HCK AS A THERAPEUTIC TARGET IN MYD88 MUTATED DISEASES

Abstract

The present disclosure provides compounds of Formula (I), and pharmaceutically acceptable salts, solvates, hydrates, polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeled derivatives, and prodrugs thereof. The provided compounds may be kinase (e.g., HCK, BTK, LYN, BLK, FRK) inhibitors. Also provided are pharmaceutical compositions and kits including the provided compounds. Further provided are methods of using the provided compounds, pharmaceutical compositions, and kits (e.g., for treating diseases (e.g., proliferative diseases) in a subject in need thereof).

##STR00001##

Claims

1. A compound of Formula (I): ##STR00101## or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof, wherein: R is optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, —O-optionally substituted carbocyclyl, —CH.sub.2—O-optionally substituted carbocyclyl, —O-optionally substituted aryl, —CH.sub.2—O-optionally substituted aryl, —O-optionally substituted heteroaryl, or —CH.sub.2—O-optionally substituted heteroaryl; R.sup.1 is hydrogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or a nitrogen protecting group; each occurrence of R.sup.2 is independently hydrogen, halogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, —OR.sup.a, —N(R.sup.a).sub.2, or —SR.sup.a, wherein R.sup.a is independently selected from hydrogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, an oxygen protecting group when attached to an oxygen atom, and a sulfur protecting group when attached to a sulfur atom; each occurrence of R.sup.3 is independently hydrogen, halogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, —OR.sup.b, —N(R.sup.b).sub.2, or —SR.sup.b, wherein R.sup.b is independently selected from hydrogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, an oxygen protecting group when attached to an oxygen atom, and a sulfur protecting group when attached to a sulfur atom; J is a bond or alkynylene; m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, as valency permits; n is 1, 2, 3, 4, 5, 6, 7, or 8, as valency permits; A is an optionally substituted mono- or bicyclic-heteroaryl, or optionally substituted bicyclic heterocyclyl; and provided that when J is a bond, R is substituted phenyl, R.sup.1 is methyl, R.sup.2 is H, and R.sup.3 is H, then ring A is not of the formula: ##STR00102##

2. The compound of claim 1, wherein ring A is of the formula: ##STR00103## or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof, wherein: R.sup.4 is independently selected from hydrogen, halogen, optionally substituted acyl, optionally substituted alkyl, —CN, —SCN, —NO.sub.2, —C(═O)R.sup.b, —C(═O)N(R.sup.b).sub.2, —OR.sup.b, —N(R.sup.b).sub.2, —C(═NR.sup.b)R.sup.b, —C(═NR.sup.b)OR.sup.b, —C(═NR.sup.b)N(R.sup.b).sub.2, —C(═O)OR.sup.b, —NR.sup.bC(═O)R.sup.b, —NR.sup.bC(═O)OR.sup.b, —NR.sup.bC(═O)N(R.sup.b).sub.2, —OC(═O)R.sup.b, —OC(═O)OR.sup.b, —OC(═O)N(R.sup.b).sub.2, or —SR.sup.b, wherein R.sup.b is independently selected from hydrogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, an oxygen protecting group when attached to an oxygen atom, and a sulfur protecting group when attached to a sulfur atom: R.sup.5 is independently selected from hydrogen, halogen, optionally substituted acyl, optionally substituted alkyl, —CN, —SCN, —NO.sub.2, —C(═O)R.sup.b, —C(═O)N(R.sup.b).sub.2, —OR.sup.b, —N(R.sup.b).sub.2, —C(═NR.sup.b)R.sup.b, —C(═NR.sup.b)OR.sup.b, —C(═NR.sup.b)N(R.sup.b).sub.2, —C(═O)OR.sup.b, —NR.sup.bC(═O)R.sup.b, —NR.sup.bC(═O)OR.sup.b, —NR.sup.bC(═O)N(R.sup.b).sub.2, —OC(═O)R.sup.b, —OC(═O)OR.sup.b, —OC(═O)N(R.sup.b).sub.2, or —SR.sup.b, wherein R.sup.b is independently selected from hydrogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, an oxygen protecting group when attached to an oxygen atom, and a sulfur protecting group when attached to a sulfur atom; R.sup.6 is independently selected from hydrogen, halogen, optionally substituted acyl, optionally substituted alkyl, —CN, —SCN, —NO.sub.2, —C(═O)R.sup.b, —C(═O)N(R.sup.b).sub.2, —OR.sup.b, —N(R.sup.b).sub.2, —C(═NR.sup.b)R.sup.b, —C(═NR.sup.b)OR.sup.b, —C(═NR.sup.b)N(R.sup.b).sub.2, —C(═O)OR.sup.b, —NR.sup.bC(═O)R.sup.b, —NR.sup.bC(═O)OR.sup.b, —NR.sup.bC(═O)N(R.sup.b).sub.2, —OC(═O)R.sup.b, —OC(═O)OR.sup.b, —OC(═O)N(R.sup.b).sub.2, or —SR.sup.b, wherein R.sup.b is independently selected from hydrogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, an oxygen protecting group when attached to an oxygen atom, and a sulfur protecting group when attached to a sulfur atom; X.sup.1 and X.sup.2 are independently ═N—, or ═C(R.sup.c)—, as valency permits, wherein R.sup.c is hydrogen, halogen, or substituted or unsubstituted C.sub.1-6 alkyl; X.sup.3 is N, C, or C(R.sup.d), as valency permits, wherein R.sup.d is hydrogen, halogen, or substituted or unsubstituted C.sub.1-6 alkyl; and p is 0 or 1, as valency permits.

3. The compound of claim 1, wherein the compound is selected from the group consisting of Formula (II): ##STR00104##

4-5. (canceled)

6. The compound of claim 3, wherein (a) X.sup.1 is ═N—; or (b) X.sup.1 is ═C(H)—; and (c) X.sup.2 is ═N—; or (d) X.sup.2 is ═CH—; and (e) X.sup.3 is N; or (f) X.sup.3 is C; or (g) X.sup.3 is CH; and (h) X.sup.4 is ═N—; or (i) X.sup.4 is ═CH—.

7-8. (canceled)

9. The compound of a claim 3, wherein the compound is of the formula: ##STR00105##

10-11. (canceled)

12. The compound of claim 3, wherein (a) is 0; or (b) p is 1; and (c) J is a bond; or (d) J is alkynylene; and (e) R is optionally substituted phenyl; or (f) R is 4-phenoxyphenyl; or (g) R is —CH.sub.2—O-substituted phenyl; or (h) R is —CH.sub.2—O-unsubstituted phenyl; and (i) R.sup.1 is optionally substituted alkyl; or (j) R.sup.1 is unsubstituted methyl; and (k) R.sup.2 is H; and (1) R.sup.3 is H; and (m) R.sup.4 is H; or (n) R.sup.4 is —C(O)NH.sub.2; and (o) R.sup.5 is NH.sub.2.

13-48. (canceled)

49. A compound of claim 1, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof, selected from the formula: ##STR00106## ##STR00107## ##STR00108##

50. A pharmaceutical composition comprising a compound of claim 1 and one or more pharmaceutically acceptable excipient.

51. The pharmaceutical composition of claim 50 further comprising an additional pharmaceutical agent.

52-54. (canceled)

55. The pharmaceutical composition of claim 51, wherein the additional pharmaceutical agent is venetoclax.

56. (canceled)

57. A method of treating a disease associated with an MYD88 mutation in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of claim 1, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

58. The method of claim 57, wherein the disease is a proliferative disease.

59. The method of claim 58, wherein the proliferative disease is cancer.

60-64. (canceled)

65. The method of claim 59, wherein the cancer is Waldenstrom's macro-globulinemia.

66-67. (canceled)

68. The method of claim 6759, wherein, the cancer is activated B-cell-like (ABC)-DLBCL.

69-89. (canceled)

90. The method of claim 59, wherein the cancer is associated with aberrant activity of one or more of HCK, -BTK, and LYN in a subject.

91. The method of claim 90, wherein the aberrant activity is overexpression.

92-106. (canceled)

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0116] The accompanying drawings, which constitute a part of this specification, illustrate several embodiments of the disclosure and together with the description, serve to explain the principles of the disclosure.

[0117] FIG. 1 shows pro-survival signaling driven by mutated MYD88 and the possible kinases targeted by the compounds of Formula (I).

[0118] FIG. 2 shows a dose-response curves for Compound I-4, as measured by CellTiter-Glo™ cell viability assay in MYD88 mutated BCWM.1, MWCL-1 WM cells; TMD8, HBL-1 ABC DLBCL cells; MYD88 wild type OCI-Ly7, OCI-Ly19 GCB DLBCL cells; Ramos Burkitt's lymphoma cells; and multiple myeloma cells.

[0119] FIG. 3 shows a dose-response curves for Compound I-1, as measured by CellTiter-Glo™ cell viability assay in MYD88 mutated BCWM.1, MWCL-1 WM cells; TMD8, HBL-1 ABC DLBCL cells; MYD88 wild type OCI-Ly7, OCI-Ly19 GCB DLBCL cells; Ramos Burkitt's lymphoma cells; and multiple myeloma cells.

[0120] FIG. 4 shows the drug dose-response for ibrutinib and Compound I-4 in BTK WT or BTK C481S expressing lentiviral vector transduced TMD8 ABC-DLBCL or BCWM.1 WM cells.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION

[0121] Kinases are implicated in a wide variety of diseases, e.g., proliferative diseases. Provided herein are compounds of Formula (I), and pharmaceutically acceptable salts, solvates, hydrates, polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeled derivatives, and prodrugs thereof. The provided compounds may be kinase inhibitors. In certain embodiments, the kinase being targeted is an SRC family kinase (e.g., HCK, LYN, BLK, FRK). In certain embodiments, the kinase being targeted is a Tec family kinase (e.g., BTK). Also provided are pharmaceutical compositions and kits comprising the provided compounds. Further provided are methods of using the compounds, pharmaceutical compositions, and kits for treating a disease (e.g., proliferative disease) in a subject in need thereof. In certain embodiments, the disease is a proliferative disease (e.g., an IgM gammopathy (e.g., an 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, stomach cancer, lymphoma (e.g., B-cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgM secreting lymphoplasmacytic lymphoma (i.e., Waldenström'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 myelomas (e.g., IgM multiple myeloma)), and leukemia (e.g., chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia, myelogenous leukemia (e.g., chronic myelogenous leukemia, acute myelogenous leukemia (e.g., mast cell leukemia), myeloproliferative diseases (e.g., myelodysplastic syndrome))))) Further provided are methods of using the provided compounds, pharmaceutical compositions, and kits for inhibiting the activity of a kinase in a subject in need thereof, in a biological sample, or in a cell.

Compounds

[0122] In one aspect of the present disclosure, provided herein are compounds of Formula (I):

##STR00009##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof, wherein:

[0123] R is optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, —O-optionally substituted carbocyclyl, —CH.sub.2—O-optionally substituted carbocyclyl, —O-optionally substituted aryl, —CH.sub.2—O-optionally substituted aryl, —O-optionally substituted heteroaryl, or

—CH.sub.2—O-optionally substituted heteroaryl;

[0124] R.sup.1 is hydrogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or a nitrogen protecting group;

[0125] each occurrence of R.sup.2 is independently hydrogen, halogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, —OR.sup.a, —N(R.sup.a).sub.2, or —SR.sup.a, wherein R.sup.a is independently selected from hydrogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, an oxygen protecting group when attached to an oxygen atom, and a sulfur protecting group when attached to a sulfur atom;

[0126] each occurrence of R.sup.3 is independently hydrogen, halogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, —OR.sup.b, —N(R.sup.b).sub.2, or —SR.sup.b, wherein R.sup.b is independently selected from hydrogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, an oxygen protecting group when attached to an oxygen atom, and a sulfur protecting group when attached to a sulfur atom;

[0127] J is a bond or alkynylene;

[0128] m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, as valency permits;

[0129] n is 1, 2, 3, 4, 5, 6, 7, or 8, as valency permits;

[0130] A is an optionally substituted mono- or bicyclic-heteroaryl, or optionally substituted bicyclic heterocyclyl; and

[0131] provided that when J is a bond, R is substituted phenyl, R.sup.1 is methyl, R.sup.2 is H, and R.sup.3 is H, then ring A is not of the formula:

##STR00010##

[0132] Formula (I), as described herein, contains the substituent R. In certain embodiments, R is optionally substituted alkynyl. In certain embodiments, R is optionally substituted carbocyclyl. In certain embodiments, R is optionally substituted heterocyclyl. In certain embodiments, R is optionally substituted aryl. In certain embodiments, R is optionally substituted phenyl. In certain embodiments, R is 4-phenoxyphenyl. In certain embodiments, R is optionally substituted heteroaryl. In certain embodiments, R is —O-optionally substituted carbocyclyl. In certain embodiments, R is —CH.sub.2—O-optionally substituted carbocyclyl. In certain embodiments, R is —O-optionally substituted aryl. In certain embodiments, R is —CH.sub.2—O-optionally substituted aryl. In certain embodiments, R is —CH.sub.2—O-optionally substituted phenyl. In certain embodiments, R is —CH.sub.2—O-unsubstituted phenyl. In certain embodiments, R is —O-optionally substituted heteroaryl. In certain embodiments, R is —CH.sub.2—O-optionally substituted heteroaryl.

[0133] Formula (I) contains the substituent R.sup.1. In certain embodiments, R.sup.1 is hydrogen. In certain embodiments, R.sup.1 is optionally substituted acyl. In certain embodiments, R.sup.1 is optionally substituted alkyl (e.g., Me, Et, Pr). In certain embodiments, R.sup.1 is optionally substituted methyl. In certain embodiments, R.sup.1 is unsubstituted methyl. In certain embodiments, R.sup.1 is optionally substituted alkenyl. In certain embodiments, R.sup.1 is optionally substituted alkynyl. In certain embodiments, R.sup.1 is optionally substituted carbocyclyl. In certain embodiments, R.sup.1 is optionally substituted heterocyclyl. In certain embodiments, R.sup.1 is optionally substituted aryl. In certain embodiments, R.sup.1 is optionally substituted heteroaryl. In certain embodiments, R.sup.1 is a nitrogen protecting group.

[0134] In certain embodiments, R.sup.1 is methyl.

[0135] Formula (I) contains the substituents R.sup.2. In certain embodiments, at least one of R.sup.2 is hydrogen. In certain embodiments, at least one instance of R.sup.2 is halogen. In certain embodiments, at least one instance of R.sup.2 is optionally substituted acyl. In certain embodiments, at least one instance of R.sup.2 is optionally substituted alkyl. In certain embodiments, at least one instance of R.sup.2 is optionally substituted alkenyl. In certain embodiments, at least one instance of R.sup.2 is optionally substituted alkynyl. In certain embodiments, at least one instance of R.sup.2 is optionally substituted carbocyclyl. In certain embodiments, at least one instance of R.sup.2 is optionally substituted heterocyclyl. In certain embodiments, at least one instance of R.sup.2 is optionally substituted aryl. In certain embodiments, at least one instance of R.sup.2 is optionally substituted heteroaryl. In certain embodiments, at least one instance of R.sup.2 is —OR.sup.a. In certain embodiments, at least one instance of R.sup.2 is —N(R.sup.a).sub.2.

[0136] In certain embodiments, R.sup.2 is H.

[0137] In certain embodiments, there are m instances of R.sup.2. In certain embodiments, m is 1. In certain embodiments, m is 2. In certain embodiments, m is 3. In certain embodiments, m is 4. In certain embodiments, m is 5. In certain embodiments, m is 6. In certain embodiments, m is 7. In certain embodiments, m is 8. In certain embodiments, m is 9. In certain embodiments, m is 10.

[0138] In certain embodiments, R.sup.2 is H; and m is 1.

