Use of ITK inhibitors and car t-cell therapy

Abstract

Interleukin-2-inducible T-cell kinase inhibitors are useful and efficacious in methods of treating cancer, methods for preventing and / or reversing CAR T-cell exhaustion, methods for stimulating CAR T-cell-mediated immune responses, methods for increasing CAR T-cell function, and method for increasing expression of T cell factor- 1 on CAR T-cells. An exemplary interleukin- 2-inducible T-cell kinase inhibitor is soquelitinib.

Description

PATENT Attorney Docket No.048517-558001WO USE OF ITK INHIBITORS AND CAR T-CELL THERAPY CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to US Application No.63 / 611,539 filed December 18, 2023, the disclosure of which is incorporated by reference herein in its entirety. BACKGROUND

[0002] T cells are immune cells that become activated via T cell receptor (TCR) signaling and co-stimulation following engagement with antigen. Physiologic activation through the T cell receptor renders T cells capable of mediating potent antitumor and / or anti-infective effects. During resolution of an acute inflammatory response, a subset of activated effector T cells differentiate into long-lived memory cells. By contrast, in patients with chronic infections or cancer, T cells may undergo pathologic differentiation toward a state of dysfunction, which has been termed T cell exhaustion. T cell exhaustion is characterized by marked changes in metabolic function, transcriptional programming, loss of effector function (e.g., cytokine secretion, killing capacity), and co-expression of multiple surface inhibitory receptors. The root cause of T cell exhaustion is persistent antigen exposure leading to continuous TCR signaling. Prevention or reversal of T cell exhaustion has been long sought as a means to enhance T cell effectiveness (e.g., in patients with cancer). The disclosure is directed to this, as well as other, important ends. BRIEF SUMMARY

[0003] Provided herein are methods of treating cancer, preventing and / or reversing CAR T-cell exhaustion, stimulating a CAR T-cell-mediated immune response, increasing CAR T-cell function, and increasing expression of T cell factor-1 (Tcf1) on CAR T-cells in a patient in need thereof comprising administering to the patient: (i) an effective amount of an interleukin-2- inducible T-cell kinase (ITK) inhibitor and (ii) an effective amount of CAR T-cells.

[0004] Provided herein are methods of treating cancer, preventing and / or reversing CAR T-cell exhaustion, stimulating a CAR T-cell-mediated immune response, increasing CAR T-cell function, and increasing expression of Tcf1 on CAR T-cells in a patient in need thereof comprising: (i) contacting CAR T-cells with an effective amount of an ITK inhibitor in vitro, thereby producing ITK-contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK-contacted CAR T-cells. In embodiments, the methods further comprise administering to the patient an effective amount of the ITK inhibitor.

[0005] Provided herein are methods of enhancing the properties of CAR T-cells comprisingcontacting CAR T-cells with an ITK inhibitor, thereby producing an ITK-contacted CAR T-cell. Enhance properties of ITK-contacted CAR T-cells include increased expression of Tcf1.

[0006] Provided herein are compositions and kits comprising ITK inhibitors and CAR T-cells.

[0007] These and other embodiments and aspects are described in detail herein. BRIEF DESCRIPTION OF THE DRAWING

[0008] FIGS.1A-1G show that soquelitinib inhibits T cell exhaustion process in vitro. FIG. 1A: Schematic displaying the experimental design for evaluating the effect of soquelitinib on OT-1 T cell exhaustion process. Soquelitinib inhibits the expression of several markers associated with T cell exhaustion in a dose-dependent manner. FIG.1B: Representative histogram overlays of indicated exhaustion marker expression in CD8+Va2+ T cells are shown for day 7. FIG.1C: Summary graph of two exhaustion markers, lymphocyte activation protein (LAG)3 and T cell immunoreceptor with Ig and ITIM domains (TIGIT) is shown. FIG.1D: Schematic describing the experimental design for evaluating the effect of soquelitinib in restoring T cell exhaustion in vitro. Soquelitinib dose-dependently (up to 1 mM) restores the potential functions of OT-1 CD8 T cells as indicated by increased production of Granzyme B and IFNg production following the 4d treatment period. FIG.1E: Representative histograms of Granzyme B and IFNg from OT-1 T cells treated with indicated concentration of soquelitinib or DMSO control are shown. FIGS.1F-1G: Summary graph of Granzyme B and IFNg-producing OT-1 T cells is shown. OVA=ovalbumin; APC=antigen presenting cell; MFI=mean fluorescence intensity. SQL=soquelitinib.

[0009] FIG.2 shows that soquelitinib (SQL) reduces exhaustion and increases expression of T-cell factor-1 (TCF1) on CD8 T cells (tumor infiltrating lymphocytes) in mouse tumor EL4 tumors and in uninvolved spleen. EL4 tumor-bearing mice were treated with either vehicle or solution-formulated soquelitinib (30 mg / kg, or 10 mg / kg) for 8 days. PD1highTCF1+ CD8 TILs were assessed by flow cytometry. TCF1 is a transcription factor that increases self renewal capacity of CD8 T cells and increases formation of memory CD8 cells. TCF1 on CART has been reported to be one of the most important factors for CART success in patients.

[0010] FIG.3 shows the selective binding of ibrutinib and soquelitinib to various kinase receptors. DETAILED DESCRIPTION

[0011] Definitions

[0012] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art. See, e.g., Singleton etal., Dictionary of Microbiology and Molecular Biology 2nd ed., J. Wiley & Sons (New York, NY 1994); Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Springs Harbor Press (Cold Springs Harbor, NY 1989). Any methods, devices and materials similar or equivalent to those described herein can be used. The following definitions are provided to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present disclosure.

[0013] The terms “interleukin-2-inducible T-cell kinase” and “ITK” refer to a protein (including homologs, isoforms, and functional fragments thereof) with interleukin-2-inducible T-cell kinase activity. The term includes any recombinant or naturally-occurring form of ITK or variants thereof that maintain ITK activity (e.g. within at least 60%, 70%, 80%, 90%, or 100% activity compared to wildtype ITK). In aspects, the interleukin-2-inducible T-cell kinase protein encoded by the ITK gene has the amino acid sequence set forth in or corresponding to Entrez 3702, UniProt Q08881, or RefSeq (protein) NP_005537. In aspects, the ITK gene has the nucleic acid sequence set forth in RefSeq (mRNA) NM_005546. In aspects, the sequence corresponds to GI: 15718680, to NP_005537.3, to NM_005546.3, or to GI: 21614549.

[0014] The term “inhibition,” “inhibit,” “inhibiting” and the like in reference to a protein- inhibitor interaction means negatively affecting (e.g. decreasing) the activity or function of the protein relative to the activity or function of the protein in the absence of the inhibitor. In embodiments, inhibition means negatively affecting (e.g. decreasing) the concentration or levels of the protein relative to the concentration or level of the protein in the absence of the inhibitor. In embodiments, inhibition refers to reduction of a disease or symptoms of disease. In embodiments, inhibition refers to a reduction in the activity of a particular protein target. Thus, inhibition includes, at least in part, partially or totally blocking stimulation, decreasing, preventing, or delaying activation, or inactivating, desensitizing, or down-regulating signal transduction or enzymatic activity or the amount of a protein. In embodiments, inhibition refers to a reduction of activity of a target protein resulting from a direct interaction (e.g. an inhibitor binds to the target protein). In embodiments, inhibition refers to a reduction of activity of a target protein from an indirect interaction (e.g. an inhibitor binds to a protein that activates the target protein, thereby preventing target protein activation).

[0015] An “interleukin-2-inducible T-cell kinase inhibitor” and “ITK inhibitor” is a compound that negatively affects (e.g. decreases) the activity or function of interleukin-2-inducible T-cell kinase relative to the activity or function of interleukin-2-inducible T-cell kinase in the absence of the inhibitor (e.g., wherein the ITK inhibitor binds ITK).

[0161] “Soquelitinib” refers to an ITK inhibitor having the following chemical structure or apharmaceutically acceptable salt thereof: O O .

[0016] “T-cellmay occur as a result of an infection (e.g., a chronic infection) or a disease (e.g., cancer). T-cell exhaustion is associated with increased expression of PD-1, TIM-3, and LAG-3, apoptosis, and reduced cytokine secretion. Accordingly, the terms “reverse T-cell exhaustion” and “reduce T-cell exhaustion” refer to a condition of restored functionality of T-cells characterized by one or more of the following: decreased expression and / or level of one or more of PD-1, TIM-3, and LAG-3; increased memory cell formation and / or maintenance of memory markers (e.g., CD62L); prevention of apoptosis; increased antigen-induced cytokine (e.g., IL-2) production and / or secretion; enhanced cytotoxicity / killing capacity; increased recognition of tumor targets with low surface antigen; enhanced proliferation in response to antigen.

[0017] The term “autologous” is meant to refer to any material (e.g., T-cells) derived from the same individual to which it is later to be re-introduced into the individual. The term “allogeneic” refers to a biological material derived from a different animal of the same species. “Xenogeneic” refers to a graft derived from an animal of a different species.

[0018] A “CAR-T cell” is a T cell that expresses a chimeric antigen receptor. In embodiments, The T-cell is a CD4+ T-cell, a CD8+ T-cell, or a combination thereof. The phrase “chimeric antigen receptor” or “CAR” as generally used in the art refers to a recombinant fusion protein that has an antigen-specific extracellular domain coupled to an intracellular domain that directs the cell to perform a specialized function upon binding of an antigen to the extracellular domain. Chimeric antigen receptors are distinguished from other antigen binding agents by their ability to both bind MHC-independent antigen and transduce activation signals via their intracellular domain. The antigen-specific extracellular domain of a chimeric antigen receptor recognizes and specifically binds an antigen, typically a surface-expressed antigen of a malignancy. An antigen- specific extracellular domain specifically binds an antigen when, for example, it binds the antigen with an affinity constant or affinity of interaction (KD) between about 0.1 pM to about10 μΜ. An antigen-specific extracellular domain suitable for use in a CAR may be any antigen- binding polypeptide, a wide variety of which are known in the art. In some instances, the antigen-binding domain is a single chain Fv (scFv). Other antibody based recognition domains (cAb VHH (camelid antibody variable domains) and humanized versions thereof, IgNAR VH (shark antibody variable domains) and humanized versions thereof, sdAb VH (single domain antibody variable domains) and “camelized” antibody variable domains are suitable for use. In some instances, T-cell receptor (TCR) based recognition domains such as single chain TCR (scTv, single chain two-domain TCR containing VαVβ) are also suitable for use. Suitable antigens may include T cell-specific antigens and / or antigens that are not specific to T cells. In a preferred embodiment, an antigen specifically bound by the chimeric antigen receptor of a CAR- T cell, and the antigen for which the CAR-T cell is deficient, is an antigen expressed on a malignant T cell, more preferably an antigen that is overexpressed on malignant T cell in comparison to a non- malignant T cell.

[0019] The term “antigen” or “cancer antigen” refers to peptides, proteins, or fragments thereof expressed on the surface of a cancer cell that are capable of binding to the antibody binding domain provided herein. In embodiments, a antigen binding domain binds to a tumor- associated antigen. In embodiments, a antigen binding domain binds to a tumor-specific antigen. In embodiments, the antigen binding domain binds to a surface protein expressed by cells present within a malignant tumor. The vaccine can be prepared from the causative agent of a disease (in this case, the tumor itself or the cells contained within the tumor), its products, or a synthetic substitute, treated to act as an antigen without inducing the disease. The antigens are substances on the surface of cells that are not normally part of the body. The immune system is stimulated by the vaccine to attack the antigens, usually getting rid of them. This leaves the immune system with a “memory” that helps it respond to those antigens in the future. Cancer treatment vaccines boost the immune system's ability to recognize and destroy antigens present on the cancer cells. Cancer cells often have certain molecules called cancer-specific antigens on their surface that healthy cells do not have. When these molecules used to manufacture a vaccine, the molecules act as antigens. The vaccine then stimulates the immune system to recognize and destroy cancer cells that have these molecules on their surface. Many cancer vaccines also contain adjuvants, which are substances that may help strengthen the immune response. In this embodiment, the cancer vaccines are manufactured to target the surface antigens present individual patient’s tumor. This type of vaccine is produced from the cells acquired form the person's tumor sample, and then stress treated to ultimately generate large populations of cancer stem cells and all of their progeny including the more mature cancer cells.This enables the manufacture of an effective vaccine from a small biopsy of the tumor, rather than necessitating surgery to get a large enough sample of the tumor to create the vaccine, as is the practice with other cancer vaccines.

[0020] The term “protein” refers to a polymer of amino acid residues, wherein the polymer may be conjugated to a moiety that does not consist of amino acids. The term applies to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers.

[0021] The term “pharmaceutically acceptable salts” is meant to include salts of active compounds that are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein. When compounds disclosed herein contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. When compounds disclosed herein contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, oxalic, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, for example, Berge et al., “Pharmaceutical Salts”, Journal of Pharmaceutical Science, 66:1-19 (1977)).

[0022] An “effective amount” is an amount sufficient to accomplish a stated purpose (e.g., achieve the effect for which it is administered, treat a disease, reduce enzyme activity, increase enzyme activity, reduce one or more symptoms of a disease or condition). An example of an “effective amount” is an amount sufficient to contribute to the treatment, prevention, or reduction of a symptom or symptoms of a disease, which could also be referred to as a “therapeutically effective amount.” A “reduction” of a symptom or symptoms (and grammatical equivalents of this phrase) means decreasing of the severity or frequency of the symptom(s), orelimination of the symptom(s). A “prophylactically effective amount” of a drug is an amount of a drug that, when administered to a subject, will have the intended prophylactic effect, e.g., preventing or delaying the onset (or reoccurrence) of an injury, disease, pathology or condition, or reducing the likelihood of the onset (or reoccurrence) of an injury, disease, pathology, or condition, or their symptoms. The full prophylactic effect does not necessarily occur by administration of one dose, and may occur only after administration of a series of doses. Thus, a prophylactically effective amount may be administered in one or more administrations. The exact amounts will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols.1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).

[0023] The term “administering” means oral administration, administration as a suppository, topical contact, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal or subcutaneous administration, or the implantation of a slow-release device, e.g., a mini-osmotic pump, to a subject. Administration is by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, e.g., intravenous, intramuscular, intra-arteriole, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, etc. In aspects, the term administering means vaccination.

[0024] “Patient,” “subject,” “patient in need thereof,” and “subject in need thereof” are herein used interchangeably and refer to a living organism suffering from or prone to a disease or condition that can be treated by administration of a compound or vaccine as provided herein. Non-limiting examples include humans, other mammals, bovines, rats, mice, dogs, cats, monkeys, cow, and other non-mammalian animals. In aspects, a patient is a mammal. In aspects, a patient is human.

[0025] The compounds described herein can be used in combination with one another, with other active drugs known to be useful in treating a disease (e.g., chemotherapy) or with adjunctive agents that may not be effective alone, but may contribute to the efficacy of the active agent. Thus, the compounds described herein may be co-administered with one another or with other active drugs known to be useful in treating a disease.

[0026] Descriptions of compounds of the present disclosure are limited by principles of chemical bonding known to those skilled in the art. Accordingly, where a group may besubstituted by one or more of a number of substituents, such substitutions are selected so as to comply with principles of chemical bonding and to give compounds which are not inherently unstable and / or would be known to one of ordinary skill in the art as likely to be unstable under ambient conditions, such as aqueous, neutral, and several known physiological conditions. For example, a heterocycloalkyl or heteroaryl is attached to the remainder of the molecule via a ring heteroatom in compliance with principles of chemical bonding known to those skilled in the art thereby avoiding inherently unstable compounds.

[0027] The abbreviations used herein have their conventional meaning within the chemical and biological arts. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts. Where substituent groups are specified by their conventional chemical formulae, written from left to right, they equally encompass the chemically identical substituents that would result from writing the structure from right to left, e.g., -CH2O- is equivalent to -OCH2-.

[0028] The terms “a” or “an,” as used in herein means one or more. In addition, the phrase “substituted with a[n],” as used herein, means the specified group may be substituted with one or more of any or all of the named substituents. For example, where a group, such as an alkyl or heteroaryl group, is “substituted with an unsubstituted C1-C20 alkyl, or unsubstituted 2 to 20 membered heteroalkyl,” the group may contain one or more unsubstituted C1-C20alkyls, and / or one or more unsubstituted 2 to 20 membered heteroalkyls.

[0029] The term “alkyl,” by itself or as part of another substituent, means, unless otherwise stated, a straight (i.e., unbranched) or branched carbon chain (or carbon), or combination thereof, which may be fully saturated, mono- or polyunsaturated and can include mono-, di- and multivalent radicals. The alkyl may include a designated number of carbon atoms (e.g., C1-C10 means one to ten carbons). Alkyl is an uncyclized chain. Examples of saturated hydrocarbon radicals include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n- butyl, t-butyl, isobutyl, sec-butyl homologs and isomers of, for example, n-pentyl, n-hexyl, n- heptyl, n-octyl, and the like. An unsaturated alkyl group is one having one or more double bonds or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2- propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and the higher homologs and isomers. An alkoxy is an alkyl attached to the remainder of the molecule via an oxygen linker (-O-). An alkyl moiety may be an alkynyl moiety. An alkyl moiety may be fully saturated. An alkenyl may include more than one double bond and / or one or more triple bonds in addition to the one or more double bonds. An alkynyl may include more than one triple bond and / or one or more double bonds in addition to the oneor more triple bonds.

[0030] The term “alkylene,” by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from an alkyl, as exemplified, but not limited by, -CH2CH2-. Typically, an alkyl (or alkylene) group will have from 1 to 24 carbon atoms, with those groups having 10 or fewer carbon atoms being preferred herein. A “lower alkyl” or “lower alkylene” is a shorter chain alkyl or alkylene group, generally having eight or fewer carbon atoms. The term “alkenylene,” by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from an alkene.

[0031] The term “heteroalkyl,” by itself or in combination with another term, means, unless otherwise stated, a stable straight or branched chain, or combinations thereof, including at least one carbon atom and at least one heteroatom, and wherein the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized). The heteroatom(s) may be placed at any interior position of the heteroalkyl group or at the position at which the alkyl group is attached to the remainder of the molecule. Heteroalkyl is an uncyclized chain. Examples include, but are not limited to: -CH2CH2-O-CH3, -CH2CH2-NH-CH3, -CH2-N(CH3)-CH3, -CH2-S-CH-CH3, -CH2CH2NH2-, -S(O)-CH3, -CH2-H2-S(O)2-CH3, -CH2-NH2, -CH2-NO2, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, -CH2NO2, -CH=CH-N(CH3)-CH3, -O-CH2-CH3, and -CN. Up to two or three heteroatoms may be consecutive, such as, for example, -CH2-NH-OCH3 and -CH2-O-Si(CH3)3. A heteroalkyl moiety may include one heteroatom. A heteroalkyl moiety may include two optionally different heteroatoms. A heteroalkyl moiety may include three optionally different heteroatoms. A heteroalkyl moiety may include four optionally different heteroatoms. A heteroalkyl moiety may include five optionally different heteroatoms.

[0032] Similarly, the term “heteroalkylene,” by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from heteroalkyl, as exemplified, but not limited by, -CH2-CH2-S-CH2-CH2- and -CH2-S-CH2-CH2-NH-CH2-. For heteroalkylene groups, heteroatoms can also occupy either or both of the chain termini (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, and the like). Still further, for alkylene and heteroalkylene linking groups, no orientation of the linking group is implied by the direction in which the formula of the linking group is written. For example, the formula -C(O)2R'- represents both -C(O)2R'- and -R'C(O)2-. As described above, heteroalkyl groups, as used herein, include those groups that are attached to the remainder of the molecule through a heteroatom, such as -C(O)R', -C(O)NR', -NR'R'', -OR', -SR', and / or -SO2R'. Where “heteroalkyl” is recited, followed by recitations of specific heteroalkyl groups, such as -NR'R'' or the like, it will be understoodthat the terms heteroalkyl and -NR'R'' are not redundant or mutually exclusive. Rather, the specific heteroalkyl groups are recited to add clarity. Thus, the term “heteroalkyl” should not be interpreted herein as excluding specific heteroalkyl groups, such as -NR'R'' or the like.

[0033] The terms “cycloalkyl” and “heterocycloalkyl,” by themselves or in combination with other terms, mean, unless otherwise stated, cyclic versions of “alkyl” and “heteroalkyl,” respectively. Cycloalkyl and heterocycloalkyl are not aromatic. Additionally, for heterocycloalkyl, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. In embodiments, a cycloalkyl is a spirocyclic cycloalkyl, wherein the spirocyclic rings are cycloalkyl rings. In embodiments, a cycloalkyl is a fused ring cycloalkyl, wherein the fused rings are cycloalkyl rings. In embodiments, a cycloalkyl is a bridged ring cycloalkyl, wherein the bridged rings are cycloalkyl rings. In embodiments, a bridged ring cycloalkyl refers to limitingaa cycloalkyl is three rings. In embodiments, a cycloalkyl is four rings. In embodiments, a cycloalkyl is five rings. In embodiments, a cycloalkyl is polycyclic. In embodiments, a heterocycloalkyl is a spirocyclic heterocycloalkyl, wherein the spirocyclic rings are one or more heterocycloalkyl rings and optionally one or more cycloalkyl rings. For example, spirocyclic heterocycloalkyl may refer to Inrings are one or more heterocycloalkyl rings and optionally one or more cycloalkyl rings. In embodiments,, a fused ring heterocycloalkyl a heterocycloalkyl is a bridged ringheterocycloalkyl rings and optionally one or more cycloalkyl rings. In embodiments, the rings of a spirocyclic, fused ring, or bridged ring heterocycloalkyl are heterocyclic rings. In embodiments, a heterocycloalkyl is monocyclic. In embodiments, a heterocycloalkyl is two rings. In embodiments, a heterocycloalkyl is three rings. In embodiments, a heterocycloalkyl is four rings. In embodiments, a heterocycloalkyl is five rings. In embodiments, a heterocycloalkyl is polycyclic. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples of heterocycloalkyl include, but are not limited to, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2- piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran- 3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl, and the like. A “cycloalkylene” and a “heterocycloalkylene,” alone or as part of another substituent, means a divalent radical derived from a cycloalkyl and heterocycloalkyl, respectively.

[0034] The terms “halo” or “halogen,” by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as “haloalkyl” are meant to include monohaloalkyl and polyhaloalkyl. For example, the term “halo(C1-C4)alkyl” includes, but is not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0035] The term “acyl” means, unless otherwise stated, -C(O)R where R is a substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

[0036] The term “aryl” means, unless otherwise stated, a polyunsaturated, aromatic, hydrocarbon substituent, which can be a single ring or multiple rings (preferably from 1 to 3 rings) that are fused together (i.e., a fused ring aryl) or linked covalently. A fused ring aryl refers to multiple rings fused together wherein at least one of the fused rings is an aryl ring. The term “heteroaryl” refers to aryl groups (or rings) that contain at least one heteroatom such as N, O, or S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized. Thus, the term “heteroaryl” includes fused ring heteroaryl groups (i.e., multiple rings fused together wherein at least one of the fused rings is a heteroaromatic ring). A 5,6-fused ring heteroarylene refers to two rings fused together, wherein one ring has 5 members and the other ring has 6 members, and wherein at least one ring is a heteroaryl ring. Likewise, a 6,6-fused ring heteroarylene refers to two rings fused together, wherein one ring has 6 members and the other ring has 6 members, and wherein at least one ring is a heteroaryl ring. And a 6,5- fused ring heteroarylene refers to two rings fused together, wherein one ring has 6 members andthe other ring has 5 members, and wherein at least one ring is a heteroaryl ring. A heteroaryl group can be attached to the remainder of the molecule through a carbon or heteroatom. In embodiments, an aryl is a fused ring aryl, wherein the fused rings are one or more aryl rings and optionally one or more cycloalkyl and / or heterocycloalkyl rings. In embodiments, an aryl is a bridged ring aryl, wherein the bridged rings are one or more aryl rings and optionally one or more cycloalkyl and / or heterocycloalkyl rings. In embodiments, the rings of a fused ring aryl or bridged ring aryl are aryl rings. In embodiments, an aryl is monocyclic. In embodiments, an aryl is two rings. In embodiments, an aryl is three rings. In embodiments, an aryl is four rings. In embodiments, an aryl is five rings. In embodiments, an aryl is polycyclic. In embodiments, a heteroaryl is a fused ring heteroaryl, wherein the fused rings are one or more heteroaryl rings and optionally one or more cycloalkyl, heterocycloalkyl, and / or aryl rings. In embodiments, a heteroaryl is a bridged ring heteroaryl, wherein the bridged rings are one or more heteroaryl rings and optionally one or more cycloalkyl, heterocycloalkyl, and / or aryl rings. In embodiments, the rings of a fused ring heteroaryl or bridged ring heteroaryl are heteroaryl rings. In embodiments, a heteroaryl is monocyclic. In embodiments, a heteroaryl is two rings. In embodiments, a heteroaryl is three rings. In embodiments, a heteroaryl is four rings. In embodiments, a heteroaryl is five rings. In embodiments, a heteroaryl is polycyclic. Non- limiting examples of aryl and heteroaryl groups include phenyl, naphthyl, pyrrolyl, pyrazolyl, pyridazinyl, triazinyl, pyrimidinyl, imidazolyl, pyrazinyl, purinyl, oxazolyl, isoxazolyl, thiazolyl, furyl, thienyl, pyridyl, pyrimidyl, benzothiazolyl, benzoxazoyl benzimidazolyl, benzofuran, isobenzofuranyl, indolyl, isoindolyl, benzothiophenyl, isoquinolyl, quinoxalinyl, quinolyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2- imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3- isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2- thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5- quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituents for each of the above noted aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below. An “arylene” and a “heteroarylene,” alone or as part of another substituent, mean a divalent radical derived from an aryl and heteroaryl, respectively. A heteroaryl group substituent may be -O- bonded to a ring heteroatom nitrogen.

[0037] Spirocyclic rings are two or more rings wherein adjacent rings are attached through a single atom. The individual rings within spirocyclic rings may be identical or different. Individual rings in spirocyclic rings may be substituted or unsubstituted and may have differentsubstituents from other individual rings within a set of spirocyclic rings. Possible substituents for individual rings within spirocyclic rings are the possible substituents for the same ring when not part of spirocyclic rings (e.g. substituents for cycloalkyl or heterocycloalkyl rings). Spirocylic rings may be substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heterocycloalkylene and individual rings within a spirocyclic ring group may be any of the immediately previous list, including having all rings of one type (e.g. all rings being substituted heterocycloalkylene wherein each ring may be the same or different substituted heterocycloalkylene). When referring to a spirocyclic ring system, heterocyclic spirocyclic rings means a spirocyclic rings wherein at least one ring is a heterocyclic ring and wherein each ring may be a different ring. When referring to a spirocyclic ring system, substituted spirocyclic rings means that at least one ring is substituted and each substituent may optionally be different.

[0038] The symbol “ ” and “-” denote the point of attachment of a chemical moiety to theremainder of achemical formula.

[0039] The term “oxo” means an oxygen that is double bonded to a carbon atom.

[0040] The term “alkylarylene” as an arylene moiety covalently bonded to an alkylene moiety (an alkylene linker).

[0041] An alkylarylene moiety may be substituted (e.g. with a substituent group) on the alkylene moiety or the arylene linker (e.g. at carbons 2, 3, 4, or 6) with halogen, oxo, -N3, -CF3, -CCl3, -CBr3, -CI3, -CN, -CHO, -OH, -NH2, -COOH, -CONH2, -NO2, -SH, -SO2CH3, -SO3H, -OSO3H, -SO2NH2, ^NHNH2, ^ONH2, ^NHC(O)NHNH2, substituted or unsubstituted C1-C5alkyl or substituted or unsubstituted 2 to 5 membered heteroalkyl). In embodiments, the alkylarylene is unsubstituted.

[0042] Each of the above terms (e.g., “alkyl,” “heteroalkyl,” “cycloalkyl,” “heterocycloalkyl,” “aryl,” and “heteroaryl”) includes both substituted and unsubstituted forms of the indicated radical. Preferred substituents for each type of radical are provided below.

[0043] Substituents for the alkyl and heteroalkyl radicals (including those groups often referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) can be one or more of a variety of groups selected from, but not limited to, -OR', =O, =NR', =N-OR', -NR'R'', -SR', -halogen, -SiR'R''R''', -OC(O)R', -C(O)R', -CO2R', -CONR'R'', -OC(O)NR'R'', -NR''C(O)R', -NR'-C(O)NR''R''', -NR''C(O)2R', -NR-C(NR'R''R''')=NR'''', -NR-C(NR'R'')=NR''', -S(O)R', -S(O)2R', -S(O)2NR'R'', -NRSO2R', ^NR'NR''R''', ^ONR'R'', ^NR'C(O)NR''NR'''R'''', -CN,-NO2, -NR'SO2R'', -NR'C(O)R'', -NR'C(O)-OR'', -NR'OR'', in a number ranging from zero to (2m'+1), where m' is the total number of carbon atoms in such radical. R, R', R'', R''', and R'''' each preferably independently refer to hydrogen, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl (e.g., aryl substituted with 1-3 halogens), substituted or unsubstituted heteroaryl, substituted or unsubstituted alkyl, alkoxy, or thioalkoxy groups, or arylalkyl groups. When a compound described herein includes more than one R group, for example, each of the R groups is independently selected as are each R', R'', R''', and R'''' group when more than one of these groups is present. When R' and R'' are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a 4-, 5-, 6-, or 7-membered ring. For example, -NR'R'' includes, but is not limited to, 1-pyrrolidinyl and 4-morpholinyl. From the above discussion of substituents, one of skill in the art will understand that the term “alkyl” is meant to include groups including carbon atoms bound to groups other than hydrogen groups, such as haloalkyl (e.g., -CF3 and -CH2CF3) and acyl (e.g., -C(O)CH3, -C(O)CF3, -C(O)CH2OCH3, and the like).

[0044] Similar to the substituents described for the alkyl radical, substituents for the aryl and heteroaryl groups are varied and are selected from, for example: -OR', -NR'R'', -SR', -halogen, -SiR'R''R''', -OC(O)R', -C(O)R', -CO2R', -CONR'R'', -OC(O)NR'R'', -NR''C(O)R', -NR'- C(O)NR''R''', -NR''C(O)2R', -NR-C(NR'R''R''')=NR'''', -NR-C(NR'R'')=NR''', -S(O)R', -S(O)2R', -S(O)2NR'R'', -NRSO2R', ^NR'NR''R''', ^ONR'R'', ^NR'C(O)NR''NR'''R'''', -CN, -NO2, -R', -N3, -CH(Ph)2, fluoro(C1-C4)alkoxy, and fluoro(C1-C4)alkyl, -NR'SO2R'', -NR'C(O)R'', -NR'C(O)-OR'', -NR'OR'', in a number ranging from zero to the total number of open valences on the aromatic ring system; and where R', R'', R''', and R'''' are preferably independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl. When a compound described herein includes more than one R group, for example, each of the R groups is independently selected as are each R', R'', R''', and R'''' groups when more than one of these groups is present.

[0045] Substituents for rings (e.g. cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene) may be depicted as substituents on the ring rather than on a specific atom of a ring (commonly referred to as a floating substituent). In such a case, the substituent may be attached to any of the ring atoms (obeying the rules of chemical valency) and in the case of fused rings, bridged rings, or spirocyclic rings, a substituent depicted as associated with one member of the fused rings, bridged rings, orspirocyclic rings (a floating substituent on a single ring), may be a substituent on any of the fused rings, bridged rings, or spirocyclic rings (a floating substituent on multiple rings). When a substituent is attached to a ring, but not a specific atom (a floating substituent), and a subscript for the substituent is an integer greater than one, the multiple substituents may be on the same atom, same ring, different atoms, different fused rings, different bridged rings, or different spirocyclic rings, and each substituent may optionally be different. Where a point of attachment of a ring to the remainder of a molecule is not limited to a single atom (a floating substituent), the attachment point may be any atom of the ring and in the case of fused rings, bridged rings, or spirocyclic rings, any atom of any of the fused rings, bridged rings, or spirocyclic rings while obeying the rules of chemical valency. Where a ring, fused rings, bridged rings, or spirocyclic rings contain one or more ring heteroatoms and the ring, fused rings, bridged rings, or spirocyclic rings are shown with one or more floating substituents (including, but not limited to, points of attachment to the remainder of the molecule), the floating substituents may be bonded to the heteroatoms. Where the ring heteroatoms are shown bound to one or more hydrogens (e.g. a ring nitrogen with two bonds to ring atoms and a third bond to a hydrogen) in the structure or formula with the floating substituent, when the heteroatom is bonded to the floating substituent, the substituent will be understood to replace the hydrogen, while obeying the rules of chemical valency.

