 mg, about 375 mg of, about 400mg, About 500 mg, about 625 mg, about 750 mg, about 1000 mg, or about 1250 mg of the free base of Compound 1 . In some such embodiments, the pharmaceutical composition additionally comprises one or more pharmaceutically acceptable shapes selected from the group consisting of binders, film coats, diluents, disintegrants, surfactants, lubricants, and slip agents. Agent. In some such embodiments, the pharmaceutical composition comprises one or more medicinal materials selected from the group consisting of microcrystalline cellulose, lactose monohydrate, sodium starch, poloxamer 407, aerosolized cerium oxide, and magnesium stearate. Acceptable excipients. In some embodiments, the invention provides a method of treating, stabilizing or ameliorating the severity or progression of a disease or disorder selected from the group consisting of chronic lymphocytic leukemia and small lymphocytic lymphoma, the method comprising comprising administering to the patient a compound of the benzenesulfonate of 1 (i.e., compound 1 besylate), lenalidomide and each of rituximab in the pharmaceutical composition of 1 wherein the acid compound The amount of salt is sufficient to deliver about 125 mg, about 250 mg, about 325 mg, about 375 mg, about 400 mg, about 500 mg, about 625 mg, about 750 mg, about 1000 mg, or about 1250 mg of the free base of Compound 1 .

V. Process for preparing a pharmaceutical composition comprising Compound 1 Dry blending process:

Weighed N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl)propenylaminobenzenesulfonic acid Salt, ground microcrystalline cellulose, ground sodium starch glycolate, ground monohydrate lactose, ground poloxamer 407 and selected aerosolized cerium oxide and mechanically blended. The intragranular portion of the screened magnesium stearate (2.0%, according to Table 1 below) was added to the blender and the blend was blended. The blended formulation is then rolled, ground and subsequently blended. The blended formulation is rolled again, ground and subsequently blended. The remainder or extragranular portion of magnesium stearate (0.5% according to Table 1 below) was added and the final formulation was blended. The capsule is mechanically filled or manually filled via a flood filling method.

All of the features of the various aspects of the invention apply to all other aspects with the necessary modifications. The references (including but not limited to) patents, patent applications, and journal articles referred to herein are hereby incorporated by reference in their entirety as if they are fully incorporated.

In order to more fully understand the invention described herein, the following examples are set forth. It is understood that the examples are for illustrative purposes only and are not to be construed as limiting the invention in any way.

illustration Example 1 Dose escalation study

N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilinyl)pyrimidin-4-ylamino)phenyl)propenylamine benzenesulfonate is benzenesulfonate The acid form is developed by chemically synthesizing small molecules of substituted pyrimidines and is a white to off-white crystalline powder. N - (3- (5- fluoro-2- (4- (2-methoxyethoxy) phenylamino) pyrimidin-4-ylamino) phenyl) acrylamide benzenesulfonate of oral Btk Effective (IC ₅₀ <0.5 nM) and selective small molecule inhibitors. N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl)propenylamine besylate is shown in water A solubility of about 0.16 mg/mL and a maximum water solubility of 0.40 mg/mL at about pH 3.0. N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl)propenylamine benzenesulfonate in ethanol The solubility is about 10 mg/mL. N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl)acrylamidobenzenesulfonate did not show the need Specially treated for environmental instability (ie heat, acid, alkali).

Formulating N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilinyl)pyrimidin-4-ylamino)phenyl)acrylamidobenzenesulfonate to contain The components and amounts of capsules listed in Table 1 were used to obtain the study drug. The capsules listed in Table 1 will be administered during the dose escalation and expansion group study.

2.0% (8.30 mg) intragranular; 0.5% (2.08 mg) extragranular.

Lenalidomide is also known as 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione and has the following structure:

Lenalidomide was an off-white to pale yellow solid powder and was purchased from Celgene Corporation in the form of 2.5 mg, 5 mg, 10 mg, 15 mg, and 25 mg capsules. Each lenalidomide capsule contains anhydrous lactose, microcrystalline cellulose, croscarmellose sodium, and magnesium stearate.

In the future, lenalidomide will be formulated into capsules containing the components and amounts listed in Table 2 to obtain the study drug. The capsules listed in Table 2 will be administered according to the required dose during the dose escalation and expansion group study and represent the clinical supply form. Since commercially available products can also be used, the components and amounts of commercially available products are listed in Table 3.

^a Adjust the weight of lenalidomide for analysis and water content.

^b Adjust the weight of anhydrous lactose, NF/EP to achieve the target total blend weight of the batch.

^a Adjust the weight of lenalidomide for analysis and water content.

^b Adjust the weight of anhydrous lactose, NF/EP to achieve the target total blend weight of the batch.

^c capsule shells are supplied by Capsugel.

