Reverse thiazine kinase inhibitors

Abstract

Disclosed is a class of kinase inhibitors of formula (I). Related pharmaceutical compositions and methods of using the kinase inhibitors are also disclosed.

Description

REVERSE THIAZINE KINASE INHIBITORSCROSS-REFERENCE TO RELATED APPLICATION

[0001] This patent application claims the benefit of U.S. Provisional Patent Application No. 63 / 672,577, filed July 17, 2024, which is incorporated by reference in its entirety herein.STATEMENT REGARDINGFEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] This invention was made with Government support under project number Z01ZIABC011744 by the National Institutes of Health, National Cancer Institute, project number Z01ZIABC011471 by the National Institutes of Health, National Cancer Institute, and project number Z01ZIABC011854 by the Cancer Moonshot NCI Program for Natural Product Discovery. The Government has certain rights in this invention.BACKGROUND OF THE INVENTION

[0003] Mammals have enzymes called kinases that are associated with cell functions such as cell signaling, metabolism, and division. Some kinases have been found to be more active in certain types of cancers. Blocking the kinases associated with cancer growth may provide therapeutic advantages to those suffering from cancer. Given that cancer is currently a major health concern and that there is a lack of effective treatments against all cancers, there is an urgent need to identify new kinase inhibitors to treat cancers. There is also an urgent need to identify kinase inhibitors associated with non-cancer pathologies (e.g., infections) in order to treat conditions and disorders associated with non-cancer pathologies.

[0004] The Molecular Targets Program of the United States’ National Cancer Institute (NCI) completed a screen of -150,000 pre-fractionated natural products from the NCI Program for Natural Product Discovery (NPNPD) (Thornburg, et al., ACS Chem. Biol., 13: 2484-2497 (2018)). A class of active compounds identified were isolated from the marine organism Aplidium sp. These compounds, named Aplithianines A & B, were shown to potently inhibit both (1) oncogenic gene fusion DNAJB1-PRKACA and (2) wild type protein kinase A (PKA) at nanomolar concentrations. Aplithianine A was shown to potently and selectively inhibit a broad range of kinases, not just J-PKAcα or PKA, broadening its potential utility. Further kinetic analysis showed that Aplithianine A was a competitiveinhibitor of kinases, competing with ATP for binding to J-PKAcα. Additional structural studies showed that aplithianine A bound to the catalytic pocket in J-PKAcα where ATP normally binds, further proving the competitive mechanism of inhibition and providing structural insights for further synthetic modification of this compound class. The aplithianine structural class is a group of potent kinase inhibitors with broad potential applicability to numerous kinases of importance, e.g., for cancer chemotherapy. For example, gene fusions (a genetic lesion ligating two normally non-adjacent portions of the genome next to one another) were one of the earliest recognized biomarkers of cancer. Approximately 20% of all solid malignancies have at least one identifiable gene fusion. The experience with the BCR-ABL1 kinase inhibitor imatinib (Savage, et al., N. Engl. J. Med., 346(9): 683-93 (2002)), and a continuing emphasis on precision medicine suggests that focusing on gene fusion associated pharmaceutical development could produce disease specific medicines. One such fusion is the recently identified DNAJB1-PRKACA oncogenic gene fusion associated with fibrolamellar hepatocellular carcinoma (FL-HCC) (Honeyman, et al., Science, 343: 1010-14 (2014) and Kastenhuber, et al., PNAS USA, 114: 13076-84 (2017)). Among liver cancers, FL-HCC is unusually tragic in that its patient population is young (<35 years of age) and lacks any successful disease specific chemotherapeutic regime, with a 5 year survival rate of only approximately 34% (Riggle, et al., Pediatr. Blood Cancer, 63: 1163-7 (2016)). The biological understanding of FL-HCC improved in 2014 when for the first time it was shown that all FL-HCC patients carried an in-frame intrachromosomal gene fusion between the first exon of the gene encoding the Heat Shock Protein 40 (HSP40) family member DNAJB1 and the second exon of the gene for the adenosine 3’,5’-monophosphate (cAMP)–dependent PKA catalytic subunit alpha, PRKACA (Honeyman, et al., Science, 343: 1010-14 (2014)). The DNAJB1-PRKACA gene fusion produces an enzymatically active chimeric protein J-PKAcα. Studies have demonstrated that PKA activity was required for tumor formation. Equivalent expression of PKAcα or expression of a kinase-dead version of the oncogenic fusion protein is not sufficient for transformation and the tumorigenicity of J- PKAcα is dependent on its kinase activity (Kastenhuber, et al., PNAS USA, 114: 13076-84 (2017)). J-PKAcα fusion complexes may present novel small molecule binding sites which can be exploited for the treatment of FL-HCC (Tomasini, et al., Scientific Reports, 8: 720 (2018); Cheung, et al., PNAS USA, 112: 1374-79 (2015); and Averill, et al., J. Cell Biochem., 120: 13783-91 (2019)).Thus, there remains a need to develop novel small molecules that can act as both inhibitors and activators of J-PKAcα. There also exists a need for methods of treatment using such inhibitors and activators of J-PKAcα. The invention provides such small molecules that can act as both inhibitors and activators of J-PKAcα and methods. These, and other advantages of the invention, as well as additional inventive features, will be apparent from the description of the invention provided herein. BRIEF SUMMARY OF THE INVENTION In some aspects, the invention provides compounds of formula (I):wherein X1, X2, X3,R1, and E are defined herein, or a pharmaceutically acceptable salt thereof. The invention also provides pharmaceutical compositions comprising compounds of formula (I). The invention further provides methods of inhibiting kinase activity in a subject, methods of suppressing the immune system in a subject, methods of preventing organ rejection in a subject, methods of treating cancer in a subject, methods of treating diabetic neuropathic pain in a subject, methods of treating malaria in a subject, and / or methods of treating an infection associated with a protozoa in a subject comprising administering to the subject compounds or pharmaceutical compositions of aspects of the present invention.BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS FIG.1 is a graph showing the normalized % JPKAcα Activity curve for Compound D. FIG.2 is a graph showing the normalized % JPKAcα Activity curve for Compound E. FIG.3 is a graph showing the normalized % JPKAcα Activity curve for Compound F. FIG.4 is a graph showing the normalized % JPKAcα Activity curve for Compound G. FIG.5 is a graph showing the normalized % JPKAcα Activity curve for Compound H. FIG.6 is a graph showing the normalized % JPKAcα Activity curve for Compound I. FIG.7 is a graph showing the normalized % JPKAcα Activity curve for Compound J. FIG.8 is a graph showing the normalized % JPKAcα Activity curve for Compound K. FIG.9 is a graph showing the normalized % JPKAcα Activity curve for Compound L. FIG.10 is a graph showing the HCT-116 cell viability curve for Compound D. FIG.11 is a graph showing the HCT-116 cell viability curve for Compound H. FIG.12 is a graph showing the HCT-116 cell viability curve for Compound I. FIG.13 is a graph showing the HCT-116 cell viability curve for Compound L. FIG.14 is a graph showing the normalized homogeneous time resolved fluorescence (HTRF) percentage of the Western Blot study for Compound J. HillSlope was -1.683, and EC50was 5.560 x 10-6M. FIG.15 is a graph showing the normalized homogeneous time resolved fluorescence (HTRF) percentage of the Western Blot study for Compound L. HillSlope was -1.223, and EC50was 2.821 x 10-7M. FIG.16 is a ribbon diagram showing the structure of the co-crystallized complex of J-PKAcα and Compound L. FIG.17 is a diagram showing J-PKAcα residues that mediate the binding affinity of Compound L.FIG.18 is a graph showing the dose-response study of Compound I in a malaria asexual assay. FIG.19 is a bar graph showing Compounds B and J inhibiting CLK-dependent RNA splicing. DETAILED DESCRIPTION OF THE INVENTION In some aspects, the invention provides a compound of formula (I)whereinis a single or double bond, X1and X2are each independently CH, CR4, or N; X3is S, S=O, or S(=O)2; R1is H or –NR2R3; R2is H, C1-C3alkyl, or an aryl; R3is H, C1-C3alkyl, or an aryl; R4is C1-C3alkyl; and E is C3-C8heterocycloalkyl, C8-C10bicycloalkyl, C4-C10biheterocycloalkyl, aryl, or heteroaryl, each of which is optionally substituted with one or more substituents selected from C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, trifluoromethyl, C1-C6alkoxy, arylated C1-C6alkoxy, trifluoromethylated C1-C6alkoxy, -C(O)OH, -C(O)O-(C1-C6alkyl), -C(O)-(C1-C6alkyl), -(C1-C3alkyl)-C(O)OH, -(C1-C3alkyl)-C(O)O-(C1-C6alkyl), -(C1-C3alkyl)-C(O)-(C1- C6alkyl), -NH-C(O)OH, -NH-C(O)O-(C1-C6alkyl), -NH-C(O)-(C1-C6alkyl), -(C1-C3alkyl)-NH-C(O)OH, -(C1-C3alkyl)-NH-C(O)O-(C1-C6alkyl), -(C1-C3alkyl)-NH-C(O)-(C1-C6alkyl), -(C1-C6alkyl)-O-(C1-C6alkyl), -O-(C1-C6alkyl)-O-(C1-C6alkyl), -(C1-C6alkyl)-OH, - (C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-COOH, -(C1-C6alkyl)-C(O)O-(C1-C6alkyl), -(C1-C6alkyl)-(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-NH2, -(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)- (C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)-heterocycloalkyl, -(C1-C6alkyl)-heterocycloalkyl- (C1-C6alkyl), -(C1-C6alkyl)-C(O)-heterocycloalkyl, -(C1-C6alkyl)-C(O)-heterocycloalkyl- (C1-C6alkyl), halo, nitro, hydroxy, amino, C1-C6alkylamino, di-C1-C6alkyl-amino; -NH- aryl, -NH-heterocycloalkyl, -NH-heterocycloalkyl-(C1-C6alkyl), -C1-C6haloalkyl, C3-C8cycloalkyl, C3-C8heterocycloalkyl, fused C3-C8cycloalkyl, spiro C3-C8cycloalkyl, fused C3- C8heterocycloalkyl, spiro C3-C8heterocycloalkyl, aryl, heteroaryl, fused aryl, fused heteroaryl, -CN, -(C1-C3alkyl)-CN, carbonyl, amino acid, amino acid methyl ester, and a combination thereof, or a pharmaceutically acceptable salt thereof. In an aspect of the invention, the compound of formula (I) is of formula (Ia): ,or a pharmaceutically acceptable salt thereof. In an aspect of the invention, the compound of formula (I) is of formula (Ib):7or a pharmaceutically acceptable salt thereof.

[0034] In an aspect of the invention, the compound of formula (I) is of formula (Ic):or a pharmaceutically acceptable salt thereof.

[0035] In an aspect of the invention, the compound of formula (I) is of formula (Id):or a pharmaceutically acceptable salt thereof.In an aspect of the invention, the compound of formula (I) is of formula (Ie): ,or a pharmaceutically acceptable salt thereof. In an aspect of the invention, the compound of formula (I) is of formula (If): ,or a pharmaceutically acceptable salt thereof. In some embodiments (i.e., any of the aspects described herein) of the compound of formula (I), E is selected from:,each of which is optionally substituted with one or more substituents selected from C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, trifluoromethyl, C1-C6alkoxy, arylated C1-C6alkoxy, trifluoromethylated C1-C6alkoxy, -C(O)OH, -C(O)O-(C1-C6alkyl), -C(O)-(C1-C6alkyl), - (C1-C3alkyl)-C(O)OH, -(C1-C3alkyl)-C(O)O-(C1-C6alkyl), -(C1-C3alkyl)-C(O)-(C1-C6alkyl), -NH-C(O)OH, -NH-C(O)O-(C1-C6alkyl), -NH-C(O)-(C1-C6alkyl), -(C1-C3alkyl)- NH-C(O)OH, -(C1-C3alkyl)-NH-C(O)O-(C1-C6alkyl), -(C1-C3alkyl)-NH-C(O)-(C1-C6alkyl), -(C1-C6alkyl)-O-(C1-C6alkyl), -O-(C1-C6alkyl)-O-(C1-C6alkyl), -(C1-C6alkyl)-OH, - (C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-COOH, -(C1-C6alkyl)-C(O)O-(C1-C6alkyl), -(C1-C6alkyl)-(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-NH2, -(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)- (C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)-heterocycloalkyl, -(C1-C6alkyl)-heterocycloalkyl- (C1-C6alkyl), -(C1-C6alkyl)-C(O)-heterocycloalkyl, -(C1-C6alkyl)-C(O)-heterocycloalkyl- (C1-C6alkyl), halo, nitro, hydroxy, amino, C1-C6alkylamino, di-C1-C6alkyl-amino; -NH- aryl, -NH-heterocycloalkyl, -NH-heterocycloalkyl-(C1-C6alkyl), -C1-C6haloalkyl, C3-C8cycloalkyl, C3-C8heterocycloalkyl, fused C3-C8cycloalkyl, spiro C3-C8cycloalkyl, fused C3- C8heterocycloalkyl, spiro C3-C8heterocycloalkyl, aryl, heteroaryl, fused aryl, fused heteroaryl, -CN, -(C1-C3alkyl)-CN, carbonyl, amino acid, amino acid methyl ester, and a combination thereof. In some embodiments, E is a carbon bound C3-C8heterocycloalkyl, aryl, or heteroaryl, each of which is optionally substituted with one or more substituents selected from C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, trifluoromethyl, C1-C6alkoxy, arylated C1-C6alkoxy, trifluoromethylated C1-C6alkoxy, -C(O)OH, -C(O)O-(C1-C6alkyl), -C(O)-(C1-C6alkyl), -(C1-C3alkyl)-C(O)OH, -(C1-C3alkyl)-C(O)O-(C1-C6alkyl), -(C1-C3alkyl)-C(O)-(C1-C6alkyl), -NH-C(O)OH, -NH-C(O)O-(C1-C6alkyl), -NH-C(O)-(C1-C6alkyl), -(C1-C3alkyl)- NH-C(O)OH, -(C1-C3alkyl)-NH-C(O)O-(C1-C6alkyl), -(C1-C3alkyl)-NH-C(O)-(C1-C6alkyl), -(C1-C6alkyl)-O-(C1-C6alkyl), -O-(C1-C6alkyl)-O-(C1-C6alkyl), -(C1-C6alkyl)-OH, - (C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-COOH, -(C1-C6alkyl)-C(O)O-(C1-C6alkyl), -(C1-C6alkyl)-(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-NH2, -(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)- (C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)-heterocycloalkyl, -(C1-C6alkyl)-heterocycloalkyl- (C1-C6alkyl), -(C1-C6alkyl)-C(O)-heterocycloalkyl, -(C1-C6alkyl)-C(O)-heterocycloalkyl- (C1-C6alkyl), halo, nitro, hydroxy, amino, C1-C6alkylamino, di-C1-C6alkyl-amino; -NH- aryl, -NH-heterocycloalkyl, -NH-heterocycloalkyl-(C1-C6alkyl), -C1-C6haloalkyl, C3-C8cycloalkyl, C3-C8heterocycloalkyl, fused C3-C8cycloalkyl, spiro C3-C8cycloalkyl, fused C3- C8heterocycloalkyl, spiro C3-C8heterocycloalkyl, aryl, heteroaryl, fused aryl, fused heteroaryl, -CN, -(C1-C3alkyl)-CN, carbonyl, amino acid, amino acid methyl ester, and a combination thereof. In some embodiments (i.e., any of the aspects described herein) of the compound of formula (I), E comprises at least one nitrogen atom (e.g., at least two nitrogen atoms or at least three nitrogen atoms). In certain embodiments, E contains exactly 1 nitrogen atom. In other embodiments, E contains exactly 2 nitrogen atoms. In some embodiments, E contains exactly 3 nitrogen atoms. In some embodiments (i.e., any of the aspects described herein) of the compound of formula (I), E comprises at least 3 (e.g., 3-10, 4-10, 5-10, 6-10, 7-10, 4-12, 5-12, 6-12, or 7-12) carbon atoms. In some embodiments (i.e., any of the aspects described herein) of the compound of formula (I), E contains only carbon, nitrogen, and hydrogen atoms, e.g., does not contain an oxygen atom. In some embodiments (i.e., any of the aspects described herein) of the compound of formula (I), E contains only carbon, nitrogen, oxygen, and hydrogen atoms. In some embodiments (i.e., any of the aspects described herein) of the compound of formula (I), E contains only carbon, nitrogen, sulfur, and hydrogen atoms. In some embodiments (i.e., any of the aspects described herein) of the compound of formula (I), is a single bond. In some embodiments (i.e., any of the aspects described herein) of the compound of formula (I), is a double bond.In some embodiments (i.e., any of the aspects described herein) of the compound of formula (I), R1is H. In some embodiments (i.e., any of the aspects described herein) of the compound of formula (I), X1and X2are not both N. In some embodiments (i.e., any of the aspects described herein) of the compound of formula (I), X1and X2are each independently CH or CR4. In some embodiments (i.e., any of the aspects described herein) of the compound of formula (I), X1and X2are each independently CH. In some embodiments, the compound of formula (I) is:14or a pharmaceutically acceptable salt thereof

[0051] Any of the compounds of formula (I) can exist as any suitable stereoisomer thereof For example, the invention provides enantiomers and diastereomers of any of the compounds disclosed herein. Alternatively, any of the compounds of formula (I) can exist as a racemic mixture and / or a mixture of diastereomers.

[0052] In any of the aspects of the compounds of formula (I), the term “alkyl” implies a straight-chain or branched alkyl substituent containing from, for example, from about 1 to about 6 carbon atoms, e.g., from about 1 to about 4 carbon atoms. Examples of alkyl groupinclude methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, n-hexyl, and the like. This definition also applies wherever “alkyl” occurs as part of a group, such as, e.g., in C3-C6cycloalkylalkyl, hydroxyalkyl, haloalkyl (e.g., monohaloalkyl, dihaloalkyl, and trihaloalkyl), cyanoalkyl, aminoalkyl, alkylamino, dialkylamino, alkylaminoalkyl, dialkylaminoalkyl, arylcarbonylalkyl (-(alkyl)C(O)aryl), arylalkyl, etc. The alkyl can be substituted or unsubstituted, as described herein. Even in instances in which the alkyl is an alkylene chain (e.g., -(CH2)n-), the alkyl group can be substituted or unsubstituted. In any of the aspects of the compounds of formula (I), the term “alkenyl,” as used herein, means a linear alkenyl substituent containing from, for example, about 2 to about 6 carbon atoms (branched alkenyls are about 3 to about 6 carbon atoms), e.g., from about 3 to about 5 carbon atoms (branched alkenyls are about 3 to about 6 carbon atoms). In accordance with an aspect of the invention, the alkenyl group is a C2-C4alkenyl. Examples of alkenyl group include ethenyl, allyl, 2-propenyl, 1-butenyl, 2-butenyl, 1-pentenyl, 2- pentenyl, 3-pentenyl, 1-hexenyl, and the like. The alkenyl can be substituted or unsubstituted, as described herein. In any of the aspects of the compounds of formula (I), the term “cycloalkyl,” as used herein, means a cyclic alkyl moiety containing from, for example, 3 to 6 carbon atoms or from 5 to 6 carbon atoms. Examples of such moieties include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like. The cycloalkyl may contain a carbonyl such that there is an exocylclic (=O) group. The cycloalkyl can be substituted or unsubstituted, as described herein. The cycloalkyl may also be fused to a neighboring substituent (e.g., a cycloalkyl, a heterocycloalkyl, an aryl, or a heteroaryl), i.e, sharing two atoms and a bond with a neighboring substituent. In any of the aspects of the compounds of formula (I), the term “aryl” refers to a mono, bi, or tricyclic carbocyclic ring system having one, two, or three aromatic rings, for example, phenyl, naphthyl, anthracenyl, or biphenyl. The term “aryl” refers to an unsubstituted or substituted aromatic carbocyclic moiety, as commonly understood in the art, and includes monocyclic and polycyclic aromatics such as, for example, phenyl, biphenyl, naphthyl, anthracenyl, pyrenyl, and the like. An aryl moiety generally contains from, for example, 6 to 30 carbon atoms, from 6 to 18 carbon atoms, from 6 to 14 carbon atoms, or from 6 to 10 carbon atoms. It is understood that the term aryl includes carbocyclic moieties that are planar and comprise 4n+2 π electrons, according to Hückel’s Rule, wherein n = 1, 2,or 3. This definition also applies wherever “aryl” occurs as part of a group, such as, e.g., in haloaryl (e.g., monohaloaryl, dihaloaryl, and trihaloaryl), arylalkyl, etc. The aryl can be substituted or unsubstituted, as described herein. The aryl may also be fused to a neighboring substituent (e.g., a cycloalkyl, a heterocycloalkyl, an aryl, or a heteroaryl), i.e., sharing two atoms and a bond with a neighboring substituent.