[0046] Two or more substituents may optionally be joined to form aryl, heteroaryl, cycloalkyl, or heterocycloalkyl groups. Such so-called ring-forming substituents are typically, though not necessarily, found attached to a cyclic base structure. In one embodiment, the ring-forming substituents are attached to adjacent members of the base structure. For example, two ring- forming substituents attached to adjacent members of a cyclic base structure create a fused ring structure. In another embodiment, the ring-forming substituents are attached to a single member of the base structure. For example, two ring-forming substituents attached to a single member of a cyclic base structure create a spirocyclic structure. In yet another embodiment, the ring- forming substituents are attached to non-adjacent members of the base structure and form a bridged ring structure.

[0047] Two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally form a ring of the formula -T-C(O)-(CRR')q-U-, wherein T and U are independently -NR-, -O-, -CRR'-, or a single bond, and q is an integer of from 0 to 3. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula -A-(CH2)r-B-, wherein A and B are independently -CRR'-, -O-, -NR-, -S-, -S(O) -, -S(O)2-, -S(O)2NR'-, or a single bond, and r is an integer of from 1 to 4. One of the single bondsof the new ring so formed may optionally be replaced with a double bond. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula -(CRR')s-X'- (C''R''R''')d-, where s and d are independently integers of from 0 to 3, and X' is -O-, -NR'-, -S-, -S(O)-, -S(O)2-, or -S(O)2NR'-. The substituents R, R', R'', and R''' are preferably independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl.

[0048] The terms “heteroatom” or “ring heteroatom” are meant to include oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), and silicon (Si).

[0049] A “substituent group,” as used herein, means a group selected from the following moieties:

[0050] (A) oxo, halogen, -CCl3, -CBr3, -CF3, -CI3,-CN, -OH, -NH2, -COOH, -CONH2, -NO2, -SH, -SO3H, -SO4H, -SO2NH2, ^NHNH2, ^ONH2, ^NHC(O)NHNH2, -NHC(O)NH2, -NHSO2H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl3, -OCF3, -OCBr3, -OCI3,-OCHCl2, -OCHBr2, -OCHI2, -OCHF2, unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl), unsubstituted heteroalkyl (e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl), unsubstituted aryl (e.g., C6-C10 aryl, C10 aryl, or phenyl), or unsubstituted heteroaryl (e.g., 5 to 10 membered heteroaryl, 5 to 9 membered heteroaryl, or 5 to 6 membered heteroaryl), and

[0051] (B) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, substituted with at least one substituent selected from:

[0052] (i) oxo, halogen, -CCl3, -CBr3, -CF3, -CI3,-CN, -OH, -NH2, -COOH, -CONH2, -NO2, -SH, -SO3H, -SO4H, -SO2NH2, ^NHNH2, ^ONH2, ^NHC(O)NHNH2,^NHC(O)NH2, -NHSO2H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl3, -OCF3, -OCBr3, -OCI3,-OCHCl2, -OCHBr2, -OCHI2, -OCHF2, unsubstituted alkyl (e.g., C1-C8alkyl, C1-C6alkyl, or C1-C4alkyl), unsubstituted heteroalkyl (e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C3-C8cycloalkyl, C3-C6cycloalkyl, or C5-C6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl), unsubstituted aryl (e.g., C6-C10 aryl, C10 aryl, or phenyl), or unsubstituted heteroaryl (e.g., 5 to 10 membered heteroaryl,5 to 9 membered heteroaryl, or 5 to 6 membered heteroaryl), and

[0053] (ii) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, substituted with at least one substituent selected from:

[0054] (a) oxo, halogen, -CCl3, -CBr3, -CF3, -CI3,-CN, -OH, -NH2, -COOH, -CONH2, -NO2, -SH, -SO3H, -SO4H, -SO2NH2, -NHNH2, -ONH2, -NHC(O)NHNH2, -NHSO2H, -NHC(O)NH2, -NHSO2H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl3, -OCF3, -OCBr3, -OCI3,-OCHCl2, -OCHBr2, -OCHI2, -OCHF2, unsubstituted alkyl (e.g., C1-C8 alkyl, C1- C6alkyl, or C1-C4alkyl), unsubstituted heteroalkyl (e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6cycloalkyl, or C5-C6cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl), unsubstituted aryl (e.g., C6-C10aryl, C10aryl, or phenyl), or unsubstituted heteroaryl (e.g., 5 to 10 membered heteroaryl, 5 to 9 membered heteroaryl, or 5 to 6 membered heteroaryl), and

[0055] (b) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, substituted with at least one substituent selected from: oxo, halogen, -CCl3, -CBr3, -CF3, -CI3,-CN, -OH, -NH2, -COOH, -CONH2, -NO2, -SH, -SO3H, -SO4H, -SO2NH2, ^NHNH2, ^ONH2, ^NHC(O)NHNH2, ^NHC(O)NH2, -NHSO2H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl3, -OCF3, -OCBr3, -OCI3,-OCHCl2, -OCHBr2, -OCHI2, -OCHF2, unsubstituted alkyl (e.g., C1-C8alkyl, C1-C6alkyl, or C1-C4 alkyl), unsubstituted heteroalkyl (e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C3-C8cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl), unsubstituted aryl (e.g., C6-C10 aryl, C10 aryl, or phenyl), or unsubstituted heteroaryl (e.g., 5 to 10 membered heteroaryl, 5 to 9 membered heteroaryl, or 5 to 6 membered heteroaryl).

[0056] In embodiments, each substituted group described in the compounds herein is substituted with at least one substituent group. In embodiments, each substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and / or substituted heteroarylene described in the compounds herein are substituted with at least one substituent group. In embodiments, at least one or all of these groups are substituted with at least one size- limited substituent group. In embodiments, at least one or all of these groups are substituted withat least one lower substituent group.

[0057] In embodiments, each substituted or unsubstituted alkyl may be a substituted or unsubstituted C1-C20alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 20 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C8cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 8 membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted C6-C10aryl, and / or each substituted or unsubstituted heteroaryl is a substituted or unsubstituted 5 to 10 membered heteroaryl. In aspects of the compounds herein, each substituted or unsubstituted alkylene is a substituted or unsubstituted C1-C20 alkylene, each substituted or unsubstituted heteroalkylene is a substituted or unsubstituted 2 to 20 membered heteroalkylene, each substituted or unsubstituted cycloalkylene is a substituted or unsubstituted C3-C8 cycloalkylene, each substituted or unsubstituted heterocycloalkylene is a substituted or unsubstituted 3 to 8 membered heterocycloalkylene, each substituted or unsubstituted arylene is a substituted or unsubstituted C6-C10 arylene, and / or each substituted or unsubstituted heteroarylene is a substituted or unsubstituted 5 to 10 membered heteroarylene.

[0058] In embodiments, each substituted or unsubstituted alkyl is a substituted or unsubstituted C1-C8alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C7cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 7 membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted C6-C10aryl, and / or each substituted or unsubstituted heteroaryl is a substituted or unsubstituted 5 to 9 membered heteroaryl. In embodiments, each substituted or unsubstituted alkylene is a substituted or unsubstituted C1-C8alkylene, each substituted or unsubstituted heteroalkylene is a substituted or unsubstituted 2 to 8 membered heteroalkylene, each substituted or unsubstituted cycloalkylene is a substituted or unsubstituted C3-C7 cycloalkylene, each substituted or unsubstituted heterocycloalkylene is a substituted or unsubstituted 3 to 7 membered heterocycloalkylene, each substituted or unsubstituted arylene is a substituted or unsubstituted C6-C10 arylene, and / or each substituted or unsubstituted heteroarylene is a substituted or unsubstituted 5 to 9 membered heteroarylene.

[0059] Certain compounds may possess asymmetric carbon atoms (optical or chiral centers) or double bonds; the enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisometric forms that may be defined, in terms of absolute stereochemistry, as (R)-or (S)- or, as (D)- or (L)- for amino acids, and individual isomers are encompassed herein. Thecompounds do not include those that are known in art to be too unstable to synthesize and / or isolate. The disclosure is meant to include compounds in racemic and optically pure forms. Optically active (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When the compounds described herein contain olefinic bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers. Unless otherwise stated, structures depicted herein are also meant to include all stereochemical forms of the structure; i.e., the R and S configurations for each asymmetric center. Therefore, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the compounds are within the scope of the disclosure. The term “isomers” refers to compounds having the same number and kind of atoms, and hence the same molecular weight, but differing in respect to the structural arrangement or configuration of the atoms. The term “tautomer” refers to one of two or more structural isomers which exist in equilibrium and which are readily converted from one isomeric form to another. It will be apparent to one skilled in the art that certain compounds may exist in tautomeric forms, all such tautomeric forms of the compounds being within the scope of the disclosure.

[0060] Unless otherwise stated, structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by13C- or14C-enriched carbon are within the scope of this disclosure. The compounds may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may be radiolabeled with radioactive isotopes, such as for example tritium (3H), iodine-125 (125I), or carbon-14 (14C). All isotopic variations of the compounds, whether radioactive or not, are encompassed herein.

[0061] It should be noted that throughout the application that alternatives are written in Markush groups, for example, each amino acid position that contains more than one possible amino acid. It is specifically contemplated that each member of the Markush group should be considered separately, thereby comprising another embodiment, and the Markush group is not to be read as a single unit.

[0062] The terms “a” or “an,” as used in herein means one or more. In addition, the phrase “substituted with a[n],” as used herein, means the specified group may be substituted with one or more of any or all of the named substituents. For example, where a group, such as an alkyl or heteroaryl group, is “substituted with an unsubstituted C1-C20 alkyl, or unsubstituted 2 to 20membered heteroalkyl,” the group may contain one or more unsubstituted C1-C20alkyls, and / or one or more unsubstituted 2 to 20 membered heteroalkyls.

[0063] Where a moiety is substituted with an R substituent, the group may be referred to as “R-substituted.” Where a moiety is R-substituted, the moiety is substituted with at least one R substituent and each R substituent is optionally different. Where a particular R group is present in the description of a chemical genus (such as Formula (I)), a Roman alphabetic symbol may be used to distinguish each appearance of that particular R group. For example, where multiple R13substituents are present, each R1substituent may be distinguished as R1A, R1B, etc., wherein each of R1A, R1B, etc. is defined within the scope of the definition of R1and optionally differently.

[0064] A “covalent cysteine modifier moiety” as used herein refers to a substituent that is capable of reacting with the sulfhydryl functional group of a cysteine amino acid (e.g. cysteine 442 of the interleukin-2-inducible T-cell kinase), or amino acid corresponding to cysteine 442 of the interleukin-2-inducible T-cell kinase) to form a covalent bond. Thus, the covalent cysteine modifier moiety is typically electrophilic. The term “electrophilic chemical moiety” is used in accordance with its plain ordinary chemical meaning and refers to a monovalent chemical group that is electrophilic.

[0065] Descriptions of compounds are limited by principles of chemical bonding known to those skilled in the art. Accordingly, where a group may be substituted by one or more of a number of substituents, such substitutions are selected so as to comply with principles of chemical bonding and to give compounds which are not inherently unstable and / or would be known to one of ordinary skill in the art as likely to be unstable under ambient conditions, such as aqueous, neutral, and several known physiological conditions. For example, a heterocycloalkyl or heteroaryl is attached to the remainder of the molecule via a ring heteroatom in compliance with principles of chemical bonding known to those skilled in the art thereby avoiding inherently unstable compounds.

[0066] “T cell factor-1” or “Tcf1” refer to a protein (including homologs, isoforms, and functional fragments thereof) with Tcf1 activity. The term includes any recombinant or naturally-occurring form of Tcf1 or isoforms or variants thereof that maintain Tcf1 activity (e.g.within at least 60%, 70%, 80%, 90%, or 100% activity compared to wildtype Tcf1). TCF1 isrequired for the self-renewal of stem-like CD8+T cells generated in response to viral or tumour antigens, and for preserving heightened responses to checkpoint blockade immunotherapy. In the helper T cell lineages, TCF1 is indispensable for the differentiation of T follicular helper and T follicular regulatory cells, and crucially regulates immunosuppressive functions of regulatory T cells. In embodiments, the Tcf1 protein is encoded by the transcription factor 7 (TCF7) gene.In aspects, the TCF7 gene has the nucleic acid sequence set forth in NCBI Gene ID No.6932. Tcf1 plays an important role during T-cell development and differentiation for T-cells to exercise their functions, including producing memory T-cells. Tcf1 can modulate T-cell development, exert various effects on the differentiation and function of mature CD8+ T-cells, and drive the production and maintenance of the immune response of CD8+T cells (e.g., after PD-1 checkpoint blockade therapy).

[0067] The term “LAG3” or “lymphocyte activation gene 3 protein” refer to a protein (including homologs, isoforms, and functional fragments thereof) with LAG3 activity. The term includes any recombinant or naturally-occurring form of LAG3 or variants thereof that maintain LAG3 activity (e.g. within at least 60%, 70%, 80%, 90%, or 100% activity compared to wildtype LAG3). In aspects, the LAG3 protein has the amino acid sequence set forth in or corresponding to UniProt P18627. In aspects, the LAG3 gene has the nucleic acid sequence set forth in NCBI Gene ID No.3902.

[0068] The term “TIGIT” or “T-cell immunoreceptor with Ig and ITIM domains” refer to a protein (including homologs, isoforms, and functional fragments thereof) with TIGIT activity. The term includes any recombinant or naturally-occurring form of TIGIT or variants thereof that maintain TIGIT activity (e.g. within at least 60%, 70%, 80%, 90%, or 100% activity compared to wildtype TIGIT). In aspects, the TIGIT protein has the amino acid sequence set forth in or corresponding to UniProt Q495A1. In aspects, the TIGIT gene has the nucleic acid sequence set forth in NCBI Gene ID No.201633.

[0069] The term “PD-1” or “programmed cell death protein 1” refer to a protein (including homologs, isoforms, and functional fragments thereof) with PD-1 activity. The term includes any recombinant or naturally-occurring form of PD-1 or variants thereof that maintain PD-1 activity (e.g. within at least 60%, 70%, 80%, 90%, or 100% activity compared to wildtype PD- 1). In aspects, the PD-1 protein encoded by the PDCD1 gene has the amino acid sequence set forth in or corresponding to UniProt Q15116. In aspects, the PDCD1 gene has the nucleic acid sequence set forth in NCBI Gene ID No.5133.

[0070] The term “IFNγ” or “interferon gamma” refer to a protein (including homologs, isoforms, and functional fragments thereof) with IFNγ activity. The term includes any recombinant or naturally-occurring form of IFNγ or variants thereof that maintain IFNγ activity (e.g. within at least 60%, 70%, 80%, 90%, or 100% activity compared to wildtype IFNγ). In aspects, the IFNγ protein encoded by the IFNG gene has the amino acid sequence set forth in or corresponding to UniProt P01579.

[0071] The term “granzyme B” refers to a protein (including homologs, isoforms, and functional fragments thereof) with granzyme B activity. The term includes any recombinant or naturally-occurring form of granzyme B or variants thereof that maintain granzyme B activity (e.g. within at least 60%, 70%, 80%, 90%, or 100% activity compared to wildtype granzyme B). In aspects, the granzyme B protein encoded by the GZMB gene has the amino acid sequence set forth in or corresponding to UniProt J3KPK2.

[0072] The term “TIM3” or “T-cell immunoglobulin mucin receptor 3” refer to a protein (including homologs, isoforms, and functional fragments thereof) with TIM3 activity. The term includes any recombinant or naturally-occurring form of TIM3 or variants thereof that maintain TIM3 activity (e.g. within at least 60%, 70%, 80%, 90%, or 100% activity compared to wildtype TIM3). In aspects, the TIM3 protein encoded by the HAVCR2 gene has the amino acid sequence set forth in or corresponding to UniProt Q8TDQ0. An alternative name for TIM3 is hepatitis A virus cellular receptor 2 or HAVcr-2.

[0073] “Treating” or “treatment” as used herein (and as well-understood in the art) broadly includes any approach for obtaining beneficial or desired results in a subject’s condition, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of the extent of a disease, stabilizing (i.e., not worsening) the state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission, whether partial or total and whether detectable or undetectable. The term “treating” does not include “preventing.”

[0074] “Biological sample” or “sample” refer to materials obtained from or derived from a subject or patient. A biological sample includes sections of tissues such as biopsy. Such samples include bodily fluids such as blood and blood fractions or products (e.g., serum, plasma, platelets, red blood cells, and the like), sputum, tissue, cultured cells (e.g., primary cultures, explants, and transformed cells), stool, urine, and the like. In embodiments, a biological sample is blood. In embodiments, a biological sample is a tumor cell. In embodiments, a biological sample is a tumor.

[0075] The term “biomarker” refers to an indicator, e.g., a predictive, prognostic, and / or a pharmacodynamic indicator, which can be detected in a biological sample. The biomarker may serve as an indicator of the likelihood a patient will respond to a particular therapeutic treatment or a particular subtype of a disease, characterized by certain molecular, pathological, histological, and / or clinical features. In embodiments, a biomarker is a gene or a set of genes (i.e., a biomarker gene). Biomarkers include, but are not limited to, polynucleotides (e.g., DNA,and / or RNA), polynucleotide copy number alterations (e.g., DNA copy numbers), polypeptides, or polypeptide and polynucleotide modifications (e.g., posttranslational modifications). In embodiments, the biomarker is LAG3, TIGIT, PD-1, IFNγ, or granzyme B.

[0076] The terms an “increased expression level” or “increased level” of gene expression is an expression level of the gene that is higher than the expression level of the gene in a control. The control may be any control known in the art, such as those described herein. In embodiments, an “increased level” of the biomarker gene compared to the control (when the expression level of the biomarker is greater than the corresponding control) is, for example, an increase in the expression level of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98% or 99% or greater relative to the control. In embodiments, an “increased level” of the biomarker gene is an amount that is statistically significantly greater than the expression level of the control.

[0077] The terms a “decreased expression level” or “decreased level” of gene expression is an expression level of the gene that is lower than the expression level of the gene in a control. The control may be any control known in the art, such as those described herein. In embodiments, a “decreased level” of the biomarker gene compared to the control (when the expression level of the biomarker is lower than the corresponding control) is, for example, a decrease in the expression level of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98% or 99% or greater relative to the control. In embodiments, a “decreased level” of the biomarker gene is an amount that is statistically significantly lower than the expression level of the control.

[0078] “Control” is used in accordance with its plain ordinary meaning and refers to an assay, comparison, or experiment in which the subjects or reagents of the experiment are treated as in a parallel experiment except for omission of a procedure, reagent, or variable of the experiment. In embodiments, the control is used as a standard of comparison in evaluating experimental effects. In embodiments, the control is a gene expression level against which another gene expression level (e.g. the gene expression level of a biomarker gene disclosed herein) is compared (e.g., to make a diagnostic (e.g., predictive and / or prognostic) and / or therapeutic determination. In embodiments, a control is a healthy patient or a healthy population of patients. In embodiments, a healthy patient is a patient that does not have cancer. In embodiments, the control is an average value from population of healthy patients.

[0079] Biomarker levels may be detected at either the protein (e.g., cytokine) or gene expression level. Proteins expressed by biomarkers can be quantified by immunohistochemistry (IHC), ELISA, or flow cytometry with an antibody that detects the proteins. Biomarkerexpression can be and quantified by multiple platforms known in the art. Quantifying biomarker (gene) expression can alternatively be referred to as detecting a level of biomarker (gene) expression. Platforms that can be used to quantify biomarker (gene) expression or detect levels of biomarker (gene) expression include quantitative polymerase chain reaction (qPCR), multiplex quantitative polymerase chain reaction (multiplex qPCR), real-time polymerase chain reaction (rtPCR), Nanostring (e.g., an amplification-free technology that measures nucleic acid content by counting molecules directly), RNA-sequencing (using next-generation sequencing (NGS) to reveal the presence and quantity of RNA in a biological sample), or in situ hybridization. There is a range of biomarker expression across as measured by Nanostring. In embodiments, quantitative rtPCR, Nanostring, RNA-sequencing (RNAseq), and in situ hybridization are used to quantitate biomarker gene expression. In embodiments, biomarker expression is quantified by RNAseq. In embodiments, biomarker expression is quantified by multiplex qPCR. In embodiments, biomarker expression is quantified by NanoString. For Nanostring, RNA is extracted from a biological sample and a known quantity of RNA is placed on the Nanostring machine for gene expression detection using gene specific probes. The number of counts of biomarkers within a sample is determined and normalized to a set of housekeeping genes. In embodiments, the increased or decreased expression of biomarkers may be determined by calculating the H-score for the expression of the biomarkers.

[0080] “Contacting” is used in accordance with its plain ordinary meaning and refers to the process of allowing at least two distinct species (e.g. chemical compounds including biomolecules or cells) to become sufficiently proximal to react, interact or physically touch. In embodiments contacting includes allowing an ITK inhibitor described herein to interact with a T-cell or a CAR T-cell.

[0081] Methods

[0082] Provided herein are methods for making CAR T-cells comprising contacting an ITK inhibitor with CAR T-cells in vitro. Provided herein are methods for increasing expression of T- cell factor-1 on CAR T-cells comprising contacting an ITK inhibitor with CAR T-cells in vitro, thereby increasing expression of T-cell factor-1 on CAR T-cells. Provided herein are methods for increasing CAR T-cell function comprising contacting an ITK inhibitor with CAR T-cells in vitro, thereby increasing CAR T-cell function. In embodiments, the CAR T-cell function is CAR T-cell cytolytic function. Provided herein are methods of reducing CAR T-cell exhaustion comprising contacting an ITK inhibitor with CAR T-cells in vitro, thereby reducing CAR T-cell exhaustion. In embodiments, the CAR T-cells are CD4+ T-cells. In embodiments, the CAR T- cells are CD8+ T-cells. In embodiments, the CAR T-cells are CD4+ T-cells and CD8+ T-cells.In embodiments, the CAR T-cells are autologous. In embodiments, the CAR T-cells are allogeneic. The ITK inhibitor is contacted with the CAR T-cells by preparing a composition comprising the ITK inhibitor and the CAR T-cells. In embodiments, the composition comprises an ITK inhibitor and CAR T-cells. In embodiments, the composition comprises an ITK inhibitor, CAR T-cells, and serum. In embodiments, the composition comprises an ITK inhibitor, CAR T-cells, and human serum. In embodiments, the composition comprises an ITK inhibitor, CAR T-cells, and fetal bovine serum. In embodiments, the composition comprises an ITK inhibitor, CAR T-cells, and a serum-free medium. In embodiments, contacting an ITK inhibitor with CAR T-cells in vitro results in ITK-contacted CAR T-cells. ITK-contacted CAR T-cells have improved properties relative to CAR T-cells that have not been contacted with an ITK inhibitor. In embodiments, the ITK-contacted CAR T-cells have an increased expression of Tcf1 when compared to CAR T-cells that have not been contacted with an ITK inhibitor. In embodiments, the ITK-contacted CAR T-cells have increased CAR T-cell function when compared to CAR T-cells that have not been contacted with an ITK inhibitor. In embodiments, the ITK-contacted CAR T-cells have increased cytolytic function when compared to CAR T- cells that have not been contacted with an ITK inhibitor. In embodiments, the ITK-contacted CAR T-cells have a decreased level of LAG3, a decreased level of TIGIT, a decreased level of PD-1, an increased level of IFNγ, an increased level of granzyme B, a decreased level of TIM3, or a combination of two or more thereof when compared to CAR T-cells that have not been contacted with an ITK inhibitor. In embodiments, the CAR T-cells that have not been contacted with an ITK inhibitor are prepared from autologous T cells.

[0083] Provided herein are methods of treating cancer in a patient in need thereof comprising administering to the patient: (i) an effective amount of an ITK inhibitor and (ii) an effective amount of CAR T-cells, thereby treating cancer in the patient. Provided herein are methods of treating cancer in a patient in need thereof comprising: (i) contacting CAR T-cells with an effective amount of an ITK inhibitor in vitro, thereby producing ITK-contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK-contacted CAR T-cells; thereby treating cancer in the patient. In embodiments, the method further comprises administering to the patient an effective amount of the ITK inhibitor. In embodiments, the CAR T-cells are CD4+ T-cells. In embodiments, the CAR T-cells are CD8+ T-cells. In embodiments, the CAR T-cells are CD4+ T-cells and CD8+ T-cells. In embodiments, the CAR T-cells are autologous. In embodiments, the CAR T-cells are allogeneic. In embodiments, the ITK inhibitor is any ITK inhibitor described herein (including embodiments thereof). In embodiments, the ITK inhibitor is soquelitinib or a pharmaceutically acceptable salt thereof. In embodiments, theCAR T-cells are any CAR T-cells known in the art or described herein (including embodiments thereof). In embodiments, the cancer is any cancer known in the art and described herein (including embodiments thereof).

[0084] Provided herein are methods for preventing and / or reversing T-cell exhaustion in a patient in need thereof, the method comprising administering to the patient an effective amount of an ITK inhibitor; thereby preventing and / or reversing T-cell exhaustion in the patient. In embodiments, the T-cell is a CD4+ T-cell, a CD8+ T-cell, or a combination thereof. In embodiments, the T-cell is a CAR T-cell. Provided herein are methods for preventing and / or reversing CAR T-cell exhaustion in a patient in need thereof comprising administering to the patient an effective amount of an ITK inhibitor; wherein the patient is receiving or has received CAR T-cell therapy; thereby preventing and / or reversing CAR T-cell exhaustion in the patient. Provided herein are methods of preventing and / or reversing CAR T-cell exhaustion in a patient in need thereof comprising administering to the patient: (i) an effective amount of an ITK inhibitor and (ii) an effective amount of CAR T-cells, thereby preventing and / or reducing CAR T-cell exhaustion in the patient. Provided herein are methods of preventing and / or reversing CAR T-cell exhaustion in a patient in need thereof comprising: (i) contacting CAR T-cells with an effective amount of an ITK inhibitor in vitro, thereby producing ITK-contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK-contacted CAR T-cells; thereby preventing and / or reducing CAR T-cell exhaustion in the patient. In embodiments, the method further comprises administering to the patient an effective amount of the ITK inhibitor. In embodiments, the CAR T-cells are CD4+ T-cells. In embodiments, the CAR T-cells are CD8+ T-cells. In embodiments, the CAR T-cells are CD4+ T-cells and CD8+ T-cells. In embodiments, the CAR T-cells are autologous. In embodiments, the CAR T-cells are allogeneic. In embodiments, the ITK inhibitor is any ITK inhibitor described herein (including embodiments thereof). In embodiments, the ITK inhibitor is soquelitinib or a pharmaceutically acceptable salt thereof. In embodiments, the CAR T-cells are any CAR T-cells known in the art or described herein (including embodiments thereof). In embodiments, the patient has cancer. In embodiments, the methods comprise treating cancer in the patient. In embodiments, the cancer is any cancer known in the art and described herein (including embodiments thereof).

[0085] Provided herein are methods for stimulating a T-cell-mediated immune response in a patient in need thereof comprising administering to the patient an effective amount of an ITK inhibitor; thereby stimulating the T-cell-mediated immune response in the patient. In embodiments, the T-cell is a CD4+ T-cell, a CD8+ T-cell, or a combination thereof. In embodiments, the T-cells are CAR T-cells. Provided herein are methods for stimulating a CART-cell-mediated immune response in a patient in need thereof comprising administering to the patient an effective amount of an ITK inhibitor; wherein the patient is receiving or has received CAR T-cell therapy; thereby stimulating the CAR T-cell-mediated immune response in the patient. Provided herein are methods for stimulating a CAR T-cell-mediated immune response in a patient in need thereof comprising administering to the patient: (i) an effective amount of an ITK inhibitor and (ii) an effective amount of CAR T-cells, thereby stimulating the CAR T-cell- mediated immune response in the patient. Provided herein are methods for stimulating a CAR T- cell-mediated immune response in a patient in need thereof comprising: (i) contacting CAR T- cells with an effective amount of an ITK inhibitor in vitro, thereby producing ITK-contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK-contacted CAR T-cells; thereby stimulating the CAR T-cell-mediated immune response in the patient. In embodiments, the methods further comprise administering to the patient an effective amount of the ITK inhibitor. In embodiments, the CAR T-cells are CD4+ T-cells. In embodiments, the CAR T-cells are CD8+ T-cells. In embodiments, the CAR T-cells are CD4+ T-cells and CD8+ T-cells. In embodiments, the CAR T-cells are autologous. In embodiments, the CAR T-cells are allogeneic. In embodiments, the ITK inhibitor is any ITK inhibitor described herein (including embodiments thereof). In embodiments, the ITK inhibitor is soquelitinib or a pharmaceutically acceptable salt thereof. In embodiments, the CAR T-cells are any CAR T-cells known in the art or described herein (including embodiments thereof). In embodiments, the patient has cancer. In embodiments, the methods comprise treating cancer in the patient. In embodiments, the cancer is any cancer known in the art and described herein (including embodiments thereof).

[0086] Provided herein are methods for increasing T-cell function in a patient in need thereof comprising administering to the patient an effective amount of an i ITK inhibitor; thereby increasing T-cell function in the patient. In embodiments, the T-cell is a CD4+ T-cell, a CD8+ T-cell, or a combination thereof. In embodiments, the T-cell is a CAR T-cell. Provided herein are methods for increasing CAR T-cell function in a patient in need thereof comprising administering to the patient an effective amount of the ITK inhibitor; wherein the patient is receiving or has received CAR T-cell therapy; thereby increasing CAR T-cell function in the patient. Provided herein are methods for increasing CAR T-cell function in a patient in need thereof comprising administering to the patient: (i) an effective amount of an ITK inhibitor and (ii) an effective amount of CAR T-cells, thereby increasing CAR T-cell function in the patient. Provided herein are methods for increasing CAR T-cell function in a patient in need thereof comprising: (i) contacting CAR T-cells with an effective amount of an ITK inhibitor in vitro, thereby producing ITK-contacted CAR T-cells; and (ii) administering to the patient an effectiveamount of the ITK-contacted CAR T-cells; thereby increasing CAR T-cell function in the patient. In embodiments, the methods further comprise administering to the patient an effective amount of the ITK inhibitor. In embodiments, the T-cell function is T-cell cytolytic function. In embodiments, the T-cell function is expression of Tcf1. In embodiments, the CAR T-cells are CD4+ T-cells. In embodiments, the CAR T-cells are CD8+ T-cells. In embodiments, the CAR T-cells are CD4+ T-cells and CD8+ T-cells. In embodiments, the CAR T-cells are autologous. In embodiments, the CAR T-cells are allogeneic. In embodiments, the ITK inhibitor is any ITK inhibitor described herein (including embodiments thereof). In embodiments, the ITK inhibitor is soquelitinib or a pharmaceutically acceptable salt thereof. In embodiments, the CAR T-cells are any CAR T-cells known in the art or described herein (including embodiments thereof). In embodiments, the patient has cancer. In embodiments, the methods comprise treating cancer in the patient. In embodiments, the cancer is any cancer known in the art and described herein (including embodiments thereof).

[0087] Provided herein are methods of increasing expression of Tcf1 on T-cells in a patient in need thereof, the method comprising administering to the patient an effective amount of an ITK inhibitor; thereby increasing expression of Tcf1 on T-cells in the patient. In embodiments, the T- cell is a CD4+ T-cell, a CD8+ T-cell, or a combination thereof. In embodiments, the T-cells are CAR T-cells. Provided herein are methods for increasing expression of Tcf1 on CAR T-cells in a patient in need thereof, the method comprising administering to the patient an effective amount of an ITK inhibitor; wherein the patient is receiving or has received CAR T-cell therapy; thereby increasing expression of Tcf1 on CAR T-cells in the patient. Provided herein are methods of increasing expression of Tcf1 on CAR T-cells in a patient in need thereof, the method comprising administering to the patient (i) an effective amount of an ITK inhibitor and (ii) an effective amount of CAR T-cells, thereby increasing expression of Tcf1 on CAR T-cells in the patient. Provided herein are methods for increasing expression of T-cell factor-1 on CAR T-cells in a patient in need thereof, the method comprising: (i) contacting CAR T-cells with an effective amount of an ITK inhibitor in vitro, thereby producing ITK-contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK-contacted CAR T-cells; thereby increasing expression of T-cell factor-1 on CAR T-cells in the patient. In embodiments, the methods further comprise administering to the patient an effective amount of the ITK inhibitor. In embodiments, the CAR T-cells are CD4+ T-cells. In embodiments, the CAR T-cells are CD8+ T-cells. In embodiments, the CAR T-cells are CD4+ T-cells and CD8+ T-cells. In embodiments, the CAR T-cells are autologous. In embodiments, the CAR T-cells are allogeneic. In embodiments, the ITK inhibitor is any ITK inhibitor described herein (includingembodiments thereof). In embodiments, the ITK inhibitor is soquelitinib or a pharmaceutically acceptable salt thereof. In embodiments, the CAR T-cells are any CAR T-cells known in the art or described herein (including embodiments thereof). In embodiments, the patient has cancer. In embodiments, the methods comprise treating cancer in the patient. In embodiments, the cancer is any cancer known in the art and described herein (including embodiments thereof).