Research design

Non-tolerable doses (NTD) for individuals with relapsed or refractory CLL or SLL who have failed at least one prior treatment regimen in a "3+3" dose escalation and expansion study to determine the combination of Compound 1 and lenalidomide ), optimal biological effect dose (OBE) and maximum tolerated dose (MTD). Approximately 42 patients will be enrolled in the study.

Three (3) doses and three recommended doses are defined as outlined in Table 4.

* Suggest additional doses.

The study therapeutic is administered as scheduled at the prescribed dose over a 28 day period until the disease progresses, the toxicity is unacceptable, or is interrupted for any other reason. The individual will continue to start the dose until a preliminary recommended phase 2 dose (RP2D) is determined, at which point it can be converted to a preliminary RP2D.

Within each group, the individual used N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino) during the first 28 days of treatment) Combination of phenyl) acrylamide sulfonate and lenalidomide PO (oral) BID (twice daily) and assessment of safety, tolerability and DLT, and pharmacokinetics ("PK") , pharmacodynamics ("PD") and disease response. In some cases, a physician investigator may choose to rest the patient during the study, during which time the patient does not receive treatment. For example, a physician investigator may choose to rest the patient due to the occurrence or recurrence of an adverse event. For the sake of clarity, the rested patient is still enrolled in the study until the physician investigator determines that the patient should not continue treatment, at which point the patient discontinues further treatment. In this context, duration of treatment refers to the time the patient is enrolled in the study, including all rest periods, until the treatment is interrupted.

Within the defined group, investigators may choose to reduce N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl a dose of either or both of acrylamide sulfonate and lenalidomide. In each case, the investigator may choose to gradually reduce the N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl group as follows. The dose of acrylamide benzene sulfonate. Patients in Group 1 were gradually reduced from 375 mg BID to 250 mg BID, and then gradually reduced from 250 mg BID to 125 mg BID N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)) Anilinopyrimidin-4-ylamino)phenyl)acrylamidobenzenesulfonate. Patients in group 1a can be gradually reduced from 375 mg BID to 250 mg BID, and then gradually reduced from 250 mg BID to 125 mg BID N -(3-(5-fluoro-2-(4-(2-methoxyethoxy)) Anilinopyrimidin-4-ylamino)phenyl)acrylamidobenzenesulfonate. Patients in Group 2 can be gradually reduced from 500 mg BID to 375 mg BID, and then gradually reduced from 375 mg BID to 250 mg BID N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)) Anilinopyrimidin-4-ylamino)phenyl)acrylamidobenzenesulfonate. Patients in group 2a can be gradually reduced from 500 mg BID to 375 mg BID, and then gradually reduced from 375 mg BID to 250 mg BID N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)) Anilinopyrimidin-4-ylamino)phenyl)acrylamidobenzenesulfonate. Patients in group 3a can be gradually reduced from 500 mg BID to 375 mg BID, and then gradually reduced from 375 mg BID to 250 mg BID N -(3-(5-fluoro-2-(4-(2-methoxyethoxy)) Anilinopyrimidin-4-ylamino)phenyl)acrylamidobenzenesulfonate. Patients in Group 3 can be gradually reduced from 500 mg BID to 375 mg BID, and then gradually reduced from 375 mg BID to 250 mg BID N -(3-(5-fluoro-2-(4-(2-methoxyethoxy)) Anilinopyrimidin-4-ylamino)phenyl)acrylamidobenzenesulfonate.

Investigators can choose to gradually reduce the dose of lenalidomide. As of August 6, 2013, the dose reduction steps are defined in Table 5:

In some cases, investigators may choose to gradually reduce the dose of lenalidomide according to Table 6:

It will be appreciated that dose reduction is not required and is provided only as a means of mitigating or alleviating one or more adverse events considered to be related to the study drug. Any such dose reduction does not alter patient registration; therefore any patient who has one or more dose reductions is still enrolled in the group to which they were originally assigned.

The patient registration dose should be based on the group that was opened at the time of registration. For the first cycle only, lenalidomide will be administered from day 1 to day 28 and N- (3-(5-fluoro-2-() will be administered on days 8-28 of the 28-day cycle. 4-(2-Methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl)propenylamine besylate. Thereafter, lenalidomide and N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidine will be administered according to Table 4 on a daily basis for a given 28-day period. 4-aminoamino)phenyl)propenylamine besylate.

If zero (0) in three (3) administered individuals suffers from DLT, the dose is considered tolerable. Dosing is allowed to be dosed by registration in the next higher dose only if no one (3) of the three (3) individuals before the registration in any group suffers from dose limiting toxicity (DLT). If one (1) of the first three (3) administered individuals in any group suffers from DLT during the first cycle, the other three (3) individuals should be enrolled in the dose group. When two (2) of the six (6) DLT evaluable individuals in the group are exposed to DLT, the dose should be considered NTD. The MTD is declared when at least six (6) individuals have registered and safely ended the first cycle at that dose. The MTD is defined as the last dose below the NTD, where zero (0) or one (1) DLT can assess that the individual is suffering from DLT during the first 28 day period.

