WO2025097025A1 - Protein kinase c (pkc) theta inhibitor compounds - Google Patents

Abstract

The present disclosure provides protein kinase C (PKC) theta inhibitor compounds. Also provided by the present disclosure are methods of use of such compounds, as well as pharmaceutical compositions that include the PKC theta inhibitor compounds. The PKC theta inhibitor compounds and pharmaceutical compositions thereof find use in the treatment of PKC theta related conditions.

Description

PROTEIN KINASE C (PKC) THETA INHIBITOR COMPOUNDS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 63/547,300, filed November 3, 2023, the disclosure of which is incorporated by reference herein in its entirety.

INTRODUCTION

[0002] Protein kinases are a large family of structurally related enzymes that are responsible for the control of a variety of signal transduction processes within the cell (see Hardie, G and Hanks, S. The Protein Kinase Facts Book, I and II, Academic Press, San Diego, Calif.: 1995).

[0003] Protein kinase C-theta (PKC-theta), a serine/threonine kinase, is a member of the novel, calcium independent PKC subfamily that is selectively expressed in T cells and skeletal muscle. PKC-theta has been shown to interact with several different biochemical processes, including T cell activation (Monks et al., 1997, Nature, 385: 83-86; Monks et al., 1998, Nature, 395: 82-86) and autoimmune diseases (Salek-Ardakani et al., 2006; Tan et al., 2006; Healy et al., 2006; Anderson et al., 2006; Nagahama et al., 2008). As such, PKC-theta inhibitor compounds could be useful for treating various medical diseases and conditions.

SUMMARY

[0004] Provided are protein kinase C (PKC) theta inhibitor compounds. Also provided are methods of use of such compounds, as well as pharmaceutical compositions thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

[0005] FIG. 1 shows a plate map corresponding to a cell assay procedure as per the present disclosure.

[0006] FIG. 2 shows a plate map corresponding to an enzyme assay procedure as per the present disclosure.

[0007] FIG. 3 shows a graph of the results from an sRICA assay as per the present disclosure.

[0008] FIG. 4 shows a graph of the results from an sRICA assay as per the present disclosure,

[0009] FIG. 5 shows in vivo mouse data of compound as per the present disclosure,

[0010] FIG. 6 shows in vivo mouse data of compound as per the present disclosure.

[0011] FIG. 7 shows in vivo mouse data of compound as per the present disclosure.

[0012] FIG. 8 shows in vivo mouse data of compound as per the present disclosure.

[0013] FIG. 9 shows in vivo mouse data of compound as per the present disclosure.

[0014] FIG. 10 shows in vivo mouse data of compound as per the present disclosure, DETAILED DESCRIPTION

[0015] Provided are protein kinase C (PKC) theta inhibitor compounds. Also provided are methods of use of such compounds, as well as pharmaceutical compositions thereof.

[0016] Before the present invention is described in greater detail, it is to be understood that this invention is not limited to particular embodiments described, as such may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0017] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limits of that range is also specifically disclosed. Each smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range, and each range where either, neither or both limits are included in the smaller ranges is also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

[0018] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, some potential and exemplary methods and materials may now be described. Any and all publications mentioned herein are incorporated herein by reference to disclose and describe the methods and/or materials in connection with which the publications are cited. It is understood that the present disclosure supersedes any disclosure of an incorporated publication to the extent there is a contradiction.

[0019] It must be noted that as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a droplet" includes a plurality of such droplets and reference to "the discrete entity" includes reference to one or more discrete entities, and so forth. It is further noted that the claims may be drafted to exclude any element, e.g., any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as “solely”, “only” and the like in connection with the recitation of claim elements, or the use of a “negative” limitation. [0020] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed. To the extent the definition or usage of any term herein conflicts with a definition or usage of a term in an application or reference incorporated by reference herein, the instant application shall control.

[0021] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present invention. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.

DEFINITIONS

[0022] "Alkyl" refers to a monoradical, branched or linear, non-cyclic, saturated hydrocarbon group. Exemplary alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t- butyl, octyl, decyl, cyclopentyl, and cyclohexyl. In some cases, the alkyl group has 1 to 24 carbon atoms, e.g. 1 to 12, 1 to 6, or 1 to 3.

[0023] “Alkenyl" refers to a monoradical, branched or linear, non-cyclic hydrocarbonyl group that comprises a carbon-carbon double bond. Exemplary alkenyl groups include ethenyl, n- propenyl, isopropenyl, n-butenyl, isobutenyl, octenyl, decenyl, tetradecenyl, hexadecenyl, eicosenyl, and tetracosenyl.

[0024] “Alkynyl" refers to a monoradical, branched or linear, non-cyclic hydrocarbonyl group that comprises a carbon-carbon triple bond. Exemplary alkynyl groups include ethynyl and n- propynyl.

[0025] “Cycloalkyl” refers to a monoradical, cyclic, saturated hydrocarbon group. Exemplary cycloalkyl groups include cyclooctyl and bicyclopentyl. Similarly, “cycloalkenyl” refers to a monoradical and cyclic group having carbon-carbon double bond whereas “cycloalkynyl” refers to a monoradical and cyclic group having carbon-carbon triple bond.

[0026] “Heterocyclyl” refers to a monoradical, cyclic group that contains a heteroatom (e.g. O, S, N) as a ring atom and that is not aromatic (i.e. distinguishing heterocyclyl groups from heteroaryl groups). Exemplary heterocyclyl groups include piperidinyl, tetrahydrofuranyl, dihydrofuranyl, and thiocanyl.

[0027] “Aryl" refers to an aromatic group containing at least one aromatic ring, wherein each of the atoms in the ring are carbon atoms, i.e. none of the ring atoms are heteroatoms (e.g. O, S, N). In some cases the aryl group has a second aromatic ring, e.g. that is fused to the first aromatic ring. Exemplary aryl groups are phenyl, naphthyl, biphenyl, diphenylether, diphenylamine, and benzophenone.

[0028] “Heteroaryl” refers to an aromatic group containing at least one aromatic ring, wherein at least one of the atoms in the aromatic ring is a heteroatom (e.g. O, S, N). Exemplary heteroaryl groups include those obtained from removing a hydrogen atom from pyridine, pyrimidine, furan, thiophene, or benzothiophene.

[0029] The term “substituted” refers the removal of one or more hydrogens from an atom (e.g. from a C or N atom) and their replacement with a different group. For instance, a hydrogen atom on a phenyl (-CeHs) group can be replaced with a methyl group to form a -C6H4CH3 group. Thus, the -C6H4CH3 group can be considered a substituted aryl group. As another example, two hydrogen atoms from the second carbon of a propyl (-CH2CH2CH3) group can be replaced with an oxygen atom to form a -CH2C(O)CH3 group, which can be considered a substituted alkyl group. However, replacement of a hydrogen atom on a propyl (-CH2CH2CH3) group with a methyl group (e.g. giving -CH2CH(CH3)CH3) is not considered a “substitution” as used herein since the starting group and the ending group are both alkyl groups. However, if the propyl group was substituted with a methoxy group, thereby giving a -CH2CH(OCH3)CH3 group, the overall group can no long be considered “alkyl”, and thus is “substituted alkyl”. Thus, in order to be considered a substituent, the replacement group is a different type than the original group. In addition, groups are presumed to be unsubstituted unless described as substituted. For instance, the term “alkyl” and “unsubstituted alkyl” are used interchangeably herein.

[0030] Exemplary substituents include alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof.

[0031] In some cases, the substitutions can themselves be further substituted with one or more groups. For example, the group -C6H4CH2CH3 can be considered as substituted aryl, i.e. an aryl group substituted with the ethyl, which is an alkyl group. Furthermore, the ethyl group can itself be substituted with a pyridyl group to form -C6H4CH2CH2C5H5N, wherein -C6H4CH2CH2C5H5N can also be considered as a substituted aryl group as the term is used herein. In some cases, the substituents are not substituted with any other groups.

[0032] Diradical groups are also described herein, i.e. in contrast to the monoradical groups such as alkyl and aryl described above. The term "alkylene" refers to the diradical version of an alkyl group, i.e. an alkylene group is a diradical, branched or linear, cyclic or non-cyclic, saturated hydrocarbon group. Exemplary alkylene groups include diylmethane (-CH2-, which is also known as a methylene group), 1 ,2-diylethane (-CH2CH2-), and 1,1-diylethane (i.e. a CHCH3 fragment where the first atom has two single bonds to other two different groups). The term “arylene” refers to the diradical version of an aryl group, e.g. 1,4-diylbenzene refers to a C6H4 fragment wherein two hydrogens that are located para to one another are removed and replaced with single bonds to other groups. The terms “alkenylene”, “alkynylene”, “heteroarylene”, and “heterocyclene” are also used herein.