[0088] Provided herein are methods of decreasing expression of LAG3 on T-cells, decreasing expression of TIGIT on T-cells, decreasing expression of PD-1 on T-cells, increasing expression of IFNγ on T-cells, increasing expression of granzyme B on T-cells, decreasing expression of TIM3 on T-cells, or a combination of two or more thereof in a patient in need thereof, the method comprising administering to the patient an effective amount of an ITK inhibitor; thereby decreasing expression of LAG3 on T-cells, decreasing expression of TIGIT on T-cells, decreasing expression of PD-1 on T-cells, increasing expression of IFNγ on T-cells, increasing expression of granzyme B on T-cells, decreasing expression of TIM3 on T-cells, or a combination of two or more thereof in the patient. In embodiments, the T-cell is a CD4+ T-cell, a CD8+ T-cell, or a combination thereof. In embodiments, the T-cells are CAR T-cells. Provided herein are methods for decreasing expression of LAG3 on CAR T-cells, decreasing expression of TIGIT on CAR T-cells, decreasing expression of PD-1 on CAR T-cells, increasing expression of IFNγ on CAR T-cells, increasing expression of granzyme B on CAR T-cells, decreasing expression of TIM3 on CAR T-cells, or a combination of two or more thereof in a patient in need thereof, the method comprising administering to the patient an effective amount of an ITK inhibitor; wherein the patient is receiving or has received CAR T-cell therapy; thereby decreasing expression of LAG3 on CAR T-cells, decreasing expression of TIGIT on CAR T- cells, decreasing expression of PD-1 on CAR T-cells, increasing expression of IFNγ on CAR T- cells, increasing expression of granzyme B on CAR T-cells, decreasing expression of TIM3 on CAR T-cells in the patient. Provided herein are methods of decreasing expression of LAG3 on CAR T-cells, decreasing expression of TIGIT on CAR T-cells, decreasing expression of PD-1 on CAR T-cells, increasing expression of IFNγ on CAR T-cells, increasing expression of granzyme B on CAR T-cells, decreasing expression of TIM3 on CAR T-cells in a patient in need thereof, the method comprising administering to the patient (i) an effective amount of an ITK inhibitor and (ii) an effective amount of CAR T-cells, thereby decreasing expression of LAG3 on CAR T-cells, decreasing expression of TIGIT on CAR T-cells, decreasing expression of PD-1 on CAR T-cells, increasing expression of IFNγ on CAR T-cells, increasing expression of granzyme B on CAR T-cells, decreasing expression of TIM3 on CAR T-cells in the patient. Provided herein are methods for decreasing expression of LAG3 on CAR T-cells, decreasingexpression of TIGIT on CAR T-cells, decreasing expression of PD-1 on CAR T-cells, increasing expression of IFNγ on CAR T-cells, increasing expression of granzyme B on CAR T-cells, decreasing expression of TIM3 on CAR T-cells in a patient in need thereof, the method comprising: (i) contacting CAR T-cells with an effective amount of an ITK inhibitor in vitro, thereby producing ITK-contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK-contacted CAR T-cells; thereby decreasing expression of LAG3 on CAR T- cells, decreasing expression of TIGIT on CAR T-cells, decreasing expression of PD-1 on CAR T-cells, increasing expression of IFNγ on CAR T-cells, increasing expression of granzyme B on CAR T-cells, decreasing expression of TIM3 on CAR T-cells in the patient. In embodiments, the methods further comprise administering to the patient an effective amount of the ITK inhibitor. In embodiments, the CAR T-cells are CD4+ T-cells. In embodiments, the CAR T-cells are CD8+ T-cells. In embodiments, the CAR T-cells are CD4+ T-cells and CD8+ T-cells. In embodiments, the CAR T-cells are autologous. In embodiments, the CAR T-cells are allogeneic. In embodiments, the ITK inhibitor is any ITK inhibitor described herein (including embodiments thereof). In embodiments, the ITK inhibitor is soquelitinib or a pharmaceutically acceptable salt thereof. In embodiments, the CAR T-cells are any CAR T-cells known in the art or described herein (including embodiments thereof). In embodiments, the patient has cancer. In embodiments, the methods comprise treating cancer in the patient. In embodiments, the cancer is any cancer known in the art and described herein (including embodiments thereof).

[0089] In embodiments of the methods described herein, the patient is tested for T-cell exhaustion. The testing for T-cell exhaustion can occur at any time the patient is being treated for cancer. In embodiments, the methods further comprise measuring an increased level of TIM3, an increased level of LAG3, an increased level of TIGIT, an increased level of PD-1, a decreased level of IFNγ, a decreased level of granzyme B, or a combination of two or more thereof, relative to a control, in a biological sample obtained from the patient. In embodiments, a biological sample obtained from the patient has an increased level of TIM3, an increased level of LAG3, an increased level of TIGIT, an increased level of PD-1, a decreased level of IFNγ, a decreased level of granzyme B, or a combination of two or more thereof, relative to a control. When the patient has an increased level of LAG3, an increased level of TIGIT, an increased level of PD-1, a decreased level of IFNγ, a decreased level of granzyme B, or a combination of two or more thereof, relative to a control, the patient has T-cell exhaustion.

[0090] In embodiments of the methods described herein, the patient has cancer and is being treated for cancer. In embodiments, the cancer is lymphoma. In embodiments, the cancer is leukemia. In embodiments, the lymphoma is T-cell lymphoma. In embodiments, the lymphomais peripheral T-cell lymphoma. In embodiments, the lymphoma is peripheral T-cell lymphoma not otherwise specified. In embodiments, the lymphoma is cutaneous T-cell lymphoma. In embodiments, the lymphoma is cutaneous T-cell lymphoma not otherwise specified. In embodiments, the lymphoma is angioimmunoblastic T cell lymphoma. In embodiments, the lymphoma is NK T cell lymphoma. In embodiments, the lymphoma is B-cell lymphoma. In embodiments, the lymphoma is diffuse large B-cell lymphoma. In embodiments, the lymphoma is follicular lymphoma. In embodiments, the lymphoma is high grade B-cell lymphoma. In embodiments, the lymphoma is chronic lymphocytic leukemia. In embodiments, the lymphoma is mantle cell lymphoma. In embodiments, the lymphoma is marginal zone lymphoma. In embodiments, the lymphoma is Burkitt lymphoma. In embodiments, the cancer is B-cell precursor acute lymphoblastic leukemia. In embodiments, the lymphoma is lymphoplasmacytic lymphoma. In embodiments, the cancer is multiple myeloma. In embodiments, the cancer is T cell leukemia. In embodiments, the cancer is relapsed / refractory cancer.

[0091] In embodiments, the cancer is a solid tumor. In embodiments, the cancer is lung cancer, colorectal cancer, pancreatic cancer, prostate cancer, breast cancer, gastric cancer, renal cancer, melanoma, or head and neck cancer. In embodiments, the cancer is lung cancer. In embodiments, the cancer is colorectal cancer. In embodiments, the cancer is pancreatic cancer. In embodiments, the cancer is prostate cancer. In embodiments, the cancer is breast cancer. In embodiments, the cancer is gastric cancer. In embodiments, the cancer is renal cancer. In embodiments, the cancer is melanoma. In embodiments, the cancer is head and neck cancer. In embodiments, the cancer is leukemia. In embodiments, the cancer is relapsed / refractory cancer

[0092] The term “lymphoma” refers to a group of cancers affecting hematopoietic and lymphoid tissues. It begins in lymphocytes, the blood cells that are found primarily in lymph nodes, spleen, thymus, and bone marrow. Two main types of lymphoma are non-Hodgkin lymphoma and Hodgkin’s disease. Hodgkin’s disease represents approximately 15% of all diagnosed lymphomas. This is a cancer associated with Reed-Sternberg malignant B lymphocytes. Non-Hodgkin’s lymphomas (NHL) can be classified based on the rate at which cancer grows and the type of cells involved. There are aggressive (high grade) and indolent (low grade) types of NHL. Based on the type of cells involved, there are B-cell and T-cell NHLs. Exemplary B-cell lymphomas that may be treated with a compound or method provided herein include, but are not limited to, small lymphocytic lymphoma, Mantle cell lymphoma, follicular lymphoma, marginal zone lymphoma, extranodal (MALT) lymphoma, nodal (monocytoid B- cell) lymphoma, splenic lymphoma, diffuse large cell B-lymphoma, Burkitt’s lymphoma, lymphoblastic lymphoma, immunoblastic large cell lymphoma, or precursor B-lymphoblasticlymphoma. Exemplary T-cell lymphomas that may be treated with a compound or method provided herein include, but are not limited to, cutaneous T-cell lymphoma, peripheral T-cell lymphoma (including angioimmunoblastic T cell lymphoma and peripheral T cell lymphoma not otherwise specified), anaplastic large cell lymphoma, mycosis fungoides, NK T cell lymphoma and precursor T-lymphoblastic lymphoma..

[0093] The term “leukemia” refers broadly to progressive, malignant diseases of the blood- forming organs and is generally characterized by a distorted proliferation and development of leukocytes and their precursors in the blood and bone marrow. Leukemia is generally clinically classified on the basis of (1) the duration and character of the disease-acute or chronic; (2) the type of cell involved; myeloid (myelogenous), lymphoid (lymphogenous), or monocytic; and (3) the increase or non-increase in the number abnormal cells in the blood-leukemic or aleukemic (subleukemic). Exemplary leukemias that may be treated with a compound or method provided herein include, for example, acute nonlymphocytic leukemia, chronic lymphocytic leukemia, acute granulocytic leukemia, chronic granulocytic leukemia, acute promyelocytic leukemia, adult T-cell leukemia, aleukemic leukemia, a leukocythemic leukemia, basophylic leukemia, blast cell leukemia, bovine leukemia, chronic myelocytic leukemia, leukemia cutis, embryonal leukemia, eosinophilic leukemia, Gross' leukemia, hairy-cell leukemia, hemoblastic leukemia, hemocytoblastic leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia, leukopenic leukemia, lymphatic leukemia, lymphoblastic leukemia, lymphocytic leukemia, lymphogenous leukemia, lymphoid leukemia, lymphosarcoma cell leukemia, mast cell leukemia, megakaryocytic leukemia, micromyeloblastic leukemia, monocytic leukemia, myeloblastic leukemia, myelocytic leukemia, myeloid granulocytic leukemia, myelomonocytic leukemia, Naegeli leukemia, plasma cell leukemia, multiple myeloma, plasmacytic leukemia, promyelocytic leukemia, Rieder cell leukemia, Schilling's leukemia, stem cell leukemia, subleukemic leukemia, or undifferentiated cell leukemia.

[0094] ITK Inhibitors

[0095] In embodiments, the ITK inhibitor is a compound having a selectivity for ITK that is at least 50-fold greater than the selectivity for resting lymphocyte kinase (RLK). In embodiments, the ITK inhibitor is a compound having a selectivity for ITK that is at least 60-fold greater than the selectivity for RLK. In embodiments, the ITK inhibitor is a compound having a selectivity for ITK that is at least 70-fold greater than the selectivity for RLK. In embodiments, the ITK inhibitor is a compound having a selectivity for ITK that is at least 80-fold greater than the selectivity for RLK. In embodiments, the ITK inhibitor is a compound having a selectivity for ITK that is at least 90-fold greater than the selectivity for RLK. In embodiments, the ITKinhibitor is a compound having a selectivity for ITK that is at least 100-fold greater than the selectivity for RLK. In embodiments, the ITK inhibitor is a compound having a selectivity for ITK that is at least 110-fold greater than the selectivity for RLK.

[0096] In embodiments, the ITK inhibitor is a compound having a selectivity for ITK that is at least 50-fold greater than the selectivity for Bruton’s tyrosine kinase (BTK). In embodiments, the ITK inhibitor is a compound having a selectivity for ITK that is at least 60-fold greater than the selectivity for BTK. In embodiments, the ITK inhibitor is a compound having a selectivity for ITK that is at least 70-fold greater than the selectivity for BTK. In embodiments, the ITK inhibitor is a compound having a selectivity for ITK that is at least 80-fold greater than the selectivity for BTK. In embodiments, the ITK inhibitor is a compound having a selectivity for ITK that is at least 90-fold greater than the selectivity for BTK. In embodiments, the ITK inhibitor is a compound having a selectivity for ITK that is at least 100-fold greater than the selectivity for BTK.

[0097] In embodiments, the ITK inhibitor has a KD ≤ 10 nM to ITK. In embodiments, the ITK inhibitor has a KD ≤ 9 nM to ITK. In embodiments, the ITK inhibitor has a KD ≤ 8 nM to ITK. In embodiments, the ITK inhibitor has a KD ≤ 7 nM to ITK. In embodiments, the ITK inhibitor has a KD ≤ 6 nM to ITK. In embodiments, the ITK inhibitor has a KD ≤ 5 nM to ITK. In embodiments, the ITK inhibitor has a KD ≤ 4 nM to ITK. In embodiments, the ITK inhibitor has a KD ≤ 3 nM to ITK. In embodiments, the ITK inhibitor has a KD ≤ 2 nM to ITK. In embodiments, the ITK inhibitor has a KD ≤ 1 nM to ITK. In embodiments, the ITK inhibitor has a KD greater than 10 nM to receptor-like kinase (RLK). In embodiments, the ITK inhibitor has a KD greater than 100 nM to RLK. In embodiments, the ITK inhibitor has a KD greater than 1,000 nM to RLK. In embodiments, the ITK inhibitor has a KD greater than 10 nM to Bruton’s tyrosine kinase (BTK). In embodiments, the ITK inhibitor has a KD greater than 100 nM to BTK. In embodiments, the ITK inhibitor has a KD greater than 1,000 nM to BTK. In embodiments, the ITK inhibitor has any KD described herein for ITK and RLK. In embodiments, the ITK inhibitor has any KD described herein for ITK and BTK. In embodiments, the ITK inhibitor has any KD described herein for ITK, RLK, and BTK.

[0098] In embodiments, the ITK inhibitor has an IC50 ≤ 10 nM to ITK. In embodiments, the ITK inhibitor has an IC50 ≤ 9 nM to ITK. In embodiments, the ITK inhibitor has an IC50 ≤ 8 nM to ITK. In embodiments, the ITK inhibitor has an IC50 ≤ 7 nM to ITK. In embodiments, the ITK inhibitor has an IC50 ≤ 6 nM to ITK. In embodiments, the ITK inhibitor has an IC50 ≤ 5 nM to ITK. In embodiments, the ITK inhibitor has an IC50 ≤ 4 nM to ITK. In embodiments, the ITK inhibitor has an IC50 ≤ 3 nM to ITK. In embodiments, the ITK inhibitor has an IC50 ≤ 2 nM toITK. In embodiments, the ITK inhibitor has an IC50≤ 1 nM to ITK. In embodiments, the ITK inhibitor has an IC50 greater than 10 nM to RLK. In embodiments, the ITK inhibitor has an IC50 greater than 100 nM to RLK. In embodiments, the ITK inhibitor has an IC50greater than 1,000 nM to RLK. In embodiments, the ITK inhibitor has an IC50 greater than 10 nM to Bruton’s tyrosine kinase (BTK). In embodiments, the ITK inhibitor has an IC50greater than 100 nM to BTK. In embodiments, the ITK inhibitor has an IC50 greater than 1,000 nM to BTK. In embodiments, the ITK inhibitor has any IC50described herein for ITK and RLK. In embodiments, the ITK inhibitor has any IC50 described herein for ITK and BTK. In embodiments, the ITK inhibitor has any IC50described herein for ITK, RLK, and BTK. Methods for determining the KD or IC50 of kinase inhibitors (such as BTK, RLK, and ITK inhibitors) are well known in the art. See Fabian et al, Nat Biotechnol, 23:329-336 (2005)(doi:10.1038 / nbt1068) and Hsu et al, bioRxiv July 6, 2023 (doi.org / 10.1101 / 2023.07.05.547822). See also, e.g., Estupinan et al, Frontiers in Cell and Developmental Biology, Volume 9, Article 630942 (March 2021).

[0099] In embodiments, the ITK inhibitor is any compound described in US Patent No. 11,008,314, the disclosure of which is incorporated by reference herein in its entirety and for all purposes.

[0100] In embodiments, the ITK inhibitor is soquelitinib or a pharmaceutically acceptable salt thereof. In embodiments, the ITK inhibitor is the free base form of soquelitinib. In embodiments, the ITK inhibitor is a pharmaceutically acceptable salt of soquelitinib.

[0101] In embodiments, the ITK inhibitor is a compound of Formula (I) or a pharmaceutically acceptable salt thereof: ; wherein the substituents are

[0102] Ring A is, II) or a pharmaceutically acceptable salt thereof: ; wherein the substituents are as

[0104] R1, R2, R3, and R4are each independently hydrogen, halogen, -CX13, -CHX12, -CH2X1, -OCX13, -OCH2X1, -OCHX12, -CN, -SOn1R1A, -SOv1NR1AR1B, -NHC(O)NR1AR1B, -N(O)m1, -NR1AR1B, -C(O)R1A, -C(O)-OR1A, -C(O)NR1AR1B, -OR1A, -NR1ASO2R1B, -NR1AC(O)R1B, -NR1AC(O)OR1B, -NR1AOR1B, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

[0105] In embodiments, R1and R3are each independently hydrogen, halogen -CX13, -CHX12, -CH2X1, -OCX13, -OCH2X1, -OCHX12, -CN, -SOn1R1A, -SOv1NR1AR1B, -NHC(O)NR1AR1B, -N(O)m1, -NR1AR1B, -C(O)R1A, -C(O)-OR1A, -C(O)NR1AR1B, -OR1A, -NR1ASO2R1B, -NR1AC(O)R1B, -NR1AC(O)OR1B, or -NR1AOR1B.

[0106] In aspects, one or both of R1and R3are -OH. In aspects, one or both of R1and R3are -NH2. In aspects, one or both of R1and R3are -COOH. In aspects, one or both of R1and R3are -CONH2. In aspects, one or both of R1and R3are -NO2. In aspects, one or both of R1and R3are -SH. In aspects, one or both of R1and R3are -CF3. In aspects, one or both of R1and R3are -CHF2. In aspects, one or both of R1and R3are -CH2F. In aspects, one or both of R1and R3are -OCF3. In aspects, one or both of R1and R3are -OCH2F. In aspects, one or both of R1and R3are -OCHF2. In aspects, one or both of R1and R3are -OCH3. In aspects, one or both of R1and R3are -OCH2CH3. In aspects, one or both of R1and R3are -OCH2CH2CH3. In aspects, one or both of R1and R3are –OCH(CH3)2. In aspects, one or both of R1and R3are –OC(CH3)3. In aspects, one or both of R1and R3are –SCH3. In aspects, one or both of R1and R3are – SCH2CH3. In aspects, one or both of R1and R3are –SCH2CH2CH3. In aspects, one or both of R1and R3are –SCH(CH3)2. In aspects, one or both of R1and R3are –SC(CH3)3. In aspects, one or both of R1and R3are –CH3. In aspects, one or both of R1and R3are –CH2CH3. In aspects, one or both of R1and R3are –CH2CH2CH3. In aspects, one or both of R1and R3are –CH(CH3)2. In aspects, one or both of R1and R3are –C(CH3)3. In aspects, one or both of R1and R3are –F. In aspects, one or both of R1and R3are –Cl. In aspects, one or both of R1and R3are –Br. In aspects, one or both of R1and R3are –I. In aspects, one or both of R1and R3are hydrogen, methyl, ethyl, propyl, -CN, -COOH, -CONH2, -F, -Cl, -Br, or -I.

[0107] R5is independently substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

[0108] In aspects, R5is substituted or unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2), substituted or unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted or unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6), substituted or unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted or unsubstituted aryl (e.g., C6-C10 or phenyl), or substituted or unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).

[0109] In aspects, R5is substituted or unsubstituted (C1-C8) alkyl, substituted or unsubstituted 2 to 8 membered heteroalkyl, substituted or unsubstituted (C3-C6) cycloalkyl, substituted or unsubstituted 3 to 6 membered heterocycloalkyl, substituted or unsubstituted phenyl, or substituted or unsubstituted 5 to 6 membered heteroaryl. In aspects, R5is substituted or unsubstituted (C1-C4) alkyl. In aspects, R5is unsubstituted (C1-C4) alkyl. In aspects, R5is unsubstituted methyl, unsubstituted ethyl, unsubstituted isopropyl, or unsubstituted tert-butyl. Inaspects, R5is substituted or unsubstituted 2 to 8 membered heteroalkyl. In aspects, R5is substituted or unsubstituted 2 to 4 membered heteroalkyl. In aspects, R5is unsubstituted 2 to 4 membered heteroalkyl. In aspects, R5is -CH2N(CH3)2. In aspects, R5is substituted or unsubstituted (C3-C6) cycloalkyl. In aspects, R5is unsubstituted (C3-C6) cycloalkyl. In aspects, R5is unsubstituted cyclopropyl, unsubstituted cyclobutyl, or unsubstituted cyclopentyl. In aspects, R5is substituted or unsubstituted 3 to 6 membered heterocycloalkyl. In aspects, R5is substituted or unsubstituted 5 to 6 membered heterocycloalkyl. In aspects, R5is substituted or unsubstituted 6 membered heterocycloalkyl. In aspects, R5is substituted or unsubstituted piperidinyl. In aspects, R5is substituted or unsubstituted phenyl. In aspects, R5is unsubstituted phenyl. In aspects, R5is 2-substituted phenyl. In aspects, R5is 3-substituted phenyl. In aspects, R5is 4-substituted phenyl. In aspects, R5is phenyl substituted with halogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl. In aspects, R5is phenyl substituted with halogen, substituted or unsubstituted (C1-C8) alkyl, substituted or unsubstituted 2 to 8 membered heteroalkyl, substituted or unsubstituted (C3-C6) cycloalkyl, substituted or unsubstituted 3 to 6 membered heterocycloalkyl, substituted or unsubstituted phenyl, or substituted or unsubstituted 5 to 6 membered heteroaryl. In aspects, R5is –N(CH3)2. In aspects, R5is –NH(CH3). In aspects, R5is –N(CH2CH3)2. In aspects, R5is –NH(CH2CH3). In aspects, R5is –N(CH3)(CH2CH3). In aspects, R5is –CH3. In aspects, R5is –CH2CH3. In aspects, R5is unsubstituted isopropyl. In aspects, R5is unsubstituted tert-butyl.

[0110] In aspects, R5is substituted or unsubstituted heteroaryl. In aspects, R5is substituted or unsubstituted 5 to 6 membered heteroaryl. In aspects, R5is substituted or unsubstituted pyridyl, substituted or unsubstituted thienyl, substituted or unsubstituted furanyl, substituted or unsubstituted pyrrolyl, substituted or unsubstituted imidazolyl, substituted or unsubstituted pyrazolyl, substituted or unsubstituted thiazolyl, substituted or unsubstituted isothiazolyl, substituted or unsubstituted oxazolyl, or substituted or unsubstituted isoxazolyl.

[0111] In aspects, R5is substituted or unsubstituted cycloalkyl (e.g., C3-C8cycloalkyl, C3-C6cycloalkyl, or C5-C6 cycloalkyl). In aspects, R5is substituted or unsubstituted C3-C8 cycloalkyl. In aspects, R5is substituted or unsubstituted C3-C6cycloalkyl. In aspects, R5is substituted or unsubstituted C5-C6 cycloalkyl. In aspects, R5is substituted or unsubstituted C6 cycloalkyl. In aspects, R5is substituted or unsubstituted C5cycloalkyl. In aspects, R5is a substituted cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl). In aspects, R5is a substituted C3- C8cycloalkyl. In aspects, R5is a substituted C3-C6cycloalkyl. In aspects, R5is a substituted C5-C6cycloalkyl. In aspects, R5is a substituted C6cycloalkyl. In aspects, R5is a substituted C5cycloalkyl. In aspects, R5is an unsubstituted cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl, or C5-C6cycloalkyl). In aspects, R5is an unsubstituted C3-C8cycloalkyl. In aspects, R5is an unsubstituted C3-C6 cycloalkyl. In aspects, R5is an unsubstituted C5-C6 cycloalkyl. In aspects, R5is an unsubstituted C6cycloalkyl. In aspects, R5is an unsubstituted C5cycloalkyl.

[0112] In aspects, R5is substituted or unsubstituted aziridinyl, substituted or unsubstituted oziranyl, substituted or unsubstituted thiiranyl, substituted or unsubstituted azetidinyl, substituted or unsubstituted 1,2-dihydroazotyl, substituted or unsubstituted oxetanyl, substituted or unsubstituted 2H-oxetyl, substituted or unsubstituted thietanyl, substituted or unsubstituted 2H-thietyl, substituted or unsubstituted pyrrolidinyl, substituted or unsubstituted 2,5-dihydro- 1H-pyrrolyl, substituted or unsubstituted 4,5-dihydro-1H-imidazolyl, substituted or unsubstituted imidazolinyl, substituted or unsubstituted pyrazolinyl, substituted or unsubstituted tetrahydrofuranyl, substituted or unsubstituted thiolanyl, substituted or unsubstituted piperidinyl, substituted or unsubstituted piperazinyl, substituted or unsubstituted 2H-pyranyl, substituted or unsubstituted morpholinyl, substituted or unsubstituted 1,4-dioxanyl, tetrahydro-2H-pyranyl, substituted or unsubstituted thianyl, or substituted or unsubstituted dithianyl. In aspects, R5is a substituted aziridinyl, a substituted oziranyl, a substituted thiiranyl, a substituted azetidinyl, a substituted 1,2-dihydroazotyl, a substituted oxetanyl, a substituted 2H-oxetyl, a substituted thietanyl, a substituted 2H-thietyl, a substituted pyrrolidinyl, a substituted 2,5-dihydro-1H- pyrrolyl, a substituted 4,5-dihydro-1H-imidazolyl, a substituted imidazolinyl, a substituted pyrazolinyl, a substituted tetrahydrofuranyl, a substituted thiolanyl, a substituted piperidinyl, a substituted piperazinyl, a substituted 2H-pyranyl, a substituted morpholinyl, a substituted 1,4- dioxanyl, tetrahydro-2H-pyranyl, a substituted thianyl, or a substituted dithianyl. In aspects, R5is an unsubstituted aziridinyl, an unsubstituted oziranyl, an unsubstituted thiiranyl, an unsubstituted azetidinyl, an unsubstituted 1,2-dihydroazotyl, an unsubstituted oxetanyl, an unsubstituted 2H-oxetyl, an unsubstituted thietanyl, an unsubstituted 2H-thietyl, an unsubstituted pyrrolidinyl, an unsubstituted 2,5-dihydro-1H-pyrrolyl, an unsubstituted 4,5- dihydro-1H-imidazolyl, an unsubstituted imidazolinyl, an unsubstituted pyrazolinyl, an unsubstituted tetrahydrofuranyl, an unsubstituted thiolanyl, an unsubstituted piperidinyl, an unsubstituted piperazinyl, an unsubstituted 2H-pyranyl, an unsubstituted morpholinyl, an unsubstituted 1,4-dioxanyl, tetrahydro-2H-pyranyl, an unsubstituted thianyl, or an unsubstituted dithianyl.

[0113] In aspects, R5is substituted or unsubstituted (C6-C10) aryl. In aspects, R5is substituted or unsubstituted phenyl. In aspects, R5is substituted or unsubstituted naphthyl. In aspects, R5isa substituted (C6-C10) aryl. In aspects, R5is a substituted phenyl. In aspects, R5is a substituted naphthyl. In aspects, R5is an unsubstituted (C6-C10) aryl. In aspects, R5is an unsubstituted phenyl. In aspects, R5is an unsubstituted naphthyl.

[0114] In aspects, R5is imidazolyl, substituted or unsubstituted pyrrolyl, substituted or unsubstituted pyrazolyl, substituted or unsubstituted triazolyl, substituted or unsubstituted tetrazolyl, substituted or unsubstituted furanyl, substituted or unsubstituted oxazolyl, substituted or unsubstituted isooxazolyl, substituted or unsubstituted oxadiazolyl, substituted or unsubstituted oxatriazolyl, substituted or unsubstituted thienyl, substituted or unsubstituted thiazolyl, substituted or unsubstituted isothiazolyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted pyrazinyl, substituted or unsubstituted pyrimidinyl, substituted or unsubstituted pyridazinyl, or substituted or unsubstituted triazinyl (e.g., 1,3,5-triazinyl, 1,2,3- triazinyl, or 1,2,4-triazinyl). In aspects, R5is imidazolyl, a substituted pyrrolyl, a substituted pyrazolyl, a substituted triazolyl, a substituted tetrazolyl, a substituted furanyl, a substituted oxazolyl, a substituted isooxazolyl, a substituted oxadiazolyl, a substituted oxatriazolyl, a substituted thienyl, a substituted thiazolyl, a substituted isothiazolyl, a substituted pyridinyl, a substituted pyrazinyl, a substituted pyrimidinyl, a substituted pyridazinyl, or a substituted triazinyl (e.g., 1,3,5-triazinyl, 1,2,3-triazinyl, or 1,2,4-triazinyl). In aspects, R5is imidazolyl, an unsubstituted pyrrolyl, an unsubstituted pyrazolyl, an unsubstituted triazolyl, an unsubstituted tetrazolyl, an unsubstituted furanyl, an unsubstituted oxazolyl, an unsubstituted isooxazolyl, an unsubstituted oxadiazolyl, an unsubstituted oxatriazolyl, an unsubstituted thienyl, an unsubstituted thiazolyl, an unsubstituted isothiazolyl, an unsubstituted pyridinyl, an unsubstituted pyrazinyl, an unsubstituted pyrimidinyl, an unsubstituted pyridazinyl, or an unsubstituted triazinyl (e.g., 1,3,5-triazinyl, 1,2,3-triazinyl, or 1,2,4-triazinyl).

[0115] In aspects, R5is .(e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, R5is substituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, R5is unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, R5is unsubstituted methyl. In aspects, R5is unsubstituted ethyl. In aspects, R5is unsubstituted propyl. In aspects, R5is unsubstituted isopropyl. In aspects, R5is unsubstituted tert-butyl. In aspects, R5is substituted or unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, R5is substituted heteroalkyl (e.g., 2 to 8 membered,2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, R5is unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, R5is substituted or unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). In aspects, R5is substituted cycloalkyl (e.g., C3-C8, C3-C6, C4- C6, or C5-C6). In aspects, R5is unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). In aspects, R5is substituted or unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, R5is substituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, R5is unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, R5is substituted or unsubstituted aryl (e.g., C6-C10or phenyl). In aspects, R5is substituted aryl (e.g., C6-C10 or phenyl). In aspects, R5is unsubstituted aryl (e.g., C6-C10 or phenyl). In aspects, R5is substituted or unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, R5is substituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, R5is unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).

[0117] In aspects, R5is substituted or unsubstituted pyrrolidinyl. In aspects, R5is substituted or unsubstituted tetrahydrofuranyl. In aspects, R5is substituted or unsubstituted imidazolidinyl. In aspects, R5is substituted or unsubstituted pyrazolidinyl. In aspects, R5is substituted or unsubstituted oxazolidinyl. In aspects, R5is substituted or unsubstituted isoxazolidinyl. In aspects, R5is substituted or unsubstituted thiazolidinyl. In aspects, R5is substituted or unsubstituted isothiazolidinyl. In aspects, R5is substituted or unsubstituted dioxolanyl. In aspects, R5is substituted or unsubstituted dithiolanyl. In aspects, R5is substituted or unsubstituted piperidinyl. In aspects, R5is substituted or unsubstituted oxanyl. In aspects, R5is substituted or unsubstituted piperazinyl. In aspects, R5is substituted or unsubstituted morpholinyl. In aspects, R5is substituted or unsubstituted pyridinyl. In aspects, R5is substituted or unsubstituted triazolyl. In aspects, R5is substituted or unsubstituted tetrazolyl. In aspects, R5is substituted or unsubstituted benzo[d][1,3]dioxolyl. In aspects, R5is substituted or unsubstituted phenyl. In aspects, R5is substituted or unsubstituted pyridyl. In aspects, R5is substituted or unsubstituted pyridazinyl. In aspects, R5is substituted or unsubstituted pyrimidinyl. In aspects, R5is substituted or unsubstituted pyrazinyl. In aspects, R5is substituted or unsubstituted piperidinyl. In aspects, R5is substituted or unsubstituted tetrahydropyranyl. In aspects, R5is substituted or unsubstituted tetrahydrothiopyranyl. In aspects, R5is substituted or unsubstituted cyclohexyl. In aspects, R5is substituted or unsubstituted cyclopentyl. In aspects,R5is substituted or unsubstituted cycloheptyl. In aspects, R5is substituted or unsubstituted cyclobutyl. In aspects, R5is substituted or unsubstituted cyclopropyl. In aspects, R5is substituted or unsubstituted pyrrolyl. In aspects, R5is substituted or unsubstituted furanyl. In aspects, R5is substituted or unsubstituted thienyl. In aspects, R5is substituted or unsubstituted pyrazolyl. In aspects, R5is substituted or unsubstituted imidazolyl. In aspects, R5is substituted or unsubstituted isoxazolyl. In aspects, R5is substituted or unsubstituted oxazolyl. In aspects, R5is substituted or unsubstituted isothiazolyl. In aspects, R5is substituted or unsubstituted thiazolyl. In aspects, R5is substituted or unsubstituted naphthyl. In aspects, R5is substituted or unsubstituted quinolinyl. In aspects, R5is substituted or unsubstituted isoquinolinyl. In aspects, R5is substituted or unsubstituted indolyl. In aspects, R5is substituted or unsubstituted benzimidazolyl. In aspects, R5is substituted or unsubstituted indazolyl. In aspects, R5is substituted or unsubstituted isoindolyl. In aspects, R5is substituted or unsubstituted benzofuranyl. In aspects, R5is substituted or unsubstituted benzo[c]thienyl. In aspects, R5is substituted or unsubstituted 2,3-dihydro-1H-indenyl. In aspects, R5is substituted or unsubstituted 1,2,3,4-tetrahydronaphthyl. In aspects, R5is substituted or unsubstituted triazolyl. In aspects, R5is substituted or unsubstituted quinoxalinyl. In aspects, R5is substituted or unsubstituted quinazolinyl. In aspects, R5is substituted or unsubstituted triazinyl. In aspects, R5is substituted or unsubstituted cinnolinyl. In aspects, R5is substituted or unsubstituted phthalazinyl. In aspects, R5is substituted or unsubstituted benzoxazolyl. In aspects, R5is substituted or unsubstituted benzisoxazolyl. In aspects, R5is substituted or unsubstituted benzothiazolyl. In aspects, R5is substituted or unsubstituted benzisothiazolyl. In aspects, R5is substituted or unsubstituted benzo[d][1,2,3]triazolyl. In aspects, R5is substituted or unsubstituted adamantyl.

