HCK as a therapeutic target for MYD88 mutation diseases

Abstract

Provided herein are methods for treating a disease (e.g., a proliferative disease (e.g., a cancer (e.g., breast cancer, colon cancer, testicular cancer, CNS cancer, gastric cancer, lymphoma (e.g., B cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgM-secreting lymphoplasmacytic lymphoma (i.e., Waldenstrom's macroglobulinemia), non-IgM-secreting lymphoplasmacytic lymphoma)), diffuse large B cell lymphoma (e.g., activated B cell-like (ABC)-DLBCL, germinal center B cell-like (GBC)-DLBCL), follicular lymphoma, marginal zone B cell lymphoma, small lymphocytic lymphoma, mantle cell lymphoma) and leukemia (e.g., chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia, myeloid leukemia (e.g., chronic myeloid leukemia, acute myeloid leukemia))))) comprising administering to a subject in need thereof a therapeutically effective amount of a compound (I). Further provided are methods for treating a disease that is resistant to treatment with a BTK inhibitor (eg, ibrutinib).

Description

[0001] Related Applications

[0002] This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Application USSN 62 / 912,474, filed on October 8, 2019, which is incorporated herein by reference. Background Art

[0003] It has been found that hematopoietic cell kinase (HCK) transcription and activation is triggered by mutant myeloid differentiation primary response 88 (MYD88) and is an important determinant of pro-survival signaling. It has also been found that inhibition of HCK kinase activity triggers apoptosis of mutant MYD88 cells. For example, MYD88 mutations are found in Waldenstrom's macroglobulinemia ( Macroglobulinemia (WM), in which 95 to 97% of patients express MYD88 L265P , while MYD88 mutations other than L265P are rare. WM is considered to correspond to lymphoplasmacytic lymphoma (LPL) as defined by the World Health Organization classification system. Up to 30% of patients with the activated B-cell (ABC) subtype of diffuse large B-cell lymphoma (ABC DLBCL) also express activating MYD88 mutations, including MYD88 L265P Mutations in MYD88 promote Myddosome self-assembly and can trigger NF-kB signaling via IL1 receptor-associated kinase (IRAK4 / IRAK1) or Bruton's tyrosine kinase (BTK) in the absence of Toll (TLR) or IL1 (IL1R) receptor signaling.

[0004] Next generation sequencing has revealed activated myeloid differentiation primary response 88 (MYD88) mutations in several B-cell malignancies, including Waldenstrom's macroglobulinemia (lymphoplasmacytic lymphoma that secretes immunoglobulin M (IgM)), non-IgM-secreting lymphoplasmacytic lymphoma, the ABC subtype of diffuse large B-cell lymphoma, primary central nervous system (CNS) lymphoma, immune-privileged lymphomas including testicular lymphoma, marginal zone lymphoma, and chronic lymphocytic leukemia. Particularly striking is the expression of MYD88 mutations in Waldenstrom's macroglobulinemia (WM), in which 95 to 97% of patients express MYD88. L265P , while non-L265P MYD88 mutations are rare. Waldenstrom's macroglobulinemia is considered to correspond to lymphoplasmacytic lymphoma (LPL) as defined by the World Health Organization classification system. Up to 30% of patients with the activated B-cell (ABC) subtype of diffuse large B-cell lymphoma (ABC-DLBCL) also express activating MYD88 mutations, including MYD88 L265PMutations in MYD88 promote Myddosome self-assembly and can trigger NF-kB signaling via IL1 receptor-associated kinase (IRAK4 / IRAK1) or Bruton's tyrosine kinase (BTK) in the absence of Toll (TLR) or IL1 (IL1R) receptor signaling.

[0005] Ibrutinib (IB) is a BTK inhibitor with high activity in WM, producing responses in 91% of previously treated patients. In WM patients, patients with MYD88 mutations had higher primary and overall responses to ibrutinib. Ibrutinib also showed activity in previously treated ABC DLBCL patients, particularly in patients with MYD88 mutations. Ibrutinib is also active in other B-cell malignancies including chronic lymphocytic leukemia (CLL) and mantle cell lymphoma (MCL). Inhibition of tonic B cell receptor (BCR) activity mediated by BTK has been suggested as a potential mechanism of ibrutinib activity in non-WM B-cell diseases. Summary of the invention

[0006] Provided herein are methods for treating a disease (e.g., a proliferative disease (e.g., an IgM gammopathy (e.g., IgM monoclonal gammopathy of undetermined significance (MGUS), amyloid light chain (AL) amyloidosis), a mastocytosis (e.g., systemic mastocytosis), a cancer (e.g., breast cancer, colon cancer, testicular cancer, CNS cancer, gastric cancer, a lymphoma (e.g., a B-cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgM-secreting lymphoplasmacytic lymphoma (i.e., Waldenstrom's macroglobulinemia), a non-IgM-secreting lymphoplasmacytic lymphoma)), a diffuse large B-cell lymphoma (e.g., activated B A method for treating a subject with a compound of formula (I) comprising administering to the subject a therapeutically effective amount of:

[0007]

[0008] or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, isotope-labeled derivative, stereoisomer or prodrug thereof.

[0009] Compound (I) can be a kinase inhibitor (e.g., SRC family kinase (i.e., SFK) (e.g., HCK, LYN, BLK, FRK), Tec family kinase (e.g., BTK)), and in certain aspects, the compound can be specific or selective for SFK (e.g., HCK, LYN, BLK, FRK) or Tec family kinase (e.g., BTK) relative to one or more other kinases. Pharmaceutical compositions and kits comprising compound (I) are also provided. The present disclosure also provides methods of using the disclosed compounds, pharmaceutical compositions and kits (e.g., for treating a disease in a subject in need thereof (e.g., a proliferative disease (e.g., IgM gammopathy (e.g., IgM monoclonal gammopathy of undetermined significance (MGUS), amyloid light chain (AL) amyloidosis), mastocytosis (e.g., systemic mastocytosis), cancer (e.g., breast cancer, colon cancer, testicular cancer, CNS cancer, gastric cancer, lymphoma (e.g., B cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgM secreting lymphoplasmacytic lymphoma (i.e., Waldenstrom's macroglobulinemia), non-IgM secreting lymphoplasmacytic lymphoma)), diffuse Large B cell lymphoma (e.g., activated B cell-like (ABC)-DLBCL, germinal center B cell-like (GBC)-DLBCL), follicular lymphoma, marginal zone B cell lymphoma, small lymphocytic lymphoma, mantle cell lymphoma), myeloma (e.g., IgM myeloma (e.g., IgM multiple myeloma)) and leukemia (e.g., chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia, myeloid leukemia (e.g., chronic myeloid leukemia, acute myeloid leukemia (e.g., mast cell leukemia), myeloproliferative disease (e.g., myelodysplastic syndrome))))), or inhibiting kinase activity in a subject, biological sample or cell in need thereof).

[0010] In yet another aspect, the present disclosure provides compound (I), and pharmaceutically acceptable salts, solvates, hydrates, polymorphs, cocrystals, tautomers, stereoisomers, isotope-labeled derivatives and prodrugs thereof, and pharmaceutical compositions thereof, for use in treating and / or preventing a disease (e.g., a proliferative disease such as IgM gammopathy, mastocytosis, cancer) in a subject in need thereof.

[0011] In another aspect, the present disclosure provides compound (I), and pharmaceutically acceptable salts, solvates, hydrates, polymorphs, cocrystals, tautomers, stereoisomers, isotope-labeled derivatives and prodrugs thereof, and pharmaceutical compositions thereof for use in the preparation of a medicament for treating and / or preventing a disease in a subject in need thereof.

[0012] In another aspect, the present disclosure provides a method for preparing Compound (I) or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, tautomer, stereoisomer, isotopically labeled derivative or prodrug thereof.

[0013] In one aspect, the present disclosure provides a pharmaceutical composition comprising compound (I) and an optional pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition further comprises another medicament. In certain embodiments, another medicament is selected from the group consisting of chemotherapeutics, epigenetic modifiers, glucocorticoids, biologics, and immunotherapeutics. On the other hand, another medicament is a BCL-2 inhibitor (e.g., venetoclax, navitoclax, obatoclax).

[0014] The pharmaceutical composition can be used to treat a disease in a subject in need, inhibiting kinase activity in a subject in need, a biological sample or a cell. In certain aspects, the disease is a proliferative disease (e.g., IgM gammopathy (e.g., IgM monoclonal gammopathy of undetermined significance (MGUS), amyloid light chain (AL) amyloidosis), mastocytosis (e.g., systemic mastocytosis), cancer (e.g., breast cancer, colon cancer, testicular cancer, CNS cancer, gastric cancer, lymphoma (e.g., B cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgM secreting lymphoplasmacytic lymphoma (i.e., Waldenstrom's macroglobulinemia), non-IgM secreting lymphoplasmacytic lymphoma)), diffuse large B cell lymphoma (e.g., activated B-cell-like (ABC)-DLBCL, germinal center B-cell-like (GBC)-DLBCL), follicular lymphoma, marginal zone B-cell lymphoma, small lymphocytic lymphoma, mantle cell lymphoma), myeloma (e.g., IgM myeloma (e.g., IgM multiple myeloma)), and leukemia (e.g., chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia, myeloid leukemia (e.g., chronic myeloid leukemia, acute myeloid leukemia (e.g., mast cell leukemia), myeloproliferative disorders (e.g., myelodysplastic syndrome))))).

[0015] The present disclosure provides a method for treating a disease in a subject by administering an effective amount of a compound (I) as described herein or a pharmaceutical composition thereof to a subject in need thereof. In certain aspects, the disease is a proliferative disease (e.g., IgM gammopathy (e.g., IgM monoclonal gammopathy of undetermined significance (MGUS), amyloid light chain (AL) amyloidosis), mastocytosis (e.g., systemic mastocytosis), cancer (e.g., breast cancer, colon cancer, testicular cancer, CNS cancer, gastric cancer, lymphoma (e.g., B cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgM secreting lymphoplasmacytic lymphoma (i.e., Waldenstrom's macroglobulinemia), non-IgM secreting lymphoplasmacytic lymphoma)), diffuse large B cell lymphoma (e.g., activated B cell-like (ABC)-DLBCL, germinal center B cell-like (GBC)-DLBCL), follicular lymphoma, marginal zone B cell lymphoma, small lymphocytic lymphoma, mantle cell lymphoma), myeloma (e.g., IgM myeloma (e.g., IgM multiple myeloma)) and leukemia (e.g., chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia, myeloid leukemia (e.g., chronic myeloid leukemia, acute myeloid leukemia (e.g., mast cell leukemia), myeloproliferative disease (e.g., myelodysplastic syndrome))))). Methods for contacting a biological sample or cell with an effective amount of a compound as described herein or a pharmaceutical composition thereof are also described. In certain embodiments, the methods described herein further comprise administering an additional agent to a subject in need thereof. In certain embodiments, the methods described herein further comprise contacting a biological sample or cell with an additional agent.

[0016] In one aspect, a method for treating a disease in a subject resistant to treatment with a BTK inhibitor (e.g., ibrutinib, CC-292, ONO-4059, evobrutinib, spebrutinib, BGB-3111, HM71224, or ACP-196 (i.e., acalabrutinib)) is provided. In certain aspects, a subject resistant to treatment with a BTK inhibitor has a mutated BTK (e.g., a C481S mutated BTK). In certain aspects, a subject has a C481S mutated BTK. On the other hand, a subject resistant to treatment with a BTK inhibitor has a C481S mutated BTK. On the other hand, a subject resistant to treatment with a BTK inhibitor is diagnosed as having a MYD88 mutation disease (e.g., a proliferative disease (e.g., IgM gammopathies, mastocytosis, or cancer)).

[0017] On the other hand, provided herein is a method of inhibiting a kinase (e.g., SFK (e.g., HCK, LYN, BLK, FRK), Tec family kinase (e.g., BTK)) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of Compound (I) or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, isotopically labeled derivative, stereoisomer or prodrug thereof. In certain aspects, provided herein is a method of inhibiting a mutant BTK (e.g., C481S mutant) in a subject in need thereof.

[0018] In certain aspects, the method further comprises administering an anticancer agent to the subject. In some embodiments, the anticancer agent is a chemotherapeutic agent. In another aspect, the method further comprises administering to the subject a proteasome inhibitor (e.g., bortezomib, carfilzomib, ixazomib, or oprozomib), a monoclonal antibody (e.g., rituximab, daratumumab, ofatumumab, or obinituzumab), an alkylating agent (e.g., bendamustine), or a combination thereof. ), cyclophosphamide), nucleoside analogs (e.g., fludarabine or cladribine), mTOR inhibitors (e.g., everolimus), BTK inhibitors (e.g., ibrutinib, acalabrutinib or BGB-3111), BCR inhibitors (e.g., SYK inhibitors) and / or immunomodulators (e.g., thalidomide or lenalidomide).

[0019] In another aspect, the present disclosure provides a kit comprising: Compound (I), or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, tautomer, stereoisomer, isotope-labeled derivative or prodrug thereof, or a pharmaceutical composition thereof; and

[0020] Instructions for use of the compound or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, tautomer, stereoisomer, isotopically labeled derivative or prodrug or pharmaceutical composition thereof.

[0021] The details of one or more embodiments of the present disclosure are set forth herein. Other features, objects, and advantages of the present disclosure will be apparent from the detailed description, examples, drawings, and claims.

[0022] definition

[0023] Definitions of specific functional groups and chemical terms are described in more detail below.Chemical elements are identified according to the Periodic Table of the Elements (CAS version, Handbook of Chemistry and Physics, 75th ed., inside cover), and specific functional groups are generally defined as described. In addition, general principles of organic chemistry as well as specific functional moieties and reactivities are described in Thomas Sorrell, Organic Chemistry, University Science Books, Sausalito, 1999; Michael B. Smith, March's Advanced Organic Chemistry, 7th ed., John Wiley & Sons, Inc., New York, 2013; Richard C. Larock, Comprehensive Organic Transformations, John Wiley & Sons, Inc., New York, 2018; and Carruthers, Some Modern Methods of Organic Synthesis, 3rd ed., Cambridge University Press, Cambridge, 1987.

[0024] The compounds described herein may include one or more asymmetric centers and may therefore exist in various stereoisomeric forms, such as enantiomers and / or diastereomers. For example, the compounds described herein may be in the form of individual enantiomers, diastereomers, or geometric isomers, or may be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers may be separated from the mixture by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers may be prepared by asymmetric synthesis. See, e.g., Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, EL, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, SH, Tables of Resolving Agents and Optical Resolutions, p. 268 (Eliel, ed., Univ. of Notre Dame Press, Notre Dame, IN, 1972). The present disclosure additionally encompasses the compounds described herein as individual isomers substantially free of other isomers, and alternatively as mixtures of various isomers.

[0025] When a numerical range ("range") is listed, it includes every value and subrange within the range. Unless otherwise specified, a range includes values ​​at both ends of the range. For example, "C 1–6 "Alkyl" covers C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 1–6 , C 1–5 , C 1–4 , C 1–3 , C 1–2 , C 2–6 , C 2–5 , C2–4 , C 2–3 , C 3–6 , C 3–5 , C 3–4 , C 4–6 , C 4–5 and C 5–6 alkyl.

[0026] "Hydrocarbon chain" refers to a substituted or unsubstituted divalent alkyl, alkenyl, or alkynyl group. A hydrocarbon chain comprises (1) one or more carbon atom chains between two groups immediately adjacent to the hydrocarbon chain; (2) optionally, one or more hydrogen atoms on the carbon atom chain; and (3) optionally, one or more substituents on the carbon atom chain ("non-chain substituents", which are not hydrogen). The carbon atom chain consists of continuously linked carbon atoms ("chain atoms") and does not contain hydrogen atoms or heteroatoms. However, the non-chain substituents of a hydrocarbon chain may contain any atom, including hydrogen atoms, carbon atoms, and heteroatoms. For example, the hydrocarbon chain -C A H(C B H 2 C C H 3 ) – contains a chain atom C A , C A A hydrogen atom and a non-chain substituent on the B H 2 C C H 3 ). The term “C x A hydrocarbon chain", where x is a positive integer, refers to a hydrocarbon chain containing x chain atoms between the two radicals of the hydrocarbon chain. If there is more than one possible value for x, the smallest possible value of x is used to define the hydrocarbon chain. For example, –CH(C 2 H 5 ) – is C 1 hydrocarbon chain, and It is C 3 When a numerical range is used, the meaning of the range is as described herein. For example, C 3-10 A hydrocarbon chain is one in which the number of carbon atoms in the shortest chain between two groups adjacent to the hydrocarbon chain is 3, 4, 5, 6, 7, 8, 9, or 10. The hydrocarbon chain may be saturated (e.g., –C≡C– or –(CH 2 ) 4 –). The hydrocarbon chain may also be unsaturated and contain one or more C=C and / or C≡C bonds at any position in the hydrocarbon chain. For example, –CH=CH– (CH 2 ) 2 –, –CH 2 –C≡C–CH 2 – and –C≡C–CH=CH– are examples of unsubstituted and unsaturated hydrocarbon chains. In certain embodiments, the hydrocarbon chain is unsubstituted (e.g., –(CH 2) 4 In certain embodiments, the hydrocarbon chain is substituted (e.g., -CH(C 2 H 5 )– and –CF 2 –). Any two substituents on the hydrocarbon chain may be linked to form an optionally substituted carbocyclic, optionally substituted heterocyclic, optionally substituted aryl or optionally substituted heteroaryl ring. For example, are all examples of hydrocarbon chains. In contrast, in certain embodiments, Not within the scope of the hydrocarbon chain described herein. x When a hydrocarbon chain atom is replaced by a heteroatom, the resulting group is called a C x Hydrocarbon chains in which chain atoms are replaced by heteroatoms, with C x-1 The hydrocarbon chains are opposite. For example, is a C in which one of the chain atoms is replaced by an oxygen atom 3 Hydrocarbon chain.

[0027] The term "alkyl" refers to a straight-chain or branched saturated hydrocarbon group having 1 to 20 carbon atoms ("C 1–20 In some embodiments, an alkyl group has 1 to 12 carbon atoms (“C 1–12 In some embodiments, an alkyl group has 1 to 10 carbon atoms (“C 1-10 In some embodiments, an alkyl group has 1 to 9 carbon atoms (“C 1-9 In some embodiments, an alkyl group has 1 to 8 carbon atoms (“C 1-8 In some embodiments, an alkyl group has 1 to 7 carbon atoms (“C 1-7 In some embodiments, an alkyl group has 1 to 6 carbon atoms (“C 1-6 In some embodiments, an alkyl group has 1 to 5 carbon atoms (“C 1-5 In some embodiments, an alkyl group has 1 to 4 carbon atoms (“C 1-4 In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C 1-3 In some embodiments, an alkyl group has 1 to 2 carbon atoms (“C 1-2 In some embodiments, an alkyl group has 1 carbon atom (“C 1 In some embodiments, the alkyl group has 2 to 6 carbon atoms (“C 2–6 C 1–6 Examples of alkyl groups include methyl (C 1 ), ethyl (C 2 ), propyl (C 3) (e.g., n-propyl, isopropyl), butyl (C 4 ) (e.g., n-butyl, tert-butyl, sec-butyl, isobutyl), pentyl (C 5 ) (e.g., n-pentyl, 3-pentyl, pentyl, neopentyl, 3-methyl-2-butyl, tert-pentyl) and hexyl (C 6 ) (e.g., n-hexyl). Other examples of alkyl groups include n-heptyl (C 7 ), n-octyl (C 8 ), n-dodecyl (C 12 ) etc. Unless otherwise specified, each instance of an alkyl group is independently unsubstituted (an “unsubstituted alkyl group”) or substituted (a “substituted alkyl group”) with one or more substituents (e.g., halogen, such as F). In certain embodiments, an alkyl group is an unsubstituted C 1–12 Alkyl (such as unsubstituted C 1–6 Alkyl groups, such as –CH 3 (Me), unsubstituted ethyl (Et), unsubstituted propyl (Pr, for example, unsubstituted n-propyl (n-Pr), unsubstituted isopropyl (i-Pr)), unsubstituted butyl (Bu, for example, unsubstituted n-butyl (n-Bu), unsubstituted tert-butyl (tert-Bu or t-Bu), unsubstituted sec-butyl (sec-Bu or s-Bu), unsubstituted isobutyl (i-Bu)). In certain embodiments, the alkyl group is substituted C 1–12 Alkyl (such as substituted C 1–6 Alkyl groups, such as –CH 2 F, –CHF 2 , –CF 3 , –CH 2 CH 2 F, –CH 2 CHF 2 , –CH 2 CF 3 or benzyl (Bn).

[0028] The term "alkenyl" refers to a straight or branched hydrocarbon group having 2 to 20 carbon atoms and one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 double bonds). In some embodiments, the alkenyl group has 2 to 20 carbon atoms ("C 2–20 In some embodiments, an alkenyl group has 2 to 12 carbon atoms (“C 2-12 In some embodiments, an alkenyl group has 2 to 11 carbon atoms (“C 2-11 In some embodiments, an alkenyl group has 2 to 10 carbon atoms (“C 2-10 In some embodiments, an alkenyl group has 2 to 9 carbon atoms (“C 2-9In some embodiments, an alkenyl group has 2 to 8 carbon atoms (“C 2-8 In some embodiments, an alkenyl group has 2 to 7 carbon atoms (“C 2-7 In some embodiments, an alkenyl group has 2 to 6 carbon atoms (“C 2-3 In some embodiments, an alkenyl group has 2 to 5 carbon atoms (“C 2-5 In some embodiments, an alkenyl group has 2 to 4 carbon atoms (“C 2-4 In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C 2-3 In some embodiments, an alkenyl group has 2 to 5 carbon atoms (“C 2-5 In some embodiments, an alkenyl group has 2 to 4 carbon atoms (“C 2-4 In some embodiments, an alkenyl group has 2 to 5 carbon atoms (“C 2-3 In some embodiments, an alkenyl group has 2 carbon atoms (“C 2 The one or more carbon-carbon double bonds may be internal (as in 2-butenyl) or terminal (as in 1-butenyl). 2–4 Examples of alkenyl groups include vinyl (C 2 ), 1-propenyl (C 3 ), 2-propenyl (C 3 ), 1-butenyl (C 4 ), 2-butenyl (C 4 ), butadiene (C 4 ) etc. 2–6 Examples of alkenyl groups include the above-mentioned C 2–4 Alkenyl and pentenyl (C 5 ), pentadienyl (C 5 ), hexenyl (C 6 ) etc. Other examples of alkenyl groups include heptenyl (C 7 ), octenyl (C 8 ), octatrienyl (C 8 ) etc. Unless otherwise specified, each instance of an alkenyl group is independently unsubstituted (an “unsubstituted alkenyl group”) or substituted (a “substituted alkenyl group”) with one or more substituents. In certain embodiments, an alkenyl group is an unsubstituted C 2–10 In certain embodiments, alkenyl is a substituted C 2–10 In alkenyl, the stereochemistry of an unspecified C=C double bond (e.g., -CH=CHCH 3 or ) can be in (E)- or (Z)-configuration.

[0029] The term "alkynyl" refers to a straight or branched chain hydrocarbon radical ("C 2–20 In some embodiments, an alkynyl group has 2 to 10 carbon atoms (“C 2–10 In some embodiments, an alkynyl group has 2 to 9 carbon atoms (“C 2-9 In some embodiments, an alkynyl group has 2 to 8 carbon atoms (“C 2-8 In some embodiments, an alkynyl group has 2 to 7 carbon atoms (“C 2-7 In some embodiments, an alkynyl group has 2 to 6 carbon atoms (“C 2-6 In some embodiments, an alkynyl group has 2 to 5 carbon atoms (“C 2-5 In some embodiments, an alkynyl group has 2 to 4 carbon atoms (“C 2-4 In some embodiments, an alkynyl group has 2 to 3 carbon atoms (“C 2-3 In some embodiments, an alkynyl group has 2 carbon atoms (“C 2 The one or more carbon-carbon triple bonds may be internal (as in 2-butynyl) or terminal (as in 1-butynyl). 2–4 Examples of alkynyl groups include, but are not limited to, ethynyl (C 2 ), 1-propynyl (C 3 ), 2-propynyl (C 3 ), 1-butynyl (C 4 ), 2-butynyl (C 4 ) etc. 2–6 Examples of alkenyl groups include the above-mentioned C 2–4 Alkynyl and pentynyl (C 5 ), hexynyl (C 6 ) and the like. Other examples of alkynyl groups include heptynyl (C 7 ), octynyl (C 8 ) etc. Unless otherwise specified, each instance of an alkynyl group is independently optionally substituted, i.e., unsubstituted (an “unsubstituted alkynyl group”) or substituted with one or more substituents (a “substituted alkynyl group”). In certain embodiments, an alkynyl group is an unsubstituted C 2–10 In certain embodiments, the alkynyl group is a substituted C 2–10 Alkynyl.

[0030] The term "carbocyclyl" or "carbocyclic" refers to a non-aromatic cyclic hydrocarbon radical having 3 to 10 ring carbon atoms ("C 3–10 In some embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms (“C 3–8In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms (“C 3-6 In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms (“C 3-6 In some embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms (“C 5-10 "Carbocyclyl"). Exemplary C 3–6 Carbocyclic groups include but are not limited to cyclopropyl (C 3 ), cyclopropenyl (C 3 ), cyclobutyl (C 4 ), cyclobutenyl (C 4 ), cyclopentyl (C 5 ), cyclopentenyl (C 5 ), cyclohexyl (C 6 ), cyclohexenyl (C 6 ), cyclohexadienyl (C 6 ) etc. Exemplary C 3–8 Carbocyclic groups include but are not limited to the above C 3–6 Carbocyclic and cycloheptyl (C 7 ), cycloheptenyl (C 7 ), cycloheptadienyl (C 7 ), cycloheptatrienyl (C 7 ), cyclooctyl (C 8 ), cyclooctenyl (C 8 ), bicyclo[2.2.1]heptyl (C 7 ), bicyclo[2.2.2]octyl (C 8 ) etc. Exemplary C 3–10 Carbocyclic groups include but are not limited to the above C 3–8 Carbocyclic and cyclononyl (C 9 ), cyclononenyl (C 9 ), cyclodecyl (C 10 ), cyclodecenyl (C 10 ), octahydro-1H-indenyl (C 9 ), decahydronaphthyl (C 10 ), spiro[4.5]decyl (C 10) and the like. As described in the foregoing examples, in certain embodiments, the carbocyclyl is monocyclic ("monocyclic carbocyclyl") or contains a fused, bridged or spirocyclic system, such as a bicyclic system ("bicyclic carbocyclyl") and can be saturated or can be partially unsaturated. "Carbocyclyl" also includes a ring system in which the carbocyclic ring as defined above is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the carbocyclic ring, and in such cases, the number of carbons continues to specify the number of carbons in the carbocyclic ring system. Unless otherwise indicated, each example of a carbocyclyl is independently optionally substituted, i.e., unsubstituted ("unsubstituted carbocyclyl") or substituted ("substituted carbocyclyl") by one or more substituents. In certain embodiments, a carbocyclyl is an unsubstituted C 3–10 In certain embodiments, the carbocyclyl group is a substituted C 3–10 Carbocyclic group.

