HPK1 antagonists and uses thereof

Abstract

The present invention provides compounds, compositions thereof, and methods of their use for the inhibition of HPK1 and the treatment of HPK1-mediated disorders. The compounds of the present invention and their pharma- ceutically acceptable compositions are useful for the treatment of various diseases, disorders, or conditions associated with the regulation of signal transduction pathways involving HPK1 kinase. Such diseases, disorders, or conditions include those described herein. The compounds provided by the present invention are also useful for the study of HPK1 enzyme in biological and pathological phenomena, the study of intracellular signal transduction pathways occurring in body tissues, and the comparative evaluation of new HPK1 inhibitors or other regulators of kinases, signal transduction pathways, and cytokine levels in vitro or in vivo.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 63 / 157,230, filed March 5, 2021, the entire contents of which are incorporated herein by reference.

[0002] The present invention relates to compounds and methods useful for antagonizing hematopoietic progenitor kinase 1 (HPK1). The present invention also provides pharmaceutically acceptable compositions comprising the compounds of the invention, and methods of using the compositions in the treatment of various disorders. [Background technology]

[0003] Hematopoietic progenitor kinase 1 (HPK1), also known as mitogen-activated protein kinase kinase kinase kinase 1 (MAP4K1), is a hematopoietic cell-restricted member of the Ste20 serine / threonine kinase superfamily. The MAP4K family includes MAP4K1 / HPK1, MAP4K2 / GCK, MAP4K3 / GLK, MAP4K4 / HGK, MAP4K5 / KHS, and MAP4K6 / MINK. HPK1 is a tissue-specific upstream activator of the MEKK / JNK / SAPK signaling pathway.

[0004] HPK1 is of particular interest because it is expressed primarily in hematopoietic cells such as T cells, B cells, macrophages, dendritic cells, neutrophils, and mast cells (Hu, MC, et al., Genes Dev, 1996.10(18):pp.2251-64; Kiefer, F., et al., EMBO J, 1996.15(24):pp.7013-25). HPK1 kinase activity has been shown to be induced upon activation of the T cell receptor (TCR) (Liou, J., et al., Immunity, 2000.12(4):pp.399-408), B cell receptor (BCR) (Liou, J., et al., Immunity, 2000.12(4):pp.399-408), transforming growth factor receptor (TGF-PR) (Wang, W., et al., J Biol Chem, 1997.272(36):pp.22771-5; Zhou, G., et al., J Biol Chem, 1999.274(19):pp.13133-8), or Gs-coupled PGE2 receptors (EP2 and EP4) (Ikegami, R., et al., J Immunol, 2001.166(7):pp.4689-96). Thus, HPK1 regulates diverse functions of various immune cells. HPK1 is an example of a negative regulator of dendritic cell activation and T and B cell responses that can be targeted to enhance antitumor immunity. HPK1 is primarily expressed by hematopoietic cells, including early progenitor cells. In T cells, HPK1 is thought to negatively regulate T cell activation by phosphorylating SLP76 at Ser376 (Di Bartolo et al. (2007) JEM 204:681-691) and Gads at Thr254, thereby reducing the persistence of signaling microclusters, leading to the recruitment of 14-3-3 proteins that bind to phosphorylated SLP76 and Gads, resulting in the release of the SLP76-Gads-14-3-3 complex from LAT-containing microclusters (Lasserre et al. (2011) J Cell Biol 195(5):839-853). HPK1 is also activated in response to prostaglandin E2, which is often secreted by tumors, and may contribute to tumor cell evasion from the immune system. HPK1 is important in regulating the functions of various immune cells and is involved in autoimmune diseases and anti-tumor immunity (Shui, JW, et al., Nat Immunol, 2007.8(1):pp.84-91; Wang, X., et al., J Biol Chem, 2012.287(14):pp.11037-48). [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Hu, MC, et al., Genes Dev, 1996.10(18):p.2251-64 [Non-patent document 2] Kiefer, F., et al., EMBO J, 1996.15(24):p.7013-25 [Non-patent document 3] Liou, J., et al., Immunity, 2000.12(4):p.399-408 [Non-patent document 4] Wang, W., et al., J Biol Chem, 1997.272(36):p.22771-5 [Non-Patent Document 5] Zhou, G., et al., J Biol Chem, 1999.274(19):p.13133-8 [Non-patent document 6] Ikegami, R., et al., J Immunol, 2001.166(7):p.4689-96 [Non-Patent Document 7] Di Bartolo et al. (2007) JEM 204:681-691 [Non-patent document 8] Lasserre et al. (2011) J Cell Biol 195(5):839-853 Summary of the Invention [Means for solving the problem]

[0006] It has now been found that compounds of the present invention, and pharmaceutically acceptable compositions thereof, are effective as antagonists of HPK1. In certain embodiments, the present invention provides compounds of the formulae presented herein.

[0007] The compounds of the present invention, and pharmaceutically acceptable compositions thereof, are useful for treating a variety of diseases, disorders, or conditions associated with modulation of signal transduction pathways involving HPK1 kinase, including those described herein.

[0008] The compounds provided by the present invention are also useful for studying the HPK1 enzyme in biological and pathological phenomena, for studying intracellular signaling pathways occurring in body tissues, and for the comparative evaluation of new HPK1 inhibitors or other regulators of kinases, signaling pathways, and cytokine levels in vitro or in vivo. DETAILED DESCRIPTION OF THE INVENTION

[0009] Detailed Description of Specific Embodiments 1. Overview of Certain Embodiments of the Invention: In certain embodiments, the present invention provides a compound of formula I [ka] or a pharmaceutically acceptable salt thereof, wherein X, Z, R 1 , R 2 , R 3 and R 1 and R 2 are each, both alone and in combination, as defined below and as described in embodiments herein, or a pharmaceutically acceptable salt thereof.

[0010] In some embodiments, the present invention provides pharmaceutical compositions comprising a compound of formula I and a pharmaceutically acceptable carrier, adjuvant, or diluent.

[0011] In some embodiments, the present invention provides a method of treating an HPK1-mediated disease, disorder, or condition, comprising administering to a patient in need thereof a compound of formula I, or a pharmaceutically acceptable salt thereof.

[0012] 2. Compounds and Definitions: Compounds of the present invention include those generally described herein and are further designated by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. For purposes of the present invention, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Edition. Furthermore, the general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5th Edition. th Ed., Ed.: Smith, MB and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are incorporated herein by reference.

[0013] The terms "aliphatic" or "aliphatic group," as used herein, mean a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more units of unsaturation, having a single point of attachment to the rest of the molecule; or a monocyclic or bicyclic hydrocarbon that is fully saturated or contains one or more units of unsaturation, but is not aromatic (also referred to herein as "carbocyclic," "alicyclic," or "cycloalkyl"). Unless otherwise specified, an aliphatic group contains 1-6 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-5 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-4 aliphatic carbon atoms. In yet other embodiments, an aliphatic group contains 1-3 aliphatic carbon atoms, and in yet other embodiments, an aliphatic group contains 1-2 aliphatic carbon atoms. In some embodiments, "alicyclic" (or "carbocycle" or "cycloalkyl") refers to a monocyclic C3-C6 hydrocarbon that is fully saturated or contains one or more units of unsaturation, but is not aromatic, having a single point of attachment to the rest of the molecule. Suitable aliphatic groups include, but are not limited to, straight-chain or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups, and hybrids thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl.

[0014] As used herein, the term "bridged bicyclic" refers to any bicyclic ring system, i.e., carbocyclic or heterocyclic, saturated or partially unsaturated, having at least one bridge. As defined by IUPAC, a "bridge" is an unbranched chain of atoms or a valence bond connecting two bridgehead positions, where a "bridgehead" is any skeletal atom of the ring system that is bonded to three or more skeletal atoms (excluding hydrogen). In some embodiments, the bridged bicyclic group has 7 to 12 ring members and 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Such bridged bicyclic groups are well known in the art and include the groups shown below, in which each group is attached to the remainder of the molecule at any substitutable carbon or nitrogen atom. Unless otherwise specified, bridged bicyclic groups are optionally substituted with one or more substituents as shown for aliphatic groups. Additionally or alternatively, any substitutable nitrogen of the bridged bicyclic group is optionally substituted. Exemplary bridged bicyclic groups include: [ka] [ka]

[0015] The term "lower alkyl" refers to C 1-4 It refers to a straight-chain or branched alkyl group. Exemplary lower alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and tert-butyl.

[0016] The term "lower haloalkyl" refers to a C alkyl group substituted with one or more halogen atoms. 1-4 It refers to a straight or branched chain alkyl group.

[0017] The term "heteroatom" refers to oxygen, sulfur, nitrogen, phosphorus, or silicon (any oxidized form of nitrogen, sulfur, phosphorus, or silicon, the quaternized form of any basic nitrogen, or a substitutable nitrogen of a heterocycle, for example, N (such as 3,4-dihydro-2H-pyrrolyl), NH (such as pyrrolidinyl), or NR +(including N-substituted pyrrolidinyl, etc.)

[0018] As used herein, the term "unsaturated" means that a moiety has one or more units of unsaturation.

[0019] As used herein, "divalent C 1-8 (or C 1-6 The term "saturated or unsaturated, straight or branched, hydrocarbon chain" refers to divalent alkylene, alkenylene, and alkynylene chains that are straight or branched as defined herein.

[0020] The term "alkylene" refers to a divalent alkyl group. An "alkylene chain" is a polymethylene group, i.e., -(CH2) n -, where n is preferably a positive integer of 1 to 6, 1 to 4, 1 to 3, 1 to 2, or 2 to 3. A substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms have been replaced with a substituent. Suitable substituents include those described below for substituted aliphatic groups.

[0021] The term "alkenylene" refers to a divalent alkenyl group. A substituted alkenylene chain is a polymethylene group containing at least one double bond in which one or more methylene hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for substituted aliphatic groups.

[0022] The term "halogen" means F, Cl, Br, or I.

[0023] The term "aryl," used alone or as part of a larger moiety such as "aralkyl," "aralkoxy," or "aryloxyalkyl," refers to a monocyclic or bicyclic ring system having a total of 5 to 14 ring members, wherein at least one ring of the system is aromatic and each ring of the system contains 3 to 7 ring members. The term "aryl" may be used interchangeably with the term "aryl ring." In certain embodiments of the present invention, "aryl" refers to an aromatic ring system, including, but not limited to, phenyl, biphenyl, naphthyl, anthracyl, and the like, which may bear one or more substituents. Also included within the scope of the term "aryl," as used herein, are groups in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthymidyl, phenanthridinyl, or tetrahydronaphthyl.

[0024] The terms "heteroaryl" and "heteroa," used alone or as part of a larger moiety, such as "heteroaralkyl" or "heteroaralkoxy," refer to groups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms, and having 6, 10, or 14 pi-electrons shared in the cyclic array, and having 1 to 5 heteroatoms in addition to carbon atoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur and any quaternized form of a basic nitrogen. Heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. As used herein, the terms "heteroaryl" and "heteroaromatic" also include groups in which a heteroaromatic ring is fused to one or more aryl, alicyclic, or heterocyclyl rings, and unless otherwise specified, the radical or point of attachment is on the heterocyclic ring or on one of the rings to which the heterocyclic ring is fused. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazole, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, and tetrahydroisoquinolinyl. Heteroaryl groups may be monocyclic or bicyclic. The term "heteroaryl" may be used interchangeably with "heteroaryl ring," "heteroaryl group," or "heteroaromatic," any of which terms include optionally substituted rings. The term "heteroaralkyl" refers to an alkyl group substituted by a heteroaryl, wherein the alkyl and heteroaryl portions independently are optionally substituted.

[0025] As used herein, the terms "heterocycle," "heterocyclyl," "heterocyclic radical," and "heterocycle" are used interchangeably and refer to a stable 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated and that, in addition to carbon atoms, has one or more, preferably 1 to 4, heteroatoms as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes substituted nitrogen. By way of example, in a saturated or partially unsaturated ring having 0 to 3 heteroatoms selected from oxygen, sulfur, or nitrogen, the nitrogen can be N (e.g., 3,4-dihydro-2H-pyrrolyl), NH (e.g., pyrrolidinyl), or + It can be NR (such as N-substituted pyrrolidinyl).

[0026] A heterocycle may be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure, and any of the ring atoms may be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenylpyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, 2-oxa-6-azaspiro[3.3]heptane, and quinuclidinyl. The terms "heterocycle," "heterocyclyl," "heterocyclyl ring," "heterocyclic group," "heterocyclic moiety," and "heterocyclic radical" are used interchangeably herein and include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or alicyclic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl. Heterocyclyl groups may be monocyclic or bicyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.

[0027] As used herein, the term "partially unsaturated" refers to a ring moiety that contains at least one double or triple bond. The term "partially unsaturated" is intended to encompass rings with multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties as defined herein.

[0028] As described herein, compounds of the invention may contain "optionally substituted" moieties. In general, the term "substituted," whether preceded by the term "optionally" or not, means that one or more hydrogens of the specified moiety are replaced with a suitable substituent. Unless otherwise indicated, an "optionally substituted group" may have a suitable substituent at each substitutable position of the group, and if more than one position in any given structure can be substituted with more than one substituent selected from a specified group, the substituents may be the same or different at all positions. Combinations of substituents envisioned by the present invention are preferably those that result in the formation of stable or chemically viable compounds. As used herein, the term "stable" refers to a compound that is substantially unchanged when subjected to conditions that allow for its production, detection, and, in certain embodiments, its recovery, purification, and use for one or more of the purposes disclosed herein.

[0029] Suitable monovalent substituents on a substitutable carbon atom of an "optionally substituted" group are independently: halogen; -(CH) 0-4 R;-(CH2) 0-4 OR;-O(CH2) 0-4 R o , -O-(CH2) 0-4 C(O)OR°;-(CH2) 0-4 CH(OR)2;-(CH2) 0-4 SR;-(CH2) 0-4 Ph (which may be substituted by R°); -(CH2) 0-4 O(CH2) 0-1 Ph (which may be substituted with R°); -CH=CHPh (which may be substituted with R°); -(CH2) 0-4 O(CH2) 0-1-pyridyl (which may be substituted by R°); -NO2; -CN; -N3; ​​-(CH2) 0-4 N(R)2;-(CH2) 0-4 N(R)C(O)R;-N(R)C(S)R;-(CH2) 0-4 N(R)C(O)NR2;-N(R)C(S)NR2;-(CH2) 0-4 N(R)C(O)OR;-N(R)N(R)C(O)R;-N(R)N(R)C(O)NR2;-N(R)N(R)C(O)OR;-N(R)C(NR)N(R) 2; -(CH2) 0-4 C(O)R;-C(S)R;-(CH2) 0-4 C(O)OR;-(CH2) 0-4 C(O)SR;-(CH2) 0-4 C(O)OSiR3;-(CH2) 0-4 OC(O)R;-OC(O)(CH2) 0-4 SR°;-SC(S)SR°;-(CH2) 0-4 SC(O)R;-(CH2) 0-4 C(O)NR2;-C(S)NR2;-C(S)SR°;-SC(S)SR°,-(CH2) 0-4 OC(O)NR2;-C(O)N(OR)R;-C(O)C(O)R;-C(O)CH2C(O)R;-C(NOR)R;-(CH2) 0-4 SSR;-(CH2) 0-4 S(O)R;-(CH) 0-4 S(O)2OR;-(CH2) 0-4 OS(O)2R;-S(O)2NR2;-(CH2) 0-4 S(O)R;-N(R)S(O)2NR2;-N(R)S(O)2R;-N(OR)R;-C(NH)NR2;-P(O)2R;-P(O)R2;-OP(O)R2;-OP(O)(OR)2;-SiR3;-(C 1-4 linear or branched alkylene)ON(R)2; or -(C 1-4 linear or branched alkylene)C(O)ON(R)2, where each R may be substituted as defined below and is independently hydrogen, C 1-6 Aliphatic, -CH2Ph, -O(CH2) 0-1Ph, -CH2- (a 5-6 membered heteroaryl ring), or a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or as defined above, but two independent occurrences of R taken together with their intervening atoms form a 3-12 membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which can be substituted as defined below.

[0030] Suitable monovalent substituents on R (or the ring formed by two independent occurrences of R taken together with their intervening atoms) are independently halogen, —(CH) 0-2 R λ ,-(Halo R λ ), -(CH2) 0-2 OH, -(CH2) 0-2 OR λ , -(CH2) 0-2 CH(OR λ )2;-O(HaloR λ ), -CN, -N3, -(CH2) 0-2 C(O)R λ , -(CH2) 0-2 C(O)OH, -(CH2) 0-2 C(O)OR λ , -(CH2) 0-2 SR λ , -(CH2) 0-2 SH, -(CH2) 0-2 NH2, -(CH2) 0-2 NHR λ , -(CH2) 0-2 NR λ 2, -NO2, -SiR λ 3. -OSiR λ 3. -C(O)SR λ 、 -(C 1-4 Linear or branched alkylene)C(O)OR λ , or -SSR λ and each R λ is unsubstituted or, if preceded by "halo", substituted with one or more halogens only, and independently represents C1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 or a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on a saturated carbon atom of R include =0 and =S.

[0031] Preferred divalent substituents on a saturated carbon atom of an "optionally substituted" group are ═O, ═S, ═NNR * 2, =NNHC(O)R * , =NNHC(O)OR * , =NNHS(O)2R * , =NR * , =NOR * , -O(C(R * 2)) 2-3 O- or -S(C(R * 2)) 2-3 Contains S-, R * Each independent occurrence of may be substituted as defined below with hydrogen, C 1-6 A preferred divalent substituent attached to adjacent substitutable carbon atoms of an "optionally substituted" group is selected from aliphatic or unsubstituted 5-6 membered saturated, partially unsaturated, or aryl rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. * 2) 2-3 Contains O- and R * Each independent occurrence of may be substituted as defined below with hydrogen, C 1-6 It is selected from aliphatic or unsubstituted 5-6 membered saturated, partially unsaturated, or aryl rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0032] R * Suitable substituents on the aliphatic groups are halogen, -R λ ,-(Halo R λ ), -OH, -OR λ , -O(HaloR λ ), -CN, -C(O)OH, -C(O)OR λ , -NH2, -NHR λ, -NR λ 2, or -NO2, and each R λ is unsubstituted or, if preceded by "halo", substituted with one or more halogens only, and independently represents C 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, or a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0033] A preferred substituent on a substitutable nitrogen of an "optionally substituted" group is -R † , -NR † 2. -C(O)R † , -C(O)OR † , -C(O)C(O)R † , -C(O)CHC(O)R † , -S(O)2R † , -S(O)NR † 2. -C(S)NR † 2. -C(NH)NR † 2, or -N(R † )S(O)2R † Including each R † are independently halogen, C which may be substituted as defined below 1-6 an aliphatic, unsubstituted -OPh, or an unsubstituted 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or as defined above but containing two independent R † together with their intervening atoms form an unsubstituted 3-12 membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0034] R † Suitable substituents on the aliphatic group are independently halogen, —R λ ,-(Halo R λ ), -OH, -OR λ , -O(HaloR λ ), -CN, -C(O)OH, -C(O)ORλ , -NH2, -NHR λ , -NR λ 2, or -NO2, and each R λ is unsubstituted or, if preceded by "halo", substituted with one or more halogens only, and independently represents C 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, or a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0035] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, etc., and that is commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, which is incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by using other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, and 2-hydroxy-ethanesulfonate. , lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like.

[0036] Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, and N + (C 1-4Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Further pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed, where appropriate, using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates.

[0037] Unless otherwise stated, structures depicted herein are also meant to include all isomers (e.g., enantiomers, diastereomers, and geometric (or conformational) forms) of the structure, for example, R and S configurations for each asymmetric center, Z and E double bond isomers, and Z and E conformational isomers. Accordingly, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention. Unless otherwise stated, all tautomers of the present compounds are within the scope of the invention. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, replacement of hydrogen by deuterium or tritium, or 13 C or 14 Compounds having this structure including the replacement of a carbon with a C-enriched carbon are within the scope of the present invention. Such compounds are useful, for example, as analytical tools, probes in biological assays, or as therapeutic agents according to the present invention. In certain embodiments, the warhead moiety R of provided compounds is 1 contains one or more deuterium atoms. In certain embodiments, ring B of provided compounds can be substituted with one or more deuterium atoms.

[0038] Drawn structures represent relative configuration unless labeled as absolute configuration. The present invention contemplates individual enantiomers and racemic mixtures.

[0039] As used herein, an "HPK1 antagonist" or "HPK1 inhibitor" is a molecule that reduces, inhibits, or otherwise decreases one or more of HPK1's biological activities (e.g., serine / threonine kinase activity, recruitment to the TCR complex upon TCR activation, interaction with protein binding partners such as SLP76). Antagonism using an HPK1 antagonist does not necessarily indicate complete elimination of HPK1 activity. Instead, activity may be reduced by a statistically significant amount, including, for example, at least about a 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 95%, or 100% reduction in HPK1 activity compared to an appropriate control. In some embodiments, the HPK1 antagonist reduces, inhibits, or otherwise decreases the serine / threonine kinase activity of HPK1. In some of these embodiments, the HPK1 antagonist reduces, inhibits, or otherwise decreases HPK1-mediated phosphorylation of SLP76 and / or Gads. Compounds of the present disclosure directly bind to HPK1 and inhibit its kinase activity.

[0040] By "specific antagonist" is intended an agent that reduces, inhibits, or otherwise decreases the activity of a defined target to a greater extent than the activity of an unrelated target. For example, an HPK1-specific antagonist reduces at least one biological activity of HPK1 by an amount that is statistically greater than the inhibitory effect of the antagonist on any other protein (e.g., another serine / threonine kinase). In some embodiments, the IC of the antagonist against the target 50 is the IC of the antagonist against the non-target 50The IC50 of the HPK1 antagonist is about 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, 5%, 1%, 0.1%, 0.01%, 0.001%, or less. Compounds of the present disclosure may or may not be specific HPK1 antagonists. A specific HPK1 antagonist reduces the biological activity of HPK1 by an amount that is statistically greater than the inhibitory effect of the antagonist on any other protein (e.g., other serine / threonine kinase). In certain embodiments, the HPK1 antagonist specifically inhibits the serine / threonine kinase activity of HPK1. In some of these embodiments, the IC50 of the HPK1 antagonist for HPK1 is 50 The IC50 of HPK1 antagonists against other serine / threonine kinases or other types of kinases (e.g., tyrosine kinases) 50 is about 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, 0.1%, 0.01%, 0.001%, or less.

[0041] The compounds of the present invention may be linked to a detectable moiety. It will be understood that such compounds are useful as imaging agents. Those skilled in the art will recognize that the detectable moiety can be attached to a provided compound via a suitable substituent. As used herein, the term "suitable substituent" refers to a moiety capable of covalently binding to a detectable moiety. Such moieties are well known to those skilled in the art and include, for example, groups containing a carboxylate moiety, an amino moiety, a thiol moiety, or a hydroxyl moiety, to name a few. It will be understood that such moieties can be attached to a provided compound directly or via a linking group such as a divalent saturated or unsaturated hydrocarbon chain. In some embodiments, such moieties may be attached via click chemistry. In some embodiments, such moieties may be attached via 1,3-cycloaddition of an azide with an alkyne, optionally in the presence of a copper catalyst. Methods using click chemistry are known in the art and include those described by Rostovtsev et al., Angew. Chem. Int. Ed. 2002, 41, 2596-99, and Sun et al., Bioconjugate Chem., 2006, 17, 52-57.

[0042] As used herein, the term "detectable moiety" is used interchangeably with the term "label" and refers to any moiety that is capable of being detected, such as primary labels and secondary labels. Radioisotopes (e.g., tritium, 32 P, 33 P, 35 S, or 14 Primary labels, mass tags, and fluorescent labels such as C) are signal-generating reporter groups that can be detected without further modification. Detectable moieties also include luminescent and fluorescent groups.

[0043] As used herein, the term "secondary label" refers to moieties such as biotin and various protein antigens that require the presence of a second intermediate for the generation of a detectable signal. For biotin, the secondary intermediate may include a streptavidin-enzyme conjugate. For antigen labels, the secondary intermediate may include an antibody-enzyme conjugate. Some fluorescent groups act as secondary labels because they transfer energy to another group in the process of non-radiative fluorescence resonance energy transfer (FRET), and the second group generates the detected signal.

[0044] As used herein, the terms "fluorescent label," "fluorescent dye," and "fluorophore" refer to a moiety that absorbs light energy at a defined excitation wavelength and emits light energy at a different wavelength. Examples of fluorescent labels include, but are not limited to, Alexa Fluor dyes (Alexa Fluor 350, Alexa Fluor 488, Alexa Fluor 532, Alexa Fluor 546, Alexa Fluor 568, Alexa Fluor 594, Alexa Fluor 633, Alexa Fluor 660, and Alexa Fluor 680), AMCA, AMCA-S, BODIPY dyes (BODIPY FL, BODIPY R6G, BODIPY TMR, BODIPY TR, BODIPY 530 / 550, BODIPY 558 / 568, BODIPY 564 / 570, BODIPY 576 / 589, BODIPY 581 / 591, BODIPY 630 / 650, BODIPY 650 / 665), carboxyrhodamine 6G, carboxy-X-rhodamine (ROX), cascade blue, cascade yellow, coumarin 343, cyanine dyes (Cy3, Cy5, Cy3.5, Cy5.5), dansyl, dapoxyl, dialkylaminocoumarin, 4',5'-dichloro-2',7'-dimethoxy-fluorescein, DM-NERF, eosin, erythrosine, fluorescein, FAM, hydroxycoumarin, IRDyes (IRD40, IRD700, IRD800), JOE, Lisa Rhodamine B, Marine Blue, Methoxycoumarin, Naphthofluorescein, Oregon Green 488, Oregon Green 500, Oregon Green 514, Pacific Blue, PyMPO, Pyrene, Rhodamine B, Rhodamine 6G, Rhodamine Green, Rhodamine Red, Rhodol Green, 2',4',5',7'-tetra-bromosulfone-fluorescein, Tetramethyl-rhodamine (TMR), Carboxytetramethylrhodamine (TAMRA), Texas Red, Texas Red-X.

[0045] As used herein, the term "mass tag" refers to any moiety that can be uniquely detected by its mass using mass spectrometry (MS) detection techniques. Examples of mass tags include electrophoretic release tags such as N-[3-[4'-[(p-methoxytetrafluorobenzyl)oxy]phenyl]-3-methylglyceronyl]isonipecotic acid, 4'-[2,3,5,6-tetrafluoro-4-(pentafluorophenoxyl)]methylacetophenone, and derivatives thereof. The synthesis and utility of these mass tags are described in U.S. Patent Nos. 4,650,750, 4,709,016, 5,360,8191, 5,516,931, 5,602,273, 5,604,104, 5,610,020, and 5,650,270. Other examples of mass tags include, but are not limited to, nucleotides, dideoxynucleotides, oligonucleotides of various lengths and base compositions, oligopeptides, oligosaccharides, and other synthetic polymers of various lengths and monomer compositions. A wide variety of organic molecules, both neutral and charged (biomolecules or synthetic compounds) in the appropriate mass range (100-2000 daltons), can also be used as mass tags.

[0046] As used herein, the terms "measurable affinity" and "measurably inhibit" refer to a measurable change in HPK1 protein kinase activity between a sample containing a compound of the present invention, or a composition thereof, and HPK1 protein kinase, and an equivalent sample containing HPK1 protein kinase in the absence of the compound or composition thereof.

[0047] 3. Description of Exemplary Embodiments: As described above, in certain embodiments, the present invention provides compounds of formula I [ka] or a pharmaceutically acceptable salt thereof, wherein: Z is CR or N; X is a covalent bond, -O-, -S-, -NR-, -S(O)2-, -S(O)2NR-, -S(O)-, -S(O)NR-, -C(O)-, -C(O)O-, -C(O)NR-, -C(O)N(R)O-, -OC(O)-, -OC(O)NR-, -N(R)C(O)O-, -N(R)C(O)-, -N(R)S(O)2-, or X is C 1-4 a divalent saturated or unsaturated, straight or branched hydrocarbon chain in which one or two methylene units of the chain are optionally and independently replaced by -C(R)2-, -N(R)-, -N(R)C(O)-, -C(O)N(R)-, -N(R)S(O)2-, -S(O)2N(R)-, -O-, -C(O)-, -OC(O)-, -C(O)O-, -S-, -S(O)-, or -S(O)2-; R 1 But C 1-6 aliphatic; phenyl; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocyclic ring; a 5- to 6-membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 3- to 7-membered saturated or partially unsaturated monocyclic heterocyclic ring having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and an 8- to 10-membered saturated or partially unsaturated bicyclic heterocyclic ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from q R C has been replaced by the example R 2 But C 1-6 aliphatic; phenyl; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocyclic ring; a 5- to 6-membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and a 3- to 7-membered saturated or partially unsaturated monocyclic heterocyclic ring having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from q R C or R 2 -OR, -SR, -NR2, -S(O)2R, -S(O)2NR2, -S(O)R, -S(O)NR2, -C(O)R, -C(O)OR, -C(O)NR2, -C(O)N(R)OR, -OC(O)R, -OC(O)NR2, -N(R)C(O)OR, -N(R)C(O)R, -N(R)C(O)NR2, -N(R)C(NR)NR2, selected from -N(R)NR, -N(R)S(O)NR, -N(R)S(O)R, -N=S(O)R, -S(NR)(O)R, and -N(R)S(O)R; R 3 Each instance of is independently hydrogen or an optionally substituted C 1-6 is an aliphatic group, R C each instance of is independently oxo, halogen, -CN, -NO2, -OR, -SR, -NR2, -S(O)2R, -S(O)2NR2, -S(O)R, -S(O)NR2, -C(O)R, -C(O)OR, -C(O)NR2, -C(O)N(R)OR, -OC(O)R, -OC(O)NR2, -N(R)C(O)OR, -N(R)C(O)R, -N(R)C(O)NR2, -N(R)C(NR)NR2, -N(R)NR2, -N(R)S(O)2NR2, -N(R)S(O)2R, -N=S(O)R2, -S(NR)(O)R, -N(R)S(O)R, -N(R)CN, -P(O)(R)NR2, -P(O)(R)OR, or -P(O)R2, or R C Each instance of C 1-6aliphatic; phenyl; naphthalenyl; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocyclic ring; a 3- to 7-membered saturated or partially unsaturated monocyclic heterocyclic ring having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, phosphorus, silicon, and sulfur; or a 5- to 6-membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; an 8- to 10-membered bicyclic heteroaryl ring having 1 to 5 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 5- to 8-membered saturated or partially unsaturated bridged bicyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 6- to 11-membered saturated or partially unsaturated spirocyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or a 6- to 11-membered saturated or partially unsaturated bicyclic heterocyclic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from r instances of R and s instances of R. D has been replaced by the example R D each instance of is independently oxo, halogen, -CN, -NO2, -OR, -SR-, NR2, -S(O)2R, -S(O)2NR2, -S(O)R, -S(O)NR2, -C(O)R, -C(O)OR, -C(O)NR2, -C(O)N(R)OR, -OC(O)R, -OC(O)NR2, -N(R)C(O)OR, -N(R)C(O)R, -N(R)C(O)NR2, -N(R)C(NR)NR2, -N(R)NR2, -N(R)S(O)2NR2, -N(R)S(O)2R, -N=S(O)R2, -S(NR)(O)R, -N(R)S(O)R, -N(R)CN, -P(O)(R)NR2, -P(O)(R)OR, or -P(O)R2; Each R is independently hydrogen, —CN, halogen, or C 1-6aliphatic; phenyl; naphthalenyl; 3- to 7-membered saturated or partially unsaturated monocyclic carbocyclic ring; 3- to 7-membered saturated or partially unsaturated monocyclic heterocyclic ring having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur; 5- to 6-membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; 8- to 10-membered bicyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; an optionally substituted group selected from: a 7- to 12-membered saturated or partially unsaturated bicyclic heterocycle having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 5- to 8-membered saturated or partially unsaturated bridged bicyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 6- to 10-membered saturated or partially unsaturated spirocycle having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or a 6- to 11-membered saturated or partially unsaturated bicyclic carbocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur; two R groups on the same nitrogen, taken together with the nitrogen, form an optionally substituted 4-7 membered monocyclic saturated, partially unsaturated, or heteroaryl ring having, in addition to the nitrogen, 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; an 8-10 membered bicyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or a 7-12 membered saturated or partially unsaturated bicyclic heterocycle having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; m is 0, 1, or 2; each q is independently 0, 1, 2, 3, or 4; each r is independently 0, 1, 2, 3, or 4; Compounds of Formula I, or pharmaceutically acceptable salts thereof, are provided wherein each s is independently 0, 1, 2, 3, or 4.

[0048] Z is CR or N, as generally defined above.

