TW201427667A - 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

The present application claims priority to US Provisional Application No. 61/728,701, filed on Nov. 20, 2012, and No. 61/870,720, filed on August 27, 2013, each of which The manner of full reference is incorporated herein.

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. Research Studies 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, Clinical anti-tumor response in NHL) and 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-associated diseases and conditions, comprising administering Compound 1 to rituximab to a patient in need thereof A combination of (rituximab).

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 compositions of rituximab.

In some embodiments, the methods provided comprise administering a combination of Compound 1 and rituximab to a patient in need thereof, wherein Compound 1 is administered once a day. In some embodiments, the method comprises providing to a patient in need of the compound 1 in combination with rituximab of administration, wherein the compound is administered twice a day. In some of these embodiments, rituximab is administered once in a 28 day period. Thus, in some embodiments, the methods provided comprise administering a combination of Compound 1 and rituximab to a patient in need thereof, wherein Compound 1 is administered twice a day and rituximab is within 28 days of the cycle. Vote once.

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

In some embodiments, the methods provided comprise administering Compound 1 , rituximab, fludarabine, and cyclophosphamide to a patient in need thereof.

In some embodiments, the methods provided comprise administering Compound 1 , rituximab, and bendamustine to a patient in need thereof.

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 present invention provides a method of treating, stabilizing or reducing the severity or progression of chronic lymphocytic leukemia (CLL), the method comprising the administration to a patient in need thereof with a compound of rituximab combination.

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

In some embodiments, the provided therapy comprises orally administering to a patient a combination of Compound 1 and rituximab. In some embodiments, Compound 1 and rituximab to each administered as a pharmaceutical formulation. In some embodiments, a pharmaceutical formulation comprising Compound 1 is a capsule formulation. In some embodiments, the pharmaceutical formulation comprising rituximab is an intravenous (IV) formulation.

In some embodiments, the invention also provides a dosing regimen and schedule for administering a combination of Compound 1 and rituximab to a patient in need thereof. The methods, dosing regimens and plans for administration of the combination are described in further detail below.

Figure 1 presents a response assessment of patients enrolled in groups 1 and 2 up to October 16, 2013.

definition

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 particular 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 despite clinical There are minor differences in inactive components, but biological products are highly similar to reference products; (b) animal studies (including toxicity assessment); and/or (c) clinical studies (including immunogenicity and pharmacokinetics or pharmacodynamics) Evaluation), which is sufficient to demonstrate the safety, purity and efficacy (eg, in terms of product safety, purity and efficacy) in the biological product of the approved and intended use and approval of the reference product (eg, in terms of product safety, purity and efficacy) There is no clinically meaningful difference between the reference product and the reference product).

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.

As used herein, the terms "combination,""in combination with," or "combination therapy" refer to the condition in which two or more different pharmaceutical agents are administered in an overlapping regimen such that the individual is simultaneously exposed to both agents. In some embodiments, the combinations are directed to a separate dosage form of Compound 1 and rituximab administered to the individual. In some embodiments, the combinations are directed to an individual to administer a separate dosage form of Compound 1 and rituximab, wherein Compound 1 is administered prior to, during, or after administration of rituximab. In some embodiments, simultaneous or simultaneous exposure of Compound 1 and rituximab is achieved via different dosing regimens appropriate for each therapeutic agent. For example, Compound 1 can be administered once or twice daily for one or more 28-day cycles, while rituximab can be administered once over a 28-day period.

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 2, 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 comparisons between them so that they can be based on observations The difference or similarity is 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 a reference kinase inhibitor has a 50% inhibition of the kinase relative to a control, if the other inhibitor has about 40% to about 60% inhibition of the same kinase relative to the control, then the other inhibitor should be considered Comparable 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 rituximab.

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 a time dependence, whereby the degree of inhibition increases as the inhibitor is contacted 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.

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 and rituximab to a patient in need thereof Monoclonal antibody. In some of these embodiments, the methods provided additionally comprise administering fludarabine and cyclophosphamide.

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 and rituximab to a patient in need thereof Monoclonal antibody and bendamustine.

In some embodiments, the irreversible BTK inhibitor is covalently bound to Cys 481 of BTK.

In some embodiments, the irreversible BTK inhibitor has activity against one or more kinases selected from the kinases listed in Table 3 below.

In some embodiments, the irreversible a BTK inhibitor selected from Table 3 for the kinase or kinase inhibitor composition can have a reference percentage of inhibition comparable. 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 2.

In some embodiments, the irreversible BTK inhibitor has a percent inhibition comparable to Compound 2 for one or more kinases selected from Table 3 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 3 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 3 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 3 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 2.

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 2.

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 2.

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 2.

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 2.

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 2.

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 In combination with rituximab, 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 2.

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 2.

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 2.

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 2.

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 2.

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 2.

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 a combination of an irreversible BTK inhibitor and rituximab, wherein the irreversible The BTK inhibitor has 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 2.

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 2.

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 2.

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 2.

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 2.

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 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 2.

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 2.

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 2.

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 2.

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 2.

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 2.

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 2.

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 2.

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 inhibits Btk activity by binding to the Btk adenosine triphosphate (ATP) binding site with high affinity and covalently binding to the Btk protein forming target, providing rapid, complete and prolonged inhibition of Btk activity in vitro and in vivo.

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).

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 combination of a BTK inhibitor (ie, Compound 1 ) 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 a patient in need thereof a combination of Compound 1 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 affinities determinant 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 5nM, 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. Thus, in some embodiments, the methods provided comprise administering Compound 1 and 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 and 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 embodiments, 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 in a nutrient medium containing the antibiotic gentamicin The suspension culture of mammalian cells (Chinese hamster ovary) is produced. 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 in combination with fludarabine and cyclophosphamide, and in combination with methotrexate for rheumatoid Patients with arthritis. Rituximab is also approved for the treatment of non-Hodgkin's lymphoma, Wegener's Granulomatosis and Microscopic Polyangiitis.