[0118] L1is -O-, -S-, or substituted or unsubstituted C1-C2alkylene, or substituted or unsubstituted 2 membered heteroalkylene. In aspect, L1is –O-. In aspect, L1is –S-. In aspect, L1is substituted C1-C2alkylene. In aspect, L1is unsubstituted C1-C2alkylene. In aspect, L1is substituted 2 membered heteroalkylene. In aspect, L1is unsubstituted 2 membered heteroalkylene.

[0119] L2is a bond, -NH-, -C(O)NH-, or -NHC(O)-. In aspects, L2is a bond. In aspects, L2is -NH-. In aspects, L2is -NHC(O)-.

[0120] L3and L4are each independently a bond, -S(O)2-, -N(R6)-, -O-, -S-, -C(O)-, -C(O)N(R6)-, -N(R6)C(O)-, -N(R6)C(O)NH-, -NHC(O)N(R6)-, -C(O)O-, -OC(O)-, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstitutedarylene, or substituted or unsubstituted heteroarylene.

[0121] In aspects, L3is a bond, -N(R6)-, -C(O)-, -C(O)N(R6)-, -N(R6)C(O)-, substituted or unsubstituted alkylene (e.g., C1-C8, C1-C6, C1-C4, or C1-C2), substituted or unsubstituted heteroalkylene (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted or unsubstituted cycloalkylene (e.g., C3-C8, C3-C6, C4-C6, or C5- C6), substituted or unsubstituted heterocycloalkylene (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted or unsubstituted arylene (e.g., C6-C10 or phenyl), or substituted or unsubstituted heteroarylene (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).

[0122] In aspects, L3is a bond, -N(R6)-, -C(O)-, or -C(O)N(R6)-; and R6is hydrogen, -CX63, -CHX62, -CH2X6, or unsubstituted (C1-C4) alkyl. In aspects, L3is a bond, -N(R6)-, -C(O)-, or -C(O)N(R6)-; and R6is hydrogen or unsubstituted methyl. In aspects, L3is a bond, -C(O)-, -C(O)N(CH3)-, -N(CH3)-, or -NH-. In aspects, L3is a bond. In aspects, L3is -C(O)-. In aspects, L3is -N(R6)-. In aspects, L3is -C(O)-. In aspects, L3is -C(O)N(R6)-. In aspects, L3is -NH-. In aspects, L3is -C(O)-. In aspects, L3is -C(O)NH-. In aspects, L3is -N(CH3)-. In aspects, L3is -C(O)N(CH3)-. In aspects, L3is -N(CH2CH3)-. In aspects, L3is -C(O)N(CH2CH3)-.

[0123] In aspects, L3is substituted or unsubstituted alkylene (e.g., C1-C8, C1-C6, C1-C4, or C1- C2). In aspects, L3is substituted alkylene (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, L3is unsubstituted alkylene (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, L3is unsubstituted methylene. In aspects, L3is unsubstituted ethylene. In aspects, L3is unsubstituted propylene. In aspects, L3is unsubstituted isopropylene. In aspects, L3is unsubstituted tert-butylene. In aspects, L3is substituted or unsubstituted heteroalkylene (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, L3is substituted heteroalkylene (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, L3is unsubstituted heteroalkylene (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, L3is substituted or unsubstituted cycloalkylene (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). In aspects, L3is substituted cycloalkylene (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). In aspects, L3is unsubstituted cycloalkylene (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). In aspects, L3is substituted or unsubstituted heterocycloalkylene (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, L3is substituted heterocycloalkylene (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, L3is unsubstituted heterocycloalkylene (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, L3is substituted or unsubstitutedarylene (e.g., C6-C10or phenylene). In aspects, L3is substituted arylene (e.g., C6-C10or phenylene). In aspects, L3is unsubstituted arylene (e.g., C6-C10 or phenylene). In aspects, L3is substituted or unsubstituted heteroarylene (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, L3is substituted heteroarylene (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, L3is unsubstituted heteroarylene (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).

[0124] In aspects, L3is a bond, -S(O)2-, -N(R6)-, -O-, -S-, -C(O)-, -C(O)N(R6)-, -N(R6)C(O)-, -N(R6)C(O)NH-, -NHC(O)N(R6)-, -C(O)O-, -OC(O)-, a substituted or unsubstituted alkylene (e.g., C1-C8, C1-C6, C1-C4, or C1-C2), a substituted or unsubstituted heteroalkylene (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), a substituted or unsubstituted cycloalkylene (e.g., C3-C8, C3-C6, C4-C6, or C5-C6), a substituted or unsubstituted heterocycloalkylene (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), a substituted or unsubstituted arylene (e.g., C6-C10 or phenylene), or a substituted or unsubstituted heteroarylene (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, L3is a bond, -S(O)2-, -N(R6)-, -O-, -S-, -C(O)-, -C(O)N(R6)-, -N(R6)C(O)-, -N(R6)C(O)NH-, -NHC(O)N(R6)-, -C(O)O-, -OC(O)-, unsubstituted alkylene (e.g., C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C10 or phenylene), or unsubstituted heteroarylene (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, L3is unsubstituted methylene. In aspects, L3is unsubstituted ethylene. In aspects, L3is methyl-substituted methylene.

[0125] In aspects, L4is a bond, -N(R6)-, -C(O)-, -C(O)N(R6)-, -N(R6)C(O)-, substituted or unsubstituted alkylene (e.g., C1-C8, C1-C6, C1-C4, or C1-C2), substituted or unsubstituted heteroalkylene (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted or unsubstituted cycloalkylene (e.g., C3-C8, C3-C6, C4-C6, or C5- C6), substituted or unsubstituted heterocycloalkylene (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted or unsubstituted arylene (e.g., C6-C10 or phenyl), or substituted or unsubstituted heteroarylene (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).

[0126] In aspects, L4is a bond. In aspects, L4is -N(R6)-. In aspects, L4is -C(O)N(R6)-. In aspects, L4is -NH-. In aspects, L4is -C(O)-. In aspects, L4is -C(O)NH-. In aspects, L4is -N(CH3)-. In aspects, L4is -C(O)N(CH3)-. In aspects, L4is -N(CH2CH3)-. In aspects, L4is -C(O)N(CH2CH3)-. In aspects, L4is a bond, -N(R7)-, -C(O)-, -C(O)N(R7)-, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted fused ring heterocycloalkylene, substituted or unsubstituted spirocyclic heterocycloalkylene, or substituted or unsubstituted bridged ring heterocycloalkylene; and R6is hydrogen, -CX63, -CHX62, -CH2X6, or unsubstituted (C1-C4) alkyl.

[0127] In aspects, L4is a bond, substituted or unsubstituted monocyclic heterocycloalkylene, substituted or unsubstituted fused ring heterocycloalkylene, substituted or unsubstituted spirocyclic heterocycloalkylene, or substituted or unsubstituted bridged ring heterocycloalkylene. In aspects, L4is unsubstituted 7 to 8 membered bridged ring heterocycloalkylene. In aspects, L4is unsubstituted 7 to 8 membered fused ring heterocycloalkylene. In aspects, L4is unsubstituted 7 to 8 membered spirocyclic heterocycloalkylene. In aspects, L4is unsubstituted 5 to 8 membered monocyclic heterocycloalkylene. In aspects, L4is methyl-substituted 5 to 8 membered monocyclic heterocycloalkylene. In aspects, L4is methyl-substituted 6 to 7 membered monocyclic heterocycloalkylene. In aspects, L4is ethyl-substituted 5 to 8 membered monocyclic heterocycloalkylene. In aspects, L4is ethyl-substituted 6 to 7 membered monocyclic heterocycloalkylene. In aspects, L4is cyano-substituted 5 to 8 membered monocyclic heterocycloalkylene. In aspects, L4is cyano-substituted 6 to 7 membered monocyclic heterocycloalkylene. In aspects, L4is halo-substituted 5 to 8 membered monocyclic heterocycloalkylene. In aspects, L4is halo-substituted 6 to 7 membered monocyclic heterocycloalkylene. In aspects, L4is unsubstituted 5 to 8 membered monocyclic heterocycloalkylene. In aspects, L4is unsubstituted 6 to 7 membered monocyclic heterocycloalkylene. In aspects, L4is ethyl-substituted 5 to 8 membered monocyclic heterocycloalkylene. In aspects, L4is ethyl-substituted 6 to 7 membered monocyclic heterocycloalkylene. In aspects, L4is isopropyl-substituted 5 to 8 membered monocyclic heterocycloalkylene. In aspects, L4is isopropyl-substituted 6 to 7 membered monocyclic heterocycloalkylene. In aspects, L4is tert-butyl-substituted 5 to 8 membered monocyclic heterocycloalkylene. In aspects, L4is a tert-butyl-substituted 6 to 7 membered monocyclic heterocycloalkylene. In aspects, L4is substituted or unsubstituted 4 to 10 membered monocyclic heterocycloalkylene. In aspects, L4is substituted or unsubstituted 5 to 8 membered monocyclic heterocycloalkylene. In aspects, L4is substituted 5 to 8 membered monocyclic heterocycloalkylene. In aspects, L4is unsubstituted 5 to 8 membered monocyclic heterocycloalkylene. In aspects, L4is substituted or unsubstituted 5 to 10 membered fused ringheterocycloalkylene. In aspects, L4is substituted or unsubstituted 7 to 8 membered fused ring heterocycloalkylene. In aspects, L4is substituted 5 to 10 membered fused ring heterocycloalkylene. In aspects, L4is substituted 7 to 8 membered fused ring heterocycloalkylene. In aspects, L4is unsubstituted 5 to 10 membered fused ring heterocycloalkylene. In aspects, L4is unsubstituted 7 to 8 membered fused ring heterocycloalkylene. In aspects, L4is unsubstituted 5 membered bridged ring heterocycloalkylene. In aspects, L4is unsubstituted 6 membered bridged ring heterocycloalkylene. In aspects, L4is unsubstituted 7 membered bridged ring heterocycloalkylene. In aspects, L4is unsubstituted 8 membered bridged ring heterocycloalkylene. In aspects, L4is unsubstituted 9 membered bridged ring heterocycloalkylene. In aspects, L4is unsubstituted 10 membered bridged ring heterocycloalkylene. In aspects, L4is unsubstituted 5 membered heterocycloalkylene. In aspects, L4is unsubstituted 6 membered heterocycloalkylene. In aspects, L4is unsubstituted 7 membered heterocycloalkylene. In aspects, L4is unsubstituted 8 membered heterocycloalkylene. In aspects, L4is unsubstituted 9 membered heterocycloalkylene. In aspects, L4is unsubstituted 10 membered heterocycloalkylene.

[0128] In aspects, L4is substituted or unsubstituted cycloalkylene. In aspects, L4is substituted or unsubstituted C3-C8 cycloalkylene. In aspects, L4is substituted or unsubstituted C3-C6 cycloalkylene. In aspects, L4is substituted or unsubstituted C5-C6 cycloalkylene. In aspects, L4is substituted or unsubstituted C6cycloalkylene. In aspects, L4is substituted or unsubstituted C5cycloalkylene. In aspects, L4is substituted cycloalkylene. In aspects, L4is substituted C3-C8 cycloalkylene. In aspects, L4is substituted C3-C6cycloalkylene. In aspects, L4is substituted C5- C6 cycloalkylene. In aspects, L4is substituted C6 cycloalkylene. In aspects, L4is substituted C5 cycloalkylene. In aspects, L4is an unsubstituted cycloalkylene. In aspects, L4is an unsubstituted C3-C8 cycloalkylene. In aspects, L4is an unsubstituted C3-C6 cycloalkylene. In aspects, L4is an unsubstituted C5-C6cycloalkylene. In aspects, L4is an unsubstituted C6cycloalkylene. In aspects, L4is an unsubstituted C5 cycloalkylene.

[0129] In aspects, L4is substituted or unsubstituted aziridinylene, substituted or unsubstituted oziranylene, substituted or unsubstituted thiiranylene, substituted or unsubstituted azetidinylene, substituted or unsubstituted 1,2-dihydroazotylene, substituted or unsubstituted oxetanylene, substituted or unsubstituted 2H-oxetylene, substituted or unsubstituted thietanylene, substituted or unsubstituted 2H-thietylene, substituted or unsubstituted pyrrolidinylene, substituted or unsubstituted 2,5-dihydro-1H-pyrrolylene, substituted or unsubstituted 4,5-dihydro-1H- imidazolylene, substituted or unsubstituted imidazolinylene, substituted or unsubstitutedpyrazolinylene, substituted or unsubstituted tetrahydrofuranylene, substituted or unsubstituted thiolanylene, substituted or unsubstituted piperidinylene, substituted or unsubstituted piperazinylene, substituted or unsubstituted 2H-pyranylene, substituted or unsubstituted morpholinylene, substituted or unsubstituted 1,4-dioxanylene, substituted or unsubstituted tetrahydro-2H-pyranylene, substituted or unsubstituted thianylene, or substituted or unsubstituted dithianylene. In aspects, L4is substituted aziridinylene, substituted oziranylene, substituted thiiranylene, substituted azetidinylene, substituted 1,2-dihydroazotylene, substituted oxetanylene, substituted 2H-oxetylene, substituted thietanylene, substituted 2H-thietylene, substituted pyrrolidinylene, substituted 2,5-dihydro-1H-pyrrolylene, substituted 4,5-dihydro-1H- imidazolylene, substituted imidazolinylene, substituted pyrazolinylene, substituted tetrahydrofuranylene, substituted thiolanylene, substituted piperidinylene, substituted piperazinylene, substituted 2H-pyranylene, substituted morpholinylene, substituted 1,4- dioxanylene, substituted tetrahydro-2H-pyranylene, substituted thianylene, or substituted dithianylene. In aspects, L4is an unsubstituted aziridinylene, an unsubstituted oziranylene, an unsubstituted thiiranylene, an unsubstituted azetidinylene, an unsubstituted 1,2- dihydroazotylene, an unsubstituted oxetanylene, an unsubstituted 2H-oxetylene, an unsubstituted thietanylene, an unsubstituted 2H-thietylene, an unsubstituted pyrrolidinylene, an unsubstituted 2,5-dihydro-1H-pyrrolylene, an unsubstituted 4,5-dihydro-1H-imidazolylene, an unsubstituted imidazolinylene, an unsubstituted pyrazolinylene, an unsubstituted tetrahydrofuranylene, an unsubstituted thiolanylene, an unsubstituted piperidinylene, an unsubstituted piperazinylene, an unsubstituted 2H-pyranylene, an unsubstituted morpholinylene, an unsubstituted 1,4- dioxanylene, an unsubstituted tetrahydro-2H-pyranylene, an unsubstituted thianylene, or an unsubstituted dithianylene.

[0130] In aspects, L4is substituted or unsubstituted (C6-C10) arylene. In aspects, L4is substituted or unsubstituted phenylene. In aspects, L4is substituted or unsubstituted naphthylene. In aspects, L4is substituted (C6-C10) arylene. In aspects, L4is substituted phenylene. In aspects, L4is substituted naphthylene. In aspects, L4is an unsubstituted (C6-C10) arylene. In aspects, L4is an unsubstituted phenylene. In aspects, L4is an unsubstituted naphthylene.

[0131]

[0132] In aspects, L4is substituted or unsubstituted imidazolylene, substituted or unsubstituted pyrrolylene, substituted or unsubstituted pyrazolylene, substituted or unsubstituted triazolylene, substituted or unsubstituted tetrazolylene, substituted or unsubstituted furanylene, substituted or unsubstituted oxazolylene, substituted or unsubstituted isooxazolylene, substituted or unsubstituted oxadiazolylene, substituted or unsubstituted oxatriazolylene, substituted orunsubstituted thienylene, substituted or unsubstituted thiazolylene, substituted or unsubstituted isothiazolylene, substituted or unsubstituted pyridinylene, substituted or unsubstituted pyrazinylene, substituted or unsubstituted pyrimidinylene, substituted or unsubstituted pyridazinylene, substituted or unsubstituted triazinylene (e.g., 1,3,5-triazinylene, 1,2,3- triazinylene, or 1,2,4-triazinylene). In aspects, L4is substituted imidazolylene, substituted pyrrolylene, substituted pyrazolylene, substituted triazolylene, substituted tetrazolylene, substituted furanylene, substituted oxazolylene, substituted isooxazolylene, substituted oxadiazolylene, substituted oxatriazolylene, substituted thienylene, substituted thiazolylene, substituted isothiazolylene, substituted pyridinylene, substituted pyrazinylene, substituted pyrimidinylene, substituted pyridazinylene, or substituted triazinylene (e.g., 1,3,5-triazinylene, 1,2,3-triazinylene, or 1,2,4-triazinylene). In aspects, L4is an unsubstituted imidazolylene, an unsubstituted pyrrolylene, an unsubstituted pyrazolylene, an unsubstituted triazolylene, an unsubstituted tetrazolylene, an unsubstituted furanylene, an unsubstituted oxazolylene, an unsubstituted isooxazolylene, an unsubstituted oxadiazolylene, an unsubstituted oxatriazolylene, an unsubstituted thienylene, an unsubstituted thiazolylene, an unsubstituted isothiazolylene, an unsubstituted pyridinylene, an unsubstituted pyrazinylene, an unsubstituted pyrimidinylene, an unsubstituted pyridazinylene, or an unsubstituted triazinylene (e.g., 1,3,5-triazinylene, 1,2,3- triazinylene, or 1,2,4-triazinylene).

[0133] In aspects, L4is isis. dependently hydrogen, -CX63, -CHX62, -CH2X6, -CN, -C(O)R6A, -C(O)OR6A, -C(O)NR6AR6B, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

[0135] In aspects, R6is independently hydrogen, -CX63, -CHX62, -CH2X6, -CN, -C(O)R6A, -C(O)-OR6A, -C(O)NR6AR6B, substituted or unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2), substituted or unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted or unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6), substituted or unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted or unsubstituted aryl (e.g., C6-C10or phenyl), or substituted or unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).

[0136] In aspects, R6is hydrogen. In aspects, R6is -CX63. In aspects, R6is -CHX62. In aspects, R6is -CH2X6. In aspects, R6is -CN. In aspects, R6is -C(O)R6A. In aspects, R6is -C(O)-OR6A. In aspects, R6is -C(O)NR6AR6B. In aspects, R6is -COOH. In aspects, R6is -CONH2. In aspects, R6is -CF3. In aspects, R6is -CHF2. In aspects, R6is -CH2F. In aspects, R6is –CH3. In aspects, R6is –CH2CH3. In aspects, R6is –CH2CH2CH3. In aspects, R6is –CH(CH3)2. In aspects, R6is – C(CH3)3.

[0137] In aspects, R6is substituted or unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, R6is substituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, R6is unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, R6is unsubstituted methyl. In aspects, R6is unsubstituted ethyl. In aspects, R6is unsubstituted propyl. In aspects, R6is unsubstituted isopropyl. In aspects, R6is unsubstituted tert-butyl. In aspects, R6is substituted or unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, R6is substituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, R6is unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, R6is substituted or unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). In aspects, R6is substituted cycloalkyl (e.g., C3-C8, C3-C6, C4- C6, or C5-C6). In aspects, R6is unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). Inaspects, R6is substituted or unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, R6is substituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, R6is unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, R6is substituted or unsubstituted aryl (e.g., C6-C10 or phenyl). In aspects, R6is substituted aryl (e.g., C6-C10or phenyl). In aspects, R6is unsubstituted aryl (e.g., C6-C10or phenyl). In aspects, R6is substituted or unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, R6is substituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, R6is unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).

[0138] E is an electrophilic moiety. In aspects, E is a covalent cysteine modifier moiety. In aspects, E is: In

[0139] R1A, R1B, R6A, and R6Bare each independently hydrogen, -CX3, -CN, -COOH, -CONH2, -CHX2, -CH2X, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R1Aand R1Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R6Aand R6Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl.

[0140] In aspects, R1A, R1B, R6A, and R6Bare each independently hydrogen, -CX1A3, -CHX1A2, -CH2X1A, -CN, -COOH, -CONH2, substituted or unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2), substituted or unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted or unsubstituted cycloalkyl(e.g., C3-C8, C3-C6, C4-C6, or C5-C6), substituted or unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted or unsubstituted aryl (e.g., C6-C10or phenyl), or substituted or unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, R1A, R1B, R6A, and R6Bare each independently hydrogen, -CX1A3, -CHX1A2, -CH2X1A, -CN, -COOH, -CONH2, unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted aryl (e.g., C6-C10 or phenyl), or unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, X1Ais independently –F, -Cl, -Br, or –I.

[0141] In aspects, one or more of R1A, R1B, R6A, and R6Bare hydrogen. In aspects, one or more of R1A, R1B, R6A, and R6Bare -CX63. In aspects, o one or more of R1A, R1B, R6A, and R6Bare - CHX62. In aspects, one or more of R1A, R1B, R6A, and R6Bare -CH2X6. In aspects, one or more of R1A, R1B, R6A, and R6Bare -CN. In aspects, one or more of R1A, R1B, R6A, and R6Bare -COOH. In aspects, one or more of R1A, R1B, R6A, and R6Bare -CONH2.

[0142] In aspects, one or more of R1A, R1B, R6A, and R6Bare substituted or unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, one or more of R1A, R1B, R6A, and R6Bare substituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, one or more of R1A, R1B, R6A, and R6Bare unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, one or more of R1A, R1B, R6A, and R6Bare unsubstituted methyl. In aspects, one or more of R1A, R1B, R6A, and R6Bare unsubstituted ethyl. In aspects, one or more of R1A, R1B, R6A, and R6Bare unsubstituted propyl. In aspects, one or more of R1A, R1B, R6A, and R6Bare unsubstituted isopropyl. In aspects, one or more of R1A, R1B, R6A, and R6Bare unsubstituted tert-butyl. In aspects, one or more of R1A, R1B, R6A, and R6Bare substituted or unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, one or more of R1A, R1B, R6A, and R6Bare substituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, one or more of R1A, R1B, R6A, and R6Bare unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, one or more of R1A, R1B, R6A, and R6Bare substituted or unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). In aspects, one or more of R1A, R1B, R6A, and R6Bare substituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). In aspects, one or more of R1A, R1B, R6A, and R6Bare unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). In aspects, one or more of R1A, R1B, R6A, and R6Bare substituted or unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, one or more of R1A, R1B, R6A, and R6Bare substituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, one or more of R1A, R1B, R6A, and R6Bare unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, one or more of R1A, R1B, R6A, and R6Bare substituted or unsubstituted aryl (e.g., C6-C10 or phenyl). In aspects, one or more of R1A, R1B, R6A, and R6Bare substituted aryl (e.g., C6-C10or phenyl). In aspects, one or more of R1A, R1B, R6A, and R6Bare unsubstituted aryl (e.g., C6-C10 or phenyl). In aspects, one or more of R1A, R1B, R6A, and R6Bare substituted or unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, one or more of R1A, R1B, R6A, and R6Bare substituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, o one or more of R1A, R1B, R6A, and R6Bare unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).

[0143] In aspects, one or more of R1A, R1B, R6A, and R6Bare hydrogen. In aspects, one or more of R1A, R1B, R6A, and R6Bare methyl. In aspects, one or more of R1A, R1B, R6A, and R6Bare ethyl. In aspects, one or more of R1A, R1B, R6A, and R6Bare propyl. In aspects, one or more of R1A, R1B, R6A, and R6Bare -CN. In as one or more of R1A, R1B, R6A, and R6Bare -CONH2. In aspects, one or more of R1A, R1B, R6A, and R6Bare –F, -Cl, -Br, or -I.

[0144] X, X1, X1A, X6, and X15are each independently –F, -Cl, -Br, or –I. In aspects, X is –F. In aspects, X is –Cl. In aspects, X is –Br. In aspects, X is –I. In aspects, X1is –F. In aspects, X1is –Cl. In aspects, X1is –Br. In aspects, X1is –I. In aspects, X1Ais –F. In aspects, X1Ais –Cl. In aspects, X1Ais –Br. In aspects, X1Ais –I. In aspects, X6is –F. In aspects, X6is –Cl. In aspects, X6is –Br. In aspects, X6is –I. In aspects, X15is –F. In aspects, X15is –Cl. In aspects, X15is –Br. In aspects, X15is –I.

[0145] n1 is each independently an integer from 0 to 4. In aspects, n1 is 0. In aspects, n1 is 1. In aspects, n1 is 2. In aspects, n1 is 3. In aspects, n1 is 4.

[0146] m1 is independently 1 or 2. In aspects, m1 is 1. In aspects, m1 is 2.

[0147] v1 is independently 1 or 2. In aspects, v1 is 1. In aspects, v1 is 2.

[0148] R15, R16, and R17are each independently hydrogen, halogen, -CX153, -CHX152, -CH2X15, -CN, -SOn15R15A, -SOv15NR15AR15B, ^NHNR15AR15B, -ONR15AR15B, -NHC=(O)NHNR15AR15B,^NHC(O)NR15AR15B, -N(O)m15, -NR15AR15B, -C(O)R15A,-C(O)-OR15A, -C(O)NR15AR15B, -OR15A, -NR15ASO2R15B, -NR15AC(O)R15B, -NR15AOR15B, -R15AC(O)OR15B, -OCX153, -OCHX152, -OCH2X15, substituted or unsubstituted alkyl (e.g., C1- C8, C1-C6, C1-C4, or C1-C2), substituted or unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted or unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6), substituted or unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted or unsubstituted aryl (e.g., C6-C10or phenyl), or substituted or unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).

[0149] In aspects, one or more of R15, R16, and R17are hydrogen. In aspects, one or more of R15, R16, and R17are halogen. In aspects, one or more of R15, R16, and R17are -CX153. In aspects, one or more of R15, R16, and R17are -CHX152. In aspects, one or more of R15, R16, and R17are - CH2X15. In aspects, one or more of R15, R16, and R17are –CN. In aspects, one or more of R15, R16, and R17are -SOn15R15A. In aspects, one or more of R15, R16, and R17are -SOv15NR15AR15B. In aspects, one or more of R15, R16, and R17are ^NHNR15AR15B. In aspects, one or more of R15, R16, and R17are ^ONR15AR15B. In aspects, one or more of R15, R16, and R17are ^NHC=(O)NHNR15AR15B. In aspects, one or more of R15, R16, and R17are^^NHC(O)NR15AR15B. In aspects, one or more of R15, R16, and R17are -N(O)m15. In aspects, one or more of R15, R16, and R17are -NR15AR15B. In aspects, one or more of R15, R16, and R17are -C(O)R15A. In aspects, one or more of R15, R16, and R17are -C(O)-OR15A. In aspects, one or more of R15, R16, and R17are -C(O)NR15AR15B. In aspects, one or more of R15, R16, and R17are -OR15A. In aspects, one or more of R15, R16, and R17are -NR15ASO2R15B. In aspects, one or more of R15, R16, and R17are - NR15AC(O)R15B. In aspects, one or more of R15, R16, and R17are -NR15AC(O)OR15B. In aspects, one or more of R15, R16, and R17are -NR15AOR15B. In aspects, one or more of R15, R16, and R17are -OCX153. In aspects, one or more of R15, R16, and R17are -OCHX152. In aspects, one or more of R15, R16, and R17are -OCH2X15. In aspects, one or more of R15, R16, and R17are -OH. In aspects, one or more of R15, R16, and R17are -NH2. In aspects, one or more of R15, R16, and R17are -COOH. In aspects, one or more of R15, R16, and R17are -CONH2. In aspects, one or more of R15, R16, and R17are -NO2. In aspects, one or more of R15, R16, and R17are -SH. In aspects, one or more of R15, R16, and R17are -CF3. In aspects, one or more of R15, R16, and R17are -CHF2. In aspects, one or more of R15, R16, and R17are -CH2F. In aspects, one or more of R15, R16, and R17are -OCF3. In aspects, one or more of R15, R16, and R17are -OCH2F. In aspects, one or more of R15, R16, and R17are -OCHF2. In aspects, one or more of R15, R16, and R17are –OCH3. In aspects, one or more of R15, R16, and R17are –OCH2CH3. In aspects, one or more of R15, R16, and R17are –OCH2CH2CH3. In aspects, one or more of R15, R16, and R17are –OCH(CH3)2. Inaspects, one or more of R15, R16, and R17are –OC(CH3)3. In aspects, one or more of R15, R16, and R17are –SCH3. In aspects, one or more of R15, R16, and R17are –SCH2CH3. In aspects, one or more of R15, R16, and R17are –SCH2CH2CH3. In aspects, one or more of R15, R16, and R17are –SCH(CH3)2. In aspects, one or more of R15, R16, and R17are –SC(CH3)3. In aspects, one or more of R15, R16, and R17are –CH3. In aspects, one or more of R15, R16, and R17are –CH2CH3. In aspects, one or more of R15, R16, and R17are –CH2CH2CH3. In aspects, one or more of R15, R16, and R17are –CH(CH3)2. In aspects, one or more of R15, R16, and R17are –C(CH3)3. In aspects, one or more of R15, R16, and R17are –F. In aspects, one or more of R15, R16, and R17are –Cl. In aspects, one or more of R15, R16, and R17are –Br. In aspects, one or more of R15, R16, and R17are –I.

[0150] In aspects, one or more of R15, R16, and R17are substituted or unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, one or more of R15, R16, and R17are substituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, one or more of R15, R16, and R17are unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, one or more of R15, R16, and R17are unsubstituted methyl. In aspects, one or more of R15, R16, and R17are unsubstituted ethyl. In aspects, one or more of R15, R16, and R17are unsubstituted propyl. In aspects, one or more of R15, R16, and R17are unsubstituted isopropyl. In aspects, one or more of R15, R16, and R17are unsubstituted tert-butyl. In aspects, one or more of R15, R16, and R17are substituted or unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, one or more of R15, R16, and R17are substituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, one or more of R15, R16, and R17are unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, one or more of R15, R16, and R17are substituted or unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). In aspects, one or more of R15, R16, and R17are substituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). In aspects, R15is unsubstituted cycloalkyl (e.g., C3-C8, C3- C6, C4-C6, or C5-C6). In aspects, one or more of R15, R16, and R17are substituted or unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, one or more of R15, R16, and R17are substituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, one or more of R15, R16, and R17are unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, one or more of R15, R16, and R17are substituted or unsubstituted aryl (e.g., C6-C10or phenyl). In aspects, one or more of R15, R16, and R17are substituted aryl(e.g., C6-C10or phenyl). In aspects, one or more of R15, R16, and R17are unsubstituted aryl (e.g., C6-C10 or phenyl). In aspects, one or more of R15, R16, and R17are substituted or unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, one or more of R15, R16, and R17are substituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, one or more of R15, R16, and R17are unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).