[0056] In any of the aspects of the compounds of formula (I), the term “heteroaryl” refers to aromatic 5 or 6 membered monocyclic groups, 9 or 10 membered bicyclic groups, and 11 to 14 membered tricyclic groups which have at least one heteroatom (O, S, or N) in at least one of the rings. Each ring of the heteroaryl group containing a heteroatom can contain one or two oxygen or sulfur atoms and / or from one to four nitrogen atoms provided that the total number of heteroatoms in each ring is four or less and each ring has at least one carbon atom. The fused rings completing the bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated, or unsaturated. The nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen atoms may optionally be quatemized. Heteroaryl groups which are bicyclic or tricyclic must include at least one fully aromatic ring but the other fused ring or rings may be aromatic or non-aromatic. The heteroaryl group may be attached at any available nitrogen or carbon atom of any ring. Illustrative examples of heteroaryl groups are pyridinyl, pyridazinyl, pyrimidyl, pyrazinyl, benzimidazolyl, triazinyl, imidazolyl, (1,2,3)- and (l,2,4)-triazolyl, pyrazinyl, tetrazolyl, furyl, pyrrolyl, thienyl, isothiazolyl, thiazolyl, isoxazolyl, and oxadiazolyl. The heteroaryl can be substituted or unsubstituted, as described herein. The heteroaryl may also be fused to a neighboring substituent (e.g., a cycloalkyl, a heterocycloalkyl, an aryl, or a heteroaryl), i.e., sharing two atoms and a bond with a neighboring substituent.

[0057] In any of the aspects of the compounds of formula (I), the term “heterocycloalkyl” means a stable, saturated, or partially unsaturated monocyclic, bicyclic, and spiro ring system containing 3 to 7 ring members of carbon atoms and other atoms selected from nitrogen, sulfur, and / or oxygen. In an aspect, a heterocycloalkyl is a 5, 6, or 7-membered monocyclic ring and contains one, two, or three heteroatoms selected from nitrogen, oxygen, and sulfur. The heterocycloalkyl may be attached to the parent structure through a carbon atom or through any heteroatom of the heterocycloalkyl that results in a stable structure. Alternatively, or additionally, the heterocycloalkyl may contain a carbonyl such that there is an exocylclic (=0) group. In other words, as used herein when referring to a substituent, the term “carbonyl” refers to a (=0) group. Examples of such heterocycloalkyl rings areisoxazolyl, thiazolinyl, imidazolidinyl, piperazinyl, homopiperazinyl, pyrrolyl, pyrrolinyl, pyrazolyl, pyranyl, piperidyl, oxazolyl, and morpholinyl. The heterocycloalkyl can be substituted or unsubstituted, as described herein. In any of the aspects of the compound of formula (I), the term “C8-C10bicycloalkyl” refers to an aryl group fused to a cycloalkyl group, as described herein. In any of the aspects of the compound of formula (I), the term “C4-C10biheterocycloalkyl” refers to an aryl group fused to a heterocycloalkyl group, a heteroaryl group fused to a heterocycloalkyl group, or a heteroaryl group fused to a cycloalkyl group, as described herein. In any of the aspects of the compound of formula (I), the term “fused” means that the indicated ring system shares a bond with the ring system to which it is attached. In any of the aspects of the compound of formula (I), the term “spiro” means that the indicated ring system shares an atom with the ring system to which it is attached. In any of the aspects of the compounds of formula (I), the term “hydroxy” refers to the group –OH. In any of the aspects of the compounds of formula (I), the term “cyano” refers to the group –CN, whereas the term “thiocyano” refers to -SCN. In any of the aspects of the compounds of formula (I), the terms “alkoxy” and “cycloalkyloxy” embrace linear or branched alkyl and cycloalkyl groups, respectively, that are attached to a divalent oxygen or form a divalent oxygen atom by way of their attachment. The alkyl and cycloalkyl groups are the same as described herein. In any of the aspects of the compounds of formula (I), the term “halo” refers to a halogen selected from fluorine, chlorine, bromine, and iodine. In any of the aspects of the compounds of formula (I), the term “carboxylato” refers to the group -C(O)OH. In any of the aspects of the compounds of formula (I), the term “amino” refers to the group –NH2. The term “alkylamino” refers to –NHR, whereas the term “dialkylamino” refers to –NRR’. R and R' are the same or different and each is a substituted or unsubstituted alkyl group, as described herein. In any of the aspects of the compounds of formula (I), the term “amido” refers to the group -C(O)NRR’, which R and R’ are the same or different and each is hydrogen or a substituted or unsubstituted alkyl group, as described herein.In any of the aspects of the compounds of formula (I), the term “phosphonato” refers to the group -P(O)(OR)2, which R is hydrogen or a substituted or unsubstituted alkyl group, as described herein. In any of the aspects of the compounds of formula (I) and formula (II), the term “amino acid” refers to any amino acid selected from alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. In some embodiments, when used as a substituent, the amino acid is alanine or glycine. In certain embodiments, when used as a substituent, the amino acid is nitrogen bound. In any of the aspects of the compounds of formula (I) and formula (II), the term “amino acid methyl ester” refers to any amino acid described herein that has been converted to the methyl ester. In any of the aspects of the compounds of formula (I), any substituent that is not hydrogen (e.g., C1-C6alkyl, C2-C6alkenyl, C3-C6cycloalkyl, C3-C6cycloalkylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, or heterocycloalkylalkyl) can be an optionally substituted moiety. The substituted moiety typically comprises at least one substituent (e.g., 1, 2, 3, 4, 5, 6, etc.) in any suitable position (e.g., 1-, 2-, 3-, 4-, 5-, or 6- position, etc.). When an aryl group is substituted with a substituent, e.g., halo, amino, alkyl, OH, alkoxy, and others, the aromatic ring hydrogen is replaced with the substituent and this can take place in any of the available hydrogens, e.g., 2, 3, 4, 5, and / or 6-position wherein the 1-position is the point of attachment of the aryl group in the compound of the present invention. Suitable substituents include, e.g., halo, alkyl, alkenyl, hydroxy, nitro, cyano, amino, alkylamino, alkoxy, aryloxy, aralkoxy, carboxyl, carboxyalkyl, carboxyalkyloxy, amido, alkylamido, haloalkylamido, aryl, heteroaryl, heterocycloalkyl, cycloalkyl, fused aryl, fused heteroaryl, fused heterocycloalkyl, and fused cycloalkyl, each of which is described herein. In some embodiments, the one or more substituents is selected from C1-C6alkyl, C2- C6alkenyl, C2-C6alkynyl, trifluoromethyl, C1-C6alkoxy, arylated C1-C6alkoxy, trifluoromethylated C1-C6alkoxy, -(C1-C6alkyl)-OH, -(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-COOH, -(C1-C6alkyl)-NH2, -(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)-(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl) heterocycloalkyl, halo, nitro, hydroxy, amino, C1-C6alkylamino, di-C1-C6alkyl-amino; -NH-aryl, C1-C6haloalkyl, C3-C8cycloalkyl, C3-C8heterocycloalkyl, fused C3-C8cycloalkyl, fused C3-C8heterocycloalkyl, aryl, heteroaryl, fused aryl, fused heteroaryl, -CN, -(C1-C3alkyl)-CN,carbonyl, and a combination thereof. In some instances, the substituent is at least one alkyl, halo, and / or haloalkyl (e.g., 1 or 2). In any of the aspects of the compounds of formula (I), whenever a range of the number of atoms in a structure is indicated (e.g., a C1-12, C1-8, C1-6, C1-4, etc.), it is specifically contemplated that any sub-range or individual number of carbon atoms falling within the indicated range also can be used. Thus, for instance, the recitation of a range of 1-8 carbon atoms (e.g., C1-C8), 1-6 carbon atoms (e.g., C1-C6), 1-4 carbon atoms (e.g., C1-C4), 1-3 carbon atoms (e.g., C1-C3), or 2-8 carbon atoms (e.g., C2-C8) as used with respect to any chemical group (e.g., alkyl, cycloalkyl, etc.) referenced herein encompasses and specifically describes 1, 2, 3, 4, 5, 6, 7, and / or 8 carbon atoms, as appropriate, as well as any sub-range thereof (e.g., 1-2 carbon atoms, 1-3 carbon atoms, 1-4 carbon atoms, 1-5 carbon atoms, 1-6 carbon atoms, 1-7 carbon atoms, 1-8 carbon atoms, 2-3 carbon atoms, 2-4 carbon atoms, 2-5 carbon atoms, 2-6 carbon atoms, 2-7 carbon atoms, 2-8 carbon atoms, 3-4 carbon atoms, 3-5 carbon atoms, 3-6 carbon atoms, 3-7 carbon atoms, 3-8 carbon atoms, 4-5 carbon atoms, 4-6 carbon atoms, 4-7 carbon atoms, 4-8 carbon atoms, etc., as appropriate). In any of the aspects of the compounds of formula (I), the phrase “salt” or “pharmaceutically acceptable salt” is intended to include nontoxic salts synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two. For example, an inorganic acid (e.g., hydrochloric acid, sulfuric acid, phosphoric acid, or hydrobromic acid), an organic acid (e.g., oxalic acid, malonic acid, citric acid, fumaric acid, lactic acid, malic acid, succinic acid, tartaric acid, acetic acid, trifluoroacetic acid, gluconic acid, ascorbic acid, methylsulfonic acid, or benzylsulfonic acid), an inorganic base (e.g., sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, or ammonium hydroxide), an organic base(e.g., methylamine, diethylamine, triethylamine, triethanolamine, ethylenediamine, tris(hydroxymethyl)methylamine, guanidine, choline, or cinchonine), or an amino acid (e.g., lysine, arginine, or alanine) can be used. Generally, nonaqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are typical. Lists of suitable salts are found in Remington’s Pharmaceutical Sciences, 18th ed., Mack Publishing Company, Easton, PA, 1990, p.1445, and Journal of Pharmaceutical Science, 66, 2-19 (1977). For example, theycan be a salt of an alkali metal (e.g., sodium or potassium), alkaline earth metal (e.g., calcium), or ammonium of salt. In an aspect, the salt is a trifluoroacetate salt.Pharmaceutical Compositions

[0074] An aspect of the invention provides pharmaceutical compositions comprising a compound of the present invention. The pharmaceutical compositions contain a pharmaceutically acceptable carrier. For example, the pharmaceutical composition of the present invention can comprise a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0075] In an aspect, the compound has a purity of at least about 85% (e.g., at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, at least about 99.6%, at least about 99.7%, at least about 99.8%, or at least about 99.9%).

[0076] The pharmaceutical composition can comprise a compound of the present invention (e.g., a compound of formula (I)) in combination with one or more other pharmaceutically active agents or drugs, such as a chemotherapeutic agent, e.g., a topoisomerase I inhibitor, asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracil, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab, vinblastine, vincristine, etc.

[0077] Preferably, the carrier is a pharmaceutically acceptable carrier. With respect to pharmaceutical compositions, the carrier can be any of those conventionally used and is limited only by chemico-physical considerations, such as solubility and lack of reactivity with the active compound(s), and by the route of administration. The pharmaceutically acceptable carriers described herein, for example, vehicles, adjuvants, excipients, and diluents, are well-known to those skilled in the art and are readily available to the public. It is preferred that the pharmaceutically acceptable carrier be one which is chemically inert to the active agent(s) and one which has no detrimental side effects or toxicity under the conditions of use.

[0078] The choice of carrier will be determined in part by the particular compounds, as well as by the particular method used to administer the compounds. Accordingly, there are a variety of suitable formulations of the pharmaceutical composition of the invention. The compounds, a pharmaceutically acceptable salt thereof, can be administered in any suitablemanner (e.g., orally, intravenously, intramuscularly, intrathecally, subcutaneously, sublingually, buccally, rectally, vaginally, by ocular route, by otic route, nasally, by inhalation, by nebulization, topically, systemically, transdermally, or a combination thereof). In an embodiment, the pharmaceutical composition of the invention is administered orally.

[0079] The following formulations for administration are exemplary and are in no way limiting. More than one route can be used to administer the compounds, and in certain instances, a particular route can provide a more immediate and more effective response than another route.

[0080] Formulations suitable for administration include aqueous and non-aqueous, isotonic sterile injection solutions, which can contain anti-oxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. The compounds can be administered in a physiologically acceptable diluent in a pharmaceutical carrier, such as a sterile liquid or mixture of liquids, including water, saline, aqueous dextrose and related sugar solutions, an alcohol, such as ethanol or hexadecyl alcohol, a glycol, such as propylene glycol or polyethylene glycol, dimethylsulfoxide, glycerol, ketals such as 2,2-dimethyl-l,3-dioxolane- 4-methanol, ethers, poly(ethyleneglycol) 400, oils, fatty acids, fatty acid esters or glycerides, or acetylated fatty acid glycerides with or without the addition of a pharmaceutically acceptable surfactant, such as a soap or a detergent, suspending agent, such as pectin, carbomers, methylcellulose, hydroxypropylmethylcellulose, or carboxymethylcellulose, or emulsifying agents and other pharmaceutical adjuvants.

[0081] Oils, which can be used in formulations include petroleum, animal, vegetable, or synthetic oils. Specific examples of oils include peanut, soybean, sesame, cottonseed, com, olive, petrolatum, and mineral. Suitable fatty acids for use in formulations include oleic acid, stearic acid, and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters.

[0082] Suitable soaps for use in formulations include fatty alkali metal, ammonium, and triethanolamine salts, and suitable detergents include (a) cationic detergents such as, for example, dimethyl dialkyl ammonium halides, and alkyl pyridinium halides, (b) anionic detergents such as, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates, (c) nonionic detergents such as, for example, fatty amine oxides, fatty acid alkanolamides, and polyoxyethylenepolypropylene copolymers,(d) amphoteric detergents such as, for example, alkyl-P-aminopropionates, and 2-alkyl- imidazoline quaternary ammonium salts, and (e) mixtures thereof.

[0083] The formulations will typically contain from about 0.5% to about 25% by weight of the compounds in solution. Preservatives and buffers may be used. In order to minimize or eliminate irritation at the site of injection, such compositions may contain one or more nonionic surfactants having a hydrophile-lipophile balance (HLB) of from about 12 to about 17. The quantity of surfactant in such formulations will typically range from about 5% to about 15% by weight. Suitable surfactants include polyethylene glycol sorbitan fatty acid esters, such as sorbitan monooleate and the high molecular weight adducts of ethylene oxide with a hydrophobic base, formed by the condensation of propylene oxide with propylene glycol. The formulations can be presented in unit-dose or multi-dose sealed containers, such as ampoules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid excipient, for example, water, for injections, immediately prior to use. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules, and tablets of the kind previously described. The requirements for effective pharmaceutical carriers for compositions are well-known to those of ordinary skill in the art (see, e.g., Lloyd et al. (eds.), Remington: The Science and Practice of Pharmacy, 22nd Ed., Pharmaceutical Press (2012)).

[0084] It will be appreciated by one of skill in the art that, in addition to the abovedescribed pharmaceutical compositions, the compounds of the invention can be formulated as inclusion complexes, such as cyclodextrin inclusion complexes, or liposomes.

[0085] For purposes of the invention, the amount or dose of the compounds administered should be sufficient to effect a desired response, e.g., a therapeutic or prophylactic response, in the subject (e.g., mammal) over a reasonable time frame. For example, the dose of the compounds should be sufficient to inhibit growth of a target cell or treat or prevent cancer in a period of from about 2 hours or longer, e.g., 12 to 24 or more hours, from the time of administration. In certain aspects, the time period could be even longer. The dose will be determined by the efficacy of the particular compounds and the condition of the subject (e.g., mammal or human), as well as the body weight of the subject (e.g., mammal or human) to be treated.

[0086] Many assays for determining an administered dose are known in the art. An administered dose may be determined in vitro (e.g., cell cultures) or in vivo (e.g., animal studies). For example, an administered dose may be determined by determining the IC50(thedose that achieves a half-maximal inhibition of symptoms), LD50(the dose lethal to 50% of the population), the ED50(the dose therapeutically effective in 50% of the population), and the therapeutic index in cell culture and / or animal studies. The therapeutic index is the ratio of LD50to ED50(i.e., LD50 / ED50). The dose of the compounds also will be determined by the existence, nature, and extent of any adverse side effects that might accompany the administration of a particular compound. Typically, the attending physician will decide the dosage of the compounds with which to treat each individual patient, taking into consideration a variety of factors, such as age, body weight, general health, diet, sex, compounds to be administered, route of administration, and the severity of the condition being treated. By way of example and not intending to limit the invention, the dose of the compounds can be about 0.001 to about 1000 mg / kg body weight of the subject being treated / day, from about 0.01 to about 10 mg / kg body weight / day, about 0.01 mg to about 1 mg / kg body weight / day, from about 1 to about to about 1000 mg / kg body weight / day, from about 5 to about 500 mg / kg body weight / day, from about 10 to about 250 mg / kg body weight / day, about 25 to about 150 mg / kg body weight / day, or about 10 mg / kg body weight / day. In an aspect, the concentration of the compounds in the pharmaceutical composition is at least 0.05 mg / ml (e.g., at least about 0.1 mg / ml, at least about 0.2 mg / ml, at least about 0.5 mg / ml, or at least about 1 mg / ml). Methods of Use In an aspect, the invention provides methods of inhibiting kinase activity in a subject, the method comprising administering to the subject a compound or pharmaceutical composition of the invention. As used herein, “inhibiting” does not necessarily mean 100% reduction in activity, but can mean about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% reduction in activity. In some aspects of the invention, the kinase is a PKA, a PKG, a PKC, a STK, a CLK, a DYRK, or LATS. In some aspects of the invention, the kinase is PKA, PKA / DNAJ, cAMP-PKA, PKG1a, PKG1b, PKG2, PfPKG, PKC-θ, PKC-nu, PKC-d, PKC-eta, PKC-g, STK39, CLK1, CLK2, CLK3, CLK4, DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, LATS1, or LATS2. In some aspects of the invention, the kinase is PKA, PKA / DNAJ, or cAMP-PKA.In some aspects of the invention, the kinase is protein kinase A (PKA). In an aspect, PKA is inhibited resulting in therapeutic benefits to the subject. In an aspect, PKA is inhibited resulting in treatment of a cancer. In an aspect, PKA is inhibited resulting in treatment of liver cancer, for example, hepatocellular carcinoma (HCC) and fibrolamellar hepatocellular carcinoma. In an aspect, PKA is inhibited resulting in treatment of diabetic neuropathic pain (Ma, et al., Neuroscience Letters, 750: 135763 (2021)). In some aspects of the invention, the kinase is J-PKAcα. In an aspect, J-PKAcα is inhibited resulting in therapeutic benefits to the subject. In an aspect, J-PKAcα is inhibited resulting in treatment of a cancer. In an aspect, J-PKAcα is inhibited resulting in treatment of liver cancer, for example, hepatocellular carcinoma (HCC) and fibrolamellar hepatocellular carcinoma. In an aspect, J-PKAcα is inhibited resulting in treatment of diabetic neuropathic pain. In some aspects of the invention, the kinase is cyclic adenosine monophosphate- protein kinase A (cAMP-PKA). In an aspect, cAMP-PKA is inhibited resulting in therapeutic benefits to the subject. In an aspect, cAMP-PKA is inhibited resulting in treatment of a cancer. In an aspect, cAMP-PKA is inhibited resulting in treatment of liver cancer, for example, hepatocellular carcinoma (HCC) and fibrolamellar hepatocellular carcinoma. In an aspect, cAMP-PKA is inhibited resulting in treatment of diabetic neuropathic pain. In some aspects of the invention, the kinase inhibited by compounds of aspects of the invention is a protein kinase G (PKG). In an aspect, a PKG is inhibited resulting in therapeutic benefits to the subject. In an aspect, a PKG is inhibited resulting in treatment of a cancer, for example, gastric cancer or colon cancer (Wu, et al., Molecular Medicine Reports, 14: 1849-1856 (2016); Islam, et al., Carcinogenesis, 43(6): 584–593 (2022)). In an aspect, a PKG is inhibited resulting in treatment and / or prevention of an infection, for example, a parasite infection, for example, malaria (i.e., infection caused by a Plasmodium) (Eck, et al., ChemBioChem, 23(7): 1-8 (2022)). In some aspects of the invention, the kinase is PKG1a, PKG1b, PKG2, or PfPKG. In some aspects of the invention, the kinase is PKG1a. In an aspect, PKG1a is inhibited resulting in therapeutic benefits to the subject. In an aspect, PKG1a is inhibited resulting in treatment of a cancer, for example, gastric cancer or colon cancer. In an aspect, PKG1a is inhibited resulting in treatment and / or prevention of an infection, for example, malaria.In some aspects of the invention, the kinase is PKG1b. In an aspect, PKG1b is inhibited resulting in therapeutic benefits to the subject. In an aspect, PKG1b is inhibited resulting in treatment of a cancer, for example, gastric cancer or colon cancer. In an aspect, PKG1b is inhibited resulting in treatment and / or prevention of an infection, for example, malaria.

[0100] In some aspects of the invention, the kinase is PKG2. In an aspect, PKG2 is inhibited resulting in therapeutic benefits to the subject. In an aspect, PKG2 is inhibited resulting in treatment of a cancer, for example, gastric cancer or colon cancer. In an aspect, PKG2 is inhibited resulting in treatment and / or prevention of an infection, for example, malaria.

[0101] In some aspects of the invention, the kinase is PfPKG from Plasmodium falciparum. In an aspect, PfPKG is inhibited resulting in therapeutic benefits to the subject. In an aspect, PfPKG is inhibited resulting in treatment and / or prevention of an infection, for example, malaria.

[0102] In some aspects of the invention, the kinase is a protein kinase C (PKC).

[0103] In some aspects of the invention, a PKC is inhibited resulting in therapeutic benefits to the subject. In an aspect, a PKC is inhibited resulting in treatment of a cancer.

