TEC kinase agonists and their use in enhancing lymphocyte activation

Abstract

The present disclosure provides for compounds and methods of treating cancer, methods of increasing lymphocyte activation, and methods of activating TEC kinase.

Description

[0001] TEC KINASE AGONISTS AND THEIR USE IN ENHANCING LYMPHOCYTE ACTIVATION

[0002] RELATED APPLICATIONS

[0003] This patent application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application U.S.S.N. 63 / 640,561, filed April 30, 2024, and U.S. Provisional Patent Application Number U.S.S.N. 63 / 640,573, filed April 30, 2024, each of which is incorporated herein by reference in its entirety.

[0004] BACKGROUND

[0005] The B cell receptor (BCR) contributes to both the initial burst of B cell proliferation in response to antigens, and for the differentiation of plasma cells and memory B cells during germinal center reactions. However, the BCR’s precise role in germinal center selection has been difficult to assess. Mechanistic investigations into its modulation during the decision of B cell fate suffer from experimental imitations due to the fundamental role of the BCR in B cell biology. Bruton’s tyrosine kinase (BTK), a member of the TEC family, is a key component of BCR signaling.

[0006] Bruton’s tyrosine kinase (BTK) and IL-2 inducible T cell kinase (ITK), members of the Tec kinase family, are particularly active in B and T lymphocytes, respectively. BTK inhibition is an FDA-approved strategy to treat various B cell leukemias and lymphomas as evidenced by several generations of orthosteric BTK inhibitor drugs. There are no modalities that enhance BCR signaling via BTK modulation in humans.

[0007] ITK is an integrator of signals from T cell antigen receptor (TCR) and the costimulatory molecule CD28. There are no ITK inhibitors in clinical use nor are there precedents for ITK agonists, in vitro or in vivo. There is a need for lymphocyte modulation induced therapeutic benefits, particularly to treat serious diseases such as cancers.

[0008] The compounds and methods disclosed herein address these and other needs.

[0009] SUMMARY

[0010] In accordance with the purposes of the disclosed materials and methods, as embodied and broadly described herein, the disclosed subject matter, in one aspect, relates to compounds and methods of using thereof.

[0011] Thus, in some embodiments, provided herein are compounds according to Formula

[0012] I: or a pharmaceutically acceptable salt thereof; wherein X is O, S, or N(H); R1is H or C1-20 alkyl; R2is a halogen; each RAis independently OH, halogen, C1-20 alkyl, C1-20 alkoxy, amino, C1-20 alkylamino, C2-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, or 7-20 membered alkylaryl; n is 0, 1, 2, or 3; R3is H, C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-20 acyl, C1-20 alkoxy, 7-20 membered aryloxy, C1-20 alkylamino, C2-20 dialkylamino, halogen, or amino; and if X is S and R3is -OCHF2, then R2is H, Br, or F.

[0013] In further embodiments, provided herein are compounds according to Formula II: or a pharmaceutically acceptable salt thereof; wherein Z is O, S, or N(H); R6is a halogen; R4is substituted or unsubstituted C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-20 acyl, C1-20 alkoxy, 7-20 membered aryloxy, C1-20 alkylamino, C2-20 dialkylamino, halogen, or amino; and R5is

[0014]

[0015] Additionally, in some embodiments provided herein are compounds according to

[0016] Formula III: (Formula III), or a pharmaceutically acceptable salt thereof; wherein A is O, S, or N(H); R8is H or halogen; R9is C1-20 alkyl, 3-20 membered cycloalkyl, or 6-20 membered aryl; R7ais , then R8is halogen; and (b) when

[0017] In some embodiments, the disclosure provides compounds according to Formula IV: or a pharmaceutically acceptable salt thereof; wherein Further provided herein are methods of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula V : or a pharmaceutically acceptable salt thereof; wherein X is O, S, or N(H); R1is H or substituted or unsubstituted C1-20 alkyl; R2is H or halogen; each RAis independently OH, halogen, C1-20 alkyl, C1-20 alkoxy, amino, C1-20 alkylamino, C2-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, or 7-20 membered alkylaryl; n is 0, 1, 2, or 3; and R3is H, substituted or unsubstituted C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-20 acyl, C1-20 alkoxy, 7-20 membered aryloxy, C1-20 alkylamino, C2-20 dialkylamino, halogen, or amino.

[0018] Also provided herein are methods of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula VI (Formula VI), or a pharmaceutically acceptable salt thereof; wherein Z is O, S, or N(H); R6is a halogen; R4is substituted or unsubstituted C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-20 acyl, C1-20 alkoxy, 7-20 membered aryloxy, C1-20 alkylamino, C2-20 dialkylamino, halogen, or amino; and R5is

[0019]

[0020] Further provided herein are methods of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula VII: (Formula VII) or a pharmaceutically acceptable salt or; wherein A is O, S, or N(H); R8is H or halogen; R9is Ci-20 alkyl, 3-20 membered cycloalkyl, or 6-20 membered aryl; and R7bis with the proviso that when is halogen.

[0021] Also provided herein are methods of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula VIII: (Formula VIII) or a pharmaceutically acceptable salt thereof; wherein

[0022] In further embodiments, methods of reducing solid tumor size are provided comprising administering compounds disclosed herein. In various embodiments, solid tumor size is reduced through lymphocyte activation.

[0023] Additionally, methods of increasing lymphocyte activation are provided, including subjecting the lymphocyte cell surface or intracellular compartments to any of the compounds disclosed herein. In some embodiments, methods of activating TEC kinase are provided, including targeting a surface of the TEC kinase with any of the compounds disclosed herein. In some embodiments, methods of treating cancer are provided, including administering a therapeutically effective amount of a compound according to Formula IA, or a pharmaceutically acceptable salt thereof: (Formula IA), wherein X is S or O; wherein R1is 6-20 membered aryl, 7-20 membered alkylaryl, or 3-20 membered cycloalkyl; R2is H, substituted or unsubstituted C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-C20 acyl, C1-C20 alkoxy, or 7-20 membered aryloxy; each RAis independently OH, halogen, Ci- 20 alkyl, C1-20 alkoxy, amino, C1-20 alkylamino, C2-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, or 7-20 membered arylalkyl, or two RAgroups adjacent to one another together with atoms to which they are attached to form a fused 5 to 7 membered cyclic moiety; and n is 0, 1, or 2.

[0024] In further embodiments, methods of treating cancer are provided, including administering a therapeutically effective amount of a compound according to Formula IIA, or a pharmaceutically acceptable salt thereof: wherein R3is H or acyl; R4and R5are independently H, C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, or 3-20 membered cycloalkyl, or R4and R5, together with the nitrogen atom to which they are attached, form a 5 to 10 membered cyclic moiety; each RAis independently C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl,

[0025] 6-20 membered aryl, 7-20 membered alkylaryl, or 3-20 membered cycloalkyl; and n is 0, 1, or 2. Additionally, methods of treating cancer are provided, including administering a therapeutically effective amount of a compound according to Formula IIIA, or a pharmaceutically acceptable salt thereof: (Formula IIIA), wherein X is S or O; each RAis independently OH, halogen, C1-20 alkyl, C1-20 alkoxy, amino, C1-20 alkylamino, C2-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, 7-20 membered arylalkyl, or two RAgroups adjacent to one another together with atoms to which they are attached to form a fused 5 to 10 membered cyclic moiety; and n is 0, l, or 2.

[0026] In some embodiments, methods of treating cancer are provided, including administering a therapeutically effective amount of a compound according to Formula IVA, or a pharmaceutically acceptable salt thereof: (Formula IVA), wherein R6is H, OH, halogen, C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-C20 acyl, C1-C20 alkoxy, or 7-20 membered aryloxy; each RAis independently OH, halogen, C1-20 alkyl, C1-20 alkoxy, amino, C1-20 alkylamino, C2-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, or 7- 20 membered arylalkyl, or two RAgroups adjacent to one another together with atoms to which they are attached to form a fused 5 to 10 membered cyclic moiety; and n is 0, 1, or 2.

[0027] In further embodiments, methods of reducing solid tumor size are provided, including subjecting animal hosts or human patients bearing tumors to any of the compounds disclosed herein.

[0028] Additionally, methods of increasing lymphocyte activation are provided, including subjecting the lymphocyte to any of the compounds disclosed herein. In some embodiments, methods of activating TEC kinase are provided, including targeting the TEC kinase with any of the compounds disclosed herein.

[0029] Additional advantages will be set forth in part in the description that follows, and in part will be obvious from the description, or may be learned by practice of the aspects described below. The advantages described below will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive.

[0030] BRIEF DESCRIPTION OF THE DRAWINGS

[0031] The accompanying figures, which are incorporated in and constitute a part of this specification, illustrate several aspects described below.

[0032] Figures 1A - ID show a computational docking to BTK narrows the field of small molecules. (A)Virtual and high-throughput screening workflow: The identified allosteric pocket (red) in the autoinhibited structure of BTK was screened against Zinc database and Chemdiv libraries by docking using DOCKBLASTER and GLIDE. Ligands with high docking scores were subjected to standard compound filters to remove compounds with known toxicities. Selected compounds were then experimentally screened for modulation of NUR77 expression in isolated splenic B cells, or by STD-NMR to detect direct binding to BTK. (B) Identification of allosteric sites in BTK: Allosteric pocket (red) identified in the autoinhibited structure of BTK (PDB:4XI2) Schrodinger’s SiteMap is lined by residues from SH3 (yellow), linker and kinase domains (grey). The active site is represented as grey surface with Y551 on the activation loop represented in stick. (C) Distribution of % SAT values for a subset of the compounds in our small molecule library. Saturation Transfer Difference Experiment for small molecules at 200 pM for all compounds and the protein was at lOpM. Distribution of % SAT values for a subset of the compounds in the small molecule library. (D) STD spectra or a positive hit. Saturation is applied to the sample during the “saturation on spectra”. If the compound binds to the protein, the saturation is transferred, and its signal is attenuated. The “saturation off spectra” is the reference spectra. The “difference spectra” is obtained by subtracting the on spectra from the off spectra. Using these values % SAT can be calculated. If signal occurs in the Difference spectra, it is an indication that the compound is binding. Figures 2A - 2D show Nur77 screen reveals compounds that modulate BCR signaling in both inhibitory and enhancing fashions. (A) BCR signaling pathway after BCR triggering by a-IgM and resulting NUR77 expression. (B) Volcano plot of small molecule screen using NUR77-GFP expression in naive B cells 16 hours after BCR engagement and compound treatment. (C) Representative flow cytometric histograms showing NUR77-GFP expression in naive B cells 16 hours post BCR engagement and compound treatment. Each plot shows no stim control (gray), 1 or 5 pg / mL a-IgM control (black) and compound treated (color) samples. (D) Modulation of NUR77-GFP expression by titrated compounds after 16 hours of BCR engagement and compound treatment in naive B cells. BUR77-GFP+ frequency was normalized to vehicle control.