[0151] In aspects, R15, R16, and R17are each independently hydrogen, halogen, -CX153, -CHX152, -CH2X15, -OCX153, -OCH2X15, -OCHX152, -CN, -OH, -NH2, -COOH, -CONH2, -NO2, -SH, -SO3H, -SO4H, -SO2NH2, ^NHNH2, ^ONH2, ^NHC=(O)NHNH2, ^NHC=(O)NH2, -NHSO2H, -NHC=(O)H, -NHC(O)-OH, -NHOH, R15A-substituted or unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2), In aspects, R15, R16, and R17are each independently hydrogen, halogen, -CX153, -CHX152, -CH2X15, -OCX153, -OCH2X15, -OCHX152, -CN, -OH, -NH2, -COOH, -CONH2, -NO2, -SH, -SO3H, -SO4H, -SO2NH2, ^NHNH2, ^ONH2, -NHC(O)-OH, ^NHC=(O)NHNH2, ^NHC=(O) NH2, -NHSO2H, -NHC= (O)H, -NHOH, unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted aryl (e.g., C6-C10 or phenyl), or unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, one or more of R15, R16, and R17are hydrogen. In aspects, one or more of R15, R16, and R17are unsubstituted methyl. In aspects, one or more of R15, R16, and R17are unsubstituted ethyl.

[0152] In aspects, R15, R16, and R17are hydrogen. In aspects, R15is hydrogen. In aspects, R16is hydrogen, -CH3, or ^CH2NR15AR15B. In aspects, R17is hydrogen. In aspects, R15Aand R15Bare independently hydrogen or unsubstituted alkyl. In aspects, R15Aand R15Bare independently unsubstituted methyl. In aspects, R15Ais hydrogen. In aspects, R15Ais unsubstituted alkyl. In aspects, R15Bis hydrogen. In aspects, R15Bis unsubstituted alkyl. In aspects, R15Ais unsubstituted methyl. In aspects, R15Bis unsubstituted methyl. In aspects, R15is hydrogen; R16is hydrogen, -CH3, or ^CH2NR15AR15B; R17is hydrogen; and R15Aand R15Bare independently hydrogen or unsubstituted alkyl.

[0153] In aspects, R15is hydrogen. In aspects, R16is hydrogen. In aspects, R17is hydrogen, -CH3, or ^CH2NR15AR15B. In aspects, R15Aand R15Bare independently hydrogen or unsubstituted alkyl. In aspects, R15Aand R15Bare independently unsubstituted methyl. In aspects,R15is hydrogen; R16is hydrogen; R17is hydrogen, -CH3, or ^CH2NR15AR15B, and R15Aand R15Bare independently hydrogen or unsubstituted alkyl. In aspects, R15Ais hydrogen. In aspects, R15Ais unsubstituted alkyl. In aspects, R15Bis hydrogen. In aspects, R15Bis unsubstituted alkyl. In aspects, R15Ais unsubstituted methyl. In aspects, R15Bis unsubstituted methyl.

[0154] In aspects, R15is hydrogen, -CH3, or ^CH2NR15AR15B. In aspects, R16is hydrogen. In aspects, R17is hydrogen. In aspects, R15Aand R15Bare independently hydrogen or unsubstituted alkyl. In aspects, R15Aand R15Bare independently unsubstituted methyl. In aspects, R15Ais hydrogen. In aspects, R15Ais unsubstituted alkyl. In aspects, R15Bis hydrogen. In aspects, R15Bis unsubstituted alkyl. In aspects, R15Ais unsubstituted methyl. In aspects, R15Bis unsubstituted methyl. In aspects, R15is hydrogen, -CH3, or ^CH2NR15AR15B; R16is hydrogen; R17is hydrogen; and R15Aand R15Bare independently hydrogen or unsubstituted alkyl.

[0155] R15Aand R15Bare each independently hydrogen, -CX3, -CN, -COOH, -CONH2, -CHX2, -CH2X, substituted or unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2), substituted or unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted or unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4- C6, or C5-C6), substituted or unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted or unsubstituted aryl (e.g., C6-C10or phenyl), or substituted or unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered); R15Aand R15Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered) or substituted or unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).

[0156] In aspects, one or both of R15Aand R15Bare hydrogen. In aspects, one or both of R15Aand R15Bare -CX15A3. In aspects, one or both of R15Aand R15Bare -CHX15A2. In aspects, one or both of R15Aand R15Bare -CH2X15A. In aspects, one or both of R15Aand R15Bare -CN. In aspects, one or both of R15Aand R15Bare -COOH. In aspects, one or both of R15Aand R15Bare -CONH2.

[0157] In aspects, one or both of R15Aand R15Bare substituted or unsubstituted alkyl (e.g., C1- C8, C1-C6, C1-C4, or C1-C2). In aspects, one or both of R15Aand R15Bare substituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, one or both of R15Aand R15Bare unsubstituted alkyl (e.g., C1-C8, C1-C6, C1-C4, or C1-C2). In aspects, one or both of R15Aand R15Bare unsubstituted methyl. In aspects, one or both of R15Aand R15Bare unsubstituted ethyl. In aspects, one or bothof R15Aand R15Bare unsubstituted propyl. In aspects, one or both of R15Aand R15Bare unsubstituted isopropyl. In aspects, one or both of R15Aand R15Bare unsubstituted tert-butyl. In aspects, one or both of R15Aand R15Bare substituted or unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, one or both of R15Aand R15Bare substituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, one or both of R15Aand R15Bare unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In aspects, one or both of R15Aand R15Bare substituted or unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). In aspects, one or both of R15Aand R15Bare substituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). In aspects, one or both of R15Aand R15Bare unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6). In aspects, one or both of R15Aand R15Bare substituted or unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, one or both of R15Aand R15Bare substituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, one or both of R15Aand R15Bare unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered). In aspects, one or both of R15Aand R15Bare substituted or unsubstituted aryl (e.g., C6-C10 or phenyl). In aspects, one or both of R15Aand R15Bare substituted aryl (e.g., C6-C10or phenyl). In aspects, one or both of R15Aand R15Bare unsubstituted aryl (e.g., C6-C10or phenyl). In aspects, one or both of R15Aand R15Bare substituted or unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, one or both of R15Aand R15Bare substituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, one or both of R15Aand R15Bare unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).

[0158] In aspects, R15Aand R15Bare each independently hydrogen, -CX15A3, -CHX15A2, -CH2X15A, -CN, -COOH, -CONH2, In aspects, R15Aand R15Bare each independently hydrogen, -CX15A3, -CHX15A2, -CH2X15A, -CN, -COOH, -CONH2, unsubstituted alkyl (e.g., C1- C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkyl (e.g., 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkyl (e.g., C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkyl (e.g., 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted aryl (e.g., C6-C10or phenyl), or unsubstituted heteroaryl (e.g., 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In aspects, X15Ais independently –F, -Cl, -Br, or –I. In aspects, one or both of R15Aand R15Bare hydrogen. In aspects, one or both of R15Aand R15Bare unsubstituted methyl.In aspects, one or both of R15Aand R15Bare unsubstituted ethyl.

[0159] n15 is independently an integer from 0 to 4. In aspects, n15 is 0. In aspects, n15 is 1. In aspects, n15 is 2. In aspects, n15 is 3. In aspects, n15 is 4.

[0160] v15 is 1 or 2. In aspects, v15 is 1. In aspects, v15 is 2.

[0161] m15 is 1 or 2. In aspects m15 is 1. In aspects m15 is 2.

[0162] In embodiments, the compound of Formula (I) is soquelitinib.

[0163] In embodiments, the compound of Formula (I) is a compound of any one of Formula (1) to Formula (34) or a pharmaceutically acceptable salt of any one of the foregoing: , , ,,O S ,Patent No. 11,008,314, the disclosure of which is incorporated by reference herein in its entirety.

[0165] In embodiments, the ITK inhibitor is not ibrutinib. In embodiments, the ITK inhibitor is not BMS-509744 (CAS No.439575-02-7). In embodiments, the ITK inhibitor is not PRN694 (CAS No.1575818-46-0). In embodiments, the ITK inhibitor is not J-13 (the structure of which is shown in Figure 5 of Hantani et al, SLAS Discovery, 24(8):854-862 (2019)). In embodiments, the ITK inhibitor is not ECPIRM. In embodiments, the ITK inhibitor is not ibrutinib, BMS- 509744, PRN694, J-13, or ECPIRM. In embodiments, the ITK inhibitor is not an ITK inhibitor shown in Figure 3A of Hantani et al, SLAS Discovery, 24(8):854-862 (2019)).

[0166] In embodiments, the ITK inhibitor is ibrutinib. In embodiments, the ITK inhibitor is BMS-509744 (CAS No.439575-02-7). In embodiments, the ITK inhibitor is PRN694 (CAS No. 1575818-46-0). In embodiments, the ITK inhibitor is J-13 (the structure of which is shown in Figure 5 of Hantani et al, SLAS Discovery, 24(8):854-862 (2019)). In embodiments, the ITK inhibitor is ECPIRM. In embodiments, the ITK inhibitor is an ITK inhibitor shown in Figure 3A of Hantani et al, SLAS Discovery, 24(8):854-862 (2019)).

[0167] CAR T-Cells

[0168] CAR T-cells are known in the art and any CAR T-cells can be used in the methods, compositions, kits, and embodiments described herein.

[0169] In embodiments of the methods, compositions, and kits described herein, the CAR T- cells are a commercially available CAR T-cell therapy. In embodiments, the CAR T-cells are a CAR T-cell therapy selected from the group consisting of axicabtagene ciloleucel,brexucabtagene autoleucel, ciltacabtagene autoleucel, idecabtagene vicleucel, lisocabtagenemaraleucel, and tisagenlecleucel. In embodiments, the CAR T-cell therapy is axicabtageneciloleucel. In embodiments, the CAR T-cell therapy is brexucabtagene autoleucel. In embodiments, the CAR T-cell therapy is ciltacabtagene autoleucel. In embodiments, the CAR T- cell therapy is idecabtagene vicleucel. In embodiments, the CAR T-cell therapy is lisocabtagene maraleucel. In embodiments, the CAR T-cell therapy is tisagenlecleucel.

[0170] Chimeric antigen receptors (CAR) comprise an extracellular and intracellular domain. The extracellular domain comprises a target-specific binding element referred to as an antigen- binding moiety. The intracellular domain or otherwise the cytoplasmic domain comprises a costimulatory signaling region and a zeta chain portion. The costimulatory signaling region refers to a portion of the CAR comprising the intracellular domain of a costimulatory molecule. Costimulatory molecules are cell surface molecules other than antigens receptors or their ligands that are required for an efficient response of lymphocytes to antigen. Between the extracellular domain and the transmembrane domain of the CAR, or between the cytoplasmic domain and the transmembrane domain of the CAR, there may be incorporated a spacer domain. The term “spacer domain” generally means any oligo- or polypeptide that functions to link domains (e.g., the transmembrane domain to either the extracellular domain or the cytoplasmic domain in the polypeptide chain). A spacer domain may comprise up to 300 amino acids.

[0171] In embodiments, the binding region comprises complementary-determining regions (CDRs) of a monoclonal antibody, variable regions of a monoclonal antibody, and / or antigen- binding fragments thereof. A CDR is a short amino acid sequence found in the variable domains of antigen receptor (e.g., immunoglobulin and T-cell receptor) proteins that complements an antigen and therefore provides the receptor with its specificity for that particular antigen.

[0172] A CAR is engineered to target a tumor antigen of interest by way of engineering a desired antigen-binding moiety that specifically binds to an antigen on a tumor cell. As used herein, an “antigen” or “tumor antigen” or “cancer antigen,” refers to antigens that are common to specific hyperproliferative disorders such as cancer. Tumor antigens are moieties (e.g.,proteins, carbohydrates, etc.) that are produced by tumor cells that elicit an immune response, particularly T-cell mediated immune responses. Thus, an antigen-binding moiety can be selected based on the particular type of cancer to be treated. Tumor antigens are well known in the art. The tumor antigen may also be a tumor-specific antigen (TSA) or a tumor-associated antigen (TAA). A TSA is unique to tumor cells and does not occur on other cells in the body. A TAA is not unique to a tumor cell and instead is also expressed on some normal cells under conditions that fail to induce a state of immunologic tolerance to the antigen. The expression of the antigen on the tumor may occur under conditions that enable the immune system to respond to the antigen. TAAs may be antigens that are expressed on normal cells during fetal development when the immune system is immature and unable to respond or they may be antigens that are normally present at extremely low levels on normal cells but which are expressed at much higher levels on tumor cells.

[0173] In embodiments, the antigen binding domain binds to an antigen selected from the group consisting of CD19; CD123; CD22; CD30; CD171; CS-1; C-type lectin-like molecule-1, CD33; epidermal growth factor receptor variant III (EGFRvIII); ganglioside G2 (GD2); ganglioside GD3; TNF receptor family member; B-cell maturation antigen (BCMA); Tn antigen ((Tn Ag) or (GalNAcα-Ser / Thr)); prostate-specific membrane antigen (PSMA); Receptor tyrosine kinase-like orphan receptor 1 (ROR1); Fms-Like Tyrosine Kinase 3 (FLT3); Tumor- associated glycoprotein 72 (TAG72); CD38; CD44v6; Carcinoembryonic antigen (CEA); Epithelial cell adhesion molecule (EPCAM); B7H3 (CD276); KIT (CD117); Interleukin-13 receptor subunit alpha-2; mesothelin; Interleukin 11 receptor alpha (IL-11Ra); prostate stem cell antigen (PSCA); Protease Serine 21; vascular endothelial growth factor receptor 2 (VEGFR2); Lewis(Y) antigen; CD24; Platelet-derived growth factor receptor beta (PDGFR-beta); Stage- specific embryonic antigen-4 (SSEA-4); CD20; Folate receptor alpha; Receptor tyrosine-protein kinase ERBB2 (Her2 / neu); Mucin 1, cell surface associated (MUC1); epidermal growth factor receptor (EGFR); neural cell adhesion molecule (NCAM); Prostase; prostatic acid phosphatase (PAP); elongation factor 2 mutated (ELF2M); Ephrin B2; fibroblast activation protein alpha (FAP); insulin-like growth factor 1 receptor (IGF-I receptor), carbonic anhydrase IX (CAIX); Proteasome (Prosome, Macropain) Subunit, Beta Type, 9 (LMP2); glycoprotein 100 (gp100); oncogene fusion protein consisting of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcr-abl); tyrosinase; ephrin type-A receptor 2 (EphA2); Fucosyl GM1; sialyl Lewis adhesion molecule (sLe); ganglioside GM3; transglutaminase 5 (TGS5); high molecular weight-melanoma-associated antigen (HMWMAA); o-acetyl-GD2 ganglioside (OAcGD2); Folate receptor beta; tumor endothelial marker 1(TEM1 / CD248); tumor endothelial marker 7-related (TEM7R); claudin 6 (CLDN6); thyroid stimulating hormone receptor (TSHR); G protein-coupled receptor class C group 5, member D (GPRC5D); chromosome X open reading frame 61 (CXORF61); CD97; CD179a; anaplastic lymphoma kinase (ALK); Polysialic acid; placenta-specific 1 (PLAC1); hexasaccharide portion of globoH glycoceramide (GloboH); mammary gland differentiation antigen (NY-BR-1); uroplakin 2 (UPK2); Hepatitis A virus cellular receptor 1 (HAVCR1); adrenoceptor beta 3 (ADRB3); pannexin 3 (PANX3); G protein-coupled receptor 20 (GPR20); lymphocyte antigen 6 complex, locus K 9 (LY6K); Olfactory receptor 51E2 (OR51E2); TCR Gamma Alternate Reading Frame Protein (TARP); Wilms tumor protein (WT1); Cancer / testis antigen 1 (NY- ESO-1); Cancer / testis antigen 2 (LAGE-1a); Melanoma-associated antigen 1 (MAGE-A1); ETS translocation-variant gene 6, located on chromosome 12p (ETV6-AML); sperm protein 17 (SPA17); X Antigen Family, Member 1A (XAGE1); angiopoietin-binding cell surface receptor 2 (Tie 2); melanoma cancer testis antigen-1 (MAD-CT-1); melanoma cancer testis antigen-2 (MAD-CT-2); Fos-related antigen 1; tumor protein p53 (p53); p53 mutant; prostein; surviving; telomerase; prostate carcinoma tumor antigen-1, melanoma antigen recognized by T cells 1; Rat sarcoma (Ras) mutant; human Telomerase reverse transcriptase (hTERT); sarcoma translocation breakpoints; melanoma inhibitor of apoptosis (ML-IAP); ERG (transmembrane protease, serine 2 (TMPRSS2) ETS fusion gene); N-Acetyl glucosaminyl-transferase V (NA17); paired box protein Pax-3 (PAX3); Androgen receptor; Cyclin B1; v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog (MYCN); Ras Homolog Family Member C (RhoC); Tyrosinase-related protein 2 (TRP-2); Cytochrome P4501B1 (CYP1B1); CCCTC-Binding Factor (Zinc Finger Protein)-Like, Squamous Cell Carcinoma Antigen Recognized By T Cells 3 (SART3); Paired box protein Pax-5 (PAX5); proacrosin binding protein sp32 (OY-TES1); lymphocyte-specific protein tyrosine kinase (LCK); A kinase anchor protein 4 (AKAP-4); synovial sarcoma, X breakpoint 2 (SSX2); Receptor for Advanced Glycation Endproducts (RAGE-1); renal ubiquitous 1 (RU1); renal ubiquitous 2 (RU2); legumain; human papilloma virus E6 (HPV E6); human papilloma virus E7 (HPV E7); intestinal carboxyl esterase; heat shock protein 70-2 mutated (mut hsp70- 2); CD79a; CD79b; CD72; Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1); Fc fragment of IgA receptor (FCAR or CD89); Leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2); CD300 molecule- like family member f (CD300LF); C-type lectin domain family 12 member A (CLEC12A); bone marrow stromal cell antigen 2 (BST2); EGF-like module-containing mucin-like hormone receptor-like 2 (EMR2); lymphocyte antigen 75 (LY75); Glypican-3 (GPC3); Fc receptor-like 5 (FCRL5); immunoglobulin lambda-like polypeptide 1 (IGLL1); and a combination of two ormore thereof.

[0174] In embodiments, the antigen binding domain binds to an antigen selected from the group consistin of epidermal growth factor receptor, mesothelin, prostate-specific membrane antigen, disialoganglioside GD2, interleukin-13Ra2, glypican-3, carbonic anhydrase IX, L1CAM, fibroblast activation protein, a cancer testis antigen, MUC1, FOLR1, BCMA, CD133,alpha-fetoprotein, carcinoembryonic antigen, trophoblast glycoprotein precursor, immaturelaminin receptor protein, fetal sulphoglycoprotein, g-fetoprotein, cc2-H-ferroportin, beta-2- microglobulin, beta-human chorionic gonadotropin, calcitonin, bladder tumor antigen, CD7, CD19, CD117, CA15-3, CA19-9, CA-125, CA 27.29, CA72-4, CD20, CD22, CD25, CD30, CD33, chromogranin A, cytokeratin fragment 21-1, estrogen receptor, progesterone receptor, fibrinogen, gastrin, HE4, Her2 / neu, neuron-specific enolase, nuclear matrix protein 22, prostatic acid phosphatase, PD-L1, PD-1, prostate-specific antigen, somatostatin receptor, thyroglobulin, 5-HIAA, osteocalcin, transferrin recepto, alkaline phosphtase, BRAF, KRAS, NMP22, BRCA2, urokinase plasminogen activator, apoliprotein A1, S100, nestin, cytokeratin fragments 21-1, ferritin, tissue polypeptide antigen, epididymal secretory protein E4, CECAM 5, CECAM 6, serum M-protein, CTLA-4, B7-1 / B7-2, epithelial tumor antigen, tyrosinase, melanoma- associated antigen, p53, mutated p53, MART-2, beta-catenin, mutated ras, mutated KRAS, BAGE, GAGE-1, NY-ESO-1, PRAME, CT83, SSX2, ART-4, BAGE, beta-catenin / m, Bcr-abL CAMEL, CAP-1 , CASP-8, CDC27 / m, CD 4 / m, a cell surface protein of the claudin family, c- MYC, CT, Cyp-B, DAM, ELF2M, ETV6-AML1, G250, GAGE, GnT-V, KPL-1, HAGE, HPV- E7, HPV-E6, HAST-2, hTERT, LAGE, LDLR / FUT, MAGE- A, MAGE-B, MAGE-C, MART- 1, Melan-A, MCIR-1, myosin, MUM-1, MUM-2, MUM-3, NA88-A, NF1, NY-ESO-1, NY-BR- 1, HSP90, BCR-ABL-1, RAR, PRAME, proteinase 3, PSM, RAGE, RU1, RU2, SAGE, SART- 1, SART-3, SCGB3A2, SCP 1, SCP2, SCP3, SSX, Survrvin, TEL / AML1, TPEM, TRP-1, TRP- 2, TRP-2-INT2, TPTE, WT, or a combination of two or more thereof. Only one type of antigen binding domain may be used, or two or more types thereof may be used in combination.

[0175] In embodiments, the antigen binding domain binds to alpha-fetoprotein. In embodiments, the antigen binding domain binds to carcinoembryonic antigen (CEA). In embodiments, the antigen binding domain binds to beta-2-microglobulin. In embodiments, the antigen binding domain binds to beta-human chorionic gonadotropin. In embodiments, the antigen binding domain binds to calcitonin. In embodiments, the antigen binding domain binds to bladder tumor antigen. In embodiments, the antigen binding domain binds to CD117. In embodiments, the antigen binding domain binds to CA15-3. In embodiments, the antigen binding domain binds to CA19-9. In embodiments, the antigen binding domain binds to CA-125. In embodiments, the antigen binding domain binds to CA 27.29. In embodiments, the antigen binding domain binds to CA72-4, calcitonin. In embodiments, the antigen binding domain binds to carcinoembryonic antigen. In embodiments, the antigen binding domain binds to CD20. In embodiments, the antigen binding domain binds to CD22. In embodiments, the antigen binding domain binds to CD25. In embodiments, the antigen binding domain binds to CD30. In embodiments, the antigen binding domain binds to CD33. In embodiments, the antigen binding domain binds to chromogranin A. In embodiments, the antigen binding domain binds to cytokeratin fragment 21-1. In embodiments, the antigen binding domain binds to estrogen receptor. In embodiments, the antigen binding domain binds to progesterone receptor. In embodiments, the antigen binding domain binds to fibrinogen. In embodiments, the antigen binding domain binds to gastrin. In embodiments, the antigen binding domain binds to HE4. In embodiments, the antigen binding domain binds to Her2 / neu. In embodiments, the antigen binding domain binds to neuron-specific enolase. In embodiments, the antigen binding domain binds to nuclear matrix protein 22. In embodiments, the antigen binding domain binds to prostatic acid phosphatase. In embodiments, the antigen binding domain binds to PD-L1. In embodiments, the antigen binding domain binds to PD-1. In embodiments, the antigen binding domain binds to prostate-specific antigen. In embodiments, the antigen binding domain binds to somatostatin receptor. In embodiments, the antigen binding domain binds to thyroglobulin. In embodiments, the antigen binding domain binds to 5-HIAA. In embodiments, the antigen binding domain binds to osteocalcin. In embodiments, the antigen binding domain binds to transferrin receptor. In embodiments, the antigen binding domain binds to alkaline phosphtase. In embodiments, the antigen binding domain binds to BRAF. In embodiments, the antigen binding domain binds to KRAS. In embodiments, the antigen binding domain binds to NMP22. In embodiments, the antigen binding domain binds to BRCA2. In embodiments, the antigen binding domain binds to urokinase plasminogen activator. In embodiments, the antigen binding domain binds to apoliprotein A1. In embodiments, the antigen binding domain binds to S100. In embodiments, the antigen binding domain binds to nestin. In embodiments, the antigen binding domain binds to cytokeratin fragment 21-1. In embodiments, the antigen binding domain binds to ferritin. In embodiments, the antigen binding domain binds to tissue polypeptide antigen. In embodiments, the antigen binding domain binds to epididymal secretory protein E4. In embodiments, the antigen binding domain binds to CECAM 5. In embodiments, the antigen binding domain binds to CECAM 6. In embodiments, the antigen binding domain binds to serum M-protein. In embodiments, the antigen binding domain binds to CTLA-4. In embodiments, the antigen binding domain binds to B7-1 / B7-2. In embodiments, the antigenbinding domain binds to MUC-1. In embodiments, the antigen binding domain binds to epithelial tumor antigen. In embodiments, the antigen binding domain binds to tyrosinase. In embodiments, the antigen binding domain binds to melanoma-associated antigen. In embodiments, the antigen binding domain binds to p53. In embodiments, the antigen binding domain binds to MART-2. In embodiments, the antigen binding domain binds to beta-catenin. In embodiments, the antigen binding domain binds to an oncofetal protein. In embodiments, the oncofetal protein is alpha-fetoprotein, carcinoembryonic antigen (CEA), trophoblastglycoprotein precursor, or immature laminin receptor protein. In embodiments, the antigenbinding domain binds to mutated ras. In embodiments, the antigen binding domain binds to mutated KRAS. In embodiments, the antigen binding domain binds to a cancer testis (CT) antigen. In embodiments, the antigen binding domain binds to MAGE-A1. In embodiments, the antigen binding domain binds to MAGE-A3. In embodiments, the antigen binding domain binds to MAGE-A4. In embodiments, the antigen binding domain binds to BAGE. In embodiments, the antigen binding domain binds to In embodiments, the antigen binding domain binds to GAGE-1. In embodiments, the antigen binding domain binds to NY-ESO-1. In embodiments, the antigen binding domain binds to PRAME. In embodiments, the antigen binding domain binds to CT83. In embodiments, the antigen binding domain binds to ART-4. In embodiments, the antigen binding domain binds to BAGE. In embodiments, the antigen binding domain binds to beta-catenin. In embodiments, the antigen binding domain binds to Bcr-abL. In embodiments, the antigen binding domain binds to CAMEL. In embodiments, the antigen binding domain binds to CAP-1. In embodiments, the antigen binding domain binds to CASP-8. In embodiments, the antigen binding domain binds to CDC27 / m. In embodiments, the antigen binding domain binds to CD 4 / m. In embodiments, the antigen binding domain binds to CLAUDIN-6. In embodiments, the antigen binding domain binds to CLAUDIN-18.2. In embodiments, the antigen binding domain binds to CLAUDIN-12. In embodiments, the antigen binding domain binds to c-MYC. In embodiments, the antigen binding domain binds to Cyp-B. In embodiments, the antigen binding domain binds to DAM. In embodiments, the antigen binding domain binds to ELF2M. In embodiments, the antigen binding domain binds to ETV6- AML1. In embodiments, the antigen binding domain binds to G250. In embodiments, the antigen binding domain binds to GAGE. In embodiments, the antigen binding domain binds to GnT-V. In embodiments, the antigen binding domain binds to KPL-1. In embodiments, the antigen binding domain binds to HAGE. In embodiments, the antigen binding domain binds to HPV-E7. In embodiments, the antigen binding domain binds to HPV-E6. In embodiments, the antigen binding domain binds to HAST-2. In embodiments, the antigen binding domain binds tohTERT. In embodiments, the antigen binding domain binds to LAGE. In embodiments, the antigen binding domain binds to LDLR / FUT. In embodiments, the antigen binding domain binds to MAGE-A2. In embodiments, the antigen binding domain binds to MAGE-A5. In embodiments, the antigen binding domain binds to MAGE-A6. In embodiments, the antigen binding domain binds to MAGE-A7. In embodiments, the antigen binding domain binds to MAGE-A8. In embodiments, the antigen binding domain binds to MAGE-A9. In embodiments, the antigen binding domain binds to MAGE-A10. In embodiments, the antigen binding domain binds to MAGE-A11. In embodiments, the antigen binding domain binds to MAGE- A12. In embodiments, the antigen binding domain binds to MAGE-B. In embodiments, the antigen binding domain binds to MAGE-C. In embodiments, the antigen binding domain binds to MART-1. In embodiments, the antigen binding domain binds to Melan-A. In embodiments, the antigen binding domain binds to MCIR-1. In embodiments, the antigen binding domain binds to myosin. In embodiments, the antigen binding domain binds to MUM-1. In embodiments, the antigen binding domain binds to MUM-2. In embodiments, the antigen binding domain binds to MUM-3. In embodiments, the antigen binding domain binds to NA88-A. In embodiments, the antigen binding domain binds to NF1. In embodiments, the antigen binding domain binds to NY-ESO-1. In embodiments, the antigen binding domain binds to NY-BR-1. In embodiments, the antigen binding domain binds to HSP90. In embodiments, the antigen binding domain binds to BCR-ABL-1. In embodiments, the antigen binding domain binds to RAR. In embodiments, the antigen binding domain binds to PRAME. In embodiments, the antigen binding domain binds to proteinase 3. In embodiments, the antigen binding domain binds to PSM. In embodiments, the antigen binding domain binds to RAGE. In embodiments, the antigen binding domain binds to RU1. In embodiments, the antigen binding domain binds to RU2. In embodiments, the antigen binding domain binds to SAGE. In embodiments, the antigen binding domain binds to SART-1. In embodiments, the antigen binding domain binds to SART-3. In embodiments, the antigen binding domain binds to SCGB3A2. In embodiments, the antigen binding domain binds to SCP1. In embodiments, the antigen binding domain binds to SCP2. In embodiments, the antigen binding domain binds to SCP3. In embodiments, the antigen binding domain binds to Survivin. In embodiments, the antigen binding domain binds to TEL / AML1. In embodiments, the antigen binding domain binds to TPEM. In embodiments, the antigen binding domain binds to TRP-1. In embodiments, the antigen binding domain binds to TRP-2. In embodiments, the antigen binding domain binds to TRP-2-INT2. In embodiments, the antigen binding domain binds to TPTE. In embodiments, the antigen binding domain binds to WT-1.

[0176] In embodiments, the antigen binding domain binds to CD19 or BCMA. Inembodiments, the antigen binding domain binds to CD19. In embodiments, the antigen binding domain binds to BCMA. In embodiments, the antigen binding domain binds to epidermal growth factor receptor, mesothelin, prostate-specific membrane antigen, carcinoembryonic antigen, disialoganglioside GD2, interleukin-13Ra2, glypican-3, carbonic anhydrase IX, L1CAM, CA-125, CD133, fibroblast activation protein, a cancer testis antigen, MUC1, or FOLR1. In embodiments, the antigen binding domain binds to epidermal growth factor receptor variant III or human epidermal growth factor receptor 2. In embodiments, the antigen binding domain binds to epidermal growth factor receptor. In embodiments, the antigen binding domain binds to mesothelin. In embodiments, the antigen binding domain binds to prostate-specific membrane antigen. In embodiments, the antigen binding domain binds to disialoganglioside GD2. In embodiments, the antigen binding domain binds to interleukin-13Ra2. In embodiments, the antigen binding domain binds to glypican-3. In embodiments, the antigen binding domain binds to carbonic anhydrase IX. In embodiments, the antigen binding domain binds to L1CAM. In embodiments, the antigen binding domain binds to CD133. In embodiments, the antigen binding domain binds to fibroblast activation protein. In embodiments, the antigen binding domain binds to a cancer testis antigen. In embodiments, the antigen binding domain binds to MUC1. In embodiments, the antigen binding domain binds to FOLR1.

[0177] CAR Transmembrane domain. With respect to the transmembrane domain, in embodiments, a CAR can be designed to comprise a transmembrane domain that is attached to the extracellular domain of the CAR. A transmembrane domain can include one or more additional amino acids adjacent to the transmembrane region, e.g., one or more amino acid associated with the extracellular region of the protein from which the transmembrane was derived and / or one or more additional amino acids associated with the intracellular region of the protein from which the transmembrane protein is derived. In embodiments, the transmembrane domain is one that is associated with one of the other domains of the CAR. In embodiments, the transmembrane domain can be selected or modified by amino acid substitution to avoid binding of such domains to the transmembrane domains of the same or different surface membrane proteins, e.g., to minimize interactions with other members of the receptor complex. In embodiments, the transmembrane domain is capable of homodimerization with another CAR on the cell surface of a CAR T-cell. In embodiments, the amino acid sequence of the transmembrane domain may be modified or substituted so as to minimize interactions with the binding domains of the native binding partner present in the same CAR T-cell.

[0178] The transmembrane domain may be derived either from a natural or from a recombinant source. Where the source is natural, the domain may be derived from anymembrane-bound or transmembrane protein. In embodiments, the transmembrane domain is capable of signaling to the intracellular domain(s) whenever the CAR has bound to a target. A transmembrane domain may include at least the transmembrane region(s) of e.g., the alpha, beta or zeta chain of the T-cell receptor, CD28, CD27, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, CD154. In embodiments, a transmembrane domain may include at least the transmembrane region(s) of, e.g., KIR2DS2, OX40, CD2, CD27, LFA-1 (CD11a, CD18), ICOS (CD278), 4-1BB (CD137), GITR, CD40, BAFFR, HVEM (LIGHTR), SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD160, CD19, IL2R beta, IL2R gamma, IL7R α, ITGA1, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, TNFR2, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, PAG / Cbp, NKG2D, NKG2C. In embodiments, the transmembrane domain can be attached to the extracellular region of the CAR, e.g., the antigen binding domain of the CAR, via a hinge, e.g., a hinge from a human protein. For example, in one embodiment, the hinge can be a human Ig (immunoglobulin) hinge, e.g., an IgG4 hinge, or a CD8a hinge.