[0033] “Acyl” refers to a group of formula -C(O)R wherein R is alkyl, alkenyl, alkynyl, or substituted versions thereof. For example, the acetyl group has formula -C(O)CH3. “Carbonyl” refers to a diradical group of formula -C(O)-.

[0034] “Alkoxy" refers to a group of formula -O(alkyl). Similar groups can be derived from alkenyl, alkynyl, aryl, heteroaryl, and other groups.

[0035] “Amino" refers to the group -NR^XR^Y wherein R^x and R^Y are each independently H or a non-hydrogen substituent. Exemplary non-hydrogen substituents include alkyl groups (e.g. methyl, ethyl, and isopropyl).

[0036] “Carbonyl” refers to a diradical group of formula -C(O)-.

[0037] “Carboxy” is used interchangeably with carboxyl and carboxylate to refer to the -CO2H group and salts thereof.

[0038] “Cyano” or “nitrile” refers to the group -CN.

[0039] “Ether” refers to a diradical group of formula -O-. For instance, if the ether group is connected to an alkyl group, then the overall group is an alkoxy group (e.g. -OCH3 or methoxy). If the ether is connected to a carbonyl group, then the overall group is an ester group of formula - OC(O)-.

[0040] “Halo” and “halogen” refer to the chloro, bromo, fluoro, and iodo groups.

[0041] “Haloalkyl” refers to an alkyl group that is substituted with at least one halogen. Stated in another manner, a haloalkyl group is a substituted alkyl group wherein at least one substituent is a halogen. Trifluoromethyl is an exemplary haloalkyl group.

[0042] “Nitro” refers to the group of formula -NO2.

[0043] “Bicyclic” refers to compounds containing more than one ring with at least two common atoms. For example, the rings may be fused, as in naphthalene, or separate, as in biphenyl.

[0044] Unless otherwise specified, reference to an atom is meant to include all isotopes of that atom. For example, reference to H includes ¹ H, ²H (i.e. D or deuterium) and ³H (i.e. tritium), and reference to C is includes both ¹²C and all other isotopes of carbon (e.g. ¹³C). Unless specified otherwise, groups include all possible stereoisomers.

COMPOUNDS OF FORMULA (I)

[0045] Provided are PKC-theta inhibitor compounds of formula (I):

wherein:

Z is CR³ or N;

X is a bicyclic and optionally substituted heteroaryl group;

R¹ and R² are each independently selected from the group consisting of H, alkyl, halo, amine, cyano and hydroxyl, or R¹ and R² together with the atoms to which they are attached form a 5-membered or a 6-membered aryl or a heteroaryl ring;

R³ is selected from the group consisting of H and halo; and

R⁴ and R⁵ are each independently selected from the group consisting of H, C2-4 linear alkyl, C2-4 branched alkyl and C3-6 cycloalkyl group.

[0046] In certain embodiments, R¹ and R² are each independently selected from the group consisting of H, alkyl, halo, amine, cyano and hydroxyl.

[0047] In some cases, R¹ is H. In some cases, R¹ is alkyl. For instance, in some cases, R¹ is methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec -butyl, tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl or decyl. In some cases, R¹ is halo. For instance, in some cases, R¹ is fluoro (F), chloro (Cl), bromo (Br) or iodo (I).

[0048] In some cases, R¹ is amine. In some cases, R¹ is cyano. In some cases, R¹ is hydroxyl.

[0049] In some cases, R² is H. In some cases, R² is alkyl. For instance, in some cases, R² is methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec -butyl, tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl or decyl. In some cases, R² is halo. For instance, in some cases, R² is fluoro (F), chloro (Cl), bromo (Br) or iodo (I).

[0050] In some cases, R² is amine. In some cases, R²is cyano. In some cases, R² is hydroxyl.

[0051] In certain embodiments, R¹ is fluoro and R² is H. In some cases, R¹ is chloro and R² is

H. In some cases, R¹ is amine and R² is H.

[0052] As shown in formula (I), R¹ and R² together with the atoms to which they are attached form a 5-membered ring or a 6-membered aryl ring or a heteroaryl ring. In some cases, R¹ and R² together with the atoms to which they are attached form a 5-membered ring. In some cases, the 5- membered ring is selected from the group consisting of pyrrole, pyrroline, pyrazole, imidazole, triazole and substituted versions thereof. [0053] In some cases, R¹ and R² together with the atoms to which they are attached form a 6- membered aryl ring. In some cases, R¹ and R² together with the atoms to which they are attached form a 6-membered heteroaryl ring. For instance, in some cases, the 6-membered ring is selected from the group consisting of pyridine, pyridazine, pyrimidine, pyrazine, triazine and substituted versions thereof.

[0054] In some cases, R¹ is -NH- and R² is -CH-. In some cases, R¹ is -CH- and R² is -NH-.

[0055] In certain embodiments, Z is CR³ or N. In some cases, Z is CR³. In some cases, Z is N.

When Z is CR³, R³ is selected from the group consisting of H and halo.

[0056] In certain embodiments, R³ is halo. For instance, in some cases, R³ is fluoro (F), chloro (Cl), bromo (Br) or iodo (I). In certain embodiments, R³ is fluoro.

[0057] In some cases, R⁴ and R⁵ are each independently selected from the group consisting of H, C2-4 linear alkyl, C2-4 branched alkyl and C3-6 cycloalkyl group.

[0058] In some cases, R⁴ is H. In some cases, R⁴ is C2-4 linear alkyl. For instance, in some cases, R⁴ is ethyl, propyl, or butyl.

[0059] In some cases, R⁴ is C2-4 branched alkyl. For instance, in some cases, R⁴ is isopropyl, isobutyl, sec -butyl, or tert-butyl. In some cases, R⁴ is isopropyl.

[0060] In some cases, R⁵ is H. In some cases, R⁵is C2-4 linear alkyl. For instance, in some cases, R⁵ is ethyl, propyl, or butyl.

[0061] In some cases, R^s is C2-4 branched alkyl. For instance, in some cases, R^s is isopropyl, isobutyl, sec -butyl, or tert-butyl. In some cases, R⁵ is isopropyl.

[0062] In some cases, R⁴ is cyclopropane, cyclobutene, cyclopentane and cyclohexane. In some cases, R³ is cyclopropane, cyclobutene, cyclopentane and cyclohexane.

[0063] As described above, X is a bicyclic and optionally substituted heteroaryl group. Since X is bicyclic, X includes a first ring and a second ring. In some cases, X has the structure:

wherein: ring A is a 5-membered and optionally substituted heteroaryl group; and ring B is a 6-membered and optionally substituted heteroaryl group.

[0064] As such, ring A is the part of group X that is covalently bonded to the pyridine ring of formula (I), wherein ring A is fused to ring B. As used herein, the term “optionally substituted heteroaryl group” means that in some cases the group is a substituted heteroaryl group and in other cases the group is an unsubstituted heteroaryl group. In some cases, the heteroaryl group is substituted with one or more groups selected from the group consisting of halo, alkyl, and oxo.

[0065] In some embodiments, ring A is selected from the group consisting of

[0066] In some embodiments, ring B is selected from the group consisting of pyridine, pyridazine, pyrimidine, pyrazine, cycloalkyl, and substituted versions thereof. The recitation of the option of substituted versions thereof means that the group can be pyridine, substituted pyridine, pyridazine, substituted pyridazine, pyrimidine, substituted pyrimidine, pyrazine, substituted pyrazine, cycloalkyl, and substituted cycloalkyl. In some cases, the group is a substituted cycloalkyl group of bicyclopentyl.

[0067] In some embodiments, the compounds are of formula (la):

wherein:

Z is CR³ or N;

X is a bicyclic and optionally substituted heteroaryl group;

R¹ is -CH-;

R² is -NH-;

R³ is selected from the group consisting of H and halo; and

R⁴ and R⁵ are each independently selected from the group consisting of H, C2-4 linear alkyl, C2-4 branched alkyl and C3-6 cycloalkyl group.