[0049] In some embodiments, Z is CR. In some embodiments, Z is N. In some embodiments, Z is CH.

[0050] In some embodiments, Z is selected from those shown in Table 1 below.

[0051] As generally defined above, X is a covalent bond, —O—, —S—, —NR—, —S(O)—, —S(O)NR—, —S(O)—, —S(O)NR—, —C(O)—, —C(O)O—, —C(O)NR—, —C(O)N(R)O—, —OC(O)—, —OC(O)NR—, —N(R)C(O)O—, —N(R)C(O)—, —N(R)S(O)—, or X is C 1-4 A divalent saturated or unsaturated, straight or branched hydrocarbon chain in which one or two methylene units of the chain are optionally and independently replaced by -C(R)2-, -N(R)-, -N(R)C(O)-, -C(O)N(R)-, -N(R)S(O)2-, -S(O)2N(R)-, -O-, -C(O)-, -OC(O)-, -C(O)O-, -S-, -S(O)-, or -S(O)2-.

[0052] In certain embodiments, X is —O—, —S—, —NR—, —S(O)—, —S(O)NR—, —S(O)—, —S(O)NR—, —C(O)—, —C(O)O—, —C(O)NR—, —C(O)N(R)O—, —OC(O)—, —OC(O)NR—, —N(R)C(O)O—, —N(R)C(O)—, —N(R)C(O)NR—, —N(R)C(NR)NR—, —N(R)NR—, —N(R)S(O)NR—, or —N(R)S(O)—.

[0053] In some embodiments, X is C 1-4A divalent saturated or unsaturated, straight or branched hydrocarbon chain in which one or two methylene units of the chain are optionally and independently replaced by -C(R)2-, -N(R)-, -N(R)C(O)-, -C(O)N(R)-, -N(R)S(O)2-, -S(O)2N(R)-, -O-, -C(O)-, -OC(O)-, -C(O)O-, -S-, -S(O)-, or -S(O)2-.

[0054] In certain embodiments, X is -NR-, -C(O)-, -C(O)O-, -C(O)NR-, -C(O)N(R)O-, -OC(O)-, -OC(O)NR-, -N(R)C(O)O-, -N(R)C(O)-, -N(R)C(O)NR-, -N(R)C(NR)NR-, or -N(R)NR-.

[0055] In certain embodiments, X is -NR-. In certain embodiments, X is -NH-.

[0056] In some embodiments, X is selected from those shown in Table 1 below.

[0057] Generally, as defined above, R 1 is C 1-6 aliphatic; phenyl; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocyclic ring; a 5- to 6-membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 3- to 7-membered saturated or partially unsaturated monocyclic heterocyclic ring having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and an 8- to 10-membered saturated or partially unsaturated bicyclic heterocyclic ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from q R C has been replaced with the example.

[0058] In some embodiments, R 1 But there are q R C Example of C being substituted 1-6 Aliphatic; q R C Examples of phenyl substituted with q R Cq R C a 5- to 6-membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, substituted with, for example, q R C a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, substituted with, for example, q R C and R is an 8- to 10-membered saturated or partially unsaturated bicyclic heterocycle having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, substituted with, for example,

[0059] In some embodiments, R 1 is phenyl or a 5- to 6-membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from q R C has been replaced with the example.

[0060] In certain embodiments, R 1is phenyl, indanyl, tetrahydronaphthyl, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, NH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydro Furan, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isoindolinyl, isoindolenyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl;-1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, oxetanyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl thiazolyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, azetidinyl, or xanthenyl, each of which is q R; C has been replaced with the example.

[0061] In certain embodiments, R 1is phenyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, isothiazolyl, isoxazolyl, morpholinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl; -1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, oxetanyl, pyrimidinyl, piperazinyl, piperidinyl, pyranyl, pyrazinyl, piperazine, pyrazinyl ... oxazolyl, oxazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, oxetanyl, pyrimidinyl, oxazolyl, oxazolyl, oxazolidinyl, oxetanyl, pyrimidinyl, piperazinyl, piperidinyl, pyrazinyl, pyrazinyl, pyrazinyl, pyrazinyl, pyrazinyl, pyrazinyl, pyrazinyl, pyrazinyl, oxazolyl, oxazolyl, oxazolidine, oxazolyl, oxazolidinyl, oxazolidine, oxazolyl, oxazolidinyl, oxetanyl, pyrimidinyl, piperazinyl, piperazine, pyrazinyl, pyrazinyl, pyrazinyl, pyrazinyl, thiazolyl, thiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, azetidinyl, or xanthenyl, each of which is selected from the group consisting of q R C has been replaced by the example

[0062] In certain embodiments, R 1 is furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl; -1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, oxetanyl, pyrimidinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, thiazolyl, thienyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, or 1,2,5-triazolyl, 1,3,4-triazolyl, each of which is q R C has been replaced by the example

[0063] In certain embodiments, R 1 is phenyl, pyrazolyl, pyridinyl, pyrazinyl, or pyrimidinyl, each of which is selected from the group consisting of q R C has been replaced by the example

[0064] In certain embodiments, R 1 is pyrazolyl or pyridinyl, each of which is q R C has been replaced by the example

[0065] In certain embodiments, R 1 is pyrazolyl or pyridinyl, each of which is q R C are replaced by examples, and each R C are independently halogen, —CN, —OR, —S(O)R, —C(O)NR, or R C Each instance of 1-6 an optionally substituted group selected from aliphatic; a 5-10 membered saturated or partially unsaturated bridged bicyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 6-12 membered saturated or partially unsaturated spirocyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 3-7 membered saturated or partially unsaturated monocyclic carbocyclic ring; a 3-7 membered saturated or partially unsaturated monocyclic heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, phosphorus, silicon, and sulfur, or two R C groups, taken together with the atoms to which they are each attached, form a bridged, fused, or spiro 5- to 6-membered aryl ring, a 3- to 7-membered saturated or partially unsaturated monocyclic carbocycle, or a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, phosphorus, silicon, and sulfur; R C Each instance of is independently R and R D was optionally replaced by

[0066] In certain embodiments, R 1 teeth, [ka] is.

[0067] In certain embodiments, R 1 teeth, [ka] is.

[0068] In certain embodiments, R 1 teeth, [ka] and In the formula, R C Each instance of is independently a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, phosphorus, silicon, and sulfur; or a 5- to 8-membered saturated or partially unsaturated bridged bicyclic ring having 0 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is independently selected from r instances of R and s instances of R. D has been replaced by the example

[0069] In certain embodiments, R 1 teeth, [ka] and In the formula, R C Each example is independently -Me, -Et, -CN, -F, -OMe, -S(O)2Me, [ka] [ka] is.

[0070] In certain embodiments, R 1 is that R C Together with the substituents, [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] is.

[0071] In certain embodiments, R 1 together with its R-C substituents, [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] is.

[0072] In certain embodiments, R 1 is that R C Together with the substituents, [ka] [ka] [ka] is.

[0073] In some embodiments, R 1 teeth, [ka] [ka] is.

[0074] In some embodiments, R 1 is selected from those shown in Table 1 below.

[0075] In some embodiments, R 2 is C 1-6aliphatic; phenyl; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocyclic ring; a 5- to 6-membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and a 3- to 7-membered saturated or partially unsaturated monocyclic heterocyclic ring having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from q R C or replaced by examples of R 2 is selected from —OR, —SR, —NR2, —S(O)2R, —S(O)2NR2, —S(O)R, —S(O)NR2, —C(O)R, —C(O)OR, and —C(O)NR2.

[0076] In certain embodiments, R 2 is methyl, ethyl, n-propyl, i-Pr, n-Bu, s-Bu, t-Bu, straight-chain or branched pentyl, straight-chain or branched hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, phenyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, isothiazolyl, isoxazolyl, morpholinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl; 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, oxetanyl, pyrimidinyl, piperazinyl, piperidinyl nyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridinyl, pyridyl, pyridin-one, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, tetrahydropyranyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4 thiadiazolyl, thienyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, or azetidinyl, each of which is selected from the group consisting of q R C or replaced by examples of R 2is selected from —OR, —SR, —NR2, —S(O)2R, —S(O)2NR2, —S(O)R, —S(O)NR2, —C(O)R, —C(O)OR, and —C(O)NR2.

[0077] In certain embodiments, R 2 is methyl, cyclopropyl, phenyl, imidazolyl, morpholinyl, oxazolyl, pyrazolyl, pyridazinyl, pyridinyl, pyridyl, pyridin-one, pyrimidinyl, pyrrolyl, or tetrahydropyranyl, each of which is selected from the group consisting of q R C or replaced by examples of R 2 is selected from —S(O)R and —C(O)NR.

[0078] In certain embodiments, R 2 teeth, [ka] [ka] is.

[0079] In certain embodiments, R 2 is that R C Together with the substituents, [ka] [ka] [ka] is.

[0080] In certain embodiments, R 2 is that R C Together with the substituents, [ka] [ka] [ka] [ka] , is.

[0081] In some embodiments, R 2 is selected from those shown in Table 1 below.

[0082] Generally, as defined above, R 3 Each instance of is independently hydrogen or an optionally substituted C 1-6 It is an aliphatic group.

[0083] In some embodiments, R 3 is hydrogen.

[0084] In some embodiments, R 3 is an arbitrarily substituted C 1-6 It is an aliphatic group.

[0085] In some embodiments, R 3 is methyl. In some embodiments, R 3 teeth, [ka] In some embodiments, R 3 teeth, [ka] is.

[0086] In some embodiments, R 3 is selected from those shown in Table 1 below.

[0087] Generally, as defined above, R C Each instance of is independently oxo, halogen, -CN, -NO2, -OR, -SR, -NR2, -S(O)2R, -S(O)2NR2, -S(O)R, -S(O)NR2, -C(O)R, -C(O)OR, -C(O)NR2, -C(O)N(R)OR, -OC(O)R, -OC(O)NR2, -N(R)C(O)OR, -N(R)C(O)R, -N(R)C(O)NR2, -N(R)C(NR)NR2, -N(R)NR2, -N(R)S(O)2NR2, -N(R)S(O)2R, -N=S(O)R2, -S(NR)(O)R, -N(R)S(O)R, -N(R)CN, -P(O)(R)NR2, -P(O)(R)OR, or -P(O)R2, or R C Each instance of C 1-6 aliphatic; phenyl; naphthalenyl; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocyclic ring; a 3- to 7-membered saturated or partially unsaturated monocyclic heterocyclic ring having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, phosphorus, silicon, and sulfur; or a 5- to 6-membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; an 8- to 10-membered bicyclic heteroaryl ring having 1 to 5 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and optionally substituted groups selected from: a 5-8 membered saturated or partially unsaturated bridged bicyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 6-10 membered saturated or partially unsaturated spiro ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or a 6-11 membered saturated or partially unsaturated bicyclic heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from r instances of R and s instances of R. D has been replaced with the example.

[0088] In some embodiments, R C Each instance of is independently oxo, halogen, -CN, -NO2, -OR, -SR, -NR2, -S(O)2R, -S(O)2NR2, -S(O)R, -S(O)NR2, -C(O)R, -C(O)OR, -C(O)NR2, -C(O)N(R)OR, -OC(O)R, -OC(O)NR2, -N(R)C(O)OR, -N(R)C(O)R, -N(R)C(O)NR2, -N(R)C(NR)NR2, -N(R)NR2, -N(R)S(O)2NR2, -N(R)S(O)2R, -N=S(O)R2, -S(NR)(O)R, -N(R)S(O)R, -N(R)CN, -P(O)(R)NR2, -P(O)(R)OR, or -P(O)R2, or R C Each instance of C 1-6 an optionally substituted group selected from aliphatic; phenyl; naphthalenyl; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocycle; a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, phosphorus, silicon, and sulfur; or a 5- to 6-membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; an 8- to 10-membered bicyclic heteroaryl ring having 1 to 5 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 5- to 8-membered saturated or partially unsaturated bridged bicyclic ring having 0 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 6- to 10-membered saturated or partially unsaturated spirocycle having 0 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or a 6- to 11-membered saturated or partially unsaturated bicyclic heterocycle having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0089] In some embodiments, R C Each example is independently oxo, halogen, -CN, -NO2, -OR, -SR-, NR2, -S(O)2R, -S(O)2NR2, -S(O)R, -S(O)NR2, -C(O)R, -C(O)OR, -C(O)NR2, -C(O)N(R)OR, -OC(O)R, -OC(O)NR2, -N(R)C(O)OR, -N(R)C(O)R, -N(R)C(O)NR2, -N(R)C(NR)NR2, -N(R)NR2, -N(R)S(O)2NR2, -N(R)S(O)2R, -N=S(O)R2, -S(NR)(O)R, -N(R)S(O)R, -N(R)CN, -P(O)(R)NR2, -P(O)(R)OR, or -P(O)R2.

[0090] In some embodiments, R C Each instance of 1-6 aliphatic; phenyl; naphthalenyl; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocyclic ring; a 3- to 7-membered saturated or partially unsaturated monocyclic heterocyclic ring having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, phosphorus, silicon, and sulfur; or a 5- to 6-membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; an 8- to 10-membered bicyclic heteroaryl ring having 1 to 5 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 5- to 8-membered saturated or partially unsaturated bridged bicyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 6- to 10-membered saturated or partially unsaturated spirocyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or a 6- to 11-membered saturated or unsaturated bicyclic heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0091] In some embodiments, each R C are 6- to 11-membered saturated or partially unsaturated spiro rings having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and are r instances of R and s instances of R D has been replaced with the example.

[0092] In some embodiments, each R C are 6- to 11-membered saturated or partially unsaturated bicyclic heterocycles having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and are r instances of R and s instances of R Dhas been replaced with the example.

[0093] In some embodiments, each R C is independently methyl, oxo, fluoro or methoxy.

[0094] In some embodiments, each R C is, independently, [ka] [ka] [ka] [ka] is.

[0095] In some embodiments, each R C is independently -CHF2 or chloro.

[0096] In some embodiments, each R C is, independently, [ka] [ka] [ka] [ka] [ka] [ka] is.

[0097] In some embodiments, each R C is, independently, [ka] [ka] is.

[0098] In certain embodiments, each R C are independently -Me, -Et, -F, -OMe, -NH2, -NHMe, -N(Me)2, [ka] is.

[0099] In some embodiments, R C Each example is selected from those shown in Table 1 below.

[0100] Generally, as defined above, R D Each example is independently oxo, halogen, -CN, -NO2, -OR, -SR-, NR2, -S(O)2R, -S(O)2NR2, -S(O)R, -S(O)NR2, -C(O)R, -C(O)OR, -C(O)NR2, -C(O)N(R)OR, -OC(O)R, -OC(O)NR2, -N(R)C(O)OR, -N(R)C(O)R, -N(R)C(O)NR2, -N(R)C(NR)NR2, -N(R)NR2, -N(R)S(O)2NR2, -N(R)S(O)2R, -N=S(O)R2, -S(NR)(O)R, -N(R)S(O)R, -N(R)CN, -P(O)(R)NR2, -P(O)(R)OR, or -P(O)R2.

[0101] In some embodiments, R D is oxo, halogen, -CN, -NO2, -OR, -SR-, NR2, -S(O)2R, -S(O)2NR2, -S(O)R, -S(O)NR2, -C(O)R, -C(O)OR, -C(O)NR2, -C(O)N(R)OR, -OC(O)R, -OC(O)NR2, -N(R)C(O)OR, -N(R)C(O)R, -N(R)C(O)NR2, -N(R)C(NR)NR2, -N(R)NR2, -N(R)S(O)2NR2, -N(R)S(O)2R, -N=S(O)R2, -S(NR)(O)R, -N(R)S(O)R, -N(R)CN, -P(O)(R)NR2, -P(O)(R)OR, or -P(O)R2.

[0102] In some embodiments, R D is hydroxy, fluoro, or methoxy.

[0103] In some embodiments, R D teeth, [ka] is.

[0104] In some embodiments, R D is oxo.

[0105] In some embodiments, R D teeth, [ka] is.

[0106] In some embodiments, R D is selected from those shown in Table 1 below.

[0107] As generally defined above, each R is independently hydrogen or C 1-6aliphatic; phenyl; naphthalenyl; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocycle; a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 5- to 6-membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; an 8- to 10-membered bicyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 7- to 12-membered saturated or partially unsaturated bicyclic heterocycle having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 5- to 8-membered saturated or partially unsaturated bridged bicyclic ring having 0 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; an optionally substituted group selected from a 6- to 10-membered saturated or partially unsaturated spirocycle having 0-3 heteroatoms; a 6- to 11-membered saturated or partially unsaturated bicyclic carbocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or two R groups on the same nitrogen, taken together with the nitrogen, form an optionally substituted 4- to 7-membered monocyclic saturated, partially unsaturated, or heteroaryl ring having, in addition to the nitrogen, 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; an 8- to 10-membered bicyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or a 7- to 12-membered saturated or partially unsaturated bicyclic heterocycle having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0108] In some embodiments, R is methyl. In some embodiments, R is [ka] is.

[0109] In some embodiments, R is ethyl.

[0110] In some embodiments, R is [ka] [ka] is.

[0111] In some embodiments, R is selected from those shown in Table 1 below.

[0112] As generally defined above, each hydrogen bonded to a carbon can be optionally and independently replaced by deuterium.

[0113] In some embodiments, a hydrogen bonded to a carbon is replaced by deuterium.

[0114] Generally, as defined above, m is 0, 1, or 2.

[0115] In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2.

[0116] In some embodiments, m is selected from those shown in Table 1 below.

[0117] As generally defined above, q is 0, 1, 2, 3, or 4. In some embodiments, q is 0. In some embodiments, q is 1, 2, 3, or 4. In some embodiments, q is 1. In some embodiments, q is 2. In some embodiments, q is 3. In some embodiments, q is 4.

[0118] In some embodiments, q is 1, 2, or 3. In some embodiments, q is 1 or 2.

[0119] In some embodiments, q is selected from those shown in Table 1 below.

[0120] As generally defined above, r is 0, 1, 2, 3, or 4. In some embodiments, r is 0. In some embodiments, r is 1, 2, 3, or 4. In some embodiments, r is 1. In some embodiments, r is 2. In some embodiments, r is 3. In some embodiments, r is 4.

[0121] In some embodiments, r is 1 or 2. In some embodiments, r is 2 or 3. In some embodiments, r is 2, 3, or 4.

[0122] In some embodiments, r is selected from those shown in Table 1 below.

[0123] As generally defined above, s is 0, 1, 2, 3, or 4. In some embodiments, s is 0. In some embodiments, s is 1, 2, 3, or 4. In some embodiments, s is 1. In some embodiments, s is 2. In some embodiments, s is 3. In some embodiments, s is 4.

[0124] In some embodiments, s is 1 or 2. In some embodiments, s is 2 or 3. In some embodiments, s is 2, 3, or 4.

[0125] In some embodiments, s is selected from those shown in Table 1 below.

[0126] In some embodiments, the present invention provides a compound of formula II [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 and R 2 Each of the following, both alone and in combination, is as defined above and described in the embodiments herein.

[0127] In some embodiments, the present invention provides a compound of formula III [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 and R 2 Each of the following, both alone and in combination, is as defined above and described in the embodiments herein.

[0128] In some embodiments, the present invention provides a compound of formula IV [ka] or a pharmaceutically acceptable salt thereof, wherein R 2 and R C Each of the following, both alone and in combination, is as defined above and described in the embodiments herein.

[0129] In some embodiments, the present invention provides a compound of formula V [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 and X, both alone and in combination, are as defined above and described in the embodiments herein.

[0130] In some embodiments, the present invention provides a compound of formula VI [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 and R 2 Each of the following, both alone and in combination, is as defined above and described in the embodiments herein.

[0131] In some embodiments, the present invention provides a compound of formula VII [ka] or a pharmaceutically acceptable salt thereof, wherein R 2 and R C Each of the following, both alone and in combination, is as defined above and described in the embodiments herein.

[0132] Exemplary compounds of the present invention are shown in Table 1 below. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11] [Table 1-12] [Table 1-13]

[0133] In some embodiments, the present invention provides a compound shown in Table 1 above, or a pharmaceutically acceptable salt thereof. In some embodiments, the present invention provides a compound shown in Table 1 above. In some embodiments, the present invention provides a pharmaceutical composition comprising a compound shown in Table 1 above, or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier, excipient, or diluent.

[0134] In some embodiments, the present invention provides a compound of formula I as defined above, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula I as defined above, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle, for use as a medicament.

[0135] In some embodiments, the present invention also provides a compound of formula I as described herein or a pharmaceutical composition as described herein for use in a method of inhibiting HPK1 as described herein, in a method for enhancing an immune response as described herein in a subject in need thereof, and / or in a method for treating an HPK1-dependent disorder as described herein.

[0136] In some embodiments, the present invention also provides a compound of formula I as described herein or a pharmaceutical composition as described herein for use in a method for inhibiting HPK1, as described herein.

[0137] In some embodiments, the present invention also provides a compound of formula I as described herein or a pharmaceutical composition as described herein for use in a method for enhancing an immune response in a subject in need thereof, as described herein.

[0138] In some embodiments, the present invention also provides a compound of formula I as described herein or a pharmaceutical composition as described herein for use in a method for treating an HPK1-dependent disorder, as described herein.

[0139] In some embodiments, the present invention also provides the use of a compound of formula I as described herein or a pharmaceutical composition as described herein for the manufacture of a medicament for inhibiting HPK1, enhancing an immune response in a subject in need thereof, and / or treating an HPK1-dependent disorder.

[0140] In some embodiments, the present invention also provides the use of a compound of Formula I described herein or a pharmaceutical composition described herein for the manufacture of a medicament for inhibiting HPK1.

[0141] In some embodiments, the present invention also provides the use of a compound of formula I as described herein or a pharmaceutical composition as described herein for the manufacture of a medicament for enhancing an immune response in a subject in need thereof.

[0142] In some embodiments, the present invention also provides the use of a compound of Formula I described herein or a pharmaceutical composition described herein for the manufacture of a medicament for treating an HPK1-dependent disorder.

[0143] In some embodiments, the present invention also provides for the use of a compound of formula I as described herein or a pharmaceutical composition as described herein in a method for inhibiting HPK1 as described herein, in a method for enhancing an immune response as described herein in a subject in need thereof, and / or in a method for treating an HPK1-dependent disorder as described herein.

[0144] In some embodiments, the present invention also provides the use of a compound of Formula I as described herein or a pharmaceutical composition as described herein in a method for inhibiting HPK1, as described herein.

[0145] In some embodiments, the present invention also provides the use of a compound of formula I as described herein or a pharmaceutical composition as described herein in a method for enhancing an immune response in a subject in need thereof.

[0146] In some embodiments, the present invention also provides the use of a compound of formula I as described herein or a pharmaceutical composition as described herein in a method for treating an HPK1-dependent disorder, as described herein.

[0147] 4. General Methods of Providing Compounds of the Invention The compounds of the invention may generally be prepared or isolated by synthetic and / or semi-synthetic methods known to those skilled in the art for similar compounds, and by methods detailed in the Examples herein.

[0148] 5. Use, Formulation, and Administration Pharmaceutically acceptable compositions According to another embodiment, the present invention provides a composition comprising a compound of the present invention or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant, or vehicle. The amount of compound in the composition of the present invention is an amount effective to measurably inhibit HPK1 or a mutant thereof in a biological sample or in a patient. In certain embodiments, the amount of compound in the composition of the present invention is an amount effective to measurably inhibit HPK1 or a mutant thereof in a biological sample or in a patient. In certain embodiments, the composition of the present invention is formulated for administration to a patient in need of such a composition. In some embodiments, the composition of the present invention is formulated for oral administration to a patient.

[0149] The term "patient", as used herein, means an animal, preferably a mammal, and most preferably a human.

[0150] The term "pharmaceutically acceptable carrier, adjuvant, or vehicle" refers to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound in which it is formulated. Pharmaceutically acceptable carriers, adjuvants, or vehicles that can be used in the compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and wool fat.

[0151] "Pharmaceutically acceptable derivative" means any non-toxic salt, ester, salt of an ester, or other derivative of a compound of this invention that is capable of providing, either directly or indirectly, a compound of this invention or an inhibitory active metabolite or residue thereof, upon administration to a recipient.

[0152] As used herein, the term "inhibitorily active metabolite or residue thereof" means that a metabolite or residue thereof is also an inhibitor of HPK1 or a mutant thereof.

[0153] The subject matter disclosed herein includes prodrugs, metabolites, derivatives, and pharmaceutically acceptable salts of the compounds of the present invention. Metabolites include compounds produced by a process comprising contacting a compound of the present invention with a mammal for a period of time sufficient to produce its metabolic product. When the compound of the present invention is a base, the desired pharmaceutically acceptable salt can be prepared by any suitable method available in the art, for example, by treating the free acid with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methanesulfonic acid, phosphoric acid, or an organic acid such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid such as glucuronic acid or galacturonic acid, an alpha hydroxy acid such as citric acid or tartaric acid, an amino acid such as aspartic acid or glutamic acid, an aromatic acid such as benzoic acid or cinnamic acid, or a sulfonic acid such as p-toluenesulfonic acid or ethanesulfonic acid. If the compound of the present invention is an acid, the desired pharmaceutically acceptable salt can be prepared by any suitable method, for example, treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary, tertiary), alkali metal hydroxide, or alkaline earth metal hydroxide. Illustrative examples of suitable salts include, but are not limited to, organic salts derived from amino acids such as glycine and arginine, ammonia, primary amines, secondary amines, and tertiary amines, and cyclic amines such as piperidine, morpholine, and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum, and lithium.

[0154] The compounds of the present invention may be in the form of a "prodrug," including compounds having a moiety that can be metabolized in vivo. Generally, prodrugs are metabolized in vivo by esterases or other mechanisms to active drugs. Examples of prodrugs and their uses are well known in the art (see, for example, Berge et al. (1977) "Pharmaceutical Salts," J. Pharm. Sci. 66:1-19). Prodrugs can be prepared in situ during the final isolation and purification of a compound, or by separately reacting a purified compound in its free acid form or a hydroxyl with a suitable esterifying agent. A hydroxyl group can be converted to an ester via treatment with a carboxylic acid. Examples of prodrug moieties include substituted and unsubstituted, branched or unbranched lower alkyl ester moieties (e.g., propionate esters), lower alkenyl esters, di-lower alkyl-amino lower alkyl esters (e.g., dimethylaminoethyl esters), acylamino lower alkyl esters (e.g., acetyloxymethyl esters), acyloxy lower alkyl esters (e.g., pivaloyloxymethyl esters), aryl esters (phenyl esters), aryl-lower alkyl esters (e.g., benzyl esters), substituted (e.g., with methyl, halo, or methoxy substituents) aryl and aryl-lower alkyl esters, amides, lower alkyl amides, di-lower alkyl amides, and hydroxyamides. Prodrugs that are converted to active forms in vivo through other mechanisms are also included. In an embodiment, the compounds of the present invention are prodrugs of any of the formulas herein.

[0155] The compositions of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or via an implanted reservoir. As used herein, the term "oral" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally, or intravenously. Sterile injectable forms of the compositions of the present invention may be aqueous or oily suspensions. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations may also be sterile injectable solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, for example, as solutions in 1,3-butanediol. Among the acceptable vehicles and solvents that may be used are water, Ringer's solution, and isotonic sodium chloride solution. Additionally, sterile, fixed oils are conventionally used as solvents or suspending media.

[0156] For this purpose, any bland fixed oil can be used, including synthetic monoglycerides or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives, especially in their polyoxyethylated versions, are useful for preparing injections, as are natural pharmaceutically acceptable oils such as olive oil or castor oil. These oil solutions or suspensions can also contain long-chain alcohol diluents or dispersants such as carboxymethylcellulose, or similar dispersants commonly used in the formulation of pharmaceutically acceptable dosage forms, including emulsions and suspensions. Other commonly used surfactants, such as Tweens, Spans, and other emulsifiers or bioavailability enhancers, commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms, can also be used for formulation purposes.

[0157] The pharmaceutically acceptable composition of the present invention can be orally administered in any orally acceptable dosage form, including but not limited to capsules, tablets, aqueous suspensions or solutions.For tablets for oral use, commonly used carriers include lactose and corn starch.Lubricants such as magnesium stearate are also typically added.For oral administration in capsule form, useful diluents include lactose and dried corn starch.When aqueous suspensions are required for oral use, active ingredients are combined with emulsifiers and suspending agents.If desired, certain sweeteners, flavorings or coloring agents can also be added.

[0158] Alternatively, the pharmaceutically acceptable compositions of the present invention may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature, and therefore melts in the rectum to release the drug. Such materials include cocoa butter, beeswax, and polyethylene glycol.

[0159] The pharmaceutically acceptable compositions of this invention may also be administered topically, particularly when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower gastrointestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.

[0160] Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches may also be used.

[0161] For topical application, the provided pharmaceutically acceptable compositions can be formulated into a suitable ointment containing the active ingredient suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of the present invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsifying wax, and water. Alternatively, the provided pharmaceutically acceptable compositions can be formulated into a suitable lotion or cream containing the active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and water.

[0162] For ophthalmic use, the provided pharmaceutically acceptable compositions can be formulated as a micronized suspension in isotonic, pH-adjusted, sterile saline, or preferably as a solution in isotonic, pH-adjusted, sterile saline, either with or without a preservative, such as benzylalkonium chloride. Alternatively, for ophthalmic use, the pharmaceutically acceptable compositions can be formulated into an ointment, such as petrolatum.

[0163] The pharmaceutically acceptable compositions of this invention may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation, and may be prepared as solutions in saline using benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents.

[0164] Most preferably, the pharmaceutically acceptable compositions of the present invention are formulated for oral administration. Such formulations can be administered with or without food. In some embodiments, the pharmaceutically acceptable compositions of the present invention are administered without food. In other embodiments, the pharmaceutically acceptable compositions of the present invention are administered with food.

[0165] The amount of the compounds of the present invention that may be combined with the carrier materials to produce a composition in a single dosage form will vary depending on the host being treated and the particular mode of administration. Preferably, the provided compositions should be formulated so that a dosage of 0.01 to 100 mg / kg body weight / day of the inhibitor can be administered to a patient receiving these compositions.

[0166] It should also be understood that the specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, excretion rate, drug combination, as well as the judgment of the treating physician and the severity of the particular disease being treated. The amount of a compound of the invention in a composition will also depend on the particular compound in the composition.

[0167] Uses of the Compounds and Pharmaceutically Acceptable Compositions The compounds and compositions described herein are generally useful for inhibiting the kinase activity of one or more enzymes. In some embodiments, the kinase inhibited by the compounds and methods of the invention is HPK1.

[0168] The compounds of the present disclosure find use in inhibiting the activity of the enzyme HPK1. HPK1 is a member of the germinal center kinase subfamily of Ste20-related serine / threonine kinases. HPK1 functions as a MAP4K by phosphorylating and activating MAP3K proteins, including MEKK1, MLK3, and TAK1, leading to activation of the MAPK Jnk.

[0169] In one embodiment, the presently disclosed subject matter is directed to a method of inhibiting HPK1, the method comprising contacting HPK1 with an effective amount of a compound of the invention or a pharmaceutical composition described herein.

[0170] In certain embodiments, the presently disclosed subject matter is directed to a method for enhancing an immune response in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of the invention or a pharmaceutical composition described herein. In certain aspects of this embodiment, T cells in the subject have at least one of enhanced priming, enhanced activation, enhanced migration, enhanced proliferation, enhanced survival, and enhanced cytolytic activity relative to before administration of the compound or pharmaceutical composition. In certain aspects of this embodiment, T cell activation is characterized by an elevated frequency of γ-IFN+ CD8 T cells or enhanced levels of IL-2 or granzyme B production by the T cells relative to before administration of the compound or pharmaceutical composition. In certain aspects of this embodiment, the number of T cells is increased relative to before administration of the compound or pharmaceutical composition. In certain aspects of this embodiment, the T cells are antigen-specific CD8 T cells. In certain aspects of this embodiment, antigen-presenting cells in the subject have enhanced maturation and activation relative to before administration of the compound or pharmaceutical composition. In certain aspects of this embodiment, the antigen-presenting cells are dendritic cells. In certain aspects of this embodiment, antigen-presenting cell maturation is characterized by an increased frequency of CD83+ dendritic cells. In certain aspects of this embodiment, antigen-presenting cell activation is characterized by elevated expression of CD80 and CD86 on dendritic cells.

[0171] Compounds of the present disclosure directly bind to and inhibit the kinase activity of HPK1, hi some embodiments, compounds of the present disclosure reduce, inhibit, or otherwise decrease HPK1-mediated phosphorylation of SLP76 and / or Gads.