[0033] Figures 3A-3F show compound treatment enhances calcium flux in a BTK dependent manner. (A) Compiled calcium flux of Fluo3 and FuraRed labeled wild type B cells treated for 60 minutes with vehicle (DMSO) or 5 nM of ibrutinib (a known BTK inhibitor, used as a positive control agent). Calcium flux was measured by flow cytometry with the addition of 5 pg / mL a-IgM after 30 seconds of event recording. (B) Area under the curve was measured between 45 and 250 seconds. (C) Compiled calcium flux of FLuo3 and FureRed labeled wild type B cells treated for 60 minutes with vehicle or 50 pM compound (Cl 11). Flux was measured by flow cytometry with the addition of 5 pg / mL of a-IgM at 30 seconds. (D) Area under the curved calculated between 45 and 250 seconds, (e) Compiled calcium flux of FLuo3 and FureRad labeled Cd21creBTKFI / YB cells treated for 60 minutes with vehicle or 25 pM compound. Flux was measured by flow cytometry with the addition of 5 pg / mL of a-IgM at 30 seconds. (F) Area under the curve calculated between 45 and 250 seconds.

[0034] Figures 4A-4F show compound treatment sustains BCR signaling in presence of BTK inhibitor. (A) Experiment layout of NUR77-GFP B cells that were cultured for 16 hours with 50 pM compound and 5 nM BTK active site inhibitor (GDC-0853) then analyzed by flow cytometry. (B) Representative flow cytometry histograms of NUR77-GFP expression after BCR engagement and dual BTK target treatment. (C) Nur77-GFP expression normalized to vehicle (DMSO) control. (D) Experiment layout of NUR77-GFP B cells treated with 50 pM compound, after 4 hours of culture 5 nM BTK active site inhibitor (GDC-0853) was added. Analysis by flow was at 16 hour timepoint. (E) Representative flow cytometry histograms of NUR77-GFP expression after BCR engagement and dual BTK target treatment. (F) NUR77- GFP expression normalized to vehicle (DMSO) control. Figures 5A-5D show compound treatment pushes B cells towards a plasma cell differentiation in vitro. (A) Representative flow cytometric histograms (left) and summarized data (right) showing B cell divisions after 3.5 days in culture with 5 pg / mL aCD40 and algM plus vehicle control (DMSO) or 25 pM compound (Cl 11). (B) Representative flow cytometric plots of B cells cultured for 3.5 days on top of NB21 feeder cells, supplemented with 5 pg / mL algM, and treated with titrated compound, (C) Summarized data of total B cell count and (D) CD138+ frequency after 3.5 days of culture on top of NB21 feeder cells, supplemented with 5 pg / mL algM, and treated with titrated compound.

[0035] Figures 6A-6F show compound treatment leads to B-cell activation in live mouse models, and specifically, that treatment enhances germinal center and drives larger IgGl+ plasma cell fate in vivo. Mice injected I.P. on DO and D4 with sRBC plus vehicle control (DMSO) or 5 mM compound (Cl 11) and B-cells from the spleen were analyzed on D14. (A) Representative flow plots (left), frequency (middle), and absolute number (right) of germinal B cells. (B) Representative IHC images (left), number of GL7+ areas (middle), and area of germinal centers measured using ImageJ (right). (C) Representative flow plots (leftO< frequency (middle), and absolute number of IgGl+ germinal center B cells. (D) Representative flow plots (left), frequency (middle), and absolute number (right) of plasma cells. (E) Representative flow plots (left), frequency (middle), and absolute number (right) of IgGl+ plasma cells. (F) Representative ELISPOT images (and number of IgGl+ spots / 1 x 106 cells (right).

[0036] Figure 7 shows an example of a synthetic route for a compound with the formula of 5-chloro-N-(4-(difluoromethoxy)phenyl)-2-(methylthio)pyrimidine-4-carboxamide (C37).

[0037] Figure 8 shows an example synthetic route for a compound with the formula of N-(4- (cyclopentylthio)phenyl)-3-methylpyrazine-2-carboxamide (Cl 11).

[0038] Figure 9 shows (lH-benzo[d]imidazol-5-yl)(3-((phenylthio)methyl)-2,5-dihydro-lH- pyrrol-l-yl)methanone (C79). Boc represents a protective group.

[0039] DETAILED DESCRIPTION

[0040] Definitions

[0041] As used herein, “comprising” is to be interpreted as specifying the presence of the stated features, integers, steps, or components as referred to, but does not preclude the presence or addition of one or more features, integers, steps, or components, or groups thereof. Moreover, each of the terms “by”, “comprising,” “comprises”, “comprised of,” “including,” “includes,” “included,” “involving,” “involves,” “involved,” and “such as” are used in their open, non-limiting sense and may be used interchangeably. Further, the term “comprising” is intended to include examples and aspects encompassed by the terms “consisting essentially of’ and “consisting of.” Similarly, the term “consisting essentially of’ is intended to include examples encompassed by the term “consisting of.”

[0042] As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a compound”, “a composition”, or “a disorder”, includes, but is not limited to, two or more such compounds, compositions, or disorders, and the like.

[0043] It should be noted that ratios, concentrations, amounts, and other numerical data can be expressed herein in a range format. It can be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as “about” that particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is also disclosed. Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it can be understood that the particular value forms a further aspect. For example, if the value “about 10” is disclosed, then “10” is also disclosed.

[0044] When a range is expressed, a further aspect includes from the one particular value and / or to the other particular value. For example, where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the disclosure, e.g., the phrase “x to y” includes the range from ‘x’ to ‘y’ as well as the range greater than ‘x’ and less than ‘y’. The range can also be expressed as an upper limit, e.g., ‘about x, y, z, or less’ and should be interpreted to include the specific ranges of ‘about x’, ‘about y’, and ‘about z’ as well as the ranges of ‘less than x’, less than y’, and ‘less than z’. Likewise, the phrase ‘about x, y, z, or greater’ should be interpreted to include the specific ranges of ‘about x’, ‘about y’, and ‘about z’ as well as the ranges of ‘greater than x’, greater than y’, and ‘greater than z’. In addition, the phrase “about ‘x’ to ‘y’”, where ‘x’ and ‘y’ are numerical values, includes “about ‘x’ to about ‘y’”.

[0045] It is to be understood that such a range format is used for convenience and brevity, and thus, should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and subrange is explicitly recited. To illustrate, a numerical range of “about 0.1% to 5%” should be interpreted to include not only the explicitly recited values of about 0.1% to about 5%, but also include individual values (e.g., about 1%, about 2%, about 3%, and about 4%) and the sub-ranges (e.g., about 0.5% to about 1.1%; about 5% to about 2.4%; about 0.5% to about 3.2%, and about 0.5% to about 4.4%, and other possible sub-ranges) within the indicated range.

[0046] As used herein, the terms “about,” “approximate,” “at or about,” and “substantially” mean that the amount or value in question can be the exact value or a value that provides equivalent results or effects as recited in the claims or taught herein. That is, it is understood that amounts, sizes, formulations, parameters, and other quantities and characteristics are not and need not be exact but may be approximate and / or larger or smaller, as desired, reflecting tolerances, conversion factors, rounding off, measurement error and the like, and other factors known to those of skill in the art such that equivalent results or effects are obtained. In some circumstances, the value that provides equivalent results or effects cannot be reasonably determined. In such cases, it is generally understood, as used herein, that “about” and “at or about” mean the nominal value indicated ±10% variation unless otherwise indicated or inferred. In general, an amount, size, formulation, parameter or other quantity or characteristic is “about,” “approximate,” or “at or about” whether or not expressly stated to be such. It is understood that where “about,” “approximate,” or “at or about” is used before a quantitative value, the parameter also includes the specific quantitative value itself, unless specifically stated otherwise. As used herein, the term “substantially free,” when used in the context of a composition or component of a composition that is substantially absent, is intended to refer to an amount that is then about 1% by weight or less, e.g., less than about 0.5% by weight, less than about 0.1% by weight, less than about 0.05% by weight, or less than about 0.0 % by weight of the stated material, based on the total weight of the composition.

[0047] The term “subject” preferably refers to a human in need of treatment with an anticancer agent or treatment for any purpose, and more preferably a human in need of such a treatment to treat cancer, or a precancerous condition or lesion. However, the term “patient” can also refer to non-human animals, preferably mammals such as dogs, cats, horses, cows, pigs, sheep and non-human primates, among others, that are in need of treatment with an anti-cancer agent or treatment. The term “treating” refers to the medical management of a patient with the intent to cure, ameliorate, stabilize, or prevent a disease, pathological condition, or disorder. This term includes active treatment, that is, treatment directed specifically toward the improvement of a disease, pathological condition, or disorder, and also includes causal treatment, that is, treatment directed toward removal of the cause of the associated disease, pathological condition, or disorder. In addition, this term includes palliative treatment, that is, treatment designed for the relief of symptoms rather than the curing of the disease, pathological condition, or disorder; preventative treatment, that is, treatment directed to minimizing or partially or completely inhibiting the development of the associated disease, pathological condition, or disorder; and supportive treatment, that is, treatment employed to supplement another specific therapy directed toward the improvement of the associated disease, pathological condition, or disorder.

[0048] The term “therapeutically effective” refers to the amount of the composition used is of sufficient quantity to ameliorate one or more causes or symptoms of a disease or disorder. Such amelioration only requires a reduction or alteration, not necessarily elimination.

[0049] The term “pharmaceutically acceptable” refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0050] The term “carrier” means a compound, composition, substance, or structure that, when in combination with a compound or composition, aids or facilitates preparation, storage, administration, delivery, effectiveness, selectivity, or any other feature of the compound or composition for its intended use or purpose. For example, a carrier can be selected to minimize any degradation of the active ingredient and to minimize any adverse side effects in the subject.

[0051] The term “pharmaceutically acceptable salts” refers to physiologically and pharmaceutically acceptable salts of the compounds of the invention: i.e., salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects thereto. Chemical Definitions

[0052] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0053] The organic moieties mentioned when defining variable positions within the general formulae described herein (e.g., the term “halogen”) are collective terms for the individual substituents encompassed by the organic moiety. The prefix Cn-Cmpreceding a group or moiety indicates, in each case, the possible number of carbon atoms in the group or moiety that follows.

[0054] As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, and aromatic and nonaromatic substituents of organic compounds. Illustrative substituents include, for example, those described below. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this disclosure, the heteroatoms, such as nitrogen, can have hydrogen substituents and / or any permissible substituents of organic compounds described herein which satisfy the valencies of the heteroatoms. This disclosure is not intended to be limited in any manner by the permissible substituents of organic compounds. Also, the terms “substitution” or “substituted with” include the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., a compound that does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc.

[0055] “Z1,” “Z2,” “Z3,” and “Z4” are used herein illustratively as generic symbols to represent various specific substituents. These symbols can be any substituent, not limited to those disclosed herein, and when they are defined to be certain substituents in one instance, they can, in another instance, be defined as some other substituents.