[0031] In some embodiments, a "carbocyclyl" is a monocyclic saturated carbocyclyl ("C 3–10 In some embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms (“C 3–8 In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms (“C 3-6 In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C 5-6 In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms (“C 5-10 (cycloalkyl) 5–6 Examples of cycloalkyl groups include cyclopentyl (C 5 ) and cyclohexyl (C 5 ). C 3–6 Examples of the cycloalkyl group include the above-mentioned C 5–6 Cycloalkyl and cyclopropyl (C 3 ) and cyclobutyl (C 4 ). C 3–8 Examples of the cycloalkyl group include the above-mentioned C 3–6 Cycloalkyl and cycloheptyl (C 7 ) and cyclooctyl (C 8 Unless otherwise specified, each instance of a cycloalkyl group is independently unsubstituted (an “unsubstituted cycloalkyl group”) or substituted (a “substituted cycloalkyl group”) with one or more substituents. In certain embodiments, a cycloalkyl group is an unsubstituted C 3–10 In certain embodiments, the cycloalkyl group is a substituted C 3–10 Cycloalkyl. In certain embodiments, the carbocyclyl contains 0, 1, or 2 C=C double bonds in the carbocyclic ring system, as valency permits.

[0032] The term "heterocyclyl" or "heterocycle" refers to a radical of a 3- to 10-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from the group consisting of nitrogen, oxygen, sulfur, boron, phosphorus and silicon ("3- to 10-membered heterocyclyl"). In heterocyclyls containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom, where valence permits. A heterocyclyl may be a monocyclic ("monocyclic heterocyclyl") or a fused, bridged or spiro ring system, such as a bicyclic ring system ("bicyclic heterocyclyl"), and may be saturated or may be partially unsaturated. Heterocyclyl bicyclic ring systems may contain one or more heteroatoms in one or both rings. "Heterocyclyl" also includes ring systems in which a heterocycle as defined above is fused to one or more carbocyclyls, wherein the point of attachment is on the carbocyclyl or heterocycle, or a heterocycle as defined above is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocycle, and in such cases, the number of ring members continues to specify the number of ring members in the heterocycle system. Unless otherwise specified, each instance of a heterocyclyl is independently optionally substituted, i.e., unsubstituted ("unsubstituted heterocyclyl") or substituted ("substituted heterocyclyl") with one or more substituents. In certain embodiments, a heterocyclyl is an unsubstituted 3 to 10-membered heterocyclyl. In certain embodiments, a heterocyclyl is a substituted 3 to 10-membered heterocyclyl. In certain embodiments, a heterocyclyl is a substituted or unsubstituted 3 to 7-membered monocyclic heterocyclyl, wherein 1, 2, or 3 atoms in the heterocycle system are independently oxygen, nitrogen, or sulfur, as valence permits.

[0033] In some embodiments, the heterocyclyl group is a 5-10 membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from the group consisting of nitrogen, oxygen, sulfur, boron, phosphorus and silicon ("5-10 membered heterocyclyl"). In some embodiments, the heterocyclyl group is a 5-8 membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from the group consisting of nitrogen, oxygen and sulfur ("5-8 membered heterocyclyl"). In some embodiments, the heterocyclyl group is a 5-6 membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from the group consisting of nitrogen, oxygen and sulfur ("5-6 membered heterocyclyl"). In some embodiments, the 5-6 membered heterocyclyl group has 1 to 3 ring heteroatoms selected from nitrogen, oxygen and sulfur. In some embodiments, the 5-6 membered heterocyclyl group has 1 to 2 ring heteroatoms selected from nitrogen, oxygen and sulfur. In some embodiments, the 5-6 membered heterocyclyl group has one ring heteroatom selected from nitrogen, oxygen and sulfur.

[0034] Exemplary 3-membered heterocyclic groups containing one heteroatom include, but are not limited to, aziridine, oxirane, and thiirane. Exemplary 4-membered heterocyclic groups containing one heteroatom include, but are not limited to, azetidinyl, oxetane, and thiidine. Exemplary 5-membered heterocyclic groups containing one heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclic groups containing two heteroatoms include, but are not limited to, dioxolane, oxathiurea, dithiurea, and oxazolidin-2-one. Exemplary 5-membered heterocyclic groups containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic groups containing one heteroatom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, piperazinyl, morpholinyl, dithianyl, and dioxanyl. Exemplary 6-membered heterocyclic groups containing three heteroatoms include, but are not limited to, triazinyl. Exemplary 7-membered heterocyclic groups containing one heteroatom include, but are not limited to, azepanyl, oxepane, and thiepanyl. Exemplary 8-membered heterocyclic groups containing one heteroatom include, but are not limited to, azocanyl, oxepane, and thiecanyl. 6 Exemplary aryl ring-fused 5-membered heterocyclyl groups (also referred to herein as 5,6-bicyclic heterocycles) include, but are not limited to, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothiophenyl, benzoxazolinonyl, and the like. Exemplary 6-membered heterocyclic groups fused to an aryl ring (also referred to herein as 6,6-bicyclic heterocycles) include, but are not limited to, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, decahydroisoquinolinyl, octahydrochromenyl, octahydroisochromenyl, decahydronaphthyridinyl, decahydro-1,8-naphthyridinyl, octahydropyrrolo[3,2-b]pyrrole, indolinyl, phthalimidyl, naphthylimidyl, chromanyl, chromenyl, 1H-benzo[e][1,4]diazepinyl, 1,4,5,7-tetrahydropyrano[3,4-b]pyrrolyl, 5,6-dihydro-4H-furo[1,8-b]pyrrolyl, 1 ... 3,2-b]pyrrolyl, 6,7-dihydro-5H-furo[3,2-b]pyranyl, 5,7-dihydro-4H-thieno[2,3-c]pyranyl, 2,3-dihydro-1H-pyrrolo[2,3-b]pyridinyl, 2,3-dihydrofuro[2,3-b]pyridinyl, 4,5,6,7-tetrahydro-1H-pyrrolo[2,3-b]pyridinyl, 4,5,6,7-tetrahydrofuro[3,2-c]pyridinyl, 4,5,6,7-tetrahydrothieno[3,2-b]pyridinyl, 1,2,3,4-tetrahydro-1,6-naphthyridinyl, and the like.

[0035] The term "aryl" refers to a group having 6 to 14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system, a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10 or 14 π electrons shared in the cyclic array) ("C 6–14 In some embodiments, an aryl group has six ring carbon atoms (“C 6 In some embodiments, an aryl group has ten ring carbon atoms (“C 10 In some embodiments, an aryl group has fourteen ring carbon atoms (“C 14 "Aryl"; e.g., anthracenyl). "Aryl" also includes ring systems in which an aryl ring, as defined above, is fused to one or more carbocyclyl or heterocyclyl groups, wherein the radical or point of attachment is on the aryl ring, and in such cases, the number of carbon atoms continues to specify the number of carbon atoms in the aryl ring system. Unless otherwise specified, each instance of aryl is independently optionally substituted, i.e., unsubstituted ("unsubstituted aryl") or substituted ("substituted aryl") with one or more substituents. In certain embodiments, aryl is unsubstituted C 6–14 In certain embodiments, aryl is substituted C 6–14 Aryl.

[0036] The term "aralkyl" is a subset of "alkyl" and refers to an alkyl substituted with an aryl group, wherein the point of attachment is on the alkyl portion. In certain embodiments, the aralkyl is an optionally substituted benzyl. In certain embodiments, the aralkyl is a benzyl. In certain embodiments, the aralkyl is an optionally substituted phenethyl. In certain embodiments, the aralkyl is a phenethyl.

[0037] The term "heteroaryl" refers to a radical of a 5- to 10-membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 π electrons shared in a cyclic array) having ring carbon atoms and 1 to 4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from the group consisting of nitrogen, oxygen, and sulfur ("5- to 10-membered heteroaryl"). In heteroaryl groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom, where valence permits. Heteroaryl bicyclic ring systems may contain one or more heteroatoms in one or both rings. "Heteroaryl" includes ring systems in which a heteroaryl ring as defined above is fused to one or more carbocyclyl or heterocyclyl groups, wherein the point of attachment is on the heteroaryl ring, and in such cases, the number of ring members continues to specify the number of ring members in the heteroaryl ring system. "Heteroaryl" also includes ring systems in which a heteroaryl ring as defined above is fused to one or more aryl groups, wherein the point of attachment is on the aryl or heteroaryl ring, and in such cases, the number of ring members specifies the number of ring members in the fused (aryl / heteroaryl) ring system. For bicyclic heteroaryls in which one ring does not contain heteroatoms (e.g., indolyl, quinolyl, carbazolyl, etc.), the point of attachment can be on either ring, i.e., a ring with heteroatoms (e.g., 2-indolyl) or a ring without heteroatoms (e.g., 5-indolyl). In certain embodiments, heteroaryl is a substituted or unsubstituted 5- or 6-membered monocyclic heteroaryl, wherein 1, 2, 3, or 4 atoms in the heteroaryl ring system are independently oxygen, nitrogen, or sulfur. In certain embodiments, heteroaryl is a substituted or unsubstituted 9- or 10-membered bicyclic heteroaryl, wherein 1, 2, 3, or 4 atoms in the heteroaryl ring system are independently oxygen, nitrogen, or sulfur.

[0038] In some embodiments, heteroaryl is a 5 to 10 membered aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from the group consisting of nitrogen, oxygen and sulfur ("5 to 10 membered heteroaryl"). In some embodiments, heteroaryl is a 5 to 8 membered aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from the group consisting of nitrogen, oxygen and sulfur ("5 to 8 membered heteroaryl"). In some embodiments, heteroaryl is a 5 to 6 membered aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from the group consisting of nitrogen, oxygen and sulfur ("5 to 6 membered heteroaryl"). In some embodiments, 5 to 6 membered heteroaryl has 1 to 3 ring heteroatoms selected from nitrogen, oxygen and sulfur. In some embodiments, 5 to 6 membered heteroaryl has 1 to 2 ring heteroatoms selected from nitrogen, oxygen and sulfur. In some embodiments, the 5- to 6-membered heteroaryl group has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur. Unless otherwise indicated, each instance of a heteroaryl group is independently optionally substituted, i.e., unsubstituted ("unsubstituted heteroaryl") or substituted ("substituted heteroaryl") with one or more substituents. In certain embodiments, a heteroaryl group is an unsubstituted 5- to 14-membered heteroaryl group. In certain embodiments, a heteroaryl group is a substituted 5- to 14-membered heteroaryl group.

[0039] Exemplary 5-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyrrolyl, furanyl, and thienyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing one heteroatom include, but are not limited to, azepanyl, oxepane, and thiacycloheptane. Exemplary 5,6-bicyclic heteroaryls include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothienyl, isobenzothienyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryls include, but are not limited to, naphthyridinyl, pteridinyl, quinolyl, isoquinolyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. Exemplary tricyclic heteroaryls include phenanthridinyl, dibenzofuranyl, carbazolyl, acridinyl, phenothiazinyl, phenoxazinyl, and phenazinyl.

[0040] The term "heteroaralkyl" is a subset of "alkyl" and refers to an alkyl group substituted with a heteroaryl group, wherein the bond is on the alkyl portion.

[0041] The term "unsaturated" or "partially unsaturated" refers to a moiety containing at least one double or triple bond. "Partially unsaturated" ring systems are also intended to encompass rings with multiple sites of unsaturation, but are not intended to contain aromatic groups (e.g., aryl or heteroaryl) as defined herein. Likewise, "saturated" refers to groups that do not contain double or triple bonds, i.e., contain all single bonds.

[0042] Alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl groups as divalent bridging groups are further indicated using the suffix -ene, for example, alkylene, alkenylene, alkynylene, carbocyclylene, heterocyclylene, arylene and heteroarylene.

[0043] Unless expressly stated otherwise, a group is optionally substituted. The term "optionally substituted" means substituted or unsubstituted.

[0044] In certain embodiments, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl are optionally substituted (e.g., "substituted" or "unsubstituted" alkyl, "substituted" or "unsubstituted" alkenyl, "substituted" or "unsubstituted" alkynyl, heteroalkyl, "substituted" or "unsubstituted" heteroalkenyl, "substituted" or "unsubstituted" heteroalkynyl, "substituted" or "unsubstituted" carbocyclyl, "substituted" or "unsubstituted" heterocyclyl, "substituted" or "unsubstituted" aryl, or "substituted" or "unsubstituted" heteroaryl). In general, the term "substituted" refers to the replacement of at least one hydrogen present on a group with a permissible substituent, for example, a substituent that, when substituted, produces a stable compound, for example, a compound that does not spontaneously undergo transformation (such as by rearrangement, cyclization, elimination or other reaction). Unless otherwise specified, a "substituted" group has a substituent at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituent is the same or different at each position. The term "substituted" is contemplated to include substitution with all permissible substituents of organic compounds, and includes any substituent described herein that results in the formation of stable compounds. The present disclosure contemplates any and all such combinations to obtain stable compounds. For purposes of the present disclosure, heteroatoms such as nitrogen may have hydrogen substituents and / or any suitable substituents described herein that satisfy the valence of the heteroatoms and result in the formation of a stable moiety.

[0045] Exemplary carbon atom substituents include, but are not limited to, halogen, -CN, -NO 2 、-N 3 、-SO 2H、-SO 3 H、-OH、-OR aa 、-ON(R bb ) 2 、-N(R bb ) 2 、-N(R bb ) 3 + X - 、-N(OR cc )R bb 、-SH、-SR aa 、-SSR cc 、-C(=O)R aa 、-CO 2 H、-CHO、-C(OR cc ) 2 、-CO 2 R aa 、-OC(=O)R aa 、-OCO 2 R aa 、-C(=O)N(R bb ) 2 、-OC(=O)N(R bb ) 2 、-NR bb C(=O)R aa 、-NR bb CO 2 R aa 、-NR bb C(=O)N(R bb ) 2 、-C(=NR bb )R aa 、-C(=NR bb )OR aa 、-OC(=NRbb)R aa 、-OC(=NR bb )OR aa 、-C(=NR bb )N(R bb ) 2 、-OC(=NR bb )N(R bb ) 2 、-NR bb C(=NR bb )N(R bb ) 2 、-C(=O)NR bb SO 2 R aa 、-NR bb SO 2 Raa 、-SO 2 N(R bb ) 2 、-SO 2 R aa 、-SO 2 OR aa 、-OSO 2 R aa 、-S(=O)R aa 、-OS(=O)R aa 、-Si(R aa ) 3 、-OSi(R aa ) 3 -C(=S)N(R bb ) 2 、-C(=O)SR aa 、-C(=S)SR aa 、-SC(=S)SR aa 、-SC(=O)SR aa 、-OC(=O)SR aa 、-SC(=O)OR aa 、-SC(=O)R aa 、-P(=O)(R aa ) 2 、-P(=O)(OR cc ) 2 、-OP(=O)(R aa ) 2 、-OP(=O)(OR cc ) 2 、-P(=O)(N(R bb ) 2 ) 2 、-OP(=O)(N(R bb ) 2 ) 2 、-NR bb P(=O)(R aa ) 2 、-NR bb P(=O)(OR cc ) 2 、-NR bb P(=O)(N(R bb ) 2 ) 2 、-P(R cc ) 2 、-P(OR cc ) 2 、-P(R cc) 3 + X -、-P(OR cc ) 3 + X - 、-P(R cc ) 4 、-P(OR cc ) 4 、-OP(R cc ) 2 、-OP(R cc ) 3 + X - 、-OP(OR cc ) 2 、-OP(OR cc ) 3 + X - 、-OP(R cc ) 4 、-OP(OR cc ) 4 ,-B(R aa ) 2 、-B(OR cc ) 2 , -BR aa (OR cc ), C 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, hetero C 1-10 Alkyl, hetero C 2-10 Alkenyl, hetero C 2-10 Alkynyl, C 3-10 Carbocyclic group, 3 to 14 membered heterocyclic group, C 6-14 aryl and 5- to 14-membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd Group substitution; wherein X - is the counter ion;

[0046] Or the two geminal hydrogens on the carbon atom are replaced by groups =O, =S, =NN(R bb ) 2 , =NNR bb C(=O)R aa , =NNR bb C(=O)OR aa , =NNR bb S(=O) 2 R aa , =NR bb or = NOR cc Replacement;

[0047] R aa Each instance of is independently selected from C 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, hetero C 1-10 Alkyl, hetero C 2-10 Alkenyl, hetero C 2-10 Alkynyl, C 3-10 Carbocyclic group, 3 to 14 membered heterocyclic group, C 6-14 aryl and 5- to 14-membered heteroaryl, or two R aa The groups are connected to form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd group substitution;

[0048] R bb Each instance of is independently selected from hydrogen, -OH, -OR aa 、-N(R cc ) 2 、-CN、-C(=O)R aa 、-C(=O)N(R cc ) 2 , -CO 2 R aa 、-SO 2 R aa 、-C(=NR cc )OR aa 、-C(=NR cc )N(R cc) 2 、-SO 2 N(R cc ) 2 、-SO 2 R cc 、-SO 2 OR cc 、-SOR aa 、-C(=S)N(R cc ) 2 、-C(=O)SR cc 、-C(=S)SR cc ,-P(=O)(R aa ) 2 、-P(=O)(OR cc ) 2 、-P(=O)(N(R cc ) 2 ) 2 , C1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, hetero C 1-10 Alkyl, hetero C 2-10 Alkenyl, hetero C 2-10 Alkynyl, C 3-10 Carbocyclic group, 3 to 14 membered heterocyclic group, C 6-14 aryl and 5- to 14-membered heteroaryl, or two R bb The groups are connected to form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd Group substitution; wherein X - is the counter ion;

[0049] R cc Each instance of is independently selected from hydrogen, C 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, hetero C 1-10 Alkyl, hetero C 2-10 Alkenyl, hetero C 2-10 Alkynyl, C 3-10 Carbocyclic group, 3 to 14 membered heterocyclic group, C 6-14 aryl and 5- to 14-membered heteroaryl, or two R cc The groups are connected to form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd group substitution;

[0050] R dd Each instance of is independently selected from halogen, -CN, -NO 2 、-N 3 、-SO 2 H, -SO 3 H, -OH, -OR ee 、-ON(R ff ) 2 、-N(R ff ) 2 、-N(R ff ) 3 + X - 、-N(OR ee )R ff , -SH, -SR ee 、-SSR ee、-C(=O)R ee 、-CO 2 H、-CO 2 R ee 、-OC(=O)R ee 、-OCO 2 R ee 、-C(=O)N(R ff ) 2 、-OC(=O)N(R ff ) 2 、-NR ff C(=O)R ee 、-NR ff CO 2 R ee 、-NR ff C(=O)N(R ff ) 2 、-C(=NR ff )OR ee 、-OC(=NR ff )R ee 、-OC(=NR ff )OR ee 、-C(=NR ff )N(R ff ) 2 、-OC(=NR ff )N(R ff ) 2 、-NR ff C(=NR ff )N(R ff) 2 、-NR ff SO 2 R ee 、-SO2N(R ff ) 2 、-SO 2 R ee 、-SO 2 OR ee 、-OSO 2 R ee 、-S(=O)R ee 、-Si(R ee ) 3 、-OSi(R ee ) 3 、-C(=S)N(R ff ) 2 、-C(=O)SR ee 、-C(=S)SR ee 、-SC(=S)SR ee 、-P(=O)(OR ee) 2 ,-P(=O)(R ee ) 2 ,-OP(=O)(R ee ) 2 ,-OP(=O)(OR ee ) 2 , C 1-6 Alkyl, C 1-6 Perhaloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, hetero C 1-6 Alkyl, hetero C 2-6 Alkenyl, hetero C 2-6 Alkynyl, C 3-10 Carbocyclic group, 3 to 10 membered heterocyclic group, C 6-10 aryl and 5- to 10-membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R gg Group substitution, or two geminal R dd Substituents may be linked to form =O or =S; where X - is the counter ion;

[0051] R ee Each instance of is independently selected from C 1-6 Alkyl, C 1-6 Perhaloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, hetero C 1-6 Alkyl, hetero C 2-6 Alkenyl, hetero C 2-6 Alkynyl, C 3-10 Carbocyclic group, C 6-10 aryl, 3- to 10-membered heterocyclyl, and 3- to 10-membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted by 0, 1, 2, 3, 4, or 5 R gg Group substitution;

[0052] R ff Each instance of is independently selected from hydrogen, C 1-6 Alkyl, C 1-6 Perhaloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, hetero C 1-6 Alkyl, hetero C 2-6 Alkenyl, hetero C 2-6 Alkynyl, C 3-10 Carbocyclic group, 3 to 10 membered heterocyclic group, C 6-10 aryl and 5- to 10-membered heteroaryl, or two R ffThe groups are connected to form a 3- to 10-membered heterocyclyl or 5- to 10-membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R gg Group substitution; and

[0053] R gg Each instance of is independently halogen, -CN, -NO 2 、-N 3 、-SO 2 H, -SO 3 H, -OH, -OC 1-6 Alkyl, -ON(C 1-6 alkyl) 2 、-N(C 1-6 alkyl) 2 、-N(C 1-6 alkyl) 3 + X - 、-NH(C 1-6 alkyl) 2 + X - 、-NH 2 (C 1-6 alkyl) + X - 、-NH 3 + X - 、-N(OC 1-6 Alkyl)(C 1-6 Alkyl), -N(OH)(C 1-6 Alkyl), -NH(OH), -SH, -SC 1-6 Alkyl, -SS(C 1-6 alkyl), -C(=O)(C 1-6 Alkyl), -CO 2 H, -CO 2 (C 1-6 alkyl), -OC(=O)(C 1-6 Alkyl), -OCO 2 (C 1-6 alkyl), -C(=O)NH 2 、-C(=O)N(C 1-6 alkyl) 2 、-OC(=O)NH(C 1-6 alkyl), -NHC(=O)(C 1-6 Alkyl), -N(C 1-6 alkyl)C(=O)(C 1-6 Alkyl), -NHCO 2 (C 1-6alkyl), -NHC(=O)N(C 1-6 alkyl) 2 、-NHC(=O)NH(C 1-6 alkyl), -NHC(=O)NH 2 、-C(=NH)O(C 1-6 alkyl), -OC(=NH)(C 1-6 alkyl), -OC(=NH)OC 1-6 Alkyl, -C(=NH)N(C 1-6 alkyl) 2 、-C(=NH)NH(C 1-6 alkyl), -C(=NH)NH 2 、-OC(=NH)N(C 1-6 alkyl) 2 、-OC(NH)NH(C 1-6 Alkyl), -OC(NH)NH 2 、-NHC(NH)N(C 1-6 alkyl) 2 、-NHC(=NH)NH 2 、-NHSO 2 (C 1-6 Alkyl), -SO 2 N(C 1-6 alkyl) 2 、-SO 2 NH(C 1-6 Alkyl), -SO 2 NH 2 、-SO 2 C 1-6 Alkyl, -SO 2 OC 1-6 Alkyl, -OSO 2 C 1-6 Alkyl, -SOC 1-6 Alkyl, -Si(C 1-6 alkyl) 3 、-OSi(C 1-6 alkyl) 3 -C(=S)N(C 1-6 alkyl) 2 、C(=S)NH(C 1-6 alkyl), C(=S)NH 2 、-C(=O)S(C 1-6 Alkyl), -C(=S)SC 1-6 Alkyl, -SC(=S)SC 1-6 Alkyl, -P(=O)(OC 1-6 alkyl) 2 ,-P(=O)(C 1-6 alkyl)2 ,-OP(=O)(C 1-6 alkyl) 2 ,-OP(=O)(OC 1-6 alkyl) 2 , C 1-6 Alkyl, C 1-6 Perhaloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, hetero C 1-6 Alkyl, hetero C 2-6 Alkenyl, hetero C 2-6 Alkynyl, C 3-10 Carbocyclic group, C 6-10 aryl, 3- to 10-membered heterocyclyl or 5- to 10-membered heteroaryl; or two geminal R gg Substituents may be linked to form =O or =S; where X - It is a counter ion.

[0054] A "counterion" or "anionic counterion" is a negatively charged group that associates with a positively charged group to maintain electrical neutrality. Anionic counterions can be monovalent (i.e., contain one formal negative charge). Anionic counterions can also be multivalent (i.e., contain more than one formal negative charge), such as divalent or trivalent. Exemplary counterions include halides (e.g., F – , Cl – Br – ,I – ), NO 3 – , ClO 4 – OH – , H 2 PO 4 – , HCO 3 - , HSO 4 – , sulfonate ions (e.g., methanesulfonate, trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate, 10-camphorsulfonate, naphthalene-2-sulfonate, naphthalene-1-sulfonic acid-5-sulfonate, ethane-1-sulfonic acid-2-sulfonate, etc.), carboxylate ions (e.g., acetate, propionate, benzoate, glycerate, lactate, tartrate, glycolate, gluconate, etc.), BF 4 - PF 4 – PF 6 – , AsF 6 – , SbF 6 – , B[3,5-(CF3 ) 2 C 6 H 3 ] 4 ] – , B(C 6 F 5 ) 4 - BPh 4 – 、Al(OC(CF 3 ) 3 ) 4 – and carborane anions (e.g., CB 11 H 12 – or (HCB 11 Me 5 Br 6 ) – ). Exemplary counterions that may be multivalent include CO 3 2- , HPO 4 2- ,PO 4 3- , B 4 O 7 2- 、SO 4 2- , S 2 O 3 2- , carboxylate anions (e.g., tartrate, citrate, fumarate, maleate, malate, malonate, gluconate, succinate, glutarate, adipate, pimelate, suberate, azelaate, sebacate, salicylate, phthalate, aspartate, glutamate, etc.), and carborane.

[0055] "Halo" or "halogen" refers to fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br), or iodine (iodo, -I).

[0056] The term "acyl" refers to an acyl group having the general formula -C(=O)R X1 、–C(=O)OR X1 、–C(=O)–O–C(=O)R X1 、–C(=O)SR X1 、–C(=O)N(R X1) 2 , –C(=S)R X1 , –C(=S)N(R X1 ) 2 , –C(=S)S(R X1),–C(=NR X1 )R X1 , –C(=NR X1 )OR X1 , –C(=NR X1 )SR X1 or –C(=NR X1 )N(R X1 ) 2 A group in which each R X1 are independently hydrogen; halogen; substituted or unsubstituted hydroxy; substituted or unsubstituted thiol; substituted or unsubstituted amino; substituted or unsubstituted acyl; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched alkyl; cyclic or acyclic, substituted or unsubstituted, branched or unbranched alkenyl; substituted or unsubstituted alkynyl; substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, aliphaticoxy, heteroaliphaticoxy, alkoxy, heteroalkoxy, aryloxy, heteroaryloxy, aliphaticthiooxy, heteroaliphaticthiooxy, alkylthiooxy, heteroalkylthiooxy, arylthiooxy, heteroarylthiooxy, mono- or di-aliphatic amino, mono- or di-heteroaliphatic amino, mono- or di-alkylamino, mono- or di-heteroalkylamino, mono- or di-arylamino, or mono- or di-heteroarylamino; or two R X1 The groups together form a 5- to 6-membered heterocyclic ring. Exemplary acyl groups include aldehydes (-CHO), carboxylic acids (-CO 2 H), ketones, acyl halides, esters, amides, imines, carbonates, carbamates and ureas. Acyl substituents include, but are not limited to, any substituent described herein that results in the formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, oxo, imino, thio, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halogen, aliphatic amino, heteroaliphatic amino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphatic oxy, heteroaliphatic oxy, alkoxy, heteroalkoxy, aryloxy, heteroaryloxy, aliphatic thiooxy, heteroaliphatic thiooxy, alkylthiooxy, heteroalkylthiooxy, arylthiooxy, heteroarylthiooxy, acyloxy, etc., each of which may be further substituted or unsubstituted).