[0104] In some aspects of the invention, the kinase is PKC-θ, PKC-nu, PKC-d, PKC-eta, or PKC-g. In an aspect of the invention, the kinase is PKC-θ. In an aspect of the invention, the kinase is PKC-nu. In an aspect of the invention, the kinase is PKC-d. In an aspect of the invention, the kinase is PKC-eta. In an aspect of the invention, the kinase is PKC-g.

[0105] In some aspects of the invention, the kinase is a serine / threonine kinase (STK). In an aspect, a STK is inhibited resulting in therapeutic benefits to the subject. In an aspect, a STK is inhibited resulting in treatment of a cancer, for example, breast cancer.

[0106] In some aspects of the invention, the kinase is STK39. In an aspect, STK39 is inhibited resulting in therapeutic benefits to the subject. In an aspect, STK39 is inhibited resulting in treatment of a cancer, for example, breast cancer.

[0107] In some aspects of the invention, the kinase inhibited by compounds of aspects of the invention is a dual-specificity tyrosine-regulated kinase (DYRK). In an aspect, a DYRK is inhibited resulting in therapeutic benefits to the subject. In an aspect, a DYRK is inhibited resulting in treatment of a cancer, for example, gastric cancer or colon cancer (Boni, et al., Cancers, 12: 1-26 (2020); Henderson, et al., J. Med. Chem., 64: 11709−11728 (2021)). In an aspect, a DYRK is inhibited resulting in treatment and / or prevention of an infection, forexample, an infection caused by a protozoa (Loaec, et al., Mar. Drugs, 15(316): 1-15 (2017)) or parasite (e.g., Trypanosoma brucei; Cayla, et al., eLife, 1-34 (2020)). In a further aspect, a DYRK is inhibited resulting in treatment and / or prevention of a neurodegenerative disease. In a further aspect, a DYRK is inhibited resulting in treatment and / or prevention of Down Syndrome.

[0108] In some aspects of the invention, the kinase is DYRK1 A, DYRK1B, DYRK2, DYRK3, or DYRK4.

[0109] In some aspects of the invention, the kinase is DYRK1 A. In an aspect, DYRK1 A is inhibited resulting in therapeutic benefits to the subject. In an aspect, DYRK1 A is inhibited resulting in treatment of a cancer, for example, gastric cancer or colon cancer. In an aspect, DYRK1 A is inhibited resulting in treatment and / or prevention of an infection, for example, an infection caused by a protozoa or parasite. In another aspect, DYRK1 A is inhibited resulting in treatment and / prevention of Down Syndrome. In an aspect, DYRK 1 A is inhibited resulting in treatment and / or prevention of an neurodegeneration associated with Down Syndrome. DYRK1A is a gene located on chromosome 21 that is implicated in Down syndrome. Overexpression of DYRK 1 A may contribute to some of the cognitive and developmental deficits often seen in individuals with Down Syndrome.In some aspects of the invention, the kinase is DYRK1B. In an aspect, DYRK1B is inhibited resulting in therapeutic benefits to the subject. In an aspect, DYRK1B is inhibited resulting in treatment of a cancer, for example, gastric cancer or colon cancer. In an aspect, DYRK1B is inhibited resulting in treatment and / or prevention of an infection, for example, an infection caused by a protozoa or parasite.

[0110] In some aspects of the invention, the kinase is DYRK2. In an aspect, DYRK2 is inhibited resulting in therapeutic benefits to the subject. In an aspect, DYRK2 is inhibited resulting in treatment of a cancer, for example, gastric cancer or colon cancer. In an aspect, DYRK2 is inhibited resulting in treatment and / or prevention of an infection, for example, an infection caused by a protozoa or parasite.

[0111] In some aspects of the invention, the kinase is DYRK3. In an aspect, DYRK3 is inhibited resulting in therapeutic benefits to the subject. In an aspect, DYRK3 is inhibited resulting in treatment of a cancer, for example, gastric cancer or colon cancer. In an aspect, DYRK3 is inhibited resulting in treatment and / or prevention of an infection, for example, an infection caused by a protozoa or parasite.

[0112] In some aspects of the invention, the kinase is DYRK4. In an aspect, DYRK4 is inhibited resulting in therapeutic benefits to the subject. In an aspect, DYRK4 is inhibited resulting in treatment of a cancer, for example, gastric cancer or colon cancer. In an aspect, DYRK4 is inhibited resulting in treatment and / or prevention of an infection, for example, an infection caused by a protozoa or parasite.

[0113] In some aspects of the invention, the kinase inhibited by compounds of aspects of the invention is a Cdc2-like kinase (CLK). In an aspect, a CLK is inhibited resulting in therapeutic benefits to the subject. In an aspect, a CLK is inhibited resulting in treatment of cancer. In an aspect, a CLK is inhibited resulting in treatment of gastric cancer (Tam, et al., Cancer Letters, 473: 186–197 (2020)), pancreatic cancer (Chen, et al., J. Hematol. Oncol., 14: 60 (2021)), prostate cancer (Uzor, et al., Scientific Reports, 11: 7963 (2021)), breast cancer (e.g., triple negative breast cancer) (Riggs, et al., J. Med. Chem., 60: 8989-9002 (2017); Yoshida, et al., Cancer Res, 75(7): 1515-1526 (2015)), lung cancer (e.g., non-small cell) (Liu, et al., JBUON, 26(1): 58-64 (2021)), and / or a glioma (Park, et al., Am. J. Cancer Res., 10(11): 3765-3783 (2020)). In an aspect, a CLK is inhibited resulting in treatment or prevention of memory impairments and neurotoxicityinduced by oligomeric Aβ25–35 peptide administration (Naert, et al., European Neuropsychopharmacology, 2170–2182 (2015); Tam, et al., Cancer Letters, 473: 186–197 (2020); Moyano, et al., Int. J. Mol. Sci., 21: 7549 (2020); and Qin, et al., J. Med. Chem., 64: 13191−13211 (2021)). In an aspect, a CLK is inhibited resulting in treatment or prevention of arthritis, e.g., osteoarthritis (e.g., knee osteoarthritis) (Sun, et al., J. Med. Chem., 67: 6, 4603–4623 (2024)). In an aspect, a CLK is inhibited resulting in treatment or prevention of arthritis, e.g., osteoarthritis (e.g., knee osteoarthritis).

[0114] In some aspects of the invention, the compounds of aspects of the invention modulate RNA splicing. It has been reported that CLK inhibition may function as a pre- mRNA splicing modulation-based anti-cancer strategy, e.g., for MYC-driven cancers (Iwai, et al., EMBO Molecular Medicine, 10: e8289 (2018); Duncan, et al., Experimental Cell Research, 241: 300–308 (1998)). Accordingly, in an aspect, a CLK is inhibited resulting in therapeutic mRNA splicing modulation.

[0115] In some aspects of the invention, the kinase is CLK1, CLK2, CLK3, or CLK4.

[0116] In some aspects of the invention, CLK1 is inhibited resulting in therapeutic benefits to the subject. In an aspect, CLK1 is inhibited resulting in treatment of a cancer, forexample, gastric cancer. In an aspect, CLK1 is inhibited resulting in prevention of memory impairments and neurotoxicity induced by oligomeric AP25-35 peptide administration.

[0117] In some aspects of the invention, CLK2 is inhibited resulting in therapeutic benefits to the subject. In an aspect, CLK2 is inhibited resulting in treatment of a cancer, for example, gastric cancer. In an aspect, CLK2 is inhibited resulting in prevention of memory impairments and neurotoxicity induced by oligomeric AP25-35 peptide administration.

[0118] In some aspects of the invention, CLK3 is inhibited resulting in therapeutic benefits to the subject. In an aspect, CLK3 is inhibited resulting in treatment of a cancer, for example, gastric cancer. In an aspect, CLK3 is inhibited resulting in prevention of memory impairments and neurotoxicity induced by oligomeric AP25-35 peptide administration.

[0119] In some aspects of the invention, CLK4 is inhibited resulting in therapeutic benefits to the subject. In an aspect, CLK4 is inhibited resulting in treatment of a cancer, for example, gastric cancer. In an aspect, CLK4 is inhibited resulting in prevention of memory impairments and neurotoxicity induced by oligomeric AP25-35 peptide administration.

[0120] In some aspects of the invention, the kinase inhibited by compounds of aspects of the invention is a LATS (Large Tumor Suppressor Kinase). In an aspect, a LATS is inhibited resulting in therapeutic benefits to the subject. In an aspect, a LATS is inhibited resulting in treatment of a cancer.

[0121] In some aspects of the invention, the kinase inhibited by compounds of aspects of the invention is LATS1 (Large Tumor Suppressor Kinase 1). In an aspect, LATS1 is inhibited resulting in therapeutic benefits to the subject. In an aspect, a LATS1 is inhibited resulting in treatment of a cancer.

[0122] In some aspects of the invention, the kinase inhibited by compounds of aspects of the invention is LATS2 (Large Tumor Suppressor Kinase 2). In an aspect, LATS2 is inhibited resulting in therapeutic benefits to the subject. In an aspect, a LATS2 is inhibited resulting in treatment of a cancer.

[0123] In some aspects, the invention provides methods of suppressing the immune system in a subject, the method comprising administering to the subject a compound, or a pharmaceutically acceptable salt thereof, or pharmaceutical composition of an aspect of the invention.

[0124] In some aspects, the invention provides methods preventing organ rejection in a subject, the method comprising administering to the subject a compound, or apharmaceutically acceptable salt thereof, or pharmaceutical composition of an aspect of the invention.

[0125] In some aspects, the invention provides methods of treating diabetic neuropathic pain in a subject, the method comprising administering to the subject a compound, or a pharmaceutically acceptable salt thereof, or pharmaceutical composition of an aspect of the invention.

[0126] In some aspects, the invention provides methods of treating malaria in a subject, the method comprising administering to the subject a compound, or a pharmaceutically acceptable salt thereof, or pharmaceutical composition of an aspect of the invention.

[0127] In some aspects, the invention provides methods of treating an infection associated with a protozoa in a subject, the method comprising administering to the subject a compound, or a pharmaceutically acceptable salt thereof, or pharmaceutical composition of an aspect of the invention.

[0128] In some aspects, the invention provides methods of treating a neurodegenerative disease, the method comprising administering to the subject a compound, or a pharmaceutically acceptable salt thereof, or pharmaceutical composition of an aspect of the invention. In an aspect, the invention provides methods of treating Down syndrome, the method comprising administering to the subject a compound, or a pharmaceutically acceptable salt thereof, or pharmaceutical composition of an aspect of the invention. In an aspect, the invention provides methods of treating Alzheimer's disease, the method comprising administering to the subject a compound or pharmaceutical composition of an aspect of the invention.

[0129] In some aspects, the invention provides methods of treating cardiac disease, the method comprising administering to the subject a compound, or a pharmaceutically acceptable salt thereof, or pharmaceutical composition of an aspect of the invention. In an aspect, the invention provides methods of treating heart failure, the method comprising administering to the subject a compound, or a pharmaceutically acceptable salt thereof, or pharmaceutical composition of an aspect of the invention.

[0130] In some aspects, the invention provides methods of treating Cushing’s syndrome, the method comprising administering to the subject a compound, or a pharmaceutically acceptable salt thereof, or pharmaceutical composition of an aspect of the invention.

[0131] In some aspects, the invention provides methods of treating McCune-Albright Syndrome, the method comprising administering to the subject a compound, or apharmaceutically acceptable salt thereof, or pharmaceutical composition of an aspect of the invention.

[0132] In some aspects, the invention provides methods of treating Carney complex, the method comprising administering to the subject a compound, or a pharmaceutically acceptable salt thereof, or pharmaceutical composition of an aspect of the invention.

[0133] An aspect of the invention provides compounds and pharmaceutically compositions for use in treating or preventing cancer. Without being bound by a particular theory or mechanism, it is believed that the compounds inhibit kinases.

[0134] The terms “treat” and “prevent” as well as words stemming therefrom, as used herein, do not necessarily imply 100% or complete treatment or prevention. Rather, there are varying degrees of treatment or prevention of which one of ordinary skill in the art recognizes as having a potential benefit or therapeutic effect. In this respect, the methods of an aspect of the invention can provide any amount of any level of treatment or prevention of cancer in a subject (e.g., a mammal). Furthermore, the treatment or prevention provided by the method of an aspect of the invention can include treatment or prevention of one or more conditions or symptoms of the disease, e.g., cancer, being treated or prevented. Also, for purposes herein, “prevention” can encompass delaying the onset of the disease, or a symptom or condition thereof.

[0135] With respect to the methods of aspects of the invention, the cancer can be any cancer, including any of adrenal gland cancer, sarcomas (e.g., synovial sarcoma, osteogenic sarcoma, leiomyosarcoma uteri, angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma, myxoma, rhabdomyoma, fibroma, lipoma, and teratoma), lymphomas (e.g., small lymphocytic lymphoma, Hodgkin lymphoma, and non-Hodgkin lymphoma), hepatocellular carcinoma, glioma, head cancers (e.g., squamous cell carcinoma), neck cancers (e.g., squamous cell carcinoma), acute lymphocytic cancer, leukemias (e.g., hairy cell leukemia, myeloid leukemia (acute and chronic), lymphatic leukemia (acute and chronic), prolymphocytic leukemia (PLL), myelomonocytic leukemia (acute and chronic), and lymphocytic leukemia (acute and chronic)), bone cancer (osteogenic sarcoma, fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing’s sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor, chordoma, osteochondroma (osteocartilaginous exostoses), benign chondroma, chondroblastoma, chondromyxoid fibroma, osteoid osteoma, and giant cell tumors), brain cancer (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma (pinealoma), glioblastoma multiforme,oligodendroglioma, schwannoma, and retinoblastoma), fallopian tube cancer, breast cancer, cancer of the anus, anal canal, or anorectum, cancer of the eye, cancer of the intrahepatic bile duct, cancer of the joints, cancer of the neck, gallbladder, or pleura, cancer of the nose, nasal cavity, or middle ear, cancer of the oral cavity, cancer of the vulva (e.g., squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, and fibrosarcoma), myeloproliferative disorders (e.g., chronic myeloid cancer), colon cancers (e.g., colon carcinoma), esophageal cancer (e.g., squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, and lymphoma), cervical cancer (cervical carcinoma and pre-invasive cervical dysplasia), gastric cancer, gastrointestinal carcinoid tumor, hypopharynx cancer, larynx cancer, liver cancers (e.g., hepatocellular carcinoma, fibrolamellar carcinoma (FLC), fibrolamellar hepatocellular carcinoma (FL-HCC), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, and hemangioma), lung cancers (e.g., bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, and adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, chondromatous hamartoma, small cell lung cancer, non-small cell lung cancer, and lung adenocarcinoma), malignant mesothelioma, skin cancer (e.g., melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi’s sarcoma, nevi, dysplastic nevi, lipoma, angioma, dermatofibroma, and keloids), multiple myeloma, nasopharynx cancer, ovarian cancer (e.g., ovarian carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, endometrioid carcinoma, and clear cell adenocarcinoma), granulosa-theca cell tumors, Sertoli-Leydig cell tumors, dysgerminoma, and malignant teratoma), pancreatic cancer (e.g., ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors, and VIPoma), peritoneum, omentum, mesentery cancer, pharynx cancer, prostate cancer (e.g., adenocarcinoma and sarcoma), rectal cancer, kidney cancer (e.g., adenocarcinoma, Wilms tumor (nephroblastoma), and renal cell carcinoma), small intestine cancer (adenocarcinoma, lymphoma, carcinoid tumors, Kaposi’s sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, and fibroma), soft tissue cancer, stomach cancer (e.g., carcinoma, lymphoma, and leiomyosarcoma), testicular cancer (e.g., seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, Leydig cell tumor, fibroma, fibroadenoma, adenomatoid tumors, and lipoma), cancer of the uterus (e.g., endometrial carcinoma), thyroid cancer, and urothelial cancers (e.g., squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma, ureter cancer, and urinary bladder cancer). In an aspect of the invention, the cancer is hepatocellular carcinoma. In an aspect of the invention, the cancer is fibrolamellar carcinoma (FLC). In an aspect of the invention, thecancer is fibrolamellar hepatocellular carcinoma (FL-HCC). In an aspect of the invention, the cancer is liver cancer. In an aspect of the invention, the cancer is breast cancer (e.g., triple negative breast cancer). In an aspect of the invention, the cancer is gastric cancer. In an aspect of the invention, the cancer is colon cancer. In an aspect of the invention, the cancer is prostate canceer. In an aspect of the invention, the cancer in pancreatic cancer. In an aspect of the invention, the cancer is a glioma.

[0136] In certain aspects of the invention, the compounds of aspects of the invention, or pharmaceutically acceptable salts thereof, can be co-administered with an anti-cancer agent (e.g., a chemotherapeutic agent) and / or radiation therapy. In an aspect, the compounds of aspects of the invention, or pharmaceutically acceptable salts thereof, are administered in an amount that is effective to sensitize the cancer cells to one or more therapeutic regimens (e.g., chemotherapy or radiation therapy). The terms “co-administered” or “co-administration” refer to simultaneous or sequential administration. The compounds of aspects of the invention, or pharmaceutically acceptable salts thereof, can be administered before, concurrently with, or after administration of another anti-cancer agent (e.g., a chemotherapeutic agent).

[0137] One or more than one, e.g., two, three, or more anti-cancer agents can be administered. In this regard, the present invention is directed a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a combination of the compounds of aspects of the invention, or pharmaceutically acceptable salts thereof, and at least one anticancer agent (e.g., chemotherapeutic agent).

[0138] Examples of anti-cancer agents include platinum compounds (e.g., cisplatin, carboplatin, oxaliplatin), alkylating agents (e.g., cyclophosphamide, ifosfamide, chlorambucil, nitrogen mustard, thiotepa, melphalan, busulfan, procarbazine, streptozocin, temozolomide, dacarbazine, bendamustine), antitumor antibiotics (e.g., daunorubicin, doxorubicin, idarubicin, epirubicin, mitoxantrone, bleomycin, mitomycin C, plicamycin, dactinomycin), taxanes (e.g., paclitaxel and docetaxel), antimetabolites (e.g., 5-fluorouracil, cytarabine, pemetrexed, thioguanine, floxuridine, capecitabine, and methotrexate), nucleoside analogues (e.g., fludarabine, clofarabine, cladribine, pentostatin, nelarabine), topoisomerase inhibitors (e.g., topotecan and irinotecan), hypomethylating agents (e.g., azacitidine and decitabine), proteosome inhibitors (e.g., bortezomib), epipodophyllotoxins (e.g., etoposide and teniposide), DNA synthesis inhibitors (e.g., hydroxyurea), vinca alkaloids (e.g., vincristine, vindesine, vinorelbine, and vinblastine), tyrosine kinase inhibitors (e.g., imatinib,dasatinib, nilotinib, sorafenib, sunitinib), monoclonal antibodies (e.g., rituximab, cetuximab, panitumumab, tositumomab, trastuzumab, alemtuzumab, gemtuzumab ozogamicin, bevacizumab), nitrosoureas (e.g., carmustine, fotemustine, and lomustine), enzymes (e.g., L- Asparaginase), biological agents (e.g., interferons and interleukins), hexamethylmelamine, mitotane, angiogenesis inhibitors (e.g., thalidomide, lenalidomide), steroids (e.g., prednisone, dexamethasone, and prednisolone), hormonal agents (e.g., tamoxifen, raloxifene, leuprolide, bicalutamide, granisetron, flutamide), aromatase inhibitors (e.g., letrozole and anastrozole), arsenic trioxide, tretinoin, nonselective cyclooxygenase inhibitors (e.g., nonsteroidal antiinflammatory agents, salicylates, aspirin, piroxicam, ibuprofen, indomethacin, naprosyn, diclofenac, tolmetin, ketoprofen, nabumetone, oxaprozin), selective cyclooxygenase-2 (COX- 2) inhibitors, cellular immunotherapy (e.g., chimeric antigen receptor T cell therapy, tumorinfiltrating lymphocyte therapy), or any combination thereof. In some aspects, the anticancer agent is cisplatin, cytarabine, methotrexate, doxorubicin, or a combination thereof.

[0139] In certain aspects of the invention, the compounds of aspects of the invention, or pharmaceutically acceptable salts thereof, can be attached to targeting molecules. Such targeting molecules include antibodies (for ADCs) and small molecules that target other regions of kinases to afford more selectivity (i.e., a second molecule that binds the DNAJ domain of the J-PKACα fusion protein).

[0140] In certain aspects of the invention, the compounds of aspects of the invention, or pharmaceutically acceptable salts thereof, can be attached to an E3 ligase binding molecule to make a proteolysis-targeting chimeras (PROTAC).