[0172] The compounds of the present disclosure may or may not be specific HPK1 antagonists. A specific HPK1 antagonist reduces the biological activity of HPK1 by an amount that is statistically greater than the inhibitory effect of the antagonist on any other protein (e.g., other serine / threonine kinase). In certain embodiments, the compounds of the present disclosure specifically inhibit the serine / threonine kinase activity of HPK1. In some of these embodiments, the IC of the HPK1 antagonist for HPK1 is 50 The IC50 of HPK1 antagonists against other serine / threonine kinases or other types of kinases (e.g., tyrosine kinases) 50 is about 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, 0.1%, 0.01%, 0.001%, or less.

[0173] The compounds of the present disclosure can be used in methods for inhibiting HPK1. Such methods include contacting HPK1 with an effective amount of a compound of the present disclosure. By "contacting" is intended bringing the compound within sufficient proximity of an isolated HPK1 enzyme or a cell expressing HPK1 (e.g., a T cell, a B cell, a dendritic cell) so that the compound can bind to and inhibit the activity of HPK1. The compound can be contacted with HPK1 in vitro or in vivo via administration of the compound to a subject.

[0174] Any method known in the art for measuring the kinase activity of HPK1 may be used to determine whether HPK1 is inhibited, including in vitro kinase assays, immunoblots with antibodies specific for phosphorylated targets of HPK1 such as SLP76 and Gads, or measurement of downstream biological effects of HPK1 kinase activity, such as recruitment of 14-3-3 proteins to phosphorylated SLP7 and Gads, release of SLP76-Gads-14-3-3 complexes from LAT-containing microclusters, or activation of T cells or B cells.

[0175] The compounds of the present disclosure can be used to treat HPK1-dependent disorders. As used herein, an "HPK1-dependent disorder" is a pathological condition in which HPK1 activity is required for the development or maintenance of the pathological condition. In some embodiments, the HPK1-dependent disorder is cancer.

[0176] The compounds of the present disclosure also find use in enhancing an immune response in a subject in need thereof. Such a method comprises administering an effective amount of a compound of the present invention.

[0177] As used herein, "enhancing an immune response" refers to any improvement in an immunogenic response to an antigen. Non-limiting examples of improved immunogenic responses to an antigen include enhanced maturation or migration of dendritic cells, enhanced activation of T cells (e.g., CD4 T cells, CD8 T cells), enhanced T cell (e.g., CD4 T cells, CD8 T cell) proliferation, enhanced B cell proliferation, increased survival of T cells and / or B cells, improved antigen presentation by antigen-presenting cells (e.g., dendritic cells), improved antigen clearance, increased cytokine production by T cells (e.g., interleukin-2), increased resistance to prostaglandin E2-induced immunosuppression, and / or increased priming and / or cytolytic activity of CD8 T cells.

[0178] In some embodiments, CD8 T cells in a subject have enhanced priming, activation, proliferation, and / or cytolytic activity relative to prior administration of a compound of the invention, or a pharmaceutically acceptable salt, prodrug, metabolite, or derivative thereof. In some embodiments, CD8 T cell priming is characterized by elevated CD44 expression and / or enhanced cytolytic activity in CD8 T cells. In some embodiments, CD8 T cell activation is characterized by increased γ-IFN expression and / or increased cytolytic activity in CD8 T cells. + In some embodiments, the CD8 T cells are antigen-specific T cells.

[0179] In some embodiments, antigen-presenting cells in a subject have enhanced maturation and activation compared to before administration of a compound of the present invention, or a pharmaceutically acceptable salt, prodrug, metabolite, or derivative thereof. In some embodiments, the antigen-presenting cells are dendritic cells. In some embodiments, the maturation of antigen-presenting cells is enhanced by CD83 expression. + In some embodiments, activation of antigen-presenting cells is characterized by increased frequency of dendritic cells. In some embodiments, activation of antigen-presenting cells is characterized by increased expression of CD80 and CD86 on dendritic cells.

[0180] In some embodiments, the serum levels of the cytokine IL-10 and / or the chemokine IL-8, which are human homologs of mouse KC, in the subject are reduced compared to before administration of the compound of Formula I or Ia, or a pharmaceutically acceptable salt, prodrug, metabolite, or derivative thereof.

[0181] TCR engagement leads to activation of HPK1, which functions as a negative regulator of the TCR-induced AP-1 response pathway. HPK1 is thought to negatively regulate T cell activation by phosphorylating SLP76 at Ser376 (Di Bartolo et al. (2007) JEM 204:681-691) and Gads at Thr254, thereby reducing the persistence of signaling microclusters. This leads to the recruitment of 14-3-3 proteins that bind to phosphorylated SLP76 and Gads, resulting in the release of the SLP76-Gads-14-3-3 complex from LAT-containing microclusters, leading to T cell dysfunction, including anergy and exhaustion (Lasserre et al. (2011) J Cell Biol 195(5):839-853).

[0182] In some embodiments, administration of a compound of the invention, or a pharmaceutically acceptable salt, prodrug, metabolite, or derivative thereof, to a subject results in enhanced T cell function.

[0183] Thus, the disclosed compounds of the present invention or pharmaceutically acceptable salts, prodrugs, metabolites, or derivatives thereof are useful in treating T cell dysfunction disorders. A "T cell dysfunction disorder" is a disorder or condition of T cells characterized by reduced responsiveness to antigenic stimulation. In certain embodiments, the T cell dysfunction disorder is a disorder specifically associated with increased kinase activity of HPK1. In another embodiment, the T cell dysfunction disorder is one in which T cells are anergic or have a reduced ability to secrete cytokines, proliferate, or achieve cytolytic activity. In certain aspects, the reduced responsiveness results in ineffective control of pathogens or tumors expressing immunogens. Examples of T cell dysfunction disorders characterized by T cell dysfunction include unresolved acute infections, chronic infections, and tumor immunity.

[0184] Thus, the compounds of the present disclosure can be used in the treatment of conditions where enhanced immunogenicity is desirable, such as increasing tumor immunogenicity for the treatment of cancer.

[0185] The term "dysfunction" in the context of immune dysfunction refers to a state of reduced immune responsiveness to antigenic stimulation. This term includes the general components of both exhaustion and / or anergy, in which antigen recognition can occur, but the subsequent immune response is ineffective in controlling infection or tumor growth.

[0186] As used herein, the term "dysfunctional" also refers to refractoriness or non-responsiveness to antigen recognition, specifically including impaired ability to translate antigen recognition into downstream T cell effector functions such as proliferation, cytokine production (e.g., IL-2, γ-IFN) and / or target cell killing.

[0187] The term "anergy" refers to the incomplete or insufficient signal delivered via the T cell receptor (e.g., intracellular Ca in the absence of ras activation). +2T cell anergy refers to a state of non-responsiveness to antigenic stimulation resulting from stimulation with antigen in the absence of costimulation, rendering the cells refractory to subsequent activation by antigen even in the context of costimulation. The unresponsive state can often be reversed by the presence of interleukin-2. Anergic T cells do not undergo clonal expansion and / or acquire effector function.

[0188] The term "exhaustion" refers to T cell exhaustion, a state of T cell dysfunction resulting from persistent TCR signaling, which occurs during many chronic infections and cancers. It is distinguished from anergy in that it results from persistent signaling rather than incomplete or defective signaling. It is defined by insufficient effector function, persistent expression of inhibitory receptors, and a transcriptional state that differs from functional effector or memory T cells. Exhaustion prevents optimal control of infection and tumors. Exhaustion can result from both extrinsic negative regulatory pathways (e.g., immunomodulatory cytokines) and endogenous negative regulatory (costimulatory) pathways (PD-1, B7-H3, B7-H4, etc.).

[0189] "Immunogenicity" refers to the ability of a particular substance to induce an immune response. Tumors are immunogenic, and enhancing tumor immunogenicity aids in the clearance of tumor cells by the immune response.

[0190] "Enhancing T cell function" means inducing, causing, or stimulating T cells to have sustained or amplified biological function, or to regenerate or reactivate exhausted or inactive T cells. Examples of enhancing T cell function include increased cytokine secretion (e.g., gamma-interferon, IL-2, IL-12, and TNFα), increased proliferation, increased antigen responsiveness to such levels prior to the intervention (e.g., viral, pathogen, or tumor clearance), and increased production of effector granules by CD8 T cells, such as granzyme B. In one embodiment, the level of enhancement is at least 50%, alternatively 60%, 70%, 80%, 90%, 100%, 120%, 150%, or 200%. Methods for measuring this enhancement are known to those skilled in the art.

[0191] "Tumor immunity" refers to the process by which tumors evade immune recognition and clearance. Thus, as a therapeutic concept, tumor immunity is "treated" when such evasion is attenuated and the tumor is recognized and attacked by the immune system. Examples of tumor recognition include tumor binding, tumor shrinkage, and tumor clearance.

[0192] The present disclosure provides methods for modulating (eg, inhibiting) HPKl activity, the methods comprising administering to a patient a compound provided herein, or a pharmaceutically acceptable salt thereof.

[0193] In one aspect, provided herein is a method for treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound of the invention, or a pharmaceutically acceptable salt, prodrug, metabolite, or derivative thereof.

[0194] In the methods described herein, a compound of the present invention or a pharmaceutical composition thereof is administered to a subject with cancer.

[0195] In certain embodiments, the presently disclosed subject matter is directed to a method for treating an HPK1-dependent disorder, the method comprising administering to a subject in need thereof an effective amount of a compound of the invention or a pharmaceutical composition described herein. In certain aspects of this embodiment, the HPK1-dependent disorder is cancer. In certain aspects of this embodiment, the cancer comprises at least one cancer selected from the group consisting of colorectal cancer, melanoma, non-small cell lung cancer, ovarian cancer, breast cancer, pancreatic cancer, hematological malignancies, and renal cell carcinoma. In certain aspects of this embodiment, the cancer has an elevated level of T cell infiltration. In certain aspects of this embodiment, the cancer cells in the subject selectively have elevated expression of MHC class I antigens compared to before administration of the compound or composition.

[0196] In some embodiments, the presently disclosed subject matter is directed to a method for treating chronic viral infections. In some embodiments, the presently disclosed subject matter is directed to the use of HPK1 inhibitors as adjuvant therapy to increase the efficacy of vaccination.

[0197] In some embodiments, the present invention provides a pharmaceutical composition comprising an effective amount of a compound of the present invention, or a pharmaceutically acceptable salt, hydrate, solvate, or prodrug thereof, and a pharmaceutically acceptable carrier.

[0198] In certain aspects, the present invention provides methods for treating cell proliferative disorders, including cancer, benign papillomas, gestational trophoblastic disease, and benign neoplastic diseases such as cutaneous papillomas (warts) and genital papillomas.

[0199] In one aspect, the invention provides a method of treating a cell proliferative disorder in a subject, comprising administering to the subject a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt, hydrate, solvate, or prodrug thereof.

[0200] In certain embodiments, the cell proliferative disorder is cancer.

[0201] Examples of cancers that can be treated using the compounds of the present disclosure include bone cancer, pancreatic cancer, skin cancer, cancer of the head and neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, cancer of the fallopian tubes, cancer of the lining of the uterus, endometrial cancer, cancer of the cervix, cancer of the vagina, cancer of the vulva, Hodgkin's lymphoma, non-Hodgkin's lymphoma, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, acute myeloid leukemia, These include, but are not limited to, chronic or acute leukemias, including chronic myeloid leukemia, acute lymphoblastic leukemia, and chronic lymphocytic leukemia; solid tumors of childhood; lymphocytic lymphoma; cancer of the bladder; cancer of the kidney or urethra; cancer of the renal pelvis; neoplasms of the central nervous system (CNS); primary CNS lymphoma; tumor angiogenesis; spinal axis tumors; brain stem glioma; pituitary adenoma; Kaposi's sarcoma; epidermoid carcinoma; squamous cell carcinoma; T-cell lymphoma; environmentally induced cancers, including asbestos-induced cancers; and combinations of such cancers.

[0202] In some embodiments, cancers treatable using compounds of the present disclosure include, but are not limited to, solid tumors (e.g., prostate cancer, colon cancer, esophageal cancer, endometrial cancer, ovarian cancer, uterine cancer, kidney cancer, liver cancer, pancreatic cancer, stomach cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, sarcoma, bladder cancer, etc.), hematological cancers (e.g., leukemias such as lymphoma, acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), DLBCL, mantle cell lymphoma, non-Hodgkin's lymphoma (including relapsed or refractory NHL and relapsed follicular), Hodgkin's lymphoma, or multiple myeloma), and combinations of such cancers.

[0203] In certain embodiments, the cancer is brain cancer, leukemia, skin cancer, prostate cancer, thyroid cancer, colon cancer, lung cancer, or sarcoma, hi another embodiment, the cancer is selected from the group consisting of glioma, glioblastoma multiforme, paraganglioma, supratentorial primitive neuroectodermal tumor, acute myeloid leukemia, myelodysplastic syndrome, chronic myeloid leukemia, melanoma, breast, prostate, thyroid, colon, lung, central chondrosarcoma, central and periosteal chondroma tumor, fibrosarcoma, and cholangiocarcinoma.

[0204] In certain embodiments, the cancer is selected from brain and spinal cord cancer, head and neck cancer, leukemia and blood cancer, skin cancer, reproductive system cancer, digestive system cancer, liver and bile duct cancer, kidney and bladder cancer, bone cancer, lung cancer, malignant mesothelioma, sarcoma, lymphoma, adenocarcinoma, thyroid cancer, cardiac tumors, germ cell tumors, malignant neuroendocrine (carcinoid) tumors, midline duct carcinoma, and cancer of unknown primary origin (cancer in which metastases are found but the site of original cancer is unknown). In certain embodiments, the cancer is in an adult patient, and in additional embodiments, the cancer is in a pediatric patient. In certain embodiments, the cancer is AIDS-related.

[0205] In further embodiments, the cancer is selected from brain cancer and spinal cord cancer. In certain embodiments, the cancer is selected from the group consisting of anaplastic astrocytoma, glioblastoma, astrocytoma, and nasal neuroblastoma (olfactory neuroblastoma). In certain embodiments, the brain cancer is selected from the group consisting of astrocytic tumors (e.g., pilocytic astrocytoma, subependymal giant cell astrocytoma, diffuse astrocytoma, pleomorphic xanthoastrocytoma, anaplastic astrocytoma, astrocytoma, giant cell glioblastoma, glioblastoma, secondary glioblastoma, primary adult glioblastoma, and primary pediatric glioblastoma), oligodendroglial tumors (e.g., oligodendroglioma and anaplastic oligodendroglioma), oligoastrocytic tumors (e.g., glioma ... In certain embodiments, the brain cancer is selected from the group consisting of glioma, glioblastoma multiforme, paraganglioma, and supratentorial primitive neuroectodermal tumor (sPNET).

[0206] In certain embodiments, the cancer is selected from nasopharyngeal cancer, cancer of the nasal cavity and paranasal sinuses, hypopharyngeal cancer, oral cancer (e.g., squamous cell carcinoma, lymphoma, and sarcoma), lip cancer, oropharyngeal cancer, salivary gland tumors, cancer of the larynx (e.g., laryngeal squamous cell carcinoma, rhabdomyosarcoma), and cancer of the head and neck, including eye cancer and ocular cancer. In certain embodiments, the eye cancer is selected from the group consisting of intraocular melanoma and retinoblastoma.

[0207] In certain embodiments, the cancer is selected from leukemia and hematological cancer. In certain embodiments, the cancer is selected from the group consisting of myeloproliferative neoplasms, myelodysplastic syndromes, myelodysplastic / myeloproliferative neoplasms, acute myeloid leukemia (AML), myelodysplastic syndromes (MDS), chronic myeloid leukemia (CML), myeloproliferative neoplasms (MPN), post-MPN AML, post-MDS AML, del(5q)-associated high-risk MDS or AML, blastic phase chronic myeloid leukemia, angioimmunoblastic lymphoma, acute lymphocytic leukemia, Langerhans cell histiocytosis, hairy cell leukemia, and plasma cell neoplasms, including plasmacytoma and multiple myeloma. The leukemias referred to herein can be acute or chronic.

[0208] In certain embodiments, the cancer is selected from skin cancer, hi certain embodiments, the skin cancer is selected from the group consisting of melanoma, squamous cell carcinoma, and basal cell carcinoma.

[0209] In certain embodiments, the cancer is selected from cancers of the reproductive system. In certain embodiments, the cancer is selected from the group consisting of breast cancer, cervical cancer, vaginal cancer, ovarian cancer, prostate cancer, penile cancer, and testicular cancer. In certain examples of these embodiments, the cancer is breast cancer selected from the group consisting of ductal carcinoma and phyllodes tumor. In certain examples of these embodiments, the breast cancer can be male breast cancer or female breast cancer. In certain examples of these embodiments, the cancer is cervical cancer selected from the group consisting of squamous cell carcinoma and adenocarcinoma. In certain examples of these embodiments, the cancer is ovarian cancer selected from the group consisting of epithelial cancers.

[0210] In certain embodiments, the cancer is selected from a cancer of the digestive system. In certain embodiments, the cancer is selected from the group consisting of esophageal cancer, gastric cancer (also known as stomach cancer), gastrointestinal carcinoid tumor, pancreatic cancer, gallbladder cancer, colorectal cancer, and anal cancer. In exemplary of these embodiments, the cancer is selected from the group consisting of esophageal squamous cell carcinoma, esophageal adenocarcinoma, gastric adenocarcinoma, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, gastric lymphoma, gastrointestinal lymphoma, solid pseudopapillary tumor of the pancreas, pancreatoblastoma, pancreatic islet cell tumor, pancreatic cancer including acinar cell carcinoma and ductal adenocarcinoma, gallbladder adenocarcinoma, colorectal adenocarcinoma, and anal squamous cell carcinoma.

[0211] In certain embodiments, the cancer is selected from liver cancer and cholangiocarcinoma. In certain embodiments, the cancer is liver cancer (hepatocellular carcinoma). In certain embodiments, the cancer is bile duct cancer (cholangiocarcinoma), and in exemplary of these embodiments, the cholangiocarcinoma is selected from the group consisting of intrahepatic cholangiocarcinoma and extrahepatic cholangiocarcinoma.

[0212] In certain embodiments, the cancer is selected from kidney cancer and bladder cancer. In certain embodiments, the cancer is kidney cancer selected from the group consisting of renal cell carcinoma, Wilms' tumor, and transitional cell carcinoma. In certain embodiments, the cancer is bladder cancer selected from the group consisting of urothelial carcinoma (transitional cell carcinoma), squamous cell carcinoma, and adenocarcinoma.

[0213] In certain embodiments, the cancer is selected from bone cancer, hi certain embodiments, the bone cancer is selected from the group consisting of osteosarcoma, malignant fibrous histiocytoma of bone, Ewing's sarcoma, and chordoma.

[0214] In certain embodiments, the cancer is selected from lung cancer. In certain embodiments, the lung cancer is selected from the group consisting of non-small cell lung cancer, small cell lung cancer, bronchial tumor, and pleuropulmonary blastoma.

[0215] In certain embodiments, the cancer is selected from the group consisting of malignant mesothelioma, epithelioid mesothelioma, and sarcomatoid.

[0216] In certain embodiments, the cancer is selected from a sarcoma, hi certain embodiments, the sarcoma is selected from the group consisting of central chondrosarcoma, central and periosteal chondroma, fibrosarcoma, clear cell sarcoma of the tendon sheath, and Kaposi's sarcoma.

[0217] In certain embodiments, the cancer is selected from the group consisting of Hodgkin's lymphoma (e.g., Reed-Sternberg cell), non-Hodgkin's lymphoma (e.g., diffuse large B-cell lymphoma, follicular lymphoma, mycosis fungoides, Sézary syndrome, primary central nervous system lymphoma), cutaneous T-cell lymphoma, and primary central nervous system lymphoma.

[0218] In certain embodiments, the cancer is selected from an adenocarcinoma, hi certain embodiments, the cancer is selected from the group consisting of an adrenocortical carcinoma, a pheochromocytoma, a paraganglioma, a pituitary tumor, a thymoma, and a thymic carcinoma.

[0219] In certain embodiments, the cancer is selected from thyroid cancer, hi certain embodiments, the thyroid cancer is selected from the group consisting of medullary thyroid cancer, papillary thyroid cancer, and follicular thyroid cancer.

[0220] In certain embodiments, the cancer is selected from a germ cell tumor. In certain embodiments, the cancer is selected from the group consisting of malignant extracranial germ cell tumor and malignant extragonadal germ cell tumor. In certain examples of these embodiments, the malignant extragonadal germ cell tumor is selected from the group consisting of nonseminoma and seminoma.

[0221] In certain embodiments, the cancer is selected from a cardiac tumor, hi certain embodiments, the cardiac tumor is selected from the group consisting of malignant teratoma, lymphoma, rhabdomyoma, angiosarcoma, chondrosarcoma, infantile fibrosarcoma, and synovial sarcoma.

[0222] In certain embodiments, the cell proliferative disorder is selected from benign papilloma, benign neoplastic disease, and gestational trophoblastic disease. In certain embodiments, the benign neoplastic disease is selected from cutaneous papilloma (warts) and genital papilloma. In certain embodiments, the gestational trophoblastic disease is selected from the group consisting of hydatidiform mole and gestational trophoblastic neoplasm (e.g., invasive mole, choriocarcinoma, placental site trophoblastic tumor, and epithelioid trophoblastic tumor).

[0223] In some embodiments, the subject has melanoma. The melanoma can be at an early or late stage. In some embodiments, the subject has colon cancer. The colon cancer can be at an early or late stage. In some embodiments, the subject has non-small cell lung cancer. The non-small cell lung cancer can be at an early or late stage. In some embodiments, the subject has pancreatic cancer. The pancreatic cancer can be at an early or late stage. In some embodiments, the subject has a hematological malignancy. The hematological malignancy can be at an early or late stage. In some embodiments, the subject has ovarian cancer. The ovarian cancer can be at an early or late stage. In some embodiments, the subject has breast cancer. The breast cancer can be at an early or late stage. In some embodiments, the subject has renal cell carcinoma. The renal cell carcinoma can be at an early or late stage. In some embodiments, the cancer has elevated levels of T cell infiltration.

[0224] In some embodiments, cancers treatable with the compounds of the present disclosure include melanoma (e.g., metastatic malignant melanoma), kidney cancer (e.g., clear cell carcinoma), prostate cancer (e.g., hormone-refractory prostate adenocarcinoma), breast cancer, triple-negative breast cancer, colon cancer, and lung cancer (e.g., non-small cell lung cancer and small cell lung cancer). Additionally, the present disclosure includes resistant or recurrent malignancies whose growth can be inhibited using the compounds of the present disclosure.

[0225] In some embodiments, diseases and indications treatable using compounds of the present disclosure include, but are not limited to, hematological cancers, sarcomas, lung cancers, gastrointestinal cancers, genitourinary cancers, liver cancers, bone cancers, nervous system cancers, gynecological cancers, and skin cancers.

[0226] Exemplary hematological cancers include lymphomas and leukemias such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma, non-Hodgkin's lymphoma (including relapsed or refractory NHL and relapsed follicular), Hodgkin's lymphoma, myeloproliferative disorders (e.g., primary myelofibrosis (PMF), polycythemia vera (PV), essential thrombocythemia (ET), myelodysplastic syndromes (MDS), T-cell acute lymphoblastic lymphoma (T-ALL), multiple myeloma, cutaneous T-cell lymphoma, Waldenstrom's macroglobulinemia, hairy cell lymphoma, chronic myeloid lymphoma, and Burkitt's lymphoma.

[0227] Exemplary sarcomas include chondrosarcoma, Ewing's sarcoma, osteosarcoma, rhabdomyosarcoma, angiosarcoma, fibrosarcoma, liposarcoma, myxoma, rhabdomyoma, rhabdomyosarcoma, fibroma, lipoma, hamartoma, and teratoma.

[0228] Exemplary lung cancers include non-small cell lung cancer (NSCLC), small cell lung cancer, bronchial carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchial) carcinoma, bronchial adenoma, chondroitin hamartoma, and mesothelioma.

[0229] Exemplary gastrointestinal cancers include esophageal cancer (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), gastric cancer (carcinoma, lymphoma, leiomyosarcoma), pancreatic cancer (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, vipoma), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), colon (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma), and colorectal cancer.

[0230] Exemplary genitourinary cancers include cancer of the kidney (adenocarcinoma, Wilms' tumor [nephroblastoma]), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), and testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell tumor, fibroma, fibroadenoma, adenomatous tumor, lipoma).

[0231] Exemplary liver cancers include hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, and hemangioma.

[0232] Exemplary bone cancers include, for example, osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell chordoma, osteochondroma (osteochondral exostosis), benign chondroma, chondroblastoma, chondromyxoid fibroma, osteoid osteoma, and giant cell tumor.

[0233] Exemplary nervous system cancers include cancers of the skull (osteoma, hemangioma, granuloma, xanthomas, osteitis deformans), cancers of the meninges (meningioma, meningeal sarcoma, gliomatosis), cancers of the brain (astrocytoma, medulloblastoma, glioma, ependymoma, embryonal tumor (pinealoma), glioblastoma, glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), and cancers of the spinal cord (neurofibroma, meningioma, glioma, sarcoma), as well as neuroblastoma and Lhermitte-Duclos disease.

[0234] Exemplary gynecological cancers include cancer of the uterus (endometrial carcinoma), cancer of the cervix (cervical carcinoma, preneoplastic cervical dysplasia), cancer of the ovaries (ovarian carcinoma (serous cystadenoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosathecocytoma, Sertoli-Leydig cell tumor, dysgerminoma, malignant teratoma), cancer of the vulva (squamous cell carcinoma, carcinoma in situ, adenocarcinoma, fibrosarcoma, melanoma), cancer of the vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma)), and cancer of the fallopian tubes (carcinoma).

[0235] Exemplary skin cancers include melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, Merkel cell skin cancer, dysplastic nevi, lipoma, hemangioma, dermatofibroma, and keloids. In some embodiments, diseases and indications treatable using the compounds of the present disclosure include, but are not limited to, sickle cell disease (e.g., sickle cell anemia), triple-negative breast cancer (TNBC), myelodysplastic syndrome, testicular cancer, bile duct cancer, esophageal cancer, and urothelial cancer.

[0236] Exemplary head and neck cancers include glioblastoma, melanoma, rhabdomyosarcoma, lymphosarcoma, osteosarcoma, squamous cell carcinoma, adenocarcinoma, oral cavity cancer, laryngeal cancer, nasopharyngeal cancer, nasal cavity and paranasal sinus cancer, thyroid cancer, and parathyroid cancer.

[0237] In some embodiments, HPK1 inhibitors can be used to treat tumors that produce PGE2 (e.g., Cox-2-overexpressing tumors) and / or adenosine (CD73 and CD39-overexpressing tumors). Cox-2 overexpression has been detected in several tumors, such as colon cancer, breast cancer, pancreatic cancer, and lung cancer, where it correlates with poor prognosis. COX-2 overexpression has been reported in hematological cancer models such as RAJI (Burkitt's lymphoma) and U937 (acute promonocytic leukemia), as well as in blast cells from patients. CD73 is upregulated in various human carcinomas, including colon, lung, pancreatic, and ovarian carcinomas. Importantly, high CD73 expression levels are associated with tumor neovascularization, invasiveness, and metastasis, and shorter patient survival in breast cancer.

[0238] In some embodiments, the compounds of the invention are useful for preventing or reducing the risk of developing any of the diseases mentioned herein, for example, for preventing or reducing the risk of developing a disease, condition, or disorder in individuals who may be predisposed to the disease, condition, or disorder but who have not yet experienced or exhibited the pathology or symptoms of the disease.

[0239] The compounds of the present disclosure can be administered in any suitable manner known in the art, hi some embodiments, the compounds of the present invention or pharmaceutically acceptable salts, prodrugs, metabolites, or derivatives thereof are administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intracerebroventricularly, intratumorally, or intranasally.

[0240] In some embodiments, the HPK1 antagonist is administered continuously. In other embodiments, the HPK1 antagonist is administered intermittently. Moreover, treatment of a subject with an effective amount of an HPK1 antagonist can include a single treatment or can include a series of treatments.

[0241] It will be understood that the appropriate dose of the active compound will depend on several factors within the knowledge of a physician or veterinarian of ordinary skill in the art. The dose of the active compound will vary depending on, for example, the age, weight, general health, sex, and diet of the subject, the time of administration, the route of administration, the rate of excretion, and any drug combinations.

[0242] It will also be understood that the effective dosage of the compounds of the invention, or pharmaceutically acceptable salts, prodrugs, metabolites, or derivatives thereof, used for treatment may increase or decrease over the course of a particular treatment. Modifications in dosage may arise and become apparent from the results of diagnostic assays.

[0243] In some embodiments, the HPK1 antagonist is administered to a subject at a dose of about 0.001 μg / μkg to about 1000 mg / kg, including, but not limited to, about 0.001 μg / kg, 0.01 μg / kg, 0.05 μg / kg, 0.1 μg / kg, 0.5 μg / kg, 1 μg / kg, 10 μg / kg, 25 μg / kg, 50 μg / kg, 100 μg / kg, 250 μg / kg, 500 μg / kg, 1 mg / kg, 5 mg / kg, 10 mg / kg, 25 mg / kg, 50 mg / kg, 100 mg / kg, and 200 mg / kg.

[0244] In the methods described herein, the methods can further include administering a chemotherapeutic agent to the subject. In certain aspects of this embodiment, the chemotherapeutic agent is administered to the subject simultaneously with the compound or composition. In certain aspects of this embodiment, the chemotherapeutic agent is administered to the subject prior to administration of the compound or composition. In certain aspects of this embodiment, the chemotherapeutic agent is administered to the subject after administration of the compound or composition.

[0245] As used herein, the terms "treatment," "treat," and "treating" refer to reversing, alleviating, delaying the onset of, or inhibiting the progression of a disease or disorder described herein, or one or more symptoms thereof. In some embodiments, treatment may be administered after one or more symptoms have developed. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and / or in light of genetic or other susceptibility factors). Treatment may also be continued after symptoms have resolved, e.g., to prevent or delay recurrence.

[0246] The term "administration" or "administering" includes routes of introducing a compound into a subject to perform its intended function. Examples of routes of administration that may be used include injection (subcutaneous, intravenous, parenteral, intraperitoneal, intrathecal), topical, oral, inhalation, rectal, and transdermal.

[0247] The term "effective amount" includes an amount effective, at dosages and for periods of time necessary, to achieve the desired result. The effective amount of a compound may vary depending on factors such as the disease state, age, and weight of the subject, and the ability of the compound to elicit a desired response in the subject. Dosage regimens may be adjusted to provide the optimal therapeutic response.

[0248] As used herein, the phrases "systemic administration," "administered systemically," "peripheral administration," and "administered peripherally" refer to the administration of a compound, drug, or other substance so that it enters the patient's system and is therefore subject to metabolic and other similar processes.

[0249] The phrase "therapeutically effective amount" refers to an amount of a compound of the present invention that (i) treats or prevents a particular disease, condition, or disorder, (ii) attenuates, ameliorate, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition, or disorder described herein. In the case of cancer, a therapeutically effective amount may reduce the number of cancer cells, reduce tumor size, inhibit (i.e., slow to some extent, and preferably stop) cancer cell invasion into peripheral organs, inhibit (i.e., slow to some extent, and preferably stop) tumor metastasis, inhibit tumor growth to some extent, and / or alleviate to some extent one or more symptoms associated with cancer. To the extent that the agent can prevent growth and / or kill existing cancer cells, it may be cytostatic and / or cytotoxic. For cancer therapy, efficacy can be measured, for example, by assessing the time to disease progression (TTP) and / or determining the response rate (RR).

[0250] The term "subject" refers to animals such as mammals, including, but not limited to, primates (e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, etc. In certain embodiments, the subject is a human.

[0251] Combination therapy Depending on the particular condition, or disease, being treated, additional therapeutic agents, which are normally administered to treat that condition, may be administered in combination with the compounds and compositions of this invention. As used herein, additional therapeutic agents that are normally administered to treat a particular disease or condition are known as "appropriate for the disease, or condition, being treated."

[0252] In certain embodiments, a provided combination, or composition thereof, is administered in combination with another therapeutic agent.