[0057] "Alkoxy" or "alkoxyl" refers to a radical of the formula: -O-alkyl.

[0058] Where valence permits, the nitrogen atoms may be substituted or unsubstituted and include primary, secondary, tertiary, and quaternary nitrogen atoms. Exemplary nitrogen atom substituents include, but are not limited to, hydrogen, -OH, -ORaa 、-N(R cc ) 2 、-CN、-C(=O)R aa 、-C(=O)N(R cc ) 2 , -CO 2 R aa 、-SO 2 R aa 、-C(=NR bb )R aa 、-C(=NR cc )OR aa 、-C(=NR cc )N(R cc ) 2 、-SO 2 N(R cc ) 2 、-SO 2 R cc 、-SO 2 OR cc 、-SOR aa 、-C(=S)N(R cc ) 2 、-C(=O)SR cc 、-C(=S)SR cc 、-P(=O)(OR cc ) 2 ,-P(=O)(R aa ) 2 、-P(=O)(N(R cc ) 2 ) 2 , C 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, hetero C 1-10 Alkyl, hetero C 2-10 Alkenyl, hetero C 2-10 Alkynyl, C 3-10 Carbocyclic group, 3 to 14 membered heterocyclic group, C 6-14 Aryl and 5- to 14-membered heteroaryl, or two R groups attached to the N atom cc The groups are connected to form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd group substituted, and wherein R aa , R bb , R cc and R dd As defined above.

[0059] In certain embodiments, each nitrogen atom substituent is independently substituted (eg, by one or more halogens) or unsubstituted C 1-6 Alkyl, -C(=O)R aa , -CO 2 R aa 、-C(=O)N(R bb ) 2 In certain embodiments, each nitrogen atom substituent is independently substituted (e.g., substituted with one or more halogens) or unsubstituted C 1-10 Alkyl, -C(=O)R aa , -CO 2 R aa 、-C(=O)N(R bb ) 2 , or a nitrogen protecting group, where R aa is hydrogen, is substituted (e.g., by one or more halogens) or is unsubstituted C 1-10 alkyl, or an oxygen protecting group when attached to an oxygen atom; and each R bb are independently hydrogen, substituted (e.g., substituted with one or more halogens) or unsubstituted C 1-10 In certain embodiments, each nitrogen atom substituent is independently substituted (e.g., substituted with one or more halogens) or unsubstituted C 1-6 Alkyl or nitrogen protecting group.

[0060] In certain embodiments, the substituent present on the nitrogen atom is a nitrogen protecting group (also known as an amino protecting group). Nitrogen protecting groups include but are not limited to -OH, -OR aa , –N(R cc ) 2 , –C(=O)R aa 、–C(=O)N(R cc ) 2 , –CO 2 R aa ,–SO 2 R aa , –C(=NR cc )R aa , –C(=NR cc )OR aa , –C(=NR cc )N(R cc ) 2 ,–SO 2 N(R cc ) 2 ,–SO 2 R cc ,–SO2 OR cc , –SOR aa , –C(=S)N(R cc ) 2 、–C(=O)SR cc , –C(=S)SR cc , C 1–10 Alkyl (e.g., aralkyl, heteroaralkyl), C 2–10 Alkenyl, C 2–10 Alkynyl, C 3–10 Carbocyclic group, 3 to 14 membered heterocyclic group, C 6-14 aryl and 5- to 14-membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd group substituted, and wherein R aa , R bb , R cc and R dd As defined above. Nitrogen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3rd ed., John Wiley & Sons, 1999, which is incorporated herein by reference.

[0061] For example, a nitrogen protecting group such as an amide group (eg, -C(=O)R aa ) include, but are not limited to, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropionamide, picolinamide, 3-pyridylformamide, N-benzoylphenylalanyl derivatives, benzamide, p-phenylbenzamide, o-nitrobenzylacetamide, o-nitrophenoxyacetamide, acetoacetamide, (N′-dithiobenzyloxyamido)acetamide, 3-(p-hydroxyphenyl)propionamide, 3-(o-nitrophenoxy)propionamide, 2-methyl-2-(o-phenylphenoxy)propionamide, 2-methyl-2-(o-phenylazophenoxy)propionamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinnamamide, N-acetylmethionine derivatives, o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.

[0062] Nitrogen protecting groups, such as carbamate groups (e.g., -C(=O)OR aa) include but are not limited to methyl carbamate, ethyl carbamate, 9-fluorenylmethylcarbamate (Fmoc), 9-(2-sulfo)fluorenylmethylcarbamate, 9-(2,7-dibromo)fluoroenylmethylcarbamate, 2,7-di-tert-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthanone)]methyl carbamate (DBD-Tmoc), 4-methoxybenzylcarbamate (Phenoc), 2,2,2-trichloroethylcarbamate (Troc), 2-trimethylsilylethylcarbamate (Teoc), 2-phenylethylcarbamate (hZ), 1-(1-adamantyl)-1-methylethylcarbamate (Adp oc), 1,1-dimethyl-2-haloethylcarbamate, 1,1-dimethyl-2,2-dibromoethylcarbamate (DB–t–BOC), 1,1-dimethyl-2,2,2-trichloroethylcarbamate (TCBOC), 1-methyl-1-(4-biphenylyl)ethylcarbamate (Bpoc), 1-(3,5-di-tert-butylphenyl)-1-methylethylcarbamate (t–Bumeoc), 2-(2′- and 4′-pyridyl)ethylcarbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamido)ethylcarbamate, tert-butyl carbamate (BOC), 1-adamantylcarbamate (Adoc), vinylcarbamate (Vo c), allyl carbamate (Alloc), 1-isopropyl allyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyl dithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p-nitrobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate, 2-(p-toluenesulfonyl)ethylcarbamate, [2-(1,3-dithianyl)]methylcarbamate (Dmoc), 4-methylphenylthiocarbamate (Mtpc), 2,4-dimethylphenylthiocarbamate (Bmpc), 2-phosphonooxyethylcarbamate (Peoc), 2-triphenylphosphonimide isopropylcarbamate (Ppoc), 1,1-dimethyl-2-cyanoethylcarbamate, m-chloro-p-acyloxybenzylcarbamate, p-(dihydroxyboryl)benzylcarbamate, 5-benzisoxazolylmethylcarbamate, 2-(trifluoromethyl)-6-chromononylmethylcarbamate (Tcroc), m-nitrophenylcarbamate, 3,5-dimethoxybenzylcarbamate, o-nitrobenzylcarbamate, 3,4-dimethoxy-6-nitrobenzylcarbamate, phenyl (o-nitrophenyl) methylcarbamate, tert-amyl carbamate, S-benzylthiocarbamate, p-cyanophenylcarbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentyl carbamate, cyclopropyl methylcarbamate, p-decyloxybenzylcarbamate, 2,2-dimethoxyvinylcarbamate, o-(N,N-dimethylformamido)benzylcarbamate, 1,1-dimethyl-3-(N,N-dimethylformamido)propylcarbamate, 1,1-dimethylpropynylcarbamate, di(2-pyridyl)methylcarbamate, 2-furylmethylcarbamate, 2-iodoethylamino carbamate, isoborynyl carbamate, isobutyl carbamate, isonicotinoyl carbamate, p-(p'-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate, 1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate, 1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate, 2,4,6-tri-tert-butylphenyl carbamate, 4-(trimethylammonium)benzyl carbamate and 2,4,6-trimethylbenzyl carbamate.

[0063] Nitrogen protecting groups, such as sulfonamide groups (e.g., -S(=O)2R aa ) include but are not limited to toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms), β-trimethylsilylethylsulfonamide (SES), 9-anthracenesulfonamide, 4-(4′,8′-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide and benzoylsulfonamide.

[0064] Other nitrogen protecting groups include, but are not limited to, phenothiazinyl-(10)-acyl derivatives, N′-toluenesulfonylaminoacyl derivatives, N′-phenylaminothioacyl derivatives, N-benzoylphenylalanyl derivatives, N-acetylmethionine derivatives, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiosuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilazolidine adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexane ... ketones, 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexane-2-ones, 1-substituted 3,5-dinitro-4-pyridones, N-methylamine, N-allylamine, N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl)amine, quaternary ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzocycloheptylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenamine (PhF), N-2,7 -Dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm), N-2-pyridylamino N′-oxide, N-1,1-dimethylthiomethyleneamine, N-benzylideneamine, N-p-methoxybenzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)benzyltrimethyl]methyleneamine, N-(N′,N′-dimethylaminomethylene)amine, N,N′-isopropylidenediamine, N-p-nitrobenzylideneamine, N-salicylideneamine, N-5-chlorosalicylideneamine, N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexylamine, N-(5,5-dimethyl-3-oxo-1-cyclohexene In some embodiments, the nitrogen protecting group and two examples of the nitrogen atom to which the nitrogen protecting group is attached are N,N'-isopropylamine.

[0065] In certain embodiments, at least one nitrogen protecting group is Bn, Boc, Cbz, Fmoc, trifluoroacetyl, triphenylmethyl, acetyl, or Ts.

[0066] In certain embodiments, each oxygen atom substituent is independently substituted (eg, with one or more halogens) or unsubstituted C 1-10 Alkyl, -C(=O)R aa , -CO 2 R aa 、-C(=O)N(R bb ) 2 In certain embodiments, each oxygen atom substituent is independently substituted (e.g., substituted with one or more halogens) or unsubstituted C 1-6 Alkyl, -C(=O)R aa , -CO 2 R aa 、-C(=O)N(R bb ) 2 or an oxygen protecting group, where R aa is hydrogen, is substituted (e.g., by one or more halogens) or is unsubstituted C 1-10 alkyl, or, when attached to an oxygen atom, an oxygen protecting group; and each R bb is independently hydrogen, substituted (e.g., substituted with one or more halogens) or unsubstituted C 1-10 In certain embodiments, each oxygen atom substituent is independently substituted (e.g., substituted with one or more halogens) or unsubstituted C 1-6 Alkyl or oxygen protecting groups.

[0067] In certain embodiments, the substituent present on the oxygen atom is an oxygen protecting group (also referred to herein as a "hydroxy protecting group"). Oxygen protecting groups include but are not limited to: -R aa 、-N(R bb ) 2 、-C(=O)SR aa 、-C(=O)R aa , -CO 2 R aa 、-C(=O)N(R bb ) 2 、-C(=NR bb )R aa 、-C(=NR bb )OR aa 、-C(=NR bb )N(R bb ) 2 、-S(=O)R aa 、-SO 2 Raa 、-Si(R aa ) 3 、-P(R cc ) 2 、-P(R cc ) 3 + X - 、-P(OR cc ) 2 、-P(OR cc ) 3 + X - ,-P(=O)(R aa ) 2 、-P(=O)(OR cc ) 2 and -P(=O)(N(R bb ) 2 ) 2 , where X - , R aa , R bb and R cc As defined herein. Oxygen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, TW Green and PGM Woods, 3rd ed., John Wiley & Sons, 1999, which is incorporated herein by reference.

[0068] Exemplary oxygen protecting groups include, but are not limited to, methyl, methoxymethyl (MOM), methylthiomethyl (MTM), tert-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), tert-butoxymethyl, 4-pentenyloxymethyl (POM), silyloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyrrolidone, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl (CTMP), 1,4-dioxane-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methylbenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl 1-Benzyl-1-yloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylseleno)ethyl, tert-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-pyridylmethyl, 4-pyridylmethyl, 3-methyl-2-pyridylmethyl N-oxide, diphenylmethyl, p,p'-dinitrodiphenylmethyl, 5-dibenzosuberyl, triphenylmethyl, α-naphthyldiphenylmethyl, p-methyl Oxyphenylbenzhydryl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4'-bromobenzoylmethoxyphenyl)benzhydryl, 4,4',4"-tri(4,5-dichlorophthalimidophenyl)methyl, 4,4',4"-tri(acetylpropoxyphenyl)methyl, 4,4',4"-tri(benzoyloxyphenyl)methyl, 3-(imidazol-1-yl)bis(4',4"-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodisulfide-2-yl, S,S-dioxybenzisothiazolyl, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethyltert-hexylsilyl, tert-butyldimethylsilyl (TBDMS), tert-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), tert-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxy Acetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (levulinic acid ester), 4,4-(ethylenedithio)pentanoate (levulinic acid dithioacetal), pivalate, adamantium carboxylate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (2,4,6-trimethylbenzoate) (2,4,6-trimethylbenzoate (mesitoate)), alkyl carbonate methyl ester, 9-fluorenyl carbonate methyl ester (Fmoc), alkyl carbonate ethyl ester, alkyl carbonate 2,2,2-trichloroethyl (Troc), carbonyl 2-(Trimethylsilyl) ethyl carbonate (TMSEC), 2-(phenylsulfonyl) ethyl carbonate (Psec), 2-(triphenylphosphinimide) ethyl carbonate (Peoc), alkyl carbonate isobutyl ester, alkyl carbonate vinyl ester, alkyl carbonate allyl ester, alkyl carbonate p-nitrophenyl ester, alkyl carbonate benzyl ester, alkyl carbonate p-methoxybenzyl ester, alkyl carbonate 3,4-dimethoxybenzyl ester, alkyl carbonate o-nitrobenzyl ester, alkyl carbonate p-nitrobenzyl ester, alkyl thiocarbonate S-benzyl ester, 4-ethoxy-1-naphthyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylvalerate, o-(dibromo)benzyl carbonate 2-(Methyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, succinate, (E)-2-methyl-2-butenoate, o-(methoxyacyl)benzoate, α-naphthoate, nitrate, N,N,N′,N′-tetramethylphosphorodiamidate alkyl ester, N-phenylcarbamic acid alkyl ester, borate, dimethylthiophosphonyl, 2,4-Dinitrobenzene sulfenic acid alkyl esters, sulfates, methanesulfonates (mesylate), benzylsulfonates and toluenesulfonates (Ts).

[0069] In certain embodiments, at least one oxygen protecting group is silyl, TBDPS, TBDMS, TIPS, TES, TMS, MOM, THP, t-Bu, Bn, allyl, acetyl, pivaloyl, or benzoyl.

[0070] In certain embodiments, each sulfur atom substituent is independently substituted (eg, with one or more halogens) or unsubstituted C 1-10 Alkyl, -C(=O)R aa , -CO 2 R aa 、-C(=O)N(R bb ) 2 In certain embodiments, each sulfur atom substituent is independently substituted (e.g., substituted with one or more halogens) or unsubstituted C 1-10 Alkyl, -C(=O)R aa , -CO 2 R aa 、-C(=O)N(R bb ) 2 or a sulfur protecting group, wherein R aa is hydrogen, is substituted (e.g., by one or more halogens) or is unsubstituted C 1-10 alkyl, or an oxygen protecting group when attached to an oxygen atom; and each R bb is independently hydrogen, substituted (e.g., substituted with one or more halogens) or unsubstituted C 1-10 In certain embodiments, each sulfur atom substituent is independently substituted (e.g., substituted with one or more halogens) or unsubstituted C 1-6 Alkyl or sulfur protecting group.

[0071] In certain embodiments, the substituent present on the sulfur atom is a sulfur protecting group (also known as a "thiol protecting group").

[0072] In some embodiments, each sulfur protecting group is selected from the group consisting of: -R aa 、-N(R bb ) 2 、-C(=O)SR aa 、-C(=O)R aa , -CO 2 R aa 、-C(=O)N(R bb ) 2、-C(=NR bb )R aa 、-C(=NR bb )OR aa 、-C(=NR bb )N(R bb ) 2 、-S(=O)R aa 、-SO 2 R aa 、-Si(R aa ) 3 、-P(R cc ) 2 、-P(R cc ) 3 + X - 、-P(OR cc ) 2 、-P(OR cc ) 3 + X - ,-P(=O)(R aa ) 2 、-P(=O)(OR cc ) 2 and -P(=O)(N(R bb ) 2 ) 2 , where R aa , R bb and R cc As defined herein. Sulfur protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, TW Green and PGM Woods, 3rd ed., John Wiley & Sons, 1999, which is incorporated herein by reference.

[0073] As used herein, "leaving group" (LG) is a term understood in the art and refers to a molecular fragment that leaves with a pair of electrons in heterolytic bond cleavage, wherein the molecular fragment is an anion or a neutral molecule. As used herein, a leaving group can be an atom or group that can be displaced by a nucleophile. See, e.g., Smith, March Advanced Organic Chemistry, 6th edition (501-502). Exemplary leaving groups include, but are not limited to, halogens (e.g., chlorine, bromine, iodine) and activated substituted hydroxyls (e.g., -OC(=O)SR aa 、–OC(=O)R aa , –OCO 2 R aa 、–OC(=O)N(R bb ) 2 、–OC(=NR bb )R aa、–OC(=NR bb )OR aa 、–OC(=NR bb )N(R bb ) 2 、–OS(=O)R aa , –OSO 2 R aa ,–OP(R cc ) 2 ,–OP(R cc ) 3 ,–OP(=O)(R aa ) 2 ,–OP(=O)(OR cc ) 2 and –OP(=O)(NR bb ) 2 , where R aa , R bb and R cc As defined herein). Examples of suitable leaving groups include, but are not limited to, halogen (such as F, Cl, Br or I (iodine)), alkoxycarbonyloxy, aryloxycarbonyloxy, alkanesulfonyloxy, arenesulfonyloxy, alkyl-carbonyloxy (e.g., acetoxy), arylcarbonyloxy, aryloxy, methoxy, N, O-dimethylhydroxyamino, hexyloxy and haloformate. In some cases, the leaving group is a sulfonate, such as toluenesulfonate (toluenesulfonate, –OTs), mesylate (mesylate, –OMs), p-bromobenzenesulfonyloxy (brosylate, –OBs) or trifluoromethanesulfonate (triflate, –OTf). In some cases, the leaving group is brosylate, such as p-bromobenzenesulfonyloxy. In some cases, the leaving group is a nitrobenzenesulfonate, such as 2-nitrobenzenesulfonyloxy. In some embodiments, the leaving group is a sulfonate-containing group. In some embodiments, the leaving group is a tosylate group. The leaving group can also be a phosphine oxide (e.g., formed during the Mitsunobu reaction) or an internal leaving group such as an epoxide or a cyclic sulfate. Other non-limiting examples of leaving groups are water, amines, ammonia, alcohols, ether moieties, sulfur-containing moieties, thioether moieties, zinc halides, magnesium moieties, diazonium salts, and copper moieties.

[0074] The term "pharmaceutically acceptable salt" refers to salts that are suitable for contact with tissues of humans and lower animals without excessive toxicity, irritation, allergic reactions, etc., and have a reasonable benefit / risk ratio within the scope of reasonable medical judgment. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in "Journal of Pharmaceutical Sciences (J. Pharmaceutical Sciences)" 1977, 66, 1 to 19, which are incorporated herein by reference. Pharmaceutically acceptable salts of the compounds disclosed herein include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are amino salts formed with inorganic acids (such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid) or with organic acids (such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid) or by using other methods known in the art (such as ion exchange). Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N + (C 1–4 alkyl) 4 - Salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Where appropriate, other pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates and aryl sulfonates.

[0075] The term "solvate" refers to a form of a compound or a salt thereof that is associated with a solvent, typically by a solvolysis reaction. This physical association may include hydrogen bonds. Conventional solvents include water, methanol, ethanol, acetic acid, DMSO, THF, ether, and the like. Compound (I) may be prepared, for example, in a crystalline form and may be solvated. Suitable solvates include pharmaceutically acceptable solvates, and further include stoichiometric solvates and non-stoichiometric solvates. In some cases, the solvate will be able to separate, for example when one or more solvent molecules are incorporated into the crystal lattice of the crystalline solid. "Solvate" encompasses solution phases and separable solvates. Representative solvates include hydrates, ethanolates, and methanolates.

[0076] The term "stoichiometric solvate" refers to a solvate comprising a compound (e.g., a compound disclosed herein) and a solvent, wherein the solvent molecules are an integral part of the crystal lattice in which they interact strongly with the compound and each other. Removal of the solvent molecules will result in instability of the crystal network, which will subsequently collapse into an amorphous phase or recrystallize into a new crystalline form with a reduced solvent content.

[0077] The term "non-stoichiometric solvate" refers to a solvate comprising a compound (e.g., a compound disclosed herein) and a solvent, wherein the solvent content can vary without significant changes to the crystal structure. The amount of solvent in the crystal lattice depends only on the partial pressure of the solvent in the surrounding atmosphere. In the fully solvated state, a non-stoichiometric solvate can, but does not necessarily have to, show an integer molar ratio of solvent to compound. During the drying process of the non-stoichiometric solvate, a portion of the solvent can be removed without significantly disturbing the crystal network, and the resulting solvate can then be resolvated to obtain the initial crystalline form. Unlike stoichiometric solvates, desolvation and resolvation of non-stoichiometric solvates are not accompanied by phase changes, and all solvation states represent the same crystalline form.

[0078] The term "hydrate" refers to a compound associated with water. Usually, the number of water molecules contained in the hydrate of a compound is in a certain ratio to the number of compound molecules in the hydrate. Thus, a hydrate of a compound can be represented, for example, by the general formula R·xH 2 O represents, where R is a compound and where x is a number greater than 0. A given compound may form more than one type of hydrate, including, for example, a monohydrate (x is 1), a lower hydrate (x is a number greater than 0 and less than 1, such as a hemihydrate (R·0.5H 2 O)) and polyhydrates (x is a number greater than 1, such as dihydrate (R·2H 2 O) and hexahydrate (R·6H 2 O)).

[0079] The term "tautomer" refers to compounds that are interchangeable forms of a particular compound structure and that differ in the replacement of hydrogen atoms and electrons. Thus, the two structures can be in equilibrium by the movement of π electrons and atoms (usually H). For example, enols and ketones are tautomers because they are rapidly interconverted by treatment with an acid or base. Another example of tautomerism is the acid and nitro forms of phenylnitromethane, which are also formed by treatment with an acid or base.

[0080] Tautomeric forms can be relevant to the achievement of optimal chemical reactivity and biological activity of the compound of interest. The exact ratio of tautomers depends on several factors, including temperature, solvent, and pH. Tautomerization (i.e., a reaction providing a tautomeric pair) can be catalyzed by acid or base. Exemplary tautomerizations include keto-to-enol, amide-to-imide, lactam-to-lactim, enamine-to-imine, and enamine-to-(different enamine) tautomerizations.

[0081] It is also understood that compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed "isomers." Isomers that differ in the arrangement of their atoms in space are termed "stereoisomers."

[0082] Stereoisomers that are not mirror images of each other are called "diastereomers", while stereoisomers that are non-superimposable mirror images of each other are called "enantiomers". When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers may occur. Enantiomers can be characterized by the absolute configuration of their asymmetric center, and described by the R- and S-ordering rules of Cahn and Prelog, or by the way the molecule rotates the plane of polarized light, and are designated as right- or left-handed (i.e., (+) or (-)-isomers, respectively). Chiral compounds can exist as individual enantiomers or as mixtures thereof. A mixture containing equal proportions of enantiomers is called a "racemic mixture".

[0083] The term "crystalline" or "crystalline form" refers to a solid form that exhibits substantially three-dimensional order. In certain embodiments, the crystalline form of a solid is a substantially non-amorphous solid form. In certain embodiments, the X-ray powder diffraction (XRPD) pattern of the crystalline form comprises one or more clearly defined peaks.

[0084] The term "amorphous" or "amorphous form" refers to a solid form ("solid form") that is substantially lacking in three-dimensional order. In certain embodiments, the amorphous form of a solid is a substantially non-crystalline solid form. In certain embodiments, using CuKα radiation, the X-ray powder diffraction (XRPD) pattern of the amorphous form includes a broad scattering band with a peak at 2θ of, for example, 20 to 70 ° (including end values) of the bracket. In certain embodiments, the XRPD pattern of the amorphous form further includes one or more peaks attributed to a crystalline structure. In certain implementations, the maximum intensity of any one of the one or more peaks attributed to a crystalline structure observed at 2θ between 20 to 70 ° (including end values) is no more than 300 times, no more than 100 times, no more than 30 times, no more than 10 times, or no more than 3 times the maximum intensity of the broad scattering band. In certain embodiments, the XRPD pattern of the amorphous form does not include a peak attributed to a crystalline structure.

[0085] The term "cocrystal" refers to a crystalline structure comprising at least two different components (e.g., a compound disclosed herein and an acid), wherein each component is independently an atom, an ion, or a molecule. In certain embodiments, none of the components is a solvent. In certain embodiments, at least one component is a solvent. The cocrystal of a compound disclosed herein and an acid is different from a salt formed by a compound disclosed herein and an acid. In a salt, a compound disclosed herein is complexed with an acid in a manner that a proton is easily transferred from an acid to a compound disclosed herein (e.g., complete proton transfer) at room temperature. However, in a cocrystal, a compound disclosed herein is complexed with an acid in a manner that a proton is not easily transferred from an acid to a compound disclosed herein at room temperature. In certain embodiments, in a cocrystal, no proton is transferred from an acid to a compound disclosed herein. In certain embodiments, in a cocrystal, some protons are transferred from an acid to a compound disclosed herein. Cocrystals can be used to improve the properties (e.g., solubility, stability, and ease of formulation) of a compound disclosed herein.

[0086] The term "polymorph" refers to a crystalline form of a compound (or its salt, hydrate or solvate) in a specific crystal packing arrangement. All polymorphs have the same elemental composition. Different crystalline forms typically have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardnesses, crystal shapes, optical and electrical properties, stability and solubility. Recrystallization solvents, crystallization rates, storage temperatures and other factors may lead to a dominant crystalline form. Various polymorphs of a compound can be prepared by crystallization under different conditions.

[0087] The term "prodrug" refers to a compound, including a derivative of compound (I), which has a cleavable group and becomes a compound (I) that is pharmaceutically active in vivo by solvolysis or under physiological conditions. Such examples include, but are not limited to, ester derivatives, etc. Other derivatives of the disclosed compounds are active in both their acid and acid derivative forms, but generally provide advantages of solubility, tissue compatibility or delayed release in mammalian organisms in acid-sensitive forms (see, Bundgard, H., "Design of Prodrugs", pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs include acid derivatives well known to those skilled in the art, such as esters prepared by reacting the parent acid with a suitable alcohol, or amides prepared by reacting the parent acid compound with a substituted or unsubstituted amine, or anhydrides, or mixed anhydrides. Simple aliphatic or aromatic esters, amides and anhydrides derived from the acidic groups attached to the disclosed compounds are specific prodrugs. In some cases, it is desirable to prepare double ester type prodrugs, such as (acyloxy)alkyl esters or ((alkoxycarbonyl)oxy)alkyl esters.