[0056] As used herein, the term “alkyl” refers to saturated, straight-chained or branched saturated hydrocarbon moieties. Unless otherwise specified, C1-C24 (e.g., C1-C22, C1-C20, Ci-Cis, C1-C16, Ci-C 14, C1-C12, C1-C10, Ci-Cs, Ci-Ce, or C1-C4) alkyl groups are intended. Examples of alkyl groups include methyl, ethyl, propyl, 1-methyl-ethyl, butyl, 1 -methylpropyl, 2-methyl-propyl, 1,1-dimethyl-ethyl, pentyl, 1-methyl-butyl, 2-methyl-butyl, 3- methyl-butyl, 2,2-dimethyl-propyl, 1-ethyl-propyl, hexyl, 1,1-dimethyl-propyl, 1,2- dimethyl-propyl, 1-methyl-pentyl, 2-methyl-pentyl, 3-methyl-pentyl, 4-methyl-pentyl, 1,1- dimethyl-butyl, 1,2-dimethyl-butyl, 1,3-dimethyl-butyl, 2,2-dimethyl-butyl, 2,3-dimethyl- butyl, 3,3-dimethyl-butyl, 1-ethyl-butyl, 2-ethyl-butyl, 1,1,2-trimethyl-propyl, 1,2,2- trimethyl-propyl, 1 -ethyl- 1-methyl-propyl, l-ethyl-2-methyl-propyl, heptyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, eicosyl, tetracosyl, and the like. Alkyl substituents may be unsubstituted or substituted with one or more chemical moieties. The alkyl group can be substituted with one or more groups including, but not limited to, hydroxyl, halogen, acetal, acyl, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, cyano, carboxylic acid, ester, ether, carbonate ester, carbamate ester, ketone, nitro, phosphonyl, silyl, sulfo-oxo, sulfonyl, sulfone, sulfoxide, or thiol, as described below, provided that the substituents are sterically compatible and the rules of chemical bonding and strain energy are satisfied.

[0057] Throughout the specification “alkyl” is generally used to refer to both unsubstituted alkyl groups and substituted alkyl groups; however, substituted alkyl groups are also specifically referred to herein by identifying the specific substituent(s) on the alkyl group. For example, the term “halogenated alkyl” or “haloalkyl” specifically refers to an alkyl group that is substituted with one or more halides (halogens; e.g., fluorine, chlorine, bromine, or iodine). The term “alkoxyalkyl” specifically refers to an alkyl group that is substituted with one or more alkoxy groups, as described below. The term “alkylamino” specifically refers to an alkyl group that is substituted with one or more amino groups, as described below, and the like. When “alkyl” is used in one instance and a specific term such as “alkylalcohol” is used in another, it is not meant to imply that the term “alkyl” does not also refer to specific terms such as “alkylalcohol” and the like.

[0058] This practice is also used for other groups described herein. That is, while a term such as “cycloalkyl” refers to both unsubstituted and substituted cycloalkyl moieties, the substituted moieties can, in addition, be specifically identified herein; for example, a particular substituted cycloalkyl can be referred to as, e.g., an “alkylcycloalkyl.” Similarly, a substituted alkoxy can be specifically referred to as, e.g., a “halogenated alkoxy,” a particular substituted alkenyl can be, e.g., an “alkenylalcohol,” and the like. Again, the practice of using a general term, such as “cycloalkyl,” and a specific term, such as “alkylcycloalkyl,” is not meant to imply that the general term does not also include the specific term.

[0059] As used herein, the term “alkenyl” refers to unsaturated, straight-chained, or branched hydrocarbon moieties containing a double bond. Unless otherwise specified, C2- C24 (e.g., C2-C22, C2-C20, C2-C18, C2-C16, C2-C14, C2-C12, C2-C10, C2-C8, C2-C6, or C2-C4) alkenyl groups are intended. Alkenyl groups may contain more than one unsaturated bond. Examples include ethenyl, 1 -propenyl, 2-propenyl, 1 -methylethenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1 -methyl- 1 -propenyl, 2-methyl-l -propenyl, l-methyl-2-propenyl, 2-methyl-2- propenyl, 1 -pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1 -methyl- 1-butenyl, 2-methyl-l- butenyl, 3-methyl- 1-butenyl, l-methyl-2-butenyl, 2-methyl-2-butenyl, 3-methyl-2-butenyl,

[0060] 1-methyl-3-butenyl, 2-methyl-3-butenyl, 3-methyl-3-butenyl, l,l-dimethyl-2-propenyl, 1,2- dimethyl-1 -propenyl, l,2-dimethyl-2-propenyl, 1 -ethyl- 1 -propenyl, l-ethyl-2-propenyl, 1- hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1 -methyl- 1 -pentenyl, 2-methyl-l- pentenyl, 3-methyl- 1 -pentenyl, 4-methyl-l -pentenyl, l-methyl-2-pentenyl, 2-methyl-2- pentenyl, 3-methyl-2-pentenyl, 4-methyl-2-pentenyl, l-methyl-3-pentenyl, 2-methyl-3- pentenyl, 3-methyl-3-pentenyl, 4-methyl-3-pentenyl, l-methyl-4-pentenyl, 2-methyl-4- pentenyl, 3-methyl-4-pentenyl, 4-methyl-4-pentenyl, l,l-dimethyl-2-butenyl, 1,1-dimethyl- 3-butenyl, 1,2-dimethyl- 1-butenyl, l,2-dimethyl-2-butenyl, l,2-dimethyl-3-butenyl, 1,3- dimethyl- 1-butenyl, l,3-dimethyl-2-butenyl, l,3-dimethyl-3-butenyl, 2,2-dimethyl-3- butenyl, 2, 3-dimethyl- 1-butenyl, 2,3-dimethyl-2-butenyl, 2,3-dimethyl-3-butenyl, 3,3- dimethyl- 1-butenyl, 3,3-dimethyl-2-butenyl, 1 -ethyl- 1-butenyl, l-ethyl-2-butenyl, 1-ethyl- 3-butenyl, 2-ethyl- 1-butenyl, 2-ethyl-2-butenyl, 2-ethyl-3-butenyl, l,l,2-trimethyl-2- propenyl, 1 -ethyl- l-methyl-2-propenyl, l-ethyl-2-methyl-l -propenyl, and l-ethyl-2-methyl-

[0061] 2-propenyl. The term “vinyl” refers to a group having the structure -CH=CH2; 1 -propenyl refers to a group with the structure -CH=CH-CH3; and 2-propenyl refers to a group with the structure -CH2-CH=CH2. Asymmetric structures such as (Z1Z2)C=C(Z3Z4) are intended to include both the E and Z isomers. This can be presumed in structural formulae herein wherein an asymmetric alkene is present, or it can be explicitly indicated by the bond symbol C=C. Alkenyl substituents may be unsubstituted or substituted with one or more chemical moieties. Examples of suitable substituents include, for example, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, acetal, acyl, aldehyde, amino, cyano, carboxylic acid, ester, ether, carbonate ester, carbamate ester, halide, hydroxyl, ketone, nitro, phosphonyl, silyl, sulfo-oxo, sulfonyl, sulfone, sulfoxide, or thiol, as described below, provided that the substituents are sterically compatible and the rules of chemical bonding and strain energy are satisfied. Examples of “alkenyl” also include cycloalkenyl.

[0062] As used herein, the term “alkynyl” represents straight-chained or branched hydrocarbon moieties containing a triple bond. Unless otherwise specified, C2-C24 (e.g., C2- C24, C2-C20, C2-C18, C2-C16, C2-C14, C2-C12, C2-C10, C2-C8, C2-C6, or C2-C4) alkynyl groups are intended. Alkynyl groups may contain more than one unsaturated bond. Examples include C2-Ce-alkynyl, such as ethynyl, 1-propynyl, 2-propynyl (or propargyl), 1-butynyl,

[0063] 2-butynyl, 3-butynyl, l-methyl-2-propynyl, 1 -pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl,

[0064] 3-methyl- 1-butynyl, l-methyl-2-butynyl, l-methyl-3-butynyl, 2-methyl-3-butynyl, 1,1- dimethyl-2-propynyl, l-ethyl-2-propynyl, 1 -hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5- hexynyl, 3-methyl- 1 -pentynyl, 4-methyl-l -pentynyl, l-methyl-2-pentynyl, 4-methyl-2- pentynyl, l-methyl-3-pentynyl, 2-methyl-3-pentynyl, l-methyl-4-pentynyl, 2-methyl-4- pentynyl, 3-methyl-4-pentynyl, l,l-dimethyl-2-butynyl, l,l-dimethyl-3-butynyl, 1,2- dimethyl-3-butynyl, 2,2-dimethyl-3-butynyl, 3,3-dimethyl-l-butynyl, l-ethyl-2-butynyl, 1- ethyl-3-butynyl, 2-ethyl-3-butynyl, and 1 -ethyl- l-methyl-2-propynyl. Alkynyl substituents may be unsubstituted or substituted with one or more chemical moieties. Examples of suitable substituents include, for example, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, acetal, acyl, aldehyde, amino, cyano, carboxylic acid, ester, ether, carbonate ester, carbamate ester, halide, hydroxyl, ketone, nitro, phosphonyl, silyl, sulfo-oxo, sulfonyl, sulfone, sulfoxide, or thiol, as described below. Examples of “alkynyl” also include cycloalkynyl.

[0065] As used herein, the term “aryl,” as well as derivative terms such as aryloxy, refers to groups that include a monovalent aromatic carbocyclic group of from 3 to 50 carbon atoms. Aryl groups can include a single ring or multiple condensed rings. In some embodiments, aryl groups include Ce-Cio aryl groups. Examples of aryl groups include, but are not limited to, benzene, phenyl, biphenyl, naphthyl, tetrahydronaphthyl, phenylcyclopropyl, phenoxybenzene, and indanyl. The term “aryl” also includes “heteroaryl,” which is defined as a group that contains an aromatic group that has at least one heteroatom incorporated within the ring of the aromatic group. Examples of heteroatoms include, but are not limited to, nitrogen, oxygen, sulfur, and phosphorus. The term “non-heteroaryl,” which is also included in the term “aryl,” defines a group that contains an aromatic group that does not contain a heteroatom. The aryl substituents may be unsubstituted or substituted with one or more chemical moieties. Examples of suitable substituents include, for example, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, acetal, acyl, aldehyde, amino, cyano, carboxylic acid, ester, ether, carbonate ester, carbamate ester, halide, hydroxyl, ketone, nitro, phosphonyl, silyl, sulfo-oxo, sulfonyl, sulfone, sulfoxide, or thiol as described herein. The term “biaryl” is a specific type of aryl group and is included in the definition of aryl. Biaryl refers to two aryl groups that are bound together via a fused ring structure, as in naphthalene, or are attached via one or more carbon-carbon bonds, as in biphenyl.