In some embodiments, the methods provided comprise administering a combination of Compound 1 and rituximab to a patient in need thereof, wherein the patient is additionally treated with fludarabine and cyclophosphamide according to the approved indications .

I. General dosing plan

As described herein, the methods provided comprise administering Compound 1 and an anti-CD20 antibody (eg, rituximab, orfarizumab, opiconumab, etc.) to a patient in need thereof. These methods additionally comprise administration of (i) fludarabine and cyclophosphamide or (ii) bendamustine, as appropriate. It will be understood that each therapeutic agent (ie, Compound 1 , anti-CD20 antibody, fludarabine, cyclophosphamide, and bendamustine) may be synchronized or sequentially as part of a dosing regimen (eg, Compound 1 may be resistant Administration of CD20 antibody and/or (i) fludarabine and cyclophosphamide or (ii) bendamustine before, during or after administration, and vice versa). For example, Compound 1 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 some embodiments, Compound 1 and the anti-CD20 antibody can be one or more hours, one or more days or one prior to administration of (i) fludarabine and cyclophosphamide or (ii) bendamustine Or vote for more than a few weeks.

In some embodiments, the invention provides for treating, stabilizing or reducing one or more A method of severity or progression of a disease or condition 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 rituximab to a patient in need thereof. In some such embodiments, Compound 1 and rituximab are each administered as a composition additionally comprising one or more pharmaceutically acceptable excipients. In some embodiments, a method provided comprises administering each of Compound 1 , rituximab, fludarabine, and cyclophosphamide. In some such embodiments, Compound 1 , rituximab, fludarabine, and cyclophosphamide are each administered as a composition additionally comprising one or more pharmaceutically acceptable excipients. In some embodiments, the methods provided comprise administering each of Compound 1 , Rituximab, and bendamustine. In some such embodiments, Compound 1 , rituximab, and bendamustine are each administered as a composition additionally comprising one or more pharmaceutically acceptable excipients.

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 rituximab. 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 rituximab. In some embodiments, a method provided comprises administering to a patient in need thereof a therapeutically effective amount of each of Compound 1 , Rituximab, Fludarabine, and Cyclophosphamide. In some embodiments, the methods provided comprise administering to a patient in need thereof a therapeutically effective amount of each of Compound 1 , Rituximab, and bendamustine.

In some embodiments, the methods provided comprise administering a combination of Compound 1 and rituximab, wherein Compound 1 is administered once daily ("QD"). In some embodiments, the methods provided comprise administering a combination of Compound 1 and rituximab, wherein Compound 1 is administered twice daily ("BID"). 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 each of Compound 1 , rituximab, fludarabine, and cyclophosphamide, wherein Compound 1 is administered twice daily ("BID" ). In some embodiments, the methods provided comprise administering each of Compound 1 , rituximab, and bendamustine, wherein Compound 1 is administered twice daily ("BID").

In some embodiments, the methods provided comprise administering a combination of Compound 1 and rituximab, wherein rituximab is administered once in a 28 day period. In some embodiments, the methods provided comprise administering a combination of Compound 1 and rituximab, wherein rituximab is administered on Day 1 or Day 2 of the first cycle. In some embodiments, the methods provided comprise administering a combination of Compound 1 and rituximab, wherein rituximab is administered on Day 1 of a 28 day cycle. In some of these embodiments, rituximab is administered on the first day of cycle 2-6. In some embodiments, rituximab is administered on the first day of cycle 2-5. In some embodiments, rituximab is administered on the first day of cycles 2-4. In some embodiments, rituximab is administered on the first day of the 2-3th cycle. In some embodiments, the methods provided comprise administering each of Compound 1 , rituximab, fludarabine, and cyclophosphamide, wherein rituximab is administered over a 28 day period once. In some of these embodiments, rituximab is administered on the first or second day of the 28 day cycle.

In some embodiments, the methods provided comprise administering each of Compound 1 , rituximab, and bendamustine, wherein rituximab is administered once in a 28 day period. In some of these embodiments, rituximab is administered on the first or second day of the 28 day cycle.

In some embodiments, the methods provided comprise administering a combination of Compound 1 and rituximab, wherein Compound 1 is administered twice daily and rituximab is administered once over a 28 day period. In some embodiments, the methods provided comprise administering a combination of Compound 1 and rituximab, wherein Compound 1 is administered twice daily and rituximab is on Day 1 or Day 2 of Cycle 1. Cast. In some embodiments, the methods provided comprise administering a combination of Compound 1 and rituximab, wherein Compound 1 is administered twice daily and rituximab is administered on Day 1 of a 28 day cycle. In some of these embodiments, rituximab is administered on the first day of cycle 2-6. In some embodiments, the methods provided comprise administering each of Compound 1 , rituximab, fludarabine, and cyclophosphamide, wherein Compound 1 is administered twice daily and rituximab The monoclonal antibody was administered once in a 28-day cycle. In some of these embodiments, rituximab is administered on the first or second day of the 28 day cycle.

In some embodiments, the methods provided comprise administering each of Compound 1 , Rituximab, and bendamustine, wherein Compound 1 is administered twice daily and rituximab is at 28 Vote once in the day cycle. In some of these embodiments, rituximab is administered on the first or second day of the 28 day cycle.

In some embodiments, Compound 1 and rituximab are each administered as a pharmaceutically acceptable composition. In some embodiments, a pharmaceutically acceptable composition comprising Compound 1 is formulated into an oral dosage form. In some embodiments, the oral dosage forms are capsules. In some embodiments, a pharmaceutically acceptable composition comprising rituximab is formulated into an intravenous composition. In some embodiments, fludarabine, cyclophosphamide, and bendamustine are 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 .