[0253] Examples of drugs that may also be combined with the agents of the invention include, but are not limited to, treatments for Alzheimer's disease such as Aricept® and Excelon®; treatments for HIV such as ritonavir; treatments for Parkinson's disease such as L-DOPA / carbidopa, entacapone, lopinrol, pramipexole, bromocriptine, pergolide, trihexefendil, and amantadine; drugs for treating multiple sclerosis (MS) such as beta interferons (e.g., Avonex® and Rebif®), Copaxone®, and mitoxantrone; treatments for asthma such as albuterol and Singulair®; drugs for treating schizophrenia such as Zyprexa, Risperdal, Seroquel, and haloperidol; corticosteroids, TNF blockers, IL-1 Anti-inflammatory drugs such as RA, azathioprine, cyclophosphamide, and sulfasalazine; immunomodulators and immunosuppressants such as cyclosporine, tacrolimus, rapamycin, mycophenolate mofetil, interferon, corticosteroids, cyclophosphamide, azathioprine, and sulfasalazine; neurotrophic factors such as acetylcholinesterase inhibitors, MAO inhibitors, interferon, antispasmodics, ion channel blockers, riluzole, and antiparkinsonian drugs; beta-blockers, ACE inhibitors, diuretics, nitrates, calcium channel blockers drugs for treating cardiovascular disease, such as drugs and statins; drugs for treating liver disease, such as corticosteroids, cholestyramine, interferons, and antivirals; drugs for treating blood disorders, such as corticosteroids, anti-leukemia drugs, and growth factors; drugs that prolong or improve pharmacokinetics, such as cytochrome P450 inhibitors (i.e., inhibitors of metabolic breakdown) and CYP3A4 inhibitors (e.g., ketokenozole and ritonavir); and drugs for treating immune deficiency disorders, such as gamma globulins.

[0254] In certain embodiments, the combination therapy of the invention, or a pharmaceutically acceptable composition thereof, is administered in combination with a monoclonal antibody or siRNA therapeutic agent.

[0255] These additional agents can be administered separately from the combination therapy provided as part of a multiple dose regimen. Alternatively, these agents can be part of a single dosage form, mixed together with the compound of the present invention in a single composition. When administered as part of a multiple dose regimen, the two active agents can be given simultaneously, sequentially, or within a certain period of each other, usually within 5 hours of each other.

[0256] As used herein, the terms "combination," "in combination," and related terms refer to simultaneous or sequential administration of therapeutic agents according to the present invention. For example, a combination of the present invention may be administered with another therapeutic agent, either simultaneously or sequentially, in separate unit dosage forms or together in a single unit dosage form.

[0257] The amount of additional therapeutic agent present in the compositions of the invention is no more than the amount that would normally be administered in a composition comprising that therapeutic agent as the only active agent. Preferably, the amount of additional therapeutic agent in the compositions of the present disclosure ranges from about 50% to 100% of the amount that would normally be present in a composition comprising that agent as the only therapeutically active agent.

[0258] In one embodiment, the present invention provides a composition comprising a compound of Formula I and one or more additional therapeutic agents. The therapeutic agents may be administered together with the compound of Formula I, or may be administered before or after the administration of the compound of Formula I. Suitable therapeutic agents are described in more detail below. In certain embodiments, the compound of Formula I may be administered up to 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, or 18 hours before the therapeutic agent. In other embodiments, the compound of formula I may be administered up to 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, or 18 hours after the therapeutic agent.

[0259] In another embodiment, the present invention provides a method of treating an inflammatory disease, disorder, or condition by administering to a patient in need thereof a compound of Formula I and one or more additional therapeutic agents. Such additional therapeutic agents may be small molecules or recombinant biological agents, for example, nonsteroidal anti-inflammatory drugs (NSAIDS) such as acetaminophen, aspirin, ibuprofen, naproxen, etodolac (Lodine®), and celecoxib; colchicine (Colcrys®), corticosteroids such as prednisone, prednisolone, methylprednisolone, hydrocortisone; probenecid; allopurinol; febuxostat (Uloric®); Antimalarials such as sulfasalazine (Azulfidine®), hydroxychloroquine (Plaquenil®) and chloroquine (Aralen®), methotrexate (Rheumatrex®), gold salts such as gold thioglucose (Solganal®), gold thiomalate (Myochrysine®), and auranofin (Ridaura®), D-penicillamine (Depen® or Cuprimine®), ®), azathioprine (Imuran®), cyclophosphamide (Cytoxan®), chlorambucil (Leukeran®), cyclosporine (Sandimmune®), leflunomide (Arava®), as well as etanercept (Enbrel®), infliximab (Remicade®), golimumab (Simponi®), certolizumab pegol (Cimzia®), and adalimumab. "Anti-TNF" agents such as rilonacept (Humira®), "anti-IL-1" agents such as anakinra (Kineret®) and rilonacept (Arcalyst®), anti-Jak inhibitors such as canakinumab (Ilaris®) and tofacitinib, antibodies such as rituximab (Rituxan®), "anti-T cell" agents such as abatacept (Orencia®), "anti-IL-6" agents such as tocilizumab (Actemra®), diclofenac, cortisone,Hyaluronic acid (Synvisc® or Hyalgan®), monoclonal antibodies such as tanezumab, anticoagulants such as heparin (Calcinparine® or Liquaemin®) and warfarin (Coumadin®), antidiarrheal drugs such as diphenoxylate (Lomotil®) and loperamide (Imodium®), bile acid binders such as cholestyramine, alosetron (Lotronex®), lubiprostone (Amitiza®), magnesium Shea milk, polyethylene glycol (MiraLax®), laxatives such as Dulcolax®, Correctol®, and Senokot®, anticholinergics or antispasmodics such as dicyclomine (Bentyl®), Singulair®, albuterol (Ventolin® HFA, Proventil® HFA), levalbuterol (Xopenex®), metaproterenol (Alupent®), pirbuterol acetate (Maxair®), beta-2 agonists such as terbutaline sulfate (Brethaire®), salmeterol xinafoate (Serevent®) and formoterol (Foradil®), anticholinergics such as ipratropium bromide (Atrovent®) and tiotropium (Spiriva®), beclomethasone dipropionate (Beclovent®, Qvar®, and Vanceril®), triamcinolone acetonide (Azmacort®), inhaled corticosteroids such as mometasone (Asthmanex®), budesonide (Pulmocort®), and flunisolide (Aerobid®), methylxanthines such as Afviar®, Symbicort®, Dulera®, cromolyn sodium (Intal®), theophylline (Theo-Dur®, Theolair®, Slo-bid®, Uniphyl®, Theo-24®) and aminophylline,IgE antibodies such as omalizumab (Xolair®), zidovudine (Retrovir®), abacavir (Ziagen®), abacavir / lamivudine (Epzicom®), abacavir / lamivudine / zidovudine (Trizivir®), didanosine (Videx®), emtricitabine (Emtriva®), lamivudine (Epivir®), lamivudine / zidovudine (Combivir®), stavudine (Zerit®), )), and zalcitabine (Hivid®), non-nucleoside reverse transcriptase inhibitors such as delavirdine (Rescriptor®), efavirenz (Sustiva®), nevirapine (Viramune®), and etravirine (Intelence®), nucleotide reverse transcriptase inhibitors such as tenofovir (Viread®), amprenavir (Agenerase®), atazanavir (Azadafenib), and rivaroxaban (Renavir®). protease inhibitors such as fluprenavir (Reyataz®), darunavir (Prezista®), fosamprenavir (Lexiva®), indinavir (Crixivan®), lopinavir and ritonavir (Kaletra®), nelfinavir (Viracept®), ritonavir (Norvir®), saquinavir (Fortovase® or Invirase®), and tipranavir (Aptivus®), enfuvir entry inhibitors such as rivaroxaban (Fuzeon®) and maraviroc (Selzentry®), integrase inhibitors such as raltegravir (Isentress®), doxorubicin (Hydrodaunorubicin®), vincristine (Oncovin®), bortezomib (Velcade®), and dexamethasone (Decadron®) in combination with lenalidomide (Revlimid®), or any combination thereof.

[0260] In another embodiment, the present invention provides a method of treating rheumatoid arthritis, comprising administering to a patient in need thereof a compound of Formula I and nonsteroidal anti-inflammatory drugs (NSAIDS) such as aspirin, ibuprofen, naproxen, etodolac (Lodine®), and celecoxib; corticosteroids such as prednisone, prednisolone, methylprednisolone, hydrocortisone; antimalarials such as sulfasalazine (Azulfidine®), hydroxychloroquine (Plaquenil®), and chloroquine (Aralen®); gold salts such as methotrexate (Rheumatrex®), gold thioglucose (Solganal®), gold thiomalate (Myochrysine®), and auranofin (Ridaura®); D-penicillamine (Depen®) or Cuprimine®. )), azathioprine (Imuran®), cyclophosphamide (Cytoxan®), chlorambucil (Leukeran®), cyclosporine (Sandimmune®), leflunomide (Arava®), as well as etanercept (Enbrel®), infliximab (Remicade®), golimumab (Simponi®), certolizumab pegol (Cimzia®), and one or more additional therapeutic agents selected from "anti-TNF" agents such as adalimumab (Humira®), "anti-IL-1" agents such as anakinra (Kineret®) and rilonacept (Arcalyst®), "anti-T cell" agents such as abatacept (Orencia®), and "anti-IL-6" agents such as tocilizumab (Actemra®).

[0261] In some embodiments, the present invention provides a method of treating osteoarthritis, comprising administering to a patient in need thereof a compound of Formula I and one or more additional therapeutic agents selected from acetaminophen, aspirin, ibuprofen, naproxen, etodolac (Lodine®), and nonsteroidal anti-inflammatory drugs (NSAIDS) such as celecoxib, diclofenac, cortisone, hyaluronic acid (Synvisc® and Hyalgan®), and monoclonal antibodies such as tanezumab.

[0262] In some embodiments, the present invention provides a method of treating cutaneous lupus erythematosus or systemic lupus erythematosus, comprising administering to a patient in need thereof a compound of Formula I and one or more additional therapeutic agents selected from nonsteroidal anti-inflammatory drugs (NSAIDS) such as acetaminophen, aspirin, ibuprofen, naproxen, etodolac (Lodine®), and celecoxib; corticosteroids such as prednisone, prednisolone, methylprednisolone, hydrocortisone; antimalarials such as hydroxychloroquine (Plaquenil®) and chloroquine (Aralen®); cyclophosphamide (Cytoxan®), methotrexate (Rheumatrex®), azathioprine (Imuran®), and anticoagulants such as heparin (Calcinparine® or Liquaemin®) and warfarin (Coumadin®).

[0263] In some embodiments, the present invention provides a method of treating Crohn's disease, ulcerative colitis, or inflammatory bowel disease, comprising administering to a patient in need thereof a compound of Formula I and one or more additional therapeutic agents selected from antidiarrheal agents such as mesalamine (Asacol®), sulfasalazine (Azulfidine®), diphenoxylate (Lomotil®), and loperamide (Imodium®), bile acid binding agents such as cholestyramine, laxatives such as alosetron (Lotronex®), lubiprostone (Amitiza®), milk of magnesia, polyethylene glycol (MiraLax®), Dulcolax®, Correctol®, and Senokot®, and anticholinergics or antispasmodics such as dicyclomine (Bentyl®), anti-TNF therapy, steroids, and antibiotics such as flagyl or ciprofloxacin.

[0264] In some embodiments, the present invention provides a method of treating asthma, comprising administering to a patient in need thereof a compound of Formula I and a beta-2 agonist such as Singulair®, albuterol (Ventolin® HFA, Proventil® HFA), levalbuterol (Xopenex®), metaproterenol (Arpent®), pirbuterol acetate (Maxair®), terbutaline sulfate (Brethaire®), salmeterol xinafoate (Serevent®), and formoterol (Foradil®), an anticholinergic agent such as ipratropium bromide (Atrovent®) and tiotropium (Spiriva®), prednisone, prednisolone, beclomethasone dipropionate (Becloven®), or a steroid agent such as benzodiazepine (Bordeaux®). and one or more additional therapeutic agents selected from inhaled corticosteroids such as fluticasone (Azmacort®), flumethasone (Asthmanex®), triamcinolone acetonide (Azmacort®), mometasone (Asthmanex®), budesonide (Pulmocort®), flunisolide (Aerobid®), Afviar®, Symbicort®, and Dulera®; methylxanthines such as cromolyn sodium (Intal®), theophylline (Theo-Dur®, Theolair®, Slo-bid®, Uniphyl®, Theo-24®) and aminophylline; and IgE antibodies such as omalizumab (Xolair®).

[0265] In some embodiments, the present invention provides a method of treating COPD, comprising administering to a patient in need thereof a compound of Formula I and a beta-2 agonist such as albuterol (Ventolin® HFA, Proventil® HFA), levalbuterol (Xopenex®), metaproterenol (Arpent®), pirbuterol acetate (Maxair®), terbutaline sulfate (Brethaire®), salmeterol xinafoate (Serevent®), and formoterol (Foradil®), an anticholinergic agent such as ipratropium bromide (Atrovent®) and tiotropium (Spiriva®), theophylline (Theo-Dur and one or more additional therapeutic agents selected from methylxanthines such as methylxanthines (e.g., Azolam, Theolair®, Slo-bid®, Uniphyl®, Theo-24®) and aminophylline; inhaled corticosteroids such as prednisone, prednisolone, beclomethasone dipropionate (Beclovent®, Qvar®, and Vanceril®), triamcinolone acetonide (Azmacort®), mometasone (Asthmanex®), budesonide (Pulmocort®), flunisolide (Aerobid®), Afviar®, Symbicort®, and Dulera®.

[0266] In another embodiment, the invention provides a method of treating a hematological malignancy, comprising administering to a patient in need thereof a compound of Formula I and one or more additional therapeutic agents selected from rituximab (Rituxan®), cyclophosphamide (Cytoxan®), doxorubicin (Hydrodaunorubicin®), vincristine (Oncovin®), prednisone, a hedgehog signaling inhibitor, a BTK inhibitor, a JAK / pan-JAK inhibitor, a PI3K inhibitor, a SYK inhibitor, and combinations thereof.

[0267] In another embodiment, the invention provides a method of treating a solid tumor, comprising administering to a patient in need thereof a compound of Formula I and one or more additional therapeutic agents selected from rituximab (Rituxan®), cyclophosphamide (Cytoxan®), doxorubicin (Hydrodaunorubicin®), vincristine (Oncovin®), prednisone, a hedgehog signaling inhibitor, a BTK inhibitor, a JAK / pan-JAK inhibitor, a PI3K inhibitor, a SYK inhibitor, and combinations thereof.

[0268] In another embodiment, the invention provides a method of treating a hematological malignancy, comprising administering to a patient in need thereof a compound of Formula I and a hedgehog (Hh) signaling pathway inhibitor. In some embodiments, the hematological malignancy is DLBCL (Ramirez et al., "Defining causative factors contributing in the activation of hedgehog signaling in diffuse large B-cell lymphoma," Leuk. Res. (2012), published online July 17, and incorporated herein by reference in its entirety).

[0269] In another embodiment, the invention provides a method of treating diffuse large B-cell lymphoma (DLBCL), comprising administering to a patient in need thereof a compound of Formula I and one or more additional therapeutic agents selected from rituximab (Rituxan®), cyclophosphamide (Cytoxan®), doxorubicin (Hydrodaunorubicin®), vincristine (Oncovin®), prednisone, a hedgehog signaling inhibitor, and combinations thereof.

[0270] In another embodiment, this invention provides a method of treating multiple myeloma, comprising administering to a patient in need thereof a compound of Formula I and one or more additional therapeutic agents selected from dexamethasone (Decadron®), a hedgehog signaling inhibitor, a BTK inhibitor, a JAK / pan-JAK inhibitor, a TYK2 inhibitor, a PI3K inhibitor, a SYK inhibitor, in combination with bortezomib (Velcade®), and lenalidomide (Revlimid®).

[0271] In another embodiment, the present invention provides a method of treating or lessening the severity of a disease, comprising administering to a patient in need thereof a compound of Formula I and a BTK inhibitor, wherein the disease is inflammatory bowel disease, arthritis, cutaneous lupus erythematosus, systemic lupus erythematosus (SLE), vasculitis, idiopathic thrombocytopenic purpura (ITP), rheumatoid arthritis, psoriatic arthritis, osteoarthritis, Still's disease, juvenile arthritis, diabetes, myasthenia gravis, Hashimoto's disease (chronic thyroiditis), Ord's thyroiditis (Ord's thyroiditis), or inflammatory bowel disease (inflammatory bowel disease). thyroiditis), Graves' disease, autoimmune thyroiditis, Sjogren's syndrome, multiple sclerosis, systemic sclerosis, Lyme neuroborreliosis, Guillain-Barré syndrome, acute disseminated encephalomyelitis, Addison's disease, opsoclonus-myoclonus syndrome, ankylosing spondylitis, antiphospholipid antibody syndrome, aplastic anemia, autoimmune hepatitis, autoimmune gastritis, pernicious anemia, celiac disease, Goodpasture's syndrome, idiopathic thrombocytopenic purpura, optic neuritis, Scleroderma, primary biliary cirrhosis, Reiter's syndrome, Takayasu's arteritis, temporal arteritis, warm autoimmune hemolytic anemia, Wegener's granulomatosis, psoriasis, alopecia universalis, Behçet's disease, chronic fatigue, dysautonomia, membranous glomerulonephritis, endometriosis, interstitial cystitis, pemphigus vulgaris, bullous pemphigoid, neuromyotonia, scleroderma, vulvodynia, hyperproliferative disorders, rejection of transplanted organs or tissues, acquired immune deficiency syndrome (AIDS, also known as HIV) , type 1 diabetes, graft-versus-host disease, transplants, blood transfusions, anaphylaxis, allergies (e.g., allergies to plant pollen, latex, drugs, foods, insect venom, animal hair, animal dander, dust mites, or cockroach umbrellas), type 1 hypersensitivity, allergic conjunctivitis, allergic rhinitis, and atopic dermatitis, asthma, appendicitis, atopic dermatitis, asthma, allergies, blepharitis, bronchiolitis, bronchitis, bursitis, cervicitis, cholangitis, cholecystitis, chronic graft rejection Diarrhea, colitis, conjunctivitis, Crohn's disease, cystitis, dacryoadenitis, dermatitis, dermatomyositis, encephalitis, endocarditis, endometritis, enteritis, gastroenteritis, epicondylitis, epididymitis, fasciitis, fibrosis, gastritis, gastroenteritis, Henoch-Schönlein purpura, hepatitis, hidradenitis suppurativa, immunoglobulin A nephropathy, interstitial lung disease, laryngitis, mastitis, meningitis, myelitis, myocarditis, myositis, nephritis, oophoritis, orchitis, osteitis, otitis, pancreatitis, parotitis, pericarditis, peritonitis, pharyngitis, pleuritis, phlebitis, pneumonitis, pneumonia,polymyositis, proctitis, prostatitis, pyelonephritis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis, tendonitis, tonsillitis, ulcerative colitis, uveitis, vaginitis, vasculitis, or vulvitis, B-cell proliferative disorders such as diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic lymphoma, chronic lymphocytic leukemia, acute lymphocytic leukemia, B-cell lymphocytic leukemia, lymphoplasmacytic lymphoma / Waldenstrom's macroglobulinemia, splenic marginal zone lymphoma, multiple myeloma (also known as plasma cell myeloma), non-Hodgkin's lymphoma, Hodgkin's lymphoma, plasmacytoma , extranodal marginal zone B-cell lymphoma, nodal marginal zone B-cell lymphoma, mantle cell lymphoma, mediastinal (thymic) large B-cell lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, Burkitt's lymphoma / leukemia, also lymphomatoid granulomatosis, breast cancer, prostate cancer, or mast cell cancer (e.g., mastocytoma, mast cell leukemia, mast cell sarcoma, systemic mastocytosis), bone cancer, colorectal cancer, pancreatic cancer, including but not limited to rheumatoid arthritis, seronegative spondyloarthropathy (including ankylosing spondylitis, psoriatic arthritis, and Reiter's disease), Behcet's disease, Schneider's disease, Glenn's syndrome, systemic sclerosis, osteoporosis, bone cancer, bone and joint diseases including bone metastases, thromboembolic disorders (e.g., myocardial infarction, angina pectoris, re-occlusion after angioplasty, restenosis after angioplasty, re-occlusion after aortocoronary artery bypass surgery, restenosis after aortocoronary artery bypass surgery, stroke, transient cerebral ischemia, peripheral arterial occlusive disease, pulmonary embolism, deep vein thrombosis), pelvic inflammatory disease, urethritis, sunburn, sinusitis, pneumonia, encephalitis, meningitis, myocarditis, nephritis, osteomyelitis, myositis, hepatitis, gastritis, enteritis, dermatitis, gingivitis, appendicitis, pancreatitis, cholecystitis, agammaglobulinemia, psoriasis, allergies, Crohn's disease, irritable bowel syndrome, ulcerative colitis, Sjogren's syndrome, tissue graft rejection, hyperacute rejection of transplanted organs, asthma, allergic rhinitis, chronic obstructive pulmonary disease (COPD), autoimmune polyendocrine disease (also known as autoimmune polyendocrine syndrome), autoimmune alopecia, pernicious anemia, glomerulonephritis, dermatomyositis, multiple sclerosis, scleroderma, vasculitis, autoimmune hemolytic and thrombocytopenic states, Goodpasture's syndrome, atherosclerosis, Addison's disease, Parkinson's disease, Alzheimer's disease, diabetes, septic shock, cutaneous lupus erythematosus,Selected from systemic lupus erythematosus (SLE), rheumatoid arthritis, psoriatic arthritis, juvenile arthritis, osteoarthritis, chronic idiopathic thrombocytopenic purpura, Waldenstrom's macroglobulinemia, myasthenia gravis, Hashimoto's disease (chronic thyroiditis), atopic dermatitis, degenerative joint disease, vitiligo, autoimmune hypopituitarism, Guillain-Barré syndrome, Behcet's disease, scleroderma, mycosis fungoides, acute inflammatory reactions (such as acute respiratory distress syndrome and ischemia / reperfusion injury), and Graves' disease.

[0272] In another embodiment, the invention provides a method of treating or lessening the severity of a disease, comprising administering to a patient in need thereof a compound of Formula I and a PI3K inhibitor, wherein the disease is selected from cancer, a neurodegenerative disorder, an angiogenic disorder, a viral disease, an autoimmune disease, an inflammatory disorder, a hormone-related disease, a condition associated with organ transplantation, an immunodeficiency disorder, a destructive bone disorder, a proliferative disorder, an infectious disease, a condition associated with cell death, thrombin-induced platelet aggregation, chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), liver disease, a pathological immune condition involving T-cell activation, a cardiovascular disorder, and a CNS disorder.

[0273] In another embodiment, the invention provides a method of treating or lessening the severity of a disease, comprising administering to a patient in need thereof a compound of Formula I and a PI3K inhibitor, wherein the disease is a benign or malignant tumor, carcinoma or solid tumor of the brain, kidney (e.g., renal cell carcinoma (RCC)), liver, adrenal gland, bladder, breast, stomach, gastric tumor, ovary, colon, rectum, prostate, pancreas, lung, vagina, endometrium, cervix, testis, genitourinary tract, esophagus, larynx, skin, bone, or thyroid, sarcoma, glioblastoma, neuroblastoma, multiple myeloma, or gastrointestinal cancer, particularly colon cancer or colorectal adenoma, or head and neck cancer. Diseases including tumors, epidermal hyperproliferation, psoriasis, benign prostatic hyperplasia, neoplasia, neoplasia of epithelial character, adenoma, adenocarcinoma, keratoacanthoma, epidermoid carcinoma, large cell carcinoma, non-small cell lung cancer, lymphoma (including, for example, non-Hodgkin's lymphoma (NHL), Hodgkin's lymphoma (Hodgkin's lymphoma, or Hodgkin's disease)), breast cancer, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, or leukemia, Cowden syndrome, Lhermitte-Dacros disease, and Banayan-Zonana syndrome, or diseases in which the PI3K / PKB pathway is abnormally activated, intrinsic (non-allergic) asthma and extrinsic (allergic) asthma asthma of any type or genesis, including acute, arachidic, catarrhal, croupy, chronic, or non-allergic asthma, both mild asthma, moderate asthma, severe asthma, bronchial asthma, exercise-induced asthma, occupational asthma, and asthma induced after bacterial infection, acute lung injury (ALI), adult / acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease, airway or lung disease (COPD, COAD, or COLD), including chronic bronchitis or related dyspnea, emphysema, and exacerbations of airway hyperresponsiveness as a result of other medications, especially other inhaled medications; bronchitis of any kind or genesis, including but not limited to tuberculoid bronchitis; pneumoconiosis of any kind or genesis (an inflammatory, generally occupational, lung disease, whether acute or chronic, often complicated by airway obstruction and caused by repeated dust inhalation) (including, for example, aluminum lung disease, anthracosis, asbestosis, stone disease, ptilosis, siderosis, silicosis, tobacco disease, and byssinosis); Löffler's syndrome; eosinophilic pneumonia; parasitic (especially metazoan) infestations (including tropical eosinophilia); bronchopulmonary aspergillosis;Polyarteritis nodosa (including Churg-Strauss syndrome), eosinophilic granuloma, and drug reactions, psoriasis, contact dermatitis, atopic dermatitis, alopecia areata, erythema multiforme, dermatitis herpetiformis, scleroderma, vitiligo, hypersensitivity vasculitis, urticaria, bullous pemphigoid, lupus erythematosus, pemphigus, epidermolysis bullosa acquisita, eosinophil-related disorders affecting the airways caused by conjunctivitis, keratoconjunctivitis sicca, and vernal keratoconjunctivitis, diseases affecting the nose, including allergic rhinitis, and inflammatory diseases in which an autoimmune response is involved or has an autoimmune component or etiology (autoimmune blood disorders (e.g., hemolytic anemia, aplastic anemia, pure red cell aplasia, and idiopathic thrombocytopenia) keratoconjunctivitis, cutaneous lupus erythematosus, systemic lupus erythematosus, rheumatoid arthritis, polychondritis, scleroderma, Wegener's granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, Stevens-Johnson syndrome, idiopathic sprue, autoimmune inflammatory bowel disease (e.g., ulcerative colitis and Crohn's disease), endocrine ophthalmopathy, Graves' disease, sarcoidosis, alveolitis, chronic hypersensitivity pneumonitis, multiple sclerosis, primary biliary cirrhosis, uveitis (pre- and post-operative), keratoconjunctivitis sicca and vernal keratoconjunctivitis, interstitial pulmonary fibrosis, psoriatic arthritis, and glomerulonephritis (with or without nephrotic syndrome (e.g., idiopathic nephrotic syndrome or minimal change nephropathy)). The therapeutic agent is selected from neurodegenerative diseases caused by traumatic injury, glutamate neurotoxicity, and hypoxia, including cerebrovascular disease (including cerebrovascular accidents), restenosis, cardiac hypertrophy, atherosclerosis, myocardial infarction, ischemic stroke, and congestive heart failure, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, and cerebral ischemia, and neurodegenerative diseases caused by traumatic injury, glutamate neurotoxicity, and hypoxia.

[0274] In some embodiments, the invention provides a method of treating or reducing the severity of a disease, comprising administering to a patient in need thereof a compound of Formula I and a Bcl-2 inhibitor, wherein the disease is an inflammatory disorder, an autoimmune disorder, a proliferative disorder, an endocrine disorder, a neurological disorder, or a transplant-related disorder. In some embodiments, the disorder is a proliferative disorder, lupus, or lupus nephritis. In some embodiments, the proliferative disorder is chronic lymphocytic leukemia, diffuse large B-cell lymphoma, Hodgkin's lymphoma, small cell lung cancer, non-small cell lung cancer, myelodysplastic syndrome, lymphoma, hematological neoplasm, or solid tumor.

[0275] In some embodiments, the disease is an autoimmune disorder, an inflammatory disorder, a proliferative disorder, an endocrine disorder, a neurological disorder, or a transplant-related disorder. In some embodiments, the JH2 binding compound is a compound of Formula I. Other suitable JH2 domain binding compounds include those described in WO2014 / 074660A1, WO2014 / 074661A1, and WO2015 / 089143A1, each of which is incorporated by reference in its entirety. Suitable JH1 domain binding compounds include those described in WO2015 / 131080A1, each of which is incorporated by reference in its entirety.

[0276] The compounds and compositions according to the methods of the present invention may be administered in any amount and using any route of administration effective to treat or reduce the severity of an autoimmune, inflammatory, proliferative, endocrine, neurological, or transplant-related disorder. The exact amount required will vary from subject to subject, depending on the subject's species, age, and general condition, the severity of the infection, the particular drug, its mode of administration, and the like. The compounds of the present invention are preferably formulated in unit dosage form for ease of administration and uniformity of dosage. As used herein, the term "unit dosage form" refers to a physically discrete pharmaceutical unit appropriate for the patient being treated. However, it will be understood that the total daily usage of the compounds and compositions of the present invention will be determined by the attending physician within the scope of sound medical judgment. The specific effective dosage level for any particular patient or organism will depend on a variety of factors, including the disorder and severity of the disorder being treated, the activity of the particular compound employed, the particular composition employed, the patient's age, weight, general health, sex, and diet, the time, route of administration, and excretion rate of the particular compound employed, the duration of treatment, drugs used in combination with or concurrently with the particular compound employed, and similar factors well known in the medical arts. The term "patient", as used herein, means an animal, preferably a mammal, and most preferably a human.

[0277] The pharmaceutically acceptable compositions of the present invention can be administered to humans and other animals orally, rectally, parenterally, intravesically, intravaginally, intraperitoneally, topically (by powder, ointment, or drops), buccally, as an oral or nasal spray, or the like, depending on the severity of the infection being treated. In certain embodiments, the compounds of the present invention may be administered orally or parenterally, one or more times daily, at dosage levels of about 0.01 mg / kg to about 50 mg / kg, and preferably about 1 mg / kg to about 25 mg / kg, of the subject's body weight per day to achieve the desired therapeutic effect.

[0278] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs.In addition to the active compound, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers, emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and / or sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof.In addition to inert diluents, oral compositions may also contain adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and fragrances.

[0279] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions, can be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations can also be sterile injectable solutions, suspensions, or emulsions in non-toxic parenterally acceptable diluents or solvents, for example, as solutions in 1,3-butanediol. Among the acceptable vehicles and solvents that can be used are water, Ringer's solution (USP), and isotonic sodium chloride solution. Additionally, sterile fixed oils are conventionally used as solvents or suspending media. For this purpose, any non-irritating fixed oil can be used, including synthetic monoglycerides or diglycerides. Additionally, fatty acids such as oleic acid are used in the preparation of injectables.

[0280] Injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium before use.

[0281] In order to prolong the effect of a compound of the present invention, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This can be accomplished by using a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound then depends on its rate of dissolution, which may depend on crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form can be accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of compound to polymer and the nature of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations can also be prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.

[0282] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing a compound of the invention with a suitable non-irritating excipient or carrier such as cocoa butter, polyethylene glycol, or a suppository wax which is solid at ambient temperature but liquid at body temperature and therefore melts in the rectum or vaginal cavity and releases the active compound.

[0283] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and / or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retardants such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) humectants such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0284] Solid compositions of a similar type may also be used as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like. Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They may optionally contain opacifying agents and can be of a composition that releases the active ingredient only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be used as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like.

[0285] The active compound may also be in microencapsulated form with one or more excipients, as described above. Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings, release-controlling coatings, and other coatings well known in the pharmaceutical formulation art. In such solid dosage forms, the active compound may be mixed with at least one inert diluent, such as sucrose, lactose, or starch. Such dosage forms may also contain, as is common practice, additional substances other than inert diluents, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage forms may also contain buffering agents. They may optionally contain opacifying agents, and may be of a composition that releases the active ingredient only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

[0286] Dosage forms for topical or transdermal administration of the compounds of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. The active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers, if required. Ophthalmic formulations, ear drops, and eye drops are also contemplated within the scope of the present invention. Furthermore, the present invention contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of the compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0287] According to one embodiment, the present invention relates to a method of inhibiting protein kinase activity in a biological sample, comprising the step of contacting said biological sample with a compound of the present invention, or a composition comprising said compound.

[0288] According to another embodiment, the present invention relates to a method for inhibiting the activity of HPK1 or a mutant thereof in a biological sample, comprising contacting the biological sample with a compound of the present invention, or a composition comprising the compound. In certain embodiments, the present invention relates to a method for irreversibly inhibiting HPK1 or a mutant thereof, activity in a biological sample, comprising contacting the biological sample with a compound of the present invention, or a composition comprising the compound.

[0289] As used herein, the term "biological sample" includes, but is not limited to, cell cultures or extracts thereof, biopsies obtained from mammals or extracts thereof, as well as blood, saliva, urine, feces, semen, tears, or other bodily fluids or extracts thereof.

[0290] Inhibition of HPK1 (or variants thereof) activity in a biological sample is useful for a variety of purposes known to those of skill in the art, including, but not limited to, blood transfusion, organ transplantation, biological specimen storage, and biological assays.

[0291] Another embodiment of the invention relates to a method of inhibiting protein kinase activity in a patient, comprising administering to said patient a compound of the invention or a composition comprising said compound.