[0066] The term “cycloalkyl” as used herein is a non-aromatic carbon-based ring composed of at least three carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc. The term “heterocycloalkyl” is a cycloalkyl group as defined above where at least one of the carbon atoms of the ring is substituted with a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus. The cycloalkyl group and heterocycloalkyl group can be substituted or unsubstituted. The cycloalkyl group and heterocyclo alkyl group can be substituted with one or more groups including, but not limited to, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, acetal, acyl, aldehyde, amino, cyano, carboxylic acid, ester, ether, carbonate ester, carbamate ester, halide, hydroxyl, ketone, nitro, phosphonyl, silyl, sulfo-oxo, sulfonyl, sulfone, sulfoxide, or thiol as described herein.

[0067] The term “cycloalkenyl” as used herein is a non-aromatic carbon-based ring composed of at least three carbon atoms and containing at least one double bound, i.e., C=C. Examples of cycloalkenyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, and the like. The term “heterocycloalkenyl” is a type of cycloalkenyl group as defined above and is included within the meaning of the term “cycloalkenyl,” where at least one of the carbon atoms of the ring is substituted with a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus. The cycloalkenyl group and heterocyclo alkenyl group can be substituted or unsubstituted. The cycloalkenyl group and heterocycloalkenyl group can be substituted with one or more groups including, but not limited to, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, acetal, acyl, aldehyde, amino, cyano, carboxylic acid, ester, ether, carbonate ester, carbamate ester, halide, hydroxyl, ketone, nitro, phosphonyl, silyl, sulfooxo, sulfonyl, sulfone, sulfoxide, or thiol as described herein.

[0068] The term “cyclic group” is used herein to refer to either aryl groups, non-aryl groups (i.e., cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl groups), or both. Cyclic groups have one or more ring systems (e.g., monocyclic, bicyclic, tricyclic, polycyclic, etc.) that can be substituted or unsubstituted. A cyclic group can contain one or more aryl groups, one or more non-aryl groups, or one or more aryl groups and one or more non-aryl groups.

[0069] The term “acyl” as used herein is represented by the formula -C(O)Z' where Z1can be a hydrogen, hydroxyl, alkoxy, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl group described above. As used herein, the term “acyl” can be used interchangeably with “carbonyl.” Throughout this specification “C(O)” or “CO” is a shorthand notation for C=O. As used herein, the term “alkoxy” as used herein is an alkyl group bound through a single, terminal ether linkage; that is, an “alkoxy” group can be defined as to a group of the formula Z'-O-, where Z1is unsubstituted or substituted alkyl as defined above. Unless otherwise specified, alkoxy groups wherein Z1is a C1-C24 (e.g., C1-C22, C1-C20, Ci-Cis, Ci- Ci6, C1-C14, C1-C12, C1-C10, Ci-Cs, Ci-Ce, or C1-C4) alkyl group are intended. Examples include methoxy, ethoxy, propoxy, 1 -methyl-ethoxy, butoxy, 1-methyl-propoxy, 2-methyl- propoxy, 1,1 -dimethyl-ethoxy, pentoxy, 1-methyl-butyloxy, 2-methyl-butoxy, 3-methyl- butoxy, 2,2-di-methyl-propoxy, 1-ethyl-propoxy, hexoxy, 1,1-dimethyl-propoxy, 1,2- dimethyl-propoxy, 1-methyl-pentoxy, 2-methyl-pentoxy, 3-methyl-pentoxy, 4-methyl- penoxy, 1,1-dimethyl-butoxy, 1,2-dimethyl-butoxy, 1,3-dimethyl-butoxy, 2,2-dimethyl- butoxy, 2,3-dimethyl-butoxy, 3,3-dimethyl-butoxy, 1-ethyl-butoxy, 2-ethylbutoxy, 1,1,2- trimethyl-propoxy, 1,2,2-trimethyl-propoxy, 1 -ethyl- 1-methyl-propoxy, and l-ethyl-2- methyl-propoxy.

[0070] As used herein, the term “aryloxy”, is an aryl group bound through a single, terminal ether linkage, that is, an “aryloxy” group can be defined as a group of formula Z'-O-, wherein Z1is unsubstituted or substituted aryl as defined above. By way of illustration, suitable examples of aryloxy groups include phenoxy and naphthoxy.

[0071] The terms “amine” or “amino” as used herein are represented by the formula — NZ*Z2Z3, where Z1, Z2, and Z3can each be substitution group as described herein, such as hydrogen, an alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl group described above. By way of illustration, the term “alkylamino” refers to an amino group substituted with one or more alkyl groups.

[0072] The terms “amide” or “amido” as used herein are represented by the formula — C(O)NZ1Z2, where Z1and Z2can each be substitution group as described herein, such as hydrogen, an alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl group described above.

[0073] The term “halide” or “halogen” or “halo” as used herein refers to fluorine, chlorine, bromine, and iodine.

[0074] The term “hydroxyl” as used herein is represented by the formula — OH.

[0075] The term “R1,” “R2,” “R3,” “Rn,” etc., where n is some integer, as used herein can, independently, possess one or more of the groups listed above. For example, if R1is a straight chain alkyl group, one of the hydrogen atoms of the alkyl group can optionally be substituted with a hydroxyl group, an alkoxy group, an amine group, an alkyl group, a halide, and the like. Depending upon the groups that are selected, a first group can be incorporated within second group or, alternatively, the first group can be pendant (i.e., attached) to the second group. For example, with the phrase “an alkyl group comprising an amino group,” the amino group can be incorporated within the backbone of the alkyl group. Alternatively, the amino group can be attached to the backbone of the alkyl group. The nature of the group(s) that is (are) selected will determine if the first group is embedded or attached to the second group.

[0076] Unless stated to the contrary, a formula with chemical bonds shown only as solid lines and not as wedges or dashed lines contemplates each possible stereoisomer or mixture of stereoisomer (e.g., each enantiomer, each diastereomer, each meso compound, a racemic mixture, or scalemic mixture).

[0077] A. Compounds and Methods of Use

[0078] Compounds of Formula I

[0079] In some embodiments, the disclosure provides compounds according to Formula I: (Formula I) or a pharmaceutically acceptable salt thereof; wherein X is O, S, or N(H); R1is H or C1-20 alkyl; R2is a halogen; each RAis independently OH, halogen, C1-20 alkyl, C1-20 alkoxy, amino, C1-20 alkylamino, C2-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, or 7-20 membered alkylaryl; n is 0, 1, 2, 3; R3is H, C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-20 acyl, C1-20 alkoxy, 7-20 membered aryloxy, C1-20 alkylamino, C2-20 dialkylamino, halogen, or amino; and if X is S and R3is -OCHF2, then R2is H, Br, or F.

[0080] In some embodiments, R1is CH3 and X is S. In further examples, R1is CH3 and X is

[0081] In certain embodiments, R2is Cl.

[0082] In some embodiments, n is 0.

[0083] In some embodiments, R3is -OCHF2.

[0084] In further embodiments, at least one RAis methyl or both RAare methyl, and n is 2. In some embodiments, R3is -CO2H and R1is not methyl.

[0085] In some examples, R3is OH and RAis not -OCH3.

[0086] Compounds of Formula

[0087] Further provided herein are compounds according to Formula II: (Formula II) or a pharmaceutically acceptable salt thereof; wherein Z is O, S, or N(H); R6is a halogen; R4is substituted or unsubstituted C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-20 acyl, C1-20 alkoxy, 7-20 membered aryloxy, C1-20 alkylamino, C2-20 dialkylamino, halogen, or amino; and R5is

[0088] In some embodiments, R6is Cl.

[0089] In further embodiments, R4is CHF2.

[0090] In certain embodiments,

[0091] In some embodiments, R4is C(O)CH3.

[0092] Compounds of Formula III

[0093] Also provided herein are compounds according to Formula III:

[0094] (Formula III) or a pharmaceutically acceptable salt thereof; wherein A is O, S, or N(H); R8is H or halogen; R9is C1-20 alkyl, 3-20 membered cycloalkyl, or 6-20 membered aryl; R7ais

[0095] In some embodiments, A is S and R9is pentyl. In some embodiments, A is O and R9is methyl.

[0096] Compounds of Formula IV

[0097] Also provided herein, are compounds according to Formula IV : (Formula IV), or a pharmaceutically acceptable salt thereof; wherein

[0098] In some embodiments, n is 0 or 1.

[0099] In further embodiments,

[0100] In certain embodiments, M is S and R11is phenyl.

[0101] In some embodiments, M is O and R11is phenyl.

[0102] In some embodiments,

[0103] In further embodiments, R11is phenyl.

[0104] In certain embodiments,

[0105] In some embodiments, R11is phenyl.

[0106] In some embodiments, n is 1.

[0107] In some embodiments, provided herein is a compound according to one of the following formulae:

[0108]

[0109] Methods of Treating Cancer

[0110] Further provided herein is a method of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula V : (Formula V), or a pharmaceutically acceptable salt thereof; wherein X is O, S, or N(H); R1is H or substituted or unsubstituted C1-20 alkyl; R2is H or halogen; each RAis independently OH, halogen, C1-20 alkyl, C1-20 alkoxy, amino, C1-20 alkylamino, C2-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, or 7-20 membered alkylaryl; n is 0, 1, 2, or 3; R3is H, substituted or unsubstituted C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-20 acyl, C1-20 alkoxy, 7-20 membered aryloxy, C1-20 alkylamino, C2-20 dialkylamino, halogen, or amino.

[0111] In some embodiments, R1is CH3 and X is S. In further embodiments, R1is CH3 and X is O. In some embodiments, R3is -OCHF2. In some embodiments, R2is Cl. In some embodiments, n is 0. In further embodiments, at least one RAis methyl or both RAare methyl and n is 2. In some embodiments, R3is -CO2H and R1is not methyl. In some embodiments, R3is OH and RAis not -OCH3.

[0112] Also provided herein are methods of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula VI: (Formula VI). or a pharmaceutically acceptable salt thereof; wherein Z is O, S, or N(H); R6is a halogen; R4is substituted or unsubstituted C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-20 acyl, C1-20 alkoxy, 7-20 membered aryloxy, C1-20 alkylamino, C2-20 dialkylamino, halogen, or amino; and R5is

[0113]

[0114] In some embodiments, R6is Cl. In further embodiments, R4is -CHF2. In certain embodiments, R4is -C(O)CH3.

[0115] Further provided herein are methods of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula VII: (Formula VII) or a pharmaceutically acceptable salt thereof; wherein A is O, S, or N(H); R8is H or halogen; R9is C1-20 alkyl, 3-20 membered cycloalkyl, or 6-20 membered aryl; R7ais as with the proviso that when is halogen.

[0116] In some embodiments, A is S and R9is pentyl.

[0117] In further embodiments, A is O and R9is methyl.

[0118] Also provided herein is a method of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula VIII: (Formula VIII), or a pharmaceutically acceptable salt thereof; wherein

[0119]

[0120] In some embodiments, n is 0 or 1.

[0121] In further embodiments, In certain embodiments, M is S and R11is phenyl. In some embodiments, M is O and R11is phenyl. In some embodiments, certain embodiments,

[0122] In further embodiments, R11is phenyl.