[0088] "Subjects" contemplated for administration include, but are not limited to, humans (i.e., males or females of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or elderly)) and / or other non-human animals, e.g., mammals (e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys); commercially relevant mammals such as cattle, pigs, horses, sheep, goats, cats, and / or dogs) and birds (e.g., commercially relevant birds such as chickens, ducks, geese, and / or turkeys). In certain embodiments, the animal is a mammal. The animal can be male or female and at any stage of development. The non-human animal can be a transgenic animal. A subject resistant to treatment with a BTK inhibitor is a subject that has no response or minimal response to treatment. In some embodiments, the response to treatment is measured by tumor cell reduction or tumor cell killing. In some embodiments, the response to treatment is measured by changes in symptoms of a disease, disorder, or malignancy (e.g., a proliferative disease). It has been discovered that compounds that block ATP binding to HCK as described herein are able to cause tumor cell killing even in cells derived from subjects that are resistant to treatment with BTK inhibitors.

[0089] The terms "administer," "administering," or "administration" refer to implanting, absorbing, ingesting, injecting, inhaling, or otherwise introducing a compound or pharmaceutical composition thereof.

[0090] The terms "treatment", "treat" and "treating" refer to reversing, alleviating, delaying the onset of, or inhibiting the progression of, a "pathological condition" (e.g., a disease, disorder or condition, or one or more signs or symptoms thereof) described herein. In some embodiments, treatment may be administered after one or more signs or symptoms have developed or have been observed. In other embodiments, treatment may be administered in the absence of signs or symptoms of a disease or condition. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., based on a history of symptoms and / or based on genetic or other susceptibility factors). Treatment may also be continued after symptoms have resolved, e.g., to delay or prevent recurrence.

[0091] The terms "condition," "disease," and "disorder" are used interchangeably. Treatment may be therapeutic treatment (and does not include prophylactic or preventative treatment).

[0092] An "effective amount" of Compound (I) is an amount sufficient to elicit the desired biological response, i.e., to treat a condition. As will be appreciated by one of ordinary skill in the art, the effective amount of Compound (I) may vary depending on factors such as the desired biological endpoint, the pharmacokinetics of the compound, the condition being treated, the mode of administration, and the age and health of the subject. An effective amount encompasses both therapeutic and prophylactic treatments. For example, in the treatment of cancer, an effective amount of the compound may reduce tumor burden or prevent the growth or spread of a tumor.

[0093] A "therapeutically effective amount" of compound (I) is an amount sufficient to provide a therapeutic benefit in the treatment of a disorder or to delay or minimize one or more symptoms associated with the disorder. A therapeutically effective amount of a compound means an amount of a therapeutic agent, alone or in combination with other therapies, that provides a therapeutic benefit in the treatment of a disorder. The term "therapeutically effective amount" may encompass an amount that improves overall therapy, reduces or avoids symptoms or causes of a disorder, or enhances the therapeutic effect of another therapeutic agent.

[0094] A "prophylactically effective amount" of compound (I) is an amount sufficient to prevent a disorder or one or more symptoms associated with the disorder or to prevent its recurrence. A prophylactically effective amount of a compound means an amount of a therapeutic agent, alone or in combination with other agents, that provides a prophylactic benefit in the prevention of a disorder. The term "prophylactically effective amount" may encompass an amount that improves overall prevention or enhances the prophylactic effect of another prophylactic agent.

[0095] "Proliferative disease" refers to a disease that occurs due to abnormal growth or expansion of cellular proliferation (Walker, Cambridge Dictionary of Biology; Cambridge University Press: Cambridge, England, 1990). Proliferative diseases may be associated with: 1) pathological proliferation of normally dormant cells; 2) pathological migration of cells from their normal location (e.g., metastasis of tumor cells); 3) pathological expression of proteolytic enzymes, such as matrix metalloproteinases (e.g., collagenases, gelatinases, and elastases); or 4) pathological angiogenesis, as in proliferative retinopathy and tumor metastasis. Exemplary proliferative diseases include cancer (e.g., breast cancer, colon cancer, testicular cancer, CNS cancer, gastric cancer, lymphoma (e.g., B cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., secreting IgM (i.e., Waldenstrom's macroglobulinemia)), non-secreting IgM)), diffuse large B cell lymphoma (e.g., activated B cell-like (ABC)-DLBCL, germinal center B cell-like (GBC)-DLBCL)), follicular lymphoma, marginal zone B cell lymphoma, small lymphocytic lymphoma (e.g., chronic lymphocytic leukemia (CLL)), mantle cell lymphoma), leukemia (e.g., myeloid leukemia (e.g., chronic myeloid leukemia, acute myeloid leukemia)), benign neoplasms, angiogenesis, inflammatory diseases, autoinflammatory diseases, and autoimmune diseases.

[0096] The terms "vegetation" and "tumor" can be used interchangeably and refer to an abnormal tissue mass, wherein the growth of the mass exceeds the growth of normal tissue and is not coordinated with the growth of normal tissue. Vegetation or tumor can be "benign" or "malignant", depending on the following characteristics: degree of cell differentiation (including morphology and function), growth rate, local invasion and metastasis. "Benign neoplasms" are usually well differentiated, and their characteristic growth is slower than that of malignant tumors, and are confined to the primary site. In addition, benign neoplasms do not have the ability to infiltrate, invade or metastasize to distant sites. Exemplary benign neoplasms include but are not limited to lipomas, chondromas, adenomas, soft droopy warts, senile hemangiomas, seborrheic keratosis, freckles and sebaceous hyperplasia. In some cases, some "benign" tumors may subsequently develop into malignant neoplasms, which may be caused by additional genetic changes in a subpopulation of the neoplastic cells of the tumor, and these tumors are referred to as "premalignant neoplasms". Exemplary premalignant neoplasms are teratomas. In contrast, "malignant neoplasms" are usually poorly differentiated (anaplastic) and are characterized by rapid growth, accompanied by progressive infiltration, invasion, and destruction of surrounding tissue. In addition, malignant neoplasms often have the ability to metastasize to distant sites.

[0097] The terms "metastasis," "metastatic," or "metastasize" refer to the spread or migration of cancer cells from a primary or original tumor to another organ or tissue, and can often be identified by the presence of a "secondary tumor" or "secondary cell population" of the tissue type of the primary or original tumor rather than the organ or tissue in which the secondary (metastatic) tumor is located. For example, prostate cancer that has migrated to the bone is called metastatic prostate cancer and contains cancerous prostate cancer cells growing in the bone tissue.

[0098] The term "cancer" refers to a malignant neoplasm (Stedman's Medical Dictionary, 25th ed.; Hensyl, ed.; Williams & Wilkins: Philadelphia, 1990). Cancer can be a solid tumor. Cancer can be a hematological malignancy. Exemplary cancers include, but are not limited to, acoustic neuroma; adenocarcinoma; adrenal cancer; anal cancer; angiosarcoma (e.g., lymphangiosarcoma, lymphangioendothelioma, hemangioendothelioma); appendix cancer; benign monoclonal gammopathy; bile duct cancer (e.g., hepatobiliary carcinoma); bladder cancer; breast cancer (e.g., breast adenocarcinoma, breast papillary carcinoma, breast carcinoma, medullary breast carcinoma); brain cancer (e.g., meningioma, glioblastoma, glioma (e.g., astrocytoma, oligodendroglioma), medulloblastoma); bronchogenic carcinoma; carcinoid tumor; cervical cancer (e.g., cervical adenocarcinoma); choriocarcinoma; chordoma; craniopharyngioma; colorectal cancer (e.g., colon cancer, rectal cancer, colorectal adenocarcinoma); connective tissue cancer; epithelial cancer; ependymoma; endothelial sarcoma (e.g., Kaposi sarcoma, multiple idiopathic hemorrhagic sarcoma); endometrial cancer (e.g., uterine cancer, uterine sarcoma); Esophageal cancer (e.g., esophageal adenocarcinoma, Barrett's adenocarcinoma); Ewing's sarcoma; eye cancer (e.g., intraocular melanoma, retinoblastoma); familial eosinophilia; gallbladder cancer; gastric cancer (e.g., gastric adenocarcinoma); gastrointestinal stromal tumor (GIST); germ cell cancer; head and neck cancer (e.g., head and neck squamous cell carcinoma, oral cancer (e.g., oral squamous cell carcinoma), pharyngeal cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngeal carcinoma, oropharyngeal cancer)); hematopoietic cancer (e.g., leukemia, such as acute lymphoblastic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acute myeloid leukemia (AML) (e.g., B-cell AML, T-cell AML), chronic myeloid leukemia (CML) (e.g., B-cell CML, T-cell CML) and chronic lymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL));Lymphoma, such as Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) and non-Hodgkin lymphoma (NHL) (e.g., B-cell NHL, such as diffuse large cell lymphoma (DLCL) (e.g., diffuse large B-cell lymphoma (DLBCL) (e.g., activated B-cell-like (ABC)-DLBCL, germinal center B-cell-like (GBC)-DLBCL)), follicular lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma (CLL / SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphoma (e.g., mucosa-associated lymphoid tissue (MALT) lymphoma, nodal marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma lymphoma, Burkitt's lymphoma, lymphoplasmacytic lymphoma (e.g., IgM-secreting lymphoplasmacytic lymphoma, i.e., Waldenstrom's macroglobulinemia, non-IgM-secreting lymphoplasmacytic lymphoma), hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B lymphoblastic lymphoma, and primary central nervous system (CNS) lymphoma; and T-cell NHL, such as precursor T-lymphoblastic lymphoma / leukemia, peripheral T-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g., mycosis fungoides, Sezary syndrome), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy-type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, and anaplastic large cell lymphoma); a mixture of one or more of the leukemias / lymphomas described above; and multiple myeloma (MM)), heavy chain disease (e.g., alpha chain disease, gamma chain disease, mu chain disease); hemangioblastoma; hypopharyngeal cancer; inflammatory myofibroblastic tumor; immunocytic amyloidosis; kidney cancer (e.g., Wilms' tumor, renal cell carcinoma); liver cancer (e.g., hepatocellular carcinoma (HCC), malignant hepatoma); lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), lung adenocarcinoma); leiomyosarcoma (LMS); mastocytosis (e.g., systemic mastocytosis); muscle cancer; myelodysplastic syndrome (MDS); mesothelioma myeloproliferative disorders (MPDs) (e.g., polycythemia vera (PV), essential thrombocythemia (ET), myeloid metaplasia of unknown etiology (AMM) (also known as myelofibrosis (MF)), chronic idiopathic myelofibrosis, chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES)); neuroblastoma; neurofibromas (e.g., neurofibromatosis type 1 or type 2 (NF), schwannomatosis); neuroendocrine cancers (e.g., gastroenteropancreatic neuroendocrine tumors (GEP-NET), carcinoid tumors); osteosarcomas (e.g., bone cancer); ovarian cancers (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma); papillary adenocarcinoma;Pancreatic cancer (e.g., pancreatic adenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), islet cell tumor); penile cancer (e.g., Paget's disease of the penis and scrotum); pinealoma; primitive neuroectodermal tumor (PNT); plasmacytoma; paraneoplastic syndrome; intraepithelial neoplasia; prostate cancer (e.g., prostate adenocarcinoma); rectal cancer; rhabdomyosarcoma; salivary gland cancer; skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)); small intestine cancer of the testis (e.g., seminoma, embryonal carcinoma of the testis); cancer of the thyroid gland (e.g., papillary thyroid carcinoma, papillary thyroid carcinoma (PTC), medullary thyroid carcinoma); cancer of the urethra; cancer of the vagina; and cancer of the vulva (e.g., Paget's disease of the vulva).

[0099] The term "angiogenesis" refers to the formation and growth of new blood vessels. Normal angiogenesis occurs in the healthy body of a subject for healing wounds and for restoring blood flow to tissues after injury. A healthy body controls angiogenesis in a variety of ways, such as angiogenesis stimulating growth factors and angiogenesis inhibitors. Many disease states, such as cancer, diabetic blindness, age-related macular degeneration, rheumatoid arthritis and psoriasis, are characterized by abnormal (i.e., increased or excessive) angiogenesis. Abnormal or pathological angiogenesis refers to angiogenesis greater than that in a normal body, especially angiogenesis in adults that is unrelated to normal angiogenesis (e.g., menstruation or wound healing). Abnormal angiogenesis can provide new blood vessels to nourish diseased tissues and / or destroy normal tissues, and in the case of cancer, new blood vessels can allow tumor cells to escape into the circulatory system and stay in other organs (tumor metastasis). In certain embodiments, angiogenesis is pathological angiogenesis.

[0100] "Autoimmune disease" refers to a disease caused by an inappropriate immune response of the subject's body to substances and tissues normally present in the body. In other words, the immune system mistakes some part of the body for a pathogen and attacks its own cells. This may be localized to certain organs (e.g., in autoimmune thyroiditis) or involve specific tissues in different locations (e.g., Goodpasture's disease, which may affect the basement membranes of the lungs and kidneys). Treatment of autoimmune diseases often uses immunosuppression, such as medications that reduce the immune response. Exemplary autoimmune diseases include, but are not limited to, glomerulonephritis, Goodpasture's syndrome, necrotizing vasculitis, lymphadenitis, periarteritis nodosa, systemic lupus erythematosus, rheumatoid arthritis, psoriatic arthritis, systemic sclerosis, psoriasis, ulcerative colitis, systemic sclerosis, dermatomyositis / polymyositis, antiphospholipid antibody syndrome, scleroderma, pemphigus vulgaris, ANCA-associated vasculitis (e.g., Wegener's granulomatosis, microscopic polyangiitis), uveitis, Sjogren's syndrome, Crohn's disease, Reiter's syndrome, ankylosing spondylitis, Lyme disease, Guillain-Barré syndrome, Hashimoto's thyroiditis, and cardiomyopathy.

[0101] The term "inflammatory disease" refers to a disease caused by, caused by, or resulting in inflammation. The term "inflammatory disease" may also refer to an inflammatory response that is dysregulated, causing an over-response of macrophages, granulocytes, and / or T lymphocytes, leading to abnormal tissue damage and / or cell death. Inflammatory diseases may be acute or chronic inflammatory conditions and may be caused by infection or non-infectious causes. Inflammatory diseases include atherosclerosis, arteriosclerosis, autoimmune diseases, multiple sclerosis, systemic lupus erythematosus, polymyalgia rheumatica (PMR), gouty arthritis, degenerative arthritis, tendinitis, bursitis, psoriasis, cystic fibrosis, osteoarthritis, rheumatoid arthritis, inflammatory arthritis, Sjögren's syndrome, giant cell arteritis, progressive systemic sclerosis (scleroderma), ankylosing spondylitis, polymyositis, dermatomyositis, pemphigus, pemphigoid, diabetes (e.g., type I), myasthenia gravis, Hashimoto's thyroiditis, Graves' disease, Goodpasture's disease, mixed Connective tissue disease, sclerosing cholangitis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, pernicious anemia, usual interstitial pneumonia (UIP), asbestosis, silicosis, bronchiectasis, berylliosis, talcosis, pneumoconiosis, sarcoidosis, desquamative interstitial pneumonia, lymphointerstitial pneumonia, giant cell interstitial pneumonia, cellular interstitial pneumonia, extrinsic allergic alveolitis, Wegener's granulomatosis and related forms of vasculitis (temporal arteritis and polyarteritis nodosa), inflammatory skin diseases, dermatitis (e.g., stasis dermatitis, allergic contact dermatitis, atopic dermatitis, irritant contact dermatitis, neurodermatitis, perioral dermatitis, seborrheic dermatitis, dermatitis), hepatitis, delayed hypersensitivity reactions (e.g., poison ivy dermatitis), pneumonia, respiratory tract inflammation, adult respiratory distress syndrome (ARDS), encephalitis, immediate hypersensitivity reactions, asthma, hay fever, allergy, acute anaphylaxis, rheumatic fever, glomerulonephritis, pyelonephritis, cellulitis, cystitis, chronic cholecystitis, ischemia (ischemic injury), reperfusion injury, allograft rejection, host-versus-graft rejection, appendicitis, arteritis, blepharitis, bronchiolitis, bronchitis, cervicitis, cholangitis, chorioamnionitis, conjunctivitis, dacryoadenitis, dermatomyositis, endocarditis, endometritis, enteritis, epicondylitis, Epididymitis, fasciitis, fibrositis, gastritis, gastroenteritis, gingivitis, ileitis, iritis, laryngitis, myelitis, myocarditis, nephritis, omphalitis, oophoritis, orchitis, osteitis, otitis, pancreatitis, mumps, pericarditis, pharyngitis, pleurisy, phlebitis, pneumonia, proctitis, prostatitis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis, orchitis, tonsillitis, urethritis, cystitis, uveitis, vaginitis, vasculitis, vulvitis, vulvovaginitis, vasculitis, chronic bronchitis, osteomyelitis, optic neuritis, temporal arteritis, transverse myelitis, necrotizing fasciitis, necrotizing enterocolitis, inflammatory rosacea. Inflammatory diseases of the eye include postoperative inflammation.

[0102] The term "kinase" refers to any enzyme that catalyzes the addition of a phosphate group to an amino acid residue of a protein. For example, a serine kinase catalyzes the addition of a phosphate group to a serine residue of a protein. In certain embodiments, the kinase is a protein kinase. Examples of kinases include, but are not limited to, cytoplasmic tyrosine kinases (e.g., SRC family kinases (e.g., HCK, LYN, BLK, FRK), Tec family kinases (e.g., BTK)), cyclin-dependent kinases (CDKs, e.g., CDK1, CDK2, CDK2, CDK4, CDK5, CDK7, CDK8, CDK9, CDK10, CDK11, CDK12, CDK13, CDK14, CDK16, CDK20)), mitogen-activated protein kinases (MAPKs, e.g., MAP K1, MAPK3, MAPK4, MAPK6, MAPK7, MAPK8, MAPK9, MAPK10, MAPK11, MAPK12, MAPK13, MAPK14, MAPK15), glycogen synthase kinase 3 (GSK3, e.g., GSK3α, GSK3β), CDK-like kinases (CLK, e.g., CLK1, CLK2, CLK3, CLK4)), AGC kinases (e.g., protein kinase A (PKA), protein kinase C (PKC), protein kinase G (PKG)), Ca 2+ / calmodulin-dependent protein kinase (CaM kinase, e.g., specialized CaM kinase, multifunctional CaM kinase), casein kinase 1 (CK1, e.g., CK1α, CK1β1, Ck1γ1, Ck1γ2, Ck1γ3, CK1δ, CK1ε), STE kinase (e.g., homologs of yeast sterile 7, sterile 11 or sterile 20 kinase), tyrosine kinase (TK, e.g., receptor tyrosine kinase (RTK), non-receptor tyrosine kinase (nRTK)) and tyrosine kinase-like kinase (TKL, e.g., mixed lineage kinase (MLK), RAF, serine threonine kinase receptor (STKR), leucine-rich repeat kinase (LRRK), LIM domain kinase (LIMK), testis expressed serine kinase (TESK), IL1 receptor-associated kinase (IRAK), receptor interacting protein kinase (RIPK)).

[0103] Hematopoietic cell kinase (HCK) is a member of the src family of protein tyrosine kinases and is abnormally upregulated in WM cells. In myeloma cells, HCK is activated by interleukin 6 (IL6) through the IL6 co-receptor IL6ST (GP130).

[0104] Bruton's tyrosine kinase (BTK), a member of the src-related BTK / Tec family of cytoplasmic tyrosine kinases, is required for B-cell receptor signaling, plays a key role in B-cell maturation, and exhibits increased activation in many B-cell malignancies.

[0105] LYN proto-oncogene (LYN) is a member of the src family of protein tyrosine kinases, plays an important role in regulating B cell differentiation, proliferation, survival and apoptosis, plays an important role in immune self-tolerance, and acts downstream of several immune receptors, including the B cell receptor (BCR). Without wishing to be bound by theory, BCR signaling is believed to be involved in growth-promoting and survival signaling in MYD88 mutant diseases, as well as in non-MYD88 mutant diseases. For example, BCR signaling is believed to be active in Waldenstrom's macroglobulinemia, the ABC subtype of diffuse large B cell lymphoma, and chronic lymphocytic leukemia.

[0106] The proto-oncogene tyrosine protein kinase SRC (SRC) is a protein tyrosine kinase that plays an important role in the regulation of multiple biological processes such as proliferation, migration, adhesion and survival of solid tumor cells, and is overexpressed in WM.

[0107] As used herein, "inhibition," "inhibiting," "inhibit," and "inhibitor" and the like refer to the ability of a compound to reduce, slow, stop, block, or prevent the activity of a particular biological process (e.g., a kinase (e.g., SFK (e.g., HCK, LYN, BLK, FRK), Tec family kinase (e.g., BTK)) in a cell relative to the vector.

[0108] The term "block" or "blocking" refers to the ability of a compound to prevent biological interactions (e.g., binding) in a cell relative to a negative control (e.g., vehicle). For example, a compound can block the binding of ATP to the ATP binding pocket of a kinase. This blocking can occur by direct binding of the compound to the ATP binding pocket itself or by indirect blocking. In some embodiments, the term refers to a reduction in the level of binding of ATP to a kinase (e.g., BTK and / or HCK and / or LYN and / or SRC) to a level that is statistically significantly lower than the initial level, which initial level can be, for example, a baseline level of ATP binding. In some embodiments, the term refers to a reduction in the level of ATP binding to a kinase (e.g., BTK and / or HCK, and / or LYN, and / or SRC) to a level that is less than 75%, less than 50%, less than 40%, less than 30%, less than 25%, less than 20%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.01%, less than 0.001%, or less than 0.0001% of the initial level, which initial level can be, for example, a baseline level of ATP binding. In some embodiments, blocking ATP binding results in a reduction in the level of enzyme activity (e.g., BTK and / or HCK and / or LYN and / or SRC activity) to a level that is less than 75%, less than 50%, less than 40%, less than 30%, less than 25%, less than 20%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.01%, less than 0.001%, or less than 0.0001% of the initial level, which initial level can be, for example, a baseline level of enzyme activity.

[0109] When a compound or pharmaceutical composition is said to "selectively," "specifically," or "competitively" bind to a first protein, the compound binds to the first protein (e.g., BTK or HCK or LYN or SRC) with a higher binding affinity (e.g., not less than about 2-fold, not less than about 5-fold, not less than about 10-fold, not less than about 30-fold, not less than about 100-fold, not less than about 1,000-fold, or not less than about 10,000-fold) than it binds to a second protein (e.g., BTK) that is different from the first protein. In some embodiments, the compound blocks ATP binding to the first protein (e.g., HCK or LYN or SRC) at a concentration that is lower than that at which it blocks ATP binding to the second protein (e.g., BTK) that is different from the first protein (e.g., BTK) (e.g., not less than about 2-fold, not less than about 5-fold, not less than about 10-fold, not less than about 30-fold, not less than about 100-fold, not less than about 1,000-fold, or not less than about 10,000-fold).

[0110] Compounds provided herein that selectively block ATP binding to kinases (e.g., BTK, HCK, LYN) can be identified and / or characterized by methods known in the art. Methods include purified enzyme and cell-based biochemical and binding assays, such as HCK gatekeeper mutant rescue assays, in vitro kinase assays, such as using HCK gatekeeper mutant kinases, using KiNativ TM Suitable assays for determining the selective inhibition of HCK by a compound include, but are not limited to, the Life Technology Z-Lyte activity assay (e.g., including HCK gatekeeper mutants and GK + 6 mutants); DiscoverX Binding assay; MRC radioactivity assay; ACD Ba / F3 viability assay (e.g., containing HCK gatekeeper mutants and GK + 6 mutants); yeast hybrid proliferation assay; protein thermal stability assay; and HCK gatekeeper mutants or GK + Proliferation-rescue assay of cancer cells with mutants of LYN-6. Such assays can also be used to determine the selective inhibition of LYN and / or SRC by compounds.

[0111] The term "MYD88 mutation" means any change or difference in the nucleic acid or protein sequence of MYD88 compared to the wild-type sequence, which leads to the activation of MYD88, thereby leading to the activation of NF-κB. Mutations include but are not limited to nonsense mutations, missense mutations, frameshift mutations, rearrangement mutations, insertion mutations and deletion mutations. In some embodiments, the mutation is a somatic mutation at position 38182641 in chromosome 3p22.2, which results in a single nucleotide change of T→C in the primary response to myeloid differentiation (MYD88) gene, and a non-synonymous change from leucine to proline (L265P) predicted at amino acid position 265. In some embodiments, the mutation is another activating mutation in MYD88, such as V217F, W218R, I220T, S222R, M232T, S243N, T294P. Signaling studies have shown that SU-DHL-2 lymphoma cells expressing a serine to arginine mutation at amino acid position 222 also have upregulated HCK (Yang et al., Blood 2016). In some embodiments, Sanger sequencing, whole exome or whole genome sequencing can be used to identify somatic mutations in MYD88.

[0112] The term "MYD88 mutation disease" or "disease associated with mutated MYD88" means any disease associated with changes or differences in the nucleic acid or protein sequence of MYD88 compared to the wild-type sequence in a subject, which results in activation of MYD88, thereby resulting in activation of NF-κB. In some embodiments, mutated MYD88 is associated with Waldenstrom's macroglobulinemia (IgM-secreting lymphoplasmacytic lymphoma), non-IgM-secreting lymphoplasmacytic lymphoma, ABC subtype of diffuse large B-cell lymphoma, primary central nervous system (CNS) lymphoma, immune privileged lymphoma including testicular lymphoma, marginal zone lymphoma, and chronic lymphocytic leukemia. In some embodiments, mutated MYD88 is associated with susceptibility to infectious diseases. In some embodiments, mutated MYD88 is associated with susceptibility to autoimmune diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0113] Figure 1 The kinases that may be targeted by compound (I) are shown.

[0114] FIG2 shows that compound (I) selectively targets HCK, BTK and LYN in MYD88-mutated WM and ABC DLBCL cells. Figure 2A Compound (I) and A419259 are shown against a panel of 464 kinases. result. Figure 2B The KiNativ of compound (I) is shown TM Cell Target Binding Curves. Kinases that were treated by more than 50% with 1.0 μM Compound (I) in TMD8 cells are listed. Figure 2C Shown are activated kinases enriched by pull-down assay using desthiobiotin-ATP probe after pretreatment of BCWM.1 and TMD8 live cells with indicated concentrations of compound (I) or ibrutinib for 90 min, and ATP-bound HCK, BTK and LYN resolved by Western blotting.

[0115] FIG3 shows the effects of compound (I) and ibrutinib on MYD88 mutant WM (BCWM.1, MWCL-1) and ABC DLBCL (TMD8, HBL-1) cell lines by PhosFlow analysis (N=3) ( Figure 3A ) and by Western blotting ( Figure 3B ) ; and HCK and BTK phosphorylation analyzed by PhosFlow in bone marrow tumor cells of WM patients with MYD88 mutations (N=4) ( Figure 3C )’s impact.