[0292] According to another embodiment, the present invention relates to a method of inhibiting the activity of HPK1 or a mutant thereof in a patient, comprising administering to the patient a compound of the present invention or a composition comprising the compound. According to certain embodiments, the present invention relates to a method of reversibly or irreversibly inhibiting HPK1 or one or more mutants thereof in a patient, comprising administering to the patient a compound of the present invention or a composition comprising the compound. In other embodiments, the present invention provides a method for treating a disorder mediated by HPK1 or a mutant thereof in a patient in need thereof, comprising administering to the patient a compound according to the present invention or a pharmaceutically acceptable composition thereof. Such disorders are described in detail herein.

[0293] Depending on the particular condition, or disease, being treated, additional therapeutic agents, which are normally administered to treat that condition, may also be present in the compositions of this invention. As used herein, additional therapeutic agents that are normally administered to treat a particular disease or condition are known as "appropriate for the disease, or condition, being treated."

[0294] The compounds of the invention may also be used to advantage in combination with other therapeutic compounds, hi some embodiments, the other therapeutic compounds are antiproliferative compounds. Such antiproliferative compounds include aromatase inhibitors; antiestrogens; topoisomerase I inhibitors; topoisomerase II inhibitors; microtubule active compounds; alkylating compounds; histone deacetylase inhibitors; compounds that induce cell differentiation processes; cyclooxygenase inhibitors; MMP inhibitors; mTOR inhibitors; antineoplastic antimetabolites; platinum compounds; compounds that target / reduce protein or lipid kinase activity and further antiangiogenic compounds; compounds that target, reduce or inhibit the activity of protein or lipid phosphatases; gonadorelin agonists; antiandrogens; methionine aminopeptidase inhibitors; matrix metalloproteinase inhibitors; bisphosphonates; biological response modifiers; antiproliferative antibodies; heparanase inhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasome inhibitors; compounds used in the treatment of hematological malignancies; compounds that target, reduce or inhibit the activity of Flt-3; Conforma Hsp90 inhibitors such as 17-AAG (17-allylaminogeldanamycin, NSC330507), 17-DMAG (17-dimethylaminoethylamino-17-demethoxy-geldanamycin, NSC707545), IPI-504, CNF1010, CNF2024, CNF1010 from Therapeutics; kinesin spindle protein inhibitors such as temozolomide (Temodal®); SB715992 or SB743921 from GlaxoSmithKline, or pentamidine / chlorpromazine from CombinatoRx; MEK inhibitors such as ARRY142886 from Array BioPharma, AZD6244 from AstraZeneca, PD181461 from Pfizer, and leucovorin. The term "aromatase inhibitor", as used herein, relates to compounds which inhibit estrogen production, eg the conversion of the substrates androstenedione and testosterone to estrone and estradiol, respectively.The term includes, but is not limited to, steroids, particularly atamestane, exemestane, and formestane, and nonsteroids, particularly aminoglutethimide, rogletimide, pyridoglutethimide, trilostane, testolactone, ketoconazole, vorozole, fadrozole, anastrozole, and letrozole. Exemestane is sold under the trade name Aromasin™. Formestane is sold under the trade name Lentaron™. Fadrozole is sold under the trade name Afema™. Anastrozole is sold under the trade name Arimidex™. Letrozole is sold under the trade name Femara™ or Femar™. Aminoglutethimide is sold under the trade name Orimeten™. Combinations of the present invention containing a chemotherapeutic agent that is an aromatase inhibitor are particularly useful in the treatment of hormone receptor-positive tumors, such as breast tumors.

[0295] As used herein, the term "anti-estrogen" refers to a compound that antagonizes the effects of estrogen at the estrogen receptor level. This term includes, but is not limited to, tamoxifen, fulvestrant, raloxifene, and raloxifene hydrochloride. Tamoxifen is sold under the trade name Nolvadex™. Raloxifene hydrochloride is sold under the trade name Evista™. Fulvestrant can be administered under the trade name Faslodex™. The combination of the present invention, which includes a chemotherapeutic agent that is an anti-estrogen, is particularly useful for treating estrogen receptor-positive tumors, such as breast tumors.

[0296] As used herein, the term "antiandrogen" refers to any substance capable of inhibiting the biological effects of androgen hormones, including, but not limited to, bicalutamide (Casodex™). As used herein, the term "gonadorelin agonist" includes, but is not limited to, abarelix, goserelin, and goserelin acetate. Goserelin can be administered under the trade name Zoladex™.

[0297] As used herein, the term "topoisomerase I inhibitor" includes, but is not limited to, topotecan, gimatecan, irinotecan, camptothecin and its analogs, 9-nitrocamptothecin, and the polymeric camptothecin conjugate PNU-166148. Irinotecan can be administered in the form as it is marketed, for example, under the trademark Camptosar™. Topotecan is sold under the trade name Hycamptin™.

[0298] As used herein, the term "topoisomerase II inhibitors" includes, but is not limited to, doxorubicin (including liposomal formulations such as Caelyx™), anthracyclines such as daunorubicin, epirubicin, idarubicin, and nemorubicin, the anthraponones mitoxantrone and losoxantrone, and the podophyllotoxins etoposide and teniposide. Etoposide is sold under the trade name Etopophos™. Teniposide is sold under the trade name VM 26-Bristol. Doxorubicin is sold under the trade name Acriblastin™ or Adriamycin™. Epirubicin is sold under the trade name Farmorubicin™. Idarubicin is sold under the trade name Zavedos™. Mitoxantrone is sold under the trade name Novantron.

[0299] The term "microtubule active agent" refers to microtubule stabilizing, microtubule destabilizing compounds and microtubule polymerization inhibitors, including but not limited to taxanes such as paclitaxel and docetaxel; vinca alkaloids such as vinblastine or vinblastine sulfate, vincristine or vincristine sulfate, and vinorelbine; discodermolide; colchicine and epothilones and their derivatives. Paclitaxel is sold under the trade name Taxol™. Docetaxel is sold under the trade name Taxotere™. Vinblastine sulfate is sold under the trade name Vinblastin RP™. Vincristine sulfate is sold under the trade name Farmistin™.

[0300] As used herein, the term "alkylating agent" includes, but is not limited to, cyclophosphamide, ifosfamide, melphalan, or nitrosoureas (BCNU or Gliadel). Cyclophosphamide is sold under the trade name Cyclostin™. Ifosfamide is sold under the trade name Holoxan™.

[0301] The term "histone deacetylase inhibitors" or "HDAC inhibitors" relates to compounds which inhibit histone deacetylase and which possess antiproliferative activity, including, but not limited to, suberoylanilide hydroxamic acid (SAHA).

[0302] The term "antineoplastic antimetabolite" includes, but is not limited to, 5-fluorouracil or 5-FU, DNA demethylating compounds such as capecitabine, gemcitabine, 5-azacytidine, and decitabine, methotrexate and edatrexate, and folate antagonists such as pemetrexed. Capecitabine is sold under the trade name Xeloda™. Gemcitabine is sold under the trade name Gemzar™.

[0303] As used herein, the term "platinum compounds" includes, but is not limited to, carboplatin, cis-platin, cisplatinum, and oxaliplatin. Carboplatin can be administered in the form as it is marketed, e.g., under the trademark Carboplat™. Oxaliplatin can be administered in the form as it is marketed, e.g., under the trademark Eloxatin™.

[0304] The term "compounds which target / reduce protein or lipid kinase activity, or protein or lipid phosphatase activity; or further anti-angiogenic compounds" refers to a) compounds which target, reduce or inhibit the activity of PDGFR, in particular compounds which inhibit PDGF receptors, e.g., N-phenyl-2-pyrimidine-amine derivatives, e.g., imatinib, SU101, SU6668, and GFB-111, which target, reduce or inhibit the activity of platelet-derived growth factor receptors (PDGFR), b) compounds which target, reduce or inhibit the activity of PDGFR, in particular compounds which inhibit PDGF receptors, e.g., N-phenyl-2-pyrimidine-amine derivatives, e.g., imatinib, SU101, SU6668, and GFB-111, c) compounds which target, reduce or inhibit the activity of PDGFR, such as fibroblast growth factor receptors (GFAP), c) compounds that target, reduce or inhibit the activity of IGF-IR, in particular compounds that target, reduce or inhibit the activity of insulin-like growth factor receptor 1 (IGF-IR), such as compounds that inhibit the kinase activity of the IGF-I receptor or antibodies that target the extracellular domain of the IGF-I receptor or its growth factors; d) compounds that target, reduce or inhibit the activity of the Trk receptor tyrosine kinase family or ephrin B4 inhibitors, e) compounds which target, decrease or inhibit the activity of the AxI receptor tyrosine kinase family, f) compounds which target, decrease or inhibit the activity of the Ret receptor tyrosine kinase, g) compounds which target, decrease or inhibit the activity of the Kit / SCFR receptor tyrosine kinase, e.g., imatinib, h) compounds which target, decrease or inhibit the activity of the C-kit receptor tyrosine kinase, which is part of the PDGFR family, e.g., compounds which target the activity of the c-Kit receptor tyrosine kinase family. i) compounds that target, reduce or inhibit the activity of c-Abl family members, their gene fusion products (e.g. BCR-Abl kinase) and mutants, for example compounds that target, reduce or inhibit the activity of c-Abl family members and their gene fusion products, for example N-phenyl-2-pyrimidine-amine derivatives, for example imatinib or nilotinib (AMN107); PD180970;AG957; NSC 680410; PD173955 from ParkeDavis; or dasatinib (BMS-354825), j) compounds that target, decrease, or inhibit the activity of members of the protein kinase C (PKC) and Raf family of serine / threonine kinases, MEK, SRC, JAK / pan-JAK, FAK, PDK1, PKB / Akt, Ras / MAPK, PI3K, SYK, BTK, and TEC families, and / or members of the cyclin-dependent kinase family (CDK) (including staurosporine derivatives such as midostaurin) (further examples of compounds include UCN-01, safingol, BAY 43-9006, bryostatin 1, perifosine; irmofosine; RO 318220 and RO 320432; GO 6976; lsis 3521; LY333531 / LY379196; isoquinoline compounds; FTIs; PD184352 or QAN697 (P13K inhibitors) or AT7519 (CDK inhibitors)), k) compounds which target, decrease or inhibit the activity of protein tyrosine kinase inhibitors, for example protein tyrosine kinase inhibitors (imatinib mesylate (Gleevec™) or tyrphostins, for example Tyrphostin A23 / RG-50810; AG 99; Tyrphostin AG 213; Tyrphostin AG 1748; Tyrphostin AG 490; Tyrphostin B44; Tyrphostin B44(+) enantiomer; Tyrphostin AG 555; AG 494; Tyrphostin AG 556, AG957, and adaphostine (4-{[(2,5-dihydroxyphenyl)methyl]amino}-benzoic acid adamantyl ester;l) compounds which target, decrease or inhibit the activity of epidermal growth factor family receptor tyrosine kinases (EGFR1, ErbB2, ErbB3, ErbB4 as homodimers or heterodimers) and variants thereof, for example, compounds which inhibit members of the EGF receptor tyrosine kinase family, such as, in particular, EGF receptor, ErbB2, ErbB3, and ErbB4, or EGF or EGF-related ligands, CP 358774, ZD 1839, ZM 105180;compounds that target, decrease, or inhibit the activity of the epidermal growth factor receptor family, such as trastuzumab (Herceptin™), cetuximab (Erbitux™), Iressa, Tarceva, OSI-774, Cl-1033, EKB-569, GW-2016, E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3, or E7.6.3, and compounds, proteins, or antibodies that bind to 7H-pyrrolo-[2,3-d]pyrimidine derivatives; m) compounds that target, decrease, or inhibit the activity of the epidermal growth factor receptor family, such as trastuzumab (Herceptin™), cetuximab (Erbitux™), Iressa, Tarceva, OSI-774, Cl-1033, EKB-569, GW-2016, E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3, or E7.6.3, and 7H-pyrrolo-[2,3-d]pyrimidine derivatives; n) one of the following compounds: a compound that targets, decreases, or inhibits the activity of the c-Met receptor, in particular a compound that inhibits the kinase activity of the c-Met receptor, or an antibody that targets the extracellular domain of c-Met or binds to HGF; n) one of the following compounds: PRT-062070, SB-1578, baricitinib, pacritinib, momelotinib, VX-509, AZD-1480, TG-101348, tofacitinib, and ruxolitinib; o) compounds that target, decrease, or inhibit the kinase activity of the above JAK family members (JAK1 / JAK2 / JAK3 / TYK2 and / or pan-JAK); o) compounds that target, decrease, or inhibit the kinase activity of PI3 kinase (PI3K) kinases, including but not limited to ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, buparlisib, pictorelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765, and idelalisib; q) protein tyrosine kinase and / or serine and / or threonine kinase inhibitors or lipid kinase inhibitors, such as compounds that target, decrease or inhibit the activity of phosphodiesterases, and q) compounds that target, decrease or inhibit the signaling effects of the Hedgehog (Hh) or Smoothened receptor (SMO) pathways, including but not limited to cyclopamine, vismodegib, itraconazole, erismodegib, and IPI-926 (salidegib);

[0305] As used herein, the term "PI3K inhibitor" includes, but is not limited to, compounds that have inhibitory activity against one or more enzymes in the phosphatidylinositol-3-kinase family, including, but not limited to, PI3K α, PI3K γ, PI3K δ, PI3K β, PI3K-C2α, PI3K-C2β, PI3K-C2γ, Vps34, p110-α, p110-β, p110-γ, p110-δ, p85-α, p85-β, p55-γ, p150, p101, and p87. Examples of PI3K inhibitors useful in the present invention include ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, buparlisib, pictorelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765, and idelalisib.

[0306] As used herein, the term "BTK inhibitor" includes, but is not limited to, compounds that have inhibitory activity against Bruton's tyrosine kinase (BTK), including, but not limited to, AVL-292 and ibrutinib.

[0307] As used herein, the term "SYK inhibitor" includes, but is not limited to, compounds that have inhibitory activity against spleen tyrosine kinase (SYK), including, but not limited to, PRT-062070, R-343, R-333, Excellair, PRT-062607, and fostamatinib.

[0308] As used herein, the term "Bcl-2 inhibitor" includes but is not limited to ABT-199, ABT-731, ABT-737, apogossypol, the pan-Bcl-2 inhibitors of Ascenta, curcumin (and analogs thereof), dual Bcl-2 / Bcl-xL inhibitors (Infinity Pharmaceuticals / Novartis Pharmaceuticals), Genasense (G3139), HA14-1 (and analogs thereof, see WO2008 / 118802), navitoclax (and analogs thereof, see US7390799), NH-1 (Shenayng Pharmaceutical University), obatoclax (and analogs thereof, see WO2004 / 106328), S-001 (Gloria Pharmaceuticals), TW series compounds (Univ. of Examples of compounds that have inhibitory activity against B-cell lymphoma 2 protein (Bcl-2), including but not limited to, cefotaxime (C271), ...

[0309] Further examples of BTK inhibitory compounds and conditions treatable by such compounds in combination with the compounds of the present invention can be found in WO2008 / 039218 and WO2011 / 090760, which are incorporated herein by reference in their entireties.

[0310] Further examples of SYK inhibitory compounds and conditions treatable by such compounds in combination with the compounds of the present invention can be found in WO2003 / 063794, WO2005 / 007623, and WO2006 / 078846, which are incorporated herein by reference in their entireties.

[0311] Further examples of PI3K inhibitory compounds and conditions treatable by such compounds in combination with the compounds of the present invention can be found in WO2004 / 019973, WO2004 / 089925, WO2007 / 016176, US8138347, WO2002 / 088112, WO2007 / 084786, WO2007 / 129161, WO2006 / 122806, WO2005 / 113554, and WO2007 / 044729, which are incorporated herein by reference in their entireties.

[0312] Further examples of JAK inhibitory compounds and conditions treatable by such compounds in combination with the compounds of the present invention can be found in WO2009 / 114512, WO2008 / 109943, WO2007 / 053452, WO2000 / 142246, and WO2007 / 070514, which are incorporated herein by reference in their entireties.

[0313] Additional anti-angiogenic compounds include, for example, compounds that have another mechanism for their activity not related to protein or lipid kinase inhibition, such as thalidomide (Thalomid™) and TNP-470.

[0314] Examples of proteasome inhibitors useful in combination with the compounds of the invention include, but are not limited to, bortezomib, disulfiram, epigallocatechin-3-gallate (EGCG), salinosporamide A, carfilzomib, ONX-0912, CEP-18770, and MLN9708.

[0315] Compounds which target, decrease or inhibit the activity of a protein or lipid phosphatase are eg inhibitors of phosphatase 1, phosphatase 2A or CDC25, such as okadaic acid or a derivative thereof.

[0316] Compounds that induce cell differentiation processes include, but are not limited to, retinoic acid, α-, γ-, or δ-tocopherol, or α-, γ-, or δ-tocotrienol.

[0317] As used herein, the term cyclooxygenase inhibitors includes, but is not limited to, Cox-2 inhibitors, 5-alkyl substituted 2-arylaminophenylacetic acids and derivatives, such as celecoxib (Celebrex™), rofecoxib (Vioxx™), etoricoxib, valdecoxib, or 5-alkyl-2-arylaminophenylacetic acids, such as 5-methyl-2-(2'-chloro-6'-fluoroanilino)phenylacetic acid, lumiracoxib.

[0318] As used herein, the term "bisphosphonate" includes, but is not limited to, etridonic acid, clodronic acid, tiludronic acid, pamidronic acid, alendronic acid, ibandronic acid, risedronic acid, and zoledronic acid. Etridronic acid is sold under the trade name Didronel™. Clodronic acid is sold under the trade name Bonefos™. Tiludronic acid is sold under the trade name Skelid™. Pamidronic acid is sold under the trade name Aredia™. Alendronic acid is sold under the trade name Fosamax™. Ibandronic acid is sold under the trade name Bondranat™. Risedronic acid is sold under the trade name Actonel™. Zoledronic acid is sold under the trade name Zometa™. The term "mTOR inhibitors" relates to compounds which inhibit the mammalian target of rapamycin (mTOR) and which have antiproliferative activity, such as sirolimus (Rapamune®), everolimus (Certican™), CCI-779 and ABT578.

[0319] As used herein, the term "heparanase inhibitor" refers to a compound that targets, decreases, or inhibits heparin sulfate degradation. The term includes, but is not limited to, PI-88. As used herein, the term "biological response modifier" refers to a lymphokine or interferon.

[0320] As used herein, the term "inhibitors of Ras oncogenic isoforms," ​​such as H-Ras, K-Ras, or N-Ras, refers to compounds that target, reduce, or inhibit the oncogenic activity of Ras, e.g., "farnesyltransferase inhibitors," such as L-744832, DK8G557, or R115777 (Zarnestra™). As used herein, the term "telomerase inhibitors" refers to compounds that target, reduce, or inhibit the activity of telomerase. Compounds that target, reduce, or inhibit the activity of telomerase are, in particular, compounds that inhibit the telomerase receptor, such as telomestatin.

[0321] As used herein, the term "methionine aminopeptidase inhibitor" refers to a compound that targets, decreases or inhibits the activity of methionine aminopeptidase. Compounds that target, decrease or inhibit the activity of methionine aminopeptidase include, but are not limited to, bengamide or a derivative thereof.

[0322] As used herein, the term "proteasome inhibitor" refers to a compound that targets, decreases, or inhibits the activity of the proteasome. Compounds that target, decrease, or inhibit the activity of the proteasome include, but are not limited to, bortezomib (Velcade™) and MLN 341.

[0323] As used herein, the term "matrix metalloproteinase inhibitors" or ("MMP" inhibitors) includes, but is not limited to, collagen peptidomimetic and non-peptidomimetic inhibitors, tetracycline derivatives, such as the hydroxamate peptidomimetic inhibitor batimastat and its orally bioavailable analogs marimastat (BB-2516), prinomastat (AG3340), metastat (NSC 683551), BMS-279251, BAY 12-9566, TAA211, MMI270B, or AAJ996.

[0324] As used herein, the term "compounds used in the treatment of hematological malignancies" includes, but is not limited to, FMS-like tyrosine kinase inhibitors, which are compounds that target, decrease, or inhibit the activity of FMS-like tyrosine kinase receptor (Flt-3R); interferon, 1-β-D-arabinofuranosylcytosine (ara-c), and bisulfan; ALK inhibitors, which are compounds that target, decrease, or inhibit anaplastic lymphoma kinase; and Bcl-2 inhibitors.

[0325] Compounds that target, decrease or inhibit the activity of the FLT-3R-like tyrosine kinase receptor are, in particular, compounds, proteins or antibodies that inhibit members of the Flt-3R receptor kinase family, such as PKC412, midostaurin, staurosporine derivatives, SU11248, and MLN518.

[0326] As used herein, the term "HSP90 inhibitor" includes, but is not limited to, compounds that target, reduce, or inhibit the intrinsic ATPase activity of HSP90, such as degrading, targeting, reducing, or inhibiting HSP90 client proteins via the ubiquitin proteosome pathway. Compounds that target, reduce, or inhibit the intrinsic ATPase activity of HSP90 are, in particular, compounds, proteins, or antibodies that inhibit the ATPase activity of HSP90, such as 17-allylamino, 17-demethoxygeldanamycin (17AAG), geldanamycin derivatives; other geldanamycin-related compounds; radicicol, and HDAC inhibitors.

[0327] As used herein, the term "antiproliferative antibody" includes, but is not limited to, trastuzumab (Herceptin™), trastuzumab-DM1, Erbitux, bevacizumab (Avastin™), rituximab (Rituxan®), PRO64553 (anti-CD40), and 2C4 antibodies. Antibody refers to intact monoclonal antibodies, polyclonal antibodies, multispecific antibodies formed from at least two intact antibodies, and antibody fragments, so long as they exhibit the desired biological activity.

[0328] For the treatment of acute myeloid leukemia (AML), the compounds of the present invention can be used in combination with standard leukemia therapy, particularly in combination with therapy used to treat AML. In particular, the compounds of the present invention can be administered in combination with farnesyltransferase inhibitors and / or other drugs useful for the treatment of AML, such as daunorubicin, adriamycin, Ara-C, VP-16, teniposide, mitoxantrone, idarubicin, carboplatin, and PKC412. In some embodiments, the present invention provides a method for treating AML associated with ITD and / or D835Y mutation, comprising administering a compound of the present invention together with one or more FLT3 inhibitors. In some embodiments, the FLT3 inhibitor is selected from quizartinib (AC220), a staurosporine derivative (e.g., midostaurin or lestaurtinib), sorafenib, tandutinib, LY-2401401, LS-104, EB-10, famitinib, NOV-110302, NMS-P948, AST-487, G-749, SB-1317, S-209, SC-110219, AKN-028, fedratinib, tozasertib, and sunitinib. In some embodiments, the FLT3 inhibitor is selected from quizartinib, midostaurin, lestaurin, sorafenib, and sunitinib.

[0329] Other anti-leukemia compounds include, for example, the diamine 2-deoxycytidine ’These include Ara-C, a pyrimidine analogue that is an alpha-hydroxyribose (arabinoside) derivative. Also included are purine analogues of hypoxanthine, 6-mercaptopurine (6-MP) and fludarabine phosphate. Compounds that target, decrease or inhibit the activity of histone deacetylase (HDAC) inhibitors, such as sodium butyrate and suberoylanilide hydroxamic acid (SAHA), inhibit the activity of enzymes known as histone deacetylases. Specific HDAC inhibitors include MS275, SAHA, FK228 (formerly FR901228), trichostatin A, and compounds disclosed in US Pat. No. 6,552,065, including but not limited to N-hydroxy-3-[4-[[[2-(2-methyl-1H-indol-3-yl)-ethyl]-amino]methyl]phenyl]-2E-2-propenamide, or a pharmaceutically acceptable salt thereof, and N-hydroxy-3-[4-[(2-hydroxy){2-(1H-indol-3-yl)ethyl]-amino]methyl]phenyl]-2E-2-propenamide, or a pharmaceutically acceptable salt thereof, particularly the lactate salt. As used herein, somatostatin receptor antagonists refer to compounds that target, treat, or inhibit somatostatin receptors and SOM230, such as octreotide. Tumor cell damaging approaches refer to approaches such as ionizing radiation. The term "ionizing radiation" referred to above and below means ionizing radiation that occurs either as electromagnetic rays (such as X-rays and gamma rays) or particles (such as alpha and beta particles). Ionizing radiation is provided in, but is not limited to, radiation therapy and is known in the art. Hellman, Principles of Radiation Therapy, Cancer, Principles and Practice of Oncology, Devita et al., Eds., 4 th Edition, Vol. 1, pp. 248-275 (1993).

[0330] Also included are EDG binders and ribonucleotide reductase inhibitors. As used herein, the term "EDG binder" refers to a class of immunosuppressants that modulate lymphocyte recirculation, such as FTY720. The term "ribonucleotide reductase inhibitor" refers to a pyrimidine or purine nucleoside analog, including, but not limited to, fludarabine and / or cytosine arabinoside (ara-C), 6-thioguanine, 5-fluorouracil, cladribine, 6-mercaptopurine (especially in combination with ara-C for ALL), and / or pentostatin. Ribonucleotide reductase inhibitors are, in particular, hydroxyurea or 2-hydroxy-1H-isoindole-1,3-dione derivatives.

[0331] Also included are, inter alia, those compounds, proteins, or monoclonal antibodies of VEGF, such as 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine or a pharmaceutically acceptable salt thereof, 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine succinate; Angiostatin™; Endostatin™; anthranilic acid amide; ZD4190; ZD6474; SU5416; SU6668; bevacizumab; or anti-VEGF antibodies or anti-VEGF receptor antibodies, such as rhuMAb and RHUFab, VEGF aptamers, such as Macugon; FLT-4 inhibitors, FLT-3 inhibitors, VEGFR-2 IgGI antibodies, Angiozyme (RPI 4610), and bevacizumab (Avastin™).

[0332] As used herein, photodynamic therapy refers to therapy that uses certain chemicals known as photosensitizing compounds to treat or prevent cancer. Examples of photodynamic therapy include treatment with compounds such as Visudyne™ and porfimer sodium.

[0333] As used herein, angiogenic antisteroids refer to compounds that block or inhibit angiogenesis, such as anecortave, triamcinolone, hydrocortisone, 11-α-epihydrocortisol, cortexolone, 17α-hydroxyprogesterone, corticosterone, desoxycorticosterone, testosterone, estrone, and dexamethasone.

[0334] Corticosteroid-containing implants refer to compounds such as fluocinolone and dexamethasone.

[0335] Other chemotherapeutic compounds include, but are not limited to, plant alkaloids, hormonal compounds and antagonists; biological response modifiers, preferably lymphokines or interferons; antisense oligonucleotides or oligonucleotide derivatives; shRNA or siRNA; or miscellaneous compounds or compounds with other or unknown mechanisms of action.

[0336] The compounds of the present invention are also useful as co-therapeutic compounds for use in combination with other drug substances, such as anti-inflammatory, bronchodilatory, or antihistamine drug substances, particularly in the treatment of obstructive or inflammatory airway diseases such as those described above, for example, as enhancers of the therapeutic activity of such drugs or as a means of reducing the required dosing or potential side effects of such drugs. The compounds of the present invention can be mixed with the other drug substances in a fixed pharmaceutical composition, or they can be administered separately from, before, simultaneously with, or after the other drug substances. Thus, the present invention includes combinations of the compounds of the present invention described above with anti-inflammatory, bronchodilatory, antihistamine, or antitussive drug substances, wherein the compounds of the present invention and the drug substances are in the same or different pharmaceutical compositions.

[0337] Suitable anti-inflammatory drugs include steroids, in particular glucocorticosteroids such as budesonide, beclamethasone dipropionate, fluticasone propionate, ciclesonide, or mometasone furoate; nonsteroidal glucocorticoid receptor agonists; LTB4 antagonists such as LY293111, CGS025019C, CP-195543, SC-53228, BIIL 284, ONO 4057, SB 209247; LTD4 antagonists such as montelukast and zafirlukast; PDE4 inhibitors such as cilomilast (Ariflo® GlaxoSmithKline), roflumilast (Byk Gulden), V-11294A (Napp), BAY19-8004 (Bayer), SCH-351591 (Schering-Plough), Allofilin (Almirall Prodesfarma), PD189659 / PD168787 (Parke-Davis), AWD-12-281 (Asta Medica), CDC-801 (Celgene), SeICID™, CC-10004 (Celgene), VM554 / UM565 (Vernalis), T-440 (Tanabe), KW-4490 (Kyowa Hakko Kogyo); A2a agonists; A2b antagonists; and beta-2 ardenoceptor agonists such as albuterol (salbutamol), metaproterenol, terbutaline, salmeterol, fenoterol, procaterol, and, in particular, formoterol and its pharmaceutically acceptable salts. Suitable bronchodilators include anticholinergic or antimuscarinic compounds, in particular ipratropium bromide, oxitropium bromide, tiotropium salts, and CHF 4226 (Chiesi), and glycopyrrolate.

[0338] Suitable antihistamine drug substances include cetirizine hydrochloride, acetaminophen, clemastine fumarate, promethazine, loratidine, desloratidine, diphenhydramine and fexofenadine hydrochloride, activastine, astemizole, azelastine, ebastine, epinastine, mizolastine, and tefenadine.

[0339] Another useful combination of compounds of the invention with anti-inflammatory agents is with antagonists of chemokine receptors, e.g., CCR-1, CCR-2, CCR-3, CCR-4, CCR-5, CCR-6, CCR-7, CCR-8, CCR-9, and CCR10, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, particularly CCR-5 antagonists such as the Schering-Plough antagonists SC-351125, SCH-55700, and SCH-D, and Takeda antagonists such as N-[[4-[[[6,7-dihydro-2-(4-methylphenyl)-5H-benzo-cyclohepten-8-yl]carbonyl]amino]phenyl]-methyl]tetrahydro-N,N-dimethyl-2H-pyran-4-aminium chloride (TAK-770).

[0340] The structures of active compounds identified by code number, generic name or trade name can be obtained from the actual editions or databases of the standard compendium "The Merck Index", for example from international patents (e.g. IMS World Publications).

[0341] Exemplary Immuno-Oncology Agents In some embodiments, the one or more other therapeutic agents are immuno-oncology agents. As used herein, the term "immuno-oncology agent" refers to an agent that is effective in enhancing, stimulating, and / or upregulating the immune response in a subject. In some embodiments, the administration of an immuno-oncology agent with a compound of the present invention has a synergistic effect in the treatment of cancer.

[0342] The immuno-oncology agent can be, for example, a small molecule drug, an antibody, or a biological molecule or small molecule. Examples of biological immuno-oncology agents include, but are not limited to, cancer vaccines, antibodies, and cytokines. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the monoclonal antibody is humanized or human.

[0343] In some embodiments, the immuno-oncology agent is either (i) an agonist of a stimulatory (including costimulatory) receptor or (ii) an antagonist of an inhibitory (including costimulatory) signal on T cells, either of which results in amplification of antigen-specific T cell responses.

[0344] Certain stimulatory and inhibitory molecules are members of the immunoglobulin superfamily (IgSF). One important family of membrane-bound ligands that bind to costimulatory or co-inhibitory receptors is the B7 family, which includes B7-1, B7-2, B7-H1 (PD-L1), B7-DC (PD-L2), B7-H2 (ICOS-L), B7-H3, B7-H4, B7-H5 (VISTA), and B7-H6. Another family of membrane-bound ligands that bind to costimulatory or co-inhibitory receptors is the TNF family of molecules that bind to cognate TNF receptor family members, including CD40, and CD40L, OX-40, OX-40L, CD70, CD27L, CD30, CD30L, 4-1BBL, CD137 (4-1BB), TRAIL / Apo2-L, TRAILR1 / DR4, TRAILR2 / DR5, TRAILR3, TRAILR4, OPG, and RA. NK, RANKL, TWEAKR / Fn14, TWEAK, BAFFR, EDAR, XEDAR, TACI, APRIL, BCMA, LTβR, LIGHT, DcR3, HVEM, VEGI / TL1A, TRAMP / DR3 , EDAR, EDA1, XEDAR, EDA2, TNFR1, lymphotoxin α / TNFβ, TNFR2, TNFα, LTβR, lymphotoxin α1β2, FAS, FASL, RELT, DR6, TROY, NGFR.

[0345] In some embodiments, the immuno-oncology agent is a cytokine that inhibits T cell activation (e.g., IL-6, IL-10, TGF-β, VEGF, and other immunosuppressive cytokines) or a cytokine that stimulates T cell activation to stimulate an immune response.