[0123] In some embodiments, n is 1. In some embodiments, the methods disclosed herein comprises a compound according to one of the following formulae:

[0124]

[0125] B. Methods of Use - Compounds of Formula IA-IVA

[0126] Methods of Treating Cancer

[0127] The present disclosure, in one aspect, provides for methods of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula IA, or a pharmaceutically acceptable salt thereof: (Formula IA), wherein

[0128] X is S or O;

[0129] R1is 6-20 membered aryl, 7-20 membered alkylaryl, or 3-20 membered cycloalkyl;

[0130] R2is H, substituted or unsubstituted C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-C20 acyl, C1-C20 alkoxy, or 7-20 membered aryloxy; each RAis independently OH, halogen, C1-20 alkyl, C1-20 alkoxy, amino, C1-20 alkylamino, C2-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, or 7-20 membered arylalkyl, or two RAgroups adjacent to one another together with atoms to which they are attached to form a fused 5 to 7 membered cyclic moiety; and n is 0, 1, or 2.

[0131] In some embodiments, R1is a 3 to 7 membered aryl optionally substituted with from 1 to 3 RAgroups.

[0132] In some embodiments, R1comprises:

[0133] In some embodiments, R2is a C1-4 alkyl optionally substituted with from 1 to 3 RAgroups. In further embodiments, R2is methyl, difluoromethyl, trifluoromethyl, or fluoromethyl.

[0134] In certain embodiments, R2is a cycloalkyl optionally substituted with from 1 to 3 RAgroups.

[0135] In some embodiments, pentyl.

[0136] In some embodiments, methyl.

[0137] In further embodiments,

[0138] -CHF2.

[0139] In certain embodiments, methyl.

[0140] In some embodiments, and R2is ethyl.

[0141] In some embodiments, is methyl.

[0142] In further embodiments, methyl.

[0143] In certain embodiments, methyl.

[0144] In some embodiments,

[0145] X is O, and R2is methyl. Also provided herein, is a method of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula IIA, or a pharmaceutically acceptable salt thereof: wherein

[0146] R3is H or acyl;

[0147] R4and R5are independently H, C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, or 3-20 membered cycloalkyl, or R4and R5together with the nitrogen atom to which they are attached, form a 5 to 10 membered cyclic moiety; each RAis independently C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, or 3-20 membered cycloalkyl; and n is 0, 1, or 2.

[0148] In some embodiments, R4and R5together with the nitrogen atom to with they are attached form a 9 membered cyclic moiety.

[0149] In further embodiments, at least one of R4and R5comprises RA.

[0150] In certain embodiments, the 9 membered cyclic moiety formed from R4and R5and RAare adjacent to one another and together with atoms to which they are attached form a fused 13 membered cyclic moiety.

[0151]

[0152] In some embodiments, R3comprises an alkylacyl or a cycloalkyl acyl. In further embodiments, (or a deprotonated version thereof, i.e., CCh')-

[0153] In some embodiments, R4and R5together with N in Formula IIA to which they are attached form the two RAmoieties are methyl and carbonyl, and R3is H. Further provided herein is a method of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula IIIA, or a pharmaceutically acceptable salt thereof: (Formula IIIA), wherein

[0154] X is S or O; each RAis independently OH, halogen, C1-20 alkyl, C1-20 alkoxy, amino, C1-20 alkylamino, C2-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, or 7-20 membered arylalkyl, or two RAgroups adjacent to one another together with atoms to which they are attached to form a fused 5 to 10 membered cyclic moiety; and n is 0, 1, or 2. Also provided herein, is a method of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula IVA, or a pharmaceutically acceptable salt thereof: (Formula IVA), wherein

[0155] R6is H, OH, halogen, Ci -20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-C20 acyl, C1-C20 alkoxy, or 7-20 membered aryloxy; each RAis independently OH, halogen, C1-20 alkyl, C1-20 alkoxy, amino, C1-20 alkylamino, C2-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, or 7-20 membered arylalkyl, or two RAgroups adjacent to one another together with atoms to which they are attached to form a fused 5 to 10 membered cyclic moiety; and n is 0, 1, or 2.

[0156] In some embodiments, R6is substituted with one or more RA. In some embodiments, R6is a cycloalkyl, aryl, or alkylaryl and substituted with two instances of RA, which are joined together with the atoms to which they are attached to form a fused 5 to 10 membered cyclic moiety.

[0157] In certain embodiments, RAis C1-6 alkyl. In some embodiments, n is 1 and RAis methyl.

[0158] In some embodiments, n is 1, RAis methyl,

[0159] Methods of Treating Cancer with any of the Compounds Disclosed Herein

[0160] The compounds disclosed herein can be useful as cancer therapeutics.

[0161] In some embodiments, cancer comprises lung cancer, prostate cancer, colon cancer, breast cancer, kidney cancer, or head and neck cancer.

[0162] In further embodiments, lung cancer comprises, but is not limited to, small cell lung cancer (SCLC), non-small cell lung cancer (MSCLC), or carcinoid.

[0163] In certain embodiments, prostate cancer comprises, but is not limited to, adenocarcinoma of the prostate, transitional cell carcinoma of the prostate, squamous cell carcinoma of the prostate, or small cell prostate cancer.

[0164] In some embodiments, colon cancer comprises, but is not limited to, adenocarcinoma of the colon, carcinoid tumors, gastrointestinal stromal tumors in the colon, or lymphoma in the colon.

[0165] In some embodiments, breast cancer comprises, but is not limited to, ductal carcinoma in situ, invasive ductal carcinoma, invasive lobular carcinoma, triple negative breast cancer, inflammatory breast cancer, Paget disease of the breast, angiosarcoma, or Phyllodes tumor.

[0166] In further embodiments, kidney cancer comprises, but is not limited to, renal cell carcinoma or non-renal cell carcinoma. In certain embodiments, renal cell carcinoma comprises, but is not limited to, clear cell carcinoma, papillary carcinoma, chromophobe carcinoma, clear cell papillary carcinoma, collecting duct carcinoma, or medullary carcinoma. In some embodiments, non-renal cell carcinoma comprises, but is not limited to, urothelial carcinoma, Wilms tumor, or renal sarcoma.

[0167] In certain embodiments, head and neck cancer comprises, but is not limited to, hypopharyngeal cancer, laryngeal cancer, lip and oral cavity cancer, metastatic squamous neck cancer with occult primary, nasopharyngeal cancer, oropharyngeal cancer, paranasal sinus and nasal cavity cancer, or salivary gland cancer.

[0168] In further embodiments, cancer comprises Stage I, Stage II, Stage III, or Stage IV cancer.

[0169] Stage I refers to cancer that is small and has not spread beyond the point of origin. Stage II refers to cancer that has grown in size but has not spread beyond the point of origin. Stage III refers to cancer that is larger and may have spread to surrounding tissues, lymph nodes, and / or glands. Stage IV refers to cancer that has spread from beyond the point of origin to at least one other body organ.

[0170] In some embodiments, the methods disclosed herein further comprise administering a second therapeutic agent, e.g., a targeted therapy for the cancer. In certain embodiments, the targeted therapy comprises a small molecule drug, a monoclonal antibody, or any combination thereof.

[0171] In some embodiments, small molecule drug is an inhibitor, such as c-Met inhibitor, HER2 inhibitor, RET inhibitor, EGFR inhibitor, FGFR inhibitor, BTK inhibitor, TRK inhibitor, FET3 inhibitor, JAK2 inhibitor, PARP inhibitor, or AEK inhibitor.

[0172] Examples of small molecule drugs contemplated in the disclosed methods include, but are not limited to, imatinib, gefitinib, bortezomib, erlotinib, sorafenib, sunitinib, dasatinib, vorinostat, nilotinib, lapatinib, temsirolimus, pazopanib, everolimus, romidepsin, icotinib, crizotinib, vandetanib, ruxolitinib, vemurafenib, axitinib, radotinib, bosutinib, vismodegib, carfilzomib, regorafenib, cabozantinib, ponatinib, afatinib, ibrutinib, trametinib, dabrafenib, ceritinib, apatinib, belinostat, Olaparib, idelalisib, alectinib, cobimetinib, palbociclib, Osimertinib, sonidegib, sirolimus, Panobinostat, tucidinostat, ixazomib, Eenvatinib, nintedanib, venetoclax, rucaparib, brigatinib, tivozanib, acalabrutinib, ribociclib, abemaciclib, neratinib, midostaurin, eenasidenib, niraparib, copanlisib, anlotinib, lorlatinib, fruquintinib, binimetinib, encorafenib, dacomitinib, gilteritinib, glasdegib, ivosidenib, Larotrectinib, talazoparib, duvelisib, pexidartinib, Zanubrutinib, entrectinib, erdafitinib, quizartinib, fedratinib, alpelisib, pemigetinib, avapritinib, ripretinib, selumetinib, capmatinib, tepotinib, tucatinib, almonertinib, tazemetostate, selpercatinib, pralsetinib, and any combination thereof.

[0173] In further embodiments, a monoclonal antibody functions in different ways including flagging cancer cells, triggering cell-membrane destruction, blocking cell growth, preventing blood vessel growth, blocking immune system inhibitors, directly attacking cancer cells, delivering radiation treatment, delivering chemotherapy, binding cancer and immune cells, or any combination thereof.

[0174] Examples of a monoclonal antibody contemplated in the disclosed methods include, but are not limited to, trastuzumab, pertuzumab, bevacizumab, rituximab, atezolizumab, avelumab, cemiplimab, cetuximab, daratumumab, dinutuximab, durvalumab, elotuzumab, ipilimumab, isatuximab, mogamulizumab, necitumumab, nivolumab, Obinutuzumab, ofatumumab, olaratumab, panitumumab, pembrolizumab, ramucirumab, and any combination thereof.

[0175] In some embodiments, the methods disclosed herein further comprise administering chemotherapy, radiation therapy, stem cell transplantation, or any combination thereof.

[0176] Chemotherapy is a cancer treatment that utilizes drugs to stop the growth of cancer cells, whether that is by killing the cells or by stopping them from dividing. Chemotherapy can be taken by the mouth or injected into a vein or muscle, in which case the drugs enter the bloodstream and can reach cancer cells throughout the body. In some embodiments chemotherapy can be placed directly into the cerebrospinal fluid, an organ, or a body cavity, in which the drugs mainly impact the cancer cells in those areas. In further embodiments, intrathecal chemotherapy can be used to treat adult AML that spread to the brain and spinal cord. Combination chemotherapy is another option in which treatment utilizes more than one anticancer drug. The way in which chemotherapy is administered depends on the subtype of AML being treated and whether leukemia cells have spread to the brain and spinal cord.

[0177] Radiation therapy is a cancer treatment that utilizes high-energy x-rays or other types of radiation to kill cancer cells or keep them from growing. External radiation therapy uses a machine outside the body to send radiation toward the area of the body with cancer. Total body irradiation is another type of radiation therapy wherein radiation is sent toward the whole body.