During the dose escalation period, the decision to register the next higher dose group should be based on safety and review of the DLT evaluable patient. The OBE dose is defined as follows: 90 90% target occupancy rate in five (5) of six (6) individuals at any dose; and/or ̇ no additional exposure with dose increase; and/or ̇ Increased absolute lymphocyte count in four (4) of six (6) individuals during the first three 28-day cycles (lanes 1-3) 25%, not assessed as progressive disease (PD); and/or ̇ caused a reduction in total lymph node area in at least 50% of individuals at any dose or schedule 50%.

Research is still maintained for the individual until the individual is interrupted by the physician's decision due to disease progression, unacceptable toxicity, withdrawal consent, or any other reason. Preliminary evidence of efficacy will be assessed.

Results . "Complete response" (CR) according to IWCLL guidelines, 2008 (no LN > 1.5 cm, no hepatomegaly, splenomegaly, ALC < 4000 / μL, normal cell bone marrow <30% lymphocytes, ANC > 1500, platelets Counting >100,000 and Hgb > 11.0 g/dL) is defined. "Partial response" (PR) is via the IWCLL guidelines (at least 2 of the following guidelines: lymph node (LN) reduction) 50%; hepatomegaly is reduced 50%; splenomegaly is reduced 50%; ALC reduction 50%; and at least one of the following: platelet count >100,000; ANC > 1500 / μL or Hgb > 11.0 g / dL) was evaluated. "Node response" is defined as a reduction in total lymph node area of at least 50%. The PR status is an assessment of the laboratory values of the investigator based on physical examination of lymph nodes, spleen and liver and blood cell counts. Confirmed PR also includes imaging of tumor lesions by CT scan. Stable disease was determined by repeated CT scans based on unconfirmed CR clinical response.

Group 1. Three individuals were registered in Group 1 and N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl Combination treatment of acrylamide besylate sulfonate BID with lenalidomide QD. Of the three individuals initially enrolled in Group 1, one individual suffered from DLT (Level 3 Illusion). Because the study design does not allow for dose escalation (via registration at the next higher dose), unless only 1 of the 3 individuals or 6 individuals in the 3 doses suffered DLT during the first 28-day period at the current dose, The other three individuals were registered in group 1.

Figure 13 summarizes the disease status of patients in Group 1 up to October 16, 2013. Disease status was assessed according to the IWCLL guidelines for CLL diagnosis and treatment (Hallek, 2008). Among the six individuals enrolled in Group 1, Individual A exhibited progressive disease and discontinued treatment during the first cycle. In cycle 6, individual B exhibited a complete response confirmed by CT scan and bone marrow biopsy. MRD was confirmed to be negative by flow cytometry of bone marrow. Individuals C and F exhibited partial responses during the fourth cycle. Individuals D and E exhibited stable disease during the fourth cycle.

As of October 16, 2013, individual B was in cycle 10 and the two drugs were currently suspended due to AE diarrhea. Individual C was in cycle 8 and currently receives 375 mg of BID Compound 1 besylate. Individual D received 375 mg of BID Compound 1 benzenesulfonate throughout Cycle 5. Individual D withdrew to consent to the study. Individual E was in cycle 7 and currently receives group 1 dose (375 mg BID Compound 1 besylate and 10 mg QD lenalidomide). Individual F was in cycle 6 and received a 375 mg BID dose of Compound 1 besylate. Lenalidomide is suspended due to persistent neutropenia.

Group 2. Three individuals were registered in Group 2 and N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl Combination treatment of acrylamide besylate sulfonate BID with lenalidomide QD. Of the three individuals initially enrolled in Group 2, one subject suffered DLT (Grade 3 and Level 3 rash) during the first cycle and one individual was unable to assess DLT and needed to be replaced. The other four individuals were subsequently enrolled in Group 2 and treatment started.

Figure 14 summarizes the disease status of individuals in Group 2 up to October 16, 2013. Disease status was assessed according to the IWCLL guidelines for CLL diagnosis and treatment (Hallek, 2008). Among the seven individuals enrolled in group 2, reaction data of 4 individuals was obtained. Individuals A and B exhibited partial responses until the third cycle. Individuals C and D exhibited node responses until the second cycle. Individuals C and D have not met partial response criteria due to persistent lymphocytosis. Efficacy data for the remaining patients enrolled in Group 2 could not be obtained.