[0068] In certain embodiments, Z is CR³ or N. In some cases, Z is CR³. In some cases, Z is N. When Z is CR³, R³ is selected from the group consisting of H and halo.

[0069] In certain embodiments, R³ is H. In certain embodiments, R³ is halo. For instance, in some cases, R³ is fluoro (F), chloro (Cl), bromo (Br) or iodo (I). In certain embodiments, R³ is fluoro.

[0070] In some cases, R¹ is -CH- and R² is -NH-.

[0071] In some embodiments of formula (la), R⁴, R^s and X are as defined above.

[0072] In some embodiments, the compound is of formula (lb):

[0073] In certain embodiments, Z is CR³ or N. In some cases, Z is CR³. In some cases, Z is N. When Z is CR³, R³ is selected from the group consisting of H and halo.

[0074] In certain embodiments, R³ is H. In certain embodiments, R³ is halo. For instance, in some cases, R³ is fluoro (F), chloro (Cl), bromo (Br) or iodo (I). In certain embodiments, R³ is fluoro.

[0075] In some cases, R¹ is -NH-, -S-, -O-, or -N-lower alkyl. In some cases, R¹ is -NH-. In some cases, R¹ is -S-. In some cases, R¹ is -O-. In some cases, R¹ is -N-lower alkyl.

[0076] In some cases, R² is -CH-, -NH-, -S-, -O-, or -N-lower alkyl. In some cases, R² is -CH-. In some cases, R² is -NH-. In some cases, R² is -S-. In some cases, R² is -O-. In some cases, R² is -N-lower alkyl.

[0077] In some embodiments of formula (lb), R⁴, R^s and X are as defined above.

[0078] In some embodiments, the compound is of formula (1c):

[0079] In certain embodiments, R¹ and R² are each independently selected from the group consisting of H, alkyl, halo, amine, cyano and hydroxyl.

[0080] In some cases, R¹ is H. In some cases, R¹ is alkyl. For instance, in some cases, R¹ is methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec -butyl, tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl or decyl. In some cases, R¹ is halo. For instance, in some cases, R¹ is fluoro (F), chloro (Cl), bromo (Br) or iodo (I).

[0081] In some cases, R¹ is amine. In some cases, R¹ is cyano. In some cases, R¹ is hydroxyl.

[0082] In some cases, R² is H. In some cases, R² is alkyl. For instance, in some cases, R² is methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec -butyl, tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl or decyl. In some cases, R² is halo. For instance, in some cases, R² is fluoro (F), chloro (Cl), bromo (Br) or iodo (I). [0083] In some cases, R² is amine. In some cases, R² is cyano. In some cases, R² is hydroxyl.

[0084] In certain embodiments, R¹ is fluoro and R² is H. In some cases, R¹ is chloro and R² is H. In some cases, R¹ is amine and R² is H.

[0085] As shown in formula (II), R¹ and R² together with the atoms to which they are attached form a 5-membered ring or a 6-membered aryl ring or a heteroaryl ring. In some cases, R¹ and R² together with the atoms to which they are attached form a 5-membered ring. In some cases, the 5- membered ring is selected from the group consisting of pyrrole, pyrroline, pyrazole, imidazole, triazole and substituted versions thereof.

[0086] In some cases, R¹ and R² together with the atoms to which they are attached form a 6- membered aryl ring. In some cases, R¹ and R² together with the atoms to which they are attached form a 6-membered heteroaryl ring. For instance, in some cases, the 6-membered ring is selected from the group consisting of pyridine, pyridazine, pyrimidine, pyrazine, triazine and substituted versions thereof.

[0087] In some cases, R¹ is -NH- and R² is -CH-. In some cases, R¹ is -CH- and R² is -NH-.

[0088] In certain embodiments, Z is CR³ or N. In some cases, Z is CR³. In some cases, Z is N. When Z is CR³, R³ is selected from the group consisting of H and halo.

[0089] In certain embodiments, R³ is halo. For instance, in some cases, R³ is fluoro (F), chloro (Cl), bromo (Br) or iodo (I). In certain embodiments, R³ is fluoro.

[0090] In some cases, R⁴ and R^s are each independently selected from the group consisting of H, C2-4 linear alkyl, C2-1 branched alkyl and C3-6 cycloalkyl group.

[0091] In some cases, R⁴ is H. In some cases, R⁴ is C2-4 linear alkyl. For instance, in some cases, R⁴ is ethyl, propyl, or butyl.

[0092] In some cases, R⁴ is C2-4 branched alkyl. For instance, in some cases, R⁴ is isopropyl, isobutyl, sec -butyl, or tert-butyl. In some cases, R⁴ is isopropyl.

[0093] In some cases, R⁵ is H. In some cases, R⁵is C2-4 linear alkyl. For instance, in some cases, R⁵ is ethyl, propyl, or butyl.

[0094] In some cases, R^s is C2-4 branched alkyl. For instance, in some cases, R^s is isopropyl, isobutyl, sec-butyl, or tert-butyl. In some cases, R⁵ is isopropyl.

[0095] In some cases, R⁴ is cyclopropane, cyclobutene, cyclopentane and cyclohexane. In some cases, R⁵ is cyclopropane, cyclobutene, cyclopentane and cyclohexane.

[0096] As described above, X is a bicyclic and optionally substituted heteroaryl group. Since X is bicyclic, X includes a first ring and a second ring. In some cases, X has the structure:

wherein: ring A is a 5-membered and optionally substituted heteroaryl group; and ring B is a 6-membered and optionally substituted heteroaryl group.

[0097] As such, ring A is the part of group X that is covalently bonded to the pyridine ring of formula (I), wherein ring A is fused to ring B. As used herein, the term “optionally substituted heteroaryl group” means that in some cases the group is a substituted heteroaryl group and in other cases the group is an unsubstituted heteroaryl group. In some cases, the heteroaryl group is substituted with one or more groups selected from the group consisting of halo, alkyl, and oxo.

[0098] In some embodiments, ring A is selected from the group consisting of

[0099] In some embodiments, ring B is selected from the group consisting of pyridine, pyridazine, pyrimidine, pyrazine, cycloalkyl, and substituted versions thereof. The recitation of the option of substituted versions thereof means that the group can be pyridine, substituted pyridine, pyridazine, substituted pyridazine, pyrimidine, substituted pyrimidine, pyrazine, substituted pyrazine, cycloalkyl, and substituted cycloalkyl. In some cases the group is a substituted cycloalkyl group of bicyclopentyl.

[00100] In some embodiments, the compound is of formula (Id):

[00101] In certain embodiments, Z is CR³ or N. In some cases, Z is CR³. In some cases, Z is N. When Z is CR³, R is selected from the group consisting of H and halo.

[00102] In certain embodiments, R³ is halo. For instance, in some cases, R³ is fluoro (F), chloro (Cl), bromo (Br) or iodo (I). In certain embodiments, R³ is fluoro.

[00103] In some cases, R¹ is -CH- and R² is -NH-.

[00104] In some embodiments of formula (la), R⁴, R⁵ and X are as defined above.

[00105] In some embodiments, the compound is of formula (le):

(le).

[00106] In certain embodiments, Z is CR³ or N. In some cases, Z is CR³. In some cases, Z is N. When Z is CR³, R³ is selected from the group consisting of H and halo.

[00107] In certain embodiments, R³ is halo. For instance, in some cases, R³ is fluoro (F), chloro (Cl), bromo (Br) or iodo (I). In certain embodiments, R³ is fluoro.

[00108] In some cases, R¹ is -NH- and R² is -CH-.

[00109] In some embodiments of formula (lb), R⁴, R⁵ and X are as defined above.

[00110] In some embodiments, the compounds can be selected from list in Table 1 below.

[00111] Table 1:

COMPOSITIONS ND FORMULATIONS

[00112] Also provided are compositions comprising a compound according to formula (I), formula (la), formula (lb), formula (Ic), formula (Id) or formula (le), including all sub-generic formulae if any, as described above. In some cases, the composition includes a racemic mixture of stereoisomers. In some embodiments, the composition is enriched in a particular stereoisomer, e.g., the composition is enriched in a first enantiomer relative to a second enantiomer. The term “enantiomeric excess” is used herein to quantify the relative amount of the first enantiomer compared to the second enantiomer, wherein enantiomeric excess is the absolute difference between the mole fraction of each enantiomer. For instance, if 70% of a compound is a first enantiomer and 30% of the compound is the second enantiomer, then the enantiomeric excess is 40% (i.e., 70% minus 40%). In some cases, the composition has an enantiomeric excess of the first enantiomer of 1% or more, such as 10% or more, 20% or more, 30% or more, or 40% or more. In some embodiments, the composition is an aqueous solution of the PKC theta inhibitor compound.