[0141] In some embodiments, the subject is a mammal. As used herein, the term “mammal” refers to any mammal, including, but not limited to, mammals of the order Rodentia, including mice and hamsters, mammals of the order Logomorpha, including rabbits, mammals from the order Carnivora, including Felines (cats) and Canines (dogs), mammals from the order Artiodactyla, including Bovines (cows) and Swines (pigs), mammals from the order Perssodactyla, including Equines (horses), mammals of the order Primates, Ceboids, or Simoids (monkeys), and mammals of the order Anthropoids (humans and apes). An especially preferred mammal is the human.Examples of Non-Limiting Aspects of the Disclosure

[0142] Aspects, including embodiments, of the present subject matter described herein may be beneficial alone or in combination, with one or more other aspects or embodiments. Without limiting the foregoing description, certain non-limiting aspects of the disclosurenumbered 1-29 are provided below. As will be apparent to those of skill in the art upon reading this disclosure, each of the individually numbered aspects may be used or combined with any of the preceding or following individually numbered aspects. This is intended to provide support for all such combinations of aspects and is not limited to combinations of aspects explicitly provided below:

[0143] (1) A compound of formula (I)whereinis a single or double bond, X1and X2are each independently CH, CR4, or N; X3is S, S=O, or S(=O)2; R1is H or –NR2R3; R2is H, C1-C3alkyl, or an aryl; R3is H, C1-C3alkyl, or an aryl; R4is C1-C3alkyl; and E is C3-C8heterocycloalkyl, C8-C10bicycloalkyl, C4-C10biheterocycloalkyl, aryl, or heteroaryl, each of which is optionally substituted with one or more substituents selected from C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, trifluoromethyl, C1-C6alkoxy, arylated C1-C6alkoxy, trifluoromethylated C1-C6alkoxy, -C(O)OH, -C(O)O-(C1-C6alkyl), -C(O)-(C1-C6alkyl), -(C1-C3alkyl)-C(O)OH, -(C1-C3alkyl)-C(O)O-(C1-C6alkyl), -(C1-C3alkyl)-C(O)-(C1- C6alkyl), -NH-C(O)OH, -NH-C(O)O-(C1-C6alkyl), -NH-C(O)-(C1-C6alkyl), -(C1-C3alkyl)- NH-C(O)OH, -(C1-C3alkyl)-NH-C(O)O-(C1-C6alkyl), -(C1-C3alkyl)-NH-C(O)-(C1-C6alkyl), -(C1-C6alkyl)-O-(C1-C6alkyl), -O-(C1-C6alkyl)-O-(C1-C6alkyl), -(C1-C6alkyl)-OH, - (C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-COOH, -(C1-C6alkyl)-C(O)O-(C1-C6alkyl), -(C1-C6alkyl)-(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-NH2, -(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)- (C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)-heterocycloalkyl, -(C1-C6alkyl)-heterocycloalkyl- (C1-C6alkyl), -(C1-C6alkyl)-C(O)-heterocycloalkyl, -(C1-C6alkyl)-C(O)-heterocycloalkyl- (C1-C6alkyl), halo, nitro, hydroxy, amino, C1-C6alkylamino, di-C1-C6alkyl-amino; -NH- aryl, -NH-heterocycloalkyl, -NH-heterocycloalkyl-(C1-C6alkyl), -C1-C6haloalkyl, C3-C8cycloalkyl, C3-C8heterocycloalkyl, fused C3-C8cycloalkyl, spiro C3-C8cycloalkyl, fused C3- C8heterocycloalkyl, spiro C3-C8heterocycloalkyl, aryl, heteroaryl, fused aryl, fused heteroaryl, -CN, -(C1-C3alkyl)-CN, carbonyl, amino acid, amino acid methyl ester, and a combination thereof, or a pharmaceutically acceptable salt thereof.

[0144] (2) The compound of aspect 1, wherein E is a carbon bound C3-C8heterocycloalkyl, aryl, or heteroaryl, each of which is optionally substituted with one or more substituents selected from C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, trifluoromethyl, C1-C6alkoxy, arylated C1-C6alkoxy, trifluoromethylated C1-C6alkoxy, -(C1-C6alkyl)-OH, -(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-COOH, -(C1-C6alkyl)- NH2, -(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)-(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl) heterocycloalkyl, halo, nitro, hydroxy, amino, C1-C6alkylamino, di-C1-C6alkyl-amino; -NH-aryl, C1-C6haloalkyl, C3-C8cycloalkyl, C3-C8heterocycloalkyl, fused C3- C8cycloalkyl, fused C3-C8heterocycloalkyl, aryl, heteroaryl, fused aryl, fused heteroaryl, - CN, -(C1-C3alkyl)-CN, carbonyl, and a combination thereof, or a pharmaceutically acceptable salt thereof.

[0145] (3) The compound of aspect 1, wherein E is a carbon bound C3-C8heteroaryl, which is optionally substituted with one or more substituents selected from C1-C6alkyl, C2- C6alkenyl, C2-C6alkynyl, trifluoromethyl, C1-C6alkoxy, arylated C1-C6alkoxy, trifluoromethylated C1-C6alkoxy, -(C1-C6alkyl)-OH, -(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-COOH, -(C1-C6alkyl)-NH2, -(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)-(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl) heterocycloalkyl, halo, nitro, hydroxy, amino, C1-C6alkylamino, di-C1-C6alkyl-amino; -NH-aryl, C1-C6haloalkyl, C3-C8cycloalkyl, C3-C8heterocycloalkyl, fused C3-C8cycloalkyl, fused C3-C8heterocycloalkyl, aryl, heteroaryl, fused aryl, fused heteroaryl, -CN, -(C1-C3alkyl)-CN, carbonyl, and a combination thereof, or a pharmaceutically acceptable salt thereof.

[0146] (4) The compound of any one of aspects 1-3, whereinis a double bond, or a pharmaceutically acceptable salt thereof.

[0147] (5) The compound of any one of aspects 1-4, wherein R1is H, or a pharmaceutically acceptable salt thereof.

[0148] (6) The compound of any one of aspects 1-5, wherein the compound of formula (I) is of formula (Ia): ,or a pharmaceutically acceptable salt thereof.

[0149] (7) The compound of any one of aspects 1-6, wherein the compound of formula (I) is of formula (Ib), (Ic), (Id), (Ie), or (1f):37or a pharmaceutically acceptable salt thereof.

[0150] (8) The compound of any one of aspects 1-7, wherein E is selected from:,each of which is optionally substituted with one or more substituents selected from C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, trifluoromethyl, C1-C6alkoxy, arylated C1-C6alkoxy, trifluoromethylated C1-C6alkoxy, -C(O)OH, -C(O)O-(C1-C6alkyl), -C(O)-(C1-C6alkyl), - (C1-C3alkyl)-C(O)OH, -(C1-C3alkyl)-C(O)O-(C1-C6alkyl), -(C1-C3alkyl)-C(O)-(C1-C6alkyl), -NH-C(O)OH, -NH-C(O)O-(C1-C6alkyl), -NH-C(O)-(C1-C6alkyl), -(C1-C3alkyl)- NH-C(O)OH, -(C1-C3alkyl)-NH-C(O)O-(C1-C6alkyl), -(C1-C3alkyl)-NH-C(O)-(C1-C6alkyl), -(C1-C6alkyl)-O-(C1-C6alkyl), -O-(C1-C6alkyl)-O-(C1-C6alkyl), -(C1-C6alkyl)-OH, - (C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-COOH, -(C1-C6alkyl)-C(O)O-(C1-C6alkyl), -(C1-C6alkyl)-(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-NH2, -(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)- (C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)-heterocycloalkyl, -(C1-C6alkyl)-heterocycloalkyl- (C1-C6alkyl), -(C1-C6alkyl)-C(O)-heterocycloalkyl, -(C1-C6alkyl)-C(O)-heterocycloalkyl- (C1-C6alkyl), halo, nitro, hydroxy, amino, C1-C6alkylamino, di-C1-C6alkyl-amino; -NH- aryl, -NH-heterocycloalkyl, -NH-heterocycloalkyl-(C1-C6alkyl), -C1-C6haloalkyl, C3-C8cycloalkyl, C3-C8heterocycloalkyl, fused C3-C8cycloalkyl, spiro C3-C8cycloalkyl, fused C3- C8heterocycloalkyl, spiro C3-C8heterocycloalkyl, aryl, heteroaryl, fused aryl, fused heteroaryl, -CN, -(C1-C3alkyl)-CN, carbonyl, amino acid, amino acid methyl ester, and a combination thereof, or a pharmaceutically acceptable salt thereof.

[0151] (9) The compound of aspect 1, wherein the compound of formula (I) isor a pharmaceutically acceptable salt thereof.

[0152] (10) A pharmaceutical composition comprising a compound of any one of aspects 1-9, or a pharmaceutically acceptable salt thereof, and a pharmaceutical carrier.

[0153] (11) A method of inhibiting kinase activity in a subject, the method comprising administering to the subject a compound of any one of aspects 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of aspect 10.

[0154] (12) The method of aspect 11, wherein the kinase is PKA, PKA / DNAJ, cAMP- PKA, PKG1a, PKG1b, PKG2, PfPKG, PKC-θ, PKC-nu, PKC-d, PKC-eta, PKC-g, STK39, CLK1, CLK2, CLK3, CLK4, DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, LATS1, or LATS2.

[0155] (13) The method of aspect 11, wherein the kinase is PKA, PKA / DNAJ, or cAMP- PKA.

[0156] (14) The method of aspect 11, wherein the kinase is PKG1a, PKG1b, PKG2, or PfPKG.

[0157] (15) The method of aspect 11, wherein the kinase is DYRK1A, DYRK1B, DYRK2, DYRK3, or DYRK4.

[0158] (16) The method of aspect 11, wherein the kinase is CLK1, CLK2, CLK3, or CLK4.

[0159] (17) A method of suppressing the immune system in a subject, the method comprising administering to the subject a compound of any one of aspects 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of aspect 10 to the subject.

[0160] (18) A method of preventing organ rejection in a subject, the method comprising administering to the subject a compound of any one of aspects 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of aspect 10 to the subject.

[0161] (19) A method of treating cancer in a subject, the method comprising administering to the subject a compound of any one of aspects 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of aspect 10 to the subject.

[0162] (20) The method of aspect 19, wherein the cancer is fibrolamellar carcinoma(FLC).

[0163] (21) The method of aspect 19, wherein the cancer is fibrolamellar hepatocellular carcinoma (FL-HCC).

[0164] (22) The method of aspect 19, wherein the cancer is gastric cancer, colon cancer, pancreatic cancer, prostate cancer, breast cancer, lung cancer, or a glioma.

[0165] (23) A method of modulating mRNA splicing in a subject, the method comprising administering to the subject a compound of any one of aspects 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of aspect 10 to the subject.

[0166] (24) A method of treating diabetic neuropathic pain in a subject, the method comprising administering to the subject a compound of any one of aspects 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of aspect 10 to the subject.

[0167] (25) A method of treating malaria in a subject, the method comprising administering to the subject a compound of any one of aspects 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of aspect 10 to the subject.

[0168] (26) A method of treating an infection associated with a protozoa in a subject, the method comprising administering to the subject a compound of any one of aspects 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of aspect 10 to the subject.

[0169] (27) A method of treating neurodegenerative disease in a subject, the method comprising administering to the subject a compound of any one of aspects 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of aspect 10 to the subject.

[0170] (28) A method of treating Down Syndrome in a subject, the method comprising administering to the subject a compound of any one of aspects 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of aspect 10 to the subject.

[0171] (29) The method of any one of aspects 11-28, wherein the subject is human.EXAMPLES

[0172] All solvents were of LC-MS grade or better.

[0173] NMR data were obtained on a Bruker Avance III NMR spectrometer equipped with a 3 mm cryogenic probe (600 MHz for1H, 150 MHz for13C). EXAMPLE 1

[0174] This example provides an exemplary synthesis for Acid Scaffolds Compounds A- E and Bromo Scaffolds A-C and E for use in Example 2.

[0175] General Procedure for Synthesis of Acid Scaffold-A of Example 1:

[0176] Acid Scaffold-A was synthesized from the commercially available H-1 and H-2. H-1 was reacted with H-2 in presence of Xantphos Pd G3 and Cs2CO3to afford H-3, which was reacted with KOH to afford Acid Scaffold-A, as shown in Scheme 1. Scheme 1: Preparation of Acid Scaffold-A of Example 1

[0177] Preparation of Ethyl 4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3,4-dihydro-2H-1,4- thiazine-6-carboxylate (H-3): To a mixture of ethyl 3,4-dihydro-2H-1,4-thiazine-6- carboxylate (H-1, 1 equiv., 100 mg) and 4-bromo-7H-pyrrolo[2,3-d]pyrimidine (H-2, 1.1 equiv., 125 mg) was added Xantphos Pd G3 (10 mol%, 55 mg), Cs2CO3(3 equiv., 560 mg), and DMF (5 mL). The reaction vial was filled with N2and capped tightly. The reaction mixture was then stirred vigorously at 110 °C overnight before it was dried under vacuum. The resulting residue was re-dissolved in DMSO followed by preparative HPLC purification using a Gemini 5 μm NX-C18 column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10% to 100% MeCN in 0.1% TFA) to yield H-3 (109 mg, 65% yield) as a pale white solid.1H NMR (600 MHz, DMSO-d6): δ 12.21 (s, 1H), 8.84 (s, 1H), 8.44 (s, 1H), 7.49 (s, 1H), 6.67 (s, 1H), 4.35 (br s, 2H), 4.20 (br s, 2H), 3.13 (br s, 2H), 1.24 (br s, 3H);13C NMR (150 MHz, DMSO-d6): δ 164.7, 152.8, 152.6, 149.9, 133.3, 125.0, 104.5, 100.7, 100.1, 60.4, 45.3, 23.9, 14.3; HRESIMS m / z 291.0910, [M+H]+(calcd for C13H15N4O2S, 291.0916).

[0178] Preparation of 4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine- 6-carboxylic acid (Acid Scaffold-A): H-3 (109 mg) was reacted with KOH (3 equiv., 62 mg) in MeOH (5 mL) and H2O (5 mL) and stirred at 95 °C for 2 h. The reaction solution was acidified with 2M HCl (1.4 mL) and then dried under vacuum. The crude product was then washed and desalted with H2O (1 mL x 3 times) to yield Acid Scaffold-A (80 mg, 81% yield) as a pale white solid.1H NMR (600 MHz, DMSO-d6): δ 12.18 (s, 1H), 8.79 (s, 1H), 8.43 (s, 1H), 7.47 (s, 1H), 6.65 (s, 1H), 4.33 (br s, 2H), 3.12 (br s, 2H);13C NMR (150 MHz, DMSO- d6): δ 166.2, 152.7, 152.6, 149.9, 132.9, 124.8, 104.4, 101.7, 100.1, 44.9, 24.0; HRESIMS m / z 263.0608, [M+H]+(calcd for C11H11N4O2S, 263.0603).

[0179] General Procedure for Synthesis of Acid Scaffold-B of Example 1:

[0180] Acid Scaffold-B was synthesized from the commercially available H-1 and H-4. H-1 was reacted with H-4 in presence of Xantphos Pd G3 and Cs2CO3to afford H-5 which was reacted with KOH to afford Acid Scaffold-B, as shown in Scheme 2. Scheme 2: Preparation of Acid Scaffold-B of Example 1

[0181] Preparation of Ethyl 4-(7H-purin-6-yl)-3,4-dihydro-2H-1,4-thiazine-6- carboxylate (H-5): To a mixture of ethyl 3,4-dihydro-2H-1,4-thiazine-6-carboxylate (H-1, 1 equiv., 100 mg) and 6-bromopurine (H-4, 1.5 equiv., 170 mg) was added Xantphos Pd G3 (10 mol%, 55 mg), Cs2CO3(3 equiv., 560 mg), and DMF (5 mL). The reaction vial was filled with N2and capped tightly. The reaction mixture was then stirred vigorously at 110 °C overnight before it was dried under vacuum. The resulting residue was re-dissolved in DMSO followed by preparative HPLC purification using a Gemini 5 μm NX-C18 column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10% to 100% MeCN in 0.1% TFA) to yield H-5 (61 mg, 36% yield) as a pale white solid.1H NMR (600 MHz, DMSO-d6): δ 13.56 (s, 1H), 9.77 (s, 1H), 8.52 (s, 1H), 8.44 (s, 1H), 4.55 (br s, 2H), 4.20 (q, J = 7.1 Hz, 2H), 3.16 (m, 2H), 1.24 (t, J = 7.1 Hz, 3H);13C NMR (150 MHz, DMSO-d6): δ 164.7, 153.0, 151.3, 149.5, 141.7, 134.6, 120.4, 102.1, 60.5, 43.5, 23.8, 14.4; HRESIMS m / z 292.0865, [M+H]+(calcd for C12H14N5O2S, 292.0863).

[0182] Preparation of 4-(7H-purin-6-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid (Acid Scaffold-B): H-5 (61 mg) was dissolved in 2 M NaOH (3.5 mL) and THF (3.5 mL) and stirred at room temperature overnight. The reaction solution was acidified with 2 M HCl (5 mL) and then dried under vacuum. The crude product was then washed and desalted with H2O (1 mL x 3 times) to yield Acid Scaffold-B (49 mg, 90% yield) as a pale orange solid.1H NMR (600 MHz, DMSO-d6): δ 13.53 (s, 1H), 12.56 (brs, 1H), 9.78 (s, 1H), 8.50 (s, 1H), 8.43 (s, 1H), 4.51 (br s, 2H), 3.13 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 166.3, 152.9, 151.3, 149.5, 141.6, 134.3, 120.2, 103.2, 43.1, 23.7; HRESIMS m / z 264.0554, [M+H]+(calcd for C10H10N5O2S, 264.0550).

[0183] General Procedure for Synthesis of Acid Scaffold-C of Example 1:

[0184] Acid Scaffold-C was synthesized from the commercially available H-1 and H-6. H-1 was reacted with H-6 in presence of Xantphos Pd G3 and Cs2CO3to afford H-7 which was reacted with KOH to afford Acid Scaffold-C, as shown in Scheme 3. Scheme 3: Preparation of Acid Scaffold-C of Example 1

[0185] Preparation of Ethyl 4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H-1,4- thiazine-6-carboxylate (H-7): To a mixture of ethyl 3,4-dihydro-2H-1,4-thiazine-6- carboxylate (H-1, 1 equiv., 100 mg) and 4-bromo-7-azaindole (H-6, 1 equiv., 95 mg) was added Xantphos Pd G3 (10 mol%, 55 mg), Cs2CO3(3 equiv., 560 mg), and DMF (5 mL). The reaction vial was filled with N2and capped tightly. The reaction mixture was then stirred vigorously at 110 °C overnight before it was dried under vacuum. The residue was re- dissolved in DMSO followed by preparative HPLC purification using a Gemini 5 μm NX- C18 column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10% to 100% MeCN in 0.1% TFA) to yield H-7 (78 mg, 47% yield) as a pale white solid.1H NMR (600 MHz, DMSO-d6): δ 12.42 (s, 1H), 8.25 (d, J = 6.0 Hz, 1H), 8.21 (s, 1H), 7.59 (d, J = 3.5 Hz,1H), 6.97 (d, J = 6.0 Hz,1H), 6.69 (d, J = 3.5 Hz,1H), 4.18 (q, J = 7.1 Hz, 2H), 4.17 (m, 2H), 3.18 (m, 2H), 1.29 (t, J = 7.1 Hz, 3H);13C NMR (150 MHz, DMSO-d6): δ 164.5, 147.7, 145.1, 139.2, 135.1, 126.3, 111.6, 104.5, 100.8, 100.1, 60.5, 47.1, 23.9, 14.3; HRESIMS m / z 290.0977, [M+H]+(calcd for C14H16N3O2S, 290.0963).

[0186] Preparation of 4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6- carboxylic acid (Acid Scaffold-C): H-7 (78 mg) was reacted with KOH (3 equiv., 45 mg) in MeOH (3 mL) and H2O (3 mL) and stirred at 95 °C for 2 h. The reaction solution was acidified with 2M HCl (0.99 mL) and then dried under vacuum. The crude product was then washed and desalted with H2O (1 mL x 3 times) to yield Acid Scaffold-C (48 mg, 68% yield) as a pale white solid.1H NMR (600 MHz, DMSO-d6): δ 12.30 (s, 1H), 8.23 (d, J = 6.0 Hz, 1H), 8.18 (s, 1H), 7.57 (d, J = 3.5 Hz,1H), 6.93 (d, J = 6.0 Hz,1H), 6.67 (d, J = 3.5 Hz,1H), 4.15 (m, 2H), 3.17 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 166.0, 147.5, 145.2, 139.4, 134.6, 126.0, 111.3, 104.3, 102.1, 100.1, 46.8, 23.9; HRESIMS m / z 262.0650, [M+H]+(calcd for C12H12N3O2S, 262.0650).