[0139] Formula (I) contains the substituents R.sup.3. In certain embodiments, at least one instance of R.sup.3 is hydrogen. In certain embodiments, at least one instance of R.sup.3 is halogen. In certain embodiments, at least one instance of R.sup.3 is optionally substituted acyl. In certain embodiments, at least one instance of R.sup.3 is optionally substituted alkyl. In certain embodiments, at least one instance of R.sup.3 is optionally substituted alkenyl. In certain embodiments, at least one instance of R.sup.3 is optionally substituted alkynyl. In certain embodiments, at least one instance of R.sup.3 is optionally substituted carbocyclyl. In certain embodiments, at least one instance of R.sup.3 is optionally substituted heterocyclyl. In certain embodiments, at least one instance of R.sup.3 is optionally substituted aryl. In certain embodiments, at least one instance of R.sup.3 is optionally substituted heteroaryl. In certain embodiments, at least one instance of R.sup.3 is —OR.sup.b. In certain embodiments, at least one instance of R.sup.3 is —N(R.sup.b).sub.2. In certain embodiments, at least one instance of R.sup.3 is —SR.sup.b.

[0140] In certain embodiments, R.sup.3 is H.

[0141] In certain embodiments, there are n instances of R.sup.3. In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, n is 3. In certain embodiments, n is 4. In certain embodiments, n is 5. In certain embodiments, n is 6. In certain embodiments, n is 7. In certain embodiments, n is 8.

[0142] In certain embodiments, R.sup.3 is H; and n is 1.

[0143] In certain embodiments, R.sup.2 and R.sup.3 are the same. In certain embodiments, R.sup.2 and R.sup.3 are different. In certain embodiments, R.sup.2 is H; and R.sup.3 is H.

[0144] Formula (I) contains the substituent J. In certain embodiments, J is a bond. In certain embodiments, J is alkynylene.

[0145] Formula (I) contains ring A. In certain embodiments, ring A is of the formula:

##STR00011##

In certain embodiments, ring A is of the formula:

##STR00012##

In certain embodiments, ring A is of the formula:

##STR00013##

In certain embodiments, ring A is of the formula:

##STR00014##

In certain embodiments, ring A is of the formula:

##STR00015##

In certain embodiments, ring A is of the formula:

##STR00016##

In certain embodiments, ring A is of the formula:

##STR00017##

In certain embodiments, ring A is of the formula:

##STR00018##

In certain embodiments, ring A is of the formula:

##STR00019##

In certain embodiments, ring A is of the formula:

##STR00020##

In certain embodiments, ring A is of the formula:

##STR00021##

In certain embodiments, ring A is of the formula:

##STR00022##

In certain embodiments, ring A is of the formula:

##STR00023##

In certain embodiments, ring A is of the formula:

##STR00024##

In certain embodiments, ring A is of the formula:

##STR00025##

In certain embodiments, ring A is of the formula:

##STR00026##

In certain embodiments, ring A is of the formula:

##STR00027##

In certain embodiments, J is alkynyl; and ring A is of the formula:

##STR00028##

In certain embodiments, J is alkynyl ring A is of the formula:

##STR00029##

In certain embodiments, J is alkynyl and ring A is of the formula:

##STR00030##

In certain embodiments, J is alkynyl ring A is of the formula:

##STR00031##

In certain embodiments, J is alkynyl and ring A is of the formula:

##STR00032##

[0146] In certain embodiments, Ring A comprises the substituent R.sup.4. In certain embodiments, each instance of R.sup.4 is the same. In certain embodiments, each instance of R.sup.4 is different. In certain embodiments, R.sup.4 is hydrogen. In certain embodiments, R.sup.4 is halogen. In certain embodiments, R.sup.4 is optionally substituted acyl. In certain embodiments, R.sup.4 is optionally substituted alkyl (e.g., Me, Et, Pr). In certain embodiments, R.sup.4 is —CN. In certain embodiments, R.sup.4 is —SCN. In certain embodiments, R.sup.4 is —NO.sub.2. In certain embodiments, R.sup.4 is —C(═O)R.sup.c. In certain embodiments, R.sup.4 is —C(═O)N(R.sup.c).sub.2. In certain embodiments, R.sup.4 is —OR.sup.c. In certain embodiments, R.sup.4 is —N(R.sup.c).sub.2. In certain embodiments, R.sup.4 is —C(═NR.sup.c)R.sup.c. In certain embodiments, R.sup.4 is —C(═NR.sup.c)OR.sup.c. In certain embodiments, R.sup.4 is —C(═NR.sup.c)N(R.sup.c).sub.2. In certain embodiments, R.sup.4 is —C(═O)OR.sup.c. In certain embodiments, R.sup.4 is —NR.sup.cC(═O)R.sup.c. In certain embodiments, R.sup.4 is —NR.sup.cC(═O)OR.sup.c. In certain embodiments, R.sup.4 is —NR.sup.cC(═O)N(R.sup.c).sub.2. In certain embodiments, R.sup.4 is —OC(═O)R.sup.c. In certain embodiments, R.sup.4 is —OC(═O)OR.sup.c. In certain embodiments, R.sup.4 is —OC(═O)N(R.sup.c).sub.2. In certain embodiments, R.sup.4 is —SRC, wherein R.sup.C is independently selected from hydrogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, an oxygen protecting group when attached to an oxygen atom, and a sulfur protecting group when attached to a sulfur atom.

[0147] In certain embodiments, R.sup.4 is H.

[0148] In certain embodiments, R.sup.4 is —C(═O)N(R.sup.c).sub.2. In certain embodiments, R.sup.4 is —C(═O)NH.sub.2.

[0149] In certain embodiments, Ring A comprises the substituent R.sup.5. In certain embodiments, at R.sup.5 is hydrogen. In certain embodiments, R.sup.5 is halogen. In certain embodiments, R.sup.5 is optionally substituted acyl. In certain embodiments, R.sup.5 is optionally substituted alkyl (e.g., Me, Et, Pr). In certain embodiments, R.sup.5 is —CN. In certain embodiments, R.sup.5 is —SCN. In certain embodiments, R.sup.5 is —NO.sub.2. In certain embodiments, R.sup.5 is —C(═O)R.sup.d. In certain embodiments, R.sup.5 is —C(═O)N(R.sup.d).sub.2. In certain embodiments, R.sup.5 is —OR.sup.d. In certain embodiments, R.sup.5 is —N(R.sup.d).sub.2. In certain embodiments, R.sup.5 is —C(═NR.sup.d)R.sup.d. In certain embodiments, R.sup.5 is —C(═NR.sup.d)OR.sup.d. In certain embodiments, R.sup.5 is —C(═NR.sup.d)N(R.sup.d).sub.2. In certain embodiments, R.sup.5 is —C(═O)OR.sup.d. In certain embodiments, R.sup.5 is —NR.sup.dC(═O)R.sup.d. In certain embodiments, R.sup.5 is —NR.sup.dC(═O)OR.sup.d. In certain embodiments, R.sup.5 is —NR.sup.dC(═O)N(R.sup.d).sub.2. In certain embodiments, R.sup.5 is —OC(═O)R.sup.d. In certain embodiments, R.sup.5 is —OC(═O)OR.sup.d. In certain embodiments, R.sup.5 is —OC(═O)N(R.sup.d).sub.2. In certain embodiments, R.sup.5 is —SR.sup.d, wherein R.sup.d is independently selected from hydrogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, an oxygen protecting group when attached to an oxygen atom, and a sulfur protecting group when attached to a sulfur atom.

[0150] In certain embodiments, R.sup.5 is H. In certain embodiments, R.sup.5 is —N(R.sup.d).sub.2. In certain embodiments, R.sup.5 is —NH.sub.2.

[0151] In certain embodiments, R.sup.4 is —C(═O)N(R.sup.d).sub.2; and R.sup.5 is —N(R.sup.d).sub.2. In certain embodiments, R.sup.4 is —C(═O)NH.sub.2; and R.sup.5 is —N(R.sup.d).sub.2. In certain embodiments, R.sup.4 is —C(═O)N(R.sup.c).sub.2; and R.sup.5 is —NH.sub.2. In certain embodiments, R.sup.4 is —C(═O)NH.sub.2; and R.sup.5 is —NH.sub.2.

[0152] In certain embodiments, Ring A comprises the substituent R.sup.6. In certain embodiments, at R.sup.6 is hydrogen. In certain embodiments, R.sup.6 is halogen. In certain embodiments, R.sup.6 is optionally substituted acyl. In certain embodiments, R.sup.6 is optionally substituted alkyl (e.g., Me, Et, Pr). In certain embodiments, R.sup.6 is —CN. In certain embodiments, R.sup.6 is —SCN. In certain embodiments, R.sup.6 is —NO.sub.2. In certain embodiments, R.sup.6 is —C(═O)R.sup.e. In certain embodiments, R.sup.6 is —C(═O)N(R.sup.e).sub.2. In certain embodiments, R.sup.6 is —OR.sup.d. In certain embodiments, R.sup.6 is —N(R.sup.e).sub.2. In certain embodiments, R.sup.6 is —C(═NR.sup.e)R.sup.e. In certain embodiments, R.sup.6 is —C(═NR.sup.e)OR.sup.e. In certain embodiments, R.sup.6 is —C(═NR.sup.e)N(R.sup.e).sub.2. In certain embodiments, R.sup.6 is —C(═O)OR.sup.e. In certain embodiments, R.sup.6 is —NR.sup.eC(═O)R.sup.e. In certain embodiments, R.sup.6 is —NR.sup.eC(═O)OR.sup.e. In certain embodiments, R.sup.6 is —NR.sup.eC(═O)N(R.sup.e).sub.2. In certain embodiments, R.sup.6 is —OC(═O)R.sup.e. In certain embodiments, R.sup.6 is —OC(═O)OR.sup.e. In certain embodiments, R.sup.6 is —OC(═O)N(R.sup.e).sub.2. In certain embodiments, R.sup.6 is —SR.sup.c, wherein R.sup.c is independently selected from hydrogen, optionally substituted acyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, an oxygen protecting group when attached to an oxygen atom, and a sulfur protecting group when attached to a sulfur atom.

[0153] In certain embodiments, R.sup.6 is H.

[0154] In certain embodiments, ring A contains the substituent X.sup.1. In certain embodiments, X.sup.1═N—. In certain embodiments, X.sup.1 is ═C(R.sup.ff)—. In certain embodiments, R.sup.f is hydrogen, halogen, or substituted or unsubstituted C.sub.1-6 alkyl. In certain embodiments, X.sup.1 is ═C(H)—.

[0155] In certain embodiments, ring A contains the substituent X.sup.2. In certain embodiments, X.sup.2═N—. In certain embodiments, X.sup.2 is ═C(R.sup.g)—. In certain embodiments, R.sup.g is hydrogen, halogen, or substituted or unsubstituted C.sub.1-6 alkyl. In certain embodiments, X.sup.2 is ═C(H)—.

[0156] In certain embodiments, ring A contains the substituent X.sup.3. In certain embodiments, X.sup.3═N—. In certain embodiments, X.sup.3 is ═C(R.sup.g)—. In certain embodiments, R.sup.g is hydrogen, halogen, or substituted or unsubstituted C.sub.1-6 alkyl. In certain embodiments, X.sup.2 is ═C(H)—.

[0157] In certain embodiments, ring A contains the substituent X.sup.4. In certain embodiments, X.sup.4═N—. In certain embodiments, X.sup.4 is ═C(R.sup.ff)—. In certain embodiments, R.sup.f is hydrogen, halogen, or substituted or unsubstituted C.sub.1-6 alkyl. In certain embodiments, X.sup.1 is ═C(H)—.

[0158] In certain embodiments, ring A contains the substituent X.sup.4. In certain embodiments, X.sup.4 is ═N—. In certain embodiments, X.sup.4 is ═C(R.sup.1)—. In certain embodiments, X.sup.4 is ═C(H)—. In certain embodiments, R.sup.1 is hydrogen, halogen, or substituted or unsubstituted C.sub.1-6 alkyl (e.g., Me, Et, Pr).

[0159] In certain embodiments, X.sup.4 is ═N—; and R is —CH.sub.2—O-optionally substituted aryl. In certain embodiments, X.sup.4 is ═C(R.sup.ff)—; and R is —CH.sub.2—O-optionally substituted aryl. In certain embodiments, X.sup.4 is ═C(H)—; and R is —CH.sub.2—O-optionally substituted aryl. In certain embodiments, X.sup.4 is ═N—; and R is —CH.sub.2—O-unsubstituted phenyl. In certain embodiments, X.sup.4 is ═C(R.sup.1)—; and R is —CH.sub.2—O-unsubstituted phenyl. In certain embodiments, X.sup.4 is ═C(H)— and R is —CH.sub.2—O-unsubstituted phenyl.

[0160] In certain embodiments, ring A is of the formula:

##STR00033##

In certain embodiments, ring A is of the formula:

##STR00034##

In certain embodiments, ring A is of the formula:

##STR00035##

In certain embodiments, ring A is of the formula:

##STR00036##

In certain embodiments, ring A is of the formula:

##STR00037##

In certain embodiments, ring A is of the formula:

##STR00038##

In certain embodiments, ring A is of the formula:

##STR00039##

In certain embodiments, ring A is of the formula:

##STR00040##

In certain embodiments, ring A is of the formula:

##STR00041##

In certain embodiments, ring A is of the formula:

##STR00042##

In certain embodiments, ring A is of the formula:

##STR00043##

[0161] In certain embodiments, ring A is not of the formula:

##STR00044##

In certain embodiments, ring A is not of the formula:

##STR00045##

In certain embodiments, ring A is not of the formula:

##STR00046##

In certain embodiments, ring A is not of the formula:

##STR00047##

In certain embodiments, ring A is not of the formula:

##STR00048##

In certain embodiments, J is a bond; and ring A is not of the formula:

##STR00049##

In certain embodiments, J is a bond; and ring A is not of the formula:

##STR00050##

In certain embodiments, J is a bond; and ring A is not of the formula:

##STR00051##

In certain embodiments, J is a bond; and ring A is not of the formula:

##STR00052##

[0162] In certain embodiments, a compound of Formula (I) is of the Formula (II):

##STR00053##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

[0163] In certain embodiments, a compound of Formula (I) is of the Formula (III):

##STR00054##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

[0164] In certain embodiments, a compound of Formula (I) is of the Formula (IV):

##STR00055##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

[0165] In certain embodiments, a compound of Formula (I) is of the formula:

##STR00056##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

[0166] In certain embodiments, a compound of Formula (I) is of the formula:

##STR00057##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

[0167] In certain embodiments, a compound of Formula (I) is of the Formula (V):

##STR00058##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

[0168] In certain embodiments, the compound of Formula (V) is of the formula:

##STR00059##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

[0169] In certain embodiments, the compound of Formula (V) is of the formula:

##STR00060##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

[0170] In certain embodiments, a compound of Formula (I) is of the formula:

##STR00061## ##STR00062## ##STR00063## ##STR00064##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

[0171] In certain embodiments, a compound of Formula (I) is of the formula:

##STR00065## ##STR00066## ##STR00067## ##STR00068##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

[0172] In certain embodiments, a compound of Formula (I) is of the Formula (VI):

##STR00069##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

[0173] In certain embodiments, a compound of Formula (I) is of the formula:

##STR00070##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

[0174] In certain embodiments, a compound of Formula (I) is of the formula:

##STR00071## ##STR00072## ##STR00073## ##STR00074##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

[0175] In certain embodiments, a compound of Formula (I) is a compound, or pharmaceutically acceptable salt thereof, of the formula:

TABLE-US-00001 # Structure I-1 [00075] I-2 [00076] I-3 [00077] I-4 [00078] I-5 [00079] I-6 [00080] I-7 [00081] I-8 [00082] I-9 [00083] I-10 [00084]

Pharmaceutical Compositions, Administration, and Kits

[0176] The present disclosure also provides pharmaceutical compositions comprising a compound described herein and optionally a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition further comprises an additional pharmaceutical agent.