[0116] FIG4 shows the effect of compound (I) on MYD88 wild type (MYD88 WT) and MYD88 mutation (MYD88 L265P ) Cellular efficacy in B-cell lymphoma cells. MYD88 WT and MYD88 L265P Dose response of cell lines after treatment with serially diluted compound (I), A419259 and ibrutinib for 72 hours. Figure 4A ). Apoptotic activity of primary BM lymphoplasmacytic cells from CD19+ B cells of 6 MYD88 mutation patients and 6 healthy donors after 16 h of treatment with the indicated concentrations of ibrutinib or compound (I). *p<0.05; **p<0.01; ****p<0.0001.

[0117] Figure 5 shows that HCK gatekeeper mutant (HCK T333M ) rescued compound (I)-induced cell death and blocked HCK activation and its downstream signaling in BCWM.1WM cells. Figure 5A Shown are vehicle or HCK only WT or HCK T333M Dose response of transduced BCWM.1 cells after 72 hours of compound (I) treatment. Figure 5B Shown in the case of vector only, HCK WT or HCK T333M Relative pHCK resolved by PhosFlow analysis in transduced MYD88 mutant BCWM.1 cells treated with indicated concentrations of compound (I) for 1.0 h Y411 level. Figure 5C The results show that HCK WT or HCK T333M pBTK in transduced BCWM.1 cells analyzed by western blot Y223 , pSYK Y525 / 526 , p-AKT S473 、p-ERK1 / 2 T202 / Y204 Also shown are the expression levels of total BTK, SYK, AKT and ERK1 / 2 in these cells, as well as the protein loading control GAPDH.

[0118] Figure 6 shows that HCK gatekeeper mutant (HCK T333M) Rescue compound (I)-induced cell death and block HCK activation and its downstream signaling in MWCL-1WM cells. Fig. 6A Shown are vehicle or HCK only WT or HCK T333M Dose response of transduced BCWM.1 cells after 72 hours of compound (I) treatment. Figure 6B Shown in the case of vector only, HCK WTor HCK T333M Relative pHCK resolved by PhosFlow analysis in transduced MYD88 mutant BCWM.1 cells treated with indicated concentrations of compound (I) for 1.0 h Y411 level. Figure 6C The results show that HCK WT or HCK T333M pBTK in transduced BCWM.1 cells analyzed by western blot Y223 , pSYK Y525 / 526 , p-AKT S473 、p-ERK1 / 2 T202 / Y204 Also shown are the expression levels of total BTK, SYK, AKT and ERK1 / 2 in these cells, as well as the protein loading control GAPDH.

[0119] FIG. 7 shows that the HCK gatekeeper mutant (HCK T333M ) rescued compound (I)-induced cell death and blocked HCK activation and its downstream signaling in TMD8ABC DLBCL cells. Fig. 7A Shown are vehicle or HCK only WT or HCK T333M Dose response of transduced BCWM.1 cells after 72 hours of compound (I) treatment. Figure 7B Shown in the case of vector only, HCK WT or HCK T333M Relative pHCK resolved by PhosFlow analysis in transduced MYD88 mutant BCWM.1 cells treated with indicated concentrations of compound (I) for 1.0 h Y411 level. Figure 7C The results show that HCK WT or HCK T333M pBTK in transduced BCWM.1 cells analyzed by western blot Y223 , pSYK Y525 / 526 , p-AKT S473 、p-ERK1 / 2 T202 / Y204 Also shown are the expression levels of total BTK, SYK, AKT and ERK1 / 2 in these cells, as well as the protein loading control GAPDH.

[0120] Figure 8 The in vitro and in vivo pharmacokinetic properties of Compound (I) are shown.

[0121] Fig. 9 The in vitro and in vivo pharmacokinetic properties of A419259 are shown.

[0122] FIG. 10 shows a pharmacodynamic study showing ibrutinib sensitivity in subcutaneous xenografts of BTK WT Activity of HCK and BTK after oral administration of compound (I) in NOD-SCID mice with TMD8ABC-DLBCL cells. GFP excised 6 and 24 hours after oral administration of compound (I) at the indicated doses (n=3 per group) + pHCK in TMD8 tumor cells Y411 ( Fig. 10A ) and pBTK Y223 ( Fig. 10B ). ****p<0.0001.

[0123] Figure 11 shows the effect of compound (I) on BTK WT Effects on tumor volume and survival in a TMD8 ABC DLBCL xenograft mouse model. Following daily oral administration of 50 mg / kg of vehicle control, ibrutinib, or compound (I) in mice bearing ibrutinib-sensitive BTK WT Efficacy study in NOD-SCID mice (n=8 per group) using TMD8 cells. Tumor volume (mm3) was measured twice a week and reported as mean volume ± SEM. Treatment was stopped on day 42 (indicated by green arrows), and tumor volume was monitored until day 113 ( Fig.11A ). Comparison of tumor volume on day 33. The p-values ​​for group comparisons are shown ( Fig. 11B ). The survival curve was estimated using the Kaplan-Meier method. The median survival of the groups was displayed using Prism software (days). The log-rank comparison between the groups was P < .0001 ( Fig. 11C ).

[0124] FIG12 shows the in vitro cellular efficacy (drug dose-response) of ibrutinib or compound (I). WT or BTK C481S TMD8 ABC-DLBCL or BCWM.1WM cells transduced with lentiviral vectors were expressed by CellTiter-Glo TM Cell viability assays to measure drug dose-response ( Fig. 12A We measured apoptosis analysis by flow cytometry using Annexin-V / PI staining, marking the percentage of apoptotic cells ( Fig. 12B ).

[0125] Figure 13 shows that compound (I) blocks HCK, BTK and downstream ERK1 / 2 activation in BCWM.1WM and TMD8 ABC DLBCL cell lines with BTK Cys481 mutation. WT or BTKC481S Relative pHCK resolved by PhosFlow analysis in transduced BCWM.1 and TMD8 cells after treatment with indicated concentrations of ibrutinib or compound (I) for 1.0 h Y411 level( Fig.13A ). In vector-only, BTK WT or BTK C481S pBTK expression in transduced BCWM.1 and TMD8 cells after treatment with indicated concentrations of ibrutinib or compound (I) for 1.0 h by Western blot analysis Y223 and p-ERK1 / 2 T202 / Y204 Expression. The expression levels of total BTK and ERK1 / 2 and the protein loading control GAPDH in these cells are also shown ( Fig. 13B ).

[0126] FIG. 14 shows a pharmacodynamic study showing that BTK C481S Activity of HCK and BTK after oral administration of compound (I) in NOD-SCID mice with TMD8ABC-DLBCL cells. GFP excised 6 and 24 hours after oral administration of compound (I) at the indicated doses (n=3 per group) + pHCK in TMD8 tumor cells Y411 ( Fig.14A ) and pBTK Y223 ( Fig. 14B ). *p<0.05; **p<0.01; ***p<0.005; ****p<0.0001.

[0127] FIG. 15 shows the effect of compound (I) on ibrutinib-resistant BTK C481S Effects on tumor volume and survival in a TMD8 ABC DLBCL xenograft mouse model. Following daily oral administration of vehicle control, ibrutinib (50 mg / kg), or compound (I) (50 or 75 mg / kg), mice carrying a mouse model expressing ibrutinib-resistant BTK C481S Efficacy study in NOD-SCID mice (n=8 per group) with TMD8 cells. Tumor volume (mm 3 ) and reported as mean volume ± SEM ( Fig.15A ). Comparison of tumor volume on day 29. The p-values ​​for group comparisons are shown ( Fig. 15B ). The survival curve was estimated using the Kaplan-Meier method. The median survival of the groups was displayed using Prism software (days). The log-rank comparison between the groups was P < .0007 ( Fig. 15C ).

[0128] FIG. 16 shows the effect of compound (I) on ibrutinib-resistant BTK C481S Effects on tumor volume and survival in a TMD8 ABC DLBCL xenograft mouse model. Following daily oral administration of vehicle control, ibrutinib (50 mg / kg), A419259 (50 mg / kg), or compound (I) (50 mg / kg) in mice carrying a mouse model expressing ibrutinib-resistant BTK C481S Efficacy study in NOD-SCID mice (n=8 per group) with TMD8 cells. Tumor volume (mm 3 ) and reported as mean volume ± SEM ( Fig.16A ). Comparison of tumor volume on day 33. The p-values ​​for group comparisons are shown ( Fig. 16B The survival curve was estimated using the Kaplan-Meier method. The median survival time (days) of the groups was displayed using Prism software. The log-rank comparison between the groups was P<0.0001 ( Fig. 16C ).

[0129] Fig.17 Shown are native (untransduced), BTK WT and BTK C481S In vitro studies evaluating the synergistic interaction of compound (I) and venetoclax in BCWM.1WM and TMD8ABC DLBCL cells expressing MYD88 mutations. Combination index (CI) and normalized isobologram analysis are depicted. CI < 1 (indicated by red shading) or points below the diagonal line in the isobologram indicate a synergistic effect of the combination.

[0130] FIG. 18 shows the effect of the combination of Compound (I) and venetoclax on ibrutinib-resistant BTK C481S Effects on tumor volume and survival in a TMD8 ABCDLBCL xenograft mouse model. Following daily oral administration of vehicle control, venetoclax (50 mg / kg), compound (I) (30 mg / kg), or a combination of venetoclax (50 mg / kg) and compound (I) (30 mg / kg), the tumor volume and survival of mice carrying a TMD8 ABCDLBCL xenograft expressing ibrutinib-resistant BTK C481S Efficacy study in NOD-SCID mice (n=8 per group) with TMD8 cells. Tumor volume (mm 3 ) and reported as mean volume ± SEM ( Fig.18A ). Comparison of tumor volume on day 22. The p-values ​​for group comparisons are shown ( Fig.18B The survival curve was estimated using the Kaplan-Meier method. The median survival time (days) of the groups was displayed using Prism software. The log-rank comparison between the groups was P = 0.0020 ( Fig.18C ).

[0131] Fig.19Shown is the assessment of HCK activation status in CLL patients in primary tumor cells from peripheral blood mononuclear cells (PBMCs) of previously treated or untreated patients.

[0132] Fig. 20 Figure 3 shows the expression of BTK in peripheral blood mononuclear cells (PBMCs) from ibrutinib-relapsed CLL patients treated with ibrutinib or compound (I). C481S Annexin V-PI (propidium iodide) assay of mutant freshly isolated tumor cells.

[0133] Fig.21 The results show that the expression of BTK wild type and BTK C481S Western blot analysis of the phosphorylation levels of BTK, LYN, and ERK1 / ERK2 in mutant BCWM.1 cells after incubation with ibrutinib, dasatinib, A419259, and compound (I) for 1 hour. DETAILED DESCRIPTION

[0134] The present disclosure is based in part on the surprising discovery that subjects treated with compound (I) show increased survival rates compared to subjects treated with other BTK inhibitors (e.g., ibrutinib, A419259). Without wishing to be bound by any theory, compared to other BTK inhibitors (e.g., A419259), compound (I) has a shorter half-life in vivo, which may result in increased survival rates. Compared to other BTK inhibitors (e.g., A419259), compound (I) has a shorter half-life, which may result in reduced toxicity of the compound. In certain embodiments, the half-life in vivo of compound (I) is less than 20% of the half-life of other BTK inhibitors (e.g., A419259). In certain embodiments, the half-life in vivo of compound (I) is less than 10% of the half-life of other BTK inhibitors (e.g., A419259).

[0135] Provided herein are methods for treating a disease (e.g., a proliferative disease (e.g., an IgM gammopathy (e.g., IgM monoclonal gammopathy of undetermined significance (MGUS), amyloid light chain (AL) amyloidosis), a mastocytosis (e.g., systemic mastocytosis), a cancer (e.g., breast cancer, colon cancer, testicular cancer, CNS cancer, gastric cancer, a lymphoma (e.g., a B-cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgM-secreting lymphoplasmacytic lymphoma (i.e., Waldenstrom's macroglobulinemia), a non-IgM-secreting lymphoplasmacytic lymphoma)), a diffuse large B-cell lymphoma (e.g., activated B A method for treating a subject with a compound of formula (I) comprising administering to the subject a therapeutically effective amount of:

[0136]

[0137] Or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, isotope-labeled derivative, stereoisomer or prodrug thereof. In certain embodiments, the method comprises administering compound (I) or a pharmaceutically acceptable salt thereof. In certain embodiments, the disease is associated with a mutated MYD88 protein. In certain embodiments, the disease is associated with a mutated BTK protein. In certain embodiments, the disease is associated with a C481 mutated BTK protein. In certain embodiments, the disease is associated with a C481S mutated BTK protein. In some embodiments, the disease is associated with abnormal activity of a kinase (e.g., a SRC family kinase (e.g., HCK, LYN, BLK, FRK), a Tec family kinase (e.g., BTK). In certain embodiments, the disease is resistant to inhibition by a BTK inhibitor (e.g., ibrutinib, CC-292, ONO-4059, evobrutinib, spebrutinib, BGB-3111, HM71224, or ACP-196). In certain embodiments, the disease is resistant to treatment with ibrutinib. In certain embodiments, the disease is associated with a mutated BTK protein (e.g., C481S mutated BTK), and the disease is resistant to treatment with ibrutinib.

[0138] Further provided herein is a method of inhibiting kinase activity in a subject, comprising administering to the subject a therapeutically effective amount of a compound of formula (I):

[0139]

[0140] Or its pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, isotope-labeled derivative, stereoisomer or prodrug. In certain embodiments, the kinase is a SRC family kinase (e.g., HCK, LYN, BLK, FRK). In other embodiments, the kinase is a Tec family kinase (e.g., BTK). In certain embodiments, BTK is resistant to inhibition by BTK inhibitors (e.g., ibrutinib, CC-292, ONO-4059, evobutinib, septinib, BGB-3111, HM71224 or ACP-196). In certain embodiments, BTK is ibrutinib-resistant.

[0141] Provided herein are methods of treating a subject, comprising administering to a subject having a MYD88 mutation disease. The MYD88 mutation disease may include, but is not limited to, a proliferative disease (e.g., IgM gammopathy (e.g., IgM monoclonal gammopathy of undetermined significance (MGUS), amyloid light chain (AL) amyloidosis), mastocytosis (e.g., systemic mastocytosis), cancer (e.g., breast cancer, colon cancer, testicular cancer, CNS cancer, gastric cancer, lymphoma (e.g., B cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgM secreting lymphoplasmacytic lymphoma (i.e., Waldenstrom's macroglobulinemia), non-IgM secreting lymphoplasmacytic lymphoma)), diffuse large B cell lymphoma (e.g., activated B In some embodiments, the subject being treated has previously received ibrutinib treatment. In some embodiments, the subject being treated has developed resistance to ibrutinib.

[0142] In some embodiments, the methods provided include inhibiting LYN and / or SRC, comprising administering to a subject a therapeutically effective amount of a compound (I) or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, isotopically labeled derivative, stereoisomer, or prodrug thereof. In some embodiments, the method further comprises administering an agent that inhibits LYN and / or SRC. For example, in some embodiments, a compound (I) described herein is administered to a subject in combination (e.g., simultaneously or sequentially) with an agent that blocks ATP binding to SRC. In some embodiments, a compound (I) described herein is administered to a subject in combination (e.g., simultaneously or sequentially) with an agent that blocks ATP binding to LYN. In some embodiments, a compound (I) described herein is administered to a subject in combination (e.g., simultaneously or sequentially) with an agent that blocks ATP binding to HCK.

[0143] In some embodiments, the method provided comprises inhibiting Tec family kinases, comprising administering to a subject a therapeutically effective amount of a compound (I) or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, isotope-labeled derivative, stereoisomer or prodrug thereof. In some embodiments, the method provided comprises inhibiting BTK, comprising administering to a subject a therapeutically effective amount of a compound (I) or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, cocrystal, isotope-labeled derivative, stereoisomer or prodrug thereof. In some embodiments, the method further comprises administering an agent that inhibits BTK and / or other Tec family kinases. For example, in some embodiments, the compound (I) described herein is administered to a subject in combination with an agent that blocks ATP from binding to Tec family kinases (e.g., simultaneously or sequentially). In some embodiments, the compound (I) described herein is administered to a subject in combination with an agent that blocks ATP from binding to BTK (e.g., simultaneously or sequentially).

[0144] In some embodiments, the selective inhibition of HCK and / or LYN and / or SRC by the compound can be determined by natural protein kinase activity profiling (e.g., KiNativ TMAs described in the Examples, in some embodiments, the ability of an inhibitor to protect a kinase from subsequent labeling with a reactive ATP-biotin probe can be determined. Living cells can be treated with compound (I), followed by cleavage treatment with ATP-biotin, and Western blotting of BTK and HCK and / or LYN and / or SRC. It can be determined whether the tested compound blocks the binding of ATP to HCK and / or LYN and / or SRC at one or more specific concentrations, and whether the tested compound blocks the binding of ATP to BTK at one or more concentrations. In such an assay, when performed under the conditions described in Example 1, blocking of binding to ATP can be determined by the lack of a detectable band on a Western blot. In some embodiments, the concentration at which the compound blocks the binding of ATP to HCK and / or LYN and / or SRC is at least 10-fold, at least 30-fold, at least 50-fold, at least 100-fold, at least 150-fold, at least 200-fold, at least 250-fold, at least 300-fold, at least 350-fold, at least 400-fold, at least 450-fold, at least 500-fold, at least 750-fold, or at least 1000-fold lower than the concentration at which it blocks the binding of ATP to BTK under equivalent conditions. In some embodiments, the concentration at which the compound blocks the binding of ATP to HCK and / or LYN and / or SRC is at least 10-fold lower than the concentration at which it blocks the binding of ATP to Bruton's tyrosine kinase (BTK) under equivalent conditions.

[0145] Those skilled in the art will appreciate that many suitable methods can be used to detect mutations in the MYD88 gene, in addition to and including the methods discussed in the examples. Detection methods that can be used include but are not limited to direct sequencing, DNA chip technology, mass spectrometry, polymerase chain reaction (PCR), allele-specific polymerase chain reaction, real-time polymerase chain reaction, reverse transcriptase PCR, electrophoretic mobility, nucleic acid hybridization, fluorescence in situ hybridization, and denaturing high performance liquid chromatography. In some embodiments, mutations in the MYD88 gene can be detected by allele-specific polymerase chain reaction (AS-PCR), for example, as described in WO 2013 / 006443.

[0146] One or more symptoms or clinical features of LPL include anemia, hyperviscosity, neuropathy, coagulopathy, splenomegaly, hepatomegaly, lymphadenopathy, and IgM serum paraprotein. In addition, subjects may also exhibit one or more of the following clinical features or symptoms of other B-cell neoplasms: asymptomatic local or systemic peripheral lymphadenopathy, plasma cell differentials, bone marrow involvement, autoimmune thrombocytopenia, peripheral blood villous lymphocytes, end-organ damage (hypercalcemia, renal insufficiency, bone lesions), recurrent infections, elevated creatine, hyperuricemia, and hypoalbuminemia. Subjects suspected of having one or more of Waldenstrom's macroglobulinemia (i.e., IgM-secreting lymphoplasmacytic lymphoma), non-IgM-secreting lymphoplasmacytic lymphoma, ABC subtype of diffuse large B-cell lymphoma, primary central nervous system (CNS) lymphoma, immune-privileged lymphoma including testicular lymphoma, marginal zone lymphoma, and chronic lymphocytic leukemia can be evaluated for the presence of mutations in the gene encoding MYD88, e.g., at position 38182641 in chromosome 3p22.2, as well as other activating mutations in MYD88 (including but not limited to V217F, W218R, I220T, S222R, M232T, S243N, and T294P).

[0147] In some embodiments, the treatment further comprises administering to the subject an agent in combination with the compounds described herein, such as an anticancer agent. In some embodiments, the treatment further comprises administering to the subject one or more of bendamustine, fludarabine, bortezomib, or idelalisib. In some embodiments, the treatment further comprises administering to the subject one or more of a BCL-2 inhibitor (e.g., venetoclax, navitoclax, obatoclax), a BCL-2 / BCL-xL inhibitor (e.g., APG-1252, BM-1197), a proteasome inhibitor (e.g., bortezomib, carfilzomib, ixazomib, or oprozomib), a monoclonal antibody (e.g., rituximab, daratumumab, ofatumumab, or obinituzumab), an alkylating agent drug (e.g., bendamustine, cyclophosphamide), a nucleoside analog (e.g., fludarabine or cladribine), an MTOR inhibitor (e.g., everolimus), a BTK inhibitor (e.g., ibrutinib, acalabrutinib, or BGB-3111), a BCR inhibitor (e.g., a SYK inhibitor), and / or an immunomodulator (e.g., thalidomide or lenalidomide). In some embodiments, the anticancer agent is a monoclonal antibody, such as rituximab. In some embodiments, the anticancer agent is a chemotherapeutic drug, such as chlorambucil, cyclophosphamide or vincristine or thalidomide. Corticosteroids, such as prednisone, may also be used in combination. Plasma exchange can be used to treat hyperviscosity syndrome by removing paraproteins from the blood. Autologous bone marrow transplantation can be used in combination with the compounds described herein. In some embodiments, the treatment further comprises administering to the subject an agent that inhibits LYN and / or SRC.

[0148] When administered to a subject, the effective amount of the therapeutic agent will depend on the specific disease being treated; the severity of the disease; individual patient parameters, including age, physical condition, size and weight, concurrent treatments, frequency of treatment, and mode of administration. These factors are well known to those of ordinary skill in the art and can be addressed only by routine experimentation. In some embodiments, a maximum dose is used, i.e., the highest safe dose based on sound medical judgment.

[0149] An effective amount of the compound will typically vary from about 0.001 mg / kg to about 1000 mg / kg, administered in one or more doses over one or more days (depending on the course of the mode of administration and the factors mentioned above).

[0150] The actual dosage level of the therapeutic agent can be varied to obtain an amount that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration. The selected dosage level depends on the activity of the particular compound, the route of administration, the tissue being treated, and the previous medical history of the patient being treated. However, it is within the skill of the art to start the dosage of the compound at a level below that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

[0151] In the treatment of MYD88 mutation diseases such as Waldenstrom's macroglobulinemia (IgM-secreting lymphoplasmacytic lymphoma), non-IgM-secreting lymphoplasmacytic lymphoma, ABC subtype of diffuse large B-cell lymphoma, primary central nervous system (CNS) lymphoma, immune privileged lymphomas including testicular lymphoma, marginal zone lymphoma, and chronic lymphocytic leukemia, an effective amount of a selective HCK inhibitor is an amount that slows disease progression, stops disease progression, or reverses disease progression. An effective amount includes, but is not limited to, an amount necessary to slow, reduce, inhibit, improve, or reverse one or more symptoms associated with a MYD88 mutation disease. In some embodiments, these terms refer to reduced levels of IgM serum paraprotein, anemia, hyperviscosity, neuropathy, coagulopathy, splenomegaly, hepatomegaly, and lymphadenopathy.

[0152] Pharmaceutical preparations and compounds are applied to subjects by any suitable route. For example, the composition can be administered orally, including sublingual, rectal, parenteral, intracisternal, intravaginal, intraperitoneal, topical and transdermal (such as by powder, ointment or drops), oral or nasal. The pharmaceutical preparations of the present disclosure may be included or diluted in a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" as used herein refers to one or more compatible fillers, diluents or other such substances, which are suitable for application to humans or other mammals, such as dogs, cats or horses. The term "carrier" means a natural or synthetic organic or inorganic ingredient, and the active ingredient is combined with it to facilitate application. The carrier can be mixed with the preparation of the present disclosure, and mixed with each other in a manner that does not significantly impair the desired drug efficacy or stability. Carriers suitable for oral, subcutaneous, intravenous, intramuscular formulations can be found in "Remington's Pharmaceutical Sciences", Mack Publishing Company, Easton, Pa., Pennsylvania. The present disclosure also provides pharmaceutical compositions comprising a compound described herein and optionally a pharmaceutically acceptable excipient.

[0153] The dosage ranges described herein provide guidance for administering provided pharmaceutical compositions to adults. The amount administered to, for example, children or adolescents can be determined by a practicing physician or person skilled in the art and can be less than or equal to the amount administered to adults.

[0154] Compounds or compositions as described herein can be administered in combination with one or more additional agents (e.g., therapeutic and / or preventive active agents). Compounds or compositions can be administered in combination with additional agents, which can improve their activity (e.g., activity (e.g., efficacy and / or efficacy)) in treating a subject in need of a disease, preventing a subject in need of a disease, inhibiting a subject in need of a kinase (e.g., SFK (e.g., LYN, HCK), Tec family kinase (e.g., BTK)) activity, improving bioavailability, improving safety, reducing drug resistance, reducing and / or changing metabolism, inhibiting excretion and / or changing distribution in a subject. It should also be understood that the therapy used can achieve the desired effect on the same illness, and / or can achieve different effects. In certain embodiments, the pharmaceutical composition comprising a compound as described herein and an additional agent as described herein shows a synergistic effect that is not present in a pharmaceutical composition comprising one of the compound and the additional agent but not both.

[0155] Compound or composition can be used with one or more other medicaments simultaneously, before or after, this can be used as, for example, conjoint therapy.Medicament comprises therapeutic active agent.Medicament also comprises preventive active agent.Medicament comprises organic small molecule, such as pharmaceutical compound (for example, FDA approved compound for human or veterinary use, as specified in federal regulations (CFR)), peptide, protein, carbohydrate, monosaccharide, oligosaccharide, polysaccharide, nucleoprotein, mucin, lipoprotein, synthetic polypeptide or protein, small molecule connected to protein, glycoprotein, steroid, nucleic acid, DNA, RNA, nucleotide, nucleoside, oligonucleotide, antisense oligonucleotide, lipid, hormone, vitamin and cell.In certain embodiments, other medicament is the medicament that can be used for treating and / or preventing disease (for example, proliferative disease, cancer, inflammatory disease, autoimmune disease, hereditary disease, blood disease, neurological disease, pain condition, mental disorder or metabolic condition) or pre-deterioration condition.Each other medicament can be used with the dosage and / or schedule determined for the medicament. Additional agents may also be administered together with each other and / or with the compounds or compositions described herein in a single dose or separately in different doses. The specific combination used in the protocol will take into account the compatibility of the compounds described herein with the additional agents and / or the desired therapeutic and / or preventive effects to be achieved. Generally, it is expected that the level of the additional agents used in combination will not exceed the level at which they are used alone. In some embodiments, the level used in combination will be lower than the level used alone.