[0187] General Procedure for Synthesis of Acid Scaffold-D of Example 1:

[0188] Acid Scaffold-D was synthesized from readily available H-1 and H-8. H-1 was reacted with H-8 in presence of CuI and Cs2CO3to afford H-9 which was reacted with KOH to afford Acid Scaffold-D, as shown in Scheme 4. Scheme 4: Preparation of Acid Scaffold-D of Example 1

[0189] Preparation of Ethyl 4-(1-methyl-1H-imidazol-5-yl)-3,4-dihydro-2H-1,4-thiazine- 6-carboxylate (H-9): To the mixture of ethyl 3,4-dihydro-2H-1,4-thiazine-6-carboxylate (H- 1, 1 equiv., 100 mg) and 5-iodo-1-methyl-1H-imidazole (H-8, 1.5 equiv., 180 mg) was added CuI (10 mol%, 11 mg), Cs2CO3(3 equiv., 564 mg), N,N'-Dimethylethylenediamine (20 mol%, 12.4 μL), and DMF (5 mL). The reaction vial was filled with N2and capped tightly. The reaction mixture was then stirred vigorously at 110 °C overnight before it was dried under vacuum. The residue was re-dissolved in DMSO followed by preparative HPLC purification using a Gemini 5 μm NX-C18 column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10% to 100% MeCN in 0.1% TFA) to yield H-9 (52 mg, 36% yield) as a pale yellow solid.1H NMR (600 MHz, Methanol-d4): δ 8.85 (s, 1H), 7.58 (br s, 1H), 7.57 (s, 1H), 4.18 (q, J = 7.1 Hz, 2H), 3.85 (m, 2H), 3.81 (s, 3H), 3.11 (m, 2H), 1.26 (t, J = 7.1 Hz, 3H);13C NMR (150 MHz, Methanol-d4): δ 166.9, 138.9, 138.8, 135.4, 115.2,100.3, 61.9, 51.4, 33.3, 24.8, 14.6; HRESIMS m / z 254.0966, [M+H]+(calcd for C11H16N3O2S, 254.0963)

[0190] Preparation of 4-(1-methyl-1H-imidazol-5-yl)-3,4-dihydro-2H-1,4-thiazine-6- carboxylic acid (Acid Scaffold-D): H-9 (52 mg) was reacted with KOH (3 equiv., 29 mg) in MeOH (1 mL) and H2O (1 mL) and stirred at 95 °C for 2 h. The reaction solution was acidified with 2M HCl (0.26 mL) and then dried under vacuum. The crude product was purified by preparative HPLC using a Gemini 5 μm NX-C18 column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10% to 100% MeCN in 0.1% TFA) to yield Acid Scaffold-D (28 mg, 60% yield) as a pale white solid.1H NMR (600 MHz, Methanol-d4): δ 8.83 (s, 1H), 7.57 (s, 1H), 7.56 (br s, 1H), 3.85 (m, 2H), 3.80 (s, 3H), 3.11 (m, 2H);13C NMR (150 MHz, Methanol-d4): δ 168.6, 139.1, 138.9, 135.3, 115.1, 100.8, 51.3, 33.3, 24.8; HRESIMS m / z 226.0645, [M+H]+(calcd for C9H12N3O2S, 226.0650).

[0191] General Procedure for Synthesis of Acid Scaffold-E of Example 1:

[0192] Acid Scaffold-E was synthesized from readily available H-1 and H-10. H-1 was reacted with H-10 in presence of CuI and Cs2CO3, and the resulting product was subsequently reacted with KOH to afford Acid Scaffold-E, as shown in Scheme 5. Scheme 5: Preparation of Acid Scaffold-E of Example 1

[0193] To a mixture of ethyl 3,4-dihydro-2H-1,4-thiazine-6-carboxylate (H-1, 1 equiv., 100 mg) and 4-chloropyrimidine derivatives (H-10, 1.5 equiv.) was added CuI (10 mol%, 11 mg), Cs2CO3(3 equiv., 564 mg), N,N'-Dimethylethylenediamine (20 mol%, 12.4 μL), and DMF (5 mL). The reaction vial was filled with N2and capped tightly. The reaction mixture was then stirred vigorously at 110 °C overnight before it was dried down under vacuum. The residue was reacted with KOH (3 equiv., 97 mg) in MeOH (5 mL) and H2O (1 mL) and stirred at 95 °C for 2 h. The reaction solution was acidified with 2M HCl (0.85 mL) and then dried down under vacuum. The crude product was purified by preparative HPLC using a Gemini 5 μm NX-C18 column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10% to 100% MeCN in 0.1% TFA) to yield Acid Scaffold-E.

[0194] General Procedure for Synthesis of Bromo Scaffold-A of Example 1:

[0195] Bromo Scaffold-A was synthesized from Acid Scaffold-A, as shown in Scheme 6. Scheme 6: Preparation of Bromo Scaffold-A of Example 1

[0196] Preparation of 6-Bromo-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3,4-dihydro-2H- 1,4-thiazine (Bromo Scaffold-A): A reaction vial containing a DMF (8 mL) solution of Acid scaffold-A (1 equiv., 40 mg) and K2CO3(2 equiv., 40 mg) was filled with N2and cooled down in an ice bath. A 40 mg / mL MeCN solution of N-bromosuccinimide (1.2 equiv., 800 μL) was added dropwise into the reaction vial, and the resulting reaction mixture was stirred at 0 °C for 2 h and then warmed up to room temperature with continued stirring overnight. The reaction mixture was dried under vacuum. The resulting residue was re-dissolved in DMSO followed by preparative HPLC purification using a Gemini 5 μm NX-C18 column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10-100% MeCN in 0.1% TFA) to yield Bromo scaffold-A (13 mg, 29% yield) as a pale orange solid.1H NMR (600 MHz, DMSO-d6): δ 12.12 (s, 1H), 8.32 (s, 1H), 7.96 (s, 1H), 7.39 (br s, 1H), 6.64 (br s, 1H), 4.37 (m, 2H), 3.32 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 152.1, 151.8, 149.6, 125.0, 123.9, 103.4, 100.8, 43.8, 28.7; ESIMS m / z 296.98, [M+H]+.

[0197] General Procedure for Synthesis of Bromo Scaffold-B of Example 1:

[0198] Bromo Scaffold-B was synthesized from Acid Scaffold-B, as shown in Scheme 7. Scheme 7: Preparation of Bromo Scaffold-B of Example 1

[0199] Preparation of 6-bromo-4-(7H-purin-6-yl)-3,4-dihydro-2H-1,4-thiazine (Bromo Scaffold-B): A reaction vial containing a DMF (5 mL) solution of Acid scaffold-B (1 equiv., 49 mg) with 4Å molecular sieves was filled with N2and cooled down in an ice bath. Et3N (1 equiv., 49 µL) and a 10 mg / mL MeCN solution of N-bromosuccinimide (1.2 equiv., 3.8 mL) was added dropwise into the reaction vial, and the resulting reaction mixture was stirred at 0 °C for 2 h and then warmed up to room temperature with continued stirring overnight. The reaction mixture was dried under vacuum. The resulting residue was re-dissolved in DMSO followed by preparative HPLC purification using a Gemini 5 μm NX-C18 column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10-100% MeCN in 0.1% TFA) to yield Bromo scaffold-B (25 mg, 45% yield) as a pale orange solid.1H NMR (600 MHz, DMSO-d6): δ 13.36 (s, 1H), 8.98 (s, 1H), 8.37 (s, 1H), 8.31 (s, 1H), 4.56 (br s, 2H), 3.32 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 152.3, 151.3, 149.0, 140.4, 125.7, 119.3, 91.1, 41.7, 28.2; HRESIMS m / z 297.9757, [M+H]+(calcd for C9H9BrN5S, 297.9757).

[0200] General Procedure for Synthesis of Bromo Scaffold-C of Example 1:

[0201] Bromo Scaffold-C was synthesized from Acid Scaffold-C, as shown in Scheme 8. Scheme 8: Preparation of Bromo Scaffold-C of Example 1

[0202] Preparation of 6-Bromo-4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H-1,4- thiazine (Bromo Scaffold-C): A reaction vial containing a DMF (6 mL) solution of Acid scaffold-C (1 equiv., 30 mg)) was filled with N2and cooled down in an ice bath. Et3N (1.2equiv., 36 μL) and a 40 mg / mL MeCN solution of N-bromosuccinimide (1 equiv., 510 μL) was added dropwise into the reaction vial and the resulting reaction mixture was stirred at 0 °C for 2 h and then warmed up to room temperature with continued stirring overnight. The reaction mixture was dried under vacuum. The resulting residue was re-dissolved in DMSO followed by preparative HPLC purification using a Gemini 5 μm NX-C18 column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10-100% MeCN in 0.1% TFA) to yield Bromo scaffold-C (17 mg, 29% yield) as a pale orange solid.1H NMR (600 MHz, DMSO-d6): δ 12.73 (s, 1H), 8.20 (d, J = 6.8 Hz, 1H), 7.54 (d, J = 4.8 Hz, 1H), 7.47 (s, 1H), 6.96 (d, J = 6.8 Hz, 1H), 6.80 (d, J = 4.8 Hz, 1H), 4.27 (m, 2H), 3.38 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 149.2, 140.4, 135.3, 126.0, 125.4, 110.7, 102.5, 102.3, 96.0, 45.7, 28.9; ESIMS m / z 295.98, [M+H]+.

[0203] General Procedure for Synthesis of Bromo Scaffold-E of Example 1:

[0204] Bromo Scaffold-E was synthesized from Acid Scaffold-E, as shown in Scheme 9. Scheme 9: Preparation of Bromo Scaffold-E of Example 1

[0205] A reaction vial containing a DMF (x / 10 mL) solution of Acid scaffold-E (1 equiv., x mg) was filled with N2and cooled down in an ice bath. Et3N (1 equiv.) and a 40 mg / mL MeCN solution of N-bromosuccinimide (1 equiv.) was added dropwise into the reaction vial and the resulting reaction mixture was stirred at 0 °C overnight. The reaction mixture was dried under vacuum and subjected to preparative HPLC purifications using a Gemini 5 μm NX-C18 column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10% to 100% MeCN in 0.1% TFA) to yield the Bromo scaffold-E. EXAMPLE 2

[0206] This example provides an exemplary synthesis for Compounds A-AA. Compounds A-AA were synthesized from Acid Scaffolds Compounds A-E and Bromo Scaffolds A-C and E of Example 1 using the following procedures.

[0207] General Procedure for Suzuki Coupling with Bromo Scaffolds A-C:

[0208] Suzuki products were prepared from Bromo Scaffold-A, B, or C, as shown in Scheme 10. To a mixture of Bromo Scaffold-A, B, or C (1 equiv.) and Boronic Acid Pinacol (BPin) Esters (4 equiv.) was added Xantphos Pd G3 (20 mol%), Cs2CO3(3 equiv.), and DMF / H2O (2 : 1). The reaction vial was filled with N2and capped tightly. The reaction mixture was then stirred vigorously at 95 °C overnight before it was dried under vacuum. The residue was re-dissolved in DMSO followed by successive preparative HPLC purifications using a Gemini 5 μm NX-C18 column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10% to 100% MeCN in 0.1% TFA) and a Synergi 5 μm Hydro-RP column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10-100% MeCN in 0.1% TFA) to yield the Suzuki product. Scheme 10: General Procedure for Suzuki Coupling with Bromo Scaffolds A-C

[0209] Preparation of 4-(9H-purin-6-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro- 2H-1,4-thiazine (Compound A):4-(9H-purin-6-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H-1,4-thiazine (Compound A) was prepared, using a Suzuki coupling with Bromo Scaffold-B and the appropriate boronic acid (7% yield), as a light yellow solid.1H NMR (600 MHz, DMSO-d6): δ 13.46 (s, 1H), 11.94 (s, 1H), 9.51 (br s, 1H), 8.49 (s, 1H), 8.36 (s, 1H), 8.35 (br s, 1H), 7.54 (d, J = 3.5 Hz, 1H), 7.24 (s, 1H), 7.08 (s, 1H), 4.68 (m, 2H), 3.36 (m, 2H);13C NMR (150MHz, DMSO-d6): δ 152.5, 151.4, 149.8, 147.6, 141.3, 140.8, 139.5, 127.4, 126.4, 120.0, 116.9, 113.6, 109.4, 100.5, 43.0, 25.0; HRESIMS m / z 336.1039, [M+H]+(calcd for C16H14N7S, 336.1031).

[0210] Preparation of 6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(7H-pyrrolo[2,3-d]pyrimidin- 4-yl)-3,4-dihydro-2H-1,4-thiazine (Compound B):6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3,4-dihydro-2H-1,4- thiazine (Compound B) was prepared, using a Suzuki coupling with Bromo Scaffold-A and the appropriate boronic acid (12% yield), as a light yellow solid.1H NMR (600 MHz, DMSO-d6): δ 12.18 (s, 1H), 12.16 (s, 1H), 8.70 (s, 1H), 8.41 (s, 1H), 8.31 (br s, 1H), 7.60 (s, 1H), 7.42 (s, 1H), 7.28 (s, 1H), 6.87 (s, 1H), 6.68 (s, 1H), 4.52 (m, 2H), 3.35 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 152.8, 152.5, 150.0, 145.5, 141.5, 139.7, 127.1, 127.0, 124.3, 117.1, 112.3, 107.9, 104.2, 100.7, 100.5, 45.5, 25.2; HRESIMS m / z 335.1077, [M+H]+(calcd for C17H15N6S, 335.1079).

[0211] Preparation of 4,6-bis(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H-1,4-thiazine (Compound C):4,6-bis(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H-1,4-thiazine (Compound C) was prepared, using a Suzuki coupling with Bromo Scaffold-C and the appropriate boronic acid (12% yield), as a light yellow solid.1H NMR (600 MHz, DMSO-d6): δ 12.51 (s, 1H), 12.00 (s, 1H), 8.24 (br s, 2H), 7.82 (s, 1H), 7.52 (s, 2H), 7.24 (br s, 1H), 7.03 (br s, 1H), 6.76 (s, 1H), 6.74 (s, 1H), 4.37 (m, 2H), 3.40 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 149.0,147.2, 142.2, 141.1, 139.0, 136.8, 126.7, 126.5, 125.4, 116.6, 112.5, 111.9, 111.0, 103.5, 101.5, 99.9, 46.7, 25.3; HRESIMS m / z 334.1128, [M+H]+(calcd for C18H16N5S, 334.1126).

[0212] Synthesis of Compound A Derivatives - General Procedure for Alkylation and Deprotection

[0213] Derivatives of Compound A were prepared from Compound A, as shown in Scheme 11. To a mixture of Compound A (1 equiv.) and N-Boc-protected amino alkyl bromides (3 equiv.) was added K2CO3(3 equiv.) and DMF. The reaction mixture was then stirred vigorously at room temperature overnight before it was dried down under vacuum. The residue was separated by preparative HPLC using a Gemini 5 μm NX-C18 column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10% to 100% MeCN in 0.1% TFA) to afford the crude N-Boc-protected products which were further deprotected in DCM : TFA (2 : 1) followed by HPLC purification using a Synergi 5 μm Hydro-RP column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10-100% MeCN in 0.1% TFA) to yield the deprotected products. Scheme 11: Synthesis of Compound A Derivatives

[0214] Preparation of 2-(6-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4- thiazin-4-yl)-9H-purin-9-yl)ethan-1-amine (TFA salt) (Compound D):2-(6-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4-thiazin-4-yl)-9H-purin-9- yl)ethan-1-amine (TFA salt) (Compound D) was prepared, by combining Compound A and the appropriate N-Boc-protected amino alkyl bromide (15% yield), as a light yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.05 (s, 1H), 9.48 (br s, 1H), 8.54 (s, 1H), 8.38 (s, 1H), 8.29 (br s, 1H), 8.00 (br s, 3H), 7.57 (s, 1H), 7.25 (s, 1H), 7.08 (br s, 1H), 4.69 (br s, 2H), 4.49 (m, 2H), 3.37 (m, 4H);13C NMR (150 MHz, DMSO-d6): δ 152.0, 151.5, 149.9, 147.2, 142.7, 141.0, 139.9, 127.0, 126.6, 120.5, 116.6, 112.6, 110.1, 100.5, 43.3, 51.3, 38.3, 25.0; HRESIMS m / z 379.1459, [M+H]+(calcd for C18H19N8S, 379.1453).

[0215] Preparation of 3-(6-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4- thiazin-4-yl)-9H-purin-9-yl)propan-1-amine (TFA salt) (Compound E):3-(6-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4-thiazin-4-yl)-9H-purin-9- yl)propan-1-amine (TFA salt) (Compound E) was prepared, by combining Compound A and the appropriate N-Boc-protected amino alkyl bromide (8% yield), as a light yellow solid;1H NMR (600 MHz, Methanol-d4): δ 9.88 (s, 1H), 8.58 (s, 1H), 8.35 (br s, 1H), 8.31 (s, 1H), 7.73 (br s, 1H), 7.66 (d, J = 3.6 Hz, 1H), 7.59 (br s, 1H), 4.79 (br s, 2H), 4.44 (m, 2H), 3.39 (m, 2H), 2.99 (m, 2H), 2.27 (m, 2H);13C NMR (150 MHz, Methanol-d4): δ 153.4, 152.8, 151.9, 149.3, 143.8, 141.0, 134.5, 133.2, 129.2, 122.6, 122.0, 114.1, 110.4, 104.8, 45.5, 41.8, 38.0, 29.2, 26.1; HRESIMS m / z 393.1613, [M+H]+(calcd for C19H21N8S, 393.1610).

[0216] Preparation of (S)-4-(9-(piperidin-3-ylmethyl)-9H-purin-6-yl)-6-(1H-pyrrolo[2,3- b]pyridin-4-yl)-3,4-dihydro-2H-1,4-thiazine (TFA salt) (Compound F):(S)-4-(9-(piperidin-3-ylmethyl)-9H-purin-6-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4- dihydro-2H-1,4-thiazine (TFA salt) (Compound F) was prepared, by combining Compound A and the appropriate N-Boc-protected amino alkyl bromide (25% yield), as a light yellow solid;1H NMR (600 MHz, Methanol-d4): δ 9.89 (s, 1H), 8.56 (s, 1H), 8.29 (s, 1H), 8.27 (br s, 1H), 7.70 (br s,1H), 7.67 (d, J = 3.6 Hz,1H), 7.58 (br s, 1H), 4.78 (br s, 2H), 4.31 (d, J = 7.0 Hz, 2H), 3.39 (m, 2H), 3.34 (m, 1H), 3.29 (m, 1H), 2.92 (m, 1H), 2.85 (m, 1H), 2.47 (m, 1H), 1.97 (m, 1H), 1.87 (m, 1H), 1.71 (m, 1H), 1.41 (m, 1H);13C NMR (150 MHz, Methanol-d4): δ 153.5, 152.8, 151.9, 149.7, 144.1, 140.7, 134.0, 133.5, 129.2, 122.6, 122.1, 113.8, 110.1, 104.8, 47.7, 47.3, 45.5, 45.2, 35.9, 27.1, 26.0, 22.8; HRESIMS m / z 433.1925, [M+H]+(calcd for C22H25N8S, 433.1923).

[0217] Preparation of (R)-4-(9-(piperidin-3-ylmethyl)-9H-purin-6-yl)-6-(1H-pyrrolo[2,3- b]pyridin-4-yl)-3,4-dihydro-2H-1,4-thiazine (TFA salt) (Compound G):(R)-4-(9-(piperidin-3-ylmethyl)-9H-purin-6-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4- dihydro-2H-1,4-thiazine (TFA salt) (Compound G) was prepared, by combining Compound A and the appropriate N-Boc-protected amino alkyl bromide (3% yield), as a light yellow solid;1H NMR (600 MHz, Methanol-d4): δ 9.89 (s, 1H), 8.56 (s, 1H), 8.29 (s, 1H), 8.27 (br s, 1H), 7.70 (br s,1H), 7.67 (d, J = 3.6 Hz,1H), 7.58 (br s, 1H), 4.78 (br s, 2H), 4.31 (d, J = 7.0 Hz, 2H), 3.39 (m, 2H), 3.34 (m, 1H), 3.29 (m, 1H), 2.92 (m, 1H), 2.85 (m, 1H), 2.47 (m, 1H),1.97 (m, 1H), 1.87 (m, 1H), 1.71 (m, 1H), 1.41 (m, 1H); ; HRESIMS m / z 433.1926, [M+H]+(calcd for C22H25N8S, 433.1923).

[0218] Preparation of (1S,3S)-3-((6-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H- 1,4-thiazin-4-yl)-9H-purin-9-yl)methyl)cyclohexan-1-amine (TFA salt) (Compound H):(1S,3S)-3-((6-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4-thiazin-4-yl)-9H-purin- 9-yl)methyl)cyclohexan-1-amine (TFA salt) (Compound H) was prepared, by combining Compound A and the appropriate N-Boc-protected amino alkyl bromide (11% yield), as a light yellow solid;1H NMR (600 MHz, Methanol-d4): δ 9.90 (s, 1H), 8.55 (s, 1H), 8.29 (br s, 1H), 8.28 (s, 1H), 7.68 (br s, 1H), 7.66 (d, J = 3.6 Hz, 1H), 7.58 (br s, 1H), 4.77 (br s, 2H), 4.30 (m, 2H), 3.60 (m, 1H), 3.38 (m, 2H), 2.45 (m, 1H), 1.91 (m, 1H), 1.75 (m, 1H), 1.59- 1.78 (m, 5H), 1.36 (m, 1H);13C NMR (150 MHz, Methanol-d4): δ 153.5, 152.8, 151.8, 149.8, 144.2, 140.6, 133.9, 133.6, 129.2, 122.5, 121.8, 113.8, 110.0, 104.9, 48.0, 47.7, 45.5, 34.1, 32.9, 30.3, 28.6, 26.0, 20.0; HRESIMS m / z 447.2089, [M+H]+(calcd for C23H27N8S, 447.2079).