[0177] In certain embodiments, the compound described herein is provided in an effective (e.g., effective for inhibiting a kinase, such as BTK, HCK, LYN, BLK, FRK) amount in the pharmaceutical composition. In certain embodiments, the effective amount is a therapeutically effective amount. In certain embodiments, a therapeutically effective amount is an amount effective for inhibiting a kinase. In certain embodiments, a therapeutically effective amount is an amount effective for treating a disease (e.g., a disease associated with aberrant activity of a kinase (e.g., proliferative disease)). In certain embodiments, a therapeutically effective amount is an amount effective for inhibiting the activity of a kinase and treating a disease (e.g., a disease associated with aberrant activity of a kinase (e.g., proliferative disease (e.g., cancer (e.g., breast cancer, colon cancer, testicular cancer, CNS cancer, stomach cancer, lymphoma (e.g., B-cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgM secreting lymphoplasmacytic lymphoma (i.e., Waldenström'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, myelogenous leukemia (e.g., chronic myelogenous leukemia, acute myelogenous leukemia))))).

[0178] In certain embodiments, the effective amount is an amount effective for inhibiting the activity of a kinase 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, the effective amount is an amount effective for inhibiting the activity of a kinase by not more than 10%, not more than 20%, not more than 30%, not more than 40%, not more than 50%, not more than 60%, not more than 70%, not more than 80%, not more than 90%, not more than 95%, or not more than 98%.

[0179] In certain embodiments, the subject is an animal. The animal may be of either sex and may be at any stage of development. In certain embodiments, the subject described herein is a human (e.g., an adult, juvenile, or 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 domesticated animal, such as a dog, cat, cow, pig, horse, sheep, or goat. In certain embodiments, the subject is a dog. In certain embodiments, the subject is a companion animal, such as a dog or cat. In certain embodiments, the subject is a livestock animal, such as a cow, pig, horse, sheep, or 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., mouse, rat), dog, pig, or non-human primate. In certain embodiments, the subject is a genetically engineered animal. In certain embodiments, the subject is a transgenic animal (e.g., transgenic mice, transgenic pigs). In certain embodiments, the subject is a fish or reptile.

[0180] In certain embodiments, the biological sample or cell (e.g., the biological sample or cell being contacted with a compound or pharmaceutical composition described herein) is in vitro. In certain embodiments, the biological sample or cell is in vivo or ex vivo. In certain embodiments, the cell is a malignant cell or premalignant cell.

[0181] Pharmaceutical compositions described herein can be prepared by any method known in the art of pharmacology. In general, such preparatory methods include bringing the compound described herein (i.e., the “active ingredient”) into association with a carrier or excipient, and/or one or more other accessory ingredients, and then, if necessary and/or desirable, shaping, and/or packaging the product into a desired single- or multi-dose unit.

[0182] Pharmaceutical compositions can be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses. A “unit dose” is a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject and/or a convenient fraction of such a dosage, such as one-half or one-third of such a dosage.

[0183] Relative amounts of the active ingredient, the pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition described herein will vary, depending upon the identity, size, and/or condition of the subject treated and further depending upon the route by which the composition is to be administered. The composition may comprise between 0.1% and 100% (w/w) active ingredient.

[0184] Pharmaceutically acceptable excipients used in the manufacture of provided pharmaceutical compositions include inert diluents, dispersing and/or granulating agents, surface active agents and/or emulsifiers, disintegrating agents, binding agents, preservatives, buffering agents, lubricating agents, and/or oils. Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening, flavoring, and perfuming agents may also be present in the composition.

[0185] Exemplary diluents include calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, cornstarch, powdered sugar, and mixtures thereof.

[0186] 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 sponge, 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), methylcellulose, 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.

[0187] Exemplary surface active agents and/or emulsifiers include natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, wax, 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, triacetin monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxy polymethylene, polyacrylic acid, acrylic acid polymer, and carboxyvinyl polymer), carrageenan, cellulosic derivatives (e.g., carboxymethylcellulose sodium, powdered cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate (Tween® 20), polyoxyethylene sorbitan (Tween® 60), polyoxyethylene sorbitan monooleate (Tween® 80), sorbitan monopalmitate (Span® 40), sorbitan monostearate (Span® 60), sorbitan tristearate (Span® 65), glyceryl monooleate, sorbitan monooleate (Span® 80), polyoxyethylene esters (e.g., polyoxyethylene monostearate (Myrj® 45), polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate, and Solutol*), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g., Cremophor®), polyoxyethylene ethers, (e.g., polyoxyethylene lauryl ether (Brij® 30)), poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, Pluronic® F-68, poloxamer P-188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium, and/or mixtures thereof.

[0188] Exemplary binding agents include starch (e.g., cornstarch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums (e.g., acacia, sodium alginate, extract of Irish moss, panwar gum, ghatti gum, mucilage of isapol husks, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate (Veegum©), and larch arabogalactan), alginates, polyethylene oxide, polyethylene glycol, inorganic calcium salts, silicic acid, polymethacrylates, waxes, water, alcohol, and/or mixtures thereof.

[0189] Exemplary preservatives include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, antiprotozoan 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.

[0190] 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.

[0191] 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, and the like), 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, ethyl alcohol, glycerin, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and thimerosal.

[0192] Exemplary antifungal preservatives include butyl paraben, methyl paraben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid.

[0193] Exemplary alcohol preservatives include ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate, and phenylethyl alcohol.

[0194] 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.

[0195] Other preservatives include tocopherol, tocopherol acetate, deteroxime mesylate, cetrimide, butylated hydroxyanisol (BHA), butylated hydroxytoluened (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, Glydant® Plus, Phenonip®, methylparaben, Germall® 115, Germaben® II, Neolone®, Kathon®, and Euxyl®.

[0196] Exemplary buffering agents include citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic sodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, and mixtures thereof.

[0197] Exemplary lubricating agents include magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behanate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and mixtures thereof.

[0198] Exemplary natural oils include almond, apricot kernel, avocado, babassu, bergamot, black current seed, borage, cade, camomile, canola, caraway, carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cotton seed, emu, eucalyptus, evening primrose, fish, flaxseed, geraniol, gourd, grape seed, hazel nut, hyssop, isopropyl myristate, jojoba, kukui nut, lavandin, lavender, lemon, Litsea cubeba, macademia 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, savoury, sea buckthorn, sesame, shea butter, silicone, soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut, and wheat germ oils. 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.

[0199] Liquid dosage forms for oral and parenteral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredients, the liquid dosage forms may comprise inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents. In certain embodiments for parenteral administration, the conjugates described herein are mixed with solubilizing agents such as Cremophor®, alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and mixtures thereof.

[0200] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions can be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can be a sterile injectable solution, suspension, or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution, U.S.P., and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or di-glycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

[0201] The injectable formulations can be sterilized, for example, by filtration through a bacterial-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.

[0202] In order to prolong the effect of a drug, it is often desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility.

[0203] The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form may be accomplished by dissolving or suspending the drug in an oil vehicle.

[0204] 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.

[0205] 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 starches, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, (c) humectants such as glycerol, (d) disintegrating agents such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, (e) solution retarding agents such as paraffin, (f) absorption accelerators such as quaternary ammonium compounds, (g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, (h) absorbents such as kaolin and bentonite clay, and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may include a buffering agent.

[0206] Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the art of pharmacology. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of encapsulating compositions which can be used include polymeric substances and waxes. Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

[0207] The active ingredient can be in a micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings, and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active ingredient can be admixed with at least one inert diluent such as sucrose, lactose, or starch. Such dosage forms may comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may comprise buffering agents. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of encapsulating agents which can be used include polymeric substances and waxes.

[0208] Dosage forms for topical and/or transdermal administration of a compound described herein may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, and/or patches. Generally, the active ingredient is admixed under sterile conditions with a pharmaceutically acceptable carrier or excipient and/or any needed preservatives and/or buffers as can be required. Additionally, the present disclosure contemplates the use of transdermal patches, which often have the added advantage of providing controlled delivery of an active ingredient to the body. Such dosage forms can be prepared, for example, by dissolving and/or dispensing the active ingredient in the proper medium. Alternatively or additionally, the rate can be controlled by either providing a rate controlling membrane and/or by dispersing the active ingredient in a polymer matrix and/or gel.

[0209] Suitable devices for use in delivering intradermal pharmaceutical compositions described herein include short needle devices. Intradermal compositions can be administered by devices which limit the effective penetration length of a needle into the skin. Alternatively or additionally, conventional syringes can be used in the classical mantoux method of intradermal administration. Jet injection devices which deliver liquid formulations to the dermis via a liquid jet injector and/or via a needle which pierces the stratum corneum and produces a jet which reaches the dermis are suitable. Ballistic powder/particle delivery devices which use compressed gas to accelerate the compound in powder form through the outer layers of the skin to the dermis are suitable.

[0210] Formulations 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 formulations may, for example, comprise from about 1% to about 10% (w/w) active ingredient, although the concentration of the active ingredient can be as high as the solubility limit of the active ingredient in the solvent. Formulations for topical administration may further comprise one or more of the additional ingredients described herein.

[0211] A pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation suitable for pulmonary administration via the buccal cavity. Such a formulation may comprise dry particles which comprise the active ingredient and which have a diameter in the range from about 0.5 to about 7 nanometers, or from about 1 to about 6 nanometers. Such compositions are conveniently in the form of dry powders for administration using a device comprising a dry powder reservoir to which a stream of propellant can be directed to disperse the powder and/or using a self-propelling solvent/powder dispensing container such as a device comprising the active ingredient dissolved and/or suspended in a low-boiling propellant in a sealed container. Such powders comprise particles wherein at least 98% of the particles by weight have a diameter greater than 0.5 nanometers and at least 95% of the particles by number have a diameter less than 7 nanometers. Alternatively, at least 95% of the particles by weight have a diameter greater than 1 nanometer and at least 90% of the particles by number have a diameter less than 6 nanometers. Dry powder compositions may include a solid fine powder diluent such as sugar and are conveniently provided in a unit dose form.

[0212] Low boiling propellants generally include liquid propellants having a boiling point of below 65° F. at atmospheric pressure. Generally the propellant may constitute 50 to 99.9% (w/w) of the composition, and the active ingredient may constitute 0.1 to 20% (w/w) of the composition. The propellant may further comprise additional ingredients such as a liquid non-ionic and/or solid anionic surfactant and/or a solid diluent (which may have a particle size of the same order as particles comprising the active ingredient).

[0213] Pharmaceutical compositions described herein formulated for pulmonary delivery may provide the active ingredient in the form of droplets of a solution and/or suspension. Such formulations can be prepared, packaged, and/or sold as aqueous and/or dilute alcoholic solutions and/or suspensions, optionally sterile, comprising the active ingredient, and may conveniently be administered using any nebulization and/or atomization device. Such formulations may further comprise one or more additional ingredients including, but not limited to, a flavoring agent such as saccharin sodium, a volatile oil, a buffering agent, a surface active agent, and/or a preservative such as methylhydroxybenzoate. The droplets provided by this route of administration may have an average diameter in the range from about 0.1 to about 200 nanometers.

[0214] Formulations described herein as being useful for pulmonary delivery are useful for intranasal delivery of a pharmaceutical composition described herein. Another formulation suitable for intranasal administration is a coarse powder comprising the active ingredient and having an average particle from about 0.2 to 500 micrometers. Such a formulation is administered by rapid inhalation through the nasal passage from a container of the powder held close to the nares.

[0215] Formulations for nasal administration may, for example, comprise from about as little as 0.1% (w/w) to as much as 100% (w/w) of the active ingredient, and may comprise one or more of the additional ingredients described herein. A pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation for buccal administration. Such formulations may, for example, be in the form of tablets and/or lozenges made using conventional methods, and may contain, for example, 0.1 to 20% (w/w) active ingredient, the balance comprising an orally dissolvable and/or degradable composition and, optionally, one or more of the additional ingredients described herein. Alternately, formulations for buccal administration may comprise a powder and/or an aerosolized and/or atomized solution and/or suspension comprising the active ingredient. Such powdered, aerosolized, and/or aerosolized formulations, when dispersed, may have an average particle and/or droplet size in the range from about 0.1 to about 200 nanometers, and may further comprise one or more of the additional ingredients described herein.

[0216] A pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation for ophthalmic administration. Such formulations may, for example, be in the form of eye drops including, for example, a 0.1-1.0% (w/w) solution and/or suspension of the active ingredient in an aqueous or oily liquid carrier or excipient. Such drops may further comprise buffering agents, salts, and/or one or more other of the additional ingredients described herein. Other ophthalmically-administrable formulations which are useful include those which comprise the active ingredient in microcrystalline form and/or in a liposomal preparation. Ear drops and/or eye drops are also contemplated as being within the scope of this disclosure.

[0217] Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and/or perform such modification with ordinary experimentation.

[0218] Compounds provided herein are typically formulated in dosage unit form for ease of administration and uniformity of dosage. It will be understood, however, that the total daily usage of the compositions described herein will be decided by a physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject or organism will depend upon a variety of factors including the disease being treated and the severity of the disorder; the activity of the specific active ingredient employed; the specific composition employed; the age, body weight, general health, sex, and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific active ingredient employed; the duration of the treatment; drugs used in combination or coincidental with the specific active ingredient employed; and like factors well known in the medical arts.

[0219] The compounds and compositions provided herein can be administered by any route, including enteral (e.g., oral), parenteral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, subcutaneous, intraventricular, transdermal, interdermal, rectal, intravaginal, intraperitoneal, topical (as by powders, ointments, creams, and/or drops), mucosal, nasal, bucal, sublingual; by intratracheal instillation, bronchial instillation, and/or inhalation; and/or as an oral spray, nasal spray, and/or aerosol. Specifically contemplated routes are oral administration, intravenous administration (e.g., systemic intravenous injection), regional administration via blood and/or lymph supply, and/or direct administration to an affected site. In general, the most appropriate route of administration will depend upon a variety of factors including the nature of the agent (e.g., its stability in the environment of the gastrointestinal tract), and/or the condition of the subject (e.g., whether the subject is able to tolerate oral administration). In certain embodiments, the compound or pharmaceutical composition described herein is suitable for topical administration to the eye of a subject.

[0220] The exact amount of a compound required to achieve an effective amount will vary from subject to subject, depending, for example, on species, age, and general condition of a subject, severity of the side effects or disorder, identity of the particular compound, mode of administration, and the like. An effective amount may be included in a single dose (e.g., single oral dose) or multiple doses (e.g., multiple oral doses). In certain embodiments, when multiple doses are administered to a subject or applied to a biological sample or cell, any two doses of the multiple doses include different or substantially the same amounts of a compound described herein. In certain embodiments, when multiple doses are administered to a subject or applied to a biological sample or cell, the frequency of administering the multiple doses to the subject or applying the multiple doses to the biological sample or cell is three doses a day, two doses a day, one dose a day, one dose every other day, one dose every third day, one dose every week, one dose every two weeks, one dose every three weeks, or one dose every four weeks. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the biological sample or cell is one dose per day. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the biological sample or cell is two doses per day. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the biological sample or cell is three doses per day. In certain embodiments, when multiple doses are administered to a subject or applied to a biological sample or cell, the duration between the first dose and 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 lifetime of the subject or cell. In certain embodiments, the duration between the first dose and last dose of the multiple doses is three months, six months, or one year. In certain embodiments, the duration between the first dose and last dose of the multiple doses is the lifetime of the subject or cell. In certain embodiments, a dose (e.g., a single dose, or any dose of multiple doses) described herein includes independently between 0.1 μg and 1 μg, between 0.001 mg and 0.01 mg, between 0.01 mg and 0.1 mg, between 0.1 mg and 1 mg, between 1 mg and 3 mg, between 3 mg and 10 mg, between 10 mg and 30 mg, between 30 mg and 100 mg, between 100 mg and 300 mg, between 300 mg and 1,000 mg, or between 1 g and 10 g, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 1 mg and 3 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 3 mg and 10 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 10 mg and 30 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 30 mg and 100 mg, inclusive, of a compound described herein.