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.

Fludara® is commercially available as a vial containing 50 mg of fludarabine phosphate, 50 mg of mannitol, and a sterile lyophilized solid cake conditioned to adjust the pH to 7.7. The pH of the final solution ranged from 7.2 to 8.2. The solid cake was reconstituted with 2 mL of sterile water for injection USP to produce a solution containing 25 mg/mL fludarabine phosphate intended for intravenous administration.

Cyclophosphamide (Cytoxan®) is commercially available as a sterile powder which can be prepared for parenteral administration by infusion by, for example, reconstitution in 0.9% sterile sodium chloride (5 mL per 100 mg of anhydrous powder). Alternative solutions for reconstitution can be found, for example, in the package insert.

Benzamustine (Treanda®) is commercially available as a single use vial containing 100 mg of bendamustine hydrochloride as a lyophilized powder. The powder was reconstituted to a final concentration of 5 mg/mL with 20 mL of sterile water for injection USP. Immediately prior to the infusion, the 5 mg/mL reconstituted solution was transferred to a 500 mL infusion bag containing 0.9% sodium chloride injection USP. Or, you can put 5 The mg/mL reconstituted solution was transferred to a 500 mL infusion bag containing 2.5% dextrose/0.45% sodium chloride injection USP. The final concentration of bendamustine hydrochloride in the infusion bag should be about 0.2-0.6 mg/mL.

In some embodiments, the methods provided comprise administering a combination of Compound 1 and rituximab 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 refer to a patient in need of the provided treatment regimen for 28 consecutive days. In some embodiments, the combination of Compound 1 and rituximab 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 rituximab is administered 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 rituximab is administered at least thirteen, at least fourteen, at least fifteen, at least sixteen, at least seventeen, at least eighteen, at least ten Nine or at least twenty 28-day cycles. In some embodiments, a combination of Compound 1 and rituximab is administered to a patient for a lifetime.

In some embodiments, the methods provided comprise administering to a patient in need thereof, each of Compound 1 , rituximab, fludarabine, and cyclophosphamide, wherein rituximab is administered At least one, two, at least three, at least four, at least five, or at least six 28-day cycles of each of fludarabine and cyclophosphamide.

In some embodiments, the combination of Compound 1 and rituximab is administered for at least six 28 day cycles, and Compound 1 is administered again for at least one 28 day period. In some embodiments, the combination of Compound 1 and rituximab is administered for at least six 28-day cycles, and Compound 1, two, three, four, five, six, seven, eight are re-administered. 9, nine, ten, eleven, twelve, thirteen or fourteen 28-day cycles. In some embodiments, the combination of Compound 1 and rituximab is administered for at least six 28 day cycles, and Compound 1 is administered for a lifetime of the patient. 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 administered on Day 1 of the 28-day cycle. Tubazumab. In some embodiments, Compound 1 is administered from Day 1 to Day 28 of one or more 28-day cycles and rituximab is administered on Day 1 or Day 2 of the 28-day cycle.

In some embodiments, two adjacent 28 day periods may be separated by a rest period. Such rest periods may be one, two, three, four, five, six, seven or seven days, during which no compound 1 or rituximab 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 rituximab, wherein the patient has failed at least one prior therapy. In some embodiments, the methods provided comprise administering Compound 1 , rituximab, fludarabine, and cyclophosphamide to a patient in need thereof, wherein the patient has failed at least one prior therapy. In some embodiments, the methods provided comprise administering Compound 1 , rituximab, and bendamustine to a patient in need thereof, 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 compound is a therapeutically effective amount of 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 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 from about 625 mg to about 875 mg, or about 625 mg to 750mg, or from about 750mg to about 1250mg, or from about 750mg to about 1125mg, or from about 750mg to about 1000mg, or from about 875mg to about 1250mg, or from about 875mg to about 1125mg, or from about 875mg to about 1000mg.

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 pharmaceutical composition comprising a unit dose of Compound 1 in combination with rituximab. 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, a method is provided comprising administering to a patient in need thereof a pharmaceutical composition comprising rituximab, wherein rituximab is administered by 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 great influence on anti-IgM-induced B cell proliferation and inhibition of non-thymus-dependent type II antibodies The original immune response (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 rituximab 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.

Btk cells are limited in expression and are mainly limited to B lymphocytes, monocytes and hypertrophy Cytoplasmic or basophilic white blood cells. 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. Recently, it has been reported that one of the CLLs retains in vitro BCR signaling, and immunoglobulin heavy chain gene somatic mutation (IgVH) is an important determinant of the 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 Waldenstrom's macroglobulinemia (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 rituximab

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. The most convincing is that 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 Objective anti-tumor response and good tolerance in patients with lymphoma (FL), 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 for the treatment of 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.

Rituximab has also been shown to exhibit good activity against patients with relapsed/refractory CLL. In one study, the combination of rituximab and fludarabine/cyclophosphamide was evaluated in 408 CLL patients and showed a 86% response rate with respect to fludarabine/cyclophosphamide alone. Compared to the observed 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 rituximab for the treatment of CLL and SLL. Compound 1 can be found to be effective in CLL patients, whether in a single agent or combination, including but not limited to patients who have exhibited 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 provides methods of treating, stabilizing or ameliorating the severity or progression of one or more BTK-related diseases and conditions, comprising administering Compound 1 to rituximab to a patient in need thereof combination.

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 was administered a combination of Compound 1 and rituximab. In some embodiments, a method provided comprises administering to a patient in need thereof, each of Compound 1 , rituximab, fludarabine, and cyclophosphamide. In some embodiments, the methods provided comprise administering to each of the patients in need, Compound 1 , Rituximab, and Bendamustine.