[0178] Stem cell transplantation is a treatment in which stem cells (immature blood cells) are removed from the blood or bone marrow of the patient, or donor, and frozen and sorted. After the patient completes chemotherapy and / or radiation, the stored stem cells are thawed and given back to the patient through an infusion. The reinfused stem cells then grow into (and restore) the body’s blood cells. Methods of Reducing Tumor Size

[0179] Also provided herein is a method of reducing solid tumor size comprising subjecting the tumor to any of the compounds disclosed herein.

[0180] In some embodiments, the solid tumor comprises cancer cells, cancer stem cells, connective-tissue cells, immune cells, or any combination thereof.

[0181] In further embodiments, the solid tumor is a carcinoma, a sarcoma, a lymphoma, a blastoma, a melanoma, a germ cell tumor, or a carcinosarcoma.

[0182] Carcinoma refers to cancer that forms in epithelial tissue, wherein epithelial tissue lines most organs, internal passageways in the body, and the skin. Carcinomas can affect the skin, breast, kidney, liver, lungs, pancreas, prostate gland, and head and neck.

[0183] Sarcoma refers to cancer that develops in the bones and soft tissues, including fat, muscles, blood vessels, nerves, deep skin tissues, and fibrous tissues.

[0184] Lymphoma refers to cancer that begins in the cells of the lymph system.

[0185] Blastoma refers to cancer that is caused by malignancies in precursor cells, also called blasts. This type of cancer is more common in children.

[0186] Melanoma refers to a skin cancer that starts in the melanocytes, wherein the melanocytes are cells that make the pigment that gives skin its color.

[0187] Germ cell tumor refers to growths of cells that form from germ cells, which are embryonic cells that develop into reproductive organs. They most often occur in the testicles or ovaries but can occur in other areas such as the abdomen, brain, or chest.

[0188] Carcinosarcoma refers to highly aggressive malignant tumors comprising a mixture of carcinoma and sarcoma.

[0189] In certain embodiments, the solid tumor is a result of breast cancer, lung cancer, prostate cancer, colon cancer, or head and neck cancer.

[0190] In some embodiments, the method further comprises administering chemotherapy, radiation therapy, stem cell transplantation, or any combination thereof.

[0191] The compounds disclosed herein directly enhance BCR signals inside B cells, or TCR and CD28 signals inside T cells to generate fit cells for killing tumors. Methods of Increasing Lymphocyte Activation

[0192] The compounds disclosed herein directly enhance BCR signals in B-cells, or TCR and CD28 signaling in T-cells, thereby endowing on the lymphocytes activated with a compound as disclosed herein molecular properties of cancer-killing lymphocytes.

[0193] As such, also provided herein are methods of increasing lymphocyte activation comprising subjecting lymphocyte cells to any of the compounds disclosed herein.

[0194] In some embodiments, the lymphocyte is a B lymphocyte. In further embodiments, the lymphocyte is a T lymphocyte. A lymphocyte is a type of white blood cell that is part of the immune system. There are two main types of lymphocytes: B cells and T cells. B cells produce antibodies in response to antigens detected within a subject. T cells help B cells make antibodies (CD4 antigen expressing helper subsets, CD4+ Th), make cytokines and cytotoxic granules to activate the immune system and kill target cells, respectively, produce growth factors and cytokines to fortify the epithelial barrier cells in the mucocutaneous tissues and to maintain tissue homeostasis.

[0195] As used herein, lymphocyte activation refers to B and T cell activation in lymphocytes.

[0196] The compounds herein enhance BCR signaling and antibody production. The compounds herein enhance CD28 and TCR mediated signal transduction inside the cells when the T cells see tumor specific moieties (antigens) or foreign antigens derived from pathogens via their TCR, which is the primary activation receptor on T cells.

[0197] Methods of Activating TEC Kinase

[0198] Further provided herein, a method of activating TEC kinase comprising targeting the TEC kinase with any of the compounds disclosed herein.

[0199] TEC kinase family refers to proteins with domain organization including a COOH- terminal kinase domain, preceded by Src homology-2 and 3 (SH2, SH3, respectively) protein interaction domains, a Tec homology domain that includes one or two proline-rich regions that interact intramolecularly or intermolecularly with SH3 domains and contribute to kinase regulation, and in some members, a Plekstrin homology (PH) domain that is essential for membrane localization.

[0200] In some embodiments, the TEC kinase family comprises Bruton’s tyrosine kinase (BTK) or IL-2 inducible kinase (ITK). BTK is required for B cell antigen receptor (BCR) signaling that is required for B cell activation and antibody production. BTK is a non-receptor kinase that can play a pathogenic role in oncogenic signaling, which contributes to the proliferation and survival of leukemic cells in varying B cell malignancies.

[0201] BTK, a member of the TEC family, is a component of BCR signaling. The molecular probes disclosed herein alter B cell activation strength, kinetics, and fate only when the BCR is engaged. BTK agonism strongly favors B cell differentiation into switched plasma cells. BTK agonism, either at the time of the immunization or at the peak of germinal center formation, enhanced humoral immunity, providing for the first time that transient, augmented BCR signaling during germinal center reaction has a direct impact on B cell differentiation ability. In addition to being powerful tools for basic B cell biology investigation, allosteric agonists of BTK can be developed into antigen-licensed B cell adjuvants for vaccines in at risk populations, as well as into cancer immunotherapy drugs which hyperactive B cells in tumor microenvironments during checkpoint inhibitor blockade therapies.

[0202] In some embodiments, the TEC kinase comprises IL-2 inducible T cell kinase (ITK).

[0203] ITK is chromosomally localized at the 5ql-32 position in humans and is a 72 kDa kinase expressed in T cells, NK cells, and mast cells. ITK is expressed at the highest level in naive T cells and thymocytes. ITK is an integrator of signals from TCR and the costimulatory molecule CD28, the two T cell surface proteins utilized to activate naive T cells. ITK is a protein encoded by the ITK gene and contains the following domains: Pleckstrin homology (PH), Tec homology (TH), Src homology 3 (SH3), Src homology 2 (SH2), and Src homology 1 (SHI).

[0204] ITK is an integrator of signals from T cell antigen receptor (TCR) and the costimulatory molecule CD28, the two pivotal T cell surface proteins absolutely required to activate naive T cells. A pathogenic role for ITK was initially thought to involve promotion of the development of T cells involved in allergic and asthmatic diseases. First generation active site covalent inhibitors were clinically ineffective in treating the autoinflammatory conditions. Scientific rationale for the selection of asthma as a disease in which ITK plays a pivotal role has eroded with the emergence of somewhat contrasting studies that have pointed to a broadly essential function of ITK in T cell activation and hence, the maintenance of robust and healthy immune functions. Recent focus has switched to testing ITK inhibition in tissue- specific inflammatory disorders (e.g. colitis) and specific T cell tumor types. Early clinical trials are on-going and presently, there are no ITK inhibitors in clinical use nor are there precedents for ITK agonists, in vitro or in vivo.

[0205] Small molecule allosteric agonists described herein are first-in-class modulators of BTK or ITK. Utility of these novel compounds for treating cancer and boosting immune responses to foreign proteins is addressed.

[0206] In some embodiments, the compound used in any of the methods disclosed herein comprises one of the following formulae: combination thereof.

[0207] Also provided herein is a method comprising administering a compound of the disclosure (e.g., a compound of Formula IA-IVA, or I- VIII), or a pharmaceutically acceptable salt thereof, to a subject.

[0208] A number of embodiments of the disclosure have been described. Nevertheless, it will be understood that various modifications may be made without departing from the spirit and scope of the invention. Accordingly, other embodiments are within the scope of the following claims.

[0209] By way of non-limiting illustration, examples of certain embodiments of the present disclosure are given below.

[0210] EXAMPLES

[0211] The following examples are set forth below to illustrate the methods and results according to the disclosed subject matter. These examples are not intended to be inclusive of all aspects of the subject matter disclosed herein, but rather to illustrate representative methods and results. These examples are not intended to exclude equivalents and variations of the present invention, which are apparent to one skilled in the art.

[0212] Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.), but some errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in °C or is at ambient temperature, and pressure is at or near atmospheric. There are numerous variations and combinations of reaction conditions, e.g., component concentrations, temperatures, pressures, and other reaction ranges and conditions that can be used to optimize the product purity and yield obtained from the described process. Only reasonable and routine experimentation will be required to optimize such process conditions. Example 1: Tunable B cell response through allosteric modulation of Bruton’s tyrosine kinase

[0213] The B cell receptor (BCR) is essential for both the initial burst of B cell proliferation in response to antigens, and for the differentiation of plasma cells and memory B cells during germinal center reactions. However, the BCR’s precise role in germinal center selection has been difficult to assess: mechanistic investigations into its modulation during the decision of B cell fate suffer from experimental imitations due to the fundamental role of the BCR in B cell biology. Bruton’s tyrosine kinase (BTK), a member of the TEC family, is a key component of BCR signaling. Using a computational screening method based on small molecule docking outside of the BTK active site, putative allosteric modulators of BTK have been identified. In vitro primary B cell Nur777 upregulation, calcium flux, proliferation and plasma cell differentiation confirmed that these molecular probes alter B cell activation strength, kinetics, and fate only when the BCR is engaged. In vivo immunization using an antigen model revealed that BTK agonism strongly favors B cell differentiation into switched plasma cells. BTK agonism, either at the time of the immunization or at the peak of germinal center formation, enhanced humoral immunity, providing for the first time that transient, augmented BCR signaling during germinal center reaction has a direct impact on B cell differentiation ability. In addition to being powerful tools for basic B cell biology investigation, allosteric agonists of BTK could be developed into antigen-licensed B cell adjuvants for vaccines in at risk populations, as well as into cancer immunotherapy drugs involving enhancement of hyperactive B cells in tumor microenvironments during checkpoint inhibitor blockade therapies.

[0214] Computation screen allowed for the identification of several BTK modulators which worked to either inhibit or enhance BCR signaling in vitro.

[0215] Treatment with modulator sustained BCR signaling with the presence of BTK active site inhibitor indicated the compound’s dominant ability to maintain a more open confirmation for BTK.

[0216] Enhancing BCR signaling through BTK modulation led to more efficient plasma cell output in vitro and allowed for bolstered germinal center reactions which resulted in a larger proportion of switch plasma cells in vivo. Example 2: TEC kinase agonists and their use in enhancing lymphocyte activation Background

[0217] This disclosure provides an overview of the discovery of 12 small molecule allosteric agonists of the non-receptor TEC Tyrosine kinase family of lymphocyte signal transducers. Effort was focused on discovering specific modulators of Bruton’s Tyrosine Kinase (BTK) and IL-2 inducible T cell Kinase (ITK), predominantly expressed in B and T lymphocytes, respectively. BTK is a validated drug target, as exemplified by Ibrutinib, a high profile BTK inhibitor used to treat B cell tumors and GVHD. Ibrutinib is a covalent binder to the BTK active site and interferes with binding of the fuel (ATP) to the enzyme. This mode of inhibition applies to nearly all kinase inhibitors in the clinics. ITK is a known integrator of signals from T cell antigen receptor (TCR) and the costimulatory molecule CD28, the two pivotal T cell surface proteins absolutely required to activate naive T cells. ITK has been considered as a possible signal switch involved in promoting asthma, leading to the development of ITK inhibitors to treat the disease. This initial effort was clinically ineffective. Scientific rationale for the selection of asthma as the disease in which ITK plays a pivotal role has eroded with studies that have pointed to a broadly critical function of ITK in T cell activation. Recent focus has switched to testing ITK inhibition in specific T cell tumor types and early clinical trials are on-going.