[0221] Dose ranges as described herein provide guidance for the administration of provided pharmaceutical compositions to an adult. The amount to be administered to, for example, a child or an adolescent can be determined by a medical practitioner or person skilled in the art and can be lower or the same as that administered to an adult.

[0222] A compound or composition, as described herein, can be administered in combination with one or more additional pharmaceutical agents (e.g., therapeutically and/or prophylactically active agents). The compounds or compositions can be administered in combination with additional pharmaceutical agents that improve their activity (e.g., activity (e.g., potency and/or efficacy) in treating a disease in a subject in need thereof, in preventing a disease in a subject in need thereof, in inhibiting the activity of a kinase (e.g., BTK, HCK, LYN, BLK, FRK) in a subject, biological sample, or cell), improve bioavailability, improve safety, reduce drug resistance, reduce and/or modify metabolism, inhibit excretion, and/or modify distribution in a subject, biological sample, or cell. It will also be appreciated that the therapy employed may achieve a desired effect for the same disorder, and/or it may achieve different effects. In certain embodiments, a pharmaceutical composition described herein including a compound described herein and an additional pharmaceutical agent shows a synergistic effect that is absent in a pharmaceutical composition including one of the compound and the additional pharmaceutical agent, but not both.

[0223] The compound or composition can be administered concurrently with, prior to, or subsequent to one or more additional pharmaceutical agents, which may be useful as, e.g., combination therapies. Pharmaceutical agents include therapeutically active agents. Pharmaceutical agents also include prophylactically active agents. Pharmaceutical agents include small organic molecules such as drug compounds (e.g., compounds approved for human or veterinary use by the U.S. Food and Drug Administration as provided in the Code of Federal Regulations (CFR)), peptides, proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides or proteins, small molecules linked to proteins, glycoproteins, steroids, nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins, and cells. In certain embodiments, the additional pharmaceutical agent is a pharmaceutical agent useful for treating and/or preventing a disease (e.g., proliferative disease, cancer, inflammatory disease, autoimmune disease, genetic disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder) or premalignant condition. Each additional pharmaceutical agent may be administered at a dose and/or on a time schedule determined for that pharmaceutical agent. The additional pharmaceutical agents may also be administered together with each other and/or with the compound or composition described herein in a single dose or administered separately in different doses. The particular combination to employ in a regimen will take into account compatibility of the compound described herein with the additional pharmaceutical agent(s) and/or the desired therapeutic and/or prophylactic effect to be achieved. In general, it is expected that the additional pharmaceutical agent(s) in combination be utilized at levels that do not exceed the levels at which they are utilized individually. In some embodiments, the levels utilized in combination will be lower than those utilized individually.

[0224] The additional pharmaceutical agents include, but are not limited to, cytotoxic chemotherapeutic agents, epigenetic modifiers, glucocorticoids, immunotherapeutic agents, anti-proliferative agents, anti-cancer agents, anti-angiogenesis agents, anti-inflammatory agents, immunosuppressants, anti-bacterial agents, anti-viral agents, cardiovascular agents, cholesterol-lowering agents, anti-diabetic agents, anti-allergic agents, contraceptive agents, pain-relieving agents, and a combination thereof. In certain embodiments, the additional pharmaceutical agent is an anti-proliferative agent (e.g., anti-cancer agent). In certain embodiments, the additional pharmaceutical agent is ABITREXATE (methotrexate), ADE, Adriamycin 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 (obinutuzumab), GLEEVEC (imatinib mesylate), Hyper-CVAD, ICLUSIG (ponatinib hydrochloride), IMBRUVICA (ibrutinib), LEUKERAN (chlorambucil), LINFOLIZIN (chlorambucil), MARQIBO (vincristine sulfate liposome), METHOTREXATE LPF (methorexate), MEXATE (methotrexate), MEXATE-AQ (methotrexate), mitoxantrone hydrochloride, MUSTARGEN (mechlorethamine hydrochloride), MYLERAN (busulfan), NEOSAR (cyclophosphamide), ONCASPAR (Pegaspargase), PURINETHOL (mercaptopurine), PURIXAN (mercaptopurine), Rubidomycin (daunorubicin hydrochloride), SPRYCEL (dasatinib), SYNRIBO (omacetaxine mepesuccinate), 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 pharmaceutical agent is an anti-lymphoma agent. In certain embodiments, the additional pharmaceutical agent is ABITREXATE (methotrexate), ABVD, ABVE, ABVE-PC, ADCETRIS (brentuximab vedotin), ADRIAMYCIN PFS (doxorubicin hydrochloride), ADRIAMYCIN RDF (doxorubicin hydrochloride), AMBOCHLORIN (chlorambucil), AMBOCLORIN (chlorambucil), ARRANON (nelarabine), BEACOPP, BECENUM (carmazine), BELEODAQ (belinostat), BEXXAR (tositumomab and iodine 1131 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), Lomustine, MATULANE (procarbazine hydrochloride), METHOTREXATE LPF (methotrexate), MEXATE (methotrexate), MEXATE-AQ (methotrexate), MOPP, MOZOBIL (plerixafor), MUSTARGEN (mechlorethamine hydrochloride), NEOSAR (cyclophosphamide), OEPA, ONTAK (denileukin diftitox), 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 pharmaceutical agent is REVLIMID (lenalidomide), DACOGEN (decitabine), VIDAZA (azacitidine), CYTOSAR-U (cytarabine), IDAMYCIN (idarubicin), CERUBIDINE (daunorubicin), LEUKERAN (chlorambucil), NEOSAR (cyclophosphamide), FLUDARA (fludarabine), LEUSTATIN (cladribine), ABITREXATE (methotrexate), ABRAXANE (paclitaxel albumin-stabilized nanoparticle formulation), AC, AC-T, ADE, ADRIAMYCIN PFS (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 (lomustine), CERUBIDINE (daunorubicin hydrochloride), CERVARIX (recombinant HPV bivalent vaccine), CLAFEN (cyclophosphamide), CMF, COMETRIQ (cabozantinib-s-malate), COSMEGEN (dactinomycin), CYFOS (ifosfamide), CYRAMZA (ramucirumab), CYTOSAR-U (cytarabine), CYTOXAN (cyclophosphamide), DACOGEN (decitabine), DEGARELIX, DOXIL (doxorubicin hydrochloride liposome), DOXORUBICIN HYDROCHLORIDE, DOX-SL (doxorubicin hydrochloride liposome), DTIC-DOME (dacarbazine), EFUDEX (fluorouracil), ELLENCE (epirubicin hydrochloride), ELOXATIN (oxaliplatin), ERBITUX (cetuximab), ERIVEDGE (vismodegib), ETOPOPHOS (etoposide phosphate), EVACET (doxorubicin hydrochloride liposome), FARESTON (toremifene), FASLODEX (fulvestrant), FEC, FEMARA (letrozole), FLUOROPLEX (fluorouracil), FOLEX (methotrexate), FOLEX 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 dimaleate), GLEEVEC (imatinib mesylate), GLIADEL (carmustine implant), GLIADEL WAFER (carmustine implant), HERCEPTIN (trastuzumab), HYCAMTIN (topotecan hydrochloride), IFEX (ifosfamide), IFOSFAMIDUM (ifosfamide), INLYTA (axitinib), INTRON A (recombinant interferon alfa-2b), IRESSA (gefitinib), IXEMPRA (ixabepilone), JAKAFI (ruxolitinib phosphate), JEVTANA (cabazitaxel), KADCYLA (ado-trastuzumab emtansine), KEYTRUDA (pembrolizumab), KYPROLIS (carfilzomib), LIPODOX (doxorubicin hydrochloride liposome), LUPRON (leuprolide acetate), LUPRON DEPOT (leuprolide acetate), LUPRON DEPOT-3 MONTH (leuprolide acetate), LUPRON DEPOT-4 MONTH (leuprolide acetate), LUPRON DEPOT-PED (leuprolide acetate), MEGACE (megestrol acetate), MEKINIST (trametinib), METHAZOLASTONE (temozolomide), METHOTREXATE LPF (methotrexate), MEXATE (methotrexate), MEXATE-AQ (methotrexate), MITOXANTRONE HYDROCHLORIDE, 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 (peginterferon alfa-2b), PEMETREXED DISODIUM, 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 (regorafenib), 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), VINCASAR PFS (vincristine sulfate), VOTRIENT (pazopanib hydrochloride), WELLCOVORIN (leucovorin calcium), XALKORI (crizotinib), XELODA (capecitabine), XELOX, XGEVA (denosumab), XOFIGO (radium 223 dichloride), XTANDI (enzalutamide), YERVOY (ipilimumab), ZALTRAP (ziv-aflibercept), ZELBORAF (vemurafenib), ZOLADEX (goserelin acetate), ZOMETA (zoledronic acid), ZYKADIA (ceritinib), ZYTIGA (abiraterone acetate), ENMD-2076, PCI-32765, AC220, dovitinib lactate (TK1258, CHIR-258), BIBW 2992 (TOVOK™), SGX523, PF-04217903, PF-02341066, PF-299804, BMS-777607, ABT-869, MP470, BIBF 1120 (VARGATEF®), 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, temsirolimus (CCI-779), everolimus (RAD-001), ridaforolimus, AP23573 (Ariad), AZD8055, BEZ235, BGT226, XL765, PF-4691502, GDC0980, SF1126, and OSI-027), oblimersen, gemcitabine, carminomycin, leucovorin, pemetrexed, cyclophosphamide, dacarbazine, procarbizine, prednisolone, dexamethasone, campathecin, plicamycin, asparaginase, aminopterin, methopterin, porfiromycin, melphalan, leurosidine, leurosine, chlorambucil, trabectedin, procarbazine, discodermolide, carminomycin, aminopterin, and hexamethyl melamine, or a combination thereof. In certain embodiments, the additional pharmaceutical agent is a cytotoxic chemotherapeutic agent (e.g., gemcitabine, cytarabine, daunorubicin, doxorubicin, vincristine, 1-asparaginase, cyclophosphamide, or etoposide). In certain embodiments, the additional pharmaceutical agent is an epigenetic modifier such as azacitidine or romidepsin. In certain embodiments, the additional pharmaceutical agent is ruxolitinib, BBT594, CHZ868, CYT387, or BMS911543. In certain embodiments, the additional pharmaceutical agent is an inhibitor of a tyrosine kinase. In some embodiments, the additional pharmaceutical agent is a topoisomerase inhibitor, a MCL1 inhibitor, a BCL-2 inhibitor, a BCL-xL inhibitor, a BRD4 inhibitor, a BRCA1 inhibitor, BRCA2 inhibitor, HER1 inhibitor, HER2 inhibitor, a CDK9 inhibitor, a Jumonji histone demethylase inhibitor, or a DNA damage inducer. In certain embodiments, the additional pharmaceutical agent is a BCL-2 inhibitor. In certain embodiments, the additional pharmaceutical agent is venetoclax. In some embodiments, the additional pharmaceutical agent is etoposide, obatoclax, navitoclax, JQ1, 4-(((5′-chloro-2′-(((1R,4R)-4-(((R)-1-methoxypropan-2-yl)amino)cyclohexyl)amino)-[2,4′-bipyridin]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile, JIB04, or cisplatin. In certain embodiments, the additional pharmaceutical agent is a binder or inhibitor of a kinase (e.g., JAK, ABL1, CDC2L1, CDC2L2, CSF1R, DDR1, DDR2, FLT4, KIT, PDGFRB, RET, TAOK2, or a combination thereof). In certain embodiments, the additional pharmaceutical agent is an antibody or a fragment thereof (e.g., monoclonal antibody). In certain embodiments, the additional pharmaceutical agent is a tyrosine kinase inhibitor. In certain embodiments, the additional pharmaceutical agent is selected from the group consisting of epigenetic or transcriptional modulators (e.g., DNA methyltransferase inhibitors, histone deacetylase inhibitors (HDAC inhibitors), lysine methyltransferase inhibitors), antimitotic 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), modulators of protein stability (e.g., proteasome inhibitors), Hsp90 inhibitors, glucocorticoids, all-trans retinoic acids, and other agents that promote differentiation. In certain embodiments, the additional pharmaceutical 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 pharmaceutical agent is an immunomodulator. In certain embodiments, the additional pharmaceutical agent is an immune checkpoint inhibitor. In certain embodiments, the additional pharmaceutical agent is a programmed cell death 1 protein (PD-1) inhibitor. In certain embodiments, the additional pharmaceutical agent is a programmed cell death 1 protein ligand 1 (PD-L1) inhibitor. In certain embodiments, the additional pharmaceutical agent is a cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) inhibitor. In certain embodiments, the additional pharmaceutical agent is a BTK inhibitor (e.g., ibrutinib, CC-292, ONO-4059, evobrutinib, spebrutinib, BGB-3111, HM71224, or ACP-196 (i.e., acalabrutinib)). In certain embodiments, the compounds described herein or pharmaceutical compositions can be administered in combination with an anti-cancer therapy including, but not limited to, surgery, radiation therapy, and transplantation (e.g., stem cell transplantation, bone marrow transplantation).

[0225] In certain embodiments, the additional pharmaceutical agent is a BCL-2 inhibitor (e.g., venetoclax, navitoclax, obatoclax), a BCL-2/BCL-xL inhibitor (e.g., APG-1252, BM-1197).

[0226] In certain embodiments, the additional pharmaceutical agent is venetoclax.

[0227] Also encompassed by the present disclosure are kits (e.g., pharmaceutical packs). In certain embodiments, the kit comprises a compound or pharmaceutical composition described herein, and instructions for using the compound or pharmaceutical composition. In certain embodiments, the kit comprises a first container, wherein the first container includes the compound or pharmaceutical composition. In some embodiments, the kit further comprises a second container. In certain embodiments, the second container includes an excipient (e.g., an excipient for dilution or suspension of the compound or pharmaceutical composition). In certain embodiments, the second container includes an additional pharmaceutical agent. In some embodiments, the kit further comprises a third container. In certain embodiments, the third container includes an additional pharmaceutical agent. In some embodiments, the compound or pharmaceutical composition included in the first container and the excipient or additional pharmaceutical agent included in the second container are combined to form one unit dosage form. In some embodiments, the compound or pharmaceutical composition included in the first container, the excipient included in the second container, and the additional pharmaceutical agent included in the third container are combined to form one unit dosage form. In certain embodiments, each of the first, second, and third containers is independently a vial, ampule, bottle, syringe, dispenser package, tube, or inhaler.

[0228] In certain embodiments, the instructions are for administering the compound or pharmaceutical composition to a subject (e.g., a subject in need of treatment or prevention of a disease described herein). In certain embodiments, the instructions are for contacting a biological sample or cell with the compound or pharmaceutical composition. In certain embodiments, the instructions comprise information required by a regulatory agency, such as the U.S. Food and Drug Administration (FDA) or the European Agency for the Evaluation of Medicinal Products (EMA). In certain embodiments, the instructions comprise prescribing information.

[0229] The compounds, pharmaceutical compositions, and kits described herein may synergistically augment inhibition of a kinase (e.g., BTK, HCK, LYN, BLK, FRK) induced by the additional pharmaceutical agent(s) in the biological sample or subject. Thus, the combination of the compounds, pharmaceutical compositions, or kits with additional pharmaceutical agent(s) may be useful in treating diseases resistant to a treatment using the additional pharmaceutical agent(s) without the compounds, pharmaceutical compositions, or kits described herein.