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 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, a method provided comprises administering to a patient in need thereof a composition comprising each of Compound 1 , Rituximab, Fludarabine, and Cyclophosphamide. In some embodiments, a method provided comprises administering to a patient in need thereof a composition comprising each of Compound 1 , Rituximab, and Bendamustine.

In some embodiments, the composition comprising rituximab additionally comprises one or more pharmaceutically acceptable excipients. In some such embodiments, the composition comprising rituximab is formulated into an intravenous dosage form.

In some embodiments, the methods have provided comprising administering to the patient a unit dose of the compound and the needs of a unit dose of rituximab combination of rituximab. 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, a method is provided comprising administering to a patient in need thereof a pharmaceutical composition comprising rituximab, wherein rituximab is administered by 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 rituximab, wherein the patient has failed at least one prior therapy. In some embodiments, the methods provided comprise administering Compound 1 , rituximab, fludarabine, and bendamustine to a patient in need thereof, wherein the patient has failed at least one prior therapy. In some embodiments, the methods provided comprise administering Compound 1 , rituximab, and bendamustine to a patient in need thereof, 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 a combination of from about 500 mg to about 1250 mg of Compound 1 and from about 375 mg/m ² to about 500 mg/m ² rituximab. In some embodiments, the methods provided comprise administering to a patient in need thereof a combination of from about 750 mg to about 1000 mg of Compound 1 and from about 375 mg/m ² to about 500 mg/m ² rituximab. In some embodiments, the method comprises providing to a patient in need of administration and have from about 500mg to about 1250mg of Compound 1, about 375mg / m ² to about 500mg / m ² rituximab, from about 25mg / m ² of fluoro Rabin and about 250 mg/m ² cyclophosphamide. In some embodiments, the methods provided comprise administering from about 750 mg to about 1000 mg of Compound 1 , about 375 mg/m ² to about 500 mg/m ² of rituximab, and about 70 mg/m ^{2 of} benzoic acid to a patient in need thereof. Mustin.

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 375 mg BID to about 500 mg BID Compound 1 and about 375 mg/m ² to about 500 mg/m ² rituximab. In some of these embodiments, rituximab was administered once in a 28 day period. In some embodiments, the methods provided comprise administering from about 250 mg to about 500 mg of BID Compound 1 , about 375 mg/m ² to about 500 mg/m ² of rituximab, about 25 mg/m ^{2 of} fluoride to a patient in need thereof. Dalazine and about 250 mg/m ² cyclophosphamide, wherein rituximab was administered on the first day of the 28-day cycle. In some of these embodiments, each of fludarabine and cyclophosphamide is administered on days 1-3 of the 28 day cycle. In some embodiments, the methods provided comprise administering from about 750 mg to about 1000 mg of Compound 1 , about 375 mg/m ² to about 500 mg/m ² of rituximab, and about 70 mg/m ^{2 of} benzoic acid to a patient in need thereof. Mestin, in which rituximab was administered on the first day of the 28-day cycle. In some of these embodiments, bendamustine was administered on days 1 and 2 of the 28 day cycle.

In some embodiments, the methods provided comprise administering about 125 mg of BID Compound 1 and about 375 mg/m ² of rituximab to a patient in need thereof. In some of these embodiments, the methods provided additionally comprise administering about 25 mg/m ² fludarabine and about 250 mg/m ² cyclophosphamide. In some embodiments, the methods provided comprise administering to a patient in need thereof about 125 mg BID Compound 1 , about 375 mg/m ² rituximab, and about 70 mg/m ² bendamustine.

In some embodiments, the methods provided comprise administering about 125 mg of BID Compound 1 and about 500 mg/m ² of rituximab to a patient in need thereof. In some such embodiments, the method provided further comprises the administration of about 25mg / m ² fludarabine and about 250mg / m ² cyclophosphamide. In some embodiments, the methods provided comprise administering to a patient in need thereof about 125 mg BID Compound 1 , about 500 mg/m ² rituximab, and about 70 mg/m ² bendamustine.

In some embodiments, the methods provided comprise administering about 250 mg of BID Compound 1 and about 375 mg/m ² of rituximab to a patient in need thereof. In some of these embodiments, the methods provided additionally comprise administering about 25 mg/m ^{2 of} fludarabine and about 250 mg/m ^{2 of} cyclophosphamide. In some embodiments, the methods provided comprise administering to a patient in need thereof about 250 mg BID Compound 1 , about 375 mg/m ² rituximab, and about 70 mg/m ² bendamustine.

In some embodiments, the methods provided comprise administering about 250 mg of BID Compound 1 and about 500 mg/m ² of rituximab to a patient in need thereof. In some of these embodiments, the methods provided additionally comprise administering about 25 mg/m ^{2 of} fludarabine and about 250 mg/m ^{2 of} cyclophosphamide. In some embodiments, the methods provided comprise administering to a patient in need thereof about 250 mg BID Compound 1 , about 500 mg/m ² rituximab, and about 70 mg/m ² bendamustine.

In some embodiments, the methods provided comprise administering about 375 mg of BID Compound 1 and about 375 mg/m ² of rituximab to a patient in need thereof. In some of these embodiments, the methods provided additionally comprise administering about 25 mg/m ^{2 of} fludarabine and about 250 mg/m ^{2 of} cyclophosphamide. In some embodiments, the methods provided comprise administering to a patient in need thereof about 375 mg BID Compound 1 , about 375 mg/m ² rituximab, and about 70 mg/m ² bendamustine.

In some embodiments, the methods provided comprise administering to a patient in need thereof about 375 mg BID Compound 1 and about 500 mg/m ² rituximab. In some of these embodiments, the methods provided additionally comprise administering about 25 mg/m ^{2 of} fludarabine and about 250 mg/m ^{2 of} cyclophosphamide. In some embodiments, the methods provided comprise administering to a patient in need thereof about 375 mg BID Compound 1 , about 500 mg/m ² rituximab, and about 70 mg/m ² bendamustine.