As of October 16, 2013, two individuals had been interrupted by disease progression during the fifth cycle. One individual was in cycle 4 and received 500 mg of BID Compound 1 besylate and a reduced dose of lenalidomide (5 mg QD) due to thrombocytopenia persisting at baseline. One individual was in cycle 3 and received a reduced dose of 375 mg of BID Compound 1 besylate; lenalidomide was suspended due to adverse event fatigue. One individual was in cycle 3 and received 500 mg of BID Compound 1 besylate; lenalidomide was suspended due to adverse event neutropenia. Another individual registered in Group 2 was interrupted by disease progression during the second cycle.

Group 2a. As of November 7, 2013, one patient was enrolled in Group 2a and is currently in the first cycle of treatment.

Expansion group . After DLT observations are completed in the dose escalation study, cumulative safety, PK, and PD data will be assessed to select initial RP2D. Preliminary RP2D or a more complete safety profile will be assessed in an expanded group of 24 individuals and further assessed for efficacy. If less than 9 of the 24 individuals are suffering from DLT, then this dose will be declared as RP2D to be used for further study. If greater than or equal to 9 of the 24 individuals are suffering from DLT, then this dose will be considered to have exceeded the MTD and the previous highest tolerated dose found in the dose escalation group of the study will be assessed in 24 individuals. The dose will continue to decrease in a gradual, progressive manner until less than 9 of the 24 individuals suffer from DLT.

In some cases, a physician investigator may choose to rest the patient during the study, during which time the patient does not receive treatment. For example, a physician investigator may choose to rest the patient due to the occurrence or recurrence of an adverse event. For the sake of clarity, the rested patient is still enrolled in the study until the physician investigator determines that the patient should not continue treatment, at which point the patient discontinues further treatment. In this context, duration of treatment refers to the time the patient is enrolled in the study, including all rest periods, until the treatment is interrupted.

Adverse events . For all groups, dose-limiting toxicity (DLT) was defined as the specified adverse event (AE) observed during the first 28-day period and considered to be treatment-related. Hematology DLT includes grade 4 anemia or thrombocytopenia according to CTCAE (version 4.03) or IWCLL guidelines, any of which causes a drop in blood threshold; grade 4 neutropenia over 5 days, even with granulocyte community stimulating factors (G-CSF) support; and grade 3 or higher febrile neutrophilia. Non-hematologic DLT includes non-hematologic AEs of grade 4 or higher at any duration within the first cycle; elevated levels of total bilirubin at grade 3, whether symptomatic or asymptomatic; and less than 24 after medical therapy Any grade 3 non-hematologic toxicity other than nausea, vomiting, and diarrhea; tumor lysis syndrome that has not progressed to grade 4 and is medically eliminated within 7 days; and transient, asymptomatic and rapidly reversible (in 7 days) Return to baseline or Level 1) Level 3 non-hematology laboratory abnormalities.

Individuals who have no disease progression at the end of the first 28-day treatment period are eligible to continue receiving N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidine-4- Amino)phenyl)propenylamine sulfonate and lenalidomide for a further 28-day period until (i) the patient suffers from unacceptable toxicity, (ii) potential malignancy progression, and (iii) patient withdrawal Agree, or (iv) the treating physician investigator otherwise determines that the patient should not continue treatment.

As of October 16, 2013, individuals in Group 1 reported hallucinations (1 patient; In the first cycle), group 1 was required; sepsis (1 patient; cycle 4); fever (1 patient; cycle 5) and neutropenia (1 patient; cycle 3).

As of October 16, 2013, individuals in Group 2 reported sepsis (1 patient; Cycle 2), thrombocytopenia (1 patient; 1st cycle, present at baseline), febrile neutrophil Reduction (1 patient; cycle 2) and neutropenia (1 patient; cycle 1). One individual reported fatigue and rash (Phase 1) and group 2 was needed.

Of the top 13 individuals (6 of group 1 and 7 of group 2) enrolled in groups 1 and 2, three reported mild tumor flare, which was before the end of the second week of treatment in all patients. eliminate.

Btk occupancy. N -(3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl)propenyl benzene sulfonate The covalent mechanism of action allows the development of covalent probes to detect free, uninhibited Btk in lysates derived from tissue culture, animal tissue or clinical samples. PBMC lysates were isolated from whole blood samples 30 minutes prior to dosing, 4 hours or 24 hours after dosing and incubated with biotinylated covalent probes. Uninhibited Btk was captured by covalent probes and quantified by ELISA. The %Btk occupancy was determined by calibration relative to the untreated control sample.