Methods of Treatment and Uses

[0230] The present disclosure provides methods of modulating (e.g., inhibiting or increasing) the activity (e.g., aberrant activity, such as increased or decreased activity) of a kinase (e.g., BTK, HCK, LYN). The present disclosure also provides methods of modulating (e.g., inhibiting or increasing) the activity (e.g., undesired or aberrant activity, such as increased activity (e.g., activity above normal levels) or decreased activity (e.g., activity below normal levels)), of a kinase in a subject, biological sample, or cell. The present disclosure also provides methods for the treatment of a range of diseases and conditions, such as diseases and conditions associated with undesired or aberrant activity (e.g., increased activity) or overexpression of a kinase (e.g., BTK, HCK, LYN). In certain embodiments, the diseases include a proliferative disease (e.g., an IgM gammopathy (e.g., an 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, stomach cancer, lymphoma (e.g., B-cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgM secreting lymphoplasmacytic lymphoma (i.e., Waldenström'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 myelomas (e.g., IgM multiple myeloma)), and leukemia (e.g., chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia, myelogenous leukemia (e.g., chronic myelogenous leukemia, acute myelogenous leukemia (e.g., mast cell leukemia), myeloproliferative diseases (e.g., myelodysplastic syndrome))))).

[0231] In another aspect, the present disclosure provides methods of treating a disease in a subject in need thereof, the method comprising administering to the subject in need thereof an effective amount (e.g., therapeutically effective amount) of a compound described herein or a pharmaceutical composition described herein.

[0232] In another aspect, the present disclosure provides methods of preventing a disease in a subject in need thereof, the method comprising administering to the subject in need thereof an effective amount (e.g., prophylactically effective amount) of a compound described herein or a pharmaceutical composition described herein.

[0233] In another aspect, the present disclosure provides methods of inhibiting the activity of a kinase 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.

[0234] In another aspect, the present disclosure provides methods of inhibiting the activity of a kinase in a biological sample (e.g., an in vitro biological sample), the method comprising contacting the biological sample with an effective amount of a compound described herein or a pharmaceutical composition described herein.

[0235] In another aspect, the present disclosure provides methods of inhibiting the activity of a kinase in a cell (e.g., an in vitro cell), the method comprising contacting the cell with an effective amount of a compound described herein or a pharmaceutical composition described herein.

[0236] Without wishing to be bound by any particular theory, in certain embodiments the compounds described herein are able to bind (e.g., covalently modify) the kinase being inhibited. In certain embodiments, a compound described herein is able to bind (e.g., covalently modify) to the kinase. In certain embodiments, the kinase is HCK. In certain embodiments, the kinase is BTK. In certain embodiments, the kinase is LYN.

[0237] In certain embodiments, provided are methods of decreasing the activity of a kinase (e.g., HCK, BTK, LYN) in a subject, biological sample, or cell 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 a kinase in a subject, biological sample, or cell is decreased 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 a kinase in a subject, biological sample, or cell is selectively inhibited by the method. In some embodiments, the activity of a kinase (e.g., HCK, BTK, LYN) in a subject, biological sample, or cell is selectively decreased by a compound or pharmaceutical composition described herein.

[0238] A disease, including proliferative disease, may be associated with aberrant or undesired activity of a kinase, and/or overexpression of the kinase. Aberrant or undesired activity of a kinase may be an increased or a decreased level of activity of the kinase. Proliferative diseases are sometimes associated with abnormal levels of HCK, BTK, or LYN activity, frequently through increased or decreased HCK, BTK, or LYN activation. Inhibition of the activity of HCK, BTK, or LYN would be expected to inhibit phosphorylation. In certain embodiments, HCK, BTK, or LYN is not overexpressed, but the activity of HCK, BTK, or LYN is increased. In certain embodiments, HCK, BTK, or LYN is overexpressed, and the activity of HCK, BTK, or LYN is increased. The compounds and pharmaceutical compositions described herein may inhibit the activity of HCK, BTK, or LYN and be useful in treating and/or preventing diseases, such as diseases associated with the aberrant, increased, or undesired activity of a kinase, over activation of the kinase, and/or overexpression of the kinase.

[0239] In certain embodiments, the disease (e.g., the disease to be treated or prevented by a method described herein) is associated with the increased activity of a kinase (e.g., HCK, BTK, LYN). In certain embodiments, the disease is associated with overexpression of a kinase (e.g., HCK, BTK, LYN). In certain embodiments, the disease is a proliferative disease. In certain embodiments, the proliferative disease is cancer. In certain embodiments, the cancer is associated with a mutation in MYD88. In another embodiment, the cancer is associated with mutated BTK. In certain embodiments, the proliferative disease is mastocytosis. In certain embodiments, the mastocytosis is systemic mastocytosis. In certain embodiments, the proliferative disease is an IgM gammopathy. In certain embodiments, the IgM gammopathy is IgM monoclonal gammopathy with undetermined significance.

[0240] In certain embodiments, the cancer is breast cancer. In certain embodiments, the cancer is colon cancer. In certain embodiments, the cancer is testicular cancer. In certain embodiments, the cancer is CNS cancer. In certain embodiments, the cancer is stomach cancer. In certain embodiments, the cancer is lymphoma. In certain embodiments, the lymphoma is a B-cell Lymphoma. In certain embodiments, the B-cell lymphoma is lymphoplasmacytic lymphoma. In certain embodiments, the lymphoplasmacytic lymphoma is IgM secreting lymphoplasmacytic lymphoma (i.e., Waldenström's macroglobulinemia). In certain embodiments, the lymphoplasmacytic lymphoma is non-IgM secreting. In certain embodiments, the B-cell lymphoma is Diffuse Large B-Cell Lymphoma (DLBCL). In certain embodiments, the DLBCL is activated B-cell-like (ABC)-DLBCL. In certain embodiments, the 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 myelogenous leukemia. In certain embodiments, the myelogenous leukemia is chronic myelogenous leukemia. In certain embodiments, the myelogenous leukemia is acute myelogenous leukemia. In certain embodiments, the acute myelogenous 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.

[0241] Further provided herein is a method of treating a subject comprising administering to a subject with an MYD88 mutated disease. An MYD88 mutated disease can include, but is not limited to a proliferative disease (e.g., an IgM gammopathy (e.g., an 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, stomach cancer, lymphoma (e.g., B-cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgM secreting lymphoplasmacytic lymphoma (i.e., Waldenström'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 myelomas (e.g., IgM multiple myeloma)), and leukemia (e.g., chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia, myelogenous leukemia (e.g., chronic myelogenous leukemia, acute myelogenous leukemia (e.g., mast cell leukemia), myeloproliferative diseases (e.g., myelodysplastic syndrome))))).

[0242] In certain embodiments, the method described herein is superior (e.g., showing improved safety and/or therapeutic effects; decreased adverse effects) or comparable to existing therapy (e.g., chemotherapy).

[0243] In certain embodiments, the biological sample or cell (e.g., the biological sample or cell being contacted with a compound or pharmaceutical composition described herein) is in vitro. In certain embodiments, the cell is in vivo. In certain embodiments, the biological sample or cell is ex vivo.

[0244] In certain embodiments, the cell is a malignant cell (e.g., 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 a malignant white blood cell.

[0245] In certain embodiments, the cell is an adenocarcinoma cell, blastoma cell, carcinoma cell, or sarcoma cell. In certain embodiments, the cell is a pre-malignant cell (e.g., pre-cancerous cell).

[0246] In certain embodiments, the method described herein further comprises administering to the subject in need thereof an additional therapy. In certain embodiments, the additional therapy is an additional pharmaceutical agent described herein. In certain embodiments, the additional therapy is a cytotoxic chemotherapy (e.g., gemcitabine, cytarabine, daunorubicin, doxorubicin, vincristine, 1-asparaginase, cyclophosphamide, or etoposide). In certain embodiments, the additional therapy is an epigenetic modifier (e.g., azacitidine or romidepsin). In certain embodiments, the additional therapy is a glucocorticoid.

[0247] In certain embodiments, the additional therapy is an immunotherapy (e.g., an immunotherapeutic monoclonal antibody). In some embodiments, the additional pharmaceutical agent is etoposide, obatoclax, or navitoclax, and optionally the disease is breast cancer, e.g., 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 some embodiments, the additional pharmaceutical agent is etoposide, JIB04, or cisplatin. In some embodiments, the additional pharmaceutical agent is JQ1 or NVP2, and optionally the disease is leukemia, e.g., acute myelogenous leukemia, myeloblastic leukemia, promyelocytic leukemia, myelomonocytic leukemia, monocytic leukemia, monoblastic leukemia, or megakaryoblastic leukemia.

[0248] In yet another aspect, the present disclosure provides compounds and pharmaceutical compositions described herein for use in the treatment of a disease (e.g., a proliferative disease, such as an IgM gammopathy, mastocytosis, or cancer) in a subject in need thereof.

[0249] In yet another aspect, the present disclosure provides compounds and pharmaceutical compositions described herein for use in the prevention of a disease (e.g., a proliferative disease, such as an IgM gammopathy, mastocytosis, or cancer) in a subject in need thereof.

[0250] In another aspect, the present disclosure provides compounds and pharmaceutical compositions described herein for use in inhibiting the activity of a kinase (e.g., SRC Family kinases (e.g., HCK, LYN, BLK, FRK), Tec family kinases (e.g., BTK)) in a subject in need thereof.

[0251] In another aspect, the present disclosure provides compounds and pharmaceutical compositions described herein for use in inhibiting the activity of a kinase in a biological sample (e.g., an in vivo or ex vivo biological sample).

[0252] In another aspect, the present disclosure provides compounds and pharmaceutical compositions described herein for use in inhibiting the activity of a kinase in a cell (e.g., an in vivo or ex vivo cell).

[0253] In another aspect, the present disclosure provides uses of compounds and pharmaceutical compositions described herein in the manufacture of a medicament for treating a disease in a subject in need thereof.

[0254] In another aspect, the present disclosure provides uses of compounds and pharmaceutical compositions described herein in the manufacture of a medicament for preventing a disease in a subject in need thereof.

EXAMPLES

[0255] In order that the disclosure described herein may be more fully understood, the following examples are set forth. The synthetic and biological examples described in this application are offered to illustrate the compounds, pharmaceutical compositions, and methods provided herein and are not to be construed in any way as limiting their scope.

Example 1. Dose Response Curves for HCK Inhibitors

[0256] In vitro cellular efficacies (drug dose-response) were measured by CellTiter-Glo™ cell viability assay in MYD88 mutated BCWM.1, MWCL-1 WM cells; TMD8, HBL-1 ABC DLBCL cells; MYD88 wild type OCI-Ly7, OCI-Ly19 GCB DLBCL cells; Ramos Burkitt's lymphoma cells; and multiple myeloma cells. Cells were treated with either Compound I-4 (FIG. 2) or Compound I-1 (FIG. 3) with serially diluted drugs for 72 hrs (FIG. 2).

Example 2. Dose Response Curves for HCK Inhibitors

[0257] In vitro cellular efficacies (drug dose-response) of ibrutinib or Compound I-4 were measured by CellTiter-Glo™ cell viability assay in vector only, BTK WT or BTK C481S expressing lentiviral vector transduced TMD8 ABC-DLBCL or BCWM.1 WM cells. Cells were treated with serially diluted ibrutinib or SB1-G-114 for 72 hrs. The results show that Compound I-4 overcomes BTKC481S mediated ibrutinib resistance in MYD88 mutated WM and ABC DLBCL cells (FIG. 4).

Example 3. ED.SUB.50 .Values

[0258] ED.sub.50 values of the different compounds were measured by CellTiter-Glo™ cell viability assay in MYD88 mutated BCWM.1, MWCL-1 WM cells; TMD8, HBL-1 ABC DLBCL cells; MYD88 wild type OCI-Ly7, OCI-Ly19 GCB DLBCL cells; Ramos Burkitt's lymphoma cells; and RPMI-8226 multiple myeloma cells. Cells were treated with serially diluted drugs for 72 hrs

TABLE-US-00002 TABLE 1 ED.sub.50 values of compounds # BCWM.1 MWCL-1 TMD8 HBL-1 OCI-Ly7 OCI-Ly19 Ramos RPMI-8226 cis I-1 7.50E−07 9.86E−07 8.26E−07 5.88E−06 2.31E−06 4.46E−06 4.44E−06 2.36E−05 trans I-1 9.99E−09 1.00E−08 3.52E−08 3.59E−07 4.33E−08 8.59E−06 1.15E−07 8.02E−05 cis I-3 6.51E−07 8.43E−07 6.24E−07 1.01E−06 7.94E−07 1.17E−06 8.10E−07 1.02E−06 trans I-3 9.73E−07 1.33E−06 6.84E−07 1.58E−06 7.11E−07 8.38E−07 8.81E−07 2.12E−06 cis I-4 1.00E−06 8.10E−07 7.57E−07 1.60E−06 1.36E−06 1.70E−06 1.31E−06 3.05E−06 trans I-4 1.09E−07 2.28E−08 9.54E08  5.28E−07 8.28E−07 1.96E−06 8.94E−07 3.11E−06 I-5 4.39E−06 2.30E−06 2.99E−06 1.27E−05 7.48E−06 1.61E−05 2.84E−05 4.27E−05 trans I-6 4.24E−07 1.40E−07 7.19E−07 2.23E−05 7.54E−07 2.33E−05 8.56E−05 1.49E−02 cis I-6 2.47E−06 1.21E−06 1.75E−06 5.89E−06 5.24E−06 9.14E−06 7.32E−06 1.76E−05 trans I-7 N 8.85E−05 2.37E−07 1.83E−06 1.73E−06 1.16E−06 8.73E−06 1.40E−05 cis I-7 4.18E−07 2.95E−06 2.71E−07 6.13E−06 3.97E−06 4.85E−06 7.20E−06 3.40E−05 trans I-8 7.14E−06 4.91E−06 4.80E−06 6.81E−06 3.39E−06 6.24E−06 6.87E−05 2.14E−05 cis I-8 1.57E−05 1.41E−05 N 1.68E−05 2.09E−05 1.44E−05 4.33E−05 2.40E−05 trans I-9 N N 3.32E−06 7.63E−06 2.79E−04 N N N cis I-9 5.38E−06 N 6.44E−06 7.98E−06 5.99E−06 5.37E−06 1.61E−05 1.20E−04 trans I-10 8.04E−07 3.50E−06 1.56E−05 8.61E−06 2.15E−06 N 4.21E−06 3.02E−04 cis I-10 1.06E−06 1.02E−06 8.06E−07 1.85E−06 1.10E−06 1.53E−06 6.58E−07 1.38E−06

Example 4. Synthesis of Cis and Trans I-4

[0259] cis 1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (cis I-4) and trans 1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (trans I-4) were synthesized according to the following scheme.

##STR00085## ##STR00086##

4-chloro-3-iodo-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[4,3-c]pyridine (1)

[0260] A mixture of 4-chloro-3-iodo-1H-pyrazolo[4,3-c]pyridine (3 g, 10.7 mmol), 1,4-dioxaspiro[4.5]decan-8-yl 4-methylbenzenesulfonate (5 g, 16.1 mmol), and Cs.sub.2CO.sub.3 (7 g, 21.4 mmol) in DMF (100 mL) was stirred at 90° C. for 16 h. After the reaction was complete, the mixture was concentrated and purified by silica gel (DCM/MeOH=10/1) to obtain compound 1 (white solid, 700 mg, 16% yield). LCMS: 420 (M+H).sup.+.

N-(2,4-dimethoxybenzyl)-3-iodo-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine (2)

[0261] A mixture of 4-chloro-3-iodo-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[4,3-c]pyridine 1 (400 mg, 0.95 mmol), (2,4-dimethoxyphenyl)methanamine (319 mg, 1.91 mmol), and Cs.sub.2CO.sub.3 (620 mg, 1.91 mmol) was stirred at 125° C. for 16 h. After the reaction was complete, the mixture was concentrated and purified by silica gel (DCM/MeOH=10/1) to obtain compound 2 (yellow solid, 500 mg, 95% yield). LCMS: 551 (M+H).sup.+.