[0218] All current efforts in the ITK and BTK modulation field have been to generate effective inhibitors. The same focus exists for all intracellular kinases implicated in diseases. The present disclosure describes an entirely new class of small molecule drugs to activate TEC kinases in the immune system.

[0219] Description

[0220] Discussed herein is a structure-based drug design and high-throughput in silico and cellular screening workflow to identify novel allosteric BTK and ITK agonists. Initial hits that showed robust B and / or T cell activating signals in ex vivo lymphocyte assays underwent an initial phase of medicinal chemistry to generate more potent variants. Identified herein are molecules that ramp up B and T cell activation in primary ex vivo lymphocytes and in an animal model of T cell-dependent, B cell activation and antibody production against foreign proteins (similar to a vaccination model).

[0221] These molecules are for treatment of multiple tumor types, including the breadth of solid cancers currently being treated by the class of “Immune Checkpoint Blockade” (ICB) drugs. The paradigm shift in the treatment landscape of cancer in the last two decades has been the acceptance of T cells (and B cells) as the primary defenders against cancer cells. Cancer is therefore a consequence of breakdowns in the constant immuno surveillance of cell transformation. Overwhelming exposure to cancerous cells can render T cells hyporesponsive and tip the balance to cancer progression. Blocking cell surface molecules (e.g., CTLA-4, PD-1) on T cells that normally dampen CD28 signaling unleash T cells to kill tumors. The inhibitory receptors such as CTLA-4 and PD-1 are termed checkpoint inhibitors (CI) and the immunotherapy against them are referred to ICB. Treatment of cancers has been transformed by ICB starting in 2011. More than 20% of cancer therapies now are ICB drugs and by 2030 they are on pace to grow to >40%. ICB therapeutics train the patients’ immune system to control cancer and for several indications cancer “cure” has been achieved. Despite this remarkable progress the sobering reality is that for most immunogenic solid tumors, the efficacy of ICB drugs are in low double-digit percentages and most patients do not durably benefit. Moreover, ICB drugs can cause severe, but clinically manageable, side-effects leading to substantial rates of discontinuation. To overcome these central hurdles in efficacy and safety, there have been intensive efforts by the biomedical community to identify synergistic modalities to enhance ICB efficacy and reduce toxicity. While steady progress has been made in specific tumor settings, these efforts have not translated into a breakthrough status across ICB-sensitive tumor types. The allosteric TEC kinase agonists have the potential to end the current stalemate in ICB therapy. These novel small molecules have utility in treating ICB-sensitive (immunogenic) solid tumors. These indications include, but are not limited to, lung, prostate, colon, breast, kidney, and head & neck cancers.

[0222] Current leading ICB therapies block T cell surface molecules that dampen CD28 signaling critical for activating T cells. Normal CD28 signaling and function requires ITK. The ITK agonists herein directly enhance CD28 signals and represent a specific method to enhance CD28 signaling inside T cells to generate the most fit cells for killing tumors. Cellular, molecular and biochemical assays of agonist- treated T and B cells support the contention that the agonist activated lymphocytes are endowed with molecular properties of cancer-killing lymphocytes found in patients treated with ICB drugs. In sum, a combination of a ICB drug that indirectly turns on CD28 by removing restrainers, and the agonists disclosed herein that directly enhances CD28 signaling from the inside, increased the proportion of ICB responders across cancer types and responders should achieve a longer duration of benefit. As to the issue of safety in immunotherapeutic settings, ITK is a T cell specific kinase, making it an ideal target for therapeutics aimed at changing T cell function.

[0223] First, these allosteric compounds work differently than the current ITK and BTK modulators in clinics or clinical trials that target the catalytic site located in the kinase domain. Limitations observed in the clinical setting for the covalent active site modulators include development of resistance mutations in the cancers that render the therapeutics ineffective and leads to disease relapse. The molecules described in this disclosure did not target the catalytic site which is highly conserved across multiple families of kinases. Rather, they targeted previously unexplored accessible protein surfaces of TEC kinases.

[0224] Second, compared to other potential immuno-therapeutics targeting the tumor microenvironment (TME), this target, ITK, is specific to T cells. While ITK is also expressed in NK cells, another TEC kinase RLK dominates in function in mouse NK cells. ITK transcripts are detectable by PCR in mast cells, but ITK protein expression is not consistently observed. ITK is also expressed in innate lymphoid cells Type 2 (ILC2). ITK promotes survival of intestinal ILC2, which in turn ensure gut health in mice. Hence, agonism of ITK promoted activation of T cells, but did not cause other cells of the immune system to cause damage. Unlike monoclonal antibody biologies targeting cell surface receptors that dominate ICB therapeutics, these compounds are designed to dial up an important signal transducer of CD28 inside the cells only when T cells see tumor specific moieties (antigens) via their T cell antigen receptor (TCR— the key that turns on T cells). These allosteric compounds do not covalently modify TEC kinases and predicted fast off- rates in target interaction will limit sustained changes in T cell signaling, more closely mimicking the natural high-affinity T cell to tumor target interactions. These features support the thesis that direct ITK agonism to enhance TCR-CD28 signaling in tumorspecific T cells will avoid life-degrading side-effects of the ICB drugs.

[0225] Example 1A: B cell and T cell Activity

[0226] B cell Ca+2flux assay

[0227] For B cell calcium flux splenic B cells were isolated using the EasySep B cell isolation kit and treated similar to the T cells. At the flow cytometer cells were stimulated with lOpg / mL anti-IgM F(ab’)2 while data acquisition for 5 minutes. Data was processed and quantified in FlowJo. (Table 1) B cell Nur77 expression assay

[0228] Splenic B cells were isolated from Nur77 reporter mice by negative selection (EasySep kit) and stimulated with Ipg / mL anti-IgM antibody for 18 hours in the presence of DMSO (untreated) or compounds at indicated concentrations. Harvested cells were resuspended in FACS buffer and run on a BD SORP. Data was analyzed on FlowJo. (Table 1)

[0229] The compounds in Tables 1 and 2 were prepared using multi-step synthesis pathways, such as those disclosed in Figures 7-9, and available chemical precursors.

[0230] Table 1. B cell activity

[0231]

[0232] Surface Marker Expression

[0233] Surface marker expression testing or activity was tested via flow cytometric analyses of T cell activation markers including, but not limited to, CD3e, CD4, CD8, TCR|3, CD25, CD44, CD62L, CD69, CD101, CD226, PD-1, Slamf6, Tim-3, CXCR3 (Table 2). T cell Ca2+flux assay

[0234] CD4+ T cells were enriched from C57 / B6 mouse splenocytes according to manufacturer’s protocol (EasySepCD4 T cell isolation kit). Total cells of interest were incubated in 4uM Fluo-3 (ThermoFisher) and lOuM FuraRed (ThermoFisher) in HBSS (Coming) with 10% Pluronic Acid F-68 (Gibco) for 30 minutes in the dark at 37°C followed by incubation with compounds of interest in HBSS +2% FCS buffer for 45 minutes in the dark at 37°C. Cells were stained in HBSS +2% FCS using Fixable Viability Dye, CD4 (RM4-5), and CD8 alpha (53-6.7) for 10 minutes at room temperature in the dark. For flow cytometry calcium flux was stimulated by the addition of lOug / mL CD3 biotin (145-2C11) to samples being actively recorded followed by lOug / mL purified Streptavidin. Data was processed and quantified in FlowJo (Table 2).

[0235] For B cell calcium flux splenic B cells were isolated using the EasySep B cell isolation kit and treated similar to the T cells. At the flow cytometer cells were stimulated with lOpg / mL anti-IgM F(ab’)2 while data acquisition for 5 minutes. Data was processed and quantified in FlowJo (Table 2).

[0236] Table 2. T cell activity.

[0237] Other advantages which are obvious, and which are inherent to the invention, will be evident to one skilled in the art. It will be understood that certain features and subcombinations are of utility and may be employed without reference to other features and sub-combinations. This is contemplated by and is within the scope of the claims. Since many possible embodiments may be made of the invention without departing from the scope thereof, it is to be understood that all matter herein set forth or shown in the accompanying drawings is to be interpreted as illustrative and not in a limiting sense.

Claims

CLAIMSWhat is claimed is:

1. A compound according to Formula I:(Formula I) or a pharmaceutically acceptable salt thereof; whereinX is O, S, or N(H);R1is H or Ci-20 alkyl;R2is a halogen; each RAis independently OH, halogen, C1-20 alkyl, C1-20 alkoxy, amino, C1-20 alkylamino, C2-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, or 7-20 membered alkylaryl; n is 0, 1, 2, or 3;R3is H, Ci -20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-20 acyl, C1-20 alkoxy, 7-20 membered aryloxy, C1-20 alkylamino, C2-20 dialkylamino, halogen, or amino; and if X is S and R3is -OCHF2, then R2is H, Br, or F.

2. The compound of claim 1, wherein R1is CH3 and X is S.

3. The compound of claim 1, wherein R1is CH3 and X is O.

4. The compound of claim of any one of claims 1-3, wherein R3is-OCHF2.

5. The compound of any one of claims 1-4, wherein R2is Cl.

6. The compound of any one of claims 1-5, wherein n is 0.

7. The compound of any one of claims 1-6, wherein at least one RAis methyl or both RAare methyl and n is 2.

8. The compound of any one of claims 6-7, wherein R3is -CO2H and R1is not methyl.

9. The compound of any one of claims 6-7, wherein R3is OH and RAis not -OCH3.

10. A compound according to Formula II:or a pharmaceutically acceptable salt thereof; whereinZ is O, S, or N(H);R6is a halogen;R4is substituted or unsubstituted C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-20 acyl, C1-20 alkoxy, 7-20 membered aryloxy, C1-20 alkylamino, C2-20 dialkylamino, halogen, or amino; and11. The compound of claim 10, wherein R6is Cl.

12. The compound of any one of claims 10-11, wherein R4is -CHF2.

13. The compound of any one of claims 10-12, R4is -CHF2 and R5is14. The compound of any one of claims 10-11, wherein R4is -C(O)CH3.

15. A compound according to Formula III:(Formula III) or a pharmaceutically acceptable salt thereof; whereinA is O, S, or N(H);R8is H or halogen;R9is C1-20 alkyl, 3-20 membered cycloalkyl, or 6-20 membered aryl;with the proviso that (a) when16. The compound of claim 15, wherein A is S and R9is pentyl.

17. The compound of claim 15, wherein A is O and R9is methyl.

18. The compound of claim 16, wherein19. The compound of claim 17, wherein20. A compound according to Formula IV :or a pharmaceutically acceptable salt thereof; whereinwith the proviso that when, then R10isThe compound of claim 20, wherein n is 0 or 1.