[00113] In certain embodiments, the disclosed compounds and prodrugs thereof are useful for the treatment of a disease or disorder. Accordingly, pharmaceutical compositions comprising at least one disclosed compound or prodrug are also described herein. For example, the present disclosure provides pharmaceutical compositions that include a therapeutically effective amount of a compound or prodrug of the present disclosure (or a pharmaceutically acceptable salt or solvate or hydrate or stereoisomer thereof) and a pharmaceutically acceptable excipient.

[00114] A pharmaceutical composition that includes a subject compound (or prodrug) may be administered to a patient alone, or in combination with other supplementary active agents. For example, one or more compounds or prodrugs according to the present disclosure can be administered to a patient with or without supplementary active agents. The pharmaceutical compositions may be manufactured using any of a variety of processes, including, but not limited to, conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, lyophilizing, and the like. The pharmaceutical composition can take any of a variety of forms including, but not limited to, a sterile solution, suspension, emulsion, spray dried dispersion, lyophilisate, tablet, microtablets, pill, pellet, capsule, powder, syrup, elixir or any other dosage form suitable for administration.

[00115] A compound or prodrug of the present disclosure may be administered to a subject using any convenient means capable of resulting in the desired reduction in disease condition or symptom. Thus, a compound or prodrug can be incorporated into a variety of formulations for therapeutic administration. More particularly, a compound or prodrug can be formulated into pharmaceutical compositions by combination with appropriate pharmaceutically acceptable excipients, carriers or diluents, and may be formulated into preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, creams, gels, foams, solutions, suppositories, injections, inhalants, aerosols, and the like.

[00116] Formulations for pharmaceutical compositions are described in, for example, Remington’s Pharmaceutical Sciences, by E. W. Martin, Mack Publishing Co., Easton, Pa., 19th Edition, 1995, which describes examples of formulations (and components thereof) suitable for pharmaceutical delivery of the disclosed compounds. Pharmaceutical compositions that include at least one of the compounds or prodrugs can be formulated for use in human or veterinary medicine. Particular formulations of a disclosed pharmaceutical composition may depend, for example, on the mode of administration and/or on the location of the subject to be treated. In some embodiments, formulations include a pharmaceutically acceptable excipient in addition to at least one active ingredient, such as a compound of the present disclosure. In other embodiments, other medicinal or pharmaceutical agents, for example, with similar, related or complementary effects on the disease or condition being treated can also be included as active ingredients in a pharmaceutical composition.

[00117] Pharmaceutically acceptable carriers useful for the disclosed methods and compositions may depend on the particular mode of administration being employed. In addition to biologically neutral carriers, pharmaceutical compositions to be administered can optionally contain non-toxic auxiliary substances (e.g., excipients), such as wetting or emulsifying agents, preservatives, and pH buffering agents, and the like. The disclosed pharmaceutical compositions may be formulated as a pharmaceutically acceptable salt of a disclosed compound.

[00118] The term “unit dosage form,” as used herein, refers to physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of a compound or prodrug calculated in an amount sufficient to produce the desired effect in association with a pharmaceutically acceptable diluent, excipient, carrier or vehicle. The specifications for a compound or prodrug depend on the particular compound or prodrug employed and the effect to be achieved, and the pharmacodynamics associated with each compound in the subject.

[00119] The dosage form of a disclosed pharmaceutical composition may be determined by the mode of administration chosen. For example, in addition to injectable fluids, topical or oral dosage forms may be employed. Topical preparations may include eye drops, ointments, sprays and the like. Oral formulations may be liquid (e.g., syrups, solutions or suspensions), or solid (e.g., powders, pills, tablets, or capsules). Methods of preparing such dosage forms are known, or will be apparent, to those skilled in the art.

[00120] Certain embodiments of the pharmaceutical compositions that include a subject compound or prodrug may be formulated in unit dosage form suitable for individual administration of precise dosages. The amount of active ingredient administered may depend on the subject being treated, the severity of the affliction, and the manner of administration, and is known to those skilled in the art. In certain instances, the formulation to be administered contains a quantity of the compound or prodrug disclosed herein in an amount effective to achieve the desired effect in the subject being treated.

[00121] Each therapeutic compound can independently be in any dosage form, such as those described herein, and can also be administered in various ways, as described herein. For example, the compounds or prodrugs may be formulated together, in a single dosage unit (that is, combined together in one form such as capsule, tablet, powder, or liquid, etc.) as a combination product. Alternatively, when not formulated together in a single dosage unit, an individual compound or prodrug may be administered at the same time as another therapeutic compound or sequentially, in any order thereof.

[00122] A disclosed compound can be administered alone, as the sole active pharmaceutical agent, or in combination with one or more additional compounds or prodrugs of the present disclosure or in conjunction with other agents. When administered as a combination, the therapeutic agents can be formulated as separate compositions that are administered simultaneously or at different times, or the therapeutic agents can be administered together as a single composition combining two or more therapeutic agents. Thus, the pharmaceutical compositions disclosed herein containing a compound of the present disclosure optionally include other therapeutic agents. Accordingly, certain embodiments are directed to such pharmaceutical compositions, where the composition further includes a therapeutically effective amount of an agent selected as is known to those of skill in the art.

METHODS OF TREATMENT

[00123] Provided are methods of treating a subject for a condition by administering a PKC theta inhibitor compound as described herein. In some cases, the subject has been diagnosed with a PKC-theta related condition. For example, the condition could be related to T cell activation, e.g. an autoimmune disease.

[00124] Nonlimiting examples of the PKC-theta related condition are provided below and elsewhere. It is expected that the ordinarily skilled artisan will be able to extrapolate from these examples to other indications and biotherapeutics as known in the art. For example, the individual may be suffering from an autoimmune disease, e.g. inflammatory bowel disease, Crohn’s disease, ulcerative colitis, psoriasis, atopic dermatitis, rheumatoid arthritis, multiple sclerosis, type-1 diabetes, or graph versus host disease. In such instances, the method may include administering to the individual a PKC theta inhibitor compound as disclosed herein in an amount effective to treat the autoimmune disease. Accordingly, aspects of the present disclosure include methods of treating an autoimmune disease, such as inflammatory bowel disease, Crohn’s disease, ulcerative colitis, psoriasis, atopic dermatitis, rheumatoid arthritis, multiple sclerosis, type-1 diabetes, or graph versus host disease, by administering to an individual in need thereof a therapeutically effective amount of a PKC theta inhibitor compound as disclosed herein.

[00125] By “treatment” is meant that at least an amelioration of the symptoms associated with the condition afflicting the host is achieved, where amelioration is used in a broad sense to refer to at least a reduction in the magnitude of a parameter, e.g. symptom, associated with the condition being treated. As such, treatment also includes situations where the pathological condition, or at least symptoms associated therewith, are completely inhibited, e.g., prevented from happening, or stopped, e.g. terminated, such that the host no longer suffers from the condition, or at least the symptoms that characterize the condition. Thus treatment includes: (i) prevention, that is, reducing the risk of development of clinical symptoms, including causing the clinical symptoms not to develop, e.g., preventing disease progression to a harmful state; (ii) inhibition, that is, arresting the development or further development of clinical symptoms, e.g., mitigating or completely inhibiting an active disease; and/or (iii) relief, that is, causing the regression of clinical symptoms. [00126] The subject compounds or prodrugs find use for treating a disease or disorder in a subject. The route of administration may be selected according to a variety of factors including, but not limited to, the condition to be treated, the formulation and/or device used, the subject to be treated, and the like. Routes of administration useful in the disclosed methods include, but are not limited to, oral and parenteral routes, such as intravenous (iv), intraperitoneal (ip), rectal, topical, ophthalmic, nasal, otic, intrathecal, and transdermal. Formulations for these dosage forms are described herein.