[0179] Intracellular or Cytoplasmic domain. The intracellular signaling domain of a CAR isresponsible for activation of at least one of the normal effector functions of the T cell in which the chimeric receptor has been placed. The term “effector function” refers to a specialized function of a differentiated cell. Effector function of a T cell, for example, may be cytolytic activity or helper activity including the secretion of cytokines. Effector function in a naive, memory, or memory-type T cell includes antigen-dependent proliferation. Thus, the term “intracellular signaling domain” refers to the portion of a protein that transduces the effector function signal and directs the cell to perform a specialized function. While usually the entire intracellular signaling domain is employed, in many cases it will not be necessary to use the entire intracellular polypeptide. To the extent that a truncated portion of the intracellular signaling domain may find use, such truncated portion may be used in place of the intact chain as long as it still transduces the effector function signal. The term intracellular signaling domain is thus meant to include any truncated portion of the intracellular signaling domain sufficient to transduce the effector function signal.

[0180] Examples of intracellular signaling domains include, but are not limited to, cytoplasmic sequences of the T cell receptor (TCR) and co-receptors that act in concert toinitiate signal transduction following antigen receptor engagement, as well as any derivative or variant of these sequences and any synthetic sequence that has the same functional capability. It is known that signals generated through the TCR alone are insufficient for full activation of the T cell and that a secondary or co-stimulatory signal is also required. Thus, T cell activation can be mediated by two distinct classes of cytoplasmic signaling sequence: those that initiate antigen-dependent primary activation through the TCR (primary cytoplasmic signaling sequences) and those that act in an antigen-independent manner to provide a secondary or co- stimulatory signal (secondary cytoplasmic signaling sequences).

[0181] In embodiments, the CAR-T cell described herein can further comprise a second CAR, e.g., a second CAR that includes a different antigen binding domain, e.g., to the same target or a different target (e.g., a target other than a cancer associated antigen described herein or a different cancer associated antigen described herein, e.g., CD19, CD33, CLL-1, CD34, FLT3, folate receptor alpha, or folate receptor beta). In embodiments, the second CAR includes an antigen binding domain to a target expressed the same cancer cell type as the cancer associated antigen. In embodiments, the CAR-expressing cell comprises a first CAR that targets a first antigen and includes an intracellular signaling domain having a costimulatory signaling domain but not a primary signaling domain, and a second CAR that targets a second, different, antigen and includes an intracellular signaling domain having a primary signaling domain but not a costimulatory signaling domain. While not wishing to be bound by theory, placement of a costimulatory signaling domain, e.g., 4-1BB, CD28, ICOS, CD27 or OX-40, onto the first CAR, and the primary signaling domain, e.g., CD3 zeta, on the second CAR can limit the CAR activity to cells where both targets are expressed. In one embodiment, the CAR expressing cell comprises a first cancer associated antigen CAR that includes an antigen binding domain that binds a target antigen described herein, a transmembrane domain and a costimulatory domain and a second CAR that targets a different target antigen (e.g., an antigen expressed on that same cancer cell type as the first target antigen) and includes an antigen binding domain, a transmembrane domain and a primary signaling domain. In embodiments, the CAR expressing cell comprises a first CAR that includes an antigen binding domain that binds a target antigen described herein, a transmembrane domain and a primary signaling domain and a second CAR that targets an antigen other than the first target antigen (e.g., an antigen expressed on the same cancer cell type as the first target antigen) and includes an antigen binding domain to the antigen, a transmembrane domain and a costimulatory signaling domain.

[0182] In embodiments, the disclosure provides a population of CAR-T cells. In embodiments, the population of CAR T-cells comprises a mixture of cells expressing differentCARs. In embodiments, the population of CART cells can include a first cell expressing a CAR having an antigen binding domain to a cancer associated antigen described herein, and a second cell expressing a CAR having a different antigen binding domain, e.g., an antigen binding domain to a different a cancer associated antigen described herein, e.g., an antigen binding domain to a cancer associated antigen described herein that differs from the cancer associate antigen bound by the antigen binding domain of the CAR expressed by the first cell. As another example, the population of CAR T-cells can include a first cell expressing a CAR that includes an antigen binding domain to a cancer associated antigen described herein, and a second cell expressing a CAR that includes an antigen binding domain to a target other than a cancer associate antigen as described herein. In the population of CAR T-cells includes, e.g., a first cell expressing a CAR that includes a primary intracellular signaling domain, and a second cell expressing a CAR that includes a secondary signaling domain.

[0183] In embodiments, the disclosure provides a population of cells wherein at least one cell in the population expresses a CAR having an antigen binding domain to a cancer associated antigen described herein, and a second cell expressing another agent, e.g., an agent which enhances the activity of a CAR-expressing cell. For example, the agent can be an agent which inhibits an inhibitory molecule. Inhibitory molecules, e.g., PD-1, can, in embodiments, decrease the ability of a CAR T-cells to mount an immune effector response. Examples of inhibitory molecules include PD-1, PD-L1, CTLA4, TIM3, CEACAM (CEACAM-1, CEACAM-3, and / or CEACAM-5), LAG3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4, CD80, CD86, B7-H3 (CD276), B7-H4 (VTCN1), HVEM (TNFRSF14 or CD270), KIR, A2aR, MHC class I, MHC class II, GAL9, adenosine, and TGF (e.g., TGFbeta). In embodiments, the agent which inhibits an inhibitory molecule comprises a first polypeptide, e.g., an inhibitory molecule, associated with a second polypeptide that provides a positive signal to the cell, e.g., an intracellular signaling domain described herein. In one embodiment, the agent comprises a first polypeptide, e.g., of an inhibitory molecule such as PD-1, PD-L1, CTLA4, TIM3, CEACAM (CEACAM-1, CEACAM-3, and / or CEACAM-5), LAG3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and TGF beta, or a fragment of any of these, and a second polypeptide which is an intracellular signaling domain described herein (e.g., comprising a costimulatory domain (e.g., 41BB, CD27, OX40 or CD28, e.g., as described herein) and / or a primary signaling domain (e.g., a CD3 zeta signaling domain described herein). In one embodiment, the agent comprises a first polypeptide of PD-1 or a fragment thereof, and a second polypeptide of an intracellular signaling domain described herein (e.g., a CD28 signaling domain described herein and / or a CD3 zeta signaling domain described herein).

[0184] Production of CAR T-cells

[0185] CAR T-cell therapy is a type of immunotherapy that involves modifying a patient’s T- cells to express a synthetic receptor called a chimeric antigen receptor (CAR). These modified CAR T-cells are then infused back into the patient to target and destroy cancer cells. The process involves several steps as follows.1. Collection of T cells: T cells are usually collected from the patient’s own blood through a process called leukapheresis (i.e., autologous T cells). In some cases, T cells from a donor may be used (i.e., allogeneic T cells). The T-cells can be CD4+T- cells and / or CD8+T-cells.2. Isolation and purification: The collected T cells are isolated and purified from other blood components.3. Genetic modification: The isolated T cells are genetically modified to express the CAR. This is typically done using viral vectors, often lentiviruses or retroviruses, to introduce the CAR into the T cells. For example, the T cells can be genetically modified to express the CD19-specific CAR. The CD19-specific CAR is designed to recognize and bind to the CD19 antigen, which is commonly found on the surface of B cells, including B-cell malignancies (e.g., B cell lymphomas). The CAR construct includes a promoter to drive the expression of the CAR and signaling domain, typically derived from the T-cell receptor or co-stimulatory molecules, to activate the T cells upon binding to CD19. 4. Expansion. The genetically modified T cells are then cultured and expanded in the laboratory to increase their numbers. This step is crucial to generate a sufficient quantity of CAR T-cells for the therapeutic dose.5. Quality control: The CAR T-cells undergo rigorous testing to ensure they meet quality and safety standards. This includes assessing the level of CAR expression, testing for contaminants, and confirming the absence of replication-competent virus.6. Condition: Before the CAR T-cells are infused back into the patient, the patient my undergo a conditioning regimen, which may involve chemotherapy, to create an environment conducive to the CAR T-cells’ activity.7. Infusion: The modified CAR T cells are infused back into the patient through a vein, similar to a blood transfusion.8. Monitoring and follow-up: Patients are closely monitored for potential side effects, and follow-up care is provided to manage any adverse reactions. The persistence and effectiveness of the CAR T-cells in targeting and destroying cancer cells are also monitored over time.

[0186] In the methods described herein, the patient can be administered an effective amount of an ITK inhibitor and an effective amount of CAR T-cells. In embodiments, the ITK inhibitor is administered to the patient before the CAR T-cells are administered to the patient. In embodiments, the ITK inhibitor is administered to the patient before and after the CAR T-cells are administered to the patient. In embodiments, the ITK inhibitor is administered to the patient after the CAR T-cells are administered to the patient. In embodiments, the ITK inhibitor andCAR T-cells are concomitantly administered to the patient.

[0187] The term “concomitantly” or “concomitant” refers to the administration of the ITK inhibitor and CAR T-cells about the same time. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 24 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 12 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 10 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 8 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 6 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 4 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 3 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 2 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 1 hour of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 30 minutes of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 15 minutes of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 1 minute to about 24 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 1 minute to about 12 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 1 minute to about 10 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 1 minute to about 6 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 1 minute to about 5 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 1 minute to about 4 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 1 minute to about 3 hours of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 1 minute to about 2 hours of each other. Inembodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 1 minute to about 1 hour of each other. In embodiments, concomitant administration refers to the administration of the ITK inhibitor and CAR T-cells within about 1 minute to about 30 minutes of each other.

[0188] The term “before” with reference to the administration of the ITK inhibitor before the CAR T-cells (or vice versa) means that the ITK inhibitor is administered at least 24 hours before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered at least 2 days before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered at least 3 days before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered at least 4 days before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered at least 5 days before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered at least 6 days before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered at least 7 days before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered at least 2 weeks before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered at least 3 weeks before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered at least 4 weeks before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered from about 24 hours to about 4 weeks before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered from about 24 hours to about 3 weeks before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered from about 24 hours to about 2 weeks before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered from about 24 hours to about 7 days before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered from about 24 hours to about 6 days before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered from about 24 hours to about 7 days before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered from about 24 hours to about 4 days before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered from about 24 hours to about 3 days before the CAR T-cells. In embodiments, before means that the ITK inhibitor is administered from about 24 hours to about 2 days before the CAR T-cells.

[0189] The term “after” with reference to the administration of the ITK inhibitor after the CAR T-cells means that the ITK inhibitor is administered at least 24 hours after the CAR T- cells. In embodiments, after means that the ITK inhibitor is administered at least 2 days after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered at least 3 daysafter the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered at least 4 days after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered at least 5 days after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered at least 6 days after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered at least 7 days after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered at least 2 weeks after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered at least 3 weeks after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered at least 4 weeks after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered from about 24 hours to about 4 weeks after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered from about 24 hours to about 3 weeks after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered from about 24 hours to about 2 weeks after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered from about 24 hours to about 7 days after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered from about 24 hours to about 6 days after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered from about 24 hours to about 7 days after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered from about 24 hours to about 4 days after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered from about 24 hours to about 3 days after the CAR T-cells. In embodiments, after means that the ITK inhibitor is administered from about 24 hours to about 2 days after the CAR T-cells.

[0190] Dose and Dosing Regimens

[0191] The dosage, route of administration, and frequency of administration of the ITK inhibitors and CAR T-cells administered to a subject can vary depending upon a variety of factors, for example, whether the mammal suffers from another disease, and its route of administration; size, age, sex, health, body weight, body mass index, and diet of the recipient; nature and extent of symptoms of the disease being treated, kind of concurrent treatment, complications from the disease being treated or other health-related problems. Other therapeutic regimens or agents can be used in conjunction with the methods, ITK inhibitors, and CAR T- cells described herein. Adjustment and manipulation of established dosages (e.g., frequency, route, and duration) are well within the ability of those skilled in the art.

[0192] For any composition, ITK inhibitor, and CAR T-cells described herein, the effective amount can be initially determined from cell culture assays. Target concentrations will be those concentrations of ITK inhibitors and CAR T-cells that are capable of achieving the methodsdescribed herein, as measured using the methods described herein or known in the art. As is known in the art, effective amounts of ITK inhibitors and CAR T-cells for use in humans can also be determined from animal models. For example, a dose for humans can be formulated to achieve a concentration that has been found to be effective in animals. The dosage in humans can be adjusted by monitoring effectiveness and adjusting the dosage upwards or downwards, as described above. Adjusting the dose to achieve maximal efficacy in humans based on the methods described above and other methods is well within the capabilities of the ordinarily skilled artisan.

[0193] Dosages of the ITK inhibitors and CAR T-cells may be varied depending upon the requirements of the patient. The dose administered to a patient should be sufficient to affect a beneficial therapeutic response in the patient over time. The size of the dose also will be determined by the existence, nature, and extent of any adverse side-effects. Determination of the proper dosage for a particular situation is within the skill of the art. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the ITK inhibitor and CAR T-cells. Thereafter, the dosage is increased by small increments until the optimum effect under circumstances is reached. Dosage amounts and intervals can be adjusted individually to provide levels of the ITK inhibitors and CAR T-cells effective for the particular clinical indication being treated. This will provide a therapeutic regimen that is commensurate with the severity of the individual's disease state.

[0194] In embodiments of the methods described herein, the effective amount of the CAR T- cells is from 103to 1010cells per kg body weight of the human patient. In embodiments, the effective amount of the CAR T-cells is from 104to 109cells per kg body weight of the human patient. In embodiments, the effective amount of the CAR T-cells is from 105to 106cells per kg body weight of the human patient.

[0195] In embodiments of the methods described herein, the effective amount is about 50 mg to about 1,000 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 100 mg to about 1,000 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 250 mg to about 950 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 250 mg to about 900 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 250 mg to about 850 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 250 mg to about 800 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 250 mg to about 750 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 250 mg to about 700 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 250 mg to about 650 mg of the ITKinhibitor per day. In embodiments, the effective amount is about 250 mg to about 600 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 250 mg to about 550 mg of the ITK inhibitor per day.

[0196] In embodiments of the methods described herein, the effective amount is about 300 mg to about 1,000 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 300 mg to about 950 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 300 mg to about 900 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 300 mg to about 850 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 300 mg to about 800 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 300 mg to about 750 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 300 mg to about 700 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 300 mg to about 650 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 300 mg to about 600 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 300 mg to about 550 mg of the ITK inhibitor per day.

[0197] In embodiments of the methods described herein, the effective amount is about 300 mg to about 500 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 305 mg to about 495 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 310 mg to about 490 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 315 mg to about 485 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 320 mg to about 480 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 325 mg to about 475 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 330 mg to about 470 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 335 mg to about 465 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 340 mg to about 460 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 345 mg to about 455 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 350 mg to about 450 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 355 mg to about 445 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 360 mg to about 440 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 365 mg to about 435 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 370 mg to about 430 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 375 mg to about 425 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 380 mg to about 420 mg of the ITK inhibitor per day. In embodiments, the effective amount isabout 385 mg to about 415 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 390 mg to about 410 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 395 mg to about 405 mg of the ITK inhibitor per day. In embodiments, the effective amount is about 400 mg of the ITK inhibitor per day.

[0198] The term “per day” encompasses once per day, twice per day, three times per day, or four times per day. In embodiments, “per day” is once per day. In embodiments, “per day” is twice per day. In embodiments, “per day” is thrice per day. In embodiments, “per day” is four times per day. For example, if the effective of the amount of the ITK inhibitor is 400 mg per day, then the ITK inhibitor can be administered in an amount of 400 mg once per day, 200 mg twice per day, or 100 mg four times per day.

[0199] In embodiments of the methods described herein, the effective amount is about 125 mg to about 500 mg of the ITK inhibitor twice per day (BID). In embodiments, the effective amount is about 125 mg to about 475 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 125 mg to about 450 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 125 mg to about 425 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 125 mg to about 400 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 125 mg to about 375 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 125 mg to about 350 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 125 mg to about 325 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 125 mg to about 300 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 125 mg to about 275 mg of the ITK inhibitor twice per day.

[0200] In embodiments of the methods described herein, the effective amount is about 150 mg to about 500 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 150 mg to about 475 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 150 mg to about 450 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 150 mg to about 425 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 150 mg to about 400 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 150 mg to about 375 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 150 mg to about 350 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 150 mg to about 325 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 150 mg to about 300 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 150 mg to about 275 mg of the ITK inhibitor twice per day.

[0201] In embodiments of the methods described herein, the effective amount is about 150 mg to about 250 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 152.5 mg to about 247.5 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 155 mg to about 245 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 157.5 mg to about 242.5 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 160 mg to about 240 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 162.5 mg to about 237.5 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 165 mg to about 235 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 167.5 mg to about 232.5 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 170 mg to about 230 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 172.5 mg to about 227.5 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 175 mg to about 225 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 177.5 mg to about 222.5 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 180 mg to about 220 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 182.5 mg to about 217.5 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 185 mg to about 215 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 187.5 mg to about 212.5 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 190 mg to about 210 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 192.5 mg to about 207.5 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 195 mg to about 205 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 197.5 mg to about 202.5 mg of the ITK inhibitor twice per day. In embodiments, the effective amount is about 200 mg of the ITK inhibitor twice per day.

[0202] Pharmaceutical Compositions

[0203] Provided herein are pharmaceutical compositions comprising an ITK inhibitor and a pharmaceutically acceptable excipient. The compositions are suitable for formulation and administration in vitro or in vivo. Suitable carriers and excipients and their formulations are described in Remington: The Science and Practice of Pharmacy, 21st Edition, David B. Troy, ed., Lippicott Williams & Wilkins (2005).

[0204] “Pharmaceutically acceptable excipient” and “pharmaceutically acceptable carrier” refer to a substance that aids the administration of an active agent to and absorption by a subject and can be included in the compositions of the disclosure without causing a significant adversetoxicological effect on the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline solutions, lactated Ringer’s, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer's solution), alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethycellulose, polyvinyl pyrrolidine, and colors, and the like. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and / or aromatic substances and the like that do not deleteriously react with the compounds of the disclosure. One of skill in the art will recognize that other pharmaceutical excipients are useful.

[0205] Solutions of the active compounds as free base or pharmacologically acceptable salt can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations can contain a preservative to prevent the growth of microorganisms.

[0206] Pharmaceutical compositions can be delivered via intranasal or inhalable solutions or sprays, aerosols or inhalants. Nasal solutions can be aqueous solutions designed to be administered to the nasal passages in drops or sprays. Nasal solutions can be prepared so that they are similar in many respects to nasal secretions. Thus, the aqueous nasal solutions usually are isotonic and slightly buffered to maintain a pH of 5 to 7. In addition, antimicrobial preservatives, similar to those used in ophthalmic preparations and appropriate drug stabilizers, if required, may be included in the formulation. Various commercial nasal preparations are known and can include, for example, antibiotics and antihistamines.

[0207] Oral formulations can include excipients as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate and the like. These compositions take the form of solutions, suspensions, tablets, pills, capsules, sustained release formulations or powders. In embodiments, oral pharmaceutical compositions will comprise an inert diluent or edible carrier, or they may be enclosed in hard or soft shell gelatin capsule, or they may be compressed into tablets, or they may be incorporated directly with the food. For oral therapeutic administration, the active compounds may be incorporated with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. The percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 1 to about 90% of the weight of the unit. The amount of active compounds in such compositions is suchthat a suitable dosage can be obtained.

[0208] For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered and the liquid diluent first rendered isotonic with sufficient saline or glucose. Aqueous solutions, in particular, sterile aqueous media, are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. For example, one dosage could be dissolved in 1 ml of isotonic NaCl solution and either added to 1000 ml of hypodermoclysis fluid or injected at the proposed site of infusion.

[0209] Sterile injectable solutions can be prepared by incorporating the active compounds in the required amount in the appropriate solvent followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium. Vacuum-drying and freeze-drying techniques, which yield a powder of the active ingredient plus any additional desired ingredients, can be used to prepare sterile powders for reconstitution of sterile injectable solutions. The preparation of more, or highly, concentrated solutions for direct injection is also contemplated. Dimethyl sulfoxide can be used as solvent for extremely rapid penetration, delivering high concentrations of the active agents to a small area.

[0210] The formulations of compounds can be presented in unit-dose or multi-dose sealed containers, such as ampules and vials. Thus, the composition can be in unit dosage form. In such form the preparation is subdivided into unit doses containing appropriate quantities of the active component. Thus, the compositions can be administered in a variety of unit dosage forms depending upon the method of administration. For example, unit dosage forms suitable for oral administration include, but are not limited to, powder, tablets, pills, capsules and lozenges.

[0211] In embodiments, the disclosure provides orally-administrable pharmaceutical compositions comprising an ITK inhibitor and a pharmaceutically acceptable excipient. In embodiments, the pharmaceutical composition is an oral composition. In embodiments, the oral composition is a solid oral composition. In embodiments, the oral composition is a liquid oral composition. In embodiments, the pharmaceutical composition is a tablet or a capsule. In embodiments, the pharmaceutical composition is a tablet. In embodiments, the pharmaceutical composition is a capsule. In embodiments, the pharmaceutical composition is a powder.

[0212] Embodiments

[0213] Embodiment 1. A method of treating cancer in a patient in need thereof, the method comprising administering to the patient: (i) an effective amount of an interleukin-2-inducible T- cell kinase inhibitor and (ii) an effective amount of CAR T-cells, thereby treating cancer in thepatient.

[0214] Embodiment 2. A method of treating cancer in a patient in need thereof, the method comprising: (i) contacting CAR T-cells with an effective amount of an interleukin-2-inducible T-cell kinase inhibitor in vitro, thereby producing ITK-contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK-contacted CAR T-cells; thereby treating cancer in the patient.

[0215] Embodiment 3. The method of embodiment 2, further comprising administering to the patient an effective amount of the interleukin-2-inducible T-cell kinase inhibitor.

[0216] Embodiment 4. The method of any one of embodiments 1 to 3, wherein the cancer is lymphoma, leukemia, lung cancer, colorectal cancer, pancreatic cancer, prostate cancer, breast cancer, gastric cancer, renal cancer, melanoma, or head and neck cancer.

[0217] Embodiment 5. The method of any one of embodiments 1 to 3, wherein the cancer isB-cell precursor acute lymphoblastic leukemia, mantle cell lymphoma, follicular lymphoma, ormultiple myeloma.

[0218] Embodiment 6. The method of any one of embodiments 1 to 3, wherein the cancer is lymphoma.

[0219] Embodiment 7. The method of embodiment 6, wherein the lymphoma is T-cell lymphoma.

[0220] Embodiment 8. The method of embodiment 6, wherein the lymphoma is B-cell lymphoma.

[0221] Embodiment 9. The method of embodiment 8, wherein the B-cell lymphoma is diffuse large B-cell lymphoma or high grade B-cell lymphoma.

[0222] Embodiment 10. A method for preventing and / or reversing T-cell exhaustion in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2-inducible T-cell kinase inhibitor; thereby preventing and / or reversing T-cell exhaustion in the patient.

[0223] Embodiment 11. The method of embodiment 10, wherein the T-cell is a CAR T-cell.

[0224] Embodiment 12. A method for preventing and / or reversing CAR T-cell exhaustion in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2-inducible T-cell kinase inhibitor; wherein the patient is receiving or has received CAR T-cell therapy; thereby preventing and / or reversing CAR T-cell exhaustion in the patient.

[0225] Embodiment 13. A method of preventing and / or reversing CAR T-cell exhaustion in apatient in need thereof, the method comprising administering to the patient: (i) an effective amount of an interleukin-2-inducible T-cell kinase inhibitor and (ii) an effective amount of CAR T-cells, thereby preventing and / or reducing CAR T-cell exhaustion in the patient.

[0226] Embodiment 14. A method of preventing and / or reversing CAR T-cell exhaustion in a patient in need thereof, the method comprising: (i) contacting CAR T-cells with an effective amount of an interleukin-2-inducible T-cell kinase inhibitor in vitro, thereby producing ITK- contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK- contacted CAR T-cells; thereby preventing and / or reducing CAR T-cell exhaustion in the patient.

[0227] Embodiment 15. The method of embodiment 14, further comprising administering to the patient an effective amount of the interleukin-2-inducible T-cell kinase inhibitor.

[0228] Embodiment 16. A method for stimulating a T-cell-mediated immune response in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2-inducible T-cell kinase inhibitor; thereby stimulating the T-cell-mediated immune response in the patient.

[0229] Embodiment 17. The method of embodiment 16, wherein the T-cell is a CAR T-cell.

[0230] Embodiment 18. A method for stimulating a CAR T-cell-mediated immune response in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2-inducible T-cell kinase inhibitor; wherein the patient is receiving or has received CAR T-cell therapy; thereby stimulating the CAR T-cell-mediated immune response in the patient.

[0231] Embodiment 19. A method for stimulating a CAR T-cell-mediated immune response in a patient in need thereof, the method comprising administering to the patient: (i) an effective amount of an interleukin-2-inducible T-cell kinase inhibitor and (ii) an effective amount of CAR T-cells, thereby stimulating the CAR T-cell-mediated immune response in the patient.

[0232] Embodiment 20. A method for stimulating a CAR T-cell-mediated immune response in a patient in need thereof, the method comprising: (i) contacting CAR T-cells with an effective amount of an interleukin-2-inducible T-cell kinase inhibitor in vitro, thereby producing ITK- contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK- contacted CAR T-cells, thereby stimulating the CAR T-cell-mediated immune response in the patient.

[0233] Embodiment 21. The method of embodiment 20, further comprising administering to the patient an effective amount of the interleukin-2-inducible T-cell kinase inhibitor.

[0234] Embodiment 22. A method for increasing T-cell function in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2- inducible T-cell kinase inhibitor; thereby increasing T-cell function in the patient.

[0235] Embodiment 23. The method of embodiment 22, wherein the T-cell is a CAR T-cell.

[0236] Embodiment 24. A method for increasing CAR T-cell function in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2-inducible T-cell kinase inhibitor; wherein the patient is receiving or has received CAR T-cell therapy; thereby increasing CAR T-cell function in the patient.

[0237] Embodiment 25. A method for increasing CAR T-cell function in a patient in need thereof, the method comprising administering to the patient: (i) an effective amount of an interleukin-2-inducible T-cell kinase inhibitor and (ii) an effective amount of CAR T-cells, thereby increasing CAR T-cell function in the patient.

[0238] Embodiment 26. A method for increasing CAR T-cell function in a patient in need thereof, the method comprising: (i) contacting CAR T-cells with an effective amount of an interleukin-2-inducible T-cell kinase inhibitor in vitro, thereby producing ITK-contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK-contacted CAR T- cells; thereby increasing CAR T-cell function in the patient.

[0239] Embodiment 27. The method of embodiment 26, further comprising administering to the patient an effective amount of the interleukin-2-inducible T-cell kinase inhibitor.

[0240] Embodiment 28. The method of any one of embodiments 22 to 26, wherein the T-cell function is T-cell cytolytic function.

[0241] Embodiment 29. A method of increasing expression of T cell factor-1 on T-cells in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2-inducible T-cell kinase inhibitor; thereby increasing expression of T cell factor-1 on T-cells in the patient.

[0242] Embodiment 30. The method of embodiment 29, wherein the T-cells are CAR T-cells.

[0243] Embodiment 31. A method for increasing expression of T cell factor-1 on CAR T-cells in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2-inducible T-cell kinase inhibitor; wherein the patient is receiving or has received CAR T-cell therapy; thereby increasing expression of T cell factor-1 on CAR T- cells in the patient.

[0244] Embodiment 32. A method of increasing expression of T cell factor-1 on CAR T-cells in a patient in need thereof, the method comprising administering to the patient (i) an effectiveamount of an interleukin-2-inducible T-cell kinase inhibitor and (ii) an effective amount of CAR T-cells, thereby increasing expression of T cell factor-1 on CAR T-cells in the patient.

[0245] Embodiment 33. A method for increasing expression of T-cell factor-1 on CAR T-cells in a patient in need thereof, the method comprising: (i) contacting CAR T-cells with an effective amount of an interleukin-2-inducible T-cell kinase inhibitor in vitro, thereby producing ITK-contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK- contacted CAR T-cells; thereby increasing expression of T-cell factor-1 on CAR T-cells in the patient.

[0246] Embodiment 34. The method of embodiment 33, further comprising administering to the patient an effective amount of the interleukin-2-inducible T-cell kinase inhibitor.

[0247] Embodiment 35. The method of any one of embodiments 1 to 34, further comprising measuring an increased level of TIM3, an increased level of LAG3, an increased level of TIGIT, an increased level of PD-1, a decreased level of IFNγ, a decreased level of granzyme B, or a combination of two or more thereof, relative to a control, in a biological sample obtained from the patient.

[0248] Embodiment 36. The method of any one of embodiments 1 to 35, wherein a biological sample obtained from the patient has an increased level of TIM3, an increased level of LAG3, an increased level of TIGIT, an increased level of PD-1, a decreased level of IFNγ, a decreased level of granzyme B, or a combination of two or more thereof, relative to a control.

[0249] Embodiment 37. The method of any one of embodiment 10 to 36, wherein the patient has cancer.

[0250] Embodiment 38. The method of embodiment 37, wherein the cancer is lymphoma, leukemia, lung cancer, colorectal cancer, pancreatic cancer, prostate cancer, breast cancer, gastric cancer, renal cancer, melanoma, or head and neck cancer.

[0251] Embodiment 39. The method of embodiment 37, wherein the cancer is B-cell precursoracute lymphoblastic leukemia, mantle cell lymphoma, follicular lymphoma, or multiplemyeloma.

[0252] Embodiment 40. The method of embodiment 37, wherein the cancer is lymphoma.

[0253] Embodiment 41. The method of embodiment 40, wherein the lymphoma is T-cell lymphoma.

[0254] Embodiment 42. The method of embodiment 40, wherein the lymphoma is B-cell lymphoma.

[0255] Embodiment 43. The method of embodiment 42, wherein the B-cell lymphoma isdiffuse large B-cell lymphoma or high grade B-cell lymphoma.

[0256] Embodiment 44. A method for making CAR T-cells, the method comprising contacting an interleukin-2-inducible T-cell kinase inhibitor with CAR T-cells in vitro.

[0257] Embodiment 45. A method for increasing expression of T-cell factor-1 on CAR T- cells, the method comprising contacting an interleukin-2-inducible T-cell kinase inhibitor with CAR T-cells in vitro, thereby increasing expression of T-cell factor-1 on CAR T-cells.

[0258] Embodiment 46. A method for increasing CAR T-cell function, the method comprising contacting an interleukin-2-inducible T-cell kinase inhibitor with CAR T-cells in vitro, thereby increasing CAR T-cell function.

[0259] Embodiment 47. The method of embodiment 46, wherein the CAR T-cell function is CAR T-cell cytolytic function.

[0260] Embodiment 48. A method of reducing CAR T-cell exhaustion, the method comprising contacting an interleukin-2-inducible T-cell kinase inhibitor with CAR T-cells in vitro, thereby reducing CAR T-cell exhaustion.

[0261] Embodiment 49. The method of any one of embodiments 1 to 48, wherein the CAR T- cells are CD4+ T-cells.

[0262] Embodiment 50. The method of any one of embodiments 1 to 48, wherein the CAR T- cells are CD8+ T-cells.

[0263] Embodiment 51. The method of any one of embodiments 1 to 48, wherein the CAR T- cells are CD4+ T-cells and CD8+ T-cells.

[0264] Embodiment 52. The method of any one of embodiments 1 to 51, wherein the CAR T- cells are autologous.

[0265] Embodiment 53. The method of any one of embodiments 1 to 51, wherein the CAR T- cells are allogeneic.