Btk target site occupancy ELISA: cell lysate or spleen homogenate with 1 μM N ¹ -(3-(3-(4-(3-acrylamidophenylamino)-5-methylpyrimidine-2) -aminoamino)phenoxy)propyl)-N ⁵ -(15-sided oxy-19-((3aS,4S,6aR)-2-yloxyhexahydro-1 H -thieno[3, 4-d]imidazol-4-yl)-4,7,10-trioxa-14-aza-n-pentyl)pentaneamine ( 2 ) in PBS, 0.05% Tween-20, 1% BSA solution together Incubate for 1 hour at room temperature. Compound 2 has the following structure:

Standards and samples were transferred to 96-well ELISA plates coated with streptavidin and mixed for 1 hour at room temperature on shaking. The α-Btk antibody (BD 611116, 1:1000 diluted in PBS + 0.05% Tween-20 + 0.5% BSA) was then incubated for 1 hour at room temperature. After washing, goat alpha-mouse-HRP (1:5000 diluted in PBS + 0.05% Tween-20 + 0.5% BSA) was added and incubated for 1 hour at room temperature. The ELISA was developed by the addition of tetramethylbenzidine (TMB) followed by the addition of a stop solution and read at OD 450 nm. A standard curve (11.7-3000 pg/μL) was generated using human full-length recombinant Btk protein and the mapping was fitted using a 4-parameter curve in the Gen5 software. Btk, which was undetected from the sample, was corrected to the total amount of μg of protein as determined by BCA protein analysis (Pierce Cat. No. 23225).

Example 2 Cell Titer Glo combination analysis

Analysis of N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl) in four model B cell carcinoma cell lines A combination of acrylamide sulfonate (compound 1 besylate) and lenalidomide to determine if the combination exhibits any synergistic effect.

DLBCL cell lines OCI-LY-10, WSU-DLCL2, Riva and TMD-8 were grown in RMPI + 10% FBS medium and plated in 96-well plates at 10,000 cells/well. Each cell strain was applied to a plurality of 96-well plates at 90 μl/well.

Based on the initial compound survival analysis of each cell line, the concentration range of the compound used for each cell strain was determined to achieve maximum and minimum cell survival rates in the case of uniform cell survival. A stock solution of 10 mM Compound 1 besylate in DMSO was prepared. The 10 mM stock solution was diluted between 3 x - 10 x to 3333 and 1000 nM and further diluted 3 fold in DMSO in a 10 point dilution series. The dilution series was diluted 50× in RMPI + 10% FBS medium and 5 μL of Compound 1 besylate was added to 95 μL of cell culture to yield a 1:1000 dilution.

Lenalidomide was prepared as a 10 mM stock solution in DMSO and diluted 1000 x in DMSO to the desired final concentration. The dilution series was diluted 200 x in RMPI + 10% FBS medium and 5 [mu]L of lenalidomide was added to 95 [mu]L of cell culture to yield a 1:1000 dilution. The final concentration of lenalidomide ranges from 41 to 10000 nM.

For combinatorial analysis, a concentration of lenalidomide to a one-part dilution series of compound 1 besylate was added. A combination of at least three and up to five different concentrations of lenalidomide and one of the three dilutions of Compound 1 besylate is analyzed. Compound 1 besylate and lenalidomide were applied to the cells on the second day after coating.

The number of viable cells was determined using CellTiter-Glo 72 hours after drug administration according to the manufacturer's instructions. Briefly, cells and CellTiter-Glo were allowed to equilibrate to room temperature. CellTiter-Glo was added to the cells at 1:1 (v/v) and analyzed. The results were read on a Molecular Devices spectramax L luminescence detector.

The luminescence readings were corrected to the percentage of the average control (DMSO treated) luminescence reading. The expected combination survival rate was calculated using the following formula: (Compound 1 benzenesulfonate % survival fraction) × (Lenalidomide % survival fraction) / 100

The survival rate curve was drawn using Microsoft Excel with the XLfit plugin. A synergistic ("volcano") map is generated by plotting the difference between the observed and expected combination of treatment values. The results are presented in Figures 1-12 . The figures depicted in the drawings are illustrative unless otherwise indicated.

The dose response curves in Figures 1, 3, 5, 7, 9 and 11 depict the combination of Compound 1 besylate and lenalidomide in the cell line OCI-LY-10 (Figure 1), WSU-DLCL2 (Figure 3 ), Riva (Figures 5, 7 and 9) and TMD-8 (Figure 11). The response of the cells to the concentration of Compound 1 besylate is indicated by ●. The response of the cells to the test concentration of lenalidomide is indicated by □. The expected (or calculated) activity of the combination is indicated by Δ. The observed activity of this combination is indicated by ◇. When the expected or calculated activity of the combination is equal to the observed activity (i.e., when the lines represented by Δ and ◇ overlap), the combination is additive.