[00127] An effective amount of a subject compound or prodrug may depend, at least, on the particular method of use, the subject being treated, the severity of the affliction, and the manner of administration of the therapeutic composition. A “therapeutically effective amount” of a composition is a quantity of a specified compound or prodrug sufficient to achieve a desired effect in a subject (e.g., patient) being treated. For example, this may be the amount of a subject compound necessary to prevent, inhibit, reduce or relieve a disease or disorder in a subject. Ideally, a therapeutically effective amount of a compound or prodrug is an amount sufficient to prevent, inhibit, reduce or relieve a disease or disorder in a subject without causing a substantial cytotoxic effect on host cells in the subject.

[00128] Therapeutically effective doses of a subject compound or prodrug or pharmaceutical composition can be determined by one of skill in the art. For example, in some instances, a therapeutically effective dose of a compound or prodrug or pharmaceutical composition is administered with a goal of achieving local (e.g., tissue) concentrations that are at least as high as the IC50 of an applicable compound disclosed herein. [00129] The specific dose level and frequency of dosage for any particular subject may be varied and may depend upon a variety of factors, including the activity of the subject compound or prodrug, the metabolic stability and length of action of that compound or prodrug, the age, body weight, general health, sex and diet of the subject, mode and time of administration, rate of excretion, drug combination, and severity of the condition of the host undergoing therapy.

[00130] In some embodiments, multiple doses of a compound or prodrug are administered. The frequency of administration of a compound can vary depending on any of a variety of factors, e.g., severity of the symptoms, condition of the subject, etc. For example, in some embodiments, a compound is administered once per month, twice per month, three times per month, every other week, once per week (qwk), twice per week, three times per week, four times per week, five times per week, six times per week, every other day, daily (qd/od), twice a day (bds/bid), or three times a day (tds/tid), etc.

EXAMPLES

[00131] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g. amounts, temperature, etc.) but ssome experimental errors and deviations should be accounted for. As will be understood, by those of skill in the art of organic synthesis and medicinal chemistry the specific conditions set forth below are exemplary and can be varied or adapted to other reagents and products in routine fashion. Unless indicated otherwise, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric. Standard abbreviations may be used, e.g., bp, base pair(s); kb, kilobase(s); pl, picoliter(s); s or sec, second(s); min, minute(s); h or hr, hour(s); aa, amino acid(s); nt, nucleotide(s); and the like.

General Synthetic Procedures

[00132] Many general references providing commonly known chemical synthetic schemes and conditions useful for synthesizing the disclosed compounds are available (see, e.g., Smith and March, March’s Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Fifth Edition, Wiley-Interscience, 2001; or Vogel, A Textbook of Practical Organic Chemistry, Including Qualitative Organic Analysis, Fourth Edition, New York: Longman, 1978).

[00133] Compounds as described herein can be purified by any purification protocol known in the art, including chromatography, such as HPLC, preparative thin layer chromatography, flash column chromatography and ion exchange chromatography. Any suitable stationary phase can be used, including normal and reversed phases as well as ionic resins. In certain embodiments, the disclosed compounds are purified via silica gel and/or alumina chromatography. See, e.g., Introduction to Modern Liquid Chromatography, 2nd Edition, ed. L. R. Snyder and J. J. Kirkland, John Wiley and Sons, 1979; and Thin Layer Chromatography, ed E. Stahl, Springer- Verlag, New York, 1969.

[00134] During any of the processes for preparation of the subject compounds, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned. This may be achieved by means of conventional protecting groups as described in standard works, such as J. F. W. McOmie, “Protective Groups in Organic Chemistry”, Plenum Press, London and New York 1973, in T. W. Greene and P. G. M. Wuts, “Protective Groups in Organic Synthesis”, Third edition, Wiley, New York 1999, in “The Peptides”; Volume 3 (editors: E. Gross and J. Meienhofer), Academic Press, London and New York 1981, in “Methoden der organischen Chemie”, Houben-Weyl, 4^th edition, Vol. 15/1, Georg Thieme Verlag, Stuttgart 1974, in H.-D. Jakubke and H. Jescheit, “Aminosauren, Peptide, Proteine”, Verlag Chemie, Weinheim, Deerfield Beach, and Basel 1982, and/or in Jochen Lehmann, “Chemie der Kohlenhydrate: Monosaccharide and Derivate”, Georg Thieme Verlag, Stuttgart 1974. The protecting groups may be removed at a convenient subsequent stage using methods known from the art.

[00135] The subject compounds can be synthesized via a variety of different synthetic routes using commercially available starting materials and/or starting materials prepared by conventional synthetic methods. A variety of examples of synthetic routes that can be used to synthesize the compounds disclosed herein are described in the schemes below.

Example 1: Preparation of compound 1-2 (stereoismer): (R)-2-((S)-4-(3-amino-6-(2-chloro- lH-pyrrolo[2,3-b]pyridin-3-yl)-5-fluoropyridin-2-yl)piperazin-2-yl)-3-methyIbutan-2-oI

Step 1 tert-butyl (2,5-difluoropyridin-3-yl)carbamate

[00136] To a solution of 3-bromo-2,5-difluoropyridine (2.5 g, 12.89 mmol) in dioxane (30 mL) was added tert-butyl carbamate (1.81 g, 15.47 mmol), CS2CO3 (8.40 g, 25.78 mmol), Xantphos (2.98 g, 5.16 mmol) and Pd(OAc)2 (578.70 mg, 2.58 mmol). The reaction was stirred at 100 °C for 1 h under N?. LCMS showed 26% peak with desired mass. The reaction was cooled to room temperature and filtered through diatomite, the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography silica gel (S i O2 , Petroleum ether/Ethyl acetate = 5/1) to afford tert-butyl (2,5-difluoropyridin-3-yl)carbamate (2.9 g, 97.74% yield) as yellow oil.’H NMR (400 MHz, DMSO-de) 5 9.58 (s, 1H), 8.22-8.08 (m, 1H), 7.87 (t, J = 2.4 Hz, 1H), 1.48 (s, 9H).

Step 2: 6-bromo-2,5-difluoropyridin-3-amine

[00137] To a solution of tert-butyl (2,5-difluoropyridin-3-yl)carbamate (1.5 g, 6.52 mmol) in MeCN (30 mL) and DMF (0.05 mL) was added NBS (1.22 g, 6.84 mmol). The reaction was stirred at 80 °C for 16 h. LCMS showed 75% peak with desired mass. The reaction was cooled to room temperature and poured into saturated N’aHCCL solution (40 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (20 mL x 2) and dried over NaiSO-i, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography silica gel (SiCL, Petroleum ether/Ethyl acetate = 5/1) to afford 6-bromo-2,5 -difluorop yridin-3 -amine (500 mg, 36.72% yield) as a yellow solid. ' H NMR (400 MHz, DMSO-cfo) 5 7.13 (t, J = 8.4 Hz, 1H), 6.02 (s, 2H).

Step 3: (R)-2-((S)-4-(3-amino-6-bromo-5-fluoropyridin-2-yl)piperazin-2-yl)-3-methylbutan-2- ol

[00138] To a solution of (R)-3-methyl-2-((S)-piperazin-2-yl)butan-2-ol (370.93 mg, 2.15 mmol) in NMP (4 mL) was added 6-bromo-2,5-difluoropyridin-3-amine (450 mg, 2.15 mmol) and DIEA (695.71 mg, 5.38 mmol). The reaction was stirred at 120 °C for 32 h. LCMS showed 45% peak with desired mass. The reaction was cooled to room temperature and diluted with H2O (5 mL), and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (5 mL x 2) and dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography silica gel (SiCh, Dichloromethane (0.1% NH3*H2O)/Methanol = 10/1) to afford (R)-2-((S)-4-(3-amino-6-bromo-5-fluoropyridin-2- yl)piperazin-2-yl)-3-methylbutan-2-ol (390 mg, 50.14% yield) as a yellow solid.'H NMR (400 MHz, DMSO-&) 8 6.94 (d, 7 = 9.6 Hz, 1H), 5.31 (s, 2H), 3.98 (s, 1H), 3.28 (d, 7 = 12.0 Hz, 1H), 3.10-2.90 (m, 2H), 2.89-2.70 (m, 2H), 2.45-2.35 (m, 2H), 1.82-1.54 (m, 2H), 1.00 (s, 3H), 0.87- 0.82 (m, 6H).