[0266] Embodiment 54. The method of any one of embodiments 1 to 53, wherein the CAR T- cells express a chimeric antigen receptor comprising an extracellular domain that binds to an antigen on a cancer cell, wherein the antigen is epidermal growth factor receptor, mesothelin, prostate-specific membrane antigen, disialoganglioside GD2, interleukin-13Ra2, glypican-3, carbonic anhydrase IX, L1CAM, fibroblast activation protein, a cancer testis antigen, MUC1, FOLR1, BCMA, CD133, alpha-fetoprotein, carcinoembryonic antigen, trophoblast glycoproteinprecursor, immature laminin receptor protein, fetal sulphoglycoprotein, g-fetoprotein, cc2-H-ferroportin, beta-2-microglobulin, beta-human chorionic gonadotropin, calcitonin, bladder tumor antigen, CD7, CD19, CD117, CA15-3, CA19-9, CA-125, CA 27.29, CA72-4, CD20, CD22,CD25, CD30, CD33, chromogranin A, cytokeratin fragment 21-1, estrogen receptor, progesterone receptor, fibrinogen, gastrin, HE4, Her2 / neu, neuron-specific enolase, nuclear matrix protein 22, prostatic acid phosphatase, PD-L1, PD-1, prostate-specific antigen, somatostatin receptor, thyroglobulin, 5-HIAA, osteocalcin, transferrin recepto, alkaline phosphtase, BRAF, KRAS, NMP22, BRCA2, urokinase plasminogen activator, apoliprotein A1, S100, nestin, cytokeratin fragments 21-1, ferritin, tissue polypeptide antigen, epididymal secretory protein E4, CECAM 5, CECAM 6, serum M-protein, CTLA-4, B7-1 / B7-2, epithelial tumor antigen, tyrosinase, melanoma-associated antigen, p53, mutated p53, MART-2, beta- catenin, mutated ras, mutated KRAS, BAGE, GAGE-1, NY-ESO-1, PRAME, CT83, SSX2, ART-4, BAGE, beta-catenin / m, Bcr-abL CAMEL, CAP-1 , CASP-8, CDC27 / m, CD 4 / m, a cell surface protein of the claudin family, c-MYC, CT, Cyp-B, DAM, ELF2M, ETV6-AML1, G250, GAGE, GnT-V, KPL-1, HAGE, HPV-E7, HPV-E6, HAST-2, hTERT, LAGE, LDLR / FUT, MAGE- A, MAGE-B, MAGE-C, MART-1, Melan-A, MCIR-1, myosin, MUM-1, MUM-2, MUM-3, NA88-A, NF1, NY-ESO-1, NY-BR-1, HSP90, BCR-ABL-1, RAR, PRAME, proteinase 3, PSM, RAGE, RU1, RU2, SAGE, SART-1, SART-3, SCGB3A2, SCP 1, SCP2, SCP3, SSX, Survrvin, TEL / AML1, TPEM, TRP-1, TRP-2, TRP-2-INT2, TPTE, WT, or a combination of two or more thereof.

[0267] Embodiment 55. The method of any one of embodiments 1 to 53, wherein the CAR T- cells express a chimeric antigen receptor comprising an extracellular domain that binds to an antigen on a cancer cell, wherein the antigen is CD19 or BCMA.

[0268] Embodiment 56. The method of any one of embodiments 1 to 53, wherein the CAR T- cells express a chimeric antigen receptor comprising an extracellular domain that binds to an antigen on a cancer cell, wherein the antigen is epidermal growth factor receptor, mesothelin, prostate-specific membrane antigen, carcinoembryonic antigen, disialoganglioside GD2, interleukin-13Ra2, glypican-3, carbonic anhydrase IX, L1CAM, CA-125, CD133, fibroblast activation protein, a cancer testis antigen, MUC1, or FOLR1.

[0269] Embodiment 57. The method of embodiment 56, wherein the epidermal growth factor receptor is epidermal growth factor receptor variant III or human epidermal growth factor receptor 2.

[0270] Embodiment 58. The method of any one of embodiments 1 to 53, wherein the CAR T- cells are a CAR T-cell therapy selected from the group consisting of axicabtagene ciloleucel,brexucabtagene autoleucel, ciltacabtagene autoleucel, idecabtagene vicleucel, lisocabtagenemaraleucel, and tisagenlecleucel.

[0271] Embodiment 59. The method of any one of embodiments 1 to 58, wherein the effective amount of the CAR T-cells is 104to 109cells per kg body weight of the human patient.

[0272] Embodiment 60. The method of embodiment 59, wherein the effective amount of the CAR T-cells is 105to 106cells per kg body weight of the human patient.

[0273] Embodiment 61. The method of any one of embodiments 1, 3-9, 13, 15, 19, 21, 25, 27, 34, 35-43, and 49-60, wherein ITK inhibitor is administered to the patient before the CAR T- cells are administered to the patient.

[0274] Embodiment 62. The method of any one of embodiments 1, 3-9, 13, 15, 19, 21, 25, 27, 34, 35-43, and 49-60, wherein the ITK inhibitor is administered to the patient after the CAR T- cells are administered to the patient.

[0275] Embodiment 63. The method of any one of embodiments 1, 3-9, 13, 15, 19, 21, 25, 27, 34, 35-43, and 49-60, wherein ITK inhibitor and CAR T-cells are concomitantly administered to the patient.

[0276] Embodiment 64. The method of any one of embodiments 1 to 63, wherein the effective amount of the of the interleukin-2-inducible T-cell kinase inhibitor is about 250 mg to about 1,000 mg per day.

[0277] Embodiment 65. The method of embodiment 64, wherein the effective amount of the of the interleukin-2-inducible T-cell kinase inhibitor is about 300 mg to about 500 mg per day.

[0278] Embodiment 66. The method of embodiment 64, wherein the effective amount of the of the interleukin-2-inducible T-cell kinase inhibitor is about 400 mg per day.

[0279] Embodiment 67. The method of any one of embodiments 1 to 66, wherein the interleukin-2-inducible T-cell kinase inhibitor has a KD or an IC50 ≤ 10 nM to interleukin-2- inducible T-cell kinase.

[0280] Embodiment 68. The method of embodiment 67, wherein the interleukin-2-inducible T-cell kinase inhibitor has a KD or an IC50 ≤ 5 nM to interleukin-2-inducible T-cell kinase.

[0281] Embodiment 69. The method of any one of embodiments 1 to 68, wherein the interleukin-2-inducible T-cell kinase inhibitor has a KD or an IC50 greater than 10 nM to resting lymphocyte kinase.

[0282] Embodiment 70. The method of embodiment 69, wherein the interleukin-2-inducible T-cell kinase inhibitor has a KD or an IC50greater than 100 nM to resting lymphocyte kinase.

[0283] Embodiment 71. The method of embodiment 69, wherein the interleukin-2-inducible T-cell kinase inhibitor has a KD or an IC50greater than 1,000 nM to resting lymphocyte kinase.

[0284] Embodiment 72. The method of any one of embodiments 1 to 66, wherein theinterleukin-2-inducible T-cell kinase inhibitor is not ibrutinib.

[0285] Embodiment 73. The method of any one of embodiments 1 to 66 wherein the interleukin-2-inducible T-cell kinase inhibitor is soquelitinib or a pharmaceutically acceptable salt thereof.

[0286] Embodiment 74. The method of any one of embodiments 1 to 66, wherein the interleukin-2-inducible T-cell kinase inhibitor is a compound of Formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) is: , --NR1ASO2R1D, -NR1AC(O)R1C, -NR1AC(O)OR1C, -NR1AOR1C, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R2is independently hydrogen, halogen, -CX23, -CHX22, -CH2X2, -OCX23, - OCH2X2, -OCHX22, -CN, -SOn2R2D, -SOv2NR2AR2B, -NHC(O)NR2AR2B, -N(O)m2,-NR2AR2B, -C(O)R2C, -C(O)-OR2C, -C(O)NR2AR2B, -OR2D, -NR2ASO2R2D, -NR2AC(O)R2C, -NR2AC(O)OR2C, -NR2AOR2C, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R3is independently hydrogen, halogen, -CX33, -CHX32, -CH2X3, -OCX33, -OCH2X3, -OCHX32, -CN, -SOn3R3D, -SOv3NR3AR3B, -NHC(O)NR3AR3B, -N(O)m3, -NR3AR3B, -C(O)R3C, -C(O)-OR3C, -C(O)NR3AR3B, -OR3D, -NR3ASO2R3D, -NR3AC(O)R3C, -NR3AC(O)OR3C, -NR3AOR3C, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R4is independently hydrogen, halogen, -CX43, -CHX42, -CH2X4, -OCX43, -OCH2X4, -OCHX42, -CN, -SOn4R4D, -SOv4NR4AR4B, -NHC(O)NR4AR4B, -N(O)m4, -NR4AR4B, -C(O)R4C, -C(O)-OR4C, -C(O)NR4AR4B, -OR4D, -NR4ASO2R4D, -NR4AC(O)R4C, -NR4AC(O)OR4C, -NR4AOR4C, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R5is independently substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; L1is -O-, -S-, or substituted or unsubstituted C1-C2alkylene, or substituted or unsubstituted 2 membered heteroalkylene; L2is a bond, -NH-, or -NHC(O)-; L3is a bond, -S(O)2-, -N(R6)-, -O-, -S-, -C(O)-, -C(O)N(R6)-, -N(R6)C(O)-, -N(R6)C(O)NH-, -NHC(O)N(R6)-, -C(O)O-, -OC(O)-, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene; R6is independently hydrogen, -CX63, -CHX62, -CH2X6, -CN, -C(O)R6C, -C(O)OR6C, -C(O)NR6AR6B, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; L4is substituted or unsubstituted heterocycloalkylene; E is: or4C, -CHX2, -CH2X, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R1Aand R1Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R2Aand R2Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R3Aand R3Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R4Aand R4Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R6Aand R6Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R7Aand R7Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; each X, X1, X2, X3, X4, and X6is independently –F, -Cl, -Br, or –I; n1, n2, n3, and n4 are independently an integer from 0 to 2; m1, m2, m3, m4, v1, v2, v3, and v4 are independently 1 to 2; R15is independently hydrogen, halogen, -CX153, -CHX152, -CH2X15, -CN, -SOn15R15D, -SOv15NR15AR15B, -NHNR15AR15B, -ONR15AR15B, -NHC=(O)NHNR15AR15B, -NHC(O)NR15AR15B, -N(O)m15, -NR15AR15B, alkyl,or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; R16is independently hydrogen, halogen, -CX163, -CHX162, -CH2X16, -CN, -SOn16R16D, -SOv16NR16AR16B, -NHNR16AR16B, -ONR16AR16B, -NHC=(O)NHNR16AR16B, -NHC(O)NR16AR16B, -N(O)m16, -NR16AR16B, -C(O)R16C, -C(O)-OR16C, -C(O)NR16AR16B, -OR16D, -NR16ASO2R16D, -NR16AC(O)R16C, -NR16AC(O)OR16C, -NR16AOR16C, -OCX163, -OCHX162, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl, substituted or unsubstituted heteroaryl; R17is independently hydrogen, halogen, -CX173, -CHX172, -CH2X17, -CN, -SOn17R17D, -SOv17NR17AR17B, -NHNR17AR17B, -ONR17AR17B, -NHC=(O)NHNR17AR17B, -NHC(O)NR17AR17B, -N(O)m17, -NR17AR17B, -C(O)R17C, -C(O)-OR17C, -C(O)NR17AR17B, -OR17D, -NR17ASO2R17D, -NR17AC(O)R17C, -NR17AC(O)OR17C, -NR17AOR17C, -OCX173, -OCHX172, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; R18is independently hydrogen, -CX183, -CHX182, -CH2X18, -C(O)R18C, -C(O)OR18C, -C(O)NR18AR18B, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; R15A, R15B, R15C, R15D, R16A, R16B, R16C, R16D, R17A, R17B, R17C, R17D, R18A, R18B, and R18Care each independently hydrogen, -CX3, -CN, -COOH, -CONH2, -CHX2, -CH2X, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R15Aand R15Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R16Aand R16Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R17Aand R17Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R18Aand R18Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; each X, X15, X16, X17and X18is independently –F, -Cl, -Br, or –I; n15, n16, and n17 are independently an integer from 0 to 2; v15, v16, and v17 are independently 1 or 2; and m15, m16, and m17 are independently 1 or 2.

[0287] Embodiment 75. The method of embodiment 72, wherein the interleukin-2-inducible T-cell kinase inhibitor is a compound of Formula (II) or a pharmaceutically acceptable salt thereof, having the formula:I); wherein: R1is hydrogen, haloge OCX13,-OCH2X1, -OCHX12, -CN, -SOn1R1D, -SOv1NR1AR1B, -NHC(O)NR1AR1B, -N(O)m1, -NR1AR1B, -C(O)R1C, -C(O)-OR1C, -C(O)NR1AR1B, -OR1D, -NR1ASO2R1D, -NR1AC(O)R1C, -NR1AC(O)OR1C, -NR1AOR1C, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R3is hydrogen, halogen, -CX33, -CHX32, -CH2X3, -OCX33, -OCH2X3, -OCHX32, -CN, -SOn3R3D, -SOv3NR3AR3B, -NHC(O)NR3AR3B, -N(O)m3, -NR3AR3B, -C(O)R3C, -C(O)-OR3C, -C(O)NR3AR3B, -OR3D, -NR3ASO2R3D, -NR3AC(O)R3C, -NR3AC(O)OR3C, -NR3AOR3C, unsubstituted or substituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R5is unsubstituted or substituted cycloalkyl, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; L3is -C(O)-, a bond, -N(R6)-, or -C(O)N(R6)-; R6is hydrogen or methyl; L4is substituted or unsubstituted 5 to 8 membered monocyclic heterocycloalkylene; E is -C(O)CH=CH2; R1A, R1B, R1C, R1D, R3A, R3B, R3C, and R3Dare each independently hydrogen, -CX3, -CN, -COOH, -CONH2, -CHX2, -CH2X, unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl, or unsubstituted heteroaryl; X, X1, and X3are each independently –F, -Cl, -Br, or –I; n1 and n3 are independently an integer from 0 to 4; and m1, m3, v1, and v3 are independently 1 or 2.

[0288] Embodiment 76. A kit comprising an interleukin-2-inducible T-cell kinase inhibitor and CAR T-cells.

[0289] Embodiment 77. The kit of embodiment 76, wherein the CAR T-cells are CD4+ T- cells.

[0290] Embodiment 78. The kit of embodiment 76, wherein the CAR T-cells are CD8+ T- cells.

[0291] Embodiment 79. The kit of embodiment 76, wherein the CAR T-cells are a combination of CD4+ T-cells and CD8+ T-cells.

[0292] Embodiment 80. The kit of any one of embodiments 76 to 79, wherein the CAR T-cells are autologous.

[0293] Embodiment 81. The kit of any one of embodiments 76 to 79, wherein the CAR T-cells are allogeneic.

[0294] Embodiment 82. The kit of any one of embodiments 76 to 81, wherein the CAR T-cells express a chimeric antigen receptor comprising an extracellular domain that binds to an antigen on cancer cells, wherein the antigen is epidermal growth factor receptor, mesothelin, prostate- specific membrane antigen, disialoganglioside GD2, interleukin-13Ra2, glypican-3, carbonic anhydrase IX, L1CAM, fibroblast activation protein, a cancer testis antigen, MUC1, FOLR1, BCMA, CD133, alpha-fetoprotein, carcinoembryonic antigen, trophoblast glycoproteinprecursor, immature laminin receptor protein, fetal sulphoglycoprotein, g-fetoprotein, cc2-H-ferroportin, beta-2-microglobulin, beta-human chorionic gonadotropin, calcitonin, bladder tumor antigen, CD7, CD19, CD117, CA15-3, CA19-9, CA-125, CA 27.29, CA72-4, CD20, CD22, CD25, CD30, CD33, chromogranin A, cytokeratin fragment 21-1, estrogen receptor, progesterone receptor, fibrinogen, gastrin, HE4, Her2 / neu, neuron-specific enolase, nuclear matrix protein 22, prostatic acid phosphatase, PD-L1, PD-1, prostate-specific antigen, somatostatin receptor, thyroglobulin, 5-HIAA, osteocalcin, transferrin recepto, alkaline phosphtase, BRAF, KRAS, NMP22, BRCA2, urokinase plasminogen activator, apoliprotein A1, S100, nestin, cytokeratin fragments 21-1, ferritin, tissue polypeptide antigen, epididymal secretory protein E4, CECAM 5, CECAM 6, serum M-protein, CTLA-4, B7-1 / B7-2, epithelial tumor antigen, tyrosinase, melanoma-associated antigen, p53, mutated p53, MART-2, beta- catenin, mutated ras, mutated KRAS, BAGE, GAGE-1, NY-ESO-1, PRAME, CT83, SSX2, ART-4, BAGE, beta-catenin / m, Bcr-abL CAMEL, CAP-1 , CASP-8, CDC27 / m, CD 4 / m, a cell surface protein of the claudin family, c-MYC, CT, Cyp-B, DAM, ELF2M, ETV6-AML1, G250, GAGE, GnT-V, KPL-1, HAGE, HPV-E7, HPV-E6, HAST-2, hTERT, LAGE, LDLR / FUT, MAGE- A, MAGE-B, MAGE-C, MART-1, Melan-A, MCIR-1, myosin, MUM-1, MUM-2, MUM-3, NA88-A, NF1, NY-ESO-1, NY-BR-1, HSP90, BCR-ABL-1, RAR, PRAME, proteinase 3, PSM, RAGE, RU1, RU2, SAGE, SART-1, SART-3, SCGB3A2, SCP 1, SCP2, SCP3, SSX, Survrvin, TEL / AML1, TPEM, TRP-1, TRP-2, TRP-2-INT2, TPTE, WT, or a combination of two or more thereof.

[0295] Embodiment 83. The kit of any one of embodiments 76 to 81, wherein the CAR T-cells express a chimeric antigen receptor comprising an extracellular domain that binds to an antigenon cancer cells, wherein the antigen is CD19 or BCMA.

[0296] Embodiment 84. The kit of any one of embodiments 76 to 81, wherein the CAR T-cells express a chimeric antigen receptor comprising an extracellular domain that binds to an antigen on cancer cells, wherein the antigen is epidermal growth factor receptor, mesothelin, prostate- specific membrane antigen, carcinoembryonic antigen, disialoganglioside GD2, interleukin- 13Ra2, glypican-3, carbonic anhydrase IX, L1CAM, CA-125, CD133, fibroblast activation protein, a cancer testis antigen, MUC1, or FOLR1.

[0297] Embodiment 85. The kit of embodiment 84, wherein the epidermal growth factor receptor is epidermal growth factor receptor variant III or human epidermal growth factor receptor 2.

[0298] Embodiment 86. The kit of any one of embodiments 76 to 81, wherein the CAR T-cells are a CAR T-cell therapy selected from the group consisting of axicabtagene ciloleucel,brexucabtagene autoleucel, ciltacabtagene autoleucel, idecabtagene vicleucel, lisocabtagenemaraleucel, and tisagenlecleucel.

[0299] Embodiment 87. The kit of any one of embodiments 76 to 86, wherein the effective amount of the CAR T-cells is 104to 109cells per kg body weight of the human patient.

[0300] Embodiment 88. The kit of embodiment 87, wherein the effective amount of the CAR T-cells is 105to 106cells per kg body weight of the human patient.

[0301] Embodiment 89. The kit of any one of embodiments 76 to 88, wherein the interleukin- 2-inducible T-cell kinase inhibitor has a KD or an IC50 ≤ 10 nM to interleukin-2-inducible T-cell kinase.

[0302] Embodiment 90. The kit of embodiment 89, wherein the interleukin-2-inducible T-cell kinase inhibitor has a KD or an IC50≤ 5 nM to interleukin-2-inducible T-cell kinase.

[0303] Embodiment 91. The kit of any one of embodiments 76 to 90, wherein the interleukin- 2-inducible T-cell kinase inhibitor has a KD or an IC50greater than 10 nM to resting lymphocyte kinase.

[0304] Embodiment 92. The kit of embodiment 91, wherein the interleukin-2-inducible T-cell kinase inhibitor has a KD or an IC50greater than 100 nM to resting lymphocyte kinase.

[0305] Embodiment 93. The kit of embodiment 91, wherein the interleukin-2-inducible T-cell kinase inhibitor has a KD or an IC50greater than 1,000 nM to resting lymphocyte kinase.

[0306] Embodiment 94. The kit of any one of embodiments 76 to 88, wherein the interleukin- 2-inducible T-cell kinase inhibitor is not ibrutinib.

[0307] Embodiment 95. The kit of any one of embodiments 76 to 88, wherein the interleukin-2-inducible T-cell kinase inhibitor is soquelitinib or a pharmaceutically acceptable salt thereof.

[0308] Embodiment 96. The kit of any one of embodiments 76 to 88, wherein the interleukin- 2-inducible T-cell kinase inhibitor is a compound of Formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) is: , ,independently hydrogen, halogen, -CX13, -CHX12, --CN, -SOn1R1D, -SOv1NR1AR1B, -NHC(O)NR1AR1B, -N(O)m1, -NR1AR1B, -C(O)R1C, -C(O)-OR1C, -C(O)NR1AR1B, -OR1D, -NR1ASO2R1D, -NR1AC(O)R1C, -NR1AC(O)OR1C, -NR1AOR1C, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R2is independently hydrogen, halogen, -CX23, -CHX22, -CH2X2, -OCX23, - OCH2X2, -OCHX22, -CN, -SOn2R2D, -SOv2NR2AR2B, -NHC(O)NR2AR2B, -N(O)m2, -NR2AR2B, -C(O)R2C, -C(O)-OR2C, -C(O)NR2AR2B, -OR2D, -NR2ASO2R2D, -NR2AC(O)R2C, -NR2AC(O)OR2C, -NR2AOR2C, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R3is independently hydrogen, halogen, -CX33, -CHX32, -CH2X3, -OCX33, -OCH2X3, -OCHX32, -CN, -SOn3R3D, -SOv3NR3AR3B, -NHC(O)NR3AR3B, -N(O)m3, -NR3AR3B, -C(O)R3C, -C(O)-OR3C, -C(O)NR3AR3B, -OR3D, -NR3ASO2R3D, -NR3AC(O)R3C, -NR3AC(O)OR3C, -NR3AOR3C, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R4is independently hydrogen, halogen, -CX43, -CHX42, -CH2X4, -OCX43, -OCH2X4, -OCHX42, -CN, -SOn4R4D, -SOv4NR4AR4B, -NHC(O)NR4AR4B, -N(O)m4, -NR4AR4B, -C(O)R4C, -C(O)-OR4C, -C(O)NR4AR4B, -OR4D, -NR4ASO2R4D, -NR4AC(O)R4C, -NR4AC(O)OR4C, -NR4AOR4C, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R5is independently substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; L1is -O-, -S-, or substituted or unsubstituted C1-C2 alkylene, or substituted or unsubstituted 2 membered heteroalkylene; L2is a bond, -NH-, or -NHC(O)-; L3is a bond, -S(O)2-, -N(R6)-, -O-, -S-, -C(O)-, -C(O)N(R6)-, -N(R6)C(O)-, -N(R6)C(O)NH-, -NHC(O)N(R6)-, -C(O)O-, -OC(O)-, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene; R6is independently hydrogen, -CX63, -CHX62, -CH2X6, -CN, -C(O)R6C, -C(O)OR6C, -C(O)NR6AR6B, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; L4is substituted or unsubstituted heterocycloalkylene; E is:R4D, R6A, R6B, and R6Care each independently hydrogen, -CX3, -CN, -COOH, -CONH2, -CHX2, -CH2X, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R1Aand R1Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R2Aand R2Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R3Aand R3Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R4Aand R4Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R6Aand R6Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R7Aand R7Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; each X, X1, X2, X3, X4, and X6is independently –F, -Cl, -Br, or –I; n1, n2, n3, and n4 are independently an integer from 0 to 2; m1, m2, m3, m4, v1, v2, v3, and v4 are independently 1 to 2; R15is independently hydrogen, halogen, -CX153, -CHX152, -CH2X15, -CN, -SOn15R15D, -SOv15NR15AR15B, -NHNR15AR15B, -ONR15AR15B, -NHC=(O)NHNR15AR15B, -NHC(O)NR15AR15B, -N(O)m15, -NR15AR15B, -C(O)R15C, -C(O)-OR15C, -C(O)NR15AR15B, -OR15D, -NR15ASO2R15D, -NR15AC(O)R15C, -NR15AC(O)OR15C, -NR15AOR15C, -OCX153, -OCHX152, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; R16is independently hydrogen, halogen, -CX163, -CHX162, -CH2X16, -CN, -SOn16R16D, -SOv16NR16AR16B, -NHNR16AR16B, -ONR16AR16B, -NHC=(O)NHNR16AR16B, -NHC(O)NR16AR16B, -N(O)m16, -NR16AR16B, -C(O)R16C, -C(O)-OR16C, -C(O)NR16AR16B, -OR16D, -NR16ASO2R16D, -NR16AC(O)R16C, -NR16AC(O)OR16C, -NR16AOR16C, -OCX163, -OCHX162, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; R17is independently hydrogen, halogen, -CX173, -CHX172, -CH2X17, -CN, -SOn17R17D, -SOv17NR17AR17B, -NHNR17AR17B, -ONR17AR17B, -NHC=(O)NHNR17AR17B, -NHC(O)NR17AR17B, -N(O)m17, -NR17AR17B, -C(O)R17C, -C(O)-OR17C, -C(O)NR17AR17B, -OR17D, -NR17ASO2R17D, -NR17AC(O)R17C,-NR17AC(O)OR17C, -NR17AOR17C, -OCX173, -OCHX172, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; R18is independently hydrogen, -CX183, -CHX182, -CH2X18, -C(O)R18C, -C(O)OR18C, -C(O)NR18AR18B, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; R15A, R15B, R15C, R15D, R16A, R16B, R16C, R16D, R17A, R17B, R17C, R17D, R18A, R18B, and R18Care each independently hydrogen, -CX3, -CN, -COOH, -CONH2, -CHX2, -CH2X, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R15Aand R15Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R16Aand R16Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R17Aand R17Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R18Aand R18Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; each X, X15, X16, X17and X18is independently –F, -Cl, -Br, or –I; n15, n16, and n17 are independently an integer from 0 to 2; v15, v16, and v17 are independently 1 or 2; and m15, m16, and m17 are independently 1 or 2.

[0309] Embodiment 97. The kit of embodiment 96, wherein the interleukin-2-inducible T-cell kinase inhibitor is a compound of Formula (II) or a pharmaceutically acceptable salt thereof, having the formula: ; wherein: R1is hydrogen,OCH2X1, -OCHX12, -CN, -SOn1R1D, -SOv1NR1AR1B, -NHC(O)NR1AR1B, -N(O)m1, -NR1AR1B, -C(O)R1C,-C(O)-OR1C, -C(O)NR1AR1B, -OR1D, -NR1ASO2R1D, -NR1AC(O)R1C, -NR1AC(O)OR1C, -NR1AOR1C, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R3is hydrogen, halogen, -CX33, -CHX32, -CH2X3, -OCX33, -OCH2X3, -OCHX32, -CN, -SOn3R3D, -SOv3NR3AR3B, -NHC(O)NR3AR3B, -N(O)m3, -NR3AR3B, -C(O)R3C, -C(O)-OR3C, -C(O)NR3AR3B, -OR3D, -NR3ASO2R3D, -NR3AC(O)R3C, -NR3AC(O)OR3C, -NR3AOR3C, unsubstituted or substituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R5is unsubstituted or substituted cycloalkyl, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; L3is -C(O)-, a bond, -N(R6)-, or -C(O)N(R6)-; R6is hydrogen or methyl; L4is substituted or unsubstituted 5 to 8 membered monocyclic heterocycloalkylene; E is -C(O)CH=CH2; R1A, R1B, R1C, R1D, R3A, R3B, R3C, and R3Dare each independently hydrogen, -CX3, -CN, -COOH, -CONH2, -CHX2, -CH2X, unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl, or unsubstituted heteroaryl; X, X1, and X3are each independently –F, -Cl, -Br, or –I; n1 and n3 are independently an integer from 0 to 4; and m1, m3, v1, and v3 are independently 1 or 2.

[0310] Embodiment 98. A composition comprising an interleukin-2-inducible T-cell kinase inhibitor and CAR T-cells.

[0311] Embodiment 99. The composition of embodiment 98, wherein the CAR T-cells are CD4+ T-cells.

[0312] Embodiment 100. The composition of embodiment 98, wherein the CAR T-cells are CD8+ T-cells.

[0313] Embodiment 101. The composition of embodiment 98, wherein the CAR T-cells are a combination of CD4+ T-cells and CD8+ T-cells.

[0314] Embodiment 102. The composition of any one of embodiments 98 to 101, wherein the CAR T-cells are autologous.

[0315] Embodiment 103. The composition of any one of embodiments 98 to 101, wherein the CAR T-cells are allogeneic.

[0316] Embodiment 104. The composition of any one of embodiments 98 to 103, wherein the CAR T-cells express a chimeric antigen receptor comprising an extracellular domain that bindsto an antigen on cancer cells, wherein the antigen is epidermal growth factor receptor, mesothelin, prostate-specific membrane antigen, disialoganglioside GD2, interleukin-13Ra2, glypican-3, carbonic anhydrase IX, L1CAM, fibroblast activation protein, a cancer testis antigen, MUC1, FOLR1, BCMA, CD133, alpha-fetoprotein, carcinoembryonic antigen,trophoblast glycoprotein precursor, immature laminin receptor protein, fetal sulphoglycoprotein,g-fetoprotein, cc2-H-ferroportin, beta-2-microglobulin, beta-human chorionic gonadotropin, calcitonin, bladder tumor antigen, CD7, CD19, CD117, CA15-3, CA19-9, CA-125, CA 27.29, CA72-4, CD20, CD22, CD25, CD30, CD33, chromogranin A, cytokeratin fragment 21-1, estrogen receptor, progesterone receptor, fibrinogen, gastrin, HE4, Her2 / neu, neuron-specific enolase, nuclear matrix protein 22, prostatic acid phosphatase, PD-L1, PD-1, prostate-specific antigen, somatostatin receptor, thyroglobulin, 5-HIAA, osteocalcin, transferrin recepto, alkaline phosphtase, BRAF, KRAS, NMP22, BRCA2, urokinase plasminogen activator, apoliprotein A1, S100, nestin, cytokeratin fragments 21-1, ferritin, tissue polypeptide antigen, epididymal secretory protein E4, CECAM 5, CECAM 6, serum M-protein, CTLA-4, B7-1 / B7-2, epithelial tumor antigen, tyrosinase, melanoma-associated antigen, p53, mutated p53, MART-2, beta- catenin, mutated ras, mutated KRAS, BAGE, GAGE-1, NY-ESO-1, PRAME, CT83, SSX2, ART-4, BAGE, beta-catenin / m, Bcr-abL CAMEL, CAP-1 , CASP-8, CDC27 / m, CD 4 / m, a cell surface protein of the claudin family, c-MYC, CT, Cyp-B, DAM, ELF2M, ETV6-AML1, G250, GAGE, GnT-V, KPL-1, HAGE, HPV-E7, HPV-E6, HAST-2, hTERT, LAGE, LDLR / FUT, MAGE- A, MAGE-B, MAGE-C, MART-1, Melan-A, MCIR-1, myosin, MUM-1, MUM-2, MUM-3, NA88-A, NF1, NY-ESO-1, NY-BR-1, HSP90, BCR-ABL-1, RAR, PRAME, proteinase 3, PSM, RAGE, RU1, RU2, SAGE, SART-1, SART-3, SCGB3A2, SCP 1, SCP2, SCP3, SSX, Survrvin, TEL / AML1, TPEM, TRP-1, TRP-2, TRP-2-INT2, TPTE, WT, or a combination of two or more thereof.

[0317] Embodiment 105. The composition of any one of embodiments 98 to 103, wherein the CAR T-cells express a chimeric antigen receptor comprising an extracellular domain that binds to an antigen on cancer cells, wherein the antigen is CD19 or BCMA.

[0318] Embodiment 106. The composition of any one of embodiments 98 to 103, wherein the CAR T-cells express a chimeric antigen receptor comprising an extracellular domain that binds to an antigen on cancer cells, wherein the antigen is epidermal growth factor receptor, mesothelin, prostate-specific membrane antigen, carcinoembryonic antigen, disialoganglioside GD2, interleukin-13Ra2, glypican-3, carbonic anhydrase IX, L1CAM, CA-125, CD133, fibroblast activation protein, a cancer testis antigen, MUC1, or FOLR1.

[0319] Embodiment 107. The composition of embodiment 106, wherein the epidermal growthfactor receptor is epidermal growth factor receptor variant III or human epidermal growth factor receptor 2.

[0320] Embodiment 108. The composition of any one of embodiments 98 to 103, wherein the CAR T-cells are a CAR T-cell therapy selected from the group consisting of axicabtageneciloleucel, brexucabtagene autoleucel, ciltacabtagene autoleucel, idecabtagene vicleucel,lisocabtagene maraleucel, and tisagenlecleucel.

[0321] Embodiment 109. The composition of any one of embodiments 98 to 108, wherein the effective amount of the CAR T-cells is 104to 109cells per kg body weight of the human patient.

[0322] Embodiment 110. The composition of embodiment 109, wherein the effective amount of the CAR T-cells is 105to 106cells per kg body weight of the human patient.

[0323] Embodiment 111. The composition of any one of embodiments 98 to 110, wherein the interleukin-2-inducible T-cell kinase inhibitor has a KD or an IC50≤ 10 nM to interleukin-2- inducible T-cell kinase.

[0324] Embodiment 112. The composition of embodiment 111, wherein the interleukin-2- inducible T-cell kinase inhibitor has a KD or an IC50 ≤ 5 nM to interleukin-2-inducible T-cell kinase.

[0325] Embodiment 113. The composition of any one of embodiments 98 to 112, wherein the interleukin-2-inducible T-cell kinase inhibitor has a KD or an IC50 greater than 10 nM to resting lymphocyte kinase.