Figure 1 presents dose response curves observed with compound 1 besylate and a specific concentration of lenalidomide (3000 nM) in OCI-LY-10 cell lines. The results presented in Figure 1 appear to show an additive effect and are confirmed by additional experiments (data not shown). Figure 1 can thus be considered to represent data obtained in the OCI-LY-10 cell line for the combination of Compound 1 besylate and lenalidomide. Figure 2 presents a "volcano" plot of Compound 1 besylate and lenalidomide in the OCI-LY-10 cell line. The volcano map shown in Figure 2 is generated using data from experiments conducted at different concentrations of lenalidomide and represents an additional experiment in the OCI-LY-10 cell line. Confirm the results by additional experiments (data not shown). Compound 1 besylate and lenalidomide therefore appeared to be additive in the OCI-LY-10 cell line.

Figure 3 depicts a specific dose response curve for Compound 1 besylate and a specific concentration of lenalidomide (3333 nM) in the WSU-DLCL2 cell line. The results depicted in Figure 3 were confirmed by additional experiments (data not shown). Figure 3 thus can be considered to represent data obtained in the WSU-DLCL2 cell line for the combination of Compound 1 besylate and lenalidomide. The results presented in Figure 3 appear to show an additive effect. Figure 4 presents a "volcano" plot of Compound 1 besylate and lenalidomide in a WSU-DLCL2 cell line. The combination of lenalidomide with 37 nM, 111 nM, 370 nM and 1111 nM of Compound 1 besylate was also tested. In this experiment, there was no significant synergy at these concentrations (data not shown). Compound 1 besylate and lenalidomide therefore appeared to be additive in the WSU-DLCL2 cell line.

Figure 5 depicts a specific dose response curve for Compound 1 besylate and lenalidomide (3333 nM) in Riva cell lines. At this concentration, Compound 1 besylate and lenalidomide appeared to be synergistic (indicated by arrows). Figure 6 presents a "volcano" plot of Compound 1 besylate and lenalidomide in Riva cell lines at concentrations of lenalidomide at 41 nM, 1111 nM and 3333 nM. At certain concentrations, Compound 1 besylate and lenalidomide appeared to be synergistic (indicated by arrows).

Figure 7 presents a specific dose response curve for Compound 1 besylate and lenalidomide (333 nM) in the Riva cell line. Compound 1 besylate and lenalidomide appear synergistic in this figure (indicated by arrows). Figure 8 presents a "volcano" plot of a specific experiment using Compound 1 besylate and lenalidomide in Riva cell lines at lenalidomide concentrations of 41 nM, 123 nM and 370 nM. Compound 1 and benzenesulfonate lenalidomide appears to be synergistic (indicated by arrows) in this figure.

Figure 9 presents another dose response curve for Compound 1 besylate and lenalidomide (333 nM) in the Riva cell line. Compound 1 besylate and lenalidomide appear to be additive in this figure. Figure 10 presents a "volcano" plot of another experiment of Compound 1 besylate and lenalidomide in Riva cell lines at concentrations of lenalidomide at 41 nM, 123 nM and 370 nM. Compound 1 besylate and lenalidomide appeared to be additive in Figure 10 . The data in Figures 7-10 appear to show that the combination of Compound 1 besylate and lenalidomide is not antagonistic and may be synergistic.

Figure 11 presents a specific dose response curve for Compound 1 besylate and lenalidomide (3000 nM) in the TMD-8 cell line. Compound 1 besylate and lenalidomide appeared to be synergistic (indicated by arrows). Figure 12 presents a "volcano" plot of a specific experiment using Compound 1 besylate and lenalidomide in a TMD-8 cell line at concentrations of lenalidomide at 300 nM, 1000 nM, and 3000 nM. Compound 1 besylate and lenalidomide appear synergistic in this figure (indicated by arrows). The combination of lenalidomide with Compound 1 besylate at concentrations of 100 nM, 300 nM and 1000 nM was also tested and significant synergy was confirmed (data not shown).

Compound 1 besylate and lenalidomide appeared to be additive in OCI-LY-10 ( Figures 1 and 2 ) and WSU-DLCL2 ( Figures 3 and 4 ) cell lines.

In Figures 5 and 6 , Compound 1 besylate and lenalidomide appeared to be synergistic in the Riva cell line at 3333 nM concentration of lenalidomide. Compound 1 besylate and lenalidomide appeared synergistic in Figures 5 and 6 (indicated by arrows). In Figures 7 and 8 , Compound 1 besylate and lenalidomide exhibited some significant synergy at 333 nM concentration of lenalidomide. In Figures 9 and 10 , Compound 1 besylate and lenalidomide appeared to be additive at 333 nM concentration of lenalidomide. In Fig. 11 and Fig. 12 , it was confirmed that the compound 1 besylate and lenalidomide were synergistic in the TMD-8 cell line. Thus, Compound 1 besylate and lenalidomide appear to exhibit a synergistic effect in at least the TMD-8 cell line. It also appears that some evidence of synergy has been observed in Riva cell lines.