N-(2,4-dimethoxybenzyl)-3-(4-phenoxyphenyl)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine (3)

[0262] A mixture of N-(2,4-dimethoxybenzyl)-3-iodo-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine 2 (250 mg, 0.45 mmol), K.sub.2CO.sub.3 (120 mg, 0.9 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (150 mg, 0.45 mmol), and 4,4,5,5-tetramethyl-2-(4-phenoxyphenyl)-1,3,2-dioxaborolane (130 mg, 0.45 mmol) in THF (5 mL) and H.sub.2O (0.1 mL) was stirred at 60° C. for 16 h. After the reaction was complete the mixture was concentrated and purified by prep-HPLC (C18 column, CH.sub.3CN/H.sub.2O, containing 0.05% NH.sub.4HCO.sub.3) to get title compound 3 (white solid, 170 mg, 63%). LCMS: 593 (M+H).sup.+.

4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[4,3-c]pyridin-1-yl)cyclohexanone (4)

[0263] The mixture of N-(2,4-dimethoxybenzyl)-3-(4-phenoxyphenyl)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine 3 (238 mg, 0.4 mmol) in TFA (5 mL), and DCM (2 mL) was stirred at 45° C. for 16 h. After the reaction was complete, the mixture was concentrated and purified by prep-HPLC (C18 column, CH.sub.3CN/H.sub.2O, containing 0.05% NH.sub.4HCO.sub.3) to get compound 4 (white solid, 150 mg, 94%). LCMS: 399 (M+H).sup.+.

cis 1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (cis I-4) and trans 1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (trans I-4)

[0264] A mixture of 4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[4,3-c]pyridin-1-yl) cyclohexanone (150 mg, 0.25 mmol), 1-methylpiperazine (100 mg, 1.00 mmol), and NaBH(OAc).sub.3 (212 mg, 1.00 mmol) in THF (5 mL) was stirred at rt for 16 h. After the reaction was complete the mixture was concentrated and purified by prep-HPLC (C18 column, CH.sub.3CN/H.sub.2O, containing 0.05% NH.sub.4HCO.sub.3) to get title compound cis I-4 (white solid, 4 mg, 2.2%) and trans I-4 (white solid, 3 mg, 2.1%). HPLC: 100% (@254 nm);

[0265] cis I-4: LCMS: 483 (M+H).sup.+; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.83 (d, J=6.2 Hz, 1H), 7.75-7.55 (m, 2H), 7.37 (tt, J=39.2, 19.7 Hz, 2H), 7.21-6.84 (m, 5H), 6.82 (d, J=6.2 Hz, 1H), 4.97 (s, 2H), 4.71-4.20 (m, 1H), 3.00-2.34 (m, 9H), 2.33-2.12 (m, 7H), 1.82-1.68 (m, 2H), 1.60 (dd, J=14.0, 11.1 Hz, 2H).

[0266] trans I-4: LCMS: 483 (M+H).sup.+; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.83 (d, J=6.2 Hz, 1H), 7.70-7.56 (m, 2H), 7.46-7.32 (m, 2H), 7.20-6.97 (m, 5H), 6.72 (d, J=6.3 Hz, 1H), 5.17 (d, J=103.3 Hz, 2H), 4.58-4.07 (m, 1H), 2.67 (s, 4H), 2.59-2.34 (m, 5H), 2.30 (s, 3H), 2.23-1.92 (m, 6H), 1.51 (dd, J=11.7, 4.5 Hz, 2H).

Example 5. Synthesis of Cis and Trans I-3

[0267] Synthesis of cis 1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrrolo[3,2-c]pyridin-4-amine (cis I-3) and trans 1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrrolo[3,2-c]pyridin-4-amine (trans I-3) were synthesized following the same procedure as cis and trans I-4, but starting with 4-chloro-3-iodo-1H-pyrrolo[3,2-c]pyridine instead.

[0268] cis I-3: LCMS (m/z): 482 [M+H].sup.+; .sup.1H NMR (400 MHz, DMSO) δ 7.73 (d, J=6.4 Hz, 2H), 7.46 (t, J=8.8 Hz, 2H), 7.38 (t, J=8.0 Hz, 2H), 7.15 (t, J=7.6 Hz, 1H), 7.07-7.10 (m, 5H), 6.83 (d, J=6.0 Hz, 1H), 5.09 (br, 2H), 4.26 (m, 1H), 2.48-2.51 (m, 7H), 2.31 (m, 4H), 2.13-2.25 (m, 5H), 1.55-1.62 (m, 3H), 1.25 (m, 1H).

[0269] trans I-3: LCMS (m/z): 482 [M+H]+; .sup.1H NMR (400 MHz, DMSO) δ 7.72 (d, J=6.4 Hz, 2H), 7.43 (t, J=8.4 Hz, 2H), 7.38 (t, J=8.0 Hz, 2H), 7.15 (t, J=7.6 Hz, 1H), 7.01-7.08 (m, 5H), 6.75 (d, J=6.4 Hz, 1H), 5.27 (br, 2H), 4.14 (m, 1H), 2.40-2.67 (m, 8H), 2.31 (s, 3H), 2.12-2.25 (m, 4H), 1.53-1.59 (m, 3H), 1.31 (m, 2H).

Example 6. Synthesis of Cis and Trans I-5

[0270] 6-amino-9-(4-(4-methylpiperazin-1-yl)cyclohexyl)-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one (I-5) was synthesized according to the following scheme.

##STR00087## ##STR00088##

N,N-dibenzyl-6-chloro-5-nitropyrimidin-4-amine (1a)

[0271] To a mixture of 4,6-dichloro-5-nitropyrimidine (10 g, 51.8 mmol) and DIPEA (13.4 g, 103.6 mmol) in DCM (40 mL) was added dropwise a solution of dibenzylamine (10.2 g, 51.8 mmol) in DCM (20 mL) at 0° C., and the mixture was stirred at rt. After 16 h, the reaction was concentrated and purified by column chromatography (silica gel, EA/PE=1/6) to afford the desired product 1a as yellow solid (15.0 g, yield 90%). LCMS (m/z): 355.0 [M+H].sup.+.

N.SUP.4.,N.SUP.4.-dibenzyl-5-nitro-N.SUP.6.-(1,4-dioxaspiro[4.5]decan-8-yl)pyrimidine-4,6-diamine (2a)

[0272] To the mixture of N,N-dibenzyl-6-chloro-5-nitropyrimidin-4-amine 1 (15 g, 42.3 mmol) and DIPEA (10.2 g, 84.6 mmol) in DCM (40 mL) was added dropwise the solution of 1,4-dioxaspiro[4.5]decan-8-amine (6.6 g, 42.3 mmol) in DCM (20 mL), the mixture was stirred at rt for 16 h, concentrated and purified by column chromatography (silica gel, EA/PE=1/2) to afford desired product 2a as yellow solid (13.0 g, yield 65%). LCMS (m/z): 476.1 [M+H]+.

N.SUP.4.,N.SUP.4.-dibenzyl-N.SUP.6.-(1,4-dioxaspiro[4.5]decan-8-yl)pyrimidine-4,5,6-triamine (3a)

[0273] A mixture of N.sup.4,N.sup.4-dibenzyl-5-nitro-N.sup.6-(1,4-dioxaspiro[4.5]decan-8-yl)pyrimidine-4,6-diamine 2a (10.0 g, 21.0 mmol), Fe (5.8 g, 105.0 mmol) and NH.sub.4Cl (5.9 g, 105.0 mmol) in EtOH (50 mL) was stirred at 80° C. After 3 h, the mixture was filtered, and the filtrate was concentrated to leave crude product 3a as yellow solid (8.0 g crude, yield 86%). LCMS (m/z): 446.1 [M+H].sup.+.

6-(dibenzylamino)-9-(1,4-dioxaspiro[4.5]decan-8-yl)-7,9-dihydro-8H-purin-8-one (4a)

[0274] To a mixture of N.sup.4,N.sup.4-dibenzyl-N6-(1,4-dioxaspiro[4.5]decan-8-yl)pyrimidine-4,5,6-triamine 3a (6.0 g, 13.4 mmol) and DIPEA (5.0 mL, 26.8 mmol) in DCM (60 mL) was added Triphosgene (bis(trichloromethyl) carbonate (BTC) (1.6 g, 5.4 mmol) at 0° C. The mixture was then stirred at 40° C. for 3 h, concentrated, and purified by column chromatography (silica gel, MeOH/DCM=1/20) to afford product 4a as yellow solid (3.0 g, yield 47%). LCMS (m/z): 472.1 [M+H].sup.+.

6-amino-9-(1,4-dioxaspiro[4.5]decan-8-yl)-7,9-dihydro-8H-purin-8-one (5a)

[0275] A mixture of 6-(dibenzylamino)-9-(1,4-dioxaspiro[4.5]decan-8-yl)-7H-purin-8(9H)-one 4a (2.5 g, 5.3 mmol) and Pd/C (10%, 0.3 g) in EtOH (50 mL) was stirred at 60° C. under hydrogen (1 atm) for 16 h. The mixture was then filtered through celite, and the filtrate was concentrated and purified by column chromatography (silica gel, EA/PE=3/1) to afford product 5a as yellow solid (1.1 g, yield 73%). LCMS (m/z): 292.0 [M+H].sup.+.

6-amino-7-(4-phenoxyphenyl)-9-(1,4-dioxaspiro[4.5]decan-8-yl)-7,9-dihydro-8H-purin-8-one (6a)

[0276] A mixture of 6-amino-9-(1,4-dioxaspiro[4.5]decan-8-yl)-7H-purin-8(9H)-one 5 (1.1 g, 3.8 mmol), (4-phenoxyphenyl)boronic acid (1.6 g, 7.6 mmol), Cu(OAc).sub.2 (1.4 g, 7.6 mmol) and DIPEA (1.3 mL, 7.6 mmol) in DCM (30 mL) was stirred at 40° C. for 16 h. The mixture was then filtered, and the filtrate was concentrated and purified by column chromatography (silica gel, MeOH/DCM=1/20) to obtain product 6a as yellow solid (0.8 g, yield 47%). LCMS (m/z): 460.1 [M+H].sup.+.

6-amino-9-(4-oxocyclohexyl)-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one (7a)

[0277] A mixture of 6-amino-7-(4-phenoxyphenyl)-9-(1,4-dioxaspiro[4.5]decan-8-yl)-7H-purin-8(9H)-one 6a (0.7 g, 1.5 mmol) and concentrated HCl solution (8.0 mL) in acetone (12 mL) was stirred at rt for 3 h. The mixture was then concentrated under reduced pressure, and the residue was diluted with water (10 mL). The solution was adjusted to pH 8 with Na.sub.2CO.sub.3 and extracted with ethyl acetate (150 mL×2), and the combined organic was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to leave crude product 7a as yellow solid (0.8 g).

6-amino-9-(4-(4-methylpiperazin-1-yl)cyclohexyl)-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one (I-5)

[0278] A mixture of 6-amino-9-(4-oxocyclohexyl)-7-(4-phenoxyphenyl)-7H-purin-8(9H)-one 7 (0.5 g, 1.2 mmol), 1-methylpiperazine (2.4 g, 24 mmol) and CH.sub.3COOH (0.1 mL) in DCE (10 mL) was stirred at rt for 3 h. After this time, NaBH(OAc).sub.3 (1.0 g, 4.8 mmol) was added, and the mixture was stirred at rt overnight and concentrated in vacuum. The residue was then diluted with ethyl acetate (150 mL), washed with brine (50 mL×2), dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated and purified by preparative HPLC (C18 column, CH.sub.3CN/H.sub.2O, containing 0.05% NH.sub.4HCO.sub.3) to afford I-5 as white solid (64.1 mg, yield 10%). LCMS (m/z): 500.1 [M+H].sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz): δ (ppm) 8.12 (s, 1H), 7.45-7.41 (m, 4H), 7.19 (t, J=7.6 Hz, 1H), 7.13 (t, J=6.4 Hz, 4H), 5.73 (s, 2H), 4.32 (t, J=12.0 Hz, 1H), 2.67-2.57 (m, 3H), 2.35 (m, 6H), 2.15 (s, 3H), 2.12-2.06 (m, 4H), 1.47-1.41 (m, 4H).

Example 7. Synthesis of Cis and Trans I-6

[0279] The synthesis of trans 4-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one (trans I-6) and cis 4-amino−1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one (cis I-6) was carried out according to the following scheme.

##STR00089## ##STR00090##

2-chloro-3-nitro-N-(1,4-dioxaspiro[4.5]decan-8-yl)pyridin-4-amine (1b

[0280] To a mixture of 2,4-dichloro-3-nitropyridine (8.5 g, 44.2 mmol) and DIPEA (11.4 g, 88.4 mmol) in EtOH (40 mL) was added dropwise a solution of 1,4-dioxaspiro[4.5]decan-8-amine (6.9 g, 44.2 mmol) in EtOH (10 mL) at 0° C. The mixture was then stirred at rt for 16 h, concentrated and purified by column chromatography (silica-gel, EA/PE=1/2) to afford product 1b as yellow solid (5.1 g, yield 37%). LCMS (m/z): 314.0 [M+H].sup.+.

N.SUP.2.,N.SUP.2.-dibenzyl-3-nitro-N.SUP.4.-(1,4-dioxaspiro[4.5]decan-8-yl)pyridine-2,4-diamine (2b)

[0281] A mixture of 2-chloro-3-nitro-N-(1,4-dioxaspiro[4.5]decan-8-yl)pyridin-4-amine 1b (5.1 g, 16.3 mmol), dibenzylamine (3.2 g, 16.3 mmol), and K.sub.2CO.sub.3 (4.4 g, 32.6 mmol) in DMF (50 mL) was stirred at 120° C. for 16 h. The mixture was then cooled down to rt, diluted with water (200 mL) and extracted with ethyl acetate (150 mL×2). The combined organic extract was washed with brine (50 mL×4), dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated, and purified by column chromatography (silica-gel, EA/PE=1/2) to afford product 2b as yellow solid (6.0 g, yield 80%). LCMS (m/z): 475.1 [M+H].sup.+.

N.SUP.2.,N.SUP.2.-dibenzyl-N.SUP.4.-(1,4-dioxaspiro[4.5]decan-8-yl)pyridine-2,3,4-triamine (3b)

[0282] A mixture of N.sup.2,N.sup.2-dibenzyl-3-nitro-N4-(1,4-dioxaspiro[4.5]decan-8-yl)pyridine-2,4-diamine 2b (5.0 g, 10.5 mmol), Fe (2.9 g, 52.0 mmol), and NH.sub.4Cl (3.0 g, 52.0 mmol) in EtOH (30 mL) and H.sub.2O (5 mL) was stirred at 80° C. for 3 h. The mixture was then filtered, and the filtrate was concentrated to leave crude product 3b as yellow solid (4.2 g crude, yield 91%). LCMS (m/z): 445.1 [M+H].sup.+.

4-(dibenzylamino)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one (4b)

[0283] To a mixture of 3b (4.2 g, 9.5 mmol) and DIPEA (4.0 mL, 19 mmol) in DCM (60 mL) was added BTC (1.2 g, 3.7 mmol) at 0° C. The mixture was then stirred at 40° C. for 3 h, concentrated, and purified by column chromatography (silica-gel, MeOH/DCM=1/20) to afford product 4b as yellow solid (4.0 g, yield 90%). LCMS (m/z): 471.1 [M+H].sup.+.

4-amino-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one (5b)

[0284] A mixture of 4b (2.0 g, 4.2 mmol) and Pd/C (10%, 0.2 g) in EtOH (30 mL) was stirred at 60° C. for 16 h. The mixture was then filtered through celite, concentrated, and purified by column chromatography (silica-gel, MeOH/DCM=1/10) to afford product 5b as a yellow solid (1.0 g, yield 83%). LCMS (m/z): 291.0 [M+H].sup.+.