[0219] Preparation of (1S,3R)-3-((6-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H- 1,4-thiazin-4-yl)-9H-purin-9-yl)methyl)cyclopentan-1-amine (TFA salt) (Compound I):(1S,3R)-3-((6-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4-thiazin-4-yl)-9H-purin- 9-yl)methyl)cyclopentan-1-amine(TFA salt) (Compound I) was prepared, by combiningCompound A and the appropriate N-Boc-protected amino alkyl bromide (15% yield), as a light yellow solid;1H NMR (600 MHz, Methanol-d4): δ 9.87 (s, 1H), 8.52 (s, 1H), 8.31 (s, 1H), 8.25 (br s, 1H), 7.65 (d, J = 3.6 Hz, 1H), 7.63 (m, 1H), 7.57 (br s, 1H), 4.74 (br s, 2H), 4.34 (m, 2H), 3.59 (m, 1H), 3.37 (m, 2H), 2.66 (m, 1H), 2.22 (m, 1H), 1.75 (m, 1H), 2.10 (m, 1H), 1.83 (m, 1H), 1.74 (m, 1H), 1.65 (m, 1H), 1.42 (m, 1H);13C NMR (150 MHz, Methanol-d4): δ 153.3, 152.6, 151.7, 149.8, 144.0, 140.4, 133.7 (2C), 129.2, 122.4, 121.7, 113.7, 109.9, 104.9, 52.4, 49.0, 45.5, 40.7, 36.6, 30.8, 28.8, 26.0; HRESIMS m / z 433.1924, [M+H]+(calcd for C22H25N8S, 433.1923).

[0220] Synthesis of Compounds J and K from Acid Scaffold-D

[0221] Compounds J and K were prepared from Acid Scaffold-D, as shown in Scheme 12. A reaction vial containing a CHCl3 (2 mL) solution of Acid scaffold-D (1 equiv., 20 mg) was filled with N2and cooled down in an ice bath. Et3N (1 equiv., 12.4 µL) and a 40 mg / mL MeCN solution of N-bromosuccinimide (0.5 equiv., 0.2 mL) was added dropwise into the reaction vial. The resulting reaction mixture was stirred at 0 °C for 2 h, and subsequently was dried under vacuum. To the residue was added 4-(4,4,5,5-Tetramethyl-1,3,2- dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine (H-11, 54 mg), Xantphos Pd G4 (17 mg), Cs2CO3(116 mg), DMF (2 mL), and H2O (1 mL). The reaction vial was filled with N2and capped tightly. The reaction mixture was then stirred vigorously at 95 °C overnight before being dried under vacuum. The resulting residue was re-dissolved in DMSO followed by successive preparative HPLC purifications using a Gemini 5 μm NX-C18 column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10% to 100% MeCN in 0.1% TFA) and a Synergi 5 μm Hydro-RP column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10-100% MeCN in 0.1% TFA) to yield the products Compound J (0.4 mg, 1.5% yield) and Compound K (0.3 mg, 1.1% yield). Scheme 12: Synthesis of Compounds J and K

[0222] Preparation of 4-(1-methyl-1H-imidazol-5-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)- 3,4-dihydro-2H-1,4-thiazine (Compound J):4-(1-methyl-1H-imidazol-5-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H-1,4- thiazine (Compound J) was prepared from Acid scaffold-D as a light yellow solid.1H NMR (600 MHz, Methanol-d4): δ 8.87 (br s, 1H), 8.24 (br s, 1H), 7.63 (br s, 1H), 7.58 (s, 1H), 7.56 (br s, 1H), 7.48 (s, 1H), 7.15 (s, 1H), 4.03 (br s, 2H), 3.86 (s, 3H), 3.31 (m, 2H);13C NMR (150 MHz, Methanol-d4): δ 149.5, 140.9, 139.5, 135.6, 135.4, 134.2, 128.8, 121.9, 114.8, 113.5, 106.0, 104.2, 51.8, 33.3, 25.4; HRESIMS m / z 298.1130, [M+H]+(calcd for C15H16N5S, 298.1126).

[0223] Preparation of 4-(4-bromo-1-methyl-1H-imidazol-5-yl)-6-(1H-pyrrolo[2,3- b]pyridin-4-yl)-3,4-dihydro-2H-1,4-thiazine (Compound K):4-(4-bromo-1-methyl-1H-imidazol-5-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H- 1,4-thiazine (Compound K) was prepared from Acid scaffold-D as a yellow solid;1H NMR (600 MHz, Methanol-d4): δ 8.18 (br s, 1H), 7.76 (br s, 1H), 7.56 (s, 1H), 7.55 (s, 1H), 7.48 (s, 1H), 7.13 (s, 1H), 3.96 (m, 2H), 3.66 (s, 3H), 3.33 (m, 2H);13C NMR (150 MHz, Methanol- d4): δ 150.6, 140.0, 138.3, 137.9, 135.2, 133.4, 128.6, 121.2, 112.9, 110.8, 104.5, 103.0, 51.6, 32.3, 25.7; HRESIMS m / z 376.0234, [M+H]+(calcd for C15H15BrN5S, 376.0232).

[0224] General Procedure for Suzuki Coupling with Bromo Scaffold E:

[0225] Suzuki products were prepared from Bromo Scaffold-E, as shown in Scheme 13. To Bromo scaffold-E (1 equiv., x mg) was added 4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine (H-11, 2.5 equiv.), Xantphos Pd G4 (20 mol%), Cs2CO3(4 equiv.), DMF (x / 10 mL), and H2O (x / 20 mL). The reaction vial was filled with N2and capped tightly and the reaction mixture was then stirred vigorously at 95 °C overnight before it was dried under vacuum. The resulting residue was re-dissolved in DMSO followed by successive preparative HPLC purifications using a Gemini 5 μm NX-C18 column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10% to 100% MeCN in 0.1% TFA) and a Synergi 5 μm Polar-RP column (110 Å, 250 × 21.2 mm) with a flow rate of 10 mL / min (eluted with 10% to 100% MeCN in 0.1% TFA) to yield the Suzuki coupling product. Scheme 13: General Procedure for Suzuki Coupling with Bromo Scaffold-E

[0226] Preparation of 4-(pyrimidin-4-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro- 2H-1,4-thiazine (Compound L) from 4-(pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6- carboxylic acid.4-(pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 19% yield; white solid;1H NMR (600 MHz, DMSO-d6): δ 9.00 (br s, 1H), 8.70 (br s, 1H), 8.69 (s, 1H), 7.36 (s, 1H), 4.05 (m, 2H), 3.13 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 166.0, 158.5, 155.7, 153.6, 129.4, 106.9, 106.8, 43.5, 23.9; HRESIMS m / z 224.0492, [M+H]+(calcd for C9H10N3O2S, 224.0494).6-bromo-4-(pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine: 97% yield; pale white solid;1H NMR (600 MHz, DMSO-d6): δ 8.94 (br s, 1H), 8.60 (br s, 1H), 7.88 (s, 1H), 7.36 (s, 1H), 7.14 (br s, 1H), 4.19 (m, 2H), 3.34 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 157.7, 154.0, 149.7, 122.4, 105.2, 98.5, 42.0, 28.5; HRESIMS m / z 257.9703, [M+H]+(calcd for C8H9BrN3S, 257.9701).4-(pyrimidin-4-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H-1,4-thiazine (Compound L): 60% yield; light yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.13 (s, 1H), 8.98 (br s, 1H), 8.59 (br s, 1H), 8.34 (br s, 1H), 8.29 (s, 1H), 7.61 (t, J = 2.9 Hz, 1H), 7.42 (d, J = 6.3 Hz, 1H), 7.24 (d, J = 5.2 Hz, 1H), 6.82 (d, J = 2.9 Hz, 1H), 4.28 (m, 2H), 3.39 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 158.4, 153.9, 149.7, 147.1, 141.1, 139.2, 127.3, 122.6, 117.0, 115.6, 113.0, 105.7, 100.0, 43.6, 25.2; HRESIMS m / z 296.0969, [M+H]+(calcd for C15H14N5S, 296.0970).

[0227] Preparation of 4-(6-(benzyloxy)pyrimidin-4-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4- yl)-3,4-dihydro-2H-1,4-thiazine (Compound M) from 4-(6-(benzyloxy)pyrimidin-4-yl)-3,4- dihydro-2H-1,4-thiazine-6-carboxylic acid.4-(6-(benzyloxy)pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 47% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.56 (br s, 1H), 8.79 (s, 1H), 8.51 (s, 1H), 7.44 (d, J = 7.0 Hz, 2H), 7.38 (t, J = 7.0 Hz, 2H), 7.33 (t, J = 7.0 Hz, 1H), 6.54 (s, 1H), 5.41 (s, 2H), 3.92 (m, 2H), 3.06 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 170.1, 166.4, 159.9, 157.3, 136.9, 131.1, 128.5 (2C), 128.2 (2C), 128.1, 102.3, 89.7, 67.8, 43.8, 23.7; HRESIMS m / z 330.0913, [M+H]+(calcd for C16H16N3O3S, 330.0912).4-(6-(benzyloxy)pyrimidin-4-yl)-6-bromo-3,4-dihydro-2H-1,4-thiazine: 60% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.39 (s, 1H), 7.86 (br s, 1H), 7.43 (d, J = 7.0 Hz, 2H), 7.38 (t, J = 7.0 Hz, 2H), 7.33 (t, J = 7.0 Hz, 1H), 6.43 (s, 1H), 5.38 (s, 2H), 4.02 (m, 2H), 3.24 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 169.8, 159.4, 157.2, 136.4, 128.5 (2C), 128.1 (2C), 128.0, 123.7, 91.6, 87.6, 67.5, 42.0, 28.3; HRESIMS m / z 364.0117, [M+H]+(calcd for C15H15BrN3OS, 364.0119).4-(6-(benzyloxy)pyrimidin-4-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H-1,4- thiazine (Compound M): 51% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.37 (s, 1H), 8.65 (s, 1H), 8.51 (s, 1H), 8.31 (br s, 1H), 7.65 (d, J = 3.5 Hz, 1H), 7.45 (d, J = 7.1 Hz, 2H), 7.39 (t, J = 7.1 Hz, 2H), 7.33 (t, J = 7.1 Hz, 1H), 7.30 (d, J = 5.2 Hz, 1H), 6.89 (d, J = 3.5 Hz, 1H), 6.56 (s, 1H), 5.42 (s, 2H), 4.10 (m, 2H), 3.29 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 170.1, 160.2, 157.4, 144.4, 142.7, 138.6, 136.7, 128.7 (2C), 128.3 (2C), 128.2, 127.7, 126.4, 117.7, 112.5, 108.1, 101.0, 89.2, 67.8, 44.2, 25.0; HRESIMS m / z 402.1391, [M+H]+(calcd for C22H20N5OS, 402.1389).

[0228] Preparation of 4-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4-thiazin- 4-yl)-N,N,6-trimethylpyrimidin-2-amine (Compound N) from 4-(2-(dimethylamino)-6- methylpyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid.4-(2-(dimethylamino)-6-methylpyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 33% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.50 (s, 1H), 6.63 (s, 1H), 4.06 (m, 2H), 3.18 (s, 6H), 3.10 (m, 2H), 2.37 (s, 3H);13C NMR (150 MHz, DMSO-d6): δ 165.9, 160.5, 159.2, 155.1, 129.5, 107.6, 94.3, 43.3, 37.5, 24.0 (2C), 20.7; HRESIMS m / z 281.1072 [M+H]+(calcd for C12H17N4O2S, 281.1072).4-(6-bromo-2,3-dihydro-4H-1,4-thiazin-4-yl)-N,N,6-trimethylpyrimidin-2-amine: 20% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 7.74 (br s, 1H), 6.66 (s, 1H), 4.17 (m, 2H), 3.30 (m, 2H), 3.18 (s, 6H), 2.34 (s, 3H);13C NMR (150 MHz, DMSO-d6): δ 158.3, 158.0, 153.8, 122.7, 98.1, 93.9, 41.7, 37.7, 28.6 (2C), 19.8; HRESIMS m / z 315.0278, [M+H]+(calcd for C11H16BrN4S, 315.0279).4-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4-thiazin-4-yl)-N,N,6- trimethylpyrimidin-2-amine (Compound N): 15% yield; yellow solid;1H NMR (600 MHz,DMSO-d6): δ 12.15 (s, 1H), 8.30 (br s, 2H), 7.60 (s, 1H), 7.24 (d, J = 4.8 Hz, 1H), 6.80 (d, J = 3.4 Hz, 1H), 6.73 (s, 1H), 4.25 (m, 2H), 3.37 (m, 2H), 3.20 (s, 6H), 2.39 (s, 3H);13C NMR (150 MHz, DMSO-d6): δ 158.9, 157.6, 153.6, 147.6, 141.5, 141.5, 139.5, 127.6, 123.2, 117.4, 115.9, 113.4, 100.3, 95.1, 44.0, 38.2, 25.8 (2C), 20.0; HRESIMS m / z 353.1543, [M+H]+(calcd for C18H21N6S, 353.1548).

[0229] Preparation of 4-(4-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4- thiazin-4-yl)pyrimidin-2-yl)morpholine (Compound O) from 4-(2-morpholinopyrimidin-4-yl)- 3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid.4-(2-morpholinopyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 43% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.50 (s, 1H), 8.17 (d, J = 6.3 Hz, 1H), 6.56 (d, J = 6.3 Hz, 1H), 4.02 (m, 2H), 3.68 (m, 8H), 3.08 (m, 2H);13C NMR (150 MHz, DMSO- d6): δ 166.4, 159.5, 157.8, 154.3, 130.3, 106.0, 96.0, 66.1 (2C), 44.6 (2C), 43.7, 24.3; HRESIMS m / z 309.1023, [M+H]+(calcd for C13H17N4O3S, 309.1021).4-(4-(6-bromo-2,3-dihydro-4H-1,4-thiazin-4-yl)pyrimidin-2-yl)morpholine: 44% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.07 (d, J = 7.0 Hz, 1H), 7.77 (s, 1H), 6.69 (s, 1H), 4.17 (m, 2H), 3.69 (m, 8H), 3.30 (s, 2H);13C NMR (150 MHz, DMSO-d6): δ 158.5, 154.5, 148.5, 123.1, 98.3, 95.8, 65.9 (2C), 44.8 (2C), 42.3, 29.0; HRESIMS m / z 343.0226, [M+H]+(calcd for C12H16BrN4OS, 343.0228).4-(4-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4-thiazin-4-yl)pyrimidin-2- yl)morpholine (Compound O): 28% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.20 (s, 1H), 8.37 (br s, 1H), 8.29 (s, 1H), 8.14 (d, J = 6.8 Hz, 1H), 7.65 (t, J = 3.4 Hz, 1H), 7.25 (d, J = 5.2 Hz, 1H), 6.80 (d, J = 3.4 Hz, 1H), 6.71 (d, J = 6.8 Hz, 1H), 4.23 (m, 2H), 3.37 (m, 8H), 3.35 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 158.8, 154.9, 150.2, 146.4, 140.5, 140.1, 127.4, 123.7, 117.1, 114.0, 112.9, 100.1, 95.9, 65.6 (2C), 44.5 (2C), 43.8, 25.3; HRESIMS m / z 381.1499, [M+H]+(calcd for C19H21N6OS, 381.1498).

[0230] Preparation of 4-(4-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4- thiazin-4-yl)pyrimidin-2-yl)morpholine (Compound P) from 4-(6-methyl-2- morpholinopyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid.4-(6-methyl-2-morpholinopyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 55% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.50 (s, 1H), 8.57 (s, 1H), 6.29 (s, 1H), 3.95 (m, 2H), 3.64 (m, 8H), 3.03 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 168.2, 166.3, 160.5, 159.3, 131.1, 101.8, 93.8, 66.1 (2C), 43.9 (2C), 43.2, 24.3, 23.9; HRESIMS m / z 323.1173, [M+H]+(calcd for C14H19N4O3S, 323.1178).4-(4-(6-bromo-2,3-dihydro-4H-1,4-thiazin-4-yl)-6-methylpyrimidin-2-yl)morpholine: 40% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 7.72 (s, 1H), 6.62 (s, 1H), 4.14 (m, 2H), 3.69 (m, 8H), 3.28 (s, 2H), 2.31 (s, 3H);13C NMR (150 MHz, DMSO-d6): δ 159.5, 158.3, 154.5, 122.9, 97.2, 94.3, 65.6 (2C), 44.7 (2C), 41.9, 28.6, 20.6; HRESIMS m / z 357.0383, [M+H]+(calcd for C13H18BrN4OS, 357.0385).4-(4-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4-thiazin-4-yl)pyrimidin-2- yl)morpholine (Compound P): 28% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.22 (s, 1H), 8.33 (br s, 1H), 8.29 (s, 1H), 7.65 (t, J = 3.0 Hz, 1H), 7.25 (d, J = 5.2 Hz, 1H), 6.80 (d, J = 3.0 Hz, 1H), 6.68 (s, 1H), 4.22 (m, 2H), 3.72 (m, 8H), 3.34 (m, 2H), 2.35 (s, 3H);13C NMR (150 MHz, DMSO-d6): δ 160.5, 159.3, 155.1, 146.5, 140.6 (2C), 127.8, 124.2, 117.6, 114.0, 113.2, 100.5, 95.4, 66.0 (2C), 45.2 (2C), 44.0, 25.7, 21.1; HRESIMS m / z 395.1657, [M+H]+(calcd for C20H23N6OS, 395.1654).

[0231] Preparation of 4-(6-(tert-butyl)pyrimidin-4-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)- 3,4-dihydro-2H-1,4-thiazine (Compound Q) from 4-(6-(tert-butyl)pyrimidin-4-yl)-3,4- dihydro-2H-1,4-thiazine-6-carboxylic acid.4-(6-(tert-butyl)pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 95% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.79 (s, 1H), 8.77 (s, 1H), 7.06 (s, 1H), 4.07 (m, 2H), 3.11 (m, 2H), 1.30 (s, 9H);13C NMR (150 MHz, DMSO-d6): δ 175.8, 166.2, 159.1, 155.9, 130.3, 104.9, 101.0, 43.6, 37.4, 28.9 (3C), 23.9; HRESIMS m / z 280.1125, [M+H]+(calcd for C13H18N3O2S, 280.1120).6-bromo-4-(6-(tert-butyl)pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine: 52% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.74 (s, 1H), 7.95 (s, 1H), 7.01 (s, 1H), 4.20 (m, 2H), 3.31 (m, 2H), 1.31 (s, 9H);13C NMR (150 MHz, DMSO-d6): δ 172.3, 158.4, 154.6, 123.0, 99.8, 96.7, 42.0, 37.0, 28.7 (3C), 28.6; HRESIMS m / z 314.0324, [M+H]+(calcd for C12H17BrN3S, 314.0327).4-(6-(tert-butyl)pyrimidin-4-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H-1,4- thiazine (Compound Q): 29% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.29 (s, 1H), 8.85 (s, 1H), 8.53 (s, 1H), 8.32 (br s, 1H), 7.65 (t, J = 2.7 Hz, 1H), 7.29 (d, J = 5.1 Hz, 1H), 7.11 (s, 1H), 6.86 (s, 1H), 4.30 (m, 2H), 3.37 (m, 2H), 1.33 (s, 9H);13C NMR (150 MHz, DMSO-d6): δ 172.5, 159.0, 154.6, 145.4, 140.9, 139.6, 127.6, 124.3, 117.5, 113.0,112.8, 100.6, 100.4, 43.8, 36.9, 28.6 (3C), 25.1; HRESIMS m / z 352.1600, [M+H]+(calcd for C19H22N5S, 352.1596).

[0232] Preparation of 4-(6-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4- thiazin-4-yl)pyrimidin-4-yl)morpholine (Compound R) from 4-(6-morpholinopyrimidin-4-yl)- 3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid.4-(6-morpholinopyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 95% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.78 (s, 1H), 8.28 (s, 1H), 6.27 (s, 1H), 3.94 (m, 2H), 3.61(m, 8H), 3.05 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 166.5,162.6, 158.7, 156.5, 131.7, 100.9, 85.1, 65.9 (2C), 44.3 (2C), 43.7 (2C), 23.7; HRESIMS m / z 309.1020, [M+H]+(calcd for C13H17N4O3S, 309.1021).4-(6-(6-bromo-2,3-dihydro-4H-1,4-thiazin-4-yl)pyrimidin-4-yl)morpholine: 54% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.22 (s, 1H), 7.84 (s, 1H), 6.17 (s, 1H), 4.06 (m, 2H), 3.61 (m, 8H), 3.24 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 161.8, 157.9, 155.7, 124.1, 91.3, 83.0, 65.8 (2C), 44.4 (2C), 41.9, 28.4; HRESIMS m / z 343.0226, [M+H]+(calcd for C12H16BrN4OS, 343.0228).4-(6-(tert-butyl)pyrimidin-4-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H-1,4- thiazine (Compound R): 32% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.48 (s,1H), 8.76 (s, 1H), 8.32 (s, 1H), 8.30 (br s, 1H), 7.68 (t, J = 3.0 Hz, 1H), 7.30 (d, J = 5.6 Hz, 1H), 6.92 (d, J = 3.0 Hz, 1H), 6.33 (s, 1H), 4.14 (m, 2H), 3.63 (m, 8H), 3.30 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 162.4, 158.7, 156.4, 144.0, 142.8, 137.3, 127.8, 127.7, 117.9, 112.1, 106.5, 101.3, 84.8, 65.9 (2C), 44.3 (2C), 44.1, 24.8; HRESIMS m / z 381.1506, [M+H]+(calcd for C19H21N6OS, 381.1498).