In some embodiments, the methods provided comprise administering about 500 mg of BID Compound 1 and about 375 mg/m ² of rituximab to a patient in need thereof. In some of these embodiments, the methods provided additionally comprise administering about 25 mg/m ^{2 of} fludarabine and about 250 mg/m ^{2 of} cyclophosphamide. In some embodiments, the methods provided comprise administering to a patient in need thereof about 500 mg BID Compound 1 , about 375 mg/m ² rituximab, and about 70 mg/m ² bendamustine.

In some embodiments, the methods provided comprise administering about 500 mg of BID Compound 1 and about 500 mg/m ² of rituximab to a patient in need thereof. In some of these embodiments, the methods provided additionally comprise administering about 25 mg/m ^{2 of} fludarabine and about 250 mg/m ^{2 of} cyclophosphamide. In some embodiments, the methods provided comprise administering to a patient in need thereof about 500 mg BID Compound 1 , about 500 mg/m ² rituximab, and about 70 mg/m ² bendamustine.

In some embodiments, rituximab is administered once in a 28 day period. In some embodiments, rituximab is administered on day 1 or day 2 of the first cycle. In some embodiments, rituximab is administered on the first day of the 28 day cycle. In some embodiments, rituximab is administered on the first day of the second cycle. In some embodiments, rituximab is administered on the first day of the third cycle. In some embodiments, rituximab is administered on the first day of cycle 4. In some embodiments, rituximab is administered on the first day of cycle 5. In some embodiments, Rituximab was administered on the first day of cycle 6. In some embodiments, rituximab is on day 1 or 2 of the first cycle, day 1 of cycle 2, day 1 of cycle 3, day 1 of cycle 4, day 1 of cycle 5 And every day of the first day of the sixth cycle is committed.

In some embodiments, 375 mg/m ² rituximab is administered on day 1 or day ² of the first cycle, and 500 mg/m ² rituximab is administered on day 1 of the second cycle. In some embodiments, 375 mg/m ² of rituximab is administered on the first or second day of the first cycle, and 500 mg/m is administered on the first day of the second cycle and the first day of the third cycle. ² rituximab. In some embodiments, 375 mg/m ² of rituximab is administered on the first or second day of the first cycle, and the first day of the second cycle, the first day of the third cycle, and the first of the fourth cycle Each day, 500 mg/m ² of rituximab was administered. In some embodiments, 375 mg/m ² of rituximab is administered on the first or second day of the first cycle, and on the first day of the second cycle, the first day of the third cycle, and the first of the fourth cycle On the first day of the fifth and fifth cycles, 500 mg/m ² of rituximab was administered. In some embodiments, 375 mg/m ² of rituximab is administered on the first or second day of the first cycle, and on the first day of the second cycle, the first day of the third cycle, and the first of the fourth cycle On the first day of the fifth cycle, the first day of the fifth cycle, and the first day of the sixth cycle, 500 mg/m ² of rituximab was administered.

In some embodiments, 25 mg/m ^{2 of} fludarabine is administered on days 1-3 of the first, second, third, fourth, fifth, and/or sixth cycles. In some embodiments, 250 mg/m ^{2 of} cyclophosphamide is administered on days 1-3 of the first cycle, the second cycle, the third cycle, the fourth cycle, the fifth cycle, and/or the sixth cycle. In some embodiments, 70 mg/m ^{2 of} bendamrol is administered on the first, second, third, fourth, fifth, and/or first and second days of the sixth cycle. Ting.

In some embodiments, for 28 consecutive days ( "28-day cycle") administered in combination with compounds of Time 1 and the rituximab. In some embodiments, a combination of Compound 1 and rituximab is administered for two, three, four, five or six 28 day cycles. In some embodiments, a combination of Compound 1 and rituximab is administered for one, two, three, four, five, or six 28-day cycles, and Compound 1, one, two, and three are re-administered. , four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen or fourteen 28-day cycles. In some embodiments, a combination of Compound 1 and rituximab is administered to a patient for one, two, three, four, five, or six 28-day cycles, and Compound 1 is administered for a lifetime of the patient. In some embodiments, a combination of Compound 1 and rituximab is administered to a patient for one, two, three, four, five, or six 28-day cycles, and Compound 1 is additionally administered to the patient. Or one or more 28-day cycles of either rituximab. In some embodiments, a combination of Compound 1 and rituximab is administered to a patient for a lifetime.

In some embodiments, one, two, three, four, five, or six 28-day cycles of each of Compound 1 , rituximab, fludarabine, and cyclophosphamide are administered. And re-administer compound 1, one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen or fourteen A 28-day cycle.

In some embodiments, one, two, three, four, five, or six 28-day cycles of each of Compound 1 , rituximab, and bendamustine are administered and re-injected One, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen or fourteen 28-day cycles with compound 1 . .

In some embodiments, two adjacent 28 day periods may be separated by a rest period. Such rest periods may be one, two, three, four, five, six, seven or seven days, during which no compound 1 or rituximab is administered to the patient. Or both. In a preferred embodiment, two adjacent 28 day periods are continuous.

In some embodiments, the method comprises providing to a patient in need of the compound 1 in combination with rituximab of administration, wherein the patient has failed at least one prior therapy. In some embodiments, the methods provided comprise administering to a patient in need thereof, each of Compound 1 , rituximab, fludarabine, and cyclophosphamide, wherein the patient has failed at least one previous therapy. In some embodiments, a method is provided comprising administering to a patient in need thereof, each of Compound 1 , Rituximab, and bendamustine, wherein the patient has failed at least one prior therapy.

In some embodiments, the invention provides a system for treating, stabilizing or ameliorating the severity of one or more diseases or conditions associated with BTK, the system comprising Compound 1 and rituximab. 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 rituximab.