Step 4: preparation of (R)-2-((S)-4-(3-amino-6-(2-chloro-l-((2-(trimethylsilyl)ethoxy)methyl)- lH-pyrrolo[2,3-b]pyridin-3-yl)-5-fluoropyridin-2-yl)piperaz,in-2-yl)-3-methylbutan-2-ol [00139] To a solution of 2-chloro-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2,3-b]pyridine (407.38 mg, 996.53 pmol) in dioxane (2 mL) and H2O (0.2 mL) was added (R)-2-((S)-4-(3-amino-6-bromo-5-fluoropyridin-2- yl)piperazin-2-yl)-3-methylbutan-2-ol (180 mg, 498.27 mol), K2CO3 (137.73 mg, 996.53 mol) and Pd(dppf)C12 (36.46 mg, 49.83 pmol). The reaction was stirred at 80 °C for 2 h under N2. LCMS showed 64% peak with desired mass. The reaction was cooled to room temperature and diluted with H2O (5 mL). The mixture was extracted with ethyl acetate (20 mL x 3) and the combined organic layers were washed with brine (20 mL x 2) and dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography silica gel (SiCL, Dichloromethane (0.1% NH3*H2O)/Methanol = 5/1) to afford (R)-2-((S)-4-(3-amino-6-(2-chloro-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2,3- b]pyridin-3-yl)-5-fluoropyridin-2-yl)piperazin-2-yl)-3-methylbutan-2-ol (200 mg, 71.27% yield) as a yellow solid. ^]H NMR (400 MHz, DMSO-cfo) 5 8.34 (dd, J = 1.6, 4.8 Hz, 1H), 8.02-7.92 (m, 1H), 7.22 (dd, J = 4.8, 8.0 Hz, 1H), 6.99-6.88 (m, 1H), 5.73 (s, 2H), 5.36 (s, 2H), 4.18-4.02 (m, 1H), 3.64-3.56 (m, 2H), 3.42 (d, J = 11.6 Hz, 1H), 3.23-3.17 (m, 1H), 3.05-2.91 (m, 3H), 2.65- 2.60 (m, 1H), 2.43 (s, 1H), 1.75-1.68 (m, 1H), 1.00 (s, 3H), 0.87-0.79 (m, 8H), -0.10 (s, 9H).

Step 5 (R)-2-((S)-4-(3-amino-6-(2-chloro-lH-pyrrolo[2,3-b]pyridin-3-yl)-5-fluoropyridin-2- yl)piperazin-2-yl)-3-methylbutan-2-ol

[00140] A mixture of (R)-2-((S)-4-(3-amino-6-(2-chloro-l-((2-(trimethylsilyl)ethoxy)methyl)- lH-pyrrolo[2,3-b]pyridin-3-yl)-5-fluoropyridin-2-yl)piperazin-2-yl)-3-methylbutan-2-ol (100 mg, 177.56 pmol) in TBAF (1 M, 5 mL) was stirred at 60 °C for 16 h. LCMS showed 32% peak with desired mass. The reaction was cooled to room temperature and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography silica gel (SiCb, Dichloromethane (0.1 NFL^,H2O (/Methanol = 10/1) to give a crude product. The crude product was purified by prep-HPLC (Neutral conditions) to afford (R)-2-((S)-4-(3-amino-6-(2-chloro-lH- pyrrolo[2,3-b]pyridin-3-yl)-5-fhioropyridin-2-yl)piperazin-2-yl)-3-methylbutan-2-ol (14.1 mg, 18.16% yield) as a white solid.’H NMR (400 MHz, DMSO-afe) 5 8.25 (dd, J = 1.6, 4.8 Hz, 1H), 7.98-7.88 (m, 1H), 7.12 (dd, 7 = 4.8, 8.0 Hz, 1H), 6.92 (d, J = 11.6 Hz, 1H), 5.38-5.14 (m, 2H), 3.95 (s, 1H), 3.38 (d, .J = 11.2 Hz, 1H), 3.15 (d, J = 11.2 Hz, 1H), 3.03-2.95 (m, 1H), 2.94-2.85 (m, 1H), 2.83-2.76 (m, 1H), 2.58-2.54 (m, 1H), 2.47-2.41 (m, 1H), 1.77-1.68 (m, 1H), 1.00 (s, 3H), 0.88-0.82 (m, 6H).

Example 2: Preparation of compound 1-3: (R)-2-((S)-4-(6-(2-chloro-lH-pyrroIo[2,3- b]pyridin-3-yl)-3,5-difluoropyridin-2-yl)piperazin-2-yl)-3-methylbutan-2-ol

Step 1; preparation of (R)-2-((S)-4-(6-bromo-3,5-difhioropyridin-2-yl)piperazin-2-yl)-3- methylbutan-2-ol

[00141] To a solution of 2-bromo-3,5,6-trifluoropyridine (5 g, 23.59 mmol) and (R)-3-methyl- 2-((S)-piperazin-2-yl)butan-2-ol (4.06 g, 23.59 mmol) in EtOH (50 mL) was added DIEA (6. 10 g, 47.18 mmol). The mixture was stirred at 80 °C for 16 h. LCMS showed 86% peak with desired mass. After cooling to room temperature, the reaction mixture was concentrated under reduced pressure to give crude product. The crude product was purified by column chromatography (S i O2, Dichloromethane/Methanol = 10/1) to afford (R)-2-((S)-4-(6-bromo-3,5-difluoropyridin-2- yl)piperazin-2-yl)-3-methylbutan-2-ol (6.2 g, 72.16% yield) as white solid.

Step 2.- preparation of (R)-2-((S)-4-(6-(2-chloro-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrrolo[2,3-b]pyridin-3-yl)-3,5-difluoropyridin-2-yl)piperazin-2-yl)-3-methylbutan-2-ol

[00142] To a mixture of R)-2-((S)-4-(6-bromo-3,5-difluoropyridin-2-yl)piperazin-2-yl)-3- methylbutan-2-ol (6.2 g, 17.02 mmol), 2-chloro-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)- l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2,3-b]pyridine (6.96 g, 17.02 mmol) in dioxane (70 mL) and H₂O (7 mL) was added Pd(dppf)Cl₂ (1.25 g, 1.70 mmol), K₂CO₃ (7.06 g, 51.07 mmol). The mixture was stirred at 80 °C for 16 h under N₂. LCMS showed 48% peak with desired mass. After cooling to room temperature, the reaction mixture was poured into water (50 mL), extracted with ethyl acetate (30 mL x 2). The combined organic layers were washed with brine (40 mL), dried over Na₂SO4, filtered and concentrated under reduced pressure to give crude product. The crude product was purified by column chromatography (SiO₂, Dichloromethane/Methanol = 10/1) to afford (R)-2-((S)-4-(6-(2-chloro-l-((2-

(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2,3-b]pyridin-3-yl)-3,5-difluoropyridin-2- yl)piperazin-2-yl)-3-methylbutan-2-ol (6.6 g, 68.48% yield) as brown oil.

Step 3: preparation of (R)-2-((S)-4-(6-(2-chloro-lH-pyrrolo[2,3-bJpyridin-3-yl)-3,5- difluoropyridin-2-yl)piperazin-2-yl)-3-methylbutan-2-ol [00143] To a solution of (R)-2-((S)-4-(6-(2-chloro-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrrolo[2,3-b]pyridin-3-yl)-3,5-difluoropyridin-2-yl)piperazin-2-yl)-3-methylbutan-2-ol (6.6 g, 11.66 mmol) in DCM (66 mL) was added TFA (66 mL). The mixture was stirred at 20 °C for 2 h. LCMS showed 90% peak with desired mass. The reaction mixture was concentrated under reduced pressure to give crude product. The crude product was purified by prep-HPLC (FA conditions) to afford (R)-2-((S)-4-(6-(2-chloro-lH-pyrrolo|2,3-b|pyridin-3-yl)-3,5- difluoropyridin-2-yl)piperazin-2-yl)-3-methylbutan-2-ol (949.1 mg, FA salt, 16.64% yield) as a white solid.’H NMR (400 MHz, DMSO-tfo) 5 8.29 (dd, J = 1.6, 4.8 Hz, 1H), 8.22 (s, 1H), 7.97- 7.93 (m, 1H), 7.91-7.88 (m, 1H), 7.18-7.15 (m, 1H), 4.54-4.20 (m, 1H), 4.05 (d, J = 12.0 Hz, 1H), 3.78 (d, J = 8.8 Hz, 1H), 3.07-3.05 (m, 1H), 2.88-2.82 (m, 3H), 2.77-2.74 (m, 1H), 1.78-1.75 (m, 1H), 1.02 (s, 3H), 0.86 (d, J = 6.4 Hz, 3H), 0.76 (d, J = 6.8 Hz, 3H).