[0326] Embodiment 114. The composition of embodiment 113, wherein the interleukin-2- inducible T-cell kinase inhibitor has a KD or an IC50greater than 100 nM to resting lymphocyte kinase.

[0327] Embodiment 115. The composition of embodiment 113, wherein the interleukin-2- inducible T-cell kinase inhibitor has a KD or an IC50greater than 1,000 nM to resting lymphocyte kinase.

[0328] Embodiment 116. The composition of any one of embodiments 98 to 110, wherein the interleukin-2-inducible T-cell kinase inhibitor is not ibrutinib.

[0329] Embodiment 117. The composition of any one of embodiments 98 to 110, wherein the interleukin-2-inducible T-cell kinase inhibitor is soquelitinib or a pharmaceutically acceptable salt thereof.

[0330] Embodiment 118. The composition of any one of embodiments 98 to 110, wherein the interleukin-2-inducible T-cell kinase inhibitor is a compound of Formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) is:, 2, - D, -NR1ASO2R1D, -NR1AC(O)R1C, -NR1AC(O)OR1C, -NR1AOR1C, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R2is independently hydrogen, halogen, -CX23, -CHX22, -CH2X2, -OCX23, - OCH2X2, -OCHX22, -CN, -SOn2R2D, -SOv2NR2AR2B, -NHC(O)NR2AR2B, -N(O)m2, -NR2AR2B, -C(O)R2C, -C(O)-OR2C, -C(O)NR2AR2B, -OR2D, -NR2ASO2R2D, -NR2AC(O)R2C, -NR2AC(O)OR2C, -NR2AOR2C, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R3is independently hydrogen, halogen, -CX33, -CHX32, -CH2X3, -OCX33, -OCH2X3, -OCHX32, -CN, -SOn3R3D, -SOv3NR3AR3B, -NHC(O)NR3AR3B, -N(O)m3, -NR3AR3B, -C(O)R3C, -C(O)-OR3C, -C(O)NR3AR3B, -OR3D, -NR3ASO2R3D, -NR3AC(O)R3C, -NR3AC(O)OR3C, -NR3AOR3C,substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R4is independently hydrogen, halogen, -CX43, -CHX42, -CH2X4, -OCX43, -OCH2X4, -OCHX42, -CN, -SOn4R4D, -SOv4NR4AR4B, -NHC(O)NR4AR4B, -N(O)m4, -NR4AR4B, -C(O)R4C, -C(O)-OR4C, -C(O)NR4AR4B, -OR4D, -NR4ASO2R4D, -NR4AC(O)R4C, -NR4AC(O)OR4C, -NR4AOR4C, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R5is independently substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; L1is -O-, -S-, or substituted or unsubstituted C1-C2 alkylene, or substituted or unsubstituted 2 membered heteroalkylene; L2is a bond, -NH-, or -NHC(O)-; L3is a bond, -S(O)2-, -N(R6)-, -O-, -S-, -C(O)-, -C(O)N(R6)-, -N(R6)C(O)-, -N(R6)C(O)NH-, -NHC(O)N(R6)-, -C(O)O-, -OC(O)-, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene; R6is independently hydrogen, -CX63, -CHX62, -CH2X6, -CN, -C(O)R6C, -C(O)OR6C, -C(O)NR6AR6B, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; L4is substituted or unsubstituted heterocycloalkylene; E is:-CHX2, -CH2X, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R1Aand R1Bsubstituents bondedto the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R2Aand R2Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R3Aand R3Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R4Aand R4Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R6Aand R6Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R7Aand R7Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; each X, X1, X2, X3, X4, and X6is independently –F, -Cl, -Br, or –I; n1, n2, n3, and n4 are independently an integer from 0 to 2; m1, m2, m3, m4, v1, v2, v3, and v4 are independently 1 to 2; R15is independently hydrogen, halogen, -CX153, -CHX152, -CH2X15, -CN, -SOn15R15D, -SOv15NR15AR15B, -NHNR15AR15B, -ONR15AR15B, -NHC=(O)NHNR15AR15B, -NHC(O)NR15AR15B, -N(O)m15, -NR15AR15B, -C(O)R15C, -C(O)-OR15C, -C(O)NR15AR15B, -OR15D, -NR15ASO2R15D, -NR15AC(O)R15C, -NR15AC(O)OR15C, -NR15AOR15C, -OCX153, -OCHX152, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; R16is independently hydrogen, halogen, -CX163, -CHX162, -CH2X16, -CN, -SOn16R16D, -SOv16NR16AR16B, -NHNR16AR16B, -ONR16AR16B, -NHC=(O)NHNR16AR16B, -NHC(O)NR16AR16B, -N(O)m16, -NR16AR16B, -C(O)R16C, -C(O)-OR16C, -C(O)NR16AR16B, -OR16D, -NR16ASO2R16D, -NR16AC(O)R16C, -NR16AC(O)OR16C, -NR16AOR16C, -OCX163, -OCHX162, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; R17is independently hydrogen, halogen, -CX173, -CHX172, -CH2X17, -CN, -SOn17R17D, -SOv17NR17AR17B, -NHNR17AR17B, -ONR17AR17B, -NHC=(O)NHNR17AR17B, -NHC(O)NR17AR17B, -N(O)m17, -NR17AR17B, -C(O)R17C, -C(O)-OR17C, -C(O)NR17AR17B, -OR17D, -NR17ASO2R17D, -NR17AC(O)R17C, -NR17AC(O)OR17C, -NR17AOR17C, -OCX173, -OCHX172, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; R18is independently hydrogen, -CX183, -CHX182, -CH2X18, -C(O)R18C, -C(O)OR18C,-C(O)NR18AR18B, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; R15A, R15B, R15C, R15D, R16A, R16B, R16C, R16D, R17A, R17B, R17C, R17D, R18A, R18B, and R18Care each independently hydrogen, -CX3, - - - - - substituted or unsubstituted substituted oror unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R15Aand R15Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R16Aand R16Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R17Aand R17Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; R18Aand R18Bsubstituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; each X, X15, X16, X17and X18is independently –F, -Cl, -Br, or –I; n15, n16, and n17 are independently an integer from 0 to 2; v15, v16, and v17 are independently 1 or 2; and m15, m16, and m17 are independently 1 or 2.

[0331] Embodiment 119. The composition of embodiment 118, wherein the interleukin-2- inducible T-cell kinase inhibitor is a compound of Formula (II) or a pharmaceutically acceptable salt thereof, having the formula: ; wherein: R1is hydrogen,OCH2X1, -OCHX12, -CN, -SOn1R1D, -SOv1NR1AR1B, -NHC(O)NR1AR1B, -N(O)m1, -NR1AR1B, -C(O)R1C, -C(O)-OR1C, -C(O)NR1AR1B, -OR1D, -NR1ASO2R1D, -NR1AC(O)R1C, -NR1AC(O)OR1C, -NR1AOR1C, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R3is hydrogen, halogen, -CX33,-CHX32, -CH2X3, -OCX33, -OCH2X3, -OCHX32, -CN, -SOn3R3D, -SOv3NR3AR3B, -NHC(O)NR3AR3B, -N(O)m3, -NR3AR3B, -C(O)R3C, -C(O)-OR3C, -C(O)NR3AR3B, -OR3D, -NR3ASO2R3D, -NR3AC(O)R3C, -NR3AC(O)OR3C, -NR3AOR3C, unsubstituted or substituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R5is unsubstituted or substituted cycloalkyl, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; L3is -C(O)-, a bond, -N(R6)-, or -C(O)N(R6)-; R6is hydrogen or methyl; L4is substituted or unsubstituted 5 to 8 membered monocyclic heterocycloalkylene; E is -C(O)CH=CH2; R1A, R1B, R1C, R1D, R3A, R3B, R3C, and R3Dare each independently hydrogen, -CX3, -CN, -COOH, -CONH2, -CHX2, -CH2X, unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl, or unsubstituted heteroaryl; X, X1, and X3are each independently –F, -Cl, -Br, or –I; n1 and n3 are independently an integer from 0 to 4; and m1, m3, v1, and v3 are independently 1 or 2.

[0332] Embodiment 120. The composition of any one of embodiments 98 to 119, wherein the composition further comprises human serum or fetal bovine serum.

[0333] Embodiment 121. The composition of any one of embodiments 98 to 119, wherein the composition further comprises a serum-free medium. EXAMPLES

[0334] The following example is for purposes of illustration only, and is not intended to limit the spirit or scope of the disclosure or claims.

[0335] ITK is a member of the TEC family of kinases that plays a major role in T cell activation by integrating both TCR and CD28 costimulatory signaling pathways. Studies from ITK knockout mice and in vitro studies using shRNA knock-down demonstrate that ITK modulates the magnitude of TCR signaling strength, leading to distinct fates during T cell differentiation and to the activation of distinct subsets of effector functions. This modulatory role for ITK is partially redundant since the structurally similar TEC kinase, RLK, has overlapping functions in Th1 cells. In this report, we describe a covalent inhibitor that is selective for ITK while sparing RLK. Our studies confirm that pharmacologic blockade of ITK by soquelitinib results in the expected inhibition of downstream signaling proteins including pPLCγ1, pERK, pS6, and NFκB and reduced secretion of IL-2. Moreover, inhibition of GATA- 3 expression also was observed, supporting the known intricate relationship between ITK andGATA-3 function and the crucial role of ITK and GATA-3 in Th2 function. Consistent with previously reported ITK knockout studies, we demonstrate preferential inhibition of Th2-derived cytokines such as IL-4, IL-5 and IL-13 with relative sparing of the Th1-derived cytokine, IFNγ. We find that at very high concentrations (10µM), the anti-proliferative effect of soquelitinib dominates over the effects on T cell differentiation probably as a result of inhibition of T cell receptor signaling pathways. Collectively, the selectivity and differential effects of soquelitinib on Th1 versus Th2 cells support the utility as an immunotherapy for cancer by blocking Th2 function without interfering with Th1.

[0336] Soquelitinib inhibited tumor growth in several murine models including tumors that do not express ITK. In a monotherapy setting, we show increased infiltration of CD8+ T cells in both EL4 and CT26 tumor models. Furthermore, tumor-infiltrating T cells possess increased cytolytic capacity as evidenced by elevated production of IFNγ, TNF, and perforin. Combining soquelitinib with immune checkpoint blockade further enhances anti-tumor efficacy, leading to deep tumor regression after treatment was terminated. Several possibilities can explain this enhanced immune response to cancer in the presence of the ITK inhibitor. First, soquelitinib preferentially inhibits Th2 cytokine synthesis. Th2 cytokines have been implicated as negative regulators of the immune response to cancer. There is evidence that targeting the Th2 pathway might alleviate immunosuppression in the tumor microenvironment (TME) by inhibiting the differentiation of M2-like tumor infiltrating macrophages and Th2-promoting DC subset. In addition to relieving immunosuppression, selective blockade of ITK appears to modulate intratumoral balance between Th1 and Th2 cells within the TME. We show that CD4+ tumor infiltrating lymphocytes (TILs) from soquelitinib -treated tumors produce a higher percentage ofcells expressing IFNγ and TNF, dominant cytokines produced by Th1 cells. A previous reporthas shown that the Th1 cytokine, IFNγ can induce a positive feedback loop for its own production by Th1 CD4 T cells. This could potentially reinforce a bias away from a Th2- to Th1-favoring TME, resulting in further inhibition of tumor growth.

[0337] Another piece of evidence to support Th1-favoring TME was that surface expression of CXCR3, a Th1-chemokine receptor, was significantly upregulated in both CD4+ and CD8+ T cells from soquelitinib treated animals. Our result is consistent with the study from CXCR3- deficient mice showing that CXCR3-mediated T cell recruitment to the TME is strongly associated with the induction of Th1 and cytolytic CD8 T cells. Since CXCR3 upregulation is linked to a higher migratory capacity of T cells, this explains increased infiltration of CD8 T cells found in soquelitinib treated tumors.

[0338] Second, selectivity of soquelitinib to inhibit ITK over RLK enables specific blockadeof ITK-mediated immune modulation, while preserving CD8 and Th1 cell functions. In support of this, additional data using a non-selective analog of soquelitinib that reacts with both ITK and RLK, indicate that treatment with an inhibitor that blocks RLK, results in marked reduction in IFNγ, IL-2 and granzyme B production in CD8 T cells (data not shown). This result mirrors the findings from the ITK-and RLK -double knockout mice in which CD8 and Th1 T cell functions were greatly impaired. Together, achieving significant selectivity over RLK avoids impacts on vital T cell functions which are critical in driving anti-tumor immunity.

[0339] Third, preventing or reverting T cell exhaustion after soquelitinib treatment enhances anti-tumor immunity and increases the duration of an antitumor response. It is known that continuous antigen exposure can lead to T cell exhaustion, which is characterized by gradual deterioration of CD8 T cell function and elevated expression of inhibitory receptors such as PD1, TIGIT, LAG3, and TIM3. CD8 exhaustion has become a major barrier to cancer immunotherapy as the dysfunctional state has inverse correlation with clinical prognosis. We find a dose-dependent reversal of the exhaustion process as evidenced by increased production of granzyme B and IFNγ in the presence of up to 1 µM concentrations of soquelitinib (FIGS. 1D-1F). Soquelitinib inhibits acquisition of the epigenetic hallmarks of T cell exhaustion and induces transcriptomic and epigenetic reprograming. Together, our findings show novel utility of soquelitinib in mitigating T cell exhaustion, thereby prolonging anti-tumor response.

[0340] These studies have important implications for tumor immunotherapy. Checkpoint blockade has emerged as a major therapeutic advance for cancer. However, several resistance mechanisms have been elucidated including insufficient tumor immunogenicity, an immunosuppressive TME, defects in IFN^ signaling and T cell exhaustion. ITK blockade can overcome many of these resistance mechanisms and enhance immune checkpoint therapy. In fact, our murine studies demonstrate enhancement of anti-tumor activity when soquelitinib is added to anti-PD1 and anti-CTLA4.

[0341] In conclusion, we have described inhibition of ITK as a novel approach to enhance the anti-tumor immune response, which occurs through several mechanisms including increased infiltration and function of cytotoxic lymphocytes and the reduction and reversal of T cell exhaustion. These observations support the use of a selective ITK inhibitor as a therapy for cancer.

[0342] Experimental Results

[0343] Chemical Design and Synthesis

[0344] Analysis of ligand bound ITK crystal structures in the Protein Data Bank (PDB)suggested the aminothiazole-based molecular scaffold (PDB entry 3MJ2) as a suitable starting point for structure-based drug design of ITK-selective covalent inhibitors. This ligand forms two hydrogen bonds with the Met-438 residue in the hinge region and a hydrophobic interaction with the gatekeeper residue Phe-435. A piperazine-acetamide fragment projects toward Cys-442, providing a useful vector for ligand modification with electrophilic substituents to engage Cys- 442 in covalent bond formation. The benzylamine moiety was replaced with the less bulky and less polar cyclopropyl-substituent. Further optimization of ITK selectivity and potency was achieved with the introduction of the 7-membered 1,4-diazepane ring with the R-methyl group. Presumably, the chiral methyl acts as an anchor to stabilize one of the conformations of the 7- membered ring that directs the acrylamide fragment towards Cys-442 facilitating irreversible covalent bond formation between soquelitinib and ITK via a Michael addition reaction. The selective binding of soquelitinib (M.W.= 514.66) is shown in FIG.3.

[0345] Covalent irreversible inhibition of ITK by soquelitinib

[0346] A chymotryptic digest of soquelitinib -inhibited ITK was analyzed by liquid chromatography with tandem mass spectroscopy (LC MS / MS) to detect the presence of the covalently linked inhibitor to the enzyme. The digest generated 247 peptide fragments and their sequences were identified through deconvolution of the mass spectra. A set of three nested peptides were detected in the sample treated with soquelitinib that were absent from the untreated control sample. Each of the three peptides contained Cys-442 and their masses were shifted by the mass of soquelitinib when compared to the corresponding unmodified peptide sequence. The presence of the three unique peptide fragments generated by soquelitinib treatment is consistent with the inhibitor covalently labeling Cys-442. There were no other peptide fragments detected that were modified by soquelitinib. Collectively these data demonstrate that soquelitinib is a selective, irreversible, and covalent inhibitor of ITK.

[0347] Kinase selectivity of soquelitinib

[0348] The five members of the TEC kinase family and six other enzymes of the human kinome contain a cysteine in a position homologous to Cys-442 in ITK and could potentially be irreversibly inhibited by soquelitinib. To investigate the selectivity against the 11 cysteine- containing kinases, Kd values for soquelitinib for each enzyme were obtained. Soquelitinib displayed aKd = 6.5 nM for ITK and was at least 80-fold selective over the remaining cysteine- containing kinases. Profiling data for ibrutinib, a covalent BTK inhibitor, approved for treatment of B cell lymphomas also is shown for comparison. Soquelitinib was then profiled against the human kinome at the single concentration of 1.0 µM. Soquelitinib displayed a high degree of selectivity such that only eight kinases were inhibited ≥65% and only the targeted kinase, ITK,was inhibited by >95%. We measured kinact / Kivalues for the two kinases, ITK and RLK, and found soquelitinib is 115-fold selective toward ITK over RLK. Moreover, other kinases in the T cell receptor pathway were not inhibited by soquelitinib (kinase activity, percent control: Lck, 89; Fyn, 99; ZAP-70, 59; RLK, 70). See FIG.3.

[0349] Soquelitinib inhibits TCR signaling downstream of ITK and blocks IL-2 production

[0350] To investigate whether inhibition of ITK activity by soquelitinib modulates TCR signaling during activation, we measured phosphorylation of several proteins downstream of TCR signaling such as ZAP70, PLCγ1 in human H9 cells, and ERK and ribosomal protein S6 in human CD4+ T cells, respectively. Soquelitinib inhibited the phosphorylation of PLCγ1, ERK and S6 protein and had a minimal impact on phosphorylation of a proximal kinase, ZAP70. Previous studies have shown that TCR-medicated activation of NF-κB and GATA-3 is ITK dependent. We tested whether blockade of ITK kinase activity by soquelitinib could affect GATA-3 expression which plays an essential role in Th2 cell differentiation. Consistent with the published report, we observed impaired NF-κB activation after soquelitinib treatment for 60 minutes in the T8ML-1 cell line, a cell line derived from a patient with refractory peripheral T cell lymphoma (PTCL) as shown by reduced NF-κB protein levels in both cytoplasmic and nuclear fractions. We found that soquelitinib treatment lowered the expression of both ITK and GATA-3. Together, these results indicated that inhibition of ITK kinase activity by soquelitinib led to inhibition of downstream TCR signaling events and GATA-3 expression. We next investigated the effect on IL-2 secretion, an early event in T cell activation. Soquelitinib suppressed IL-2 secretion in Jurkat cells in response to TCR stimulation with a mean IC50 of 136 nM. Given that IL-2 plays an essential role in T cell proliferation and survival, we evaluated the viability and proliferation of human T cells treated with a range of soquelitinib doses. Our results revealed that at 10µM concentrations of soquelitinib, cell viability was unaffected but total cell number was reduced, findings consistent with an antiproliferative effect seen only at the highest concentration. Viability and proliferation were not affected at lower concentrations of drug.

[0351] Soquelitinib suppresses Th2 associated cytokines and induces Th1 skewing in CD4 Tcells

[0352] We evaluated the effect of soquelitinib on T effector cytokines following 3 days of TCR stimulation in purified CD4 T cells from either healthy donors or Sezary cells from patients with cutaneous T cell lymphomas (CTCL), which highly express GATA-3 and have a Th2 phenotype. When normal human T cells are incubated with increasing concentrations of soquelitinib, Th2 cytokine production (IL-4, IL-5 and IL-13) is reduced. The prototypical Th1cytokine, IFNγ, is affected but only at the highest concentrations of soquelitinib which were tested. Interestingly, Sezary cells exhibited greater sensitivity to soquelitinib as soquelitinib inhibited production of Th2 associated cytokines to a greater extent in Sezary cells than in normal CD4 T cells. Again, their production of IFNγ can also be reduced, but only at the highest concentration of soquelitinib.

[0353] We further validated the effect of soquelitinib on Th1 skewing of naïve CD4 T cells by enumerating the intracellular ratio of IFNγ to IL-4. Naive human CD4 T cells were activated in the presence of varying concentrations of soquelitinib for 6 days. Soquelitinib had a minimal effect on the ratio of IFN^+ / CD4+ T cells at doses up to 1 µM. In contrast, the percentage of IL- 4+CD4+ T cells was significantly reduced at 0.3 and 1 µM. As a result, at 1 µM concentration of soquelitinib, the ratio of IFN^ to IL-4 CD4+ T cells was increased by at least 2-fold. At a concentration of 10 µM, soquelitinib completely abolished production of either IL-4 or IFN^ in naïve CD4+ T cells, possibly due to its antiproliferative effects at high concentrations. Together, our results indicated that soquelitinib induced a Th1-skewing phenotype in naïve CD4 T cells in a non-polarizing culture condition.

[0354] Next, we evaluated the effect of ITK inhibition on already polarized CD4 helper cells. Naïve CD4 cells were stimulated and differentiated under Th1 or Th2 polarizing conditions for 6 days before soquelitinib was introduced into the culture conditions for an additional three days. Similar to ITK inhibition in naïve CD4 T cells, we found that soquelitinib inhibited the synthesis of Th2-associated cytokines such as IL-4, IL-5 and IL-13 in already polarized Th2 cells, while IFN^ production by Th1 was minimally affected. Collectively, our results indicated that ITK inhibition by soquelitinib can skew the balance between Th1 and Th2 cells.

[0355] Safety study.

[0356] The safety of soquelitinib was evaluated in rats given 28 daily oral doses and revealed a no adverse effect level exceeding 1000 mg / kg.

[0357] Soquelitinib shows efficacy as single agent in several murine tumor models. Soquelitinib is orally bioavailable and exhibits sustained ITK target occupancy in vivo. These data prompted us to test the potential of soquelitinib as an immunotherapy for cancer. We first investigated the activity of soquelitinib on tumor growth on a range of murine syngeneic tumor models. These tumor models included CT26 colon adenocarcinoma, RENCA renal cell carcinoma, B16F10 OVA melanoma, A20 B cell lymphoma and EL4, an ITK expressing T cell lymphoma. Administration of soquelitinib as single agent at 30 mg / kg in solution formulation or 130 mg / kg in chow formulation (A20 model) resulted in a significant inhibition of tumor growthin all five tumor models.

[0358] CD8 T cell depletion increases tumor growth in soquelitinib-treated mice

[0359] We then investigated the contribution of cell types to the antitumor effect of CPI-818 by in vivo antibody depletion of CD4, CD8 or NK cells in the CT26 tumor model. As...

Claims

CLAIMS What is claimed is:

1. A method of treating cancer in a patient in need thereof, the method comprising (a) administering to the patient: (i) an effective amount of an interleukin-2- inducible T-cell kinase inhibitor and (ii) an effective amount of CAR T- cells, thereby treating cancer in the patient; or (b) contacting CAR T-cells with an effective amount of an interleukin-2- inducible T-cell kinase inhibitor in vitro, thereby producing ITK- contacted CAR T-cells; and administering to the patient an effective amount of the ITK-contacted CAR T-cells, thereby treating cancer in the patient.

2. The method of claim 1, further comprising administering to the patient an effective amount of the interleukin-2-inducible T-cell kinase inhibitor.

3. The method of claim 1, wherein the cancer is lymphoma, leukemia, lung cancer, colorectal cancer, pancreatic cancer, prostate cancer, breast cancer, gastric cancer, renal cancer, melanoma, or head and neck cancer.

4. The method of claim 1, wherein the cancer is B-cell precursoracute lymphoblastic leukemia, mantle cell lymphoma, follicular lymphoma, or multiplemyeloma.

5. The method of claim 1, wherein the cancer is lymphoma.

6. The method of claim 5, wherein the lymphoma is T-cell lymphoma or B-cell lymphoma.

7. The method of claim 6, wherein the B-cell lymphoma is diffuse large B-cell lymphoma or high grade B-cell lymphoma.

8. A method for preventing and / or reversing T-cell exhaustion in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2-inducible T-cell kinase inhibitor; thereby preventing and / or reversing T-cell exhaustion in the patient.

9. A method for preventing and / or reversing CAR T-cell exhaustion in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2-inducible T-cell kinase inhibitor; wherein the patient is receiving or has receivedCAR T-cell therapy; thereby preventing and / or reversing CAR T-cell exhaustion in the patient.

10. A method of preventing and / or reversing CAR T-cell exhaustion in a patient in need thereof, the method comprising administering to the patient: (i) an effective amount of an interleukin-2-inducible T-cell kinase inhibitor and (ii) an effective amount of CAR T-cells, thereby preventing and / or reducing CAR T-cell exhaustion in the patient.

11. A method of preventing and / or reversing CAR T-cell exhaustion in a patient in need thereof, the method comprising: (i) contacting CAR T-cells with an effective amount of an interleukin-2-inducible T-cell kinase inhibitor in vitro, thereby producing ITK-contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK-contacted CAR T- cells; thereby preventing and / or reducing CAR T-cell exhaustion in the patient.

12. A method for stimulating a T-cell-mediated immune response in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2-inducible T-cell kinase inhibitor; thereby stimulating the T-cell-mediated immune response in the patient.

13. A method for stimulating a CAR T-cell-mediated immune response in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2-inducible T-cell kinase inhibitor; wherein the patient is receiving or has received CAR T-cell therapy; thereby stimulating the CAR T-cell-mediated immune response in the patient.

14. A method for stimulating a CAR T-cell-mediated immune response in a patient in need thereof, the method comprising administering to the patient: (i) an effective amount of an interleukin-2-inducible T-cell kinase inhibitor and (ii) an effective amount of CAR T-cells, thereby stimulating the CAR T-cell-mediated immune response in the patient.

15. A method for stimulating a CAR T-cell-mediated immune response in a patient in need thereof, the method comprising: (i) contacting CAR T-cells with an effective amount of an interleukin-2-inducible T-cell kinase inhibitor in vitro, thereby producing ITK-contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK-contacted CAR T- cells; thereby stimulating the CAR T-cell-mediated immune response in the patient.

16. A method for increasing T-cell function in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2-inducible T-cell kinase inhibitor; thereby increasing T-cell function in the patient.

17. A method for increasing CAR T-cell function in a patient in need thereof, themethod comprising administering to the patient an effective amount of an interleukin-2- inducible T-cell kinase inhibitor; wherein the patient is receiving or has received CAR T-cell therapy; thereby increasing CAR T-cell function in the patient.

18. A method for increasing CAR T-cell function in a patient in need thereof, the method comprising administering to the patient: (i) an effective amount of an interleukin-2- inducible T-cell kinase inhibitor and (ii) an effective amount of CAR T-cells, thereby increasing CAR T-cell function in the patient.

19. A method for increasing CAR T-cell function in a patient in need thereof, the method comprising: (i) contacting CAR T-cells with an effective amount of an interleukin-2- inducible T-cell kinase inhibitor in vitro, thereby producing ITK-contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK-contacted CAR T-cells; thereby increasing CAR T-cell function in the patient.

20. A method of increasing expression of T cell factor-1 on T-cells in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2-inducible T-cell kinase inhibitor; thereby increasing expression of T cell factor-1 on T-cells in the patient.

21. A method for increasing expression of T cell factor-1 on CAR T-cells in a patient in need thereof, the method comprising administering to the patient an effective amount of an interleukin-2-inducible T-cell kinase inhibitor; wherein the patient is receiving or has received CAR T-cell therapy; thereby increasing expression of T cell factor-1 on CAR T-cells in the patient.

22. A method of increasing expression of T cell factor-1 on CAR T-cells in a patient in need thereof, the method comprising administering to the patient (i) an effective amount of an interleukin-2-inducible T-cell kinase inhibitor and (ii) an effective amount of CAR T-cells, thereby increasing expression of T cell factor-1 on CAR T-cells in the patient.

23. A method for increasing expression of T-cell factor-1 on CAR T-cells in a patient in need thereof, the method comprising: (i) contacting CAR T-cells with an effective amount of an interleukin-2-inducible T-cell kinase inhibitor in vitro, thereby producing ITK-contacted CAR T-cells; and (ii) administering to the patient an effective amount of the ITK-contacted CAR T-cells; thereby increasing expression of T-cell factor-1 on CAR T-cells in the patient.

24. The method of claim 1, further comprising measuring an increased level of TIM3, an increased level of LAG3, an increased level of TIGIT, an increased level of PD-1, adecreased level of IFNγ, a decreased level of granzyme B, or a combination of two or more thereof, relative to a control, in a biological sample obtained from the patient.

25. A method for making CAR T-cells, the method comprising contacting an interleukin-2-inducible T-cell kinase inhibitor with CAR T-cells in vitro.

26. A method for increasing expression of T-cell factor-1 on CAR T-cells, the method comprising contacting an interleukin-2-inducible T-cell kinase inhibitor with CAR T- cells in vitro, thereby increasing expression of T-cell factor-1 on CAR T-cells.

27. A method for increasing CAR T-cell function, the method comprising contacting an interleukin-2-inducible T-cell kinase inhibitor with CAR T-cells in vitro, thereby increasing CAR T-cell function.

28. A method of reducing CAR T-cell exhaustion, the method comprising contacting an interleukin-2-inducible T-cell kinase inhibitor with CAR T-cells in vitro, thereby reducing CAR T-cell exhaustion.

29. The method of claim 1, wherein the CAR T-cells express a chimeric antigen receptor comprising an extracellular domain that binds to an antigen on a cancer cell, wherein the antigen is epidermal growth factor receptor, mesothelin, prostate-specific membrane antigen, disialoganglioside GD2, interleukin-13Ra2, glypican-3, carbonic anhydrase IX, L1CAM, fibroblast activation protein, a cancer testis antigen, MUC1, FOLR1, BCMA, CD133, alpha-fetoprotein, carcinoembryonic antigen, trophoblast glycoprotein precursor, immature lamininreceptor protein, fetal sulphoglycoprotein, g-fetoprotein, cc2-H-ferroportin, beta-2- microglobulin, beta-human chorionic gonadotropin, calcitonin, bladder tumor antigen, CD7, CD19, CD117, CA15-3, CA19-9, CA-125, CA 27.29, CA72-4, CD20, CD22, CD25, CD30, CD33, chromogranin A, cytokeratin fragment 21-1, estrogen receptor, progesterone receptor, fibrinogen, gastrin, HE4, Her2 / neu, neuron-specific enolase, nuclear matrix protein 22, prostatic acid phosphatase, PD-L1, PD-1, prostate-specific antigen, somatostatin receptor, thyroglobulin, 5-HIAA, osteocalcin, transferrin recepto, alkaline phosphtase, BRAF, KRAS, NMP22, BRCA2, urokinase plasminogen activator, apoliprotein A1, S100, nestin, cytokeratin fragments 21-1, ferritin, tissue polypeptide antigen, epididymal secretory protein E4, CECAM 5, CECAM 6, serum M-protein, CTLA-4, B7-1 / B7-2, epithelial tumor antigen, tyrosinase, melanoma- associated antigen, p53, mutated p53, MART-2, beta-catenin, mutated ras, mutated KRAS, BAGE, GAGE-1, NY-ESO-1, PRAME, CT83, SSX2, ART-4, BAGE, beta-catenin / m, Bcr-abL CAMEL, CAP-1 , CASP-8, CDC27 / m, CD 4 / m, a cell surface protein of the claudin family, c-MYC, CT, Cyp-B, DAM, ELF2M, ETV6-AML1, G250, GAGE, GnT-V, KPL-1, HAGE, HPV- E7, HPV-E6, HAST-2, hTERT, LAGE, LDLR / FUT, MAGE- A, MAGE-B, MAGE-C, MART- 1, Melan-A, MCIR-1, myosin, MUM-1, MUM-2, MUM-3, NA88-A, NF1, NY-ESO-1, NY-BR- 1, HSP90, BCR-ABL-1, RAR, PRAME, proteinase 3, PSM, RAGE, RU1, RU2, SAGE, SART- 1, SART-3, SCGB3A2, SCP 1, SCP2, SCP3, SSX, Survrvin, TEL / AML1, TPEM, TRP-1, TRP- 2, TRP-2-INT2, TPTE, WT, or a combination of two or more thereof.

30. The method of claim 1, wherein the CAR T-cells are a CAR T-cell therapyselected from the group consisting of axicabtagene ciloleucel, brexucabtagene autoleucel,ciltacabtagene autoleucel, idecabtagene vicleucel, lisocabtagene maraleucel, andtisagenlecleucel.

31. The method of claim 1, wherein the interleukin-2-inducible T-cell kinase inhibitor is soquelitinib or a pharmaceutically acceptable salt thereof.

32. A kit comprising an interleukin-2-inducible T-cell kinase inhibitor and CAR T- cells.

33. A composition comprising an interleukin-2-inducible T-cell kinase inhibitor and CAR T-cells.

34. The composition of claim 33, wherein the interleukin-2-inducible T-cell kinase inhibitor is soquelitinib or a pharmaceutically acceptable salt thereof.

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