[0156] Other agents include but are not limited to cytotoxic chemotherapeutics, epigenetic modifiers, glucocorticoids, immunotherapeutics, antiproliferative agents, anticancer agents, antiangiogenic agents, anti-inflammatory agents, immunosuppressants, antibacterial agents, antiviral agents, cardiovascular agents, cholesterol-lowering agents, antidiabetic agents, antiallergic agents, contraceptives, analgesics and combinations thereof. In certain embodiments, other agents are antiproliferative agents (e.g., anticancer agents). In certain embodiments, other agents are anti-leukemia agents. In certain embodiments, the additional agent is ABITREXATE (methotrexate), ADE, doxorubicin RDF (doxorubicin hydrochloride), Ambochlorin (chlorambucil), ARRANON (nelarabine), ARZERRA (ofatumumab), BOSULIF (bosutinib), BUSULFEX (busulfan), CAMPATH (alemtuzumab), CERUBIDINE (daunorubicin hydrochloride), CLAFEN (cyclophosphamide), CLOFAREX (clofarabine), CLOLAR (clofarabine), CVP, CYTOSAR-U (cytarabine), CYTOXAN (cyclophosphamide), ERWINAZE (asparaginase Erwinia chrysanthemi), FLUDARA (fludarabine phosphate), FOLEX (methotrexate), FOLEX PFS (methotrexate), GAZYVA (obentuzumab), GLEEVEC (imatinib mesylate), Hyper-CVAD, ICLUSIG (ponatinib hydrochloride), IMBRUVICA (ibrutinib), LEUKERAN (chlorambucil), LINFOLIZIN (chlorambucil), MARQIBO (liposomal vincristine sulfate), METHOTREXATE LPF (methotrexate), MEXATE (methotrexate), MEXATE-AQ (methotrexate), mitoxantrone hydrochloride, MUSTARGEN (mechlorethamine hydrochloride), MYLERAN (busulfan), NEOSAR (cyclophosphamide), ONCASPAR (pegaspargase), PURINETHOL (mercaptopurine), PURIXAN (mercaptopurine), daunorubicin (daunorubicin hydrochloride), SPRYCEL (dasatinib), SYNRIBO (homoharringtonine), TARABINE PFS (cytarabine), TASIGNA (nilotinib), TREANDA (bendamustine hydrochloride), TRISENOX (arsenic trioxide), VINCASAR PFS (vincristine sulfate), ZYDELIG (idelalisib), or a combination thereof. In certain embodiments, the additional agent is an anti-lymphoma agent. In certain embodiments, the additional agent is ABITREXATE (methotrexate), ABVD, ABVE, ABVE-PC, ADCETRIS (bentuximab), ADRIAMYCIN PFS (doxorubicin hydrochloride), ADRIAMYCINRDF (doxorubicin hydrochloride), AMBOCHLORIN (chlorambucil), AMBOCLORIN (chlorambucil), ARRANON (nelarabine), BEACOPP, BECENUM (carmustine), BELEODAQ (belinostat), BEXXAR (tositumomab and iodine I 131 tositumomab), BICNU (carmustine), BLENOXANE (bleomycin), CARMUBRIS (carmustine), CHOP, CLAFEN (cyclophosphamide), COPP, COPP-ABV, CVP, CYTOXAN (cyclophosphamide), DEPOCYT (liposomal cytarabine), DTIC-DOME (dacarbazine), EPOCH, FOLEX (methotrexate), FOLEX PFS (methotrexate), FOLOTYN (pralatrexate), HYPER-CVAD, ICE, IMBRUVICA (ibrutinib), INTRON A (recombinant interferon alfa-2b), ISTODAX (romidepsin), LEUKERAN (chlorambucil), LINFOLIZIN (chlorambucil), cyclohexyl lomustine, MATULANE (procarbazine hydrochloride), METHOTREXATE LPF (methotrexate), MEXATE (methotrexate), MEXATE-AQ (methotrexate), MOPP, MOZOBIL (plerixafor), MUSTARGEN (mechlorethamine hydrochloride), NEOSAR (cyclophosphamide), OEPA, ONTAK (denileukin), OPPA, R-CHOP, REVLIMID (lenalidomide), RITUXAN (rituximab), STANFORD V, TREANDA (bendamustine hydrochloride), VAMP, VELBAN (vinblastine sulfate), VELCADE (bortezomib), VELSAR (vinblastine sulfate), VINCASAR PFS (vincristine sulfate), ZEVALIN (ibritumomab tiuxetan), ZOLINZA (vorinostat), ZYDELIG (idelalisib), or a combination thereof. In certain embodiments, the additional agent is REVLIMID (lenalidomide), DACOGEN (decitabine), VIDAZA (azacitidine), CYTOSAR-U (cytarabine), IDAMYCIN (idarubicin), CERUBIDINE (daunorubicin), LEUKERAN (chlorambucil), NEOSAR (cyclophosphamide), FLUDARA (fludarabine), LEUSTATIN (cladribine), or a combination thereof. In certain embodiments, the additional agent is ABITREXATE (methotrexate), ABRAXANE (paclitaxel albumin-stabilized nanoparticle formulation), AC, AC-T, ADE, ADRIAMYCINPFS (doxorubicin hydrochloride), ADRUCIL (fluorouracil), AFINITOR (everolimus), AFINITOR DISPERZ (everolimus), ALDARA (imiquimod), ALIMTA (pemetrexed disodium), AREDIA (pamidronate disodium), ARIMIDEX (anastrozole), AROMASIN (exemestane), AVASTIN (bevacizumab), BECENUM (carmustine), BEP, BICNU (carmustine), BLENOXANE (bleomycin), CAF, CAMPTOSAR (irinotecan hydrochloride), CAPOX, CAPRELSA (vandetanib), CARBOPLATIN-TAXOL, CARMUBRIS (carmustine), CASODEX (bicalutamide), CEENU (cyclohexyl lomustine), CERUBIDINE (daunorubicin hydrochloride), CERVARIX (recombinant HPV bivalent vaccine), CLAFEN (cyclophosphamide), CMF, COMETRIQ (cabozantinib malate), COSMEGEN (dactinomycin), CYFOS (ifosfamide), CYRAMZA (ramucirumab), CYTOSAR-U (cytarabine), CYTOXAN (cyclophosphamide), DACOGEN (decitabine), DEGARELIX, DOXIL (liposomal doxorubicin hydrochloride), DOXORUBICIN HYDROCHLORIDE, DOX-SL (liposomal doxorubicin hydrochloride), DTIC-DOME (dacarbazine), EFUDEX (fluorouracil), ELLENCE (epirerubicin hydrochloride), ELOXATIN (oxaliplatin), ERBITUX (cetuximab), ERIVEDGE (vismodegib), ETOPOPHOS (etoposide phosphate), EVACET (liposomal doxorubicin hydrochloride), FARESTON (toremifene), FASLODEX (fulvestrant), FEC, FEMARA (letrozole), FLUOROPLEX (fluorouracil), FOLEX (methotrexate), PFS (methotrexate), FOLFIRI, FOLFIRI-BEVACIZUMAB, FOLFIRI-CETUXIMAB, FOLFIRINOX, FOLFOX, FU-LV, GARDASIL (recombinant human papillomavirus (HPV) quadrivalent vaccine), GEMCITABINE-CISPLATIN, GEMCITABINE-OXALIPLATIN, GEMZAR (gemcitabine hydrochloride), GILOTRIF (afatinib maleate), GLEEVEC (imatinib mesylate), GLIADEL (carmustine implant), GLIADELWAFER (carmustine implant), HERCEPTIN (trastuzumab), HYCAMTIN (topotecan hydrochloride), IFEX (ifosfamide), IFOSFAMIDUM (ifosfamide), INLYTA (axitinib), INTRON A (recombinant interferon alpha-2b), IRESSA (gefitinib), IXEMPRA (ixabepilone), JAKAFI (ruxolitinib phosphate), JEVTANA (cabazitaxel), KADCYLA (trastuzumab antibody-drug conjugate), KEYTRUDA (pembrolizumab), KYPROLIS (carfilzomib), LIPODOX (liposomal doxorubicin hydrochloride), LUPRON (leuprolide acetate), LUPRON DEPOT (leuprolide acetate), LUPRON DEPOT-3MONTH (leuprolide acetate), LUPRON DEPOT-4MONTH (leuprolide acetate), LUPRON DEPOT-PED (leuprolide acetate), MEGACE (megestrol acetate), MEKINIST (trametinib), METHAZOLASTONE (temozolomide), METHOTREXATELPF (methotrexate), MEXATE (methotrexate), MEXATE-AQ (methotrexate), MITOXANTRONEHYDROCHLORIDE, MITOZYTREX (mitomycin c), MOZOBIL (plerixafor), MUSTARGEN (mechlorethamine hydrochloride), MUTAMYCIN (mitomycin c), MYLOSAR (azacitidine), NAVELBINE (vinorelbine tartrate), NEOSAR (cyclophosphamide), NEXAVAR (sorafenib tosylate), NOLVADEX (tamoxifen citrate), NOVALDEX (tamoxifen citrate), OFF, PAD, PARAPLAT (carboplatin), PARAPLATIN (carboplatin), PEG-INTRON (pegylated interferon alpha-2b), PE METREXEDDISODIUM, PERJETA (pertuzumab), PLATINOL (cisplatin), PLATINOL-AQ (cisplatin), POMALYST (pomalidomide), prednisone, PROLEUKIN (aldesleukin), PROLIA (denosumab), PROVENGE (sipuleucel-t), REVLIMID (lenalidomide), RUBIDOMYCIN (daunorubicin hydrochloride), SPRYCEL (dasatinib), STIVARGA (regrafenib), SUTENT (sunitinib malate), SYLATRON (peginterferon alfa-2b), SYLVANT (siltuximab), SYNOVIR (thalidomide), TAC, TAFINLAR (dabrafenib), TARABINE PFS (cytarabine), TARCEVA (erlotinib hydrochloride), TASIGNA (nilotinib), TAXOL (paclitaxel), TAXOTERE (docetaxel), TEMODAR (temozolomide), THALOMID (thalidomide), TOPOSAR (etoposide), TORISEL (temsirolimus), TPF, TRISENOX (arsenic trioxide), TYKERB (lapatinib ditosylate), VECTIBIX (panitumumab), VEIP, VELBAN (vinblastine sulfate), VELCADE (bortezomib), VELSAR (vinblastine sulfate), VEPESID (etoposide), VIADUR (leuprolide acetate), VIDAZA (azacitidine), VINCASARPFS (vincristine sulfate), VOTRIENT (pazopanib hydrochloride), WELLCOVORIN (leucovorin), XALKORI (crizotinib), XELODA (capecitabine), XELOX, XGEVA (denosumab), XOFIGO (radium 223 dichloride), XTANDI (enzalutamide), YERVOY (ipilimumab), ZALTRAP (aflibercept), ZELBORAF (vemurafenib), ZOLADEX (relin acetate), ZOMETA (zoledronic acid), ZYKADIA (ceritinib), ZYTIGA (abiraterone acetate), ENMD-2076, PCI-32765, AC220, dovitinib lactate (TKI258, CHIR-258), BIBW 2992(TOVOKTM), SGX523, PF-04217903, PF-02341066, PF-299804, BMS-777607, ABT-869, MP470, BIBF 1120 AP24534, JNJ-26483327, MGCD265, DCC-2036, BMS-690154, CEP-11981, tivozanib (AV-951), OSI-930, MM-121, XL-184, XL-647 and / or XL228), proteasome inhibitors (e.g., bortezomib (Velcade)), mTOR inhibitors (e.g., rapamycin, sirolimus (CCI-779), everolimus (RAD-001), rifolimus, AP23573 (Ariad), AZD8055, BEZ235, BGT226, XL765, PF-4691502, GDC0980, SF1126 and OSI-027), oblimersen, gemcitabine, carminomycin, folinic acid, pemetrexed, cyclophosphamide, dacarbazine, procarbazine, prednisolone, dexamethasone, camptothecin, plicamycin, asparaginase, aminopterin, methotrexate, porfiromycin, melphalan, isovinblastine, vinblastine, chlorambucil, trabectedin, procarbazine, spondolactone, carminomycin, aminopterin and hexamethonium, or a combination thereof. In certain implementations, the additional agent is a cytotoxic chemotherapeutic agent (e.g., gemcitabine, cytarabine, daunorubicin, doxorubicin, vincristine, L-asparaginase, cyclophosphamide or etoposide). In certain embodiments, the additional agent is an epigenetic modifier, such as azacitidine or romidepsin. In certain embodiments, the additional agent is ruxolitinib, BBT594, CHZ868, CYT387 or BMS911543. In certain embodiments, the additional agent is an inhibitor of tyrosine kinase. In certain embodiments, the additional agent is a topoisomerase inhibitor, an MCL1 inhibitor, a BCL-2 inhibitor, a BCL-xL inhibitor, a BRD4 inhibitor, a BRCA1 inhibitor, a BRCA2 inhibitor, a HER1 inhibitor, a HER2 inhibitor, a CDK9 inhibitor, a Jumonji histone demethylase inhibitor or a DNA damage inducer. In some embodiments, the additional agent is etoposide, obatoclax, navitoclax, JQ1, 4-(((5'-chloro-2'-(((1R,4R)-4-(((R)-1-methoxypropyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridyl]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile, JIB04, or cisplatin. In certain embodiments, the additional agent is a binder or inhibitor of a kinase (e.g., a SRC family kinase (e.g., HCK, LYN, BLK, FRK), a Tec family kinase (e.g., BTK)). In certain embodiments, the additional agent is an antibody or fragment thereof (e.g., a monoclonal antibody). In certain embodiments, the additional agent is a tyrosine kinase inhibitor.In certain embodiments, the additional agent is selected from the group consisting of: epigenetic or transcriptional regulators (e.g., DNA methyltransferase inhibitors, histone deacetylase inhibitors (HDAC inhibitors), lysine methyltransferase inhibitors), anti-mitotic drugs (e.g., taxanes and vinca alkaloids), hormone receptor modulators (e.g., estrogen receptor modulators and androgen receptor modulators), cell signaling pathway inhibitors (e.g., tyrosine protein kinase inhibitors), protein stability regulators (e.g., proteasome inhibitors), Hsp90 inhibitors, glucocorticoids, all-trans retinoic acid and other agents that promote differentiation. In certain embodiments, the additional agent is a glucocorticoid (e.g., cortisol, cortisone, prednisone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, fludrocortisone acetate or deoxycorticosterone acetate). In certain embodiments, the additional therapy is an immunotherapy (e.g., an immunotherapeutic monoclonal antibody). In certain embodiments, the additional agent is an immunomodulator. In certain embodiments, the additional agent is an immune checkpoint inhibitor. In certain embodiments, the additional agent is a programmed cell death 1 protein (PD-1) inhibitor. In certain embodiments, the additional agent is a programmed cell death 1 protein ligand 1 (PD-L1) inhibitor. In certain embodiments, the additional agent is a cytotoxic T lymphocyte-associated protein 4 (CTLA-4) inhibitor. In certain embodiments, the additional agent is a T cell immunoglobulin domain and mucin domain 3 (TIM3) inhibitor, a lymphocyte activation gene-3 (LAG3) inhibitor, a T cell activation inhibitor 1 (VTCN1 or B7-H4) inhibitor containing a V-set domain, a differentiation cluster 276 (CD276 or B7-H3) inhibitor, a B and T lymphocyte attenuator (BTLA) inhibitor, a galectin-9 (GAL9) inhibitor, a checkpoint kinase 1 (Chk1) inhibitor, adenosine A2A receptor (A2AR) inhibitor, an indoleamine 2,3-dioxygenase (IDO) inhibitor, a killer cell immunoglobulin-like receptor (KIR) inhibitor, or a V domain Ig inhibitor of T cell activation (VISTA) inhibitor. In certain embodiments, the PD-1 inhibitor is nivolumab, pidilizumab, pembrolizumab, MEDI-0680, REGN2810, or AMP-224. In certain embodiments, the PD-L1 inhibitor is atezolizumab, durvalumab, BMS-936559, avelumab, or CA- 170. In certain embodiments, the CTLA-4 inhibitor is ipilimumab or tremelimumab.In certain embodiments, the compounds or pharmaceutical compositions described herein may be administered in combination with anti-cancer therapy, including but not limited to surgery, radiation therapy, and transplantation (eg, stem cell transplantation, bone marrow transplantation).

[0157] In certain embodiments, the additional agent is a BCL-2 inhibitor (eg, venetoclax, navitoclax, obatoclax), a BCL-2 / BCL-xL inhibitor (eg, APG-1252, BM-1197).

[0158] In certain embodiments, the additional agent is venetoclax.

[0159] In certain embodiments, the compounds described herein are provided in an effective amount (e.g., effective to inhibit a kinase, such as a SRC family kinase (e.g., HCK, LYN, BLK, FRK) or a Tec family kinase (e.g., BTK)) in a pharmaceutical composition. In certain embodiments, the effective amount is a therapeutically effective amount. In certain embodiments, the therapeutically effective amount is an amount that effectively inhibits a kinase. In certain embodiments, the therapeutically effective amount is an amount that effectively treats a disease (e.g., a disease associated with abnormal activity of a kinase (e.g., a proliferative disease)). In certain embodiments, the therapeutically effective amount is an amount that effectively inhibits kinase activity and treats a disease (e.g., a disease associated with abnormal activity of a kinase (e.g., a proliferative disease)). In certain embodiments, the therapeutically effective amount is an amount that effectively induces apoptosis in a cell (e.g., a malignant cell, a pre-malignant cell).

[0160] In certain embodiments, an effective amount is an amount effective to inhibit kinase activity by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 98%. In certain embodiments, an effective amount is an amount effective to inhibit kinase activity by no more than 10%, no more than 20%, no more than 30%, no more than 40%, no more than 50%, no more than 60%, no more than 70%, no more than 80%, no more than 90%, no more than 95%, or no more than 98%.

[0161] In certain embodiments, the subject is an animal. The animal can be of either sex and can be at any stage of development. In certain embodiments, the subject described herein is a human (e.g., an adult, a teenager, or a child). In certain embodiments, the subject is a non-human animal. In certain embodiments, the subject is a mammal. In certain embodiments, the subject is a non-human mammal. In certain embodiments, the subject is a domestic animal, such as a dog, a cat, a cow, a pig, a horse, a sheep, or a goat. In certain embodiments, the subject is a dog. In certain embodiments, the subject is a companion animal, such as a dog or a cat. In certain embodiments, the subject is a livestock animal, such as a cow, a pig, a horse, a sheep, or a goat. In certain embodiments, the subject is a zoo animal. In another embodiment, the subject is a research animal, such as a rodent (e.g., a mouse, a rat), a dog, a pig, or a non-human primate. In certain embodiments, the subject is a genetically engineered animal. In certain embodiments, the subject is a transgenic animal (e.g., a transgenic mouse, a transgenic pig). In certain embodiments, the subject is a fish or a reptile.

[0162] The pharmaceutical compositions described herein can be prepared by any method known in the art of pharmacology. Generally, such preparation methods include combining the compound described herein (i.e., "active ingredient") with a carrier or excipient and / or one or more other auxiliary ingredients, and then, if necessary and / or desired, shaping and / or packaging the product into a desired single or multiple dose unit.

[0163] Pharmaceutical compositions can be prepared, packaged and / or sold in bulk as a single unit dose and / or as multiple single unit doses. A "unit dose" is a discrete amount of a pharmaceutical composition comprising a predetermined amount of an active ingredient. The amount of the active ingredient is generally equal to the dose of the active ingredient administered to a subject and / or a convenient fraction of such a dose, such as one-half or one-third of the dose.

[0164] The relative amounts of the active ingredient, pharmaceutically acceptable excipient and / or any additional ingredients in the pharmaceutical compositions described herein will vary depending on the identity, size and / or condition of the subject being treated and further on the route of administration of the composition. The composition may include 0.1% to 100% (w / w) of the active ingredient.

[0165] Pharmaceutically acceptable excipients for preparing the provided pharmaceutical compositions include inert diluents, dispersants and / or granulating agents, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants and / or oils. Excipients such as cocoa butter and suppository waxes, colorants, coating agents, sweeteners, flavoring agents and fragrances may also be present in the compositions.

[0166] Exemplary diluents include calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, dibasic calcium phosphate, sodium phosphate lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and mixtures thereof.

[0167] Exemplary granulating and / or dispersing agents include potato starch, corn starch, tapioca starch, sodium starch glycolate, clays, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose and wood products, natural sponges, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinyl pyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (croscarmellose), methyl cellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and mixtures thereof.

[0168] Exemplary surfactants and / or emulsifiers include natural emulsifiers (e.g., gum arabic, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan gum, pectin, gelatin, egg yolk, casein, lanolin, cholesterol, waxes, and lecithin), colloidal clays (e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminum silicate)), long chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, glyceryl monostearate, ethylene glycol distearate, glyceryl monostearate and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxypolymethylene, polyacrylic acid, acrylic acid polymers, and carboxyvinyl polymers), carrageenan, cellulose derivatives (e.g., sodium carboxymethylcellulose, powdered cellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate ( 20) Polyoxyethylene sorbitan ( 60), polyoxyethylene sorbitan monooleate ( 80), Sorbitan monopalmitate ( 40) Sorbitan monostearate ( 60), Sorbitan tristearate ( 65), glyceryl monooleate, sorbitan monooleate ( 80), polyoxyethylene esters (e.g., polyoxyethylene monostearate ( 45), polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate and ), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g., ), polyoxyethylene ethers (e.g., polyoxyethylene lauryl ether ( 30)), poly(vinyl pyrrolidone), diethylene glycol laurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, F-68, poloxamer P-188, cetyltrimethylammonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium and / or mixtures thereof.

[0169] Exemplary binders include starches (e.g., corn starch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextran, dextrin, molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums (e.g., gum arabic, sodium alginate, extract of carrageenan, panwar gum, gum ghatti, mucilage of esophageal shell, carboxymethyl cellulose, methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, microcrystalline cellulose, cellulose acetate, poly(vinyl pyrrolidone), magnesium aluminum silicate, and larch arabinogalactan), alginate, polyethylene oxide, polyethylene glycol, inorganic calcium salt, silicic acid, polymethacrylate, wax, water, alcohol and / or mixtures thereof.

[0170] Exemplary preservatives include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, antiprotozoal preservatives, alcohol preservatives, acidic preservatives and other preservatives. In certain embodiments, the preservative is an antioxidant. In other embodiments, the preservative is a chelating agent.

[0171] Exemplary antioxidants include alpha tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite.

[0172] Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA) and salts and hydrates thereof (e.g., sodium edetate, disodium edetate, trisodium edetate, calcium disodium edetate, dipotassium edetate, etc.), citric acid and salts and hydrates thereof (e.g., citric acid monohydrate), fumaric acid and salts and hydrates thereof, malic acid and salts and hydrates thereof, phosphoric acid and salts and hydrates thereof, and tartaric acid and salts and hydrates thereof. Exemplary antimicrobial preservatives include benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethanol, glycerol, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and thimerosal.

[0173] Exemplary antifungal preservatives include butylparaben, methylparaben, ethylparaben, propylparaben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid.

[0174] Exemplary alcohol preservatives include ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoates, and phenethyl alcohol.

[0175] Exemplary acidic preservatives include vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.

[0176] Other preservatives include tocopherol, tocopherol acetate, mesylate, cetyltrimethylammonium bromide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, Plus, Methylparaben, 115. II. and

[0177] Exemplary buffers include citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glucuronate, calcium glucoheptonate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propionic acid, calcium levulinate, valeric acid, calcium hydrogen phosphate, phosphoric acid, calcium orthophosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dipotassium hydrogen phosphate, monopotassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, disodium hydrogen phosphate, monosodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethanol, and mixtures thereof.

[0178] Exemplary lubricants include magnesium stearate, calcium stearate, stearic acid, silicon dioxide, talc, malt, glyceryl behenate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and mixtures thereof.

[0179] Exemplary natural oils include almond, bitter almond, avocado, babassu, bergamot, black flower seed, borage, juniper berry, chamomile, canola, caraway, castor, cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cotton seed, emu, eucalyptus, evening primrose, fish, flax seed, geraniol, gourd, grape seed, hazelnut, hyssop, isopropyl myristate, jojoba, macadamia, lavender, lavender essence. Oil, lemon, litsea cubeba, macadamia nut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange, orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed, pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood, sasquana, savory, sea buckthorn, sesame, shea butter, silicone, soybean, sunflower, tea tree, thistle, toon, vetiver, walnut and wheat germ oil. Exemplary synthetic oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil and mixtures thereof.

[0180] Liquid dosage forms for oral and parenteral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage form may include inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan, and mixtures thereof. In addition to inert diluents, oral compositions may include adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and aromatics. In certain embodiments for parenteral administration, the conjugates described herein are mixed with a solubilizer (e.g. alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers and mixtures thereof).

[0181] Injectable preparations, such as sterile injectable aqueous or oily suspensions, can be prepared according to known techniques using suitable dispersants or wetting agents and suspending agents. Sterile injectable preparations can be sterile injectable solutions, suspensions or emulsions in nontoxic parenteral acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable carriers and solvents that can be used are water, Ringer's solution, USP and isotonic sodium chloride solution. In addition, sterile fixed oils are generally used as solvents or suspension media. For this purpose, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid are used to prepare injections.

[0182] The injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0183] In order to prolong the effect of the drug, it is usually necessary to slow down the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of a crystalline or amorphous material with poor water solubility. The absorption rate of the drug depends on its dissolution rate, which in turn can depend on crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form can be achieved by dissolving or suspending the drug in an oil vehicle.

[0184] Compositions for rectal or vaginal administration are typically suppositories, which can be prepared by mixing the conjugates described herein with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax, which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active ingredient.

[0185] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active ingredient is mixed with at least one inert pharmaceutically acceptable excipient or carrier (such as sodium citrate or dicalcium phosphate) and / or (a) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol and silicic acid, (b) binders such as carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and acacia, (c) humectants such as glycerol, (d) disintegrants such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate, (e) solution retardants such as paraffin, (f) absorption promoters such as quaternary ammonium compounds, (g) wetting agents such as cetyl alcohol and glyceryl monostearate, (h) absorbents such as kaolin and bentonite, and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may contain a buffer.

[0186] Solid compositions of similar types can be used as fillers in soft and hard filled gelatin capsules, using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings and other coatings known in the field of pharmacology. They can optionally include opacifiers, and can be compositions that release active ingredients only in or preferably in a certain part of the intestinal tract, optionally in a delayed manner. Examples of usable encapsulated compositions include polymeric substances and waxes. Solid compositions of similar types can be used as fillers in soft and hard filled gelatin capsules, using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol.

[0187] The active ingredient can be a microencapsulated form with one or more excipients as described above. The solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings, controlled release coatings and other coatings known in the field of drug formulation. In such solid dosage forms, the active ingredient can be mixed with at least one inert diluent such as sucrose, lactose or starch. As normally practiced, such dosage forms can include other substances other than inert diluents, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage form can include a buffer. They can optionally include an opacifier, and can be a composition that releases the active ingredient only in or preferably in a certain part of the intestinal tract in a delayed manner. The example of an encapsulating agent that can be used includes polymeric substances and waxes.