Example 3

Screen for a specific irreversible BTK inhibitor Compound 2 against 342 kinases to determine kinase activity and/or selectivity:

The binding assay system for understanding the activity profile of the kinase was based on the HotSpot technique (Reaction Biology Corp.; Malvern, PA, USA) and was filtered using a radioisotope based P81. Compound 2 was dissolved in pure DMSO to make a 10 mM stock solution and serial dilutions were made to a final 3 [mu]M test concentration. The substrate for each of the kinases tested for Compound 2 was prepared daily in reaction buffer (substance information is available on the Reaction Biology Corp. website). Any desired cofactor is then added to the substrate solution. Identification and selection of appropriate cofactors for various kinases is within the skill of those skilled in the art. See, for example, Handbook of Assay Development in Drug Discovery , Lisa K. ed. Minor, 2006: CRC Press, Boca Raton, Florida; Gao et al., "A broad activity screen in support of a chemogenomic map for kinase signalling research and drug discovery" , Biochem J. 2013, 451(2): 313-28; and Eglen et al., "Drug discovery and the human kinome: Recent trends", Pharmacology & Therapeutics 2011, 130(2): 144-156. The kinase is then added to the substrate and gently mixed. Compound 2 (5 nL) was then added to the kinase reaction mixture by sonic droplet ejection and pre-incubated for 30 minutes at room temperature. ³³ P-ATP (100 μM) was delivered to the reaction mixture to initiate the reaction. Thereafter, it was incubated at room temperature for 2 hours. The reaction was terminated and any unreacted phosphate was washed away with 0.1% phosphoric acid and subsequently detected using proprietary techniques. Studies were performed in duplicate and staurosporine (a non-selective, ATP-competitive kinase inhibitor) was used as a 10-dose IC50 model with 3 fold serial dilutions starting at 1 μM, 50 μM or 100 μM. A positive control in the middle. DMSO was used as a negative control.

Determination of percentage of inhibition. Percent inhibition of kinase by test compound (eg Compound 2 ) is determined according to the following formula: percent inhibition = [(kinase activity of negative control) - (kinase activity in the presence of, for example, test compound of compound 2 ) / ( Kinase activity of the negative control)] × 100. Percent inhibition is expressed as the average value in the case of more than one (eg, duplicate) analysis.

Table 7 illustrates the average percent inhibition of Compound 2 against various kinases:

Example 4 Dose escalation study of rituximab

N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilinyl)pyrimidin-4-ylamino)phenyl)propenyl benzene sulfonic acid is used in this study. The capsule of salt corresponds to Table 1 in Example 1.

The lenalidomide capsules correspond to the lenalidomide capsules listed in Tables 2 and 3 of Example 1.

Rituximab is provided to the physician/investigator in a 10 mg/mL vial containing 100 mg/10 mL or 500 mg/50 mL. Rituximab was diluted to a dose of 1 mg/mL, 2 mg/mL, 3 mg/mL or 4 mg/mL with 5% dextrose in water or 0.9% sodium chloride prior to administration. Thereafter, rituximab was administered as an infusion as described in Table 8, below, at a dose of 1 mg/mL to 4 mg/mL.

Research design

Individuals with recurrent or refractory CLL or SLL with at least one prior treatment regimen were enrolled in the "3+3" dose escalation and expansion study to determine the resistance of Compound 1 , lenalidomide and rituximab Subjected dose (NTD), optimal biological effect dose (OBE) and maximum tolerated dose (MTD). Approximately 30-42 patients are expected to be enrolled in the study.

Study treatment is administered as scheduled at the prescribed dose over a 28 day period until disease progression, toxicity is unacceptable, or is interrupted for any other reason. The individual will continue with the starting dose until a preliminary recommended phase 2 dose (RP2D) is determined, at which point it can be converted to a preliminary RP2D.

N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilinyl)pyrimidin-4-ylamino)phenyl)propene will be administered according to the groups listed in Table 8. Indole benzene sulfonate (compound 1 besylate), lenalidomide and rituximab:

Within each group, individuals will be treated according to the group in Table 8 and assessed for safety, tolerability, and DLT, as well as pharmacokinetics ("PK"), pharmacodynamics ("PD"), and disease response.

(PO) Compound 1 besylate and lenalidomide were administered according to Table 8.