Example 3: Cell Based Assay

[00144] 1) Construction of Jurkat stable cell line.

Jurkat, clone E6-1 cells (TIB- 152) were purchased from ATCC (ATCC®, Manassas, VA, USA). pNL3.2.NF-kB-RE Vector were purchased from (GenBank® Accession Number: JQ513377. Promega®, Wisconsin, USA). ViaFect™ Transfection Reagent was used to transfect Jurkat cells with the plasmids. Transfected cells were maintained in RPMI-1640 supplemented with 10% fetal bovine serum, 100 units/mL Penicillin G, 100 pg/mL Streptomycin, and 400 pg/mL Hygromycin B Solution. The best cell line was identified based on the reporter assay results. When clone selection, each cell line was stimulated by TNF-alfa instead of anti CD3 antibody and anti CD28 antibody. The best cell line #1450 pNL3.2NFkB-RE/Jurkat clone No.29 was cryopreserved in CELL BANKER1 plus.

[00145] 2) Preparation of anti CD3 antibody coated plate.

The 384 well assay plates which were applied with 5 pg/mL Anti CD3 antibody diluted with D-PBS(-) or D-PBS(-) were incubated overnight or 3 days at 4 °C prior to two times of washing with 50 pL PBS by upside-down centrifuge for 10 sec. Maximum speed was 1000 rpm. [00146] 3) Cell plating and stimulating with anti CD3 antibody, anti CD28 antibody, and test drugs.

The frozen #1450 pNL3.2NFkB-RE/Jurkat clone No.29 in CELLBANKER1 plus (1 ml) was thawed at 37 °C and diluted with 9 mL RPMI-1640 medium supplemented with 10% FBS and 100 units/mL Penicillin G, 100 pg/mL Streptomycin (Assay medium). Cells were centrifuged for 3 min at 1 ,000 rpm and supernatant was removed prior to resuspending cells in 5mL of the assay medium and counting cells. 20 pL prepared 1 x 106 cells/mL cell suspension (20000 cells/well) or 20 pL assay medium were plated out onto anti CD3 antibody coated 384 plates. Subsequently, 2.5 pL test drugs containing 1% Dimethyl sulfoxide (DMSO) and 2.5 qL 50 qg/mL anti CD28 solution diluted with assay medium were added to each well. The plates were incubated overnight at 37 °C. in a humidified atmosphere containing 5% CO2. All test drugs were dissolved in DMSO as a stock solution and diluted with assay medium. Final DMSO concentration for assay samples was 0.1%.

[00147] 4) Cell viability assay: measurement of ATP.

Assay plates (3) were incubated for 30 min at room temperature prior to addition of 12.5 pL Cell titer- Gio, mixed for 3 min at 900 rpm and centrifuged for 10 sec at 1000 rpm. Luminescence was recorded using EnVision® luminescence recorder (PerkinElmer).

[00148] 5) Reporter assay: Measurement of NF-kappaB.

Assay plates (3) were incubated for 30 min at room temperature prior to addition of 12.5 pL x Nano-Gio Luciferase Assay Substrate diluted with Nano-Gio Luciferase Assay Buffer, mixed for 3 min at 900 rpm and centrifuged for 10 sec at 1000 rpm. Luminescence was read using EnVision® (PerkinElmer).

[00149] 6) The plate map is shown in FIG. 1.

[00150] 7) Data Analysis of Cell Viability assay

Data was analyzed using XLfit 5.5.0.5, (ID Business Solutions Limited., Boston, MA, USA).

[00151] % inhibition = [[-(Sample - Low Control)/ (High control - Low Control)] x 100 [00152] Low control: Average RLU of lane 24

[00153] High control: Average RLU of lane 22

[00154] Dose response curve was analyzed by 4 Parameter Logistic Model and IC50 was calculated as y=50.

[00155] 4 Parameter Logistic Model

[00156] fit = (A+((B-A)/(l+(10^A((C-x)*D)))))

[00157] inv = (C-(log(((B-A)/(y-A))-l)/D))

[00158] res = (y-fit)

[00159] 8) Data Analysis of Reporter assay

Data was analyzed using XLfit 5.5.0.5,

[00160] % inhibition = [[-(Sample - Low Control)/ (High control - Low Control)] x 100

[00161] Low control: Average RLU of lane 23

[00162] High control: Average RLU of lane 22

[00163] Dose response curve was analyzed by 4 Parameter Logistic Model and IC50 was calculated as y=50.

[00164] 4 Parameter Logistic Model [00165] fit = (A+((B-A)/(l+(10^A((C-x)*D)))))

[00166] inv = (C-(log(((B-A)/(y-A))-l)/D))

[00167] res = (y-fit)

Example 4: Enzyme Assay

[00168] 1) Enzyme assay

Each concentration of enzyme with 0.5x PKC LIPID activator solution in assay buffer (2 pL/well) were added to the 384-well plate (greiner # 784075) (See table 1 below for prepared and final concentrations).

[00169] Table 1:

[00170] The compound diluted in the assay buffer (2 pL/well ) was added to the 384-well plate. Mixture of enzyme and compound was incubated for 60 minutes at room temperature. Each prepared concentration of ULight-CREBtide (Serl33) and ATP diluted in assay buffer (2 pL/well) were added to the 384-well plate. (See table 2 below for prepared and final concentrations).

[00171] Table 2:

[00172] Mixture of enzyme, compound, ULight reagent and ATP were incubated for 60 minutes at room temperature. 0.5 nM Eu-anti-p-CREB and 40 mM EDTA diluted in IX LANCE detection Buffer (6 pL/well) were added to the 384-well plate. Mixtures are incubated for 60 minutes at room temperature. TR-FRET signal was measured with Envision (PerkinElmer).

[00173] A665 = Emission at 665 nm (RFU)D615 = Emission at 615 nm (RFU)RFU : Relative Fluorescence UnitsRatio=A665/D615 10000

[00174] 2) Plate map is shown in FIG. 2. [00175] 3) Data Analysis of Enzyme assay

Data were analyzed using XLfit 5.5.0.5, (ID Business Solutions Limited., Boston, MA, USA).

[00176] % inhibition = [l-(Sample - Low Control)/ (High control - Low Control)] x 100

[00177] Low control: Average RLU of lane 24 (each well without enzymes)

[00178] High control: Average RLU of lane 22 and 23 (each well with enzymes)

[00179] Dose response curve was analyzed by 4 Parameter Logistic Model and IC50 was calculated as y=50.

[00180] 4 Parameter Logistic Model

[00181] fit = (A+((B-A)/(l+(10^A((C-x)*D)))))

[00182] inv = (C-(log(((B-A)/(y-A))-l)/D))

[00183] res = (y-fit)

Results

[00184] The resulting IC50 data from the cell based and enzyme assays are shown in Table 3 below. The data was collected for compounds described herein and is noted in relation to the particular type of PKC tested, i.e., theta, betal, delta, or epsilon. The legend for the table is: “A” for < 1 nM, “B” for between 1 nM and 10 nM, “C” for between 10 nM and 100 nM, “D” for 100 nM or greater, and a blank space when the particular data point was not recorded.

[00185] Table 3:

Example 5: PKC theta inhibitors in sRICA: Stimulated with anti-CD3 and anti-CD28

[00186] FIG. 3 and 4 show sRICA assays. The resulting IC50 data from the sRICA assays are shown in Table 4 below.

[00187] Table 4:

Example 6: In vivo mouse data

Materials

Animals

[00188] Female Balb/c mice, weighing 18-21 grams and 8 weeks old (purchased from Jihui), were acclimated for approximately 5 days before the start of the experiment. Five mice were housed per cage, maintained at a temperature of 24°C ± 1°C and a humidity of 55% ± 5%. A light/dark cycle of 12 hours was followed (lights on at 8:00 AM and off at 8:00 PM). Mice had free access to food and water during this period.

Main Reagents/Supplies

Experimental Methods and Procedures

[00189] 1. Accurately weighed an appropriate amount of EDTA K2 powder and dissolved it in deionized water to prepare a 150 mg/mL anticoagulant solution. Stored at 4°C for later use. Before the experiment, aliquoted 3 pL into each 1.5 mL centrifuge tube to serve as anticoagulant tubes.