[0188] The dosage form for topical and / or transdermal administration of the compounds described herein may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants and / or patches. Typically, the active ingredient is mixed with a pharmaceutically acceptable carrier or excipient and / or any required preservative and / or buffer that may be required under aseptic conditions. In addition, the disclosure contemplates the use of transdermal patches, which generally have the additional advantage of providing controlled delivery of active ingredients to the body. Such dosage forms can be prepared, for example, by dissolving and / or dispersing the active ingredient in a suitable medium. Alternatively or additionally, the rate can be controlled by providing a rate-controlled membrane and / or by dispersing the active ingredient in a polymer matrix and / or a gel.

[0189] Suitable devices for delivering intradermal pharmaceutical compositions described herein include short needle devices. Intradermal compositions can be applied by devices that limit the effective length of needle penetration into the skin. Alternatively or additionally, conventional syringes can be used in the classical Mantoux method of intradermal administration. It is suitable to deliver liquid formulations to the dermis by a liquid jet injector and / or by a needle that pierces the stratum corneum and produces a jet that reaches the dermis. It is suitable to use a ballistic powder / particle delivery device that accelerates a compound in powder form through the outer layer of the skin to reach the dermis using compressed gas.

[0190] Preparations suitable for topical administration include, but are not limited to, liquid and / or semi-liquid preparations, such as liniments, lotions, oil-in-water and / or water-in-oil emulsions, such as creams, ointments and / or pastes, and / or solutions and / or suspensions. Topically administrable preparations may, for example, include about 1% to about 10% (w / w) of the active ingredient, although the concentration of the active ingredient may be as high as the solubility limit of the active ingredient in the solvent. Preparations for topical administration may further include one or more additional ingredients as described herein.

[0191] The pharmaceutical compositions described herein can be prepared, packaged and / or sold in a formulation suitable for oral administration through the lungs. Such formulations can include dry particles including active ingredients and having a diameter in the range of about 0.5 to about 7 nanometers, or about 1 to about 6 nanometers. Such compositions are conveniently in the form of dry powders for use with a device including a dry powder reservoir and / or using a self-propelling solvent / powder dispensing container (such as a device including an active ingredient dissolved and / or suspended in a low-boiling propellant in a sealed container), and the propellant stream can be directed to the dry powder reservoir to disperse the powder. Such powders include particles, wherein at least 98% by weight of the particles have a diameter greater than 0.5 nanometers, and at least 95% by number of the particles have a diameter less than 7 nanometers. Alternatively, at least 95% by weight of the particles have a diameter greater than 1 nanometer, and at least 90% by number of the particles have a diameter less than 6 nanometers. The dry powder composition can include a solid fine powder diluent, such as sugar, and is conveniently provided in a unit dosage form.

[0192] Low boiling point propellants typically comprise liquid propellants having a boiling point below 65° F. at atmospheric pressure. Typically, the propellant may constitute 50 to 99.9% (w / w) of the composition, while the active ingredient may constitute 0.1 to 20% (w / w) of the composition. The propellant may further comprise additional ingredients, such as liquid nonionic and / or solid anionic surfactants and / or solid diluents (which may have a particle size of the same order of magnitude as the particles comprising the active ingredient).

[0193] The pharmaceutical compositions described herein formulated for pulmonary delivery can provide active ingredients in the form of droplets of solutions and / or suspensions. Such preparations can be prepared, packaged and / or sold as optionally sterile aqueous and / or diluted alcohol solutions and / or suspensions comprising active ingredients, and can be conveniently applied using any spray and / or atomizing device. Such preparations can further include one or more additional ingredients, including but not limited to flavorings such as saccharin sodium, volatile oils, buffers, surfactants and / or preservatives such as methyl hydroxybenzoate. The droplets provided by this route of administration can have an average diameter of about 0.1 to about 200 nanometers.

[0194] The formulations described herein that can be used for pulmonary delivery can be used for intranasal delivery of the pharmaceutical compositions described herein. Another formulation suitable for intranasal administration is a coarse powder comprising the active ingredient and having an average particle size of about 0.2 to 500 microns. This formulation is administered by rapid inhalation through the nasal passages from a powder container held near the nostrils.

[0195] Preparations for nasal administration may, for example, include as little as about 0.1% (w / w) to as much as 100% (w / w) of active ingredients, and may include one or more additional ingredients described herein. The pharmaceutical compositions described herein may be prepared, packaged and / or sold in preparations for oral administration. Such preparations may be in the form of tablets and / or lozenges, for example, prepared using conventional methods, and may contain, for example, 0.1 to 20% (w / w) of active ingredients, the remainder including orally soluble and / or degradable compositions and optionally one or more additional ingredients described herein. Alternatively, preparations for oral administration may include powders and / or aerosolized and / or atomized solutions and / or suspensions including active ingredients. When dispersed, such powdered, aerosolized and / or aerosolized preparations may have an average particle and / or droplet size of about 0.1 to about 200 nanometers, and may further include one or more additional ingredients described herein.

[0196] The pharmaceutical compositions described herein can be prepared, packaged and / or sold in the form of formulations for ocular administration. Such formulations can be, for example, in the form of eye drops, comprising, for example, a 0.1 to 1.0% (w / w) solution and / or suspension of the active ingredient in an aqueous or oily liquid carrier or excipient. Such drops can further include a buffer, a salt and / or one or more additional ingredients described herein. Other useful ocularly administrable formulations include those comprising active ingredients in microcrystalline form and / or liposomal formulations. Ear drops and / or eye drops are also encompassed within the scope of the present disclosure.

[0197] Although the descriptions of pharmaceutical compositions provided herein are primarily directed to pharmaceutical compositions suitable for administration to humans, it will be understood by those skilled in the art that such compositions are generally suitable for administration to animals of all kinds. Modifications of pharmaceutical compositions suitable for administration to humans in order to make the compositions suitable for administration to various animals are well known, and such modifications can be designed and / or performed by an ordinary skilled veterinary pharmacologist using ordinary experimentation.

[0198] The compounds provided herein are generally formulated in dosage unit forms for ease of administration and uniform dosage. However, it should be understood that the total daily dosage of the compositions described herein will be determined by the physician within the scope of reasonable medical judgment. The specific therapeutically effective dosage level for any particular subject or organism will depend on a variety of factors, including the severity of the disease and condition being treated; the activity of the specific active ingredient used; the specific composition used; the age, weight, general health, sex and diet of the subject; the time of administration, route of administration and excretion rate of the specific active ingredient used; the duration of treatment; drugs used in combination or concomitantly with the specific active ingredient used; and similar factors known in the medical field.

[0199] Compounds and compositions provided herein can be administered by any route, including enteral (e.g., oral), parenteral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, subcutaneous, intraventricular, transdermal, intradermal, rectal, intravaginal, intraperitoneal, topical (such as by powder, ointment, cream and / or drops), mucosal, nasal, oral, sublingual; by intratracheal instillation, bronchial instillation and / or inhalation; and / or as oral spray, nasal spray and / or aerosol. Specific considerations are oral administration, intravenous administration (e.g., systemic intravenous injection), regional administration via blood and / or lymph supply, and / or direct administration to the affected site. In general, the most appropriate route of administration will depend on a variety of factors, including the properties of the medicament (e.g., its stability in the gastrointestinal environment) and / or the subject's condition (e.g., whether the subject can tolerate oral administration). In certain embodiments, the compound or pharmaceutical composition described herein is suitable for topical administration to the subject's eyes.

[0200] The exact amount of compound required to achieve an effective amount will vary from subject to subject, depending on, for example, the species, age and overall condition of the subject, the severity of the side effects or obstacles, the characteristics of the specific compound, the mode of administration, etc. The effective amount can be contained in a single dose (e.g., a single oral dose) or multiple doses (e.g., multiple oral doses). In certain embodiments, when multiple doses are administered to a subject, any two doses of the multiple doses contain different or substantially the same amount of compounds as described herein. In certain embodiments, when multiple doses are administered to a subject, the frequency of administering multiple doses to the subject is three doses a day, two doses a day, one dose a day, one dose every other day, one dose every three days, one dose a week, one dose every two weeks, one dose every three weeks, or one dose every four weeks. In certain embodiments, the frequency of administering multiple doses to a subject is one dose a day. In certain embodiments, the frequency of administering multiple doses to a subject is two doses a day. In certain embodiments, the frequency of administering multiple doses to a subject is three doses a day. In certain embodiments, when multiple doses are administered to a subject, the duration between the first dose and the last dose of the multiple doses is one day, two days, four days, one week, two weeks, three weeks, one month, two months, three months, four months, six months, nine months, one year, two years, three years, four years, five years, seven years, ten years, fifteen years, twenty years, or the life of the subject or cell. In certain embodiments, the duration between the first dose and the last dose of the multiple doses is three months, six months, or one year. In certain embodiments, the duration between the first dose and the last dose of the multiple doses is the life of the subject. In certain embodiments, the dosage described herein (e.g., any dosage of a single dose or multiple doses) independently comprises 0.1 μg to 1 μg, 0.001 mg to 0.01 mg, 0.01 mg to 0.1 mg, 0.1 mg to 1 mg, 1 mg to 3 mg, 3 mg to 10 mg, 10 mg to 30 mg, 30 mg to 100 mg, 100 mg to 300 mg, 300 mg to 1,000 mg, or 1 g to 10 g (including end values) of the compounds described herein. In certain embodiments, the dosages described herein independently include 1 mg to 3 mg (including end values) of the compounds described herein. In certain embodiments, the dosages described herein independently include 3 mg to 10 mg (including end values) of the compounds described herein. In certain embodiments, the dosages described herein independently include 10 mg to 30 mg (including end values) of the compounds described herein. In certain embodiments, the dosages described herein independently include 30 mg to 100 mg (including end values) of the compounds described herein.

[0201] The present disclosure also encompasses a kit (e.g., a pharmaceutical package). In certain embodiments, the kit includes a compound (I) or a pharmaceutical composition as described herein, and instructions for using the compound or pharmaceutical composition. In certain embodiments, the kit includes a first container, wherein the first container includes a compound or pharmaceutical composition. In certain embodiments, the kit further includes a second container. In certain embodiments, the second container includes an excipient (e.g., an excipient for diluting or suspending the compound or pharmaceutical composition). In certain embodiments, the second container includes an additional medicament. In certain embodiments, the kit further includes a third container. In certain embodiments, the third container includes an additional medicament. In certain embodiments, the compound or pharmaceutical composition contained in the first container is combined with an excipient or additional medicament contained in the second container to form a unit dosage form. In certain embodiments, the compound or pharmaceutical composition contained in the first container, the excipient contained in the second container, and the additional medicament contained in the third container are combined to form a unit dosage form. In certain embodiments, the first container, the second container, and the third container are each independently a vial, an ampoule, a bottle, a syringe, a dispenser package, a tube, or an inhaler.

[0202] In certain embodiments, instructions are used to administer a compound or pharmaceutical composition to a subject (e.g., a subject in need of treating or preventing a disease described herein). In certain embodiments, instructions include information required by regulatory agencies such as the U.S. Food and Drug Administration (FDA) or the European Medicines Evaluation Agency (EMA). In certain embodiments, instructions include prescription information.

[0203] In another aspect, the present disclosure provides a method of treating a disease in a subject in need thereof, the method comprising administering to the subject in need thereof an effective amount (eg, a therapeutically effective amount) of a compound (I) described herein or a pharmaceutical composition described herein.

[0204] In another aspect, the present disclosure provides a method of preventing a disease in a subject in need thereof, the method comprising administering to the subject in need thereof an effective amount (eg, a prophylactically effective amount) of a compound described herein or a pharmaceutical composition described herein.

[0205] In another aspect, the present disclosure provides a method of inhibiting kinase activity in a subject in need thereof, the method comprising administering to the subject in need thereof an effective amount of a compound described herein or a pharmaceutical composition described herein.

[0206] Without wishing to be bound by any particular theory, in certain embodiments, the compounds described herein are capable of binding to an inhibited kinase. In certain embodiments, the compounds described herein are capable of binding to a kinase. In certain embodiments, the kinase is a SFK (e.g., HCK, LYN, BLK, FRK). In certain embodiments, the kinase is HCK. In certain embodiments, the kinase is LYN. In certain embodiments, the kinase is a Tec family kinase (e.g., BTK).

[0207] In certain embodiments, the kinase is BTK. In certain embodiments, there is provided a method of reducing the activity of a kinase (e.g., SFK (e.g., HCK, LYN, BLK, FRK), Tec family kinase (e.g., BTK)) in a subject by at least about 1%, at least about 3%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80% or at least about 90%. In certain embodiments, the activity of kinase in a subject is reduced by at least about 1%, at least about 3%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80% or at least about 90%. In some embodiments, the activity of kinase in a subject is selectively inhibited by the method. In some embodiments, the activity of kinase (e.g., HCK, LYN, BTK) in a subject is selectively reduced by a compound or pharmaceutical composition as described herein.

[0208] Diseases comprising proliferative diseases can be associated with abnormal or undesirable activity of kinases and / or overexpression of kinases. Abnormal or undesirable activity of kinases can be an increase or decrease in the level of kinase activity. Proliferative diseases are sometimes associated with abnormal levels of JAK activity, typically by increasing or decreasing JAK activation. Inhibition of JAK2 activity is expected to inhibit phosphorylation. In certain embodiments, JAK2 is not overexpressed, but the activity of JAK2 increases. In certain embodiments, JAK2 is overexpressed, and the activity of JAK2 increases. The compounds and pharmaceutical compositions described herein can inhibit the activity of JAK2 and can be used to treat and / or prevent diseases, such as diseases associated with abnormal, increased or undesirable activity of kinases, overactivation of kinases and / or overexpression of kinases.

[0209] In certain embodiments, the disease (e.g., a disease to be treated or prevented by the methods described herein) is associated with increased activity of a kinase (e.g., SFK (e.g., HCK, LYN, BLK, FRK), Tec family kinase (e.g., BTK)). In certain embodiments, the disease is associated with overexpression of a kinase (e.g., SFK (e.g., HCK, LYN, BLK, FRK), Tec family kinase (e.g., BTK)). In certain embodiments, the disease is a proliferative disease. In certain embodiments, the proliferative disease is cancer. In certain embodiments, cancer is associated with mutations in MYD88. In another embodiment, cancer is associated with mutated BTK. In certain embodiments, the proliferative disease is mastocytosis. In certain embodiments, mastocytosis is systemic mastocytosis. In certain embodiments, the proliferative disease is IgM gammopathy. In certain embodiments, the IgM gammopathy is an IgM monoclonal gammopathy of undetermined significance.

[0210] In certain embodiments, the disease is breast cancer. In certain embodiments, the disease is colon cancer. In certain embodiments, the disease is testicular cancer. In certain embodiments, the disease is CNS cancer. In certain embodiments, the disease is gastric cancer. In certain embodiments, the disease is lymphoma. In certain embodiments, lymphoma is B cell lymphoma. In certain embodiments, B cell lymphoma is lymphoplasmacytic lymphoma. In certain embodiments, lymphoplasmacytic lymphoma is lymphoplasmacytic lymphoma (i.e., Waldenstrom's macroglobulinemia) that secretes IgM. In certain embodiments, the disease is Waldenstrom's macroglobulinemia. In certain embodiments, lymphoplasmacytic lymphoma is lymphoplasmacytic lymphoma that does not secrete IgM. In certain embodiments, lymphoma is diffuse large B cell lymphoma (DLBCL). In certain embodiments, DLBCL is activated B cell-like (ABC)-DLBCL. In certain embodiments, DLBCL is germinal center B cell-like (GBC)-DLBCL. In certain embodiments, the lymphoma is follicular lymphoma. In certain embodiments, the lymphoma is marginal zone B-cell lymphoma. In certain embodiments, the lymphoma is small lymphocytic lymphoma. In certain embodiments, the small lymphocytic lymphoma is mantle cell lymphoma. In certain embodiments, the cancer is leukemia. In certain embodiments, the leukemia is chronic lymphocytic leukemia (CLL). In certain embodiments, the leukemia is myeloid leukemia. In certain embodiments, the myeloid leukemia is chronic myeloid leukemia. In certain embodiments, the myeloid leukemia is acute myeloid leukemia. In certain embodiments, the acute myeloid leukemia is mast cell leukemia. In certain embodiments, the cancer is myeloma. In certain embodiments, the myeloma is IgM myeloma. In certain embodiments, the IgM myeloma is IgM multiple myeloma. In certain embodiments, the cancer is a myeloproliferative disease. In certain embodiments, the myeloproliferative disease is myelodysplastic syndrome.

[0211] In certain embodiments, the methods described herein are superior to (e.g., show improved safety and / or therapeutic efficacy) or comparable to existing therapies (e.g., chemotherapy, treatment with BTK inhibitors). In certain embodiments, the methods described herein are associated with reduced toxicity when compared to existing therapies (e.g., chemotherapy, treatment with BTK inhibitors).

[0212] In certain embodiments, the cell is a malignant cell (e.g., a cancer cell). In certain embodiments, the cell is a malignant blood cell. In certain embodiments, the cell is a malignant bone marrow cell. In certain embodiments, the cell is an adenocarcinoma cell, a blastoma cell, a cancer cell, or a sarcoma cell. In certain embodiments, the cell is a premalignant cell (e.g., a precancerous cell).

[0213] In certain embodiments, the method described herein further includes applying other therapy to a subject in need. In certain embodiments, other therapy is cytotoxic chemotherapy (for example, gemcitabine, cytarabine, daunorubicin, doxorubicin, vincristine, L-asparaginase, cyclophosphamide or etoposide). In certain embodiments, other therapy is an epigenetic modifier (for example, azacitidine or romidepsin). In certain embodiments, other therapy is a glucocorticoid. In certain embodiments, other therapy is an immunotherapy (for example, immunotherapeutic monoclonal antibody). In certain embodiments, other medicament is etoposide, obatoclax or navitoclax, and optionally, disease is breast cancer, such as triple negative breast cancer, HER2 positive breast cancer, HER2 negative breast cancer, ER positive breast cancer, ER negative breast cancer or ER / PR positive breast cancer. In certain embodiments, other medicament is etoposide, JIB04 or cisplatin, and optionally, disease is Ewing's sarcoma. In some embodiments, the additional drug is JQ1 or NVP2, and optionally, the disease is a leukemia, such as acute myeloid leukemia, myeloblastic leukemia, promyelocytic leukemia, myelomonocytic leukemia, monocytic leukemia, monoblastic leukemia, or megakaryocytic leukemia.

[0214] In yet another aspect, the disclosure provides compounds and pharmaceutical compositions described herein for use in treating a disease (eg, a proliferative disease such as IgM gammopathy, mastocytosis, or cancer) in a subject in need thereof.

[0215] In yet another aspect, the disclosure provides compounds and pharmaceutical compositions described herein for use in preventing a disease (eg, a proliferative disease such as IgM gammopathy, mastocytosis, or cancer) in a subject in need thereof.

[0216] In another aspect, the present disclosure provides compounds and pharmaceutical compositions described herein for use in inhibiting the activity of a kinase in a subject in need thereof.

[0217] In another aspect, the present disclosure provides the use of the compounds and pharmaceutical compositions described herein in the preparation of a medicament for treating a disease in a subject in need thereof.

[0218] In another aspect, the present disclosure provides for the use of the compounds and pharmaceutical compositions described herein in the preparation of a medicament for preventing a disease in a subject in need thereof.

[0219] The compounds, pharmaceutical compositions, and kits described herein can synergistically enhance the inhibition of kinases (e.g., SFKs (e.g., HCK, LYN, BLK, FRK), TEC family kinases (e.g., BTK)) induced by additional agents in a subject. Thus, the combination of a compound, pharmaceutical composition, or kit with an additional agent can be used to treat a disease that is resistant to treatment with an additional agent without a compound, pharmaceutical composition, or kit described herein.

[0220] The present disclosure is further illustrated by the following examples, which should in no way be construed as further limiting.The entire contents of all references cited in this application (including literature references, published patents, published patent applications, and co-pending patent applications) are expressly incorporated herein by reference.

[0221] Examples

[0222] Example 1. Compound (I) selectively targets HCK and BTK in MYD88-mutated WM and ABC DLBCL cells.

[0223] Figure 2 shows the relationship between compound (I) and A419259 Kinase selectivity curves. The compounds were analyzed by DiscoverX at a concentration of 1.0 μM against a panel of over 460 kinases. Compound (I) and A419259 had nearly identical selectivity, with an S(10) score of 0.13 and an S(1) score of 0.07. Enzymatic assays were also performed on potential targets HCK, BTK, LYN (SelectScreen, Life Technologies), which demonstrated that compound (I) exhibited potent inhibition of HCK, BTK, and LYN with single-digit nanomolar IC50s, while A419259 was 10-fold less potent against BTK.

[0224] Biochemical kinase assays demonstrated that compound (I) strongly inhibited HCK (IC 50 <0.495 nM) and BTK (IC50 = 0.915 nM). In contrast, ibrutinib showed similar BTK inhibition (IC 50 =0.614 nM) and 100-fold less potent HCK inhibition (IC50 = 49 nM). To evaluate the kinase target selectivity of compound (I), a panel of 464 kinases was tested. Compound (I) showed high selectivity (S10 = 0.07; S35 = 0.13), which was comparable to the selectivity of ibrutinib (S10 = 0.04; S35 = 0.12). The kinases targeted by compound (I) were mainly limited to Src family kinases HCK, BLK, LYN and FRK, and Tec family kinases BTK ( Figure 2ATo further evaluate the kinase selectivity of compound (I) and verify its target engagement in living cells, KiNativ TM Assay that measures the ability of compound (I) to block kinase binding to a desthiobiotin-ATP probe. KiNativ TM The screening confirmed that compound (I) strongly targeted both HCK and BTK in TMD8 ABC DLBCL cells treated at 1.0 μM. These experiments showed that compound (I) also had nanomolar activity against SRC family kinases (e.g., HCK, BLK, LYN, FRK), Tec family kinases (e.g., BTK), ACK family kinases (e.g., ACK (i.e., TNK2)), CSK, ErbB2, and ABL ( Figure 2B To further confirm that compound (I) targets HCK and BTK, the Pierce ELISA with ActivX desthiobiotin-ATP probe was also used after pre-treatment of live cells with compound (I) in MYD88-mutated BCWM.1WM cells and TMD8 ABC DLBCL cells. TM The kinase enrichment kit was used to perform an ATP-competitive assay. The enriched kinases were analyzed by Western blotting and showed potent binding of compound (I) to HCK and similar levels of BTK inhibition as ibrutinib ( Figure 2C ). Compound (I) binds BTK efficiently and very strongly to HCK and LYN in living BCWM.1 and TMD8 cells.

[0225] Example 2. Inhibition of HCK and BTK by Compound (I).

[0226] The inhibition of HCK and BTK by compound (I) was evaluated by changes in the phosphorylation level of HCK at known functional sites in MYD88-mutated WM and ABC DLBCL cells, i.e., the phosphorylation of Thr on HCK was evaluated by PhosFlow. 209 ; BTK assessed by Western blot Tyr223 Ibrutinib was compared to HCK and BTK, and the results showed that ibrutinib moderately inhibited HCK phosphorylation, while compound (I) showed no significant effect on Thr phosphorylation. 209 Strong inhibition of HCK phosphorylation at Figure 3A ). It was observed that ibrutinib and compound (I) had a similar 223 The inhibition level of BTK was similar to that of Figure 3B The effects of compound (I) and ibrutinib on HCK and BTK phosphorylation were also determined by PhosFlow analysis in WM patient bone marrow tumor cells with MYD88 mutations in the presence of bone marrow microenvironment monocytes (N=4). Figure 3CWhole bone marrow mononuclear cells were treated with the indicated concentrations of ibrutinib, A419259, or compound (I) for 2.0 h and analyzed using BD PhosFlow TM Fixation buffer I (BD Biosciences) at 37°C for 10 min and BD Perm / Wash TM The cells were permeabilized with buffer I (BD Biosciences) and then stained with mouse anti-human CD20 (APC-cy7) specific antibody (BD Biosciences) together with rabbit anti-human p-HCK specific antibody, followed by donkey anti-rabbit IgG (Alexa Fluor). 647) secondary antibody or PE-labeled p-BTK antibody staining. + p-HCK or p-BTK levels were analyzed on LPC populations.

[0227] Example 3. Cellular efficacy of compound (I) in MYD88 wild-type (MYD88WT) and MYD88 mutant (MYD88L265P) B-cell lymphoma cells.

[0228] The antitumor effect of compound (I) was also evaluated in MYD88-mutated WM and ABC DLBCL cell lines as well as MYD88 wild-type GCB DLBCL, Burkitt's lymphoma, and multiple myeloma cell lines. Compound (I) showed targeted antitumor activity against MYD88 mutations relative to wild-type cell lines ( Figure 4A The antitumor activity of compound (I) was also evaluated using primary MYD88-mutated bone marrow-derived CD19-positive lymphoplasmacytic lymphoma cells (LPCs) from WM patients and compared against healthy donor CD19+ B cells. The activity of compound (I) was compared with ibrutinib in these experiments. Although ibrutinib showed a modest apoptotic effect on primary LPCs, compound (I) produced potent apoptosis of MYD88-mutated WM patient LPCs ( Figure 4B Compound (I) showed no apoptotic effect on healthy donor CD19+B cells ( Figure 4B ). PhosFlow analysis showed that the anti-apoptotic effect of compound (I) corresponds to the expression of p-HCK (T209) and p-BTK in LPC of WM patients. Y223 The reduction ( Figure 3C ).

[0229] In MYD88 mutant or wild-type B-cell lymphoma or multiple myeloma cell lines, the expression of MYD88 mutant or wild-type MYD88 mutant B-cell lymphoma or multiple myeloma cell lines was detected by CellTiter-Glo® after 72 h of treatment with serially diluted compound (I) or A419259. TMCell viability assays were used to measure in vitro cellular efficacy (drug dose-response). The blue line at 50 on the Y axis represents the EC50 level for each cell line at the corresponding drug concentration on the X axis ( Figure 4A ).

[0230] Example 4. Effect of Compound (I) on HCK T333M The resistance of the gatekeeper mutants suggests that HCK is a key target of compound (I) in MYD88 mutant WM cells.

[0231] Generation of HCK gatekeeper mutant HCK T333M , and rescue experiments were performed in BCWM.1 cells with MYD88 mutation. Compared with BCWM.1 cells transduced with vector or wild-type HCK, HCK T333M Expression of induced a more than 20-fold increase in resistance to compound (I) Figure 5A ). In addition, in the presence of compound (I), HCK T333M Not HCK WT The expression of HCK leads to the sustained activation of Figure 5B ). BCWM.1 cells expressing HCKWT or HCKT333M were also evaluated with compound (I) and found to contain BTK, AKT, ERK1 / 2 and SYK in the activated state. WT The expression of pBTK, pAKT, pERK1 / 2 and p-SYK was blocked in a dose-dependent manner in the expressing cells ( Figure 5C In contrast, the attenuation of pBTK, pAKT, pERK1 / 2, and pSYK was observed in HCK-expressing cells. T333M pBTK continued to show compound (I) in the presence of HCK T333M 1 cells, consistent with its role as a direct inhibitor of BTK activity ( Figure 5C ). Taken together, the above data indicate that HCK and BTK are on target and have biologically important sequelae in response to Compound (I) activity, including inhibition of multiple downstream pro-survival signaling pathways associated with HCK and BTK.