In some embodiments, the kit comprises twenty eight (28) daily doses of Compound 1 and one 10 mg/mL vial of rituximab. In some embodiments, the kit comprises twenty eight (28) daily doses of Compound 1 and one 100 mg/10 mL vial of rituximab. In some embodiments, the kit comprises twenty eight (28) daily doses of Compound 1 and one 500 mg/50 mL vial of rituximab.

In some embodiments, the kit comprises fifty-six (56) times a 375 mg dose of Compound 1 and a 10 mg/mL vial of rituximab. In some embodiments, the kit comprises fifty-six (56) times a 375 mg dose of Compound 1 and a 100 mg/10 mL vial of rituximab. In some embodiments, the kit comprises fifty-six (56) times a 375 mg dose of Compound 1 and a 500 mg/50 mL vial of rituximab.

In some embodiments, the kit comprises two 375 mg doses of Compound 1 and one 10 mg/mL vial of rituximab. In some embodiments, the kit comprises two 375 mg doses of Compound 1 and one 100 mg/10 mL vial of rituximab. In some embodiments, the kit comprises two 375 mg doses of Compound 1 and one 500 mg/50 mL vial of rituximab. In some embodiments, the kit comprises two 500 mg doses of Compound 1 and a single 10 mg/mL dose of rituximab. In some embodiments, the kit comprises two 500 mg doses of Compound 1 and one 100 mg/10 mL vial of rituximab. In some embodiments, the kit comprises a dose of 500mg twice of Compound 1 and a 500mg / 50mL 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 generally promote drug release and absorption 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 sliding 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 rituximab to a patient in need thereof . The besylate of the compound 1 is N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilinyl)pyrimidin-4-ylamino)phenyl)acrylamide 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 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 The patient is administered each of Compound 1 , rituximab, fludarabine, and cyclophosphamide, 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 , rituximab and bendamustine, 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 patients administered the compound comprising the besylate salt of a pharmaceutical composition of a combination of rituximab, 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 of a compound of benzenesulfonate (besylate i.e. compound 1), each of rituximab, fludarabine and cyclophosphamide in the pharmaceutical composition wherein the compound The amount of the besylate salt of 1 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 . 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 of a compound of benzenesulfonate (besylate i.e. compound 1), each of rituximab and bendamustine Ting in the pharmaceutical composition of 1 wherein the compound benzenesulfonamide The amount of the acid 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 I 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% (8.30mg) intragranular; 0.5% (2.08mg) extragranular.

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 2 below, in the form of an infusion of 1 mg/mL to 4 mg/mL.

Research design

Individuals with relapsed or refractory CLL or SLL who had failed at least one previous treatment regimen in a "3+3" dose escalation and expansion study were selected to determine the non-tolerant dose of Compound 1 in combination with rituximab ( 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 oxime was administered according to the group listed in Table 2. Aminobenzenesulfonate and rituximab:

Within each group, the individual used N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamine during the first two 28-day treatment cycles Phenyl) phenyl decyl benzene sulfonate PO (oral) BID (daily) treatment and assessment of safety, tolerability and DLT, as well as 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.

Rituximab is administered as a single intravenous (IV) infusion. The initial infusion in the first cycle was administered at 375 mg/m ² ; the subsequent infusion in the second cycle to the sixth cycle was administered at 500 mg/m ² . Rituximab was administered on the second day of the first cycle and on the first day of each subsequent cycle. After the infusion of the sixth cycle, rituximab will be discontinued. If appropriate, the individual can continue to use N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl)acrylamidobenzene Sulfate treatment. The first infusion of rituximab was 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 in increments of 100 mg/h over a 30 minute interval to a maximum of 400 mg/h.

The patient registration dose should be based on the group that was opened at the time of registration. 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. If <1 of the 3 registered DLT evaluable individuals suffered DLT during the first 2 cycles, the dose should be considered lower than NTD. When at least two (2) of the six (6) DLT evaluable individuals in the group are subjected 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 has suffered DLT during the first two 28 day periods.

During the dose escalation period, the judgment of the next higher dose group should be based on the safety and review of the DLT evaluable patient. The OBE dose is defined as follows: ● Six (6) individuals Two (2) lymph node size reduction 50%; and/or ● no additional exposure increases with dose; and/or ● increased lymphocytosis in four (4) of six (6) individuals during the first three 28-day cycles 25% were not assessed as progressive disease.

Results. "Complete response" (CR) according to IWCLL guidelines, 2008 (no LN>1.5cm, 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. 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.

Three individuals were registered in Group 1 and N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl)propene oxime Combination treatment with amphetamine and rituximab. Of the three individuals enrolled in Group 1 on October 16, 2013, one individual is currently in the ninth 28-day cycle and two individuals are currently in the eighth 28-day cycle. Since none of these individuals suffered DLT during the first 28-day period, three individuals were registered in group 2. Two of the three individuals in Group 2 ended the first two 28-day cycles without DLT. The third individual has suffered a grade 3 fatigue and weak DLT graded by the NCI-CTCAE guidelines. The other three individuals were enrolled in Group 2 for further assessment of DLT and safety of this dose.

Figure 1 summarizes patient registration and response assessment. All three patients in Group 1 have been evaluated as having a partial response to the study treatment. In all three cases, the investigator's PR assessment by the third cycle was confirmed by a CT scan showing a reduction in lymph node size of more than 50% compared to the pre-treatment baseline values. The first individual registered is currently in the 9th cycle of treatment and continues to maintain partial response. The other two patients in Group 1 had continued treatment into Cycle 8 and also maintained partial response to treatment. Of the six individuals treated in Group 2, two individuals had achieved partial response before the start of Cycle 3, according to the investigator's test assessment. According to the investigator's assessment, an individual has shown partial response by the fifth cycle. The assessment of the response of the other 3 individuals enrolled in Group 2 is pending.