[00190] 2. Before the experiment, an appropriate amount of InVivoMAb anti-mouse CD3e antibody was taken and diluted it to a concentration of 50 pg/mL using InVivoPure pH 7.0 Dilution Buffer. Placed on wet ice until ready for use.

[00191] 3. Prepared the test compounds at the required concentration using the following formation: 10% DMSO, 50% PEG400, 40% aqueous solution containing 20% HP- -CD (pH=4.95).

[00192] 4. Recorded the body weight of all mice and randomly assigned them into control and experimental groups, with 6 mice in each group. [00193] 5. Administered the test compound to the mice in the experimental group via oral gavage, while the control group receives the corresponding solvent as a control.

[00194] 6. One hour after administration, injected all mice intraperitoneally with 100 pL of the prepared anti-CD3e antibody solution (5 pg per mouse).

[00195] 7. At the time points of 1, 3, and 8 hours after the injection of anti-CD3e antibody, collected blood from the orbital sinus of each mouse and transferred it into EDTA K2 anticoagulant tubes. Centrifuged at 3000 RCF for 5 minutes at 4°C to obtain the plasma supernatant. During the experiment, handled the samples on ice, and if not analyzed immediately, stored them at -80°C to avoid repeated freeze-thaw cycles.

[00196] 8. Measured the plasma concentration of IL-2 using a mouse IL-2 ELISA kit (Solabio, SEKM-0004). Followed the operating procedures according to the technical instructions provided by the supplier. Thirty minutes prior to the experiment, the kit was removed from storage and was allowed to reach room temperature. Soaked the microtiter plates three times and then dried them. Added 100 pL of standard and detection samples to the reaction wells, sealed the plate, and incubated at 37°C for 90 minutes. Washed the plate four times and dried it thoroughly. Next, added 100 pL of the biotinylated antibody working solution to each reaction well, sealed the plate, and incubated at 37°C for 60 minutes. Washed the plate four times and dried it completely. Subsequently, added 100 pL of the enzyme conjugate working solution to the reaction wells, sealed the plate, and incubated at 37°C for 30 minutes. Washed the plate five times and dried it well. Finally, added 100 pL of chromogenic substrate to each reaction well, allowing the color to develop at 37°C for approximately 15 minutes. Added 50 pL of stop solution and promptly measured the OD values using Ensighf™ Microplate reader (Revvity)at a wavelength of 450 nm within five minutes.

[00197] 9. Data analysis was conducted using GraphPad version 9.4.1, employing one-way ANOVA as the analytical method.

[00198] FIG. 5 to FIG. 10 show in vivo mouse data. The results in mice are shown in Table 5 below.

[00199] Table 5:

[00200] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it is readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

[00201] Accordingly, the preceding merely illustrates the principles of the invention. It will be appreciated that those skilled in the art will be able to devise various arrangements which, although not explicitly described or shown herein, embody the principles of the invention and are included within its spirit and scope. Furthermore, all examples and conditional language recited herein are principally intended to aid the reader in understanding the principles of the invention and the concepts contributed by the inventors to furthering the art, and are to be construed as being without limitation to such specifically recited examples and conditions. Moreover, all statements herein reciting principles, aspects, and embodiments of the invention as well as specific examples thereof, are intended to encompass both structural and functional equivalents thereof. Additionally, it is intended that such equivalents include both currently known equivalents and equivalents developed in the future, i.e., any elements developed that perform the same function, regardless of structure. Moreover, nothing disclosed herein is intended to be dedicated to the public regardless of whether such disclosure is explicitly recited in the claims.

[00202] The scope of the present invention, therefore, is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of present invention is embodied by the appended claims. In the claims, 35 U.S.C. §112(f) or 35 U.S.C. §112(6) is expressly defined as being invoked for a limitation in the claim only when the exact phrase "means for" or the exact phrase "step for" is recited at the beginning of such limitation in the claim; if such exact phrase is not used in a limitation in the claim, then 35 U.S.C. § 112 (f) or 35 U.S.C. §112(6) is not invoked.

Claims

What Is Claimed Is:

A compound of formula (I):

wherein:

Z is CR³ or N;

X is a bicyclic and optionally substituted heteroaryl group;

R¹ and R² are each independently selected from the group consisting of H, alkyl, halo, amine, cyano, and hydroxyl, or R¹ and R² together with the atoms to which they are attached form a 5-membered ring or a 6-membered aryl ring or a heteroaryl ring;

R³ is selected from the group consisting of H and halo; and

R⁴ and R⁵ are each independently selected from the group consisting of H, C2-4 linear alkyl, C2-4 branched alkyl and C3-6 cycloalkyl group.

2. The compound of claim 1, wherein the compound has formula (la):

wherein:

Z is CR³ or N;

X is a bicyclic and optionally substituted heteroaryl group;

R¹ is -CH-;

R² is -NH-;

R³ is selected from the group consisting of H and halo; and

R⁴ and R⁵ are each independently selected from the group consisting of H, C2-4 linear alkyl, C2-4 branched alkyl and C3-6 cycloalkyl group.

3. The compound of claim 1, wherein the compound has formula (lb):

wherein:

Z is CR³;

X is a bicyclic and optionally substituted heteroaryl group;

R¹ is -NH-, -S-, -O-, or -N-lower alkyl;

R² is -CH-, -NH-, -S-, -O-, or -N-lower alkyl;

R³ is selected from the group consisting of H and halo; and

R⁴ and R⁵ are each independently selected from the group consisting of H, C2-4 linear alkyl, C2-4 branched alkyl and C3-6 cycloalkyl group.

4. The compound of claims 1-2, wherein Z is CR³.

5. The compound of claim 4, wherein R³ is F, Cl, Br, or I.

6. The compound of claim 5, wherein R³ is F.

7. The compound of claims 1-2, wherein Z is N.

8. The compound of claims 1-3, wherein X has the structure:

wherein: ring A is a 5-membered and optionally substituted heteroaryl group; and ring B is a 6-membered and optionally substituted heteroaryl group.

9. The compound of claim 8, wherein ring A is selected from the group consisting of:

10. The compound of any one of claims 8-9, wherein ring B is selected from the group consisting of pyridine, and substituted versions thereof.

11. The compound of claims 1-3, wherein R¹ is F, Cl, Br, I, or NH2.

12. The compound of claim 11, wherein R¹ is NH2.

13. The compound of claim 11, wherein R¹ is Cl.

14. The compound of claim 11, wherein R¹ is F.

15. The compound of any one of claims 1-3, wherein R² is H.

16. The compound of any one of claims 1-3, wherein R³ is H, F, Cl, or Br.

17. The compound of claim 16, wherein R³ is F.

18. The compound of claim 16, wherein R³ is H.

19. The compound of claim 1, wherein R¹ is Cl, R² is H, and R³ is F.

20. The compound of claim 1, wherein R¹ is NH2, R² is H, and R is F.

21. The compound of claim 1, wherein R¹ is F, R² is H, and R³ is F.

22. The compound of claim 2, wherein Z is C; R¹ is -CH-, R² is -NH-; and R³ is halo.

23. The compound of claim 2, wherein Z is C; R¹ is -NH-, R² is -CH-; and R³ is halo.

24. The compound of claim 23, wherein R³ is F.

25. The compound of claim 3, wherein Z is N; R¹ is -CH-, R² is -NH-; and R³ is H.

26. The compound of claims 1-3, wherein R⁴ is isopropyl or tertbutyl.

27. The compound of claim 26, wherein R⁴ is isopropyl.

28. The compound of claims 1-3, wherein the compound has formula (Ic):

29. The compound of claims 1-3, wherein the compound has formula (Id):

30. The compound of claims 1-3, wherein the compound has formula (le):

(le).

31. A compound selected from:

1-4

32. A pharmaceutical composition comprising a compound according to any one of claims 1-31.

33. A method of treating a subject for a condition, the method comprising administering to the subject a compound of any one of claims 1-32 or a pharmaceutical composition according to claim 32.

34. A method of treating a subject for inflammatory bowel disease, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-32 or a pharmaceutical composition according to claim 32.

35. The method of claim 34, wherein the inflammatory bowel disease is Crohn’s disease or ulcerative colitis.