[0232] Figure 6 shows that HCK gatekeeper mutant (HCK T333M) Rescue compound (I)-induced cell death and block HCK activation and its downstream signaling in MWCL-1WM cells. After 72 hours of compound (I) treatment, vehicle or HCK alone WT or HCK T333M Dose response of transduced MWCL-1 cells ( Fig. 6A ). In the case of carrier only, HCK WT or HCK T333MRelative pHCKY411 levels ( Figure 6B After treatment with the indicated concentration of compound (I) for 1.0 hour, WT or HCK T333M Analysis of pBTK by Western Blot in Transduced BCWM.1 Cells Y223 , pSYK Y525 / 526 , p-AKT S473 、p-ERK1 / 2 T202 / Y204 Express( Figure 6C ). The expression levels of total BTK, SYK, AKT and ERK1 / 2 and the protein loading control GAPDH in these cells are also shown.

[0233] FIG. 7 shows that the HCK gatekeeper mutant (HCK T333M) Rescue compound (I)-induced cell death and block HCK activation and its downstream signaling in TMD8ABC DLBCL cells. After 72 hours of compound (I) treatment, vehicle or HCK alone WT or HCK T333M Dose response of transduced TMD7 cells ( Fig. 7A ). In the case of carrier only, HCK WT or HCK T333M Relative pHCK resolved by PhosFlow analysis in MYD88 mutant-transduced BCWM.1 cells treated with indicated concentrations of compound (I) for 1.0 h Y411 level( Figure 7B After treatment with the indicated concentration of compound (I) for 1.0 hour, WT or HCK T333M Analysis of pBTK by Western Blot in Transduced BCWM.1 Cells Y223 , pSYK Y525 / 526 , p-AKT S473 、p-ERK1 / 2 T202 / Y204 Express( Figure 7C ). The expression levels of total BTK, SYK, AKT and ERK1 / 2 and the protein loading control GAPDH in these cells are also shown.

[0234] Example 5. In vitro and in vivo overview of the pharmacokinetic properties of Compound (I).

[0235] Figure 8 Further description of the in vitro and in vivo pharmacokinetic properties of Compound (I) is shown. Incubation in liver microsomes showed excellent stability (T1 / 2 Incubation in liver microsomes showed excellent stability in a variety of species including humans, mice and rats (T 1 / 2 About or longer than 60 minutes). The mouse pharmacokinetic parameters of compound (I) after intravenous administration of 2 mg / kg, 10 mg / kg and 25 mg / kg are shown. Consistent with the long microsomal stability of compound (I), mouse pharmacokinetic studies showed that the bioavailability after oral administration of 10 mg / kg and 25 mg / kg was 49% to 55%, respectively. The serum half-life after intravenous administration was 11.5 hours, while after oral administration of 10 mg / kg and 25 mg / kg, it was 15.1 and 16.9 hours, respectively. In vitro metabolism studies of CYP inhibition (% at 10 μM) were as follows: -27.9 (1A), 64.5 (2B6), -3.8 (2C8), -10.9 (2C9), 0.8 (2C19), 24.9 (2D6), -22.7 / -4.9 (3A). Ames genetic toxicity assays were performed on four test strains of Salmonella typhimurium (TA98, TA100, TA1535, and TA1537) in the presence and absence of metabolic activation in rat liver S9 fractions, and the results were all negative in the 50 μM range. The hERG test (patch clamp) for cardiotoxicity was negative at 16 μM. Tolerance studies showed that compound (I) was well tolerated at daily oral doses of up to 75 mg / kg in NOD-SCID mice, and no adverse events were observed for more than 6 weeks of dosing. Plasma protein binding showed comparable levels in different species, as follows: mouse (97.7%); rat (95.5%); dog (95.0%); and human (95.7%).

[0236] Consistent with the low metabolic rate observed in the microsomal assay, compound (I) provided 17.4 mL min in mice. -1 kg -1 When administered orally at 10 mg / kg and 25 mg / kg, compound (I) was well absorbed with excellent exposure (AUC = 5.2, 11.4 μM h, respectively), high bioavailability (%F = 55% and 49%, respectively) and desirable drug clearance (T 1 / 2 = 15.1 to 16.9 hours).

[0237] When incubated with purified microsomes, compound (I) exhibited t 1 / 2 =106.4 minutes (mouse), t 1 / 2 =>120 minutes (rat), t 1 / 2 = 60 minutes (dog) and t 1 / 2= 49.5 minutes (human) half-life. In contrast, A419259 has a very long T when administered orally. 1 / 2 (176.0 hours)( Fig. 9 ), which could pose a problem for clinical development.

[0238] Example 6. Mean tumor volume curves and survival curves of NOD-SCID mice implanted with TMD8 ABCDLBCL tumor cells expressing wild-type BTK treated with ibrutinib, A419259 or Compound (I).

[0239] Figure 11 shows the mean tumor volume curve ( Fig.11A ), tumor volume statistics on day 33 ( Fig. 11B ) and the survival curve ( Fig. 11C ). TMD8 tumor cells were implanted subcutaneously into NOD-SCID mice, with 8 mice per group. After tumors were established and reached approximately 200 cubic millimeters, mice were treated orally with vehicle control or ibrutinib or A419259 or compound (I) at the indicated concentrations on a once-daily schedule. Tumors were measured twice a week with an electronic caliper ( Fig.11A ). Tumor volumes in all treatment groups were significantly smaller than those in mice receiving vehicle control on day 33 (P<0.005 by "Wilcoxon Rank Sum Test", P<0.0001 by "Tukey's Multiple Comparison Test") Fig. 11B ). Treatment was stopped after completing week 6 of treatment, and tumor volume assessment continued for all surviving mice until week 16. Mouse survival was also followed by Kaplan-Meier survival curves until the end of the study at week 16.

[0240] Example 7. Compound (I) overcomes BTK in vitro in BCWM.1WM and TMD8 ABC DLBCL cell models C481S Mutations causing ibrutinib resistance.

[0241] Figure 12 shows that only in vector, expression of BTK WT or BTK C481S TMD8 ABC-DLBCL or BCWM.1WM cells transduced with lentiviral vectors were expressed by CellTiter-Glo TM Cell viability assay measures drug dose-response. C481S Unlike ibrutinib, which is resistant to mutants, compound (I) is only expressed in vector, BTKWT or BTK C481S The results showed that the BCWM.1 and TMD8 cells produced similar cellular efficacy. Fig. 12AWe measured apoptosis analysis by flow cytometry with Annexin-V / PI staining, and the percentage of apoptotic cells was marked. WT or BTK C481S Similar cell efficacy was observed in BCWM.1 and TMD8 cells ( Fig. 12B ).

[0242] Example 8. Compound (I) blocks ibrutinib-resistant BTK C481S HCK, BTK, and downstream ERK1 / 2 activation in mutant BCWM.1WM and TMD8 ABC DLBCL cell lines.

[0243] Fig.13A Compound (I) shows the effects of BCWM.1 or TMD8 cells, TMD8 cells transduced with lentiviral vector alone, or cells expressing BTK WT or BTK C481S PhosFlow analysis of the effects of mutants on HCK phosphorylation levels in cells. Cells were treated with the indicated concentrations of ibrutinib, A419259, or compound (I) for 1.0 h and analyzed using BD PhosFlow TM Fixation buffer I (BD Biosciences) was used for 10 min at 37°C and the cells were fixed with BD Perm / Wash TM Buffer I (BD Biosciences) was used for permeabilization, followed by staining with rabbit anti-human p-HCK specific antibody (Abcam), followed by donkey anti-rabbit IgG (Alexa Fluor Compound (I) and A419259 effectively blocked HCK phosphorylation, while ibrutinib moderately reduced the phosphorylation of vehicle and BTK WT or BTK C481S HCK phosphorylation in transduced BCWM.1 or TMD8 cells.

[0244] Fig. 13B The effect of compound (I) on the expression of BTK using only lentiviral vector WT or BTK C481S Western blot of the effects of mutant lentiviral vectors on BTK phosphorylation levels in BCWM.1 or TMD8 cells. Cells were treated with the indicated concentrations of ibrutinib or compound (I) for 1.0 h and lysed with protein lysis buffer. Both compound (I) and ibrutinib effectively blocked the expression of both vector and BTK. WT BTK and ERK 1 / 2 phosphorylation in transduced BCWM.1 or TMD8 cells. However, only compound (I) effectively blocked BTK C481SBTK and ERK 1 / 2 phosphorylation in transduced BCWM.1 or TMD8 cells was not blocked by ibrutinib.

[0245] Example 9. Pharmacodynamic studies show that HCK and BTK are ibrutinib-resistant in subcutaneous xenografts C481S Activity of TMD8ABC-DLBCL cells in NOD-SCID mice.

[0246] Ibrutinib-resistant TMD8-BTK C481S Tumor cells were implanted subcutaneously in NOD-SCID mice, with 8 mice per group. After tumors were established and reached approximately 300 cubic millimeters, mice were orally treated with vehicle control, ibrutinib, A419259, or the indicated concentrations of compound (I) on a once-daily schedule. Tumors were measured twice a week with an electronic caliper ( FIG. 14 ). GFP was excised 6 and 24 hours after oral administration of compound (I) at the indicated doses. + pHCKY411( Fig.14A ) and pBTKY223( Fig. 14B ) (n=3 in each group). *p<0.05; **p<0.01; ***p<0.005; ****p<0.0001.

[0247] Example 10. Effect of Compound (I) on Ibrutinib-resistant TMD8-BTK C481S Mean tumor volume curves in the two transplantation models.

[0248] Ibrutinib-resistant TMD8-BTK C481S Tumor cells were implanted subcutaneously in NOD-SCID mice, with 8 mice per group. After tumors were established and reached approximately 200 cubic millimeters, mice were treated orally with vehicle control or ibrutinib (50 mg / kg) or compound (I) (50 mg / kg or 75 mg / kg) on ​​a daily schedule. After daily oral administration of vehicle control, ibrutinib (50 mg / kg) or compound (I) (50 or 75 mg / kg), mice carrying ibrutinib-resistant BTK C481S Efficacy study in NOD-SCID mice (n=8 per group) with TMD8 cells. Tumor volume (mm 3 ) and reported as mean volume ± SEM ( Fig.15A ). Comparison of tumor volume on day 29. The p-values ​​for group comparisons are shown ( Fig. 15B The survival curve was estimated using the Kaplan-Meier method. The median survival time (days) of the groups was displayed using Prism software. The log-rank comparison between the groups was P = 0.0007 ( Fig. 15C ).

[0249] After daily oral administration of vehicle control, ibrutinib (50 mg / kg), A419259 (50 mg / kg), or compound (I) (50 mg / kg), mice carrying cells expressing ibrutinib-resistant BTK C481S Efficacy study in NOD-SCID mice (n=8 per group) with TMD8 cells. Tumor volume (mm 3 ) and reported as mean volume ± SEM ( Fig.16A ). Comparison of tumor volume on day 33. The p-values ​​for group comparisons are shown ( Fig. 16B The survival curve was estimated using the Kaplan-Meier method. The median survival time (days) of the groups was displayed using Prism software. The log-rank comparison between the groups was P<0.0001 ( Fig. 16C ).

[0250] Example 11. Combination index (CI) of Compound (I) and venetoclax (BCL-2 inhibitor) in MYD88 mutated Waldenstrom's macroglobulinemia (BCWM.1) cells.

[0251] In vitro studies evaluated compound (I) and venetoclax in native (untransduced), BTKWT- and BTK-expressing C481S Figure 3 Synergistic interactions of MYD88-mutated BCWM.1WM and TMD8 ABC DLBCL cells. Combination index (CI) and normalized isobologram analysis are depicted. CI < 1 (indicated by red shading) or points under the slash in the isobologram indicate a synergistic effect of the combination ( Fig.17 ).

[0252] Example 12. Compound (I) and venetoclax (BCL-2 inhibitor) in non-transduced native MYD88-mutated Waldenstrom's macroglobulinemia (BCWM.1) and ABC DLBCL (TMD8) cells and in BTK WT or BTK C481S Combination index (CI) in transduced BCWM.1 or TMD8 cells.

[0253] Figure 18 shows BTK in ibrutinib resistance C481S Effect of the combination of Compound (I) and Venetoclax on tumor volume and survival in a TMD8 ABC DLBCL xenograft mouse model. Following daily oral administration of vehicle control, venetoclax (50 mg / kg), Compound (I) (30 mg / kg), or the combination of venetoclax (50 mg / kg) and Compound (I) (30 mg / kg), tumor volume and survival were observed in mice carrying a TMD8 ABC DLBCL xenograft expressing ibrutinib-resistant BTK. C481S Efficacy study in NOD-SCID mice (n=8 per group) with TMD8 cells. Tumor volume (mm 3) and reported as mean volume ± SEM ( Fig.18A ). Comparison of tumor volume on day 22. The p-values ​​for group comparisons are shown ( Fig.18B The survival curve was estimated using the Kaplan-Meier method. The median survival time (days) of the groups was displayed using Prism software. The log-rank comparison between the groups was P = 0.0020 ( Fig.18C ).

[0254] As described above, the combination of Compound (I) and venetoclax produces synthetic lethality in MYD88-mutated lymphoma cells.

[0255] Example 13. HCK activation status in chronic lymphocytic leukemia patients.

[0256] Fig.19 The HCK activation status of CLL patients was evaluated in primary tumor cells from peripheral blood mononuclear cells (PBMCs) of previously treated or untreated patients. PBMCs were separated by Ficoll-Paque and analyzed by BD PhosFlow TM Fixation buffer I (BD Biosciences) was used for 10 min at 37°C and the cells were fixed with BD Perm / Wash TM The cells were permeabilized with buffer I (BD Biosciences) and then stained with mouse anti-human CD20 (APC-cy7) and CD5 (BV421) specific antibodies (BD Biosciences) together with rabbit anti-human p-HCK specific antibodies, followed by donkey anti-rabbit IgG (Alexa Fluor). 647) secondary antibody or PE-labeled p-BTK antibody staining. + CD5 + The p-HCK or p-BTK levels were analyzed in groups.

[0257] Example 14. In vitro studies using ibrutinib or Compound (I) on peripheral blood mononuclear cells isolated from ibrutinib-relapsed CLL patients.

[0258] Fig. 20 Isolation of PBMCs from an ibrutinib-relapsed CLL patient by Ficoll-Pac and separation of CD19 by magnetic beads are shown. + cells and BTK by Sanger sequencing C481S The genotyping results later showed that the patient had approximately 70% CD19 + Cells carrying BTK C481S Mutation (T>A).

[0259] Freshly isolated PBMCs were treated with ibrutinib or compound (I) overnight in cell culture medium supplemented with 10% FBS. After drug treatment, PBMCs were stained with Annexin V-FITC and PI and mouse anti-human CD19 (APC-cy7) and CD5 (BV421) specific antibodies (BD Biosciences). CD19 + CD5 + Apoptotic cells were measured in populations.

[0260] Example 15. In BTK WT or BTK C481S Western blot analysis of BTK, LYN, ERK1, and ERK2 in BCWM.1 cells after treatment with ibrutinib, dasatinib, A419259, or Compound (I).

[0261] Fig.21 Shown in BTK WT or BTK C481S Western blot analysis of phosphorylation levels of BTK, LYN, and ERK1 / 2 in transduced BCWM.1 cells treated with the indicated concentrations of ibrutinib, dasatinib, A419259, and compound (I) for 1 hour. Total protein of each kinase and GAPDH was used for protein expression and loading control. Compound (I) effectively blocked the expression of BTK WT and BTK C481S BTK, LYN, and ERK1 / ERK2 phosphorylation in mutant BCWM.1 cells.

[0262] Example 16. Synthesis of compound (I).

[0263] Compound (I) was synthesized according to the following scheme.

[0264]

[0265] (1s,4s)-4-(4-Benzylpiperazin-1-yl)cyclohexanol (SM1)

[0266] cis-4-aminocyclohexanol hydrochloride (2.0 g, 17.3 mmol), N-benzyl-2-chloro-N-(2-chloroethyl)ethanamine (5.5 g, 20.76 mmol) and NaHCO 3 A mixture of (5.7 g, 69.2 mmol) in ethanol (30 mL) was stirred at 90 ° C for 5 hours. The mixture was then concentrated in vacuo, and the residue was diluted with water (100 mL) and extracted with ethyl acetate (150 mL×2). The combined organic phase was then washed with brine (50 mL×2) and dried over anhydrous Na 2 SO 4Dry, concentrate, and purify by silica gel column chromatography (DCM / methanol=10 / 1) to give SM1 as a yellow oil (2.8 g, yield 57%). LCMS (m / z): 275.2 [M+H] + .

[0267] (1s,4s)-4-(piperazin-1-yl)cyclohexanol (SM2)

[0268] SM1 (2.7 g, 9.85 mmol), CH 3 A mixture of COOH (0.5 mL) and Pd / C (10%, 270 mg) in i-PrOH (20 mL) was heated at room temperature under H 2 The mixture was stirred at 40 °C (1 atm) for 24 h. Afterwards, the mixture was filtered and the filtrate was concentrated to give crude SM2 as a yellow oil (1.5 g, yield 83%). LCMS (m / z): 185.1 [M+H] + .

[0269] 4-((1s,4s)-4-Hydroxycyclohexyl)piperazine-1-carboxylic acid tert-butyl ester (SM3)

[0270] SM2 (1.5 g, 8.1 mmol), (Boc 2 A mixture of O (4.0 mL, 16.2 mmol) and DIPEA (4.0 mL) in THF (20 mL) was stirred at room temperature for 2 hours. After that, the mixture was concentrated to give crude SM3 as a brown oil (2.5 g, yield 80%). LCMS (m / z): 285.2 [M+H] + .

[0271] tert-Butyl 4-((1s,4s)-4-(methylsulfonyloxy)cyclohexyl)piperazine-1-carboxylate (SM4)

[0272] MsCl (1.0 mL) was added dropwise to a mixture of SM3 (2.5 g, 8.1 mmol) and DIPEA (4.0 mL) in DCM (20 mL) at 0 °C. The mixture was then stirred at room temperature for 4 hours and concentrated to remove the solvent, and the residue was diluted with water (100 mL) and extracted with ethyl acetate (150 mL×2). The combined organic extracts were then washed with brine (50 mL×2) and washed with anhydrous Na 2 SO 4 Drying, filtration and concentration gave crude SM4 as a brown oil (600 mg, 20% yield). LCMS (m / z): 363.0 [M+H] + .

[0273] 4-((1r,4r)-4-(4-amino-3-bromo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexyl)piperazine-1-carboxylic acid tert-butyl ester (SM5)

[0274] 3-Bromo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (353 mg, 1.65 mmol), SM4 (600 mg, 1.65 mmol) and Cs 2 CO 3 A mixture of (1.0 g, 3.3 mmol) in DMF (15 mL) was stirred at 100 °C for 16 hours. Afterwards, the reaction mixture was cooled to room temperature, diluted with brine (100 mL), and extracted with ethyl acetate (50 mL x 2). The combined organic extracts were purified by anhydrous Na 2 SO 4 Dry, filter, concentrate and purify by silica gel column chromatography (DCM / methanol=10 / 1) to give SM5 as a yellow solid (300 mg, yield 38%). LCMS (m / z): 480.0 [M+H] + .

[0275] 4-((1r,4r)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexyl)piperazine-1-carboxylic acid tert-butyl ester (SM6)

[0276] SM5 (60 mg, 0.12 mmol), 4,4,5,5-tetramethyl-2-(4-phenoxyphenyl)-1,3,2-dioxaborolane (56 mg, 0.18 mmol), Pd(PPh 3 ) 2 Cl 2 (9.0 mg, 0.012 mmol) and K 3 PO 4 (51 mg, 0.24 mmol) in THF (2.0 mL) and H 2 O (0.5 mL) was stirred at 60 ° C for 16 hours. After that, the mixture was filtered, and the filtrate was diluted with brine (50 mL) and extracted with ethyl acetate (50 mL×2). The combined organic extracts were then washed with anhydrous Na 2 SO 4 Dry, filter, concentrate and purify by silica gel column chromatography (DCM / methanol=10 / 1) to give SM6 as a brown oil (40 mg, yield 58%). LCMS (m / z): 570.4 [M+H] + .

[0277] 3-(4-Phenoxyphenyl)-1-((1r,4r)-4-(piperazin-1-yl)cyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (SM7)

[0278] To a solution of SM6 (40 mg, 0.07 mmol) in DCM (5.0 mL) was added a solution of HCl in 1,4-dioxane (4 M, 2.0 mL), and the mixture was stirred at room temperature for 2 hours. Thereafter, the reaction mixture was concentrated in vacuo, and the residue was diluted with DCM (30 mL). The residue was washed with saturated Na 2 CO 3 The solution was adjusted to pH 8 and filtered, the filtrate was concentrated and analyzed by preparative HPLC (C18 column, CH 3 CN / H 2 O, containing 0.05% TFA) to give SM7 as a white solid (15.2 mg, yield 47.5%). LCMS (m / z): 470.1 [M+H]+; 1H NMR (400 MHz, DMSO-d6): δ 8.23 ​​(s, 1H), 7.64 (d, J = 8.8 Hz, 2H), 7.45-7.41 (m, 2H), 7.20-7.11 (m, 5H), 4.64-4.63 (m, 1H), 2.85-2.81 (m, 4H), 2.57 (s, 2H), 2.49-2.39 (m, 2H), 2.04-1.90 (m, 6H), 1.50-1.47 (m, 2H).

[0279] 1-((1r,4r)-4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (I)

[0280] SM7 (400 mg, 0.85 mmol), (CH 2 O) n (76 mg, 0.85 mmol) and CH 3 A mixture of COOH (0.5 mL) in methanol (10.0 mL) was stirred at room temperature for 3 h, and then NaBH 4 (97 mg, 2.55 mmol), the mixture was stirred at room temperature for 16 hours and concentrated in vacuo, the residue was diluted with brine (100 mL) and extracted with ethyl acetate (150 mL×2), the combined organics were washed with anhydrous Na 2 SO 4 Dry, filter, concentrate, and purify by preparative HPLC to give compound (I) as a white solid (150 mg, yield 36%). LCMS (m / z): 484.1 [M+H] + ;1 H-NMR (400MHz, DMSO-d6): δ8.23(s,1H),7.64(d,J=8.8Hz,2H),7.43(t,J=8.0Hz,2H),7.20-7.11(m, 5H),4.63(m,1H),2.52(m,3H),2.37-2.30(m,6H),2.13(s,3H),2.03-1.91(m,6H),1.47-1.43(m,2H).

[0281] Equivalents and scope

[0282] In the claims, articles such as "a," "an," and "the" may refer to one or more than one, unless indicated to the contrary or obvious from the context. Claims or descriptions containing "or" between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process, unless indicated to the contrary or otherwise obvious from the context. The present disclosure includes embodiments in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process. The present disclosure includes embodiments in which more than one or all of the group members are present in, employed in, or otherwise relevant to a given product or process.

[0283] In addition, the present disclosure includes all changes, combinations and arrangements in which one or more restrictions, elements, clauses and descriptive terms from one or more listed claims are introduced into another claim. For example, any claim subordinate to another claim can be modified to be included in one or more restrictions found in any other claim subordinate to the same basic claim. In the case where the element is presented as a list, for example, in a Markush group format, each subgroup of the element is also disclosed, and any element can be removed from the group. It should be understood that, in general, when the disclosure or aspects described herein are referred to as including specific elements and / or features, some embodiments described herein or aspects described herein are composed of such elements and / or features or are substantially composed of such elements and / or features. For simplicity, these embodiments are not specifically described in the same words herein. It should also be noted that the terms "including" and "containing" are intended to be open and allow the inclusion of additional elements or steps. When a range is given, end values ​​are included. In addition, unless otherwise indicated or otherwise obvious from the context and understanding of one of ordinary skill in the art, values ​​expressed as ranges may assume any specific value or sub-range within the stated range in the various embodiments described herein, to the tenth of the unit of the lower limit of the range, unless the context clearly indicates otherwise.

[0284] This application relates to various published patents, published patent applications, journal articles and other publications, all of which are incorporated herein by reference. If there is a conflict between any incorporated reference and this specification, the present specification shall prevail. In addition, any specific embodiment of the present disclosure that falls within the prior art may be explicitly excluded from any one or more claims. Because such embodiments are considered to be known to those of ordinary skill in the art, they may be excluded, even if the exclusion is not explicitly set forth herein. Regardless of whether the prior art is involved, any specific embodiment described herein may be excluded from any claim for any reason.

[0285] Those skilled in the art will recognize or be able to ascertain many equivalents to the specific embodiments described herein using only routine experimentation. The scope of the current embodiments described herein is not intended to be limited to the above description, but rather to that set forth in the appended claims. Those of ordinary skill in the art will appreciate that various changes and modifications may be made to this specification without departing from the spirit or scope of the present disclosure as defined by the following claims.

Claims

1. A compound (I) of the following formula: Or a pharmaceutically acceptable salt thereof for preparing a medicament for treating a proliferative disease associated with a mutation in the MYD88 protein, wherein the proliferative disease is Waldenstrom's macroglobulinemia WM or activated B-cell-like diffuse large B-cell lymphoma ABC-DLBCL.

2. A compound (I) of the following formula: or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating a proliferative disease in a subject having a mutation in the MYD88 protein, wherein the proliferative disease is Waldenstrom's macroglobulinemia WM or activated B-cell-like diffuse large B-cell lymphoma ABC-DLBCL.

3. The use according to claim 1 or 2, wherein the disease is associated with abnormal activity of the hematopoietic cell kinase HCK.

4. The use according to claim 1 or 2, wherein the disease is associated with abnormal activity of LYN proto-oncogene tyrosine kinase.

5. The use according to claim 1 or 2, wherein the disease is associated with abnormal activity of Bruton's tyrosine kinase BTK.

6. The use according to claim 1 or 2, wherein the disease is associated with a mutation in the BTK protein.

7. The use according to claim 6, wherein the mutant BTK protein is C481S mutant BTK.

8. The use according to claim 3, wherein the HCK is a mutated HCK.

9. The use according to claim 6, wherein the BTK is mutated at Cys481.

10. The use according to claim 5, wherein the BTK is resistant to inhibition by ibrutinib or a pharmaceutically acceptable salt thereof.

11. The use according to claim 6, wherein the BTK is resistant to inhibition by ibrutinib or a pharmaceutically acceptable salt thereof.

12. The use according to claim 1 or 2, wherein the subject is resistant to treatment with a BTK inhibitor.

13. The use according to claim 12, wherein the BTK inhibitor is ibrutinib, CC-292, ONO-4059, evobutinib, septinib, BGB-3111, HM71224 or ACP-196, or a pharmaceutically acceptable salt thereof.

14. The use according to claim 12, wherein the BTK inhibitor is ibrutinib, or a pharmaceutically acceptable salt thereof.

15. The use according to claim 1 or 2, wherein the subject is a mammal.

16. The use according to claim 1 or 2, wherein the subject is a human.

17. The use according to claim 1 or 2, wherein the subject has previously been treated with a BTK inhibitor.

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