[0346] In some embodiments, the combination of a compound of the present invention with an immuno-oncology agent can stimulate a T cell response. In some embodiments, the immuno-oncology agent is (i) an antagonist of a protein that inhibits T cell activation (e.g., an immune checkpoint inhibitor), such as CTLA-4, PD-1, PD-L1, PD-L2, LAG-3, TIM-3, Galectin-9, CEACAM-1, BTLA, CD69, Galectin-1, TIGIT, CD113, GPR56, VISTA, 2B4, CD48, GARP, PD1H, LAIR1, TIM-1, and TIM-4, or (ii) an agonist of a protein that stimulates T cell activation, such as B7-1, B7-2, CD28, 4-1BB (CD137), 4-1BBL, ICOS, ICOS-L, OX40, OX40L, GITR, GITRL, CD70, CD27, CD40, DR3, and CD28H.

[0347] In some embodiments, the immuno-oncology agent is an antagonist of an inhibitory receptor on NK cells or an agonist of an activating receptor on NK cells, hi some embodiments, the immuno-oncology agent is an antagonist of a KIR, such as lirilumab.

[0348] In some embodiments, the immuno-oncology agent is an agent that inhibits or depletes macrophages or monocytes, including, but not limited to, CSF-1R antagonists such as RG7155 (WO11 / 70024, WO11 / 107553, WO11 / 131407, WO13 / 87699, WO13 / 119716, WO13 / 132044), or CSF-1R antagonist antibodies, including FPA-008 (WO11 / 140249, WO13169264, WO14 / 036357).

[0349] In some embodiments, the immuno-oncology agent is selected from agonist agents that ligate positive costimulatory receptors, blocking agents that attenuate signaling through inhibitory receptors, antagonists, and one or more agents that systemically increase the frequency of anti-tumor T cells, agents that overcome different immunosuppressive pathways within the tumor microenvironment (e.g., blocking inhibitory receptor engagement (e.g., PD-L1 / PD-1 interaction), depleting or inhibiting Tregs (e.g., using anti-CD25 monoclonal antibodies (e.g., daclizumab) or by ex vivo anti-CD25 bead depletion), inhibiting metabolic enzymes such as IDO, or reversing / preventing T cell energy or exhaustion), and agents that induce innate immune activation and / or inflammation at the tumor site.

[0350] In some embodiments, the immuno-oncology agent is a CTLA-4 antagonist. In some embodiments, the CTLA-4 antagonist is an antagonistic CTLA-4 antibody. In some embodiments, the antagonistic CTLA-4 antibody is YERVOY (ipilimumab) or tremelimumab.

[0351] In some embodiments, the immuno-oncology agent is a PD-1 antagonist. In some embodiments, the PD-1 antagonist is administered by infusion. In some embodiments, the immuno-oncology agent is an antibody or antigen-binding portion thereof that specifically binds to the programmed death-1 (PD-1) receptor and inhibits PD-1 activity. In some embodiments, the PD-1 antagonist is an antagonistic PD-1 antibody. In some embodiments, the antagonistic PD-1 antibody is OPDIVO (nivolumab), KEYTRUDA (pembrolizumab), or MEDI-0680 (AMP-514; WO 2012 / 145493). In some embodiments, the immuno-oncology agent can be pidilizumab (CT-011). In some embodiments, the immuno-oncology agent is a recombinant protein called AMP-224, which is composed of the extracellular domain of PD-L2 (B7-DC) fused to the Fc portion of IgG1.

[0352] In some embodiments, the immuno-oncology agent is a PD-L1 antagonist. In some embodiments, the PD-L1 antagonist is an antagonistic PD-L1 antibody. In some embodiments, the PD-L1 antibody is MPDL3280A (RG7446; WO2010 / 077634), durvalumab (MEDI4736), BMS-936559 (WO2007 / 005874), and MSB0010718C (WO2013 / 79174).

[0353] In some embodiments, the immuno-oncology agent is a LAG-3 antagonist. In some embodiments, the LAG-3 antagonist is an antagonistic LAG-3 antibody. In some embodiments, the LAG-3 antibody is BMS-986016 (WO10 / 19570, WO14 / 08218), or IMP-731 or IMP-321 (WO08 / 132601, WO009 / 44273).

[0354] In some embodiments, the immuno-oncology agent is a CD137 (4-1BB) agonist. In some embodiments, the CD137 (4-1BB) agonist is an agonistic CD137 antibody. In some embodiments, the CD137 antibody is urelumab or PF-05082566 (WO12 / 32433).

[0355] In some embodiments, the immuno-oncology agent is a GITR agonist. In some embodiments, the GITR agonist is an agonistic GITR antibody. In some embodiments, the GITR antibody is BMS-986153, BMS-986156, TRX-518 (WO006 / 105021, WO009 / 009116), or MK-4166 (WO11 / 028683).

[0356] In some embodiments, the immuno-oncology agent is an indoleamine (2,3)-dioxygenase (IDO) antagonist. In some embodiments, the IDO antagonist is selected from epacadostat (INCB024360, Incyte), indoximod (NLG-8189, NewLink Genetics Corporation), capmanitib (INC280, Novartis), GDC-0919 (Genentech / Roche), PF-06840003 (Pfizer), BMS:F001287 (Bristol-Myers Squibb), Phy906 / KD108 (Phytoceutica), an enzyme that breaks down kynurenine (Kynase, Ikena Oncology, formerly known as Kyn therapy), and NLG-919 (WO09 / 73620, WO009 / 1156652, WO11 / 56652, WO12 / 142237).

[0357] In some embodiments, the immuno-oncology agent is an OX40 agonist. In some embodiments, the OX40 agonist is an agonistic OX40 antibody. In some embodiments, the OX40 antibody is MEDI-6383 or MEDI-6469.

[0358] In some embodiments, the immuno-oncology agent is an OX40L antagonist. In some embodiments, the OX40L antagonist is an antagonistic OX40 antibody. In some embodiments, the OX40L antagonist is RG-7888 (WO06 / 029879).

[0359] In some embodiments, the immuno-oncology agent is a CD40 agonist. In some embodiments, the CD40 agonist is an agonistic CD40 antibody. In some embodiments, the immuno-oncology agent is a CD40 antagonist. In some embodiments, the CD40 antagonist is an antagonistic CD40 antibody. In some embodiments, the CD40 antibody is lucatumumab or dacetuzumab.

[0360] In some embodiments, the immuno-oncology agent is a CD27 agonist. In some embodiments, the CD27 agonist is an agonist CD27 antibody. In some embodiments, the CD27 antibody is varlilumab.

[0361] In some embodiments, the immuno-oncology agent is MGA271 (directed against B7H3) (WO11 / 109400).

[0362] In some embodiments, the immuno-oncology agent is abagovomab, adecatumumab, afutuzumab, alemtuzumab, anatumomab mafenatox, apolizumab, atezolimab, avelumab, blinatumomab, BMS-936559, catumaxomab, durvalumab, epacadostat, epratuzumab, indoximod, inotuzumab ozogamicin, intelumab, ipilimumab, isaximab, lambrolizumab, MED14736, MPDL3280A, nivolumab, obinutuzumab, ocaratuzumab, ofatumumab, olattumumab, pembrolizumab, pidilizumab, rituximab, ticilimumab, samalizumab, or tremelimumab.

[0363] In some embodiments, the immuno-oncology agent is an immunostimulatory agent. For example, antibodies that block the PD-1 and PD-L1 inhibitory axis can release activated tumor-reactive T cells and have been shown in clinical trials to induce durable anti-tumor responses in an increasing number of tumor tissue types, including some tumor types not previously considered sensitive to immunotherapy. See, e.g., Okazaki, T. et al. (2013) Nat. Immunol. 14, 1212-1218; Zou et al. (2016) Sci. Transl. Med. 8. The anti-PD-1 antibody nivolumab (Opdivo®, Bristol-Myers Squibb, also known as ONO-4538, MDX1106, and BMS-936558) has shown the potential to improve overall survival in RCC patients who have experienced disease progression during or after previous antiangiogenic therapy.

[0364] In some embodiments, the immunomodulatory therapeutic agent specifically induces apoptosis of tumor cells. Approved immunomodulatory therapeutic agents that may be used in the present invention include pomalidomide (Pomalyst®, Celgene), lenalidomide (Revlimid®, Celgene), and ingenol mebutate (Picato®, LEO Pharma).

[0365] In some embodiments, the immuno-oncology agent is a cancer vaccine. In some embodiments, the cancer vaccine is selected from sipuleucel-T (Provenge®, Dendreon / Valeant Pharmaceuticals), which is approved for the treatment of asymptomatic or minimally symptomatic metastatic castration-resistant (hormone-refractory) prostate cancer, and talimogene laherparepvec (Imlygic®, BioVex / Amgen, formerly known as T-VEC), a recombinant oncolytic virotherapy approved for the treatment of unresectable cutaneous, subcutaneous, and nodular melanoma. In some embodiments, the immuno-oncology agent is an oncolytic virotherapy, e.g., pexastimogene devacirepvec (PexaVec / JX-594, SillaJen / formerly Jennerex). Biotherapeutics, a thymidine kinase (TK)-deficient vaccinia virus engineered to express GM-CSF for hepatocellular carcinoma (NCT02562755) and melanoma (NCT00429312); and Reolysin® (Oncolytics), a thymidine kinase (TK)-deficient vaccinia virus engineered to express GM-CSF, which does not replicate in cells without RAS activation in many cancers, including colorectal cancer (NCT01622543), prostate cancer (NCT01619813), head and neck squamous cell carcinoma (NCT01166542), pancreatic adenocarcinoma (NCT00998322), and non-small cell lung cancer (NSCLC) (NCT00861627).Biotech), a variant of reovirus, enadenotucirev (NG-348, PsiOxus, formerly known as ColoAd1), an adenovirus engineered to express full-length CD80 and T-cell receptor CD3 proteins and specific antibody fragments in metastatic or advanced epithelial tumors such as ovarian cancer (NCT02028117), colorectal cancer, bladder cancer, head and neck squamous cell carcinoma, salivary gland cancer (NCT02636036), melanoma (NCT03003676), and peritoneal disease, colorectal cancer, or ONCOS-102 (Targovax / formerly Oncos), an adenovirus engineered to express GM-CSF in ovarian cancer (NCT02963831); GL-ONC1 (GLV-1h68 / GLV-1h153, Genelux GmbH) or CG0070 (Cold Genesys), vaccinia viruses engineered to express beta-galactosidase (beta-gal) / beta-glucoronidase or beta-gal / human sodium iodide symporter (hNIS), respectively, which have been studied in peritoneal carcinomatosis (NCT01443260), fallopian tube cancer, and ovarian cancer (NCT02759588); and an adenovirus engineered to express GM-CSF in bladder cancer (NCT02365818).

[0366] In some embodiments, the immuno-oncology agent is JX-929 (SillaJen / formerly Jennerex Biotherapeutics), a TK and vaccinia growth factor-deficient vaccinia virus engineered to express cytosine deaminase capable of converting the prodrug 5-fluorocytosine to the cytotoxic drug 5-fluorouracil; TG01 and TG02 (Targovax / formerly Oncos), peptide-based immunotherapeutics targeting hard-to-treat RAS mutations; TILT-123 (TILT Biotherapeutics), an engineered adenovirus called Ad5 / 3-E2F-delta24-hTNFα-IRES-hIL20; and TILT-123 (TILT Biotherapeutics), an engineered adenovirus engineered to express the glycoprotein (GP) of lymphocytic choriomeningitis virus (LCMV) and targeting antigen-specific CD8+ Selected from VSV-GP (ViraTherapeutics), a vesicular stomatitis virus (VSV) that can be further engineered to express antigens designed to elicit a T cell response.

[0367] In some embodiments, the immuno-oncology agent is a T cell engineered to express a chimeric antigen receptor, or CAR. Such T cells engineered to express a chimeric antigen receptor are called CAR-T cells.

[0368] CARs have been constructed that consist of a binding domain derived from a natural ligand, a single-chain variable fragment (scFv), derived from a monoclonal antibody specific for a cell surface antigen fused to an endodomain, the functional end of a T cell receptor (TCR), such as the CD3-zeta signaling domain from the TCR, which can generate an activation signal in T lymphocytes. Upon antigen binding, such CARs couple to endogenous signaling pathways in effector cells and generate activation signals similar to those initiated by the TCR complex.

[0369] For example, in some embodiments, the CAR-T cells are one of those described in U.S. Pat. No. 8,906,682 (June et al.; incorporated herein by reference in its entirety), which discloses CAR-T cells engineered to include an extracellular domain bearing an antigen-binding domain (e.g., a domain that binds to CD19) fused to the intracellular signaling domain of the T-cell antigen receptor complex zeta chain (e.g., CD3 zeta). When expressed in T cells, the CAR can redirect antigen recognition based on antigen-binding specificity. In the case of CD19, the antigen is expressed on malignant B cells. Currently, there are over 200 clinical trials underway employing CAR-T for a wide range of indications. [https: / / clinicaltrials.gov / ct2 / results?term=chimeric+antigen+receptors&pg=1]

[0370] In some embodiments, the immunostimulatory agent is an activator of retinoic acid receptor-related orphan receptor gamma (RORγt). RORγt is a transcription factor that plays a key role in the differentiation and maintenance of type 17 effector subsets of CD4+ (Th17) and CD8+ (Tc17) T cells, as well as in the differentiation of IL-17 expressing innate immune cell subpopulations such as NK cells. In some embodiments, the activator of RORγt is LYC-55716 (Lycera), which is currently being evaluated in clinical trials for the treatment of solid tumors (NCT02929862).

[0371] In some embodiments, the immunostimulatory agent is a toll-like receptor (TLR) agonist or activator. Suitable TLR activators include TLR9 agonists or activators, such as SD-101 (Dynavax). SD-101 is an immunostimulatory CpG that is being studied in B-cell, follicular lymphoma, and other lymphomas (NCT02254772). TLR8 agonists or activators that can be used in the present invention include motolimod (VTX-2337, VentiRx Pharmaceuticals), which is being studied in squamous cell carcinoma of the head and neck (NCT02124850) and ovarian cancer (NCT02431559).

[0372] Other immuno-oncology agents that can be used in the present invention include urelumab (BMS-663513, Bristol-Myers Squibb), an anti-CD137 monoclonal antibody, valilumab (CDX-1127, Celldex Therapeutics), an anti-CD27 monoclonal antibody, BMS-986178 (Bristol-Myers Squibb), an anti-OX40 monoclonal antibody, lirilumab (IPH2102 / BMS-986015, Innate Pharma, Bristol-Myers Squibb), an anti-KIR monoclonal antibody, monalizumab (IPH2201, Innate Pharma, AstraZeneca), an anti-NKG2A monoclonal antibody, andecaliximab (GS-5745, Gilead Sciences), an anti-MMP9 antibody, MK-4166 (Merck & Co.), and an anti-GITR monoclonal antibody.

[0373] In some embodiments, the immunostimulatory agent is selected from elotuzumab, mifamurtide, agonists or activators of toll-like receptors, and activators of RORγt.

[0374] In some embodiments, the immune stimulatory therapy is recombinant human interleukin-15 (rhIL-15). rhIL-15 is being clinically tested as a therapy for melanoma and renal cell carcinoma (NCT01021059 and NCT01369888) and leukemia (NCT02689453). In some embodiments, the immune stimulatory agent is recombinant human interleukin-12 (rhIL-12). In some embodiments, the IL-15-based immunotherapeutic is heterodimeric IL-15 (hetIL-15, Novartis / Admune), a fusion complex composed of a synthetic form of endogenous IL-15 complexed with the soluble IL-15 binding protein IL-15 receptor alpha chain (IL15:sIL-15RA), which is being tested in Phase 1 clinical trials in melanoma, renal cell carcinoma, non-small cell lung cancer, and head and neck squamous cell carcinoma (NCT02452268). In some embodiments, the recombinant human interleukin-12 (rhIL-12) is NM-IL-12 (Neumedicines, Inc.), NCT02544724, or NCT02542124.

[0375] In some embodiments, the immuno-oncology agent is selected from those described in Jerry L. Adams et al., "Big opportunities for small molecules in immuno-oncology," Cancer Therapy 2015, Vol. 14, pp. 603-622, the entire contents of which are incorporated herein by reference. In some embodiments, the immuno-oncology agent is selected from the examples described in Table 1 of Jerry L. Adams et al. In some embodiments, the immuno-oncology agent is a small molecule that targets an immune-oncology target selected from those listed in Table 2 of L. Adams et al. In some embodiments, the immuno-oncology agent is a small molecule agent selected from those listed in Table 2 of Jerry L. Adams et al.

[0376] In some embodiments, the immuno-oncology agent is selected from the small molecule immuno-oncology agents described in Peter L. Toogood, "Small molecule immuno-oncology therapeutic agents," Bioorganic & Medicinal Chemistry Letters 2018, Vol. 28, pp. 319-329, the entire contents of which are incorporated herein by reference. In some embodiments, the immuno-oncology agent is an agent that targets a pathway described in Peter L. Toogood.

[0377] In some embodiments, the immuno-oncology agent is selected from those described in Sandra L. Ross et al., "Bispecific T cell engager (BiTE®) antibody constructs can mediate bystander tumor cell killing," PLoS ONE 12(8):e0183390, the contents of which are incorporated herein by reference in their entirety. In some embodiments, the immuno-oncology agent is a bispecific T cell engager (BiTE®) antibody construct. In some embodiments, the bispecific T cell engager (BiTE®) antibody construct is a CD19 / CD3 bispecific antibody construct. In some embodiments, the bispecific T cell engager (BiTE®) antibody construct is an EGFR / CD3 bispecific antibody construct. In some embodiments, the bispecific T cell engager (BiTE®) antibody construct activates T cells. In some embodiments, the bispecific T cell engager (BiTE®) antibody construct activates T cells and releases cytokines that induce upregulation of intercellular adhesion molecule 1 (ICAM-1) and FAS on bystander cells. In some embodiments, the bispecific T cell engager (BiTE®) antibody construct activates T cells, resulting in the induction of bystander cell lysis. In some embodiments, the bystander cells are within a solid tumor. In some embodiments, the bystander cells to be lysed are in proximity to the BiTE®-activated T cells. In some embodiments, the bystander cells comprise tumor-associated antigen (TAA)-negative cancer cells. In some embodiments, the bystander cells comprise EGFR-negative cancer cells. In some embodiments, the immuno-oncology agent is an antibody that blocks the PD-L1 / PD1 axis and / or CTLA4. In some embodiments, the immuno-oncology agent is ex vivo expanded tumor-infiltrating T cells. In some embodiments, the immuno-oncology agent is a bispecific antibody construct or a chimeric antigen receptor (CAR) that directly links T cells to tumor-associated surface antigens (TAA).

[0378] Exemplary Immune Checkpoint Inhibitors In some embodiments, the immuno-oncology agent is an immune checkpoint inhibitor described herein.

[0379] As used herein, the term "checkpoint inhibitor" refers to a drug that is useful for preventing cancer cells from evading the patient's immune system.One of the main mechanisms of anti-tumor immune destruction is known as "T cell exhaustion", which is caused by chronic exposure to antigens, which leads to the upregulation of inhibitory receptors.These inhibitory receptors function as immune checkpoints to prevent uncontrolled immune responses.

[0380] PD-1 and co-inhibitory receptors, such as cytotoxic T lymphocyte antigen 4 (CTLA-4), B- and T-lymphocyte attenuator (BTLA; CD272), T-cell immunoglobulin and mucin domain-3 (Tim-3), lymphocyte activation gene-3 (Lag-3; CD223), and others, are often referred to as checkpoint regulators. They act as molecular "gatekeepers" that allow extracellular signals to determine whether cell cycle progression and other intracellular signaling processes should proceed.

[0381] In some embodiments, the immune checkpoint inhibitor is an antibody against PD-1, which binds to the programmed cell death 1 receptor (PD-1) and prevents the receptor from binding to the inhibitory ligand PDL-1, thus abolishing the tumor's ability to suppress the host anti-tumor immune response.

[0382] In some embodiments, the checkpoint inhibitor is a biotherapeutic or a small molecule. In some embodiments, the checkpoint inhibitor is a monoclonal antibody, a humanized antibody, a fully human antibody, a fusion protein, or a combination thereof. In some embodiments, the checkpoint inhibitor inhibits a checkpoint protein selected from CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, B-7 family ligand, or a combination thereof. In some embodiments, the checkpoint inhibitor interacts with a ligand of a checkpoint protein selected from CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, a B-7 family ligand, or a combination thereof. In some embodiments, the checkpoint inhibitor is an immunostimulant, a T cell growth factor, an interleukin, an antibody, a vaccine, or a combination thereof. In some embodiments, the interleukin is IL-7 or IL-15. In some embodiments, the interleukin is glycosylated IL-7. In an additional aspect, the vaccine is a dendritic cell (DC) vaccine.

[0383] Checkpoint inhibitors include any agent that blocks or inhibits an inhibitory pathway of the immune system in a statistically significant manner. Such inhibitors may include small molecule inhibitors, or may include antibodies or antigen-binding fragments thereof that bind to and block or inhibit immune checkpoint receptors, or antibodies that bind to and block or inhibit immune checkpoint receptor ligands. Exemplary checkpoint molecules that can be targeted for blockade or inhibition include CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, GAL9, LAG3, TIM3, VISTA, KIR, 2B4 (which belong to the CD2 family of molecules and are involved in NK, gamma delta, and memory CD8+ (expressed on all αβ T cells), CD160 (also known as BY5), CGEN-15049, CHK1 and CHK2 kinases, A2aR, and various B-7 family ligands. B7 family ligands include, but are not limited to, B7-1, B7-2, B7-DC, B7-H1, B7-H2, B7-H3, B7-H4, B7-H5, B7-H6, and B7-H7. Checkpoint inhibitors include antibodies, or antigen-binding fragments thereof, other binding proteins, biotherapeutics, or small molecules that bind to and block or inhibit the activity of one or more of CTLA-4, PDL1, PDL2, PD1, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, and CGEN-15049. Exemplary immune checkpoint inhibitors include, but are not limited to, tremelimumab (a CTLA-4 blocking antibody), anti-OX40, PD-L1 monoclonal antibody (anti-B7-H1; MEDI4736), MK-3475 (a PD-1 blocker), nivolumab (an anti-PD1 antibody), CT-011 (an anti-PD1 antibody), BY55 monoclonal antibody, AMP224 (an anti-PDL1 antibody), BMS-936559 (an anti-PDL1 antibody), MPLDL3280A (an anti-PDL1 antibody), MSB0010718C (an anti-PDL1 antibody), and ipilimumab (an anti-CTLA-4 checkpoint inhibitor). Checkpoint protein ligands include, but are not limited to, PD-L1, PD-L2, B7-H3, B7-H4, CD28, CD86, and TIM-3.

[0384] In certain embodiments, the immune checkpoint inhibitor is selected from a PD-1 antagonist, a PD-L1 antagonist, and a CTLA-4 antagonist. In some embodiments, the checkpoint inhibitor is selected from the group consisting of nivolumab (Opdivo®), ipilimumab (Yervoy®), and pembrolizumab (Keytruda®). In some embodiments, the checkpoint inhibitor is selected from nivolumab (anti-PD-1 antibody, Opdivo®, Bristol-Myers Squibb), pembrolizumab (anti-PD-1 antibody, Keytruda®, Merck), ipilimumab (anti-CTLA-4 antibody, Yervoy®, Bristol-Myers Squibb), durvalumab (anti-PD-L1 antibody, Imfinzi®, AstraZeneca), and atezolizumab (anti-PD-L1 antibody, Tecentriq®, Genentech).

[0385] In some embodiments, the checkpoint inhibitor is selected from the group consisting of lambrolizumab (MK-3475), nivolumab (BMS-936558), pidilizumab (CT-011), AMP-224, MDX-1105, MEDI4736, MPDL3280A, BMS-936559, ipilimumab, rilumab, IPH2101, pembrolizumab (Keytruda®), and tremelimumab.

[0386] In some embodiments, the immune checkpoint inhibitor is REGN2810 (Regeneron), an anti-PD-1 antibody studied in patients with basal cell carcinoma (NCT03132636), NSCLC (NCT03088540), cutaneous squamous cell carcinoma (NCT02760498), lymphoma (NCT02651662), and melanoma (NCT03002376); pidilizumab (CureTech), also known as CT-011, an antibody that binds PD-1 in clinical trials for diffuse large B-cell lymphoma and multiple myeloma; and avelumab (Bavencio®, Pfizer / Merck), also known as MSB0010718C, a fully human IgG1 anti-PD-L1 antibody in clinical trials for non-small cell lung cancer, Merkel cell carcinoma, mesothelioma, solid tumors, kidney cancer, ovarian cancer, bladder cancer, head and neck cancer, and gastric cancer. KGaA), or PDR001 (Novartis), a blocking antibody that binds to PD-1 in clinical trials for non-small cell lung cancer, melanoma, triple-negative breast cancer, and advanced or metastatic solid tumors. Tremelimumab (CP-675,206; Astrazeneca) is a fully human monoclonal antibody against CTLA-4 that is being studied in clinical trials for many indications, including mesothelioma, colorectal cancer, renal cancer, breast cancer, lung and non-small cell lung cancer, pancreatic ductal adenocarcinoma, pancreatic cancer, germ cell cancer, squamous cell carcinoma of the head and neck, hepatocellular carcinoma, prostate cancer, endometrial cancer, metastatic cancer of the liver, liver cancer, large B-cell lymphoma, ovarian cancer, cervical cancer, metastatic anaplastic thyroid cancer, urothelial carcinoma, fallopian tube cancer, multiple myeloma, bladder cancer, soft tissue sarcoma, and melanoma. AGEN-1884 (Agenus) is an anti-CTLA4 antibody being studied in a Phase 1 clinical trial in advanced solid tumors (NCT02694822).

[0387] In some embodiments, the checkpoint inhibitor is an inhibitor of T-cell immunoglobulin mucin-containing protein 3 (TIM-3). TIM-3 inhibitors that can be used in the present invention include TSR-022, LY3321367, and MBG453. TSR-022 (Tesaro) is an anti-TIM-3 antibody being studied in solid tumors (NCT02817633). LY3321367 (Eli Lilly) is an anti-TIM-3 antibody being studied in solid tumors (NCT03099109). MBG453 (Novartis) is an anti-TIM-3 antibody being studied in advanced malignancies (NCT02608268).

[0388] In some embodiments, the checkpoint inhibitor is an inhibitor of TIGIT, a T cell immunoreceptor with Ig and ITIM domains or an immunoreceptor on certain T cells and NK cells. TIGIT inhibitors that can be used in the present invention include BMS-986207 (Bristol-Myers Squibb), anti-TIGIT monoclonal antibody (NCT02913313), OMP-313M32 (Oncomed), and anti-TIGIT monoclonal antibody (NCT03119428).

[0389] In some embodiments, the checkpoint inhibitor is an inhibitor of lymphocyte activation gene 3 (LAG-3). LAG-3 inhibitors that can be used in the present invention include BMS-986016, REGN3767, and IMP321. BMS-986016 (Bristol-Myers Squibb), an anti-LAG-3 antibody, is being studied in glioblastoma and gliosarcoma (NCT02658981). REGN3767 (Regeneron) is also an anti-LAG-3 antibody and is being studied in malignant tumors (NCT03005782). IMP321 (Immutep SA), a LAG-3-Ig fusion protein, is being studied in melanoma (NCT02676869), adenocarcinoma (NCT02614833), and metastatic breast cancer (NCT00349934).

[0390] Checkpoint inhibitors that can be used in the present invention include OX40 agonists. OX40 agonists being studied in clinical trials include PF-04518600 / PF-8600 (Pfizer), an agonist anti-OX40 antibody in metastatic renal cancer (NCT03092856) and advanced cancers and neoplasms (NCT02554812; NCT05082566), GSK3174998 (Merck), an agonist anti-OX40 antibody in a Phase 1 cancer trial (NCT02528357), and MEDI, an agonist anti-OX40 antibody in advanced solid tumors (NCT02318394 and NCT02705482). 0562 (Medimmune / AstraZeneca), MEDI6469, an agonist anti-OX40 antibody (Medimmune / AstraZeneca) in patients with colorectal cancer (NCT02559024), breast cancer (NCT01862900), head and neck cancer (NCT02274155), and metastatic prostate cancer (NCT01303705), and BMS-986178 (Bristol-Myers Squibb), an agonist anti-OX40 antibody in advanced cancers (NCT02737475).

[0391] Checkpoint inhibitors that may be used in the present invention include CD137 (also known as 4-1BB) agonists. CD137 agonists being studied in clinical trials include utomilumab (PF-05082566, Pfizer), an agonist anti-CD137 antibody in diffuse large B-cell lymphoma (NCT02951156) and advanced cancers and neoplasms (NCT02554812 and NCT05082566); urelumab (BMS-663513, Bristol-Myers Squibb), an agonist anti-CD137 antibody in melanoma and skin cancer (NCT02652455) and glioblastoma and gliosarcoma (NCT02658981); and CTX-471 (Compass Therapeutics), an agonist anti-CD137 antibody in metastatic or locally advanced malignancies (NCT03881488).

[0392] Checkpoint inhibitors that can be used in the present invention include CD27 agonists. CD27 agonists being studied in clinical trials include varlilumab (CDX-1127, Celldex Therapeutics), an agonistic anti-CD27 antibody in squamous cell head and neck cancer, ovarian cancer, colorectal cancer, renal cell carcinoma, and glioblastoma (NCT02335918), lymphoma (NCT01460134), and glioma and astrocytoma (NCT02924038).

[0393] Checkpoint inhibitors that can be used in the present invention include glucocorticoid-induced tumor necrosis factor receptor (GITR) agonists. GITR agonists being studied in clinical trials include TRX518 (Leap Therapeutics), an agonist anti-GITR antibody in melanoma and other malignant solid tumors (NCT01239134 and NCT02628574), GWN323 (Novartis), an agonist anti-GITR antibody in solid tumors and lymphomas (NCT02740270), INCAGN01876 (Incyte / Agenus), an agonist anti-GITR antibody in advanced cancers (NCT02697591 and NCT03126110), MK-4166 (Merck), an agonist anti-GITR antibody in solid tumors (NCT02132754), and human IgG1 (Merck) in advanced solid tumors. and MEDI1873 (Medimmune / AstraZeneca), an agonistic hexameric GITR ligand molecule with an Fc domain (NCT02583165).

[0394] Checkpoint inhibitors that can be used in the present invention include inducible T cell costimulatory factor (ICOS, also known as CD278) agonists. ICOS agonists being studied in clinical trials include MEDI-570 (Medimmune), an agonist anti-ICOS antibody in lymphoma (NCT02520791), GSK3359609 (Merck), an agonist anti-ICOS antibody in phase 1 (NCT02723955), and JTX-2011 (Jounce Therapeutics), an agonist anti-ICOS antibody in phase 1 (NCT02904226).

[0395] Checkpoint inhibitors that may be used in the present invention include killer IgG-like receptor (KIR) inhibitors. KIR inhibitors being studied in clinical trials include lirilumab (IPH2102 / BMS-986015, Innate Pharma / Bristol-Myers Squibb), an anti-KIR antibody in leukemia (NCT01687387, NCT02399917, NCT02481297, NCT02599649), multiple myeloma (NCT02252263), and lymphoma (NCT01592370); IPH2101 (1-7F9, Innate Pharma) in myeloma (NCT01222286 and NCT01217203); and IPH4102 (Innate Pharma), an anti-KIR antibody that binds to the three domains of the long cytoplasmic tail (KIR3DL2) in lymphoma (NCT02593045).

[0396] Checkpoint inhibitors that can be used in the present invention include CD47 inhibitors of the interaction between CD47 and signal regulatory protein alpha (SIRPa). CD47 / SIRPa inhibitors being studied in clinical trials include ALX-148 (Alexo Therapeutics), an antagonistic variant of (SIRPa) that binds to CD47 and inhibits CD47 / SIRPa-mediated signaling in Phase 1 clinical trials (NCT03013218), and TTI-621 (SIRPa-Fc, Trillium), a soluble recombinant fusion protein created by linking the N-terminal CD47-binding domain of SIRPa with the Fc domain of human IgG1, which acts by binding to human CD47 and preventing it from sending a "no phagocytosis" signal to macrophages in Phase 1 clinical trials (NCT02890368 and NCT02663518). Therapeutics), CC-90002 (Celgene), an anti-CD47 antibody in leukemia (NCT02641002), and Hu5F9-G4 (Forty Seven, Inc.) in colorectal neoplasia and solid tumors (NCT02953782), acute myeloid leukemia (NCT02678338), and lymphoma (NCT02953509).

[0397] Checkpoint inhibitors that can be used in the present invention include CD73 inhibitors. CD73 inhibitors being studied in clinical trials include MEDI9447 (Medimmune), an anti-CD73 antibody in solid tumors (NCT02503774), and BMS-986179 (Bristol-Myers Squibb), an anti-CD73 antibody in solid tumors (NCT02754141).

[0398] Checkpoint inhibitors that can be used in the present invention include agonists of stimulator of interferon genes (STING, also known as transmembrane protein 173, or TMEM173). STING agonists being studied in clinical trials include MK-1454 (Merck), an agonist synthetic cyclic dinucleotide in lymphoma (NCT03010176), and ADU-S100 (MIW815, Aduro Biotech / Novartis), an agonist synthetic cyclic dinucleotide in Phase 1 trials (NCT02675439 and NCT03172936).