All dose escalation groups are registered and each dose escalation group ends at the second treatment cycle After that, the number and type of DLTs and adverse events (AEs) that occurred during the first two cycles will be assessed. 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.

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.

During the expansion of the group, rituximab was administered according to the time course stated in the dose escalation group. 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 . Rituximab was administered on the second day of the first cycle and on the first day of each subsequent cycle. After the infusion of the sixth cycle, rituximab was discontinued. Each body will continue to use N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl)acrylamidobenzenesulfonic acid Salt treatment until disease progression, unacceptable toxicity, or treatment discontinuation for any other reason.

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 two 28-day cycles (approximately 56 days) and considered to be treatment-related. Hematology DLT includes grade 4 anemia (reduced heme) or thrombocytopenia according to NCI-CTCAE (version 4.03) or IWCLL criteria, either causing a drop in blood threshold; grade 4 neutropenia over 5 days, Even with granulocyte community stimulating factor (G-CSF) support; and grade 3 or higher febrile neutropenia. Lymphocytosis can be observed as a result of disease progression, but has also been described as a phenomenon of redistribution (lymphocyte migration and transport) in individuals receiving another BTK inhibitor, even as a lymph node disease that responds to treatment. Therefore, lymphocytosis will not be included in the DLT. Malignant lymphocytosis is a prescribed therapeutic effect of treatment and is not considered DLT.

Non-hematologic DLT includes non-hematologic AEs of grade 4 or higher with any duration; elevated grade 3 bilirubin, whether symptomatic or asymptomatic; and less than 24 hours of nausea after medical therapy, Any grade 3 non-hematologic toxicity other than vomiting and diarrhea; tumor lysis syndrome that has not progressed to grade 4 and is medically eliminated within 7 days is not considered DLT; and transient, asymptomatic and rapidly reversible (within 7 days) Back to baseline or Level 3) Level 3 non-hematology laboratory abnormalities are not considered DLT.

Individuals without disease progression at the end of the first two 28-day treatment cycles and without DLT are eligible to continue receiving N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidine Combination of -4-ylamino)phenyl)propenylamine besylate with rituximab for a further 28-day period until (i) the patient suffers from unacceptable toxicity and (ii) the progression of the underlying malignancy (iii) the patient withdraws consent, or (iv) the treating physician investigator otherwise determines that the patient should not continue treatment. If the treatment investigator determines that the individual suffering from the DLT has a clinical benefit from the study treatment, the individual may continue to conduct the study treatment. Rituximab was administered only for the first 6 cycles; however, it continued to benefit from N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidine-4- Patients with amino)phenyl)acrylamidobenzenesulfonate can continue to be treated.

One of the individuals in Group 1 suffered from herpetic esophagitis in the second cycle and suffered two SAEs in the fifth cycle; atrial fibrillation and pneumonia episodes, and hypotension, mental state changes, and pneumonia episodes, two reports Both are considered to be unrelated to the study drug. All three of these SAE reports required brief hospitalization and drug interruption, but after re-attacking with the drug, the individual subsequently achieved further improvement in disease status. In group 2, one individual suffered a scrotal abscess during the second cycle, which was considered unrelated to the study drug. Another group of 2 individuals suffered from grade 3 fatigue during the second cycle, which was declared DLT; however, the AE was not reported as severe. This patient takes a reduced dose of N- (3-(5-fluoro-2-(4-(2-methoxyethoxy)anilino)pyrimidin-4-ylamino)phenyl)propenyl benzene sulfonate The acid salt continues to be studied and treated. Another individual in Group 2 suffered from visual impairment during the second cycle, which was present at baseline. Two individuals (one in each group) reported grade 3 neutropenia.

Example 2

Screening 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 mode with 3 fold serial dilutions starting at 1 μM, 50 μM or 100 μM. Positive control. 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 3 illustrates the average percent inhibition of Compound 2 against various kinases:

Example 3

This study were to include N - (3- (5- fluoro-2- (4- (2-methoxyethoxy) phenylamino) pyrimidin-4-ylamino) phenyl) acrylamide benzenesulfonic The capsule of the salt (Compound 1 benzenesulfonate) corresponds to Table 1 in 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 4 below, at a dose of 1 mg/mL to 4 mg/mL.

Fludarabine is approved as Fludara® and is commercially available in vials containing sterile freeze-dried solid cakes containing 50 mg of fludarabine phosphate, 50 mg of mannitol and sodium hydroxide. The solid cake was reconstituted with 2 mL of sterile water for injection USP to yield a 25 mg/mL solution.

Cyclophosphamide is approved as Cytoxan® and is commercially available as a sterile powder, which can be reconstituted according to the instructions for the drug.

Bendamustine is approved as Treanda® and is commercially available as a single-use vial containing 100 mg of bendamustine hydrochloride as a lyophilized powder. The powder was reconstituted with 20 mL of sterile water for injection USP to yield a 5 mg/mL solution which was further diluted with 0.9% sodium chloride injection USP or 2.5% dextrose/0.45% sodium chloride injection USP immediately prior to injection (final concentration) It is 0.2-0.6 mg/mL).

Research design

Individuals with relapsed or refractory CLL or SLL who had failed at least one previous treatment regimen in the "3+3" dose escalation and expansion study to determine Compound 1 , Rituximab, Fludarabine, and Cyclophosphamide Non-tolerant dose (NTD), optimal biological effect dose (OBE) and maximum tolerated dose (MTD) of indoleamine (group A) and compound 1 , rituximab and bendamustine (group B) . Approximately 30-42 patients are expected to be enrolled in the study.

The study treatment will be 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 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.