4-amino-3-(4-phenoxyphenyl)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one (6b)

[0285] A mixture of 5b (1.0 g, 3.4 mmol), (4-phenoxyphenyl)boronic acid (1.4 g, 6.8 mmol), Cu(OAc).sub.2 (1.2 g, 6.8 mmol) and DIPEA (1.5 mL, 6.8 mmol) in DCM (30 mL) was stirred at 40° C. for 48 h. The mixture was then filtered, concentrated, and purified by column chromatography (silica-gel, MeOH/DCM=1/10) and preparative HPLC (C18 column, CH.sub.3CN/H.sub.2O, containing 0.05% NH.sub.4HCO.sub.3) to afford product 6b as yellow solid (0.5 g, yield 31%). LCMS (m/z): 459.1 [M+H].sup.+.

4-amino-1-(4-oxocyclohexyl)-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one (7b)

[0286] A mixture of 6b (0.5 g, 1.09 mmol) and TFA (8.0 mL) in DCM (8.0 mL) was stirred at 50° C. for 3 h. The mixture was then concentrated in vacuo, and the residue was diluted with water (100 mL), and the pH adjusted to 8 with Na.sub.2CO.sub.3. This mixture was then extracted with ethyl acetate (50 mL×2), and the combined organic was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to leave crude product 7b as yellow solid (0.31 g, yield 68%).

trans 4-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one (trans I-6) and cis 4-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one (cis I-6)

[0287] A mixture of 7b (0.3 g, 0.72 mmol), 1-methylpiperazine (1.4 g, 14.4 mmol) and CH.sub.3COOH (0.1 mL) in DCE (10 mL) was stirred at rt for 3 h. After this time, NaBH(OAc).sub.3 (0.59 g, 2.8 mmol) was added, and the resulting mixture was stirred at rt overnight. The mixture was then concentrated in vacuo, and the residue was diluted with ethyl acetate (50 mL), washed with brine (50 mL×2), dried over anhydrous Na.sub.2SO.sub.4, concentrated, and purified by preparative HPLC (C18 column, CH.sub.3CN/H.sub.2O, containing 0.05% NH.sub.4HCO.sub.3) to afford product trans I-6 as white solid (41.8 mg) and cis I-6 as white solid (88.6 mg).

[0288] trans I-6: LCMS (m/z): 499.1 [M+H].sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz): δ (ppm) 7.73 (d, J=5.6 Hz, 1H), 7.47-7.41 (m, 4H), 7.21 (t, J=7.6 Hz, 1H), 7.15-7.10 (m, 4H), 6.88 (d, J=5.6 Hz, 1H), 4.79 (s, 2H), 4.15 (t, J=12.4 Hz, 1H), 2.64 (m, 2H), 2.46-2.10 (m, 11H), 1.91-1.79 (m, 4H), 1.46-1.37 (m, 2H).

[0289] cis I-6: LCMS (m/z): 499.1 [M+H].sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz): δ (ppm) 7.76 (d, J=5.2 Hz, 1H), 7.47-7.42 (m, 4H), 7.21 (t, J=6.8 Hz, 1H), 7.13 (t, J=9.6 Hz, 4H), 7.01 (d, J=5.6 Hz, 1H), 4.82 (s, 2H), 4.36 (m, 1H), 2.44-2.07 (m, 15H), 1.53-1.47 (m, 4H).

Example 8. Synthesis of cis and trans I-1

[0290] cis-5-Amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (cis I-1) and trans 5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (trans I-1) were synthesized according to the below scheme.

##STR00091##

5-amino-3-(4-phenoxyphenyl)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazole-4-carbonitrile (1c)

[0291] A mixture of 2-(methoxy(4-phenoxyphenyl)methylene)malononitrile (0.94 g, 3.4 mmol) and 1,4-dioxaspiro[4.5]decan-8-ylhydrazine (1.7 g, 10.2 mmol) in EtOH (30 mL) was stirred at 65° C. for 16 h. The mixture was then concentrated and purified by column chromatography (silica gel, EA/PE=2/1) to afford product 1c as a yellow solid (1.0 g, yield 71%). LCMS (m/z): 417.1 [M+H].sup.+.

5-amino-1-(4-oxocyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile (2c)

[0292] A mixture of 1c (1.0 g, 2.4 mmol) and TFA (10 mL) in DCM (10 mL) was stirred at 50° C. for 16 h. The mixture was then concentrated to remove the solvents, and the residue was diluted with water (100 mL), adjusted to pH 8 with NaHCO.sub.3 and extracted with ethyl acetate (150 mL×2). The combined organic extract was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to leave crude product 2c as yellow oil (1.2 g). LCMS (m/z): 373.1 [M+H].sup.+.

Cis 5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile (cis 3c) and trans 5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile (trans 3c)

[0293] A mixture of 2c (1.2 g, 2.4 mmol), 1-methylpiperazine (1.5 mL, 9.6 mmol), and CH.sub.3COOH (0.3 mL) in THF (20 mL) was stirred at 70° C. for 1 h. After that time, the reaction mixture was cooled to rt and NaBH(OAc).sub.3 (2.0 g, 9.6 mmol) was then added in small batches, and the resulting mixture was stirred at rt overnight. The reaction mixture was then concentrated, and the residue was diluted with ethyl acetate (150 mL), washed with brine (50 mL×2), dried over anhydrous Na.sub.2SO.sub.4, concentrated, and purified by preparative TLC to afford cis 3c as yellow oil (25 mg) and trans 3c as yellow oil (200 mg). LCMS (m/z): 457.3 [M+H].sup.+.

cis 5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (cis I-1)

[0294] A mixture of cis 3c (25 mg, 0.05 mmol) and NaOH (9 mg, 0.2 mmol) in EtOH (2 mL) and H.sub.2O (0.5 mL) was heated in a microwave reactor at 150° C. for 3 h. The mixture was then concentrated and purified by preparative HPLC (C18 column, CH.sub.3CN/H.sub.2O, containing 0.05% NH.sub.4HCO.sub.3) to obtain cis I-1 as a white solid (5.5 mg). LCMS (m/z): 475.1 [M+H].sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz): δ (ppm) 7.47 (d, J=8.8 Hz, 2H), 7.42 (t, J=8.0 Hz, 2H), 7.17 (t, J=7.2 Hz, 1H), 7.07 (t, J=9.2 Hz, 4H), 6.30 (s, 2H), 4.17-4.12 (m, 1H), 2.33-2.22 (m, 7H), 2.12 (s, 4H), 2.07-1.99 (m, 4H), 1.56-1.43 (m, 4H).

trans 5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (trans I-1)

[0295] A mixture of trans 5-amino-1-(−4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile trans 3c (100 mg, 0.22 mmol) and NaOH (36 mg, 0.88 mmol) in EtOH (4 mL) and H.sub.2O (2 mL) was heated in a microwave reactor at 150° C. for 3 h. The mixture was then concentrated and purified by preparative HPLC (C18 column, CH.sub.3CN/H.sub.2O, containing 0.05% NH.sub.4HCO.sub.3) to obtain trans I-1 as a white solid (23.6 mg). LCMS (m/z): 475.1 [M+H].sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz): δ (ppm) 7.47 (d, J=8.8 Hz, 2H), 7.41 (t, J=7.6 Hz, 2H), 7.16 (t, J=7.2 Hz, 1H), 7.08-7.03 (m, 4H), 6.30 (s, 2H), 4.05 (t, J=10.8 Hz, 1H), 2.42-2.28 (m, 8H), 2.12 (s, 3H), 1.88-1.74 (m, 6H), 1.40-1.32 (m, 2H).

Example 9. Synthesis of Cis and Trans I-9

[0296] The synthesis of trans 5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one (trans I-9) and cis 5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one (cis I-9) was carried out according to the following scheme.

##STR00092## ##STR00093##

6-chloro-5-(((4-phenoxyphenyl)amino)methyl)pyrimidin-4-amine (1d)

[0297] A mixture of 4-amino-6-chloropyrimidine-5-carbaldehyde (3.0 g, 19.1 mmol), 4-phenoxyaniline (3.8 g, 21.0 mmol), and CH.sub.3COOH (2.0 mL) in DCE (20 mL) was stirred at rt for 6 h. After this time, NaBH(OAc).sub.3 (20 g, 95.5 mmol) was added in small batches, and the resulting mixture was stirred at rt overnight. The mixture was then concentrated, and the residue was diluted with ethyl acetate (150 mL), washed with brine (50 mL×2), dried over anhydrous Na.sub.2SO.sub.4, concentrated, and purified by column chromatography (silica gel, MeOH/DCM=1/20) to afford desired product 1d as yellow solid (1.2 g, yield 19%). LCMS (m/z): 326.9 [M+H].sup.+.

5-chloro-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one (2d)

[0298] To a mixture of 1d (1.2 g, 3.68 mmol) and DIPEA (3.0 mL, 7.36 mmol) in DCM (30 mL) was added dropwise the solution of bis(trichloromethyl) carbonate (0.65 g, 2.2 mmol) in DCM (15 mL) at 0° C. The mixture was then stirred at rt for 16 h. After this time, the mixture was concentrated and purified by column chromatography (silica gel, EA/PE=7/1) to afford 2d as a yellow solid (0.77 g, yield 59%). LCMS (m/z): 352.9 [M+H].sup.+.

5-(bis(4-methoxybenzyl)amino)-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one (3d)

[0299] A mixture of 5-chloro-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one (0.7 g, 2.0 mmol), bis(4-methoxybenzyl)amine (1.02 g, 4.0 mmol) and DIPEA (0.5 g, 4.0 mmol) in 2-BuOH (20 mL) was stirred at 110° C. for 16 h. The mixture was then concentrated, and the residue was purified by column chromatography (silica gel, MeOH/DCM=1/20) to afford desired product 3d as yellow solid (0.58 g, yield 52%). LCMS (m/z): 574.1 [M+H].sup.+.

5-(bis(4-methoxybenzyl)amino)-3-(4-phenoxyphenyl)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one (4d)

[0300] A mixture of 3d (0.5 g, 0.87 mmol), 1,4-dioxaspiro[4.5]decan-8-yl 4-methylbenzenesulfonate 7 (0.54 g, 1.74 mmol) and Cs.sub.2CO.sub.3 (0.56 g, 1.74 mmol) in DMF (8.0 mL) was stirred at 100° C. for 48 h. The mixture was then diluted with water (100 mL) and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with brine (50 mL×2), dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated and purified by column chromatography (silica gel, EA/PE=1/1) to afford 4d as white solid (0.34 g, yield 50%). LCMS (m/z): 714.0 [M+H].sup.+.

5-amino-1-(4-oxocyclohexyl)-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one (5d)

[0301] A mixture of 4d (0.34 g, 0.47 mmol) and TFA (6.0 mL) in DCM (6.0 mL) was stirred at 65° C. for 16 h. After this time, the mixture was concentrated, and the residue was diluted with water (20 mL), adjusted to pH 8 with Na.sub.2CO.sub.3, and extracted with ethyl acetate (50 mL×2). The combined organic was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to leave crude product 5d as yellow oil (0.2 g).

trans 5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one (trans I-9) and cis 5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one (cis I-9)

[0302] A mixture of 5d (0.15 g, 0.35 mmol), 1-methylpiperazine (0.7 g, 7.0 mmol) and CH.sub.3COOH (0.1 mL) in DCE (10 mL) was stirred at rt for 3 h, and then NaBH(OAc).sub.3 (0.37 g, 1.75 mmol) was added. The mixture was then stirred at rt overnight and concentrated in vacuum. The residue was then diluted with ethyl acetate (50 mL), washed with brine (50 mL×2), dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated, and purified by preparative HPLC (C18 column, CH.sub.3CN/H.sub.2O, containing 0.05% NH.sub.4HCO.sub.3) to afford trans I-9 as white solid (0.5 mg) and cis I-9 as a white solid (37.7 mg).

[0303] trans I-9: LCMS (m/z): 514.0 [M+H].sup.+; .sup.1H NMR (CDCl.sub.3, 400 MHz): δ (ppm) 8.32 (s, 1H), 7.36-7.27 (m, 4H), 7.14-7.01 (m, 5H), 4.60 (s, 2H), 4.51 (s, 2H), 2.65-2.58 (m, 10H), 2.30 (s, 3H), 2.00-1.96 (m, 2H), 1.82-1.79 (m, 2H), 1.26-1.22 (m, 4H).

[0304] cis I-9: LCMS (m/z): 514.0 [M+H].sup.+; .sup.1H NMR (CDCl.sub.3, 400 MHz): δ (ppm) 8.32 (s, 1H), 7.34 (t, J=7.6 Hz, 2H), 7.28-7.27 (m, 2H), 7.12 (t, J=7.6 Hz, 1H), 7.03-7.01 (m, 4H), 4.81-4.70 (m, 2H), 4.82 (s, 2H), 2.81-2.46 (m, 9H), 2.26 (s, 3H), 2.18-2.06 (m, 5H), 1.50-1.44 (m, 4H).

Example 10. Synthesis of Cis and Trans I-10

[0305] The synthesis of trans 4-amino-8-(4-(4-methylpiperazin-1-yl)cyclohexyl)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one (trans I-10) and cis 4-amino-8-((1r,4r)-4-(4-methylpiperazin-1-yl)cyclohexyl)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one (cis I-10) was carried out according to the below scheme.

##STR00094## ##STR00095##

4-amino-6-chloro-2-(methylthio)pyrimidine-5-carbaldehyde (1e)

[0306] A mixture of 4,6-dichloro-2-(methylthio)pyrimidine-5-carbaldehyde (1.5 g, 6.7 mmol), NH.sub.4OH (6.0 mL) and DCM (20 mL) was stirred at rt for 3 h. The mixture was then washed with brine (50 mL×2), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to afford desired product 1e as a white solid (1.3 g, yield 90%). LCMS (m/z): 203.9 [M+H].sup.+.

4-chloro-2-(methylthio)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one (2e)

[0307] A mixture of 1e (1.3 g, 6.4 mmol), methyl 2-(4-phenoxyphenyl)acetate (1.5 g, 6.4 mmol) and K.sub.2CO.sub.3 (1.8 g, 12.8 mmol) in NMP (12 mL) was stirred at 100° C. for 2 h. The mixture was then diluted with water (100 mL) and extracted with ethyl acetate (100 mL×2). The combined organic was washed with brine (50 mL×4), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to afford 2e as yellow solid (1.7 g, yield 68%). LCMS (m/z): 395.9 [M+H].sup.+.

4-amino-2-(methylthio)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one (3e)

[0308] The mixture of 4-chloro-2-(methylthio)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one 2 (0.8 g, 2.0 mmol) and NH.sub.4OH (4.0 mL) in 1,4-dioxane (10 mL) was stirred at 80° C. for 16 h, the mixture was concentrated in vacuum, the residue was slurried in a mixture of EA (10 mL) and PE (20 mL), filtered and dried under vacuum to afford desired product 3 as yellow solid (1.0 g). LCMS (m/z): 376.9 [M+H].sup.+. 4-amino-2-(methylthio)-6-(4-phenoxyphenyl)-8-(1,4-dioxaspiro[4.5]decan-8-yl)pyrido[2,3-d]pyrimidin-7(8H)-one (4e)

[0309] A mixture of 3e (1.0 g, 2.6 mmol), 1,4-dioxaspiro[4.5]decan-8-yl 4-methylbenzenesulfonate (1.65 g, 5.3 mmol) and Cs.sub.2CO.sub.3 (1.7 g, 5.3 mmol) in DMF (12 mL) was stirred at 100° C. for 16 h. The mixture was then diluted with water (100 mL) and extracted with ethyl acetate (100 mL×2). The combined organic extract was then washed with brine (50 mL×4), dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated and purified by column chromatography (silica gel, EA/PE=1/2) to afford desired product 4e as a yellow solid (0.2 g, yield 18%). LCMS (m/z): 516.9 [M+H].sup.+.