22. The compound of claim 20 or 21, wherein23. The compound of claim 20 or 21, wherein M is S and R11is phenyl.

24. The compound of claim 20 or 21, wherein M is O and R11is phenyl.

25. The compound of claim 20 or 21, wherein M is S and R10is28. The compound of claim 27, wherein R11is phenyl.

29. The compound of claim 27 or 28, wherein n is 1.

30. A method of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula V:or a pharmaceutically acceptable salt thereof, whereinX is O, S, or N(H);R1is H or substituted or unsubstituted C1-20 alkyl;R2is H or halogen;Each RAis independently OH, halogen, C1-20 alkyl, C1-20 alkoxy, amino, C1-20 alkylamino, C2-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, or 7-20 membered alkylaryl; n is 0, 1, 2, or 3;R3is H, substituted or unsubstituted C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-20 acyl, C1-20 alkoxy, 7-20 membered aryloxy, C1-20 alkylamino, C2-20 dialkylamino, halogen, or amino.

31. The method of claim 30, wherein R1is CH3 and X is S.

32. The method of claim 30, wherein R1is CH3 and X is O.

33. The method of any one of claims 30-32, wherein R3is -OCHF2.

34. The method of any one of claims 30-33, wherein R2is Cl.

35. The method of any one of claims 30-34, wherein n is 0.

36. The method of any one of claims 30-35, wherein at least one RAis methyl or both RAare methyl and n is 2.

37. The method of any one of claims 35-36, wherein R3is -CO2H and R1is not methyl.

38. The method of any one of claims 35-36, wherein R3is OH and RAis not -OCH3.

39. A method of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula VI:(Formula VI), or a pharmaceutically acceptable salt thereof; whereinZ is O, S, or N(H);R6is a halogen;R4is substituted or unsubstituted C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-20 acyl, C1-20 alkoxy, 7-20 membered aryloxy, C1-20 alkylamino, C2-20 dialkylamino, halogen, or amino; and40. The method of claim 39, wherein R6is Cl.

41. The method of any one of claims 39-40, wherein R4is CHF2.

42. The method of any one of claims 39-41, wherein R4is CHF2 and R5is43. The method of any one of claims 39-41, wherein R4is CHF2 and R5is44. The method of any one of claims 39-40, wherein R4is -C(O)CH3.

45. A method of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula VII:(Formula VII), or a pharmaceutically acceptable salt thereof; whereinA is O, S, or N(H);R8is H or halogen;R9is Ci-20 alkyl, 3-20 membered cycloalkyl, or 6-20 membered aryl; andwith the proviso that whenis halogen.

46. The method of claim 45, wherein A is S and R9is pentyl.

47. The method of claim 45, wherein A is O and R9is methyl.

49. The method of claim 47, wherein50. A method of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula VIII:(Formula VIII), or a pharmaceutically acceptable salt thereof; wherein51. The method of claim 50, wherein n is 0 or 1.

52. The method of claim 50 or 51, wherein53. The method of claim 50 or 51, wherein M is S and R11is phenyl.

54. The method of claim 50 or 51, wherein M is O and R11is phenyl.

55. The method of claim 50 or 51, wherein M is S and R10is58. The method of claim 57, wherein R11is phenyl.

59. The method of claim 57 or 58, wherein n is 1.

60. The method of any one of claims 30-59, wherein cancer comprises lung cancer, prostate cancer, colon cancer, breast cancer, kidney cancer, or head and neck cancer.

61. The method of any one of claims 30-60, wherein the cancer comprises Stage I, Stage II, Stage III, or Stage IV cancer.

62. The method of any one of claims 30-61, further comprising administering a targeted therapy.

63. The method of claim 62, wherein the targeted therapy comprises a small molecule drug, a monoclonal antibody, or any combination thereof.

64. The method of any one of claims 30-63, wherein the method further comprises administering chemotherapy, radiation therapy, stem cell transplantation, or any combination thereof.

65. A method of reducing solid tumor size comprising administering a therapeutically effective amount of a compound according to any one of claims 31-60, or a pharmaceutically acceptable salt thereof.

66. The method of claim 65, wherein the solid tumor comprises cancer cells, cancer stem cells, connective-tissue cells, immune cells, or any combination thereof.

67. The method of claim 65 or 66, wherein the solid tumor is a carcinoma, a sarcoma, a lymphoma, a blastoma, a melanoma, a germ cell tumor, or a carcinosarcoma.

68. The method of any one of claims 65-67, wherein the solid tumor is a result of breast cancer, lung cancer, prostate cancer, colon cancer, or head and neck cancer.

69. The method of any one of claims 65-68, wherein the method further comprises administering chemotherapy, radiation therapy, stem cell transplantation, or any combination thereof.

70. A method of increasing lymphocyte activation comprising subjecting the lymphocyte to a compound of any one of claims 30-59.

71. A method of activating Tec kinase comprising targeting a surface of the Tec kinase with any one of the compounds of claims 30-59.

72. The method of claim 71, wherein the Tec kinase comprises Bruton’s tyrosine kinase (BTK).

73. The method of claim 71, wherein the Tec kinase comprises IL-2 inducible Tcell kinase (ITK).

74. A method of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula IA, or a pharmaceutically acceptable salt thereof:(Formula IA), whereinX is S or O;R1is 6-20 membered aryl, 7-20 membered alkylaryl, or 3-20 membered cycloalkyl;R2is H, substituted or unsubstituted C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-C20 acyl, C1-C20 alkoxy, or 7-20 membered aryloxy; each RAis independently OH, halogen, C1-20 alkyl, C1-20 alkoxy, amino, C1-20 alkylamino, C2-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, or 7-20 membered arylalkyl, or two RAgroups adjacent to one another together with atoms to which they are attached to form a fused 5 to 7 membered cyclic moiety; and n is 0, 1, or 2.

75. The method of claim 74, wherein R1is a 3 to 7 membered aryl optionally substituted with from 1 to 3 RAgroups.

77. The method of any one of claims 74-76, wherein R2is a Ci-4 alkyl optionallysubstituted with from 1 to 3 RAgroups.

78. The method of claim 72, wherein R2is methyl, difluoromethyl, trifluoromethyl, or fluoromethyl.

79. The method of any one of claims 74-76, wherein R2is a cycloalkyl optionally substituted with from 1 to 3 RAgroups.

80. A method of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula IIA, or a pharmaceutically acceptable salt thereof:whereinR3is H or acyl;R4and R5are independently of each other H, C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl,6-20 membered aryl, 7-20 membered alkylaryl, or 3-20 membered cycloalkyl, or R4and R5together with the nitrogen atom to which they are attached, form a 5 to 10 membered cyclic moiety; each RAis independently C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl,7-20 membered alkylaryl, or 3-20 membered cycloalkyl; and n is 0, 1, or 2.

81. The method of claim 80, wherein R4and R5are adjacent to one another and together with the nitrogen carbon atom to with they are attached form a 9 membered cyclic moiety.

82. The method of claim 80 or 81, wherein at least one of R4and R5comprisesRA.

83. The method of claim 82, wherein the 9 membered cyclic moiety and RAare adjacent to one another and together with atoms to which they are attached form a fused 13 membered cyclic moiety.

84. The method of any one of claims 80-83, wherein at least one of R4and R5comprise:

85. The method of any one of claims 80-84, wherein R3comprises an alkylacyl or a cycloalkyl acyl.

86. A method of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula IIIA, or a pharmaceutically acceptable salt thereof:whereinX is S or O; each RAis independently OH, halogen, C1-20 alkyl, Ci-2oalkoxy, amino, C1-20 alkylamino, C1-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, or 7-20 membered arylalkyl, or two RAgroups adjacent to one another together with atoms to which they are attached to form a fused 5 to 10 membered cyclic moiety; and n is 0, 1, or 2.

87. The method of claim 86, wherein RAcomprises:

88. A method of treating cancer in a subject comprising administering a therapeutically effective amount of a compound according to Formula IVA, or a pharmaceutically acceptable salt thereof:whereinR6is H, OH, halogen, Ci -20 alkyl, C2-20 alkenyl, C2-20 alkynyl, 6-20 membered aryl, 7-20 membered alkylaryl, 3-20 membered cycloalkyl, C1-C20 acyl, C1-C20 alkoxy, or 7-20 membered aryloxy; each RAis independently OH, halogen, Ci-2oalkyl, Ci-2oalkoxy, amino, C1-20 alkylamino, C2-20 dialkylamino, 3-20 membered cycloalkyl, 6-20 membered aryl, or 7-20 membered arylalkyl, or two RAgroups adjacent to one another together with atoms to which they are attached to form a fused 5 to 10 membered cyclic moiety; and n is 0, 1, or 2.

89. The method of claim 88, wherein R6further comprises RA, further wherein R6is a cycloalkyl, aryl, or alkylaryl and is adjacent to RAand together with the atoms to which R6and RAare attached form a fused 5 to 10 membered cyclic moiety.

90. The method of claim 88 or 89, wherein R6comprises:

91. The method of claim 88, wherein RAis Ci-6 alkyl.

92. The method of claim 91, wherein n is 1 and RAis methyl.

93. The method of any one of claims 74-92, wherein cancer comprises lung cancer, prostate cancer, colon cancer, breast cancer, kidney cancer, or head and neck cancer.

94. The method of any one of claims 74-93, wherein the cancer comprises Stage I, Stage II, Stage III, or Stage IV cancer.

95. The method of any one of claims 74-94, further comprising administering a targeted therapy.

96. The method of claim 95, wherein the targeted therapy comprises a small molecule drug, a monoclonal antibody, or any combination thereof.

97. The method of any one of claims 74-96, wherein the method further comprises administering chemotherapy, radiation therapy, stem cell transplantation, or any combination thereof.

98. A method of reducing solid tumor size comprising subjecting the tumor to a compound disclosed in any one of claims 74-92.

99. The method of claim 98, wherein the solid tumor comprises cancer cells, cancer stem cells, connective-tissue cells, immune cells, or any combination thereof.

100. The method of claim 98 or 99, wherein the solid tumor is a carcinoma, a sarcoma, a lymphoma, a blastoma, a melanoma, a germ cell tumor, or a carcinosarcoma.

101. The method of any one of claims 98-100, wherein the solid tumor is a result of breast cancer, lung cancer, prostate cancer, colon cancer, or head and neck cancer.

102. The method of any one of claims 98-101, wherein the method further comprises administering chemotherapy, radiation therapy, stem cell transplantation, or any combination thereof.

103. A method of increasing lymphocyte activation comprising subjecting the lymphocyte cell to a compound disclosed in any one of claims 74-92.

104. A method of activating TEC kinase comprising targeting a TEC kinase with a compound disclosed in any one of claims 74-92.

105. The method of claim 104, wherein the TEC kinase comprises Bruton’s tyrosine kinase (BTK).

106. The method of claim 104, wherein the TEC kinase comprises IL-2 inducible T cell kinase (ITK).

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