Compounds 1 besylate, rituximab, fludarabine, and cyclophosphamide were administered according to the groups in Group A studies listed in Table 4:

Compounds 1 besylate, rituximab, and bendamustine were administered according to the groups in Group B studies listed in Table 5:

Within each group, the individual will treat BID (daily) with Compound 1 besylate PO (oral) according to Table 4 or 5 during two initial 28-day treatment cycles and will assess safety, tolerability and DLT, And pharmacokinetics ("PK"), pharmacodynamics ("PD") and disease response.

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 . Rituximab will be administered on the first day of the first cycle and on the first day of each subsequent 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. 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.

On days 1-3 of the 1-6th cycle, 25 mg/m ² fludarabine was administered as an intravenous infusion of 25 mg/mL (sterile water USP) over 20-30 minutes. On days 1-3 of the first to sixth cycles, 250 mg/m ^{2 of} cyclophosphamide was administered as an intravenous infusion of 100 mg/5 mL over 10-30 minutes.

Bendamustine was diluted with sterile water for injection to a concentration of 5 mg/mL. Prior to use, the bendamustine solution should be immediately transferred to a 500 mL infusion bag with 0.9% sodium chloride injection USP. Next, on day 1 and day 2 of the 1-6th cycle, 70 mg/m ² of the bendamustine infusion solution was administered as an intravenous infusion over 30-60 minutes.

Example 4

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 the salt (Compound 1 benzenesulfonate) corresponds to Table 1 in Example 1.

Various doses of orfarizumab were prepared in 1000 mL of 0.9% sodium chloride injection USP solution prior to administration. Olfazumab is diluted as follows:

● 300mg dose: 5mL from each of the 3 single use 100mg vials To 985mL 0.9% sodium chloride injection USP.

• 2000 mg dose: 50 mL to 900 mL of 0.9% sodium chloride injection USP was added from each of 2 single use 1000 mg vials.

Research design

Individuals with relapsed or refractory CLL or SLL who have failed at least one previous treatment regimen in the "3+3" dose escalation and expansion study will be tested to determine the non-tolerant dose of Compound 1 and orfarizumab (NTD) ), optimal biological effect dose (OBE) and maximum tolerated dose (MTD). Approximately 30-42 patients are expected to be enrolled in the study.

The study treatment will be 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 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.

Compound 1 besylate and orfarizumab will be administered according to the group for study listed in Table 6:

Within each group, the individual will treat BID (daily) with Compound 1 besylate PO (oral) according to Table 6 during two initial 28-day treatment cycles and will assess safety, tolerability and DLT, as well as drugs Kinetics ("PK"), pharmacodynamics ("PD") and disease response.

Orfarizumab will be administered as a single intravenous (IV) infusion. The initial infusion during the first dose should be at a rate of 3.6 mg/hr (12 ml/hr). The infusion rate of dose 2 should be at a rate of 24 mg/hr (12 ml/hr). The subsequent infusion rate should be 50 mg/hr (25 ml/hr). In the absence of infusion toxicity, the infusion rate can be increased every 30 minutes as described in Table 7: Table 7. Infusion rate of olfazumab

^a dose 1 = 300mg (0.3mg / mL)

^b dose 2 and 3-12 = 2000mg (2mg/mL)

Claims (38)

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 an anti-CD20 antibody. 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 the patient has failed at least one prior therapy. The method of requesting the item 2 or 3, wherein the compound 1 administered twice one day. The method of claim 5, wherein the compound 1 is in the form of a besylate salt. The method of claim 6, 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 7, 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 6, wherein the compound 1 is administered in an oral dosage form. The method of claim 2 or 3, wherein the anti-CD20 antibody is rituximab. The method of claim 2 or 3, wherein the anti-CD20 antibody is ofofumumab. The method of claim 10, wherein the rituximab is administered once in a 28 day period. The method of claim 12, wherein the rituximab is administered as an intravenous infusion. The method of claim 10, wherein Compound 1 and rituximab are each administered for at least one 28 day period. The method of claim 2 or 3, wherein the method additionally comprises administering fludarabine and cyclophosphamide. The method of claim 2 or 3, wherein the method additionally comprises administering bendamustine. 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 and a therapeutically effective amount of an anti-CD20 antibody, wherein the therapeutically effective amount of Compound 1 is from about 750 mg to about 1000 mg per day. The method of claim 17, wherein the therapeutically effective amount of Compound 1 is about 375 mg BID. The method of claim 17, wherein the therapeutically effective amount of Compound 1 is about 500 mg BID. The method of claim 17, wherein the anti-CD20 antibody is rituximab. The method of claim 17, wherein the anti-CD20 antibody is orfarizumab. The method of claim 20, wherein the therapeutically effective amount of rituximab is about 375 mg/m ² . The method of claim 20, wherein the therapeutically effective amount of rituximab is about 500 mg/m ² . The method of claim 17, wherein the method additionally comprises administering fludarabine and cyclophosphine Guanamine. The method of claim 17, wherein the method additionally comprises administering bendamustine. A system for treating, stabilizing or attenuating the severity of one or more BTK-associated diseases or conditions comprising Compound 1 and an anti-CD20 antibody. The system of claim 26, wherein the anti-CD20 antibody is rituximab. The system of claim 26, wherein the anti-CD20 antibody is orfarizumab. The system of claim 26 additionally comprises fludarabine and cyclophosphamide. The system of claim 26, additionally comprising bendamustine. 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 and an anti-CD20 antibody to a patient in need thereof, wherein the irreversible BTK inhibitor has an option No more than about 50% inhibition of kinases from: 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 31, 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 31, 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 31, wherein the irreversible BTK inhibitor has a percent inhibition of LYN of no more than about 20-30%. The method of claim 31, wherein the anti-CD20 antibody is rituximab. The method of claim 31, wherein the anti-CD20 antibody is orfarizumab. The method of claim 31, wherein the method additionally comprises administering fludarabine and cyclophosphine Guanamine. The method of claim 31, wherein the method additionally comprises administering bendamustine.