[0233] Preparation of 4-(2-(tert-butyl)-6-methylpyrimidin-4-yl)-6-(1H-pyrrolo[2,3- b]pyridin-4-yl)-3,4-dihydro-2H-1,4-thiazine (Compound S) from 4-(2-(tert-butyl)-6- methylpyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid.4-(2-(tert-butyl)-6-methylpyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 90% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.74 (s, 1H), 7.11 (s, 1H), 4.06 (m, 2H), 3.13 (m, 2H), 2.45 (s, 3H), 1.35 (s, 9H);13C NMR (150 MHz, DMSO-d6): δ 172.6, 166.1, 163.9, 161.3, 158.6, 129.8, 102.2, 43.5, 38.8, 28.8 (3C), 23.9, 22.2; HRESIMS m / z 294.1275, [M+H]+(calcd for C14H20N3O2S, 294.1276).6-bromo-4-(2-(tert-butyl)-6-methylpyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine: 55% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 7.90 (br s, 1H), 7.17 (br s, 1H), 4.22 (m, 2H), 3.35 (m, 2H), 2.43 (s, 3H), 1.36 (s, 9H);13C NMR (150 MHz, DMSO-d6): δ 171.1, 160.7, 157.5, 122.5, 101.2, 98.2, 42.0, 38.5, 28.4, 28.3 (3C), 20.7; HRESIMS m / z 328.0481, [M+H]+(calcd for C13H19BrN3S, 328.0483).4-(2-(tert-butyl)-6-methylpyrimidin-4-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H- 1,4-thiazine (Compound S): 41% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.18 (s, 1H), 8.54 (br s, 1H), 8.29 (d, J = 5.3 Hz, 1H), 7.63 (t, J = 3.0 Hz, 1H), 7.27 (br s, 1H), 7.26 (d, J = 5.3 Hz, 1H), 6.84 (s, 1H), 4.29 (m, 2H), 3.41 (m, 2H), 2.52 (s, 3H), 1.39 (m, 9H);13C NMR (150 MHz, DMSO-d6): δ 170.6, 160.2, 158.1, 146.8, 140.8, 139.4, 127.3, 122.9, 116.9, 114.9, 112.9, 102.1, 100.0, 43.9, 38.5, 28.2 (3C), 25.1, 20.5; HRESIMS m / z 366.1758, [M+H]+(calcd for C20H24N5S, 366.1752).

[0234] Preparation of 4-(6-methylpyrimidin-4-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4- dihydro-2H-1,4-thiazine (Compound T) from 4-(6-methylpyrimidin-4-yl)-3,4-dihydro-2H-1,4- thiazine-6-carboxylic acid.4-(6-methylpyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 92% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.99 (br s, 1H), 8.67 (s, 1H), 7.37 (s, 1H), 4.04 (m, 2H), 3.13 (m, 2H), 2.45 (s, 3H), 2.54 (s, 3H);13C NMR (150 MHz, DMSO-d6): δ 165.9, 163.6, 158.7, 154.4, 129.5, 107.0, 105.7, 43.5, 23.8, 22.1; HRESIMS m / z 238.0658, [M+H]+(calcd for C10H12N3O2S, 238.0650).6-bromo-4-(6-methylpyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine: 22% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.81 (s, 1H), 7.84 (s, 1H), 7.28 (s, 1H), 4.19 (m, 2H), 3.34 (m, 2H), 2.43 (s, 3H);13C NMR (150 MHz, DMSO-d6): δ 160.1, 158.0, 153.3, 122.5, 103.6, 98.7, 41.9, 28.4, 20.7; ESIMS m / z 291.271.9855, [M+H]+(calcd for C9H11BrN3S, 271.9857).4-(6-methylpyrimidin-4-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H-1,4-thiazine (Compound T): 38% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.12 (s, 1H), 8.94 (s, 1H), 8.29 (s, 1H), 8.25 (br s, 1H), 7.61 (t, J = 3.0 Hz, 1H), 7.42 (s, 1H), 7.24 (d, J = 5.0 Hz, 1H), 6.80 (d, J = 3.0 Hz, 1H), 4.30 (m, 2H), 3.40 (m, 2H), 2.48 (s, 3H);13C NMR (150 MHz, DMSO-d6): δ 158.8, 152.3, 147.4, 141.3, 138.8, 127.3, 122.2, 117.1, 115.3, 113.1, 104.4, 100.0, 43.5, 25.3, 20.0; HRESIMS m / z 310.1129, [M+H]+(calcd for C16H16N5S, 310.1126).

[0235] Preparation of 4-(6-methylpyrimidin-4-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4- dihydro-2H-1,4-thiazine (Compound U) from 4-(6-(thiophen-3-yl)pyrimidin-4-yl)-3,4- dihydro-2H-1,4-thiazine-6-carboxylic acid.4-(6-(thiophen-3-yl)pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 68% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.61 (br s, 1H), 8.86 (s, 1H), 8.77 (s, 1H), 8.49 (d, J = 3.0 Hz, 1H), 7.89 (d, J = 5.4 Hz, 1H), 7.70 (dd, J = 3.0, 5.2 Hz, 1H), 7.56 (s, 1H), 4.10 (m, 2H), 3.14 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 166.3, 159.8, 159.1, 157.9, 140.1, 130.7, 127.8, 127.6, 126.5, 103.3, 100.9, 43.6, 23.8; HRESIMS m / z 306.0374, [M+H]+(calcd for C13H12N3O2S2, 306.0371).6-bromo-4-(6-(thiophen-3-yl)pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine: 41% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.68 (s, 1H), 8.49 (d, J = 3.0 Hz, 1H), 7.97 (br s, 1H), 7.89 (d, J = 5.1 Hz, 1H), 7.70 (dd, J = 3.0, 5.1 Hz, 1H), 7.48 (s, 1H), 4.21 (m, 2H), 3.31 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 158.5, 158.1, 157.3, 139.6, 127.6, 127.5, 126.5, 123.5, 99.4, 93.5, 41.7, 28.5; ESIMS m / z 339.9576, [M+H]+(calcd for C12H11BrN3S2, 339.9578).4-(6-methylpyrimidin-4-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H-1,4-thiazine (Compound U): 34% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.23 (s, 1H), 8.80 (s, 1H), 8.62 (br s, 1H), 8.50 (d, J = 2.8 Hz, 1H), 8.31(br s, 1H), 7.91 (d, J = 4.9 Hz, 1H), 7.72 (dd, J = 2.8, 4.9 Hz, 1H), 7.64 (br s, 1H), 7.60 (s, 1H), 7.29 (d, J = 4.9 Hz, 1H), 6.89 (s, 1H), 4.30 (m, 2H), 3.38 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 159.2, 158.0, 157.1, 145.5, 141.3, 139.6, 139.2, 128.0, 127.7, 127.4, 126.5, 124.9, 117.4, 112.7, 110.5, 100.6, 100.5, 43.7, 25.1; HRESIMS m / z 378.0851, [M+H]+(calcd for C19H16N5S2, 378.0847).

[0236] Preparation of 6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(6-(2,2,2- trifluoroethoxy)pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine (Compound V) from 4-(6-(2,2,2- trifluoroethoxy)pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid.4-(6-(2,2,2-trifluoroethoxy)pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 28% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.79 (s, 1H), 8.55 (s, 1H), 6.87 (s, 1H), 5.07 (q, J = 3.0 Hz, 1H), 3.95 (m, 2H), 3.08 (m, 2H);13C NMR (150 MHz, DMSO- d6): δ 168.6, 166.3, 160.3, 157.1, 130.7, 121.0-126.6 (q, 1C), 103.2, 89.8, 61.4-62.1 (q, 1C), 43.7, 23.7; HRESIMS m / z 322.0477, [M+H]+(calcd for C11H11F3N3O3S, 322.0473).6-bromo-4-(6-(2,2,2-trifluoroethoxy)pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine: 22% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.43 (s, 1H), 7.87 (s, 1H), 6.58 (s, 1H), 5.03 (q, J = 3.0 Hz, 1H), 4.05 (m, 2H), 3.26 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 168.7, 160.1, 157.4, 123.9, 121.5-127.0 (q, 1C), 93.0, 88.0, 61.7-62.3 (q, 1C), 42.4, 28.7; ESIMS m / z 355.9578, [M+H]+(calcd for C10H10BrF3N3OS, 355.9680).6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(6-(2,2,2-trifluoroethoxy)pyrimidin-4-yl)-3,4-dihydro- 2H-1,4-thiazine (Compound V): 45% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.24 (s, 1H), 8.58 (s, 1H), 8.54 (s, 1H), 8.30 (br s, 1H), 7.63 (s, 1H), 7.60 (s, 1H), 7.28 (d, J = 5.2 Hz, 1H), 6.87 (d, J = 3.5 Hz, 1H), 6.69 (s, 1H), 5.07 (q, J = 3.0 Hz, 1H), 4.13 (m, 2H), 3.31 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 168.5, 160.4, 157.1, 145.3, 141.5, 139.4, 127.4, 125.4, 121.1-126.6 (q, 1C), 117.3, 112.6, 109.1, 100.6, 88.9, 61.3-62.0 (q, 1C), 44.0, 25.0; HRESIMS m / z 394.0957, [M+H]+(calcd for C17H15F3N5OS, 394.0949).

[0237] Preparation of 4-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4-thiazin- 4-yl)-6-propylthieno[2,3-d]pyrimidine (Compound W) from 4-(6-propylthieno[2,3- d]pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid.4-(6-propylthieno[2,3-d]pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 26% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.62 (br s, 1H), 8.65 (s, 1H), 8.51 (s, 1H), 7.36 (s, 1H), 4.26 (m, 2H), 3.11 (m, 2H), 2.91 (t, J = 7.5 Hz, 2H), 1.69 (m, 2H), 0.95 (t, J = 7.3 Hz, 3H);13C NMR (150 MHz, DMSO-d6): δ 169.2, 166.1, 153.9, 151.6, 144.1, 133.0, 118.7, 117.2, 103.4, 46.0, 32.2, 24.1, 23.8, 13.4; HRESIMS m / z 322.0688, [M+H]+(calcd for C14H16N3O2S2, 322.0684).4-(6-bromo-2,3-dihydro-4H-1,4-thiazin-4-yl)-6-propylthieno[2,3-d]pyrimidine: 18% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.51 (s, 1H), 7.74 (s, 1H), 7.32 (s, 1H), 6.60 (s, 1H), 4.31 (m, 2H), 3.31 (m, 2H), 2.90 (t, J = 7.5 Hz, 2H), 1.70 (m, 2H), 0.94 (t, J = 7.3 Hz, 3H);13C NMR (150 MHz, DMSO-d6): δ 168.7, 153.3, 151.7, 142.9, 125.4, 117.6 (2C), 92.9, 44.4, 32.2, 28.9, 24.0, 13.5; ESIMS m / z 355.9878, [M+H]+(calcd for C13H15BrN3S2, 355.9891).4-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4-thiazin-4-yl)-6-propylthieno[2,3- d]pyrimidine (Compound W): 49% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.31 (s, 1H), 8.61 (s, 1H), 8.42 (s, 1H), 8.32 (br s, 1H), 7.61 (s, 1H), 7.40 (s, 1H), 7.31 (d, J = 4.9 Hz, 1H), 6.91 (d, J = 3.0 Hz, 1H), 4.45 (m, 2H), 3.34 (m, 2H), 2.88 (t, J = 7.5 Hz, 2H), 1.67 (m, 2H), 0.94 (t, J = 7.3 Hz, 3H);13C NMR (150 MHz, DMSO-d6): δ 169.0, 154.2, 151.6, 144.7, 143.5, 142.0, 138.9, 127.7, 127.4, 118.5, 117.7, 117.6, 112.4, 109.1, 100.9,46.4, 32.2, 25.3, 24.0, 13.5; HRESIMS m / z 394.1170, [M+H]+(calcd for C20H20N5S2, 394.1160).

[0238] Preparation of 6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(6- (trifluoromethyl)pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine (Compound X) from 4-(6- (trifluoromethyl)pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid.4-(6-(trifluoromethyl)pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 60% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.83 (br s, 1H), 8.95 (s, 1H), 8.76 (s, 1H), 7.62 (s, 1H), 4.09 (m, 2H), 3.13 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 166.0, 159.5, 158.4, 154.4-153.7 (q, 1C), 129.5, 123.6-118.1 (q, 1C), 106.5, 102.9-102.8 (q, 1C), 43.6, 23.9; HRESIMS m / z 292.0365, [M+H]+(calcd for C10H9F3N3O2S, 292.0368).6-bromo-4-(6-(trifluoromethyl)pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine: 21% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.81 (s, 1H), 7.95 (br s, 1H), 7.59 (br s, 1H), 6.58 (br s, 1H), 4.18 (m, 2H), 3.30 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 158.5, 158.4, 153.8-153.2 (q, 1C), 123.8-118.3 (q, 1C), 122.8, 101.6, 95.9, 44.0, 28.5; ESIMS m / z 325.9570, [M+H]+(calcd for C9H8BrF3N3S, 325.9575).6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(6-(trifluoromethyl)pyrimidin-4-yl)-3,4-dihydro-2H-1,4- thiazine (Compound X): 17% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.06 (s, 1H), 8.90 (s, 1H), 8.44 (br s, 1H), 8.29 (br s, 1H), 7.62 (s, 1H), 7.60 (s, 1H), 7.23 (d, J = 4.7 Hz, 1H), 6.83 (d, J = 3.5 Hz, 1H), 4.27 (m, 2H), 3.35 (m, 2H);13C NMR (150 MHz, DMSO- d6): δ 159.3, 158.5, 153.9-153.2 (q, 1C), 147.2, 141.1, 139.5, 127.1, 123.8-118.3 (q, 1C), 123.2, 117.1, 113.0 (2C), 102.3-102.2 (q, 1C), 100.1, 43.5, 25.2; HRESIMS m / z 364.0847, [M+H]+(calcd for C16H13F3N5S, 364.0844).

[0239] Preparation of 4-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4-thiazin- 4-yl)thieno[2,3-d]pyrimidine (Compound Y) from 4-(thieno[2,3-d]pyrimidin-4-yl)-3,4- dihydro-2H-1,4-thiazine-6-carboxylic acid.4-(thieno[2,3-d]pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 27% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.66 (br s, 1H), 8.71 (s, 1H), 8.54 (s, 1H), 7.89 (d, J = 6.1 Hz, 1H), 7.63 (d, J = 6.1 Hz, 1H), 4.30 (m, 2H), 3.13 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 169.8, 166.1, 155.0, 152.2, 132.9, 125.8, 120.7, 118.2, 104.0, 45.9, 24.2; HRESIMS m / z 280.0217, [M+H]+(calcd for C11H10N3O2S2, 280.0214).4-(6-bromo-2,3-dihydro-4H-1,4-thiazin-4-yl)thieno[2,3-d]pyrimidine: 31% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.58 (s, 1H), 7.79 (d, J = 6.1 Hz, 1H), 7.77 (s, 1H), 7.59 (d, J = 6.1 Hz, 1H), 4.34 (m, 2H), 3.32 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 169.3, 154.2, 152.2, 125.4, 124.5, 121.1, 117.1, 93.7, 44.3, 29.0; ESIMS m / z 315.9407, [M+H]+(calcd for C10H9BrN3S2, 315.9421).4-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4-thiazin-4-yl)thieno[2,3- d]pyrimidine (Compound Y): 53% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.21(s, 1H), 8.67 (s, 1H), 8.40 (s, 1H), 8.29 (br s, 1H), 7.81 (d, J = 6.2 Hz, 1H), 7.65 (d, J = 6.2 Hz, 1H), 7.59 (s, 1H), 7.28 (d, J = 5.3 Hz, 1H), 6.89 (d, J = 2.8 Hz, 1H), 4.48 (m, 2H), 3.36 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 169.6, 155.1, 152.2, 145.5, 141.2, 139.7, 127.3, 127.0, 125.0, 121.1, 117.9, 117.3, 112.5, 110.0, 100.6, 46.1, 25.4; HRESIMS m / z 352.0697, [M+H]+(calcd for C17H14N5S2, 352.0691).

[0240] Preparation of 4-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4-thiazin- 4-yl)furo[2,3-d]pyrimidine (Compound Z) from 4-(furo[2,3-d]pyrimidin-4-yl)-3,4-dihydro- 2H-1,4-thiazine-6-carboxylic acid.4-(furo[2,3-d]pyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 39% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.69 (br s, 1H), 8.75 (s, 1H), 8.58 (s, 1H), 8.11 (d, J = 2.6 Hz, 1H), 7.21 (d, J = 2.6 Hz, 1H), 4.28 (m, 2H), 3.14 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 167.7, 166.2, 154.3, 152.5, 143.8, 131.5, 105.9, 104.1, 103.4, 45.3, 24.0; HRESIMS m / z 264.0440, [M+H]+(calcd for C11H10N3O3S, 264.0443).4-(6-bromo-2,3-dihydro-4H-1,4-thiazin-4-yl)furo[2,3-d]pyrimidine: 19% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.45 (s, 1H), 8.03 (d, J = 2.6 Hz, 1H), 7.95 (s, 1H), 7.20 (d, J = 2.6 Hz, 1H), 4.31 (m, 2H), 3.33 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 167.5,153.6, 152.5, 142.9, 124.0, 106.0, 101.9, 93.5, 43.9, 28.6; ESIMS m / z 297.9653, [M+H]+(calcd for C10H9BrN3OS, 297.9650).4-(6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-2,3-dihydro-4H-1,4-thiazin-4-yl)furo[2,3-d]pyrimidine (Compound Z): 70% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.23 (s, 1H), 8.63 (s, 1H), 8.55 (s, 1H), 8.30 (br s, 1H), 8.07 (d, J = 2.5 Hz, 1H), 7.62 (s, 1H), 7.29 (d, J = 5.2 Hz, 1H), 7.25 (d, J = 2.6 Hz, 1H), 6.89 (d, J = 3.5 Hz, 1H), 4.46 (m, 2H), 3.37 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 167.6, 154.4, 152.6, 145.6, 143.2, 141.2, 139.7, 127.4, 125.8, 117.2, 112.5, 110.0, 106.1, 102.9, 100.5, 457.7, 25.2; HRESIMS m / z 336.0925, [M+H]+(calcd for C17H14N5OS, 336.0919).

[0241] Preparation of 4-(5-fluoropyrimidin-4-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4- dihydro-2H-1,4-thiazine (Compound AA) from 4-(5-fluoropyrimidin-4-yl)-3,4-dihydro-2H- 1,4-thiazine-6-carboxylic acid.4-(5-fluoropyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine-6-carboxylic acid: 13% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 12.69 (br s, 1H), 8.67 (s, 1H), 8.64 (s, 1H), 8.38 (s, 1H), 4.12 (m, 2H), 3.10 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 166.0, 153.5-153.4 (d), 148.4-146.7 (d), 148.0 (d), 132.5-132.4 (d), 104.0, 44.9-44.8 (d), 23.9; HRESIMS m / z 242.0394, [M+H]+(calcd for C9H9FN3O2S, 242.0400).6-bromo-4-(5-fluoropyrimidin-4-yl)-3,4-dihydro-2H-1,4-thiazine: 21% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 8.59 (br s, 2H), 7.59 (s, 1H), 4.18 (m, 2H), 3.30 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 158.3-158.1 (d), 153.3 (d), 147.4 (d), 144.1-144.0 (d), 124.7-124.6 (d), 93.3, 43.3-43.2 (d), 28.6; ESIMS m / z 275.9610, [M+H]+(calcd for C8H8BrFN3S, 275.9606).4-(5-fluoropyrimidin-4-yl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,4-dihydro-2H-1,4-thiazine (Compound AA): 28% yield; yellow solid;1H NMR (600 MHz, DMSO-d6): δ 11.76 (s, 1H), 8.30 (br s, 2H), 7.98 (br s, 1H), 7.76 (s, 1H), 7.21 (d, J = 3.5 Hz, 1H), 6.86 (s, 1H), 6.49 (s, 1H), 3.92 (m, 2H), 2.95 (m, 2H);13C NMR (150 MHz, DMSO-d6): δ 156.9, 153.4 (d), 148.2 (d), 146.1, 144.5-144.3 (d), 140.4, 140.2, 127.2, 126.3, 117.2, 112.7, 110.1, 100.3, 44.7-44.6 (d), 25.1; HRESIMS m / z 314.0882, [M+H]+(calcd for C15H13FN5S, 314.0876). EXAMPLE 3

[0242] This example provides a comparative analysis of the computational docking score in the ATP binding site of chimeric J-PKAcα fusion protein exhibited by the reverse thiazine compounds of the present invention relative to normal thiazine compounds.

[0243] In particular, 18 analogues consisting of 9 normal thiazine analogues and 9 reverse thiazine analogues were analyzed to determine the computational docking score in the ATP binding site of chimeric J-PKAcα fusion protein. The difference ∆DS (i.e., N-M) between the normal thiazine analog (N) and the reverse thiazine analog (M) is plotted in Table 1. A positive ∆DS indicates that the reverse thiazine analog binds more favorably in the ATP binding site of chimeric J-PKAcα fusion protein and a higher ∆DS indicates how significant the strength in binding is.Table 1. Computational Docking Score for ATP Binding Site of Chimeric J-PKAcα Fusion Protein

[0244] As is apparent from the results set forth in Table 1, the reverse thiazine analogues bind more favorably in the ATP binding site of chimeric J-PKAcα fusion protein than the normal thiazine analogues in all instances. EXAMPLE 4

[0245] This example provides the IC50value for inhibitory activity against an enzymatically active chimeric protein, J-PKAcα, which is found in almost all FLHCC patients with greater than 10-fold overexpression of the fusion kinase in tumor cells relative to wild-type PKA (wt-PKA) expression in adjacent normal liver tissue.