4-amino-2-(methylthio)-8-(4-oxocyclohexyl)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one (5e)

[0310] The mixture of 4e (0.2 g, 0.38 mmol) and TFA (5.0 mL) in DCM (5.0 mL) was stirred at rt for 2 h. The mixture was then concentrated, and the residue was diluted with water (20 mL), adjusted to pH 8 with Na.sub.2CO.sub.3, and extracted with ethyl acetate (50 mL×2). The combined organic extract was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to leave crude product 5e as a yellow solid (0.18 g). LCMS (m/z): 473.0 [M+H].sup.+.

4-amino-8-(4-(4-methylpiperazin-1-yl)cyclohexyl)-2-(methylthio)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one (6e)

[0311] The mixture of 5e (0.13 g, 0.28 mmol), 1-methylpiperazine (0.3 g, 2.8 mmol) and CH.sub.3COOH (0.1 mL) in DCE (10 mL) was stirred at rt for 3 h. NaBH(OAc).sub.3 (0.6 g, 2.8 mmol) was then added and the mixture was stirred at rt overnight. After this time, the mixture was concentrated, diluted with water (50 mL) and extracted with ethyl acetate (50 mL×2). The combined organic extract was washed with brine (50 mL×2), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to afford 6e as a yellow oil (200 mg). LCMS (m/z): 557.0 [M+H].sup.+.

trans 4-amino-8-(4-(4-methylpiperazin-1-yl)cyclohexyl)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one (trans I-10) and cis 4-amino-8-(4-(4-methylpiperazin-1-yl)cyclohexyl)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one (cis I-10)

[0312] A mixture of 6e (0.17 g, 0.31 mmol), Ni (0.5 g) and EtOH (10 mL) was stirred at 60° C. for 3 h. The mixture was then filtered through celite, and the filtrate was concentrated and purified by preparative HPLC (C18 column, CH.sub.3CN/H.sub.2O, containing 0.05% NH.sub.4HCO.sub.3) to afford desired product trans I-10 as a white solid (3.7 mg) and cis I-10 as white solid (2.0 mg).

[0313] trans I-10: LCMS (m/z): 511.0 [M+H].sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz): δ (ppm) 8.60 (s, 1H), 8.40 (s, 1H), 7.81 (m, 2H), 7.69-7.65 (m, 2H), 7.46-7.41 (m, 2H), 7.22-7.17 (m, 1H), 7.10-7.03 (m, 4H), 5.48 (m, 1H), 2.40-2.23 (m, 8H), 2.11 (s, 3H), 2.00-1.97 (m, 2H), 1.62-1.47 (m, 6H).

[0314] cis I-10: LCMS (m/z): 511.0 [M+H].sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz): δ (ppm) 8.59 (s, 1H), 8.40 (s, 1H), 7.81 (m, 2H), 7.68-7.65 (m, 2H), 7.46-7.42 (m, 2H), 7.19 (t, J=7.2 Hz, 1H), 7.10-7.07 (m, 4H), 5.19 (m, 1H), 2.49 (m, 3H), 2.29-2.18 (m, 7H), 2.13 (s, 3H), 1.83 (m, 2H), 1.44-1.39 (m, 4H).

Example 11. Synthesis of Cis and Trans I-7

[0315] The synthesis of trans 7-(4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(3-phenoxyprop-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine and cis 7-(4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(3-phenoxyprop-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (I-7) was carried out according to the below scheme.

##STR00096## ##STR00097##

5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (1f)

[0316] A mixture of 7H-pyrrolo[2,3-d]pyrimidin-4-amine (1.6 g, 11.9 mmol) and NIS (2.7 g, 11.9 mmol) in DCE (100 mL) was stirred at 80° C. for 3 h. After the reaction was completed the mixture was concentrated and purified by silica gel (DCM/MeOH=10/1) to obtain 1f (yellow solid, 2.4 g, 77% yield). LCMS: 261 (M+H).sup.+.

5-iodo-7-(1,4-dioxaspiro[4.5]decan-8-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (2f)

[0317] A mixture of 5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (1.4 g, 5.38 mmol), 1,4-dioxaspiro[4.5]decan-8-yl 4-methylbenzenesulfonate (1.7 g, 5.38 mmol) and CS.sub.2CO.sub.3 (3.5 g, 10.76 mmol) in DMSO (20 mL) was stirred at 100° C. for 16 h. After the reaction was completed the mixture was concentrated and purified by silica gel (DCM/MeOH=10/1) to obtain title compound 2f (yellow oil, 300 mg, 14% yield). LCMS: 401 (M+H).sup.+.

5-(3-phenoxyprop-1-ynyl)-7-(1,4-dioxaspiro[4.5]decan-8-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (3f)

[0318] A mixture of 5-(4-carbamoyl-3-(2-morpholinoethoxy)phenyl)-N-(2-methyl-5-(3-(trifluoromethyl)phenylcarbamoyl)phenyl)nicotinamide (280 mg, 0.7 mmol), (prop-2-ynyloxy)benzene (185 mg, 1.4 mmol), Pd(PPh.sub.3).sub.4 (81 mg, 0.07 mmol) and TEA (141 mg, 1.4 mmol) in DMF (4 mL) was stirred at 85° C. for 2 h. After the reaction was completed the mixture was concentrated and purified by silica gel (DCM/MeOH=10/1) to obtain 3f (yellow oil, 250 mg, 68% yield). LCMS: 405 (M+H).sup.+.

4-(4-amino-3-(3-phenoxyprop-1-ynyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexanone (4f)

[0319] A mixture of 3-(3-phenoxyprop-1-ynyl)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (120 mg, 0.29 mmol) and concentrated HCl (2 mL) in acetone (4 mL) was stirred at rt for 2 h. After the reaction was completed the mixture was concentrated and purified by silica gel (DCM/MeOH=10/1) to obtain title compound 4f (yellow solid, 100 mg, 93% yield). LCMS: 361 (M+H).sup.+.

trans 7-(4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(3-phenoxyprop-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (trans I-7) and cis 7-(4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(3-phenoxyprop-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (cis I-7)

[0320] A mixture of 4-(4-amino-3-(3-phenoxyprop-1-ynyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexanone (90 mg, 0.25 mmol), morpholin (200 mg, 1.25 mmol) and NaBH(OAc).sub.3 (176 mg, 0.83 mmol) in DCE (5 mL) was stirred at rt for 16 h. After the reaction was completed, the reaction mixture was concentrated and purified by prep-HPLC (C18 column, CH.sub.3CN/H.sub.2O, containing 0.05% NH.sub.4HCO.sub.3) to get title compound trans I-7 (white solid, 4 mg, 8%) and cis I-7 (white solid, 15 mg, 13.5%). HPLC: 100% (@254 nm).

[0321] trans I-7: LCMS: 445 (M+H); .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.25 (s, 1H), 7.34 (t, J=7.8 Hz, 2H), 7.19 (s, 1H), 7.02 (t, J=8.1 Hz, 3H), 5.27 (s, 2H), 4.97 (s, 2H), 4.56 (s, 1H), 2.59 (d, J=60.1 Hz, 9H), 2.33 (s, 3H), 2.09 (d, J=28.1 Hz, 4H), 1.73 (d, J=13.8 Hz, 2H), 1.56-1.47 (m, 2H).

[0322] cis I-7: LCMS: 445 (M+H); .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.25 (s, 1H), 7.50-7.29 (m, 3H), 7.14-6.92 (m, 3H), 5.27 (s, 2H), 4.98 (s, 2H), 4.72 (s, 1H), 2.49 (s, 6H), 2.31 (s, 3H), 2.26 (s, 1H), 2.05 (dd, J=24.2, 12.6 Hz, 4H), 1.85-1.36 (m, 6H).

Example 12. Synthesis of Cis and Trans I-8

[0323] The synthesis of trans 1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(3-phenoxyprop-1-yn-1-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (trans I-8) and cis 1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(3-phenoxyprop-1-yn-1-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (cis I-8) was carried out following the same procedure as trans I-7 and cis I-7, but starting with 3-bromo-1H-pyrazolo[3,4-d]pyrimidin-4-amine instead.

[0324] trans I-8: LCMS: 446.1 [M+H].sup.+; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.29 (s, 1H), 7.44-7.29 (m, 2H), 7.03 (dd, J=17.7, 9.9 Hz, 3H), 5.34 (br, 2H), 5.03 (s, 2H), 4.68 (brs, 1H), 2.75-2.40 (m, 8H), 2.31 (s, 3H), 2.15-2.06 (m, 6H), 1.53-1.38 (m, 3H).

[0325] cis I-8: LCMS: 446.2 [M+H].sup.+; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.31 (s, 1H), 7.40-7.35 (m, 2H), 7.10-7.04 (m, 3H), 5.40 (s, 2H), 5.04 (s, 2H), 4.88-4.80 (m, 1H), 2.80-2.45 (m, 6H), 2.44-2.22 (m, 7H), 2.14-2.08 (m, 2H), 1.80-1.45 (m, 5H).

Example 13. Synthesis of Cis and Trans I-2

[0326] The synthesis of 3-(4-(4-methylpiperazin-1-yl)cyclohexyl)-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine (I-2) was carried out according to the following scheme.

##STR00098## ##STR00099## ##STR00100##

N-((3-chloropyrazin-2-yl)methyl)-4-oxocyclohexane-1-carboxamide (1g)

[0327] A mixture of (3-chloropyrazin-2-yl)methanamine (3.0 g, 21.0 mmol), 4-oxocyclohexane-1-carboxylic acid (3.8 g, 27.3 mmol), HOBT (4.2 g, 31.5 mmol), EDCI (6.0 g, 31.5 mmol), DIPEA (2.0 mL) and DCM (50 mL) was stirred at r.t. for 16 h. After this time, the reaction mixture was washed with brine (50 mL×2), extracted with DCM. The combined organic extract was dried with Na.sub.2SO.sub.4, filtered, and purified by silica-gel (MeOH/DCM=1/20) to obtain 1g (white solid, 1.7 g, yield 30%). LCMS (m/z): 268.0 [M+H].sup.+.

4-(8-chloroimidazo[1,5-a]pyrazin-3-yl)cyclohexan-1-one (2g)

[0328] POCl.sub.3 (4.0 mL) was dropped to the mixture of 1 (1.7 g, 6.3 mmol), DMF (1.0 mL) and CH.sub.3CN (60 mL) at 0° C., then stirred for 3 h, after completion, quenched with DIPEA, concentrated, washed with H.sub.2O (50 mL×2), extracted with EA, dried with Na.sub.2SO.sub.4, filtered, removed the solvent to obtain 2 (yellow solid, 1.0 g, yield 66%). LCMS (m/z): 249.9 [M+H].sup.+.

4-(8-chloro-1-iodoimidazo[1,5-a]pyrazin-3-yl)cyclohexan-1-one (3g)

[0329] The mixture of 2 (0.5 g, 2.0 mmol), NIS (0.54 g, 2.4 mmol) and DMF (8.0 mL) was stirred at 55° C. for 16 h, extracted with ethyl acetate, washed with brine (50 mL×5), dried with Na.sub.2SO.sub.4, filtered, removed the solvent, purified by silica-gel (EA/PE=8/1) to obtain 3 (yellow solid, 0.5 g, yield 66%). LCMS (m/z): 375.8 [M+H].sup.+.

4-(8-chloro-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)cyclohexan-1-one (4g)

[0330] A mixture of 3g (0.43 g, 1.1 mmol), 4,4,5,5-tetramethyl-2-(4-phenoxyphenyl)-1,3,2-dioxaborolane (0.34 g, 1.1 mmol), Cs.sub.2CO.sub.3 (0.40 g, 1.25 mmol), Pd(dppf)Cl.sub.2 (0.093 g, 0.11 mmol), DMF (0.1 mL) and Toluene (8.0 mL) was stirred at 70° C. for 16 h. After this time, the reaction mixture was extracted with ethyl acetate, and the combined organic extracts were washed with brine (50 mL×2), dried with Na.sub.2SO.sub.4, filtered, concentrated, purified by silica-gel (EA/PE=1/1) to obtain 4g (yellow solid, 0.37 g, yield 77%). LCMS (m/z): 417.9 [M+H].sup.+.

8-chloro-3-((1s,4s)-4-(4-methylpiperazin-1-yl)cyclohexyl)-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazine (5g)

[0331] A mixture of 4g (0.3 g, 0.72 mmol), 1-methylpiperazine (2 mL), CH.sub.3COOH (0.1 mL) and DCE (10 mL) was stirred at rt for 3 h. Then NaBH(OAc).sub.3 (0.6 g, 2.9 mmol) was added and this mixture was stirred overnight. After this time, the reaction mixture was washed with brine (50 mL×2), extracted with DCM, and the organic phase was dried with Na.sub.2SO.sub.4, filtered, concentrated, purified by Pre-TLC (MeOH/DCM=1/8), obtained 5 (yellow solid, 0.15 g, yield 41%). LCMS (m/z): 502.0 [M+H].sup.+.

N,N-bis(4-methoxybenzyl)-3-((1s,4s)-4-(4-methylpiperazin-1-yl)cyclohexyl)-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine (6g)

[0332] A mixture of 5g (0.12 g, 0.24 mmol), bis(4-methoxybenzyl)amine (0.123 g, 0.48 mmol), DIPEA (1.0 mL) and 2-BuOH (4.0 mL) was stirred at 120° C. for 48 h. After this time, the reaction mixture was concentrated and purified by silica-gel (MeOH/DCM=1/10) to obtain 6g (yellow oil, 50 mg, yield 29%). LCMS (m/z): 723.4 [M+H].sup.+.

3-((1s,4s)-4-(4-methylpiperazin-1-yl)cyclohexyl)-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine (I-2)

[0333] A mixture of 6g (40 mg, 0.05 mmol), TFA (1.0 mL), triflic acid (0.5 mL) and DCE (2.0 mL) was stirred at 50° C. for 16 h. After this time, the reaction mixture was concentrated, purified by prep-TLC (MeOH/DCM=1/8) and prep-HPLC (C18 column, CH.sub.3CN/H.sub.2O, containing 0.05% NH.sub.4HCO.sub.3) to obtain I-2 (white solid, 0.5 mg). LCMS (m/z): 483.0 [M+H].sup.+; 1H NMR (DMSO, 400 MHz): δ 7.67-7.51 (m, 3H), 7.43 (t, J=7.6 Hz, 2H), 7.18 (t, J=7.2 Hz, 1H), 7.13-7.09 (m, 4H), 7.02 (d, J=4.8 Hz, 1H), 6.03 (s, 2H), 3.64-3.51 (m, 8H), 2.02-1.98 (m, 4H), 1.73-1.66 (m, 4H), 1.23 (s, 5H).

EQUIVALENTS AND SCOPE

[0334] In the claims articles such as “a,” “an,” and “the” may mean one or more than one unless indicated to the contrary or otherwise evident from the context. Claims or descriptions that include “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 evident 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.

[0335] Furthermore, the present disclosure encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims is introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and any element(s) can be removed from the group. It should it be understood that, in general, where the present disclosure, or aspects of the present disclosure, is/are referred to as comprising particular elements and/or features, certain embodiments of the present disclosure or aspects of the present disclosure consist, or consist essentially of, such elements and/or features. For purposes of simplicity, those embodiments have not been specifically set forth in haec verba herein. It is also noted that the terms “comprising” and “containing” are intended to be open and permits the inclusion of additional elements or steps. Where ranges are given, endpoints are included. Furthermore, unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or sub-range within the stated ranges in different embodiments of the present disclosure, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

[0336] This application refers to various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. If there is a conflict between any of the incorporated references and the instant specification, the specification shall control. In addition, any particular embodiment of the present disclosure that falls within the prior art may be explicitly excluded from any one or more of the claims. Because such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. Any particular embodiment of the present disclosure can be excluded from any claim, for any reason, whether or not related to the existence of prior art.

[0337] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments described herein. The scope of the present embodiments described herein is not intended to be limited to the above Description, but rather is as set forth in the appended claims. Those of ordinary skill in the art will appreciate that various changes and modifications to this description may be made without departing from the spirit or scope of the present disclosure, as defined in the following claims.