[0399] Checkpoint inhibitors that can be used in the present invention include CSF1R inhibitors. CSF1R inhibitors being studied in clinical trials include pexidartinib (PLX3397, Plexxikon), a CSF1R small molecule inhibitor in colorectal cancer, pancreatic cancer, metastatic and advanced cancers (NCT02777710), melanoma, non-small cell lung cancer, head and neck squamous cell carcinoma, gastrointestinal stromal tumor (GIST), and ovarian cancer (NCT02452424), as well as pancreatic cancer (NCT03153410), melanoma (NCT03101254). and IMC-CS4 (LY3022855, Lilly), an anti-CSF-1R antibody in solid tumors (NCT02718911), and BLZ945 (4-[2((1R,2R)-2-hydroxycyclohexylamino)-benzothiazol-6-yloxyl]-pyridine-2-carboxylic acid methylamide, Novartis), an orally available CSF1R inhibitor in advanced solid tumors (NCT02829723).

[0400] Checkpoint inhibitors that can be used in the present invention include NKG2A receptor inhibitors, which are being studied in clinical trials, including monalizumab (IPH2201, Innate Pharma), an anti-NKG2A antibody targeted at head and neck tumors (NCT02643550) and chronic lymphocytic leukemia (NCT02557516).

[0401] In some embodiments, the immune checkpoint inhibitor is selected from nivolumab, pembrolizumab, ipilimumab, avelumab, durvalumab, atezolizumab, or pidilizumab.

[0402] The compounds of the present invention may also be used in combination with known therapeutic processes, such as the administration of hormones or radiation. In certain embodiments, provided compounds are used as radiosensitizers, particularly for the treatment of tumors that exhibit poor sensitivity to radiation therapy.

[0403] The compounds of the present invention can be administered alone or in combination with one or more other therapeutic compounds, and possible combination therapy takes the form of a fixed combination, or the administration of the compounds of the present invention and one or more other therapeutic compounds that are given alternately or independently, or the combined administration of a fixed combination and one or more other therapeutic compounds.The compounds of the present invention can also be administered in combination with chemotherapy, radiotherapy, immunotherapy, phototherapy, surgical intervention, or a combination thereof, particularly for tumor therapy, or in addition.Long-term therapy is possible in the context of other treatment strategies, as described above, as well as adjuvant therapy.Other possible treatments are therapy to maintain the patient's condition after tumor regression, or further, for example, chemoprevention therapy in at-risk patients.

[0404] These additional agents can be administered separately from the composition containing the compound of the present invention as part of a multiple dose regimen. Alternatively, these agents can be part of a single dosage form, mixed together with the compound of the present invention in a single composition. When administered as part of a multiple dose regimen, the two active agents can be administered simultaneously, sequentially, or within a certain period of each other, usually within 5 hours of each other.

[0405] As used herein, the terms "combination," "in combination," and related terms refer to simultaneous or sequential administration of therapeutic agents according to the invention. For example, a compound of the invention may be administered with another therapeutic agent, either simultaneously or sequentially, in separate unit dosage forms or together in a single unit dosage form. Thus, the invention provides a single unit dosage form comprising a compound of the invention, an additional therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

[0406] The amounts of both the compounds of the present invention and additional therapeutic agents (in compositions containing additional therapeutic agents described above) that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. Preferably, the compositions of the present invention should be formulated so that a dosage of 0.01 to 100 mg / kg body weight / day of the compounds of the present invention can be administered.

[0407] In compositions containing an additional therapeutic agent, the additional therapeutic agent and the compound of the present invention may act synergistically. Thus, the amount of additional therapeutic agent in such compositions will be less than the amount required in a monotherapy utilizing only that therapeutic agent. In such compositions, a dosage of 0.01 to 1,000 μg / kg body weight / day of the additional therapeutic agent can be administered.

[0408] The amount of additional therapeutic agent present in the compositions of the invention is no more than the amount that would normally be administered in a composition comprising that therapeutic agent as the only active agent. Preferably, the amount of additional therapeutic agent in the compositions of the present disclosure ranges from about 50% to 100% of the amount that would normally be present in a composition comprising that agent as the only therapeutically active agent.

[0409] The compounds of the present invention, or pharmaceutical compositions thereof, may also be incorporated into compositions for coating implantable medical devices, such as prostheses, artificial valves, artificial blood vessels, stents, and catheters. For example, vascular stents have been used to overcome restenosis (re-narrowing of the blood vessel wall after injury). However, patients using stents or other implantable devices are at risk of thrombus formation or platelet activation. These undesirable effects can be prevented or mitigated by pre-coating the device with a pharmaceutically acceptable composition containing a kinase inhibitor. An implantable device coated with the compounds of the present invention is another embodiment of the present invention. [Example]

[0410] As shown in the examples below, in certain exemplary embodiments, compounds are prepared according to the following general procedures. While the general methods illustrate the synthesis of certain specific compounds of the invention, the following general methods, and other methods known to those of skill in the art, can be applied to all compounds as described herein and to each subclass and species of these compounds. Additional compounds of the invention have been prepared by methods substantially similar to those described herein in the examples and methods known to those of skill in the art.

[0411] Preparation of intermediates Method CA1 - Synthesis of tert-butyl (1-(6-aminopyridin-3-yl)piperidin-4-yl)(methyl)carbamate (CA1) [ka] Step 1. tert-Butyl methyl(1-(6-nitropyridin-3-yl)piperidin-4-yl)carbamate (CA1.2) A reaction vial was charged with 5-fluoro-2-nitropyridine (CA1.1) (0.66 g, 4.67 mmol), tert-butylmethyl(piperidin-4-yl)carbamate (1.00 g, 4.67 mmol), and N,N-diisopropylethylamine (8.1 mL, 46.66 mmol), and DMSO (10 mL). After stirring at 120 °C for 1.5 h, the reaction mixture was cooled to room temperature and poured into water (50 mL). The precipitate was collected by filtration and dried under vacuum to give the desired product (CA1.2) (1.6 g, 100%) as a yellow solid. m / z = 275.2 [M+H] + , 1 H NMR(400 MHz,DMSO):δ 8.32(d,J=2.8 Hz,1H),8.21-8.17(m,1H),7.54(dd,J=3.0,9.3 Hz,1H),4.28-4.20(m,3H),3.11(t,J=11.5 Hz,2H),2.71(s,3H),1.79-1.66(m,4H),1.46-1.43(m,9H).

[0412] Step 2. tert-Butyl (1-(6-aminopyridin-3-yl)piperidin-4-yl)(methyl)carbamate (CA1) A solution of tert-butyl methyl (1-(6-nitropyridin-3-yl)piperidin-4-yl)carbamate (CA1.2) (1.6 g, 4.76 mmol) in methanol (30 mL) containing Pd / C was hydrogenated at atmospheric pressure for 6 hours. The reaction mixture was filtered through Celite and washed with methanol. The filtrate was concentrated in vacuo to give the product (CA1) (1.25 g, 86%) as a pink solid. m / z=245.1 [M+H] + , 1H NMR(400 MHz,DMSO):δ 7.67(d,J=2.5 Hz,1H),7.21(dd,J=3.0,8.8 Hz,1H),6.44(d,J=8.8 Hz,1H),5.43(s,2H),3.89-3.88(m,1H),3.44(d,J=11.6 Hz,2H),2.75(s,3H),2.64(d,J=11.6 Hz,2H),1.89-1.76(m,2H),1.63(d,J=11.1 Hz,2H),1.46(s,9H).

[0413] Method PA1 - Preparation of 5-cyclopentyl-6-((dimethylamino)methyl)pyridin-2-amine (PA1) [ka] Step 1. Methyl 6-amino-3-bromopicolinate (PA1.1): To a solution of methyl 6-aminopicolinate (PA1.0) (500 g, 3289.4 mmol, 1.0 equiv.) in acetonitrile (12.5 L) was added N-bromosuccinamide (644 g, 3618.4 mmol, 1.1 equiv.) portionwise over 30 minutes at room temperature. After stirring for 30 minutes at room temperature, the reaction mixture was quenched with 10% NaSO solution in water (3.0 L) and concentrated to remove acetonitrile. The residue was diluted with 10% NaSO solution in water (20 L) and extracted with 50% ethyl acetate in hexane (10 L × 5). The combined organic layers were dried over sodium sulfate and concentrated under reduced pressure. The residue was triturated with 25% ethyl acetate in hexane (1 L) to give PA1.1. MS (ES): m / z 231-233 [M+2] + Note: The other regioisomer (methyl 6-amino-5-bromopicolinate) was also formed and isolated via silica purification. The desired regioisomer was confirmed by H NMR and NOE analysis.

[0414] Step 2. Methyl 3-bromo-6-(bis(tert-butoxycarbonyl)amino)picolinate (PA1.2) To a solution of methyl 6-amino-3-bromopicolinate (PA1.1) (1100 g, 4782.6 mmol, 1.0 equiv.) in THF (20 L) was added DMAP (116.7 g, 956.5 mmol, 0.2 equiv.) and Boc anhydride (2502 g, 11478.2 mmol, 2.4 equiv.). After stirring at 75 °C for 1.5 h, the reaction was concentrated, and the residue was diluted with brine solution (10 L) and extracted with ethyl acetate (2 x 10 L). The combined organic layers were dried over sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by column chromatography eluting with 5% ethyl acetate in hexane. The isolated material was triturated with hexane (4 L) to give PA1.2 (1700 g, 82.79%) as a white solid. MS (ES): m / z 431-433 [M+2] + .

[0415] Step 3. Methyl 6-(bis(tert-butoxycarbonyl)amino)-3-(furan-3-yl)picolinate (PA1.3) To a solution of 3-bromo-6-(bis(tert-butoxycarbonyl)amino)picolinate (PA1.2) (730 g, 1693.7 mmol, 1.0 equiv.), furanboronic acid (379 g, 3387.4 mmol, 2.0 equiv.), and tripotassium phosphate (1078.3 g, 5086.2 mmol, 3.0 equiv.) in 1,4-dioxane (5.85 L) and water (1.46 L) that had been degassed with a stream of nitrogen for 20 min, bis(triphenylphosphine)palladium(II) dichloride (59.5 g, 84.8 mmol, 0.05 equiv.) was added. After stirring at 120 °C for 15 min, the reaction was cooled to room temperature and the organic layer was collected. The organic layer was filtered through a Celite bed and concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with 6.0% to 10% ethyl acetate in hexanes. The isolated material was triturated with n-pentane (500 mL) to give PA1.3. MS (ES): m / z 418 [M +1] + .

[0416] Step 4. Methyl 6-(bis(tert-butoxycarbonyl)amino)-3-(tetrahydrofuran-3-yl)picolinate (PA1.4) To a solution of methyl 6-(bis(tert-butoxycarbonyl)amino)-3-(furan-3-yl)picolinate (PA1.3) (191 g, 456.9 mmol, 1.0 equiv.) in methanol (1140 ml) and THF (955 ml), ammonium formate (115.1 g, 182.5 mmol, 4.0 equiv.), acetic acid (133.7 ml, 0.7 V), and 20% wet palladium hydroxide on carbon (133.7 g, 1:0.7 w / w) were added. After stirring for 24 hours at room temperature under a hydrogen gas atmosphere, the reaction mixture was combined with six other batches of the same scale prepared in the same manner. The reaction mixture was filtered through a Celite bed, and the filtrate was concentrated under reduced pressure. The filtrate was neutralized with saturated sodium bicarbonate solution (10 L) and extracted with DCM (10 L × 3) to give PA1.4 (1251 g, 92.6%). MS(ES): m / z 423 [M+1] +

[0417] Step 5. tert-Butyl (6-(hydroxymethyl)-5-(tetrahydrofuran-3-yl)pyridin-2-yl)carbamate (PA1.5) A solution of methyl 6-(bis(tert-butoxycarbonyl)amino)-3-(THF-3-yl)picolinate (PA1.4) (250 g, 592.4 mmol, 1.0 equiv.) in ethanol (2500 mL) was treated portionwise with sodium borohydride (135 g, 355.4 mmol, 6.0 equiv.). After stirring at 60 °C for 2 h, the reaction mixture was concentrated under reduced pressure, diluted with water (10 L), and extracted with DCM (4 × 10 L). The combined organic layers were washed with brine (10 L), passed through Na SO , and concentrated under reduced pressure to give PA1.5, which was combined with four other batches of the same scale prepared in the same way. The product (640 g, 73.49%) as a colorless gummy liquid turned to a white solid at room temperature after 2 days. MS (ES): m / z 295.0 [M +1] + .

[0418] Steps 6 and 7. tert-Butyl (6-((dimethylamino)methyl)-5-(tetrahydrofuran-3-yl)pyridin-2-yl)carbamate (PA1.6) To a solution of tert-butyl (6-(hydroxymethyl)-5-(THF-3-yl)pyridin-2-yl)carbamate (PA1.5) (440 g, 149.6 mmol, 1.0 equiv) in DCM (6.5 L) at 0 °C, N,N-diisopropylethylamine (581.4 g, 448.9 mmol, 3.0 equiv) was added dropwise. After stirring for 20 min, mesyl chloride (257.04 g, 2244 mmol, 1.5 equiv) was added slowly at 0 °C. After stirring from 0 °C to room temperature for 2 h, the reaction mixture was quenched with DM water (1 L) and extracted with DCM (3 × 2 L). The combined organic layers were washed with brine (10 L), passed through Na SO , and concentrated under reduced pressure to give the mesylated intermediate. The product was combined with another batch (200 g scale) prepared in the same manner. As a pale yellow liquid (700 g, crude, 86.44%). MS(ES): m / z 373.35 [M +1] + .

[0419] To a solution of the mesylated intermediate (350 g, 940.0 mmol, 1.0 equiv.) in acetonitrile (3.5 L), N,N-diisopropylethylamine (529.23 g, 423.0 mmol, 4.5 equiv.) and dimethylamine hydrochloride (152.41 g, 1880.0 mmol, 2.0 equiv.) were added dropwise at room temperature. After stirring at 90 °C for 3 h, the reaction mixture was evaporated to remove acetonitrile. The residue was quenched with DM water (1500 ml) and extracted with DCM (3 × 3 L). The combined organic layers were washed with brine (10 L), dried over NaSO, and concentrated under reduced pressure to give PA1.6. The product was combined with another batch on the same scale prepared in the same manner. 700 g (quantitative yield) was obtained as a brown semi-solid. MS (ES): m / z 322.39 [M +1] + .

[0420] Step 8. 5-Cyclopentyl-6-((dimethylamino)methyl)pyridin-2-amine (PA1) To a solution of tert-butyl (6-((dimethylamino)methyl)-5-(THF-3-yl)pyridin-2-yl)carbamate (PA1.6) (700 g, 2180.7 mmol, 1.0 equiv) in DCM (5.0 L) at 0 °C was added trifluoroacetic acid (2.1 L, 3 volumes). After stirring at 70 °C for 2 h, the reaction mixture was evaporated. The residue was diluted with DM water (2 L) and extracted with heptane (2 × 1 L). The aqueous layer was neutralized with 10% sodium hydroxide solution and extracted with 15% methanol in DCM (4 × 3 L). The combined organic layers were dried over sodium sulfate and concentrated under reduced pressure. The residue was triturated with 20% ethyl acetate in hexanes (1 L) and diethyl ether (500 mL) to give PA1 as a light brown solid. (330 g, 68.47%) MS(ES): m / z 222.30 [M+1] + .

[0421] Method PA2 - Preparation of 6-((dimethylamino)methyl)-5-(tetrahydro-2H-pyran-4-yl)pyridin-2-amine (PA2) [ka] Step 1. Methyl 6-amino-3-bromopicolinate (PA2.1) To a solution of methyl 6-aminopicolinate (PA2.0) (500 g, 3289.4 mmol, 1.0 equiv.) in acetonitrile (12.5 L) was added N-bromosuccinimide (644 g, 3618.4 mmol, 1.1 equiv.) in portions over 30 min at room temperature. After stirring for 30 min at room temperature, the reaction mixture was quenched with 10% NaSO solution in water (3 L) and evaporated to remove acetonitrile. The residue was diluted with 10% NaSO solution in water (20 L) and extracted with 50% ethyl acetate in hexane (10 L × 5). The combined organic layers were dried over sodium sulfate and concentrated under reduced pressure. The residue was triturated with 25% ethyl acetate in hexane to give PA2.1 (methyl 6-amino-3-bromopicolinate). The product was combined with three other batches prepared in the same manner at a scale of 500 gm + 250 gm + 250 g = 1.5 kg total. (1300 g, 57.07%). MS(ES): m / z 231-233 [M+2] + ]. Note: The other regioisomer (methyl 6-amino-5-bromopicolinate) was also formed and was isolated via silica purification. The required regioisomer was confirmed by 1H NMR and NOE analysis.

[0422] Step 2. Methyl 3-bromo-6-(bis(tert-butoxycarbonyl)amino)picolinate (PA2.2) To a solution of methyl 6-amino-3-bromopicolinate (PA2.1) (1100 g, 4782.6 mmol, 1.0 equiv.) in THF (20 L) was added DMAP (116.7 g, 956.5 mmol, 0.2 equiv.) and Boc anhydride (2502 g, 11478.2 mmol, 2.4 equiv.). After stirring at 75 °C for 1.5 h, the reaction mixture was concentrated in vacuo, diluted with brine solution, and extracted with ethyl acetate (2 × 10 L). The combined organic layers were dried over sodium sulfate and concentrated in vacuo. The residue was purified by column chromatography eluting with 5% ethyl acetate in hexane. The isolated material was triturated with hexane (4 L) to give PA2.2 (1700 g, 82.79%) as a white solid. MS (ES): m / z 431-433 [M+2] + .

[0423] Step 3. tert-Butyl (5-bromo-6-(hydroxymethyl)pyridin-2-yl)carbamate (PA2.3) A solution of methyl 3-bromo-6-(bis(tert-butoxycarbonyl)amino)picolinate (PA2.2) (50 g, 116.27 mmol, 1.0 equiv) in ethanol (200 ml) was treated portionwise with sodium borohydride (26.3 g, 697.6 mmol, 6.0 equiv). After stirring at 70 °C for 2 h, the reaction mixture was concentrated in vacuo, quenched slowly with water (200 ml), and extracted into DCM (3 x 150 ml). The combined organic layers were washed with brine (100 ml), passed through a hydrophobic filter, and concentrated under reduced pressure to give PA2.3 (27 g, 79%) as a white solid. MS (ES): m / z 395 [M +1] +

[0424] Step 4, 5. tert-Butyl (5-bromo-6-((dimethylamino)methyl)pyridin-2-yl)carbamate (PA2.4) To a solution of tert-butyl (5-bromo-6-(hydroxymethyl)pyridin-2-yl)carbamate (PA2.3) (22.2 g, 73.2 mmol, 1.0 equiv.) and N-N-diisopropylethylamine (33.3 g, 256.3 mmol, 3.5 equiv.) in DCM (200 ml) was added methanesulfonyl chloride (12.5 g, 109.8 mmol, 1.5 equiv.) at 0° C. After stirring for 30 minutes, the reaction was quenched with water (100 ml) and extracted with ethyl acetate (3×40 ml). The combined organic layers were washed with brine, passed through a hydrophobic filter, and concentrated under reduced pressure to give the mesylate intermediate.

[0425] To the mesylate intermediate dissolved in acetonitrile (200 ml) was added dimethylamine (15 g, 183.0 mmol, 2.5 equiv.) and N-N-diisopropylethylamine (33.3 g, 256.3 mmol, 3.5 equiv.). After stirring at 70° C. for 1 hour, the reaction was quenched with water (100 ml) and extracted into ethyl acetate (3×40 ml). The combined organic layers were washed with brine, passed through a hydrophobic filter, and concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with 50% ethyl acetate / hexane to give (PA2.4) (17.0 g, 94.3%). MS (ES): m / z 330 [M +H] +

[0426] Step 6. tert-Butyl (5-(3,6-dihydro-2H-pyran-4-yl)-6-((dimethylamino)methyl)pyridin-2-yl)carbamate (PA2.5) To a solution of tert-butyl (5-bromo-6-((dimethylamino)methyl)pyridin-2-yl)carbamate (PA2.4) (50 g, 151.5 mmol, 1.0 equiv) in 1,4-dioxane:water (400 ml:100 ml) was added 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (47.7 g, 227.2 mmol, 1.5 equiv), and tripotassium phosphate (96.3 g, 454.5 mmol, 3.0 equiv). After degassing with N2 for 15 minutes, chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (11.9 g, 15.1 mmol, 0.1 equiv.) was added. After stirring at 140 °C for 4 hours, the reaction mixture was cooled to room temperature, diluted with water (1 L), and extracted with ethyl acetate (2 × 2 L). The combined organic extracts were washed with brine (1 L), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography eluting with 2.1% methanol in DCM to give PA2.5 (40 g, 79%). MS (ES): m / z 334.2 [M+H]. +

[0427] Step 7. tert-Butyl (6-((dimethylamino)methyl)-5-(tetrahydro-2H-pyran-4-yl)pyridin-2-yl)carbamate (PA2.6) To a suspension of palladium hydroxide (130 g) in methanol (600 ml) and THF (40 ml) was added tert-butyl (5-(3,6-dihydro-2H-pyran-4-yl)-6-((dimethylamino)methyl)pyridin-2-yl)carbamate (PA2.5) (130 g, 1.0 equiv.). Hydrogen gas was purged through the reaction mixture at room temperature for 4 hours. After completion of the reaction, the reaction mixture was filtered through a celite bed and washed with methanol. The filtrate was concentrated under reduced pressure to give PA2.6 (120 g, 91.75%). MS (ES): m / z 336.2 [M+H] +

[0428] Step 8. 6-((Dimethylamino)methyl)-5-(tetrahydro-2H-pyran-4-yl)pyridin-2-amine (PA2) To a solution of tert-butyl (6-((dimethylamino)methyl)-5-(tetrahydro-2H-pyran-4-yl)pyridin-2-yl)carbamate (PA2.6) (120 g, 356.9 mmol, 1.0 equiv) in DCM (1.2 L) was slowly added trifluoroacetic acid (360 ml). After stirring at 55° C. for 2 h, the reaction mixture was neutralized using saturated sodium hydroxide solution and extracted with 10% methanol in DCM (4×10 L). The combined organic layers were concentrated under reduced pressure to give PA2 (66 g, 78.40%). MS (ES): m / z 236.1 [M +H] +

[0429] Method PA3 - Preparation of 1-(6-amino-2-((dimethylamino)methyl)pyridin-3-yl)piperidin-4-ol (PA3) [ka] Step 1. 6-Chloro-3-(4-hydroxypiperidin-1-yl)picolinaldehyde (PA3.1) To a solution of 6-chloro-3-fluoropicolinaldehyde PA3.0 (1.0 g, 6.28 mmol, 1.0 equiv.) and piperidin-4-ol (1.0 g, 10.04 mmol, 1.6 equiv.) in N,N-dimethylformamide (10 ml) was added potassium carbonate (2.6 g, 18.84 mmol, 3.0 equiv.). After stirring at 100 °C for 1 h, the reaction mixture was cooled to room temperature, diluted with ice-cold water (100 ml), and extracted with ethyl acetate (3 × 40 ml). The combined organic extracts were washed with brine (80 ml), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography eluting with a 30% ethyl acetate in hexane gradient to give PA3.1 (1.2 g, 79.54%). MS (ES): m / z 241.07 [M+H]. +

[0430] Step 2. 1-(6-chloro-2-((dimethylamino)methyl)pyridin-3-yl)piperidin-4-ol (PA3.2) To a cooled solution of PA3.1 (1.2 g, 5.00 mmol, 1.0 equiv.) in 1,2-dichloroethane (20 ml) containing acetic acid (2.4 ml) at 0 °C, which had been purged with dimethylamine gas for 30 min, sodium triacetoxyborohydride (7.4 g, 35 mmol, 7.0 equiv.) was added portionwise. After stirring at room temperature for 16 h, the reaction mixture was diluted with ice-cold water (100 ml) and extracted with DCM (4 × 40 ml). The combined organic extracts were washed with brine (90 ml), dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography using a 3.0% methanol in DCM gradient elution to give PA3.2 (0.450 g, 33.46%). MS (ES): m / z 270.2 [M+H]. +

[0431] Step 3. N-(6-((dimethylamino)methyl)-5-(4-hydroxypiperidin-1-yl)pyridin-2-yl)cyclopropanecarboxamide (PA3.3) To a solution of 1-(6-chloro-2-((dimethylamino)methyl)pyridin-3-yl)piperidin-4-ol PA3.2 (0.45 g, 1.67 mmol, 1.0 equiv.) in 1,4-dioxane (10 mL), cyclopropylcarboxamide (0.156 g, 1.83 mmol, 1.1 equiv.) and potassium carbonate (0.691 g, 5.01 mmol, 3.0 equiv.) were added. After degassing with nitrogen gas for 10 minutes, 4,5-bis(diphenylphosphino)-9,9-dimethyloxane (0.19 g, 0.334 mmol, 0.2 equiv.) and tris(dibenzylideneacetone)dipalladium (0.152 g, 0.167 mmol, 0.1 equiv.) were added under a nitrogen atmosphere. After stirring at 120°C for 4 hours, the reaction mixture was diluted with water (100 ml) and extracted with ethyl acetate (100 ml x 3). The combined organic layers were washed with brine solution (50 ml), dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified using combi-flash silica eluting with 7% methanol / DCM to give PA3.3 (0.3 g, 66%). MS (ES): m / z 360.35 [M+H]+ .

[0432] Step 4. 1-(6-amino-2-((dimethylamino)methyl)pyridin-3-yl)piperidin-4-ol (PA3) To a solution of N-(6-((dimethylamino)methyl)-5-(4-hydroxypiperidin-1-yl)pyridin-2-yl)cyclopropanecarboxamide (0.5 g, 1.85 mmol, 1.0 equiv.) in methanol (7 ml) was added 5N sodium hydroxide solution (2.2 ml, 11.1 mmol, 6.0 equiv.). After stirring at 60° C. for 16 h, the reaction mixture was poured into ice-cold water (100 ml) and extracted with 10% chloroform / isopropyl alcohol (100 ml×3). The combined organic layers were dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by trituration with diethyl ether (20 ml×2) to give PA3 (0.3 g, 76.3%). MS (ES): m / z 251.35 [M+H]+

[0433] Method PA5 - Preparation of 2-(4-(6-aminopyridin-3-yl)morpholin-2-yl)propan-2-ol (PA5) [ka] Step 1. tert-Butyl 2-(2-hydroxypropan-2-yl)morpholine-4-carboxylate (PA5.1) To a solution of 4-(tert-butyl) 2-ethyl morpholine-2,4-dicarboxylate (PA5.0) (10 g, 38.61 mmol, 1.0 equiv.) in THF (100 ml) was added dropwise methylmagnesium bromide solution (3.0 M in diethyl ether) (100 ml) at 0 °C. After stirring at room temperature for 15 minutes, the reaction mixture was quenched with water (500 ml) and extracted with ethyl acetate (3 × 100 ml). The combined organic extracts were washed with brine (200 ml), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was used in the next step without further purification. PA5.1 (12.0 g, quant.%) MS (ES): m / z 245.3 [M + H] +

[0434] Step 2. 2-(morpholin-2-yl)propan-2-ol (PA5.2) To a solution of (PA5.1) (10.0 g, 40.81 mmol, 1.0 equiv.) in DCM (100 ml) was added trifluoroacetic acid (35 ml) dropwise at 0° C. The reaction mixture was stirred at room temperature for 30 minutes. After completion of the reaction, the reaction mixture was concentrated under reduced pressure and extracted with DCM (250 ml). The organic layer was concentrated under reduced pressure to give a solid material, which was used in the next step without further purification. PA5.2 (13 g, quant.%) MS (ES): m / z 146.1 [M +H] +

[0435] Step 3. 2-(4-(6-nitropyridin-3-yl)morpholin-2-yl)propan-2-ol (PA5.3) To a solution of 5-bromo-2-nitropyridine (7.0 g, 0.034 mol, 1.0 equiv.) in 1,4-dioxane (65.0 mL), 2-(morpholin-2-yl)propan-2-ol (PA5.2) (4.96 g, 0.034 mol, 1.0 equiv.) and potassium phosphate tripotassium (21.65 g, 0.102 mol, 3.0 equiv.) were added. After degassing with argon for 20 minutes, tris(dibenzylideneacetone)palladium(0) (3.1 g, 0.0034 mmol, 0.1 equiv.) and Xantphos (39.3 g, 0.068 mol, 0.2 equiv.) were added. After stirring at 120 °C for 3 hours, the reaction mixture was poured into water (250 mL) and extracted with ethyl acetate (300 mL x 3). The combined organic layers were washed with brine (200 ml), dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (0-40% ethyl acetate in hexanes gradient elution) to give PA5.3 (5.0 g, 38.80%). MS (ES): m / z 267.12 (M+H). + .

[0436] Step 4. 2-(4-(6-aminopyridin-3-yl)morpholin-2-yl)propan-2-ol (PA5) To a suspension of 10% Pd / C (2.2 g) in methanol (50 ml) was added a solution of 2-(4-(6-nitropyridin-3-yl)morpholin-2-yl)propan-2-ol (PA5.3) (4.0 g, 14.97 mmol, 1.0 equiv.) in methanol (10 ml) under a nitrogen atmosphere. After bubbling H2 (gas) through the reaction mixture for 3 hours, the reaction mixture was filtered through a bed of Celite and washed with methanol. The filtrate was concentrated under reduced pressure to give PA5 (2.2 g, 61.9%). MS (ES): m / z 237.18 [M+H] + ,

[0437] The following aniline intermediates in Table A1 were prepared according to any of the intermediate methods PA1-PA5 described above. [Table A1]

[0438] Method CB1—Synthesis of 9-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)-3-oxa-9-azaspiro[5.5]undecane (CB1) [ka] Step 1. 9-(4-Bromopyridin-2-yl)-3-oxa-9-azaspiro[5.5]undecane (CB1.2) A microwave vial was charged with 4-bromo-2-fluoropyridine (CB1.1) (300 mg, 1.7 mmol), 3-oxa-9-azaspiro[5.5]undecane (291 mg, 1.8 mmol), THF (3.0 mL), and triethylamine (862 mg, 8.5 mmol). After stirring in a microwave at 150 °C for 40 min, the reaction mixture was diluted with ethyl acetate (10 mL) and washed with water (10 mL). The aqueous layer was extracted with ethyl acetate (10 mL). The combined organic layers were dried (MgSO), filtered, and concentrated in vacuo. The residue was purified by silica chromatography (gradient elution: 0–50% ethyl acetate in cyclohexane) to afford (CB1.2) (317 mg, 60%) as a white solid. 1 H NMR(400 MHz,CDCl3)d 7.97(d,J=5.3 Hz,1H),6.79(s,1H),6.72(d,J=5.3 Hz,1H),3.70(t,J=5.3 Hz,4H),3.52(t,J=5.8 Hz,4H),1.63(t,J=5.5 Hz,4H),1.58-1.53(m,4H).

[0439] Step 2. 9-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)-3-oxa-9-azaspiro[5.5]undecane (CB1) A reaction vial was charged with 9-(4-bromopyridin-2-yl)-3-oxa-9-azaspiro[5.5]undecane (CB1.2) (317 mg, 1.0 mmol), bis(pinacolato)diboron (310 mg, 1.2 mmol), and potassium acetate (250 mg, 2.5 mmol), and dioxane (5 mL) was added. [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) was complexed with dichloromethane (42 mg, 0.05 mmol). After stirring overnight at 110 °C, the mixture was diluted with ethyl acetate (20 mL) and washed with water (20 mL). The combined organic layers were washed with brine (10 mL), passed through an Isolute® hydrophobic frit, and concentrated in vacuo. The residue was purified by silica chromatography (gradient elution 0-100% ethyl acetate in isohexane) to give (CB1) (143 mg, 40%) as a yellow oil. 1 H NMR(400 MHz,CDCl3)d 8.21(d,J=4.3 Hz,1H),7.05(s,1H),6.91(d,J=4.9 Hz,1H),3.70(t,J=5.1 Hz,4H),3.58-3.52(m,4H),1.67-1.61(m,4H),1.59-1.52(m,4H),1.34(s,12H).