[0246] Compounds were tested to determine an IC50value for inhibitory activity against RIα2:JPKAcα2chimeric kinase holoenzyme. A 3× dose-response curve was set up over a final compound concentration range of 0−10 μM containing 1 μM cAMP, 50 μM ATP, and 0.4% DMSO in 100 mM Tris−HCl pH 7.5 (all final concentrations). A no cAMP / ATP control was also included for background normalization. Using 12-channel multichannel pipettes, quadruplicate reactions were initiated by the addition of 20 μL of compound / cAMP / ATP solution to reaction wells containing 40 μL of 1.5× concentration PKA holoenzyme (chimeric) and biotinylated substrate protein (0.5 nM Chimeric Kinase Holoenzyme or 0.66 nM wt-kinase holoenzyme, 50 μM biotinylated substrate, in kinase buffer. After initiation by addition, the reaction was allowed to proceed for 45 min prior tothe addition of 15 μL of 0.5 M EDTA to quench the reactions. Quenched reactions were then transferred to prepared assay binding plates and the ELISA was developed as described above for the primary screening assay. For each reaction well, observed RFU were converted to normalized % activity measurements as described above using the No cAMP / ATP wells as the low control and the vehicle control (0 μM) as the high control. Representative % JPKAcα Activity curves for Compounds D-L are set forth in FIGs.1-9, respectively.

[0247] The % normalized activity measures were then fit to the following equation using a nonlinear regression least-squares fit with a variable slope (GraphPad Prism Software, San Diego, CA) an IC50value was calculated using the following formula.The mean (from three dose response curves) J-PKAcα IC50and WT-PKAcα IC50values are set forth in Table 2. Table 2. J-PKAcα IC50and WT-PKAcα IC50values* A < 0.1 µM; B is 0.1 µM – 1 µM; C is 1.01 µM – 10 µM; D is > 10 µM

[0248] In addition, to confirm the computational results provided in Example 3, the IC50value for inhibitory activity against an enzymatically active chimeric protein, J-PKAcα, was determined for 4 analogues consisting of 2 normal thiazine analogues and 2 reverse thiazine analogues. In particular, the J-PKAcα IC50values for the 4 analogues are set forth in Table 3. Table 3. J-PKAcα IC50for Normal Thiazine and Reverse Thiazine Analogues

[0249] As is apparent from the results set forth in Table 3, the reverse thiazine analogues Compound L and Compound J were more potent inhibitors of J-PKAcα than their corresponding normal thiazine counterparts, thereby confirming the computational results of Example 3. EXAMPLE 5

[0250] This example demonstrates the ability to reduce the viability of HCT-116 colon cancer cells exhibited by the reverse thiazine compounds of the present invention.

[0251] Compounds D-L were tested for their ability to inhibit the growth of HCT-116 colon cancer cells. In particular, wild-type (WT) and TOPO3β knockout (KO) HCT-116 cells were subject to a cytotoxicity assay at concentrations ranging from 0.5-30 µM.Representative HCT-116 cell viability curves for Compounds D, H, I, and L are set forth in FIGs.10-13, respectively.

[0252] The % normalized activity measures were then fit to the following equation using a nonlinear regression least-squares fit with a variable slope (GraphPad Prism Software, San Diego, CA) an IC50value was calculated using the following formula.The mean (from three dose response curves) TOPO3β (KO) IC50and TOPO3β (WT) IC50values are set forth in Table 4. Table 4. TOPO3β (KO) IC50and TOPO3β (WT) HCT-116 cells IC50valuesEXAMPLE 6

[0253] This example provides kinome profiling results of reverse thiazine compounds of the present invention.

[0254] Compounds B and J were kinome profiled. The study included 375 kinases and the 50 most sensitive kinases to Compounds B and J are listed in Table 5. Table 5. Kinome Profiling of Compounds B and J

[0255] As is apparent from the results set forth in Table 5, Compounds B and J significantly inhibit a series of kinases in the AGC (e.g., protein kinase A, protein kinase G, and protein kinase C) kinase family such as, for example, ROCK (Rho-associated protein kinase). EXAMPLE 7

[0256] This example demonstrates the ability to inhibit phosphorylation of protein kinase A (PKA) substrate cAMP response element-binding protein (CREB) exhibited by the reverse thiazine compounds of the present invention.

[0257] Inhibition of CREB phosphorylation at position Ser133 by Compounds J and L was measured using a Western Blot study with a Ser113-phospho-CREB specific antibody. The normalized homogeneous time resolved fluorescence (HTRF) percentage of the Western Blot study for Compounds J and L is plotted in FIGs.14 and 15, respectively.

[0258] As is apparent from the results set forth in FIGs.14 and 15, Compounds J and L showed concentration dependent inhibition of PKA in cells, as evidenced by their ability to reduce phosphorylation at position Ser133 of PKA substrate CREB. EXAMPLE 8

[0259] This example demonstrates the ability to inhibit CLK2 activity exhibited by the reverse thiazine compounds of the present invention.

[0260] Inhibition of CLK2 activity by Compounds A, C, D, E, F, H, I, J and L was assessed. Briefly, the compounds were tested in single dose duplicate mode at a concentration of 0.05 μM. Control compound, Staurosporine, was tested in 10-dose IC50mode with 4-fold serial dilution starting at 20 μM. Reactions were carried out at 10 μM ATP. The average CLK2 % enzyme activity is set forth in Table 6. Table 6. CLK2 Inhibition

[0261] As is apparent from the results set forth in Table 6, most of the compounds exhibited some level of CLK2 enzyme inhibition with Compounds J and L exhibiting significant inhibition of CLK2 enzyme activity. EXAMPLE 9

[0262] This example provides the NCI-60 human tumor cell line screen results for reverse thiazine compounds of the present invention.

[0263] Compounds B, C, J, L, T, Y, and Z were subjected to HTS384 NCI-60 human tumor cell line screen. The GI50(growth inhibition 50%), TGI (total growth inhibition), and LC50(lethal concentration 50%) results are shown below in Tables 7-9, respectively. Table 7. NCI-60 Human Tumor Cell Line GI50Table 8. NCI-60 Human Tumor Cell Line TGITable 9. NCI-60 Human Tumor Cell Line LC50

[0264] As is apparent from the results set forth in Tables 7-9, Compounds B, C, J, L, T, Y, and Z exhibited a broad range of activity (e.g., growth inhibition and cytotoxicity) against the NCI-60 human tumor cell lines, evidencing the ability of the reverse thiazine compounds of the present invention to be useful for the treatment or prevention cancer.

[0265] Compound C showed the most potent antiproliferative activity across the 60-cell- line panel (mean GI50= 1.4 µM). This compound showed potent cytotoxic activities against the leukemia cell lines MOLT-4 and SR, killing 50% of the initially plated cells (LC50) at concentrations of 780 and 600 nM, respectively. Compounds B, L, T, Y, and Z showed moderate antiproliferative activities with GI50values in the range of 3.7-17.7 µM.EXAMPLE 10

[0266] This example provides the results of a spheroid cancer cell viability study for reverse thiazine compounds of the present invention.

[0267] Using a Multi-cell Type Spheroid Assay, compounds B and L were screened against patient-derived pancreatic, non-small cell lung, and colon cancer cells. In this assay, multi-cell type tumor spheroids were grown from a mixture of malignant cells, endothelial cells, and mesenchymal stem cells. The cell viability upon compound treatment was assessed using the CellTiter-Glo 3D assay. Both compounds were tested in a dose-reposnse manner (2.5 nM – 25 μM) and the results are shown below in Table 10. Table 10: Spheroid Cancer Cell Viability IC50

[0268] As is apparent from the data in Table 10, Compounds B and L inhibited patient- derived pancreatic cancer cell viability in a spheroid cancer cell viability assay. The median IC50value of Compound B was 25 µM and the median IC50value of Compound L was 16 µM. EXAMPLE 11

[0269] This example provides the results of X-ray diffraction experiments that show the co-crystal structure of a compound of the present invention with J-PKAcα.

[0270] Crystallization and X-ray Data Collection. WT PKAcα and J-PKAcα proteins were concentrated to ~2-3 mg / mL and mixed with PKI (5-24) peptide (TOCRIS) and Compound L to a final concentration of 0.6 mM and 2.5 mM, respectively. The mixtures were incubated for 10 minutes at room temperature before crystallization. Optimized crystals were obtained under the conditions 200 mM lithium sulfate, 100 mM HEPES pH 7.6, and 25% PEG3350 by hanging drop diffusion method at 4 °C. For data collection, crystals were soaked in the solvent identical to the reservoir solution with 25% v / v glycerol as cryoprotectant and flash frozen into liquid nitrogen. All diffraction data were collected at 22ID beamline of the Advanced Photon Source (APS), Argonne National Laboratory (ANL) or in the Advanced light source (ALS), Lawerence Berkeley National Laboratory.

[0271] Structure Determination. All diffraction data sets were processed with XDS program, and Pointless program was used for the space group determination. Initial phases were obtained by molecular replacement in PHASER using the structure of WT PKAcα (PDB ID: 6BYS) and J-PKAcα (PDB ID: 4WB7) as searching models. The models were built with COOT and refined by PHENIX. The final structure of J-PKAcα:PKI : Compound L was determined to be at 2.77 Å resolution.

[0272] The pyrimidine N1 and N3 of Compound L formed a three-component H-bond complex with Lys127 and the DFG-1 residue Thr238. This new binding mode affords improved binding affinity and stability of the Compound L : J-PKAcα, contributing to the significantly increased biochemical activity of Compound L : J-PKAcα. FIG.16 shows a ribbon diagram of the structure of the co-crystallized complex of J-PKAcα and Compound L. FIG.17 is a diagram showing the four J-PKAcα residues that mediate the binding affinity of Compound L, i.e., Val 178, Glu 176, Thr 238, and Lys 127.EXAMPLE 12

[0273] This example shows that a compound of the present invention provides anti- malarial activity.

[0274] Compound I was included in a malaria asexual assay as part of a screening of a aplithianine library with 581 compounds. The Pf 3D7 parasite was treated with compounds at 37 °C for 72 h. The viability of the parasite was measured by addition of DAPI followed by dark incubation and high-resolution imaging. Compound I exhibited anti-malarial activity with an IC50of 4.3 µM. FIG.18 is a graph showing the dose-response of Compound I in the malaria asexual assay. EXAMPLE 13

[0275] This example shows that compounds of the present invention inhibit CLK- dependent RNA splicing.

[0276] The mRNA transcript for CLK kinases contains a “poison exon” that must be spliced out in order for the transcript to be fully processed into a protein through translation. The splicing out of this exon requires CLK activity to regulate proper and specific spliceosomal activity. In the RNA splicing assay, HEK293T cells were treated with compounds and the endogenous splicing of CLK1 exon 4 was monitored and measured by RT-PCR. As seen in FIG.19, Compounds B and J inhibited CLK-dependent RNA splicing in the HEK293T cell line by > 70% at 150 nM. Cirtuvivint and DMSO were used as controls. Cirtuvivint is a potent and orally active CDC-like kinase (CLK) inhibitor.

[0277] All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein.

[0278] The use of the terms “a” and “an” and “the” and “at least one” and similar referents in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The use of the term “at least one” followed by a list of one or more items (for example, “at least one of A and B”) is to be construed to mean one item selected from the listed items (A or B) or any combination of two or more of the listed items (A and B), unless otherwise indicated herein or clearly contradicted by context. The terms “comprising,” “having,” “including,” and “containing”are to be construed as open-ended terms (i.e., meaning “including, but not limited to,”) unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0279] Preferred embodiments and aspects of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations of those preferred embodiments may become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventors expect skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

Claims

1. CLAIM(S):

1. A compound of formula (I)whereinis a single or double bond, X1and X2are each independently CH, CR4, or N; X3is S, S=O, or S(=O)2; R1is H or –NR2R3; R2is H, C1-C3alkyl, or an aryl; R3is H, C1-C3alkyl, or an aryl; R4is C1-C3alkyl; and E is C3-C8heterocycloalkyl, C8-C10bicycloalkyl, C4-C10biheterocycloalkyl, aryl, or heteroaryl, each of which is optionally substituted with one or more substituents selected from C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, trifluoromethyl, C1-C6alkoxy, arylated C1-C6alkoxy, trifluoromethylated C1-C6alkoxy, -C(O)OH, -C(O)O-(C1-C6alkyl), -C(O)-(C1-C6alkyl), -(C1-C3alkyl)-C(O)OH, -(C1-C3alkyl)-C(O)O-(C1-C6alkyl), -(C1-C3alkyl)-C(O)-(C1- C6alkyl), -NH-C(O)OH, -NH-C(O)O-(C1-C6alkyl), -NH-C(O)-(C1-C6alkyl), -(C1-C3alkyl)-NH-C(O)OH, -(C1-C3alkyl)-NH-C(O)O-(C1-C6alkyl), -(C1-C3alkyl)-NH-C(O)-(C1-C6alkyl), -(C1-C6alkyl)-O-(C1-C6alkyl), -O-(C1-C6alkyl)-O-(C1-C6alkyl), -(C1-C6alkyl)-OH, - (C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-COOH, -(C1-C6alkyl)-C(O)O-(C1-C6alkyl), -(C1-C6alkyl)-(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-NH2, -(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)- (C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)-heterocycloalkyl, -(C1-C6alkyl)-heterocycloalkyl- (C1-C6alkyl), -(C1-C6alkyl)-C(O)-heterocycloalkyl, -(C1-C6alkyl)-C(O)-heterocycloalkyl- (C1-C6alkyl), halo, nitro, hydroxy, amino, C1-C6alkylamino, di-C1-C6alkyl-amino; -NH- aryl, -NH-heterocycloalkyl, -NH-heterocycloalkyl-(C1-C6alkyl), -C1-C6haloalkyl, C3-C8cycloalkyl, C3-C8heterocycloalkyl, fused C3-C8cycloalkyl, spiro C3-C8cycloalkyl, fused C3- C8heterocycloalkyl, spiro C3-C8heterocycloalkyl, aryl, heteroaryl, fused aryl, fused heteroaryl, -CN, -(C1-C3alkyl)-CN, carbonyl, amino acid, amino acid methyl ester, and a combination thereof, or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein E is a carbon bound C3-C8heterocycloalkyl, aryl, or heteroaryl, each of which is optionally substituted with one or more substituents selected from C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, trifluoromethyl, C1-C6alkoxy, arylated C1-C6alkoxy, trifluoromethylated C1-C6alkoxy, -(C1-C6alkyl)-OH, -(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-COOH, -(C1-C6alkyl)- NH2, -(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)-(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl) heterocycloalkyl, halo, nitro, hydroxy, amino, C1-C6alkylamino, di-C1-C6alkyl-amino; -NH-aryl, C1-C6haloalkyl, C3-C8cycloalkyl, C3-C8heterocycloalkyl, fused C3- C8cycloalkyl, fused C3-C8heterocycloalkyl, aryl, heteroaryl, fused aryl, fused heteroaryl, - CN, -(C1-C3alkyl)-CN, carbonyl, and a combination thereof, or a pharmaceutically acceptable salt thereof.

3. The compound of claim 1, wherein E is a carbon bound C3-C8heteroaryl, which is optionally substituted with one or more substituents selected from C1-C6alkyl, C2- C6alkenyl, C2-C6alkynyl, trifluoromethyl, C1-C6alkoxy, arylated C1-C6alkoxy, trifluoromethylated C1-C6alkoxy, -(C1-C6alkyl)-OH, -(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-COOH, -(C1-C6alkyl)-NH2, -(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)-(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl) heterocycloalkyl, halo, nitro, hydroxy, amino, C1-C6alkylamino, di-C1-C6alkyl-amino; -NH-aryl, C1-C6haloalkyl, C3-C8cycloalkyl, C3-C8heterocycloalkyl, fused C3-C8cycloalkyl, fused C3-C8heterocycloalkyl, aryl, heteroaryl, fused aryl, fused heteroaryl, -CN, -(C1-C3alkyl)-CN, carbonyl, and a combination thereof, or a pharmaceutically acceptable salt thereof.

4. The compound of any one of claims 1-3, whereinis a double bond, or a pharmaceutically acceptable salt thereof.

5. The compound of any one of claims 1-4, wherein R1is H, or a pharmaceutically acceptable salt thereof.

6. The compound of any one of claims 1-5, wherein the compound of formula (I) is of formula (Ia): ,or a pharmaceutically acceptable salt thereof.

7. The compound of any one of claims 1-6, wherein the compound of formula (I) is of formula (Ib), (Ic), (Id), (Ie), or (1f):or a pharmaceutically acceptable salt thereof.

8. The compound of any one of claims 1-7, wherein E is selected from:,each of which is optionally substituted with one or more substituents selected from C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, trifluoromethyl, C1-C6alkoxy, arylated C1-C6alkoxy, trifluoromethylated C1-C6alkoxy, -C(O)OH, -C(O)O-(C1-C6alkyl), -C(O)-(C1-C6alkyl), - (C1-C3alkyl)-C(O)OH, -(C1-C3alkyl)-C(O)O-(C1-C6alkyl), -(C1-C3alkyl)-C(O)-(C1-C6alkyl), -NH-C(O)OH, -NH-C(O)O-(C1-C6alkyl), -NH-C(O)-(C1-C6alkyl), -(C1-C3alkyl)- NH-C(O)OH, -(C1-C3alkyl)-NH-C(O)O-(C1-C6alkyl), -(C1-C3alkyl)-NH-C(O)-(C1-C6alkyl), -(C1-C6alkyl)-O-(C1-C6alkyl), -O-(C1-C6alkyl)-O-(C1-C6alkyl), -(C1-C6alkyl)-OH, - (C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-COOH, -(C1-C6alkyl)-C(O)O-(C1-C6alkyl), -(C1-C6alkyl)-(C1-C6cycloalkyl)-OH, -(C1-C6alkyl)-NH2, -(C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)- (C1-C6cycloalkyl)-NH2, -(C1-C6alkyl)-heterocycloalkyl, -(C1-C6alkyl)-heterocycloalkyl- (C1-C6alkyl), -(C1-C6alkyl)-C(O)-heterocycloalkyl, -(C1-C6alkyl)-C(O)-heterocycloalkyl- (C1-C6alkyl), halo, nitro, hydroxy, amino, C1-C6alkylamino, di-C1-C6alkyl-amino; -NH- aryl, -NH-heterocycloalkyl, -NH-heterocycloalkyl-(C1-C6alkyl), -C1-C6haloalkyl, C3-C8cycloalkyl, C3-C8heterocycloalkyl, fused C3-C8cycloalkyl, spiro C3-C8cycloalkyl, fused C3- C8heterocycloalkyl, spiro C3-C8heterocycloalkyl, aryl, heteroaryl, fused aryl, fusedheteroaryl, -CN, -(C1-C3alkyl)-CN, carbonyl, amino acid, amino acid methyl ester, and a combination thereof, or a pharmaceutically acceptable salt thereof.

9. The compound of claim 1, wherein the compound of formula (I) isor a pharmaceutically acceptable salt thereof.

10. A pharmaceutical composition comprising a compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, and a pharmaceutical carrier.

11. A method of inhibiting kinase activity in a subject, the method comprising administering to the subject a compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 10 to the subject.

12. The method of claim 11, wherein the kinase is PKA, PKA / DNAJ, cAMP- PKA, PKG1a, PKG1b, PKG2, PfPKG, PKC-θ, PKC-nu, PKC-d, PKC-eta, PKC-g, STK39, CLK1, CLK2, CLK3, CLK4, DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, LATS1, or LATS2.

13. The method of claim 11, wherein the kinase is PKA, PKA / DNAJ, or cAMP- PKA.

14. The method of claim 11, wherein the kinase is PKG1a, PKG1b, PKG2, or PfPKG.

15. The method of claim 11, wherein the kinase is DYRK1A, DYRK1B, DYRK2, DYRK3, or DYRK4.

16. The method of claim 11, wherein the kinase is CLK1, CLK2, CLK3, or CLK4.

17. A method of suppressing the immune system in a subject, the method comprising administering to the subject a compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 10 to the subject.

18. A method of preventing organ rejection in a subject, the method comprising administering to the subject a compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 10 to the subject.

19. A method of treating cancer in a subject, the method comprising administering to the subject a compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 10 to the subject.

20. The method of claim 19, wherein the cancer is fibrolamellar carcinoma (FLC).

21. The method of claim 19, wherein the cancer is fibrolamellar hepatocellular carcinoma (FL-HCC).

22. The method of claim 19, wherein the cancer is gastric cancer, colon cancer, pancreatic cancer, prostate cancer, breast cancer, lung cancer, or a glioma.

23. A method of modulating mRNA splicing in a subject, the method comprising administering to the subject a compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 10 to the subject.

24. A method of treating diabetic neuropathic pain in a subject, the method comprising administering to the subject a compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 10 to the subject.

25. A method of treating malaria in a subject, the method comprising administering to the subject a compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 10 to the subject.

26. A method of treating an infection associated with a protozoa in a subject, the method comprising administering to the subject a compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 10 to the subject.

27. A method of treating neurodegenerative disease in a subject, the method comprising administering to the subject a compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 10 to the subject.

28. A method of treating Down Syndrome in a subject, the method comprising administering to the subject a compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 10 to the subject.

29. The method of any one of claims 11-28, wherein the subject is human.

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