Rituximab is administered as a single intravenous (IV) infusion. The initial infusion in the first cycle will be administered at 375 mg/m ² ; the subsequent infusion in the second to sixth cycles will be administered at 500 mg/m ² . The administration of rituximab will occur on the first day of each 28-day cycle. After the infusion of the sixth cycle, rituximab will be discontinued. If appropriate, the individual can continue to be treated with Compound 1 besylate and/or lenalidomide. The first infusion of rituximab will be at a rate of 50 mg/h. In the absence of infusion toxicity, the infusion rate will increase by 50 mg/h every 30 minutes, reaching a maximum of 400 mg/h. Each subsequent infusion will start at 100 mg/h. In the absence of infusion toxicity, the infusion rate will increase by 100 mg/h in 30 minute intervals to a maximum of 400 mg/h.

If lenalidomide does not achieve tolerance and/or desired response after 2 cycles at 5 mg (i.e., after a total of at least 3 28-day cycles have ended), the dose of lenalidomide can be increased to 10 mg QD.

Claims (28)

A method of treating, stabilizing or ameliorating the severity or progression of one or more BTK-related diseases and conditions, comprising administering Compound 1 ( N- (3-(5-fluoro-2-(4)) to a patient in need thereof) -(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl)propenylamine): Or a pharmaceutically acceptable salt thereof, and lenalidomide. The method of claim 1, wherein the disease or condition associated with BTK is chronic lymphocytic leukemia. The method of claim 1, wherein the disease or condition associated with BTK is a small lymphocytic lymphoma. The method of claim 2 or 3, wherein Compound 1 is administered twice a day. The method of claim 4, wherein the compound 1 is in the form of a besylate salt. The method of claim 5, wherein the lenalidomide is administered once a day. The method of claim 6, wherein the compound 1 and lenalidomide are each administered in an oral dosage form. The method of claim 7, wherein Compound 1 and lenalidomide are each administered for at least one 28 day period. The method of claim 2 or 3, wherein the patient has failed at least one prior therapy. The method of claim 5, wherein the compound 1 comprises from about 10% to about 50% of N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidine-4 The composition of the composition of the group -aminoamino)phenyl)propenylamine besylate is administered. The method of claim 10, wherein the composition comprises about 42% N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino) Phenyl) acrylamide benzene sulfonate. The method of claim 1, additionally comprising administering an anti-CD20 antibody. A method of preventing, treating, stabilizing or ameliorating the severity or progression of a disease or disorder selected from the group consisting of chronic lymphocytic leukemia and small lymphocytic lymphoma, the method comprising administering a therapeutically effective amount to a patient in need thereof Compound 1 or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of lenalidomide, wherein the therapeutically effective amount of Compound 1 is from about 750 mg to about 1000 mg per day. The method of claim 13, wherein the therapeutically effective amount of Compound 1 is about 375 mg BID. The method of claim 13, wherein the therapeutically effective amount of Compound 1 is about 500 mg BID. The method of claim 14 or 15, wherein the therapeutically effective amount of lenalidomide is about 10 mg. The method of claim 14 or 15, wherein the therapeutically effective amount of lenalidomide is about 5 mg. The method of claim 16 or 17, wherein the lenalidomide is administered once a day (QD). The method of claim 13, additionally comprising administering an anti-CD20 antibody. A system for treating, stabilizing or ameliorating the severity of one or more diseases or conditions associated with BTK, the system comprising Compound 1, or a pharmaceutically acceptable salt thereof, and lenalidomide. The method of claim 20, wherein the system further comprises an anti-CD20 antibody. A method of treating, stabilizing or ameliorating the severity or progression of one or more BTK-related diseases and conditions comprising administering to a patient in need thereof an irreversible BTK inhibitor and lenalidomide, wherein the irreversible BTK inhibitor has a kinase selected from the following Not more than about 50% inhibition: c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, BRAF, RIPK3, ARAF and SRMS, or a combination thereof . The method of claim 22, wherein the patient anti-CD20 antibody is additionally administered. The method of claim 22, wherein the irreversible BTK inhibitor has no more than about 30% inhibition of a kinase selected from the group consisting of c-Kit, RIPK2, HCK, EPHA6, LYN, CSK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, BRAF, RIPK3, ARAF and SRMS, or a combination thereof. The method of claim 22, wherein the irreversible BTK inhibitor has no more than about 10% inhibition of a kinase selected from the group consisting of: EPHA6, LYN B, FRK/PTK5, BRAF, RIPK3, ARAF, and SRMS, or a combination thereof. The method of claim 22, wherein the irreversible BTK inhibitor has a percent inhibition of LYN of no more than about 20-30%. The method of claim 22, additionally comprising administering an anti-CD20 antibody. A method of treating, stabilizing or ameliorating the severity or progression of one or more BTK-related diseases and conditions, comprising administering Compound 1 ( N- (3-(5-fluoro-2-(4)) to a patient in need thereof) -(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl)propenylamine): Or a pharmaceutically acceptable salt thereof, lenalidomide and an anti-CD20 antibody.