[0440] Method CB2 - Synthesis of N-(4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (CB2) [ka] Step 1: N-(4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (CB2) To a solution of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoic acid (CB2.1) (518 mg, 2.0 mmol) and 4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (500 mg, 1.7 mmol) in DMF (4 mL) and DIPEA (910 mL, 5.22 mmol) was added HATU (992 mg, 2.61 mmol). After stirring overnight at room temperature, the mixture was diluted with ethyl acetate and washed successively with aqueous NaHCO3, water, and brine. The organic layer was collected, dried (Na2SO4), filtered, and evaporated to dryness to give crude (CB2) (900 mg, quantitative) as a yellow oil. The product was used in the next step without further purification. m / z = 518.2 [M+H] +

[0441] Method CB3 - Synthesis of 1-(piperidin-1-yl)-2-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)oxy)ethan-1-one (CB3) [ka] Step 1. 2-((4-Bromopyridin-2-yl)oxy)-1-(piperidin-1-yl)ethan-1-one (CB3.2) To a solution of 2-((4-bromopyridin-2-yl)oxy)acetic acid (CB3.1) (162 mg, 0.6 mmol) in DMF (5 mL) was added 1-hydroxybenzotriazole hydrate (120 mg, 0.8 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (170 mg, 0.8 mmol). After stirring at room temperature for 1 h, piperidine (81 mL, 0.8 mmol) was added. After stirring for 2 days, the mixture was diluted with diethyl ether, washed with water, passed through an Isolute® hydrophobic frit, and concentrated in vacuo to afford CB3.2 (74 mg, 36%) as a clear oil. 1H NMR(400 MHz,DMSO)d 8.07(d,J=5.6 Hz,1H),7.29-7.25(m,1H),7.24(s,1H),5.11(s,2H),3.45(s,4H),1.68-1.60(m,4H),1.48-1.44(m,2H).

[0442] Step 2. 1-(Piperidin-1-yl)-2-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)oxy)ethan-1-one (CB3) The reaction was carried out according to the procedure outlined in Method CB1, Step 2, using 2-((4-bromopyridin-2-yl)oxy)-1-(piperidin-1-yl)ethan-1-one (CB3.2) to afford the crude product (CB3) as a brown oil, which was used directly in the next step without purification.

[0443] Method CB4 - Synthesis of 4-bromo-2-(2,6-difluorophenyl)pyridine (CB4) [ka] Step 1. 4-Bromo-2-(2,6-difluorophenyl)pyridine (CB4) Under a nitrogen atmosphere, isopropylmagnesium chloride-lithium chloride complex (1.3 M in THF, 2.3 mL, 2.9 mmol) was added dropwise to a solution of 1-bromo-2,6-difluorobenzene (530 mg, 2.7 mmol) in THF (5 mL) at −78° C. After stirring for 1 h, zinc chloride (2 M in THF, 2.1 mL, 4.2 mmol) was added. After stirring at 0° C. for 30 min, a stirred suspension of 2,4-dibromopyridine (CB4.1) (500 mg, 2.1 mmol) and tetrakis(triphenylphosphine)palladium(0) (122 mg, 0.1 mmol) in THF (7 mL) was added. After stirring overnight at room temperature, the mixture was diluted with ethyl acetate and washed with water and brine. The organic phase was dried (NaSO), filtered, and concentrated in vacuo. The residue was purified by silica chromatography eluting with 25% ethyl acetate in cyclohexane to give CB4 (281 mg, 49%) as a colorless oil.1 H NMR(400 MHz,CDCl3)d 8.58(d,J=5.3 Hz,1H),7.70-7.67(m,1H),7.53-7.48(m,1H),7.43-7.33(m,1H),7.02(t,J=7.8 Hz,2H).

[0444] Method CB5 - Synthesis of 4-(4-bromopyridin-2-yl)tetrahydro-2H-pyran-4-carbonitrile (CB5) [ka] Step 1. 4-(4-Bromopyridin-2-yl)tetrahydro-2H-pyran-4-carbonitrile (CB5) To a solution of LDA (2 M in THF / heptane / ethylbenzene, 1.6 mL, 3.1 mmol) diluted with THF (10 mL) at −78 °C under a nitrogen atmosphere, 4-cyanotetrahydro-4H-pyran (340 mL, 3.1 mmol) was added dropwise. After stirring at −78 °C for 1 h, a solution of 2,4-dibromopyridine (CB4.1) (600 mg, 2.5 mmol) in THF (5 mL) was added dropwise. After warming to room temperature and stirring for an additional 4 h, the reaction was quenched with aqueous ammonium chloride solution and extracted with ethyl acetate (×2). The combined organic extracts were washed with brine, dried (NaSO), filtered, and evaporated under reduced pressure. The residue was purified by silica chromatography (gradient elution, 0–100% ethyl acetate in cyclohexane) to afford CB5 (649 mg, 95%) as a white solid. 1 H NMR(400 MHz,CDCl3)d 8.44(d,J=5.3 Hz,1H),7.63-7.61(m,1H),7.38(dd,J=1.5,5.3 Hz,1H),4.15-4.08(m,2H),3.64-3.55(m,2H),2.13-1.99(m,2H),1.95-1.86(m,2H).

[0445] The following boronate intermediates in Table A2 were prepared according to Method CB1 for the intermediates described above. [Table A2]

[0446] Method PB1 - Preparation of tert-butylmethyl(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)carbamate (PB1) [ka] Step 1. tert-Butyl (4-bromopyridin-2-yl)carbamate (PB1.1) To 4-bromopyridin-2-amine (PB1.0) (1.0 g, 5.376 mmol, 1.0 equiv.) dissolved in DCM was added DMAP (0.066 g, 0.5376, 0.1 equiv.) and triethylamine (1.64 g, 16.1 mmol, 3.0 equiv.). After stirring for 10 min, Boc anhydride (1.4 g, 6.45 L, 1.2 equiv.) was added. After stirring for 30 min, the reaction mixture was transferred to water (100 mL) and extracted with DCM (100 mL x 3). The combined organic layers were washed with brine solution (20 mL), dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel purification eluting with 2% DCM in methanol to give PB1.1 (1.5 g, 95.2%). MS (ES): m / z 274.23 [M+2]+.

[0447] Step 2. tert-Butyl (4-bromopyridin-2-yl)(methyl)carbamate (PB1.2) To a solution of PB1.1 (1.5 g, 5.49 mmol, 1.0 equiv.) in THF (10 ml) at 0 °C was added 60% sodium hydride (0.438 g, 18.25 mmol, 2.0 equiv.). After stirring at 0 °C for 30 min, methyl iodide (0.56 ml, 6.09 mmol, 1.2 equiv.) was added. After stirring at room temperature for 30 min, the reaction mixture was transferred to ice-cold water (100 ml) and extracted with ethyl acetate (100 ml × 3). The combined organic layers were dried over sodium sulfate and concentrated under reduced pressure. The residue was triturated (20 ml diethyl ether and 10 ml pentane) to give PB1.2 (1.5 g, 95.12). MS (ES): m / z 288.34 [M+H] +

[0448] Step 3. tert-Butyl methyl (4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)carbamate (PB1) To a solution of PB1.2 (1.5 g, 5.24 mmol, 1.0 equiv.) and bispinacoldiborane (2.66 g, 10.48 mmol, 2.0 equiv.) in N,N-dimethylformamide (10 mL) was added potassium acetate (1.69 g, 17.30 mmol, 3.3 equiv.) at room temperature. After degassing using argon gas for 20 minutes, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II), complex with DCM (0.427 g, 0.52 mmol, 0.1 equiv.) was added. After stirring at 90 °C for 6 hours, the reaction mixture was poured into water (100 mL) and extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography eluting with 15% ethyl acetate in hexane to give PB1 (0.350 g, 20.16%). MS (ES): m / z 335.18 [M+2] +

[0449] Method PB2 - Preparation of (2-(dimethylamino)pyridin-4-yl)boronic acid (PB2) [ka] Step 1. 2-(Dimethylamino)pyridin-4-yl)boronic acid (PB2) To a stirred solution of 4-bromo-N,N-dimethylpyridin-2-amine (PB2.0) (0.5 g, 2.48 mmol, 1.0 equiv.) in dry 1,4-dioxane (5 mL), bispinacolatodiborane (0.95 g, 3.73 mmol, 1.5 equiv.) and potassium acetate (0.730 g, 7.46 mmol, 3.0 equiv.) were added. After degassing under an argon atmosphere for 15 minutes, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.110 g, 0.12 mmol, 0.05 equiv.) and tricyclohexylphosphine (0.05 g, 0.20 mmol, 0.08 equiv.) were added. After degassing under an argon atmosphere for 10 minutes and stirring at 85 °C for 3 hours, the reaction mixture was concentrated under reduced pressure to give PB2, which was used directly in the next step without further purification. (0.400g, quantitative yield). MS(ES):m / z 167.08 [M+H] +

[0450] Method PB3 - Preparation of 3-(6-bromopyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (PB3) [ka] Step 1. 3-(6-Bromopyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (PB3) To a solution of 4,6-dibromopyrimidine (PB3.0) (0.25 g, 1.051 mmol, 1.0 equiv.) in dioxane (15 mL) was added 8-oxa-3-azabicyclo[3.2.1]octane (0.1432 g, 1.26 mmol, 1.2 equiv.) and triethylamine (0.23 g, 2.21 mmol, 2.0 equiv.). After stirring at room temperature for 1 h, the reaction mixture was diluted with water (50 mL) and extracted into ethyl acetate (50 mL x 3). The combined organic layers were washed with brine solution (50 mL), dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography eluting with 6-8% ethyl acetate in hexane to give PB3 (0.38 g, quantitative yield). MS (ES): m / z 272.17 [M+H] +

[0451] Method PB4 - Preparation of 4-chloro-2-(tetrahydro-2H-pyran-4-yl)pyrimidine (PB4) [ka] Step 1. Tetrahydro-2H-pyran-4-carboximidamide hydrochloride (PB4.1) A solution of tetrahydro-2H-pyran-4-carbonitrile (PB4.0) (3.0 g, 0.024 mol, 1.0 equiv.) in methanol was purged with HCl gas for 1 hour. After cooling in the refrigerator overnight, the reaction mixture was filtered and washed with methanol. The filtrate was concentrated under reduced pressure to give PB4.1 (3.5 g, 78.76%). MS (ES): m / z 164.07 [M+H] +

[0452] Step 2. 2-(Tetrahydro-2H-pyran-4-yl)pyrimidin-4-ol (PB4.2) To a solution of tetrahydro-2H-pyran-4-carboximidamide hydrochloride (PB4.1) (3.5 g, 0.027 mol, 1.0 equiv.) in N,N-dimethylmethanamide (35 mL) was added methyl (E)-3-methoxyacrylate (4.68 g, 0.032 mol, 1.2 equiv.) and potassium carbonate (7.5 g, 0.054 mol, 2.0 equiv.). After stirring at 120 °C for 2 h, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with brine solution (100 mL), dried over sodium sulfate, and concentrated under reduced pressure to give PB4.2 (2.5 g, 65.26%). MS (ES): m / z 180.09 [M+H] +

[0453] Step 3. 4-Chloro-2-(tetrahydro-2H-pyran-4-yl)pyrimidine (PB4) To a solution of 2-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-ol (PB4.2) (1.0 g, 5.61 mmol, 1.0 equiv.) was added phosphorus oxychloride (3.2 ml, 27.7 mmol, 5.0 equiv.). After stirring at 100° C. for 1 h, the reaction mixture was neutralized with aqueous sodium bicarbonate (100 ml) and extracted with ethyl acetate (3×100 ml). The combined organic layers were washed with brine solution (100 ml), dried over sodium sulfate, and concentrated under reduced pressure to give PB4 (0.8 g, 72.57%). MS (ES): m / z 198.06 [M+H] +

[0454] Method PB5 - Preparation of 4-chloro-2-(cyclohexylethynyl)pyridine (PB5) [ka] Step 1. 4-Chloro-2-(cyclohexylethynyl)pyridine (PB5) To a solution of 2-bromo-4-chloropyridine (PB5.0) (2.48 g, 12.9 mmol, 1.5 equiv.), ethynylcyclohexane (1.0 g, 8.62 mmol, 1.0 equiv.), and N,N'-dimethylethylenediamine (0.3 g, 3.4 mmol, 0.4 equiv.) in toluene was added cesium carbonate (5.6 g, 17.24 mmol, 2.0 equiv.) and ferric chloride (0.2 g, 1.29 mmol, 0.15 equiv.). After stirring at 135 °C for 16 h, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 3). The combined organic layers were washed with brine solution (100 mL), dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography eluting with 2% ethyl acetate in hexane to give PB5 (1.2 g, 59%) MS(ES): m / z 219.08 [M+H] +

[0455] Method PB7 - Preparation of 4-bromo-2-(tetrahydro-2H-pyran-2-yl)pyridine (PB7) [ka] Step 1. 2-(3,4-Dihydro-2H-pyran-6-yl)-4-nitropyridine (PB7.1) To a stirred solution of 2-bromo-4-nitropyridine (PB7.0) (1.0 g, 4.92 mmol, 1.0 equiv.) in 1,4-dioxane (8 mL) and water (2 mL) was added 2-(3,4-dihydro-2H-pyran-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.2 g, 5.9 mmol, 1.2 equiv.) and cesium carbonate (3.2 g, 9.8 mmol, 2.0 equiv.). After degassing under argon for 15 minutes, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.40 g, 0.49 mmol, 0.1 equiv.) was added. After degassing under argon atmosphere for 10 minutes and stirring at 100°C for 1 hour, the reaction mixture was quenched with water (100ml) and extracted with ethyl acetate (100ml x 3). The combined organic layers were washed with brine solution, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography eluting with 10% ethyl acetate in hexane to give PB7.1 (0.62g, 61.04%). MS(ES): m / z 207.24 [M+H] +

[0456] Step 2. 2-(Tetrahydro-2H-pyran-2-yl)pyridin-4-amine (PB7.2) To a suspension of 10% palladium on carbon containing 50% water (0.3 g) in methanol (2 mL) was added a solution of PB7.1 (0.62 g, 3.0 mmol, 1.0 equiv) in methanol (3 mL) under a nitrogen atmosphere. Hydrogen gas was bubbled through the reaction mixture for 4 hours, after which the reaction mixture was filtered through Celite and washed with methanol. The filtrate was concentrated under reduced pressure to give PB7.2 (0.5 g, 93.30%). MS (ES): m / z 179.20 [M+H] +

[0457] Step 3. 4-Bromo-2-(tetrahydro-2H-pyran-2-yl)pyridine (PB7) To a stirred solution of PB7.2 (0.2 g, 1.12 mmol, 1.0 equiv) in 33% HBr in acetic acid (3 mL) was added aqueous NaNO (0.309 g, 4.48 mmol, 4.0 equiv) dropwise at 0 °C. After stirring at 0 °C for 15 min, Cu(I)Br (0.161 g, 1.12 mmol, 1.0 equiv) was added portionwise. After stirring at 0 °C for 30 min, the reaction mixture was neutralized to approximately pH 8 with saturated sodium bicarbonate solution and extracted with ethyl acetate (50 ml × 3). The combined organic layers were washed with brine solution (50 ml), dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography eluting with 2% methanol in DCM to give PB7 (0.15 g, 55.21%). MS (ES): m / z 243.16 [M+H] + .

[0458] Method P13 - Preparation of 4-bromo-6-(tetrahydro-2H-pyran-4-yl)pyrimidine (PB13) [ka] Step 1. 6-(Tetrahydro-2H-pyran-4-yl)pyrimidin-4-ol (PB13.1) To a solution of ethyl 3-oxo-3-(tetrahydro-2H-pyran-4-yl)propanoate (PB13.0) (1.0 g, 5.0 mmol, 1.0 equiv.) and formimidamide hydrochloride (0.6 g, 6.0 mmol, 1.2 equiv.) in methanol was added sodium methoxide (30% methanol) (1.34 g, 25.0 mmol, 5.0 equiv.). After stirring at room temperature for 16 hours, the reaction mixture was diluted with water (100 ml) and extracted with ethyl acetate (100 ml x 3). The combined organic layers were washed with brine solution (100 ml), dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography eluting with 2% methanol in DCM to give PB13.1 (0.85 g, 94.5%). MS (ES): m / z 180.09 [M+H] + .

[0459] Step 2. 4-Chloro-6-(tetrahydro-2H-pyran-4-yl)pyrimidine (PB13) To a solution of tetrahydro-2H-pyran-4-yl)pyrimidin-4-ol (PB13.1) (0.8 g, 4.4 mmol, 1.0 equiv.) was added phosphorus oxychloride (8 ml). After stirring at 90° C. for 1 hour, the reaction mixture was quenched with saturated sodium bicarbonate (50 ml) and water (50 ml) and extracted with ethyl acetate (100 ml×3). The combined organic layers were washed with brine solution (100 ml), dried over sodium sulfate, and concentrated under reduced pressure to give PB13 (0.75 g, 85.05%). MS (ES): m / z 198.09 [M+H] + .

[0460] Method P14 - Preparation of 3-(4-bromopyridin-3-yl)-8-oxa-3-azabicyclo[3.2.1]octane (PB14) [ka] Step 1. 3-(4-Bromopyridin-3-yl)-8-oxa-3-azabicyclo[3.2.1]octane (PB14) To a solution of PB14.0 (0.50 g, 1.76 mmol, 1.0 equiv.) and PB14.1 (0.339 g, 1.76 mmol, 1.1 equiv.) in toluene (5 mL) was added sodium tert-butoxide (0.304 g, 3.1 mmol, 1.8 equiv.). After degassing under argon for 20 minutes, tris(dibenzylideneacetone)dipalladium(0) (0.048 g, 0.05 mmol, 0.03 equiv.) and 4,5-bis(diphenylphosphino)-9,9-dimethylxane (0.111 g, 0.19 mmol, 0.1 equiv.) were added. After stirring at 110 °C for 16 hours, the reaction mixture was transferred to water (50 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic layers were washed with brine solution (20 mL), dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography eluting with 15-20% ethyl acetate in hexane to give PB14 (0.250 g, 53.07%) MS(ES): m / z 269.7 [M+H] + .

[0461] Method P15 - Preparation of 4-chloro-2-(4,5-dihydrofuran-3-yl)pyridine (PB15) [ka] Step 1. 4-Chloro-2-(4,5-dihydrofuran-3-yl)pyridine (PB15) To a solution of 2-bromo-4-chloropyridine (PB15.0) (0.5 g, 2.6 mmol, 1.0 equiv.) and 2-(4,5-dihydrofuran-3-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (PB15.1) (0.61 g, 3.1 mmol, 1.2 equiv.) in dioxane was added cesium carbonate (0.3 g, 1.04 mmol, 2.0 equiv.). After degassing at room temperature under argon for 15 minutes, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.19 g, 0.26 mmol, 0.1 equiv.) was added. After stirring at 110 °C for 2 hours, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic layers were washed with brine solution (50 mL), dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified using column chromatography eluting with 20% ethyl acetate in hexane to give PB15 (0.35 g, 74.17%) MS(ES): m / z 181.03 [M+H] + .

[0462] Method PB16 - Preparation of 4-iodo-1-methyl-5-phenyl-1H-imidazole (PB16) [ka] Step 1. 4-Iodo-1-methyl-5-phenyl-1H-imidazole (PB16) To a solution of 1-methyl-5-phenyl-1H-imidazole (PB16.0) (0.25 g, 1.58 mmol, 1.0 equiv.) in DCM, N-iodosuccinimide (0.7 g, 3.1 mmol, 2.0 equiv.) and trifluoroacetic acid (0.1 ml) were added. After stirring at room temperature for 16 hours, the reaction mixture was neutralized with aqueous sodium bicarbonate (50 ml) and extracted with ethyl acetate (50 ml x 3). The combined organic layers were washed with brine solution (50 ml), dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified using column chromatography eluting with 1% methanol in DCM to give PB16 (0.32 g, 71.2%). MS (ES): m / z 283.9 [M+H] + .

[0463] Method PB17 - Preparation of 5-bromo-1-methyl-2-phenyl-1H-imidazole (PB17) [ka] Step 1. 1-Methyl-2-phenyl-1H-imidazole (PB17.1) To a solution of 2-phenyl-1H-imidazole (PB17.0) (1.0 g, 6.94 mmol, 1.0 equiv.) in acetone (25 ml), potassium hydroxide (1.1 g, 20.82 mmol, 3.0 equiv.) was added. After stirring at room temperature for 10 minutes, methyl iodide (1.5 g, 10.41 mmol, 1.5 equiv.) was added. After completion of the reaction, the reaction mixture was diluted with water (100 ml) and extracted with ethyl acetate (100 ml x 3). The combined organic layers were washed with brine solution (100 ml), dried over sodium sulfate, and concentrated under reduced pressure to give PB17.1 (0.92 g, 84.3%). MS (ES): m / z 158.09 [M+H] + .

[0464] Step 2. 5-Bromo-1-methyl-2-phenyl-1H-imidazole (PB17) To a solution of 1-methyl-2-phenyl-1H-imidazole (PB17.1) (0.8 g, 5.05 mmol, 1.0 equiv) in isopropyl acetate (20 ml) was added potassium carbonate (0.13 mg, 1.01 mmol, 0.2 equiv). After stirring at room temperature for 10 minutes, N-bromosuccinimide (0.85 g, 4.81 mmol, 0.95 equiv) was added portionwise. After stirring at room temperature for 3 hours, the reaction mixture was diluted with water (100 ml) and extracted with ethyl acetate (100 ml x 3). The combined organic layers were washed with brine solution (100 ml), dried over sodium sulfate, and concentrated under reduced pressure to give PB17 (0.6 g, 50%). MS (ES): m / z 237.99 [M+H] + .

[0465] Method PB19 - Preparation of 4-chloro-5-ethylpyrimidin-2-amine (PB19) [ka] Step 1. 4-Chloro-5-ethylpyrimidin-2-amine (PB19) To a solution of 2-amino-5-ethylpyrimidin-4(3H)-one (PB19.0) (1.0 g, 7.19 mmol, 1.0 equiv.) in chloroform (10 ml) with 3–5 drops of N,N-dimethylformamide at room temperature was added phosphorus oxychloride (1.5 ml). After stirring at 70 °C for 6 h, the reaction mixture was quenched with saturated sodium bicarbonate solution, diluted with water (100 ml), and extracted with ethyl acetate (100 ml × 3). The combined organic layers were washed with brine solution (100 ml), dried over sodium sulfate, and concentrated under reduced pressure to give PB19 (0.9 g, 79.47%). MS (ES): m / z 159.04 [M+H] + .

[0466] Method PB20 - Preparation of 3-(5-chloropyrimidin-2-yl)-8-oxa-3-azabicyclo[3.2.1]octane (PB20) [ka] Step 1. 2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)pyrimidin-5-ol (PB20.1) To a solution of 2-(methylthio)-1,6-dihydropyrimidin-5-ol (PB20.0) (0.4 g, 2.81 mmol, 1.0 equiv.) in diglyme (6 mL) was added 8-oxa-3-azabicyclo[3.2.1]octane (0.36 g, 3.36 mmol, 1.15 equiv.). After stirring at 150 °C for 18 h, the reaction mixture was concentrated under reduced pressure to give PB13 (0.300 g, 52.18%). MS (ES): m / z 208.23 [M+H] + .

[0467] Step 2. 3-(5-chloropyrimidin-2-yl)-8-oxa-3-azabicyclo[3.2.1]octane (PB20) To a solution of 2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)pyrimidin-5-ol (PB20.1) (0.8 g, 3.86 mmol, 1.0 equiv.) was added phosphorus oxychloride (8 ml). After stirring at 90° C. for 1 hour, the reaction mixture was quenched with saturated sodium bicarbonate (50 ml) and water (50 ml) and extracted with ethyl acetate (100 ml×3). The combined organic layers were washed with brine solution (100 ml), dried over sodium sulfate, and concentrated under reduced pressure to give PB20 (0.70 g, 80.35%). MS (ES): m / z 226.6 [M+H] + .

[0468] The following aniline intermediates in Table A3 were prepared according to any of the intermediate methods PB1-PB20 described above. [Table A3]

[0469] Example 1. Method A Synthesis of 7-[[5-(4-methylpiperazin-1-yl)-2-pyridyl]amino]-4-(1H-pyrrol-3-yl)-2,3-dihydropyrrolo[3,4-c]pyridin-1-one (I-23) [ka] Step 1. 5-Bromo-2-chloro-N-methoxy-N-methylnicotinamide (1.2) A mixture of 5-bromo-2-chloronicotinic acid (1.1) (4.00 g, 16.92 mmol) in thionyl chloride (20 mL) was heated at 80 °C for 2 h. The mixture was concentrated in vacuo and azeotroped with toluene (2 × 10 mL). The residue was extracted with DCM and cooled to 0 °C. The mixture was treated with N,O-dimethylhydroxylamine hydrochloride (2.06 g, 21.15 mmol) followed by trimethylamine (7.1 mL, 50.75 mmol). After stirring for 1 h, the mixture was diluted with DCM, washed with 10% citric acid solution, saturated aqueous NaHCO solution, water, and brine, dried over NaSO, and concentrated in vacuo to give 1.2 (3.81 g, 81%) as a pale orange solid. m / z = 280.9 [M+H] + , 1 H NMR (400 MHz, CDCl3): δ 8.50(d,J=2.0 Hz,1 H),7.83-7.78(m,1 H),3.53(s,3 H),3.39(s,3 H).

[0470] Step 2. 5-Bromo-2-chloronicotinaldehyde (1.3) To a solution of 5-bromo-2-chloro-N-methoxy-N-methylnicotinamide (1.2) (3.80 g, 13.59 mmol) in dry THF (30 mL) at -10 °C under a nitrogen atmosphere, LiAlH (1 M solution in THF, 5.4 mL, 5.44 mmol) was added dropwise. After warming to room temperature and stirring for 18 h, the reaction was cooled to 0 °C, carefully quenched by the addition of 1 M KHSO solution, and extracted into EtOAc (3 × 20 mL). The combined extracts were washed with brine, dried over NaSO, and concentrated in vacuo. The residue was purified by silica gel chromatography (0–25% gradient elution with EtOAc in iso-hexane) to give 1.3 (2.31 g, 77%) as a white solid. m / z = 220.9 [M+H] + , 1H NMR (400 MHz, CDCl3): δ 10.37(s,1 H),8.66(d,J=2.6 Hz,1 H),8.33(d,J=2.6 Hz,1 H).

[0471] Step 3. 5-Bromo-2-chloro-3-(dimethoxymethyl)pyridine (1.4) To a solution of 5-bromo-2-chloronicotinaldehyde (1.3) (18.66 g, 84.6 mmol) in MeOH (50 mL) was added p-toluenesulfonic acid (1.61 g, 0.85 mmol) and trimethyl orthoformate (37 mL, 338.4 mmol). After stirring at reflux for 2 h, the solvent was removed in vacuo and the residue was extracted with EtOAc (100 mL). The organic solution was washed with saturated aqueous NaHCO (2 × 100 mL), water (100 mL), and brine (100 mL), dried over MgSO, filtered, and concentrated in vacuo to give 1.4 (16 g, 70%) as a clear oil. m / z = 267.5 [M+H] + , 1 H NMR(400 MHz,CDCl3)δ 8.42(d,1 H),8.08(d,1 H),5.53(s,1 H),3.41(s,6 H).

[0472] Step 4. Ethyl 5-bromo-2-chloro-3-(dimethoxymethyl)isonicotinate (1.5) To a degassed solution of LDA (2 M in THF / heptane / ethylbenzene, 5.16 mL, 10.32 mmol) in dry THF (25 mL) at −78 °C was added dropwise 5-bromo-2-chloro-3-(dimethoxymethyl)pyridine (1.4) (2.5 g, 9.38 mmol) in dry, degassed THF (10 mL). After 30 min, ethyl chloroformate (2.68 mL, 28.14 mmol) was added dropwise. After stirring at −50 °C for 40 min, the reaction was quenched with saturated aqueous NaHCO (25 mL) and allowed to warm to room temperature. The mixture was extracted with ethyl acetate (2 × 25 mL), and the combined organic phase was washed with brine (20 mL), dried over MgSO, filtered, and concentrated in vacuo. The residue was purified by column chromatography (0-50% gradient elution EtOAc in iso-hexane) to give 1.5 (2.01 g, 63%) as a yellow oil. m / z=339.7 [M+H] + , 1 H NMR(400 MHz,CDCl3)δ 8.52(s,1 H),5.53(s,1 H),4.43(q,2 H),3.41(s,6 H),1.41(t,3 H)

[0473] Step 5. Ethyl 5-bromo-2-chloro-3-formyl isonicotinate (1.6) To a solution of ethyl 5-bromo-2-chloro-3-(dimethoxymethyl)isonicotinate (1.5) (6.9 g, 20.4 mmol) in MeCN (200 mL) and HO (4 mL) was added lithium tetrafluoroborate (1 M in MeCN, 20.4 mL, 20.4 mmol). After stirring at 90 °C for 16 h, the reaction mixture was diluted with DCM and washed with HO (2 × 30 mL). The solvent was removed in vacuo to give 1.6 (5.8 g, 97%) as an orange oil. m / z = 293.6 [M+H] + , 1 H NMR(400 MHz,CDCl3)δ 10.35(s,1 H),8.73(s,1 H),4.53(q,2 H),3.41(s,6 H),1.44(t,3 H).

[0474] Step 6. 7-Bromo-4-chloro-2-(2,4-dimethoxybenzyl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (1.7) To a solution of ethyl 5-bromo-2-chloro-3-formyl isonicotinate (1.6) (5.8 g, 19.83 mmol) in DCM (40 mL) was added acetic acid (3.4 mL, 59.48 mmol). After stirring at room temperature for several minutes, 2,4-dimethoxybenzylamine (3.28 mL, 21.81 mmol) was added dropwise. After 3 h, sodium cyanoborohydride (1.87 g, 29.74 mmol) was added portionwise. After stirring at room temperature for 1 h, the mixture was filtered through a pad of Celite and the solvent was removed in vacuo. The residue was purified by column chromatography (0-60% gradient elution with EtOAc in iso-hexane) to give 1.7 (3.5 g, 44%) as a yellow oil. m / z = 366.5 [M+H] + , 1 H NMR(400 MHz,CDCl3)δ 8.53(s,1 H),7.28(d,J=7.7 Hz,1 H),6.49-6.42(m,2 H),4.75(s,2 H),4.28(s,2 H),3.86(s,3 H),3.80(s,3 H).

[0475] Step 7. 4-Chloro-2-(2,4-dimethoxybenzyl)-7-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (1.8) To a solution of 7-bromo-4-chloro-2-(2,4-dimethoxybenzyl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (1.7) (2 g, 5.03 mmol) in dry 1,4-dioxane (60 mL) was added CsCO (3.27 g, 10.06 mmol) and 5-(4-methylpiperazin-1-yl)pyridin-2-amine (1.06 g, 5.54 mmol). After purging with a stream of N for 10 min, Xantphos (350 mg, 0.604 mmol) and Pd(dba) (461 mg, 0.503 mmol) were added and the mixture was again degassed for 10 min. After stirring at 120 °C for 2 h, the reaction mixture was cooled to room temperature, poured into saturated aqueous NH4Cl solution (30 mL), filtered through a Celite pad, and extracted with ethyl acetate (2 x 25 mL). The combined organic phases were washed with brine (25 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (0-100% gradient elution EtOAc in iso-hexane) to give 1.8 (1.6 g, 62%) as a dark orange solid. m / z = 478.1 [M+H] + , 1 H NMR(400 MHz,DMSO):δ 9.41-9.38(m,1 H),8.53(d,J=9.1 Hz,1 H),8.05-8.02(m,1 H),7.52-7.46(m,1 H),7.08(d,J=8.8 Hz,1 H),4.75(s,2 H),3.19-3.13(m,4 H),2.53-2.47(m,4 H),2.28(s,3 H),1.59(s,9 H).

[0476] Step 8. tert-Butyl 3-(2-(2,4-dimethoxybenzyl)-7-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl)-1H-pyrrole-1-carboxylate (1.9) To a mixture of 4-chloro-2-(2,4-dimethoxybenzyl)-7-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (1.8) (300 mg, 0.58 mmol) and tert-butyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrole-1-carboxylate (190 mg, 0.64 mmol) in 1,4-dioxane (10 mL) and water (1 mL) was added Pd(dppf)Cl 1:1 DCM complex (48 mg, 10 mol%) and CsCO (480 mg, 1.47 mmol). After degassing with nitrogen and stirring at 100 °C for 6 h, the reaction mixture was diluted with EtOAc (20 mL). The combined organic solution was washed with water (15 mL) and brine (15 mL), dried over NaSO, and concentrated in vacuo. The residue was purified by silica gel chromatography (gradient elution: 0-10% MeOH in DCM) to afford 1.9 (270 mg, 71%) as an orange residue, which was used directly in the next step.

[0477] Step 9. 7-[[5-(4-methylpiperazin-1-yl)-2-pyridyl]amino]-4-(1H-pyrrol-3-yl)-2,3-dihydropyrrolo[3,4-c]pyridin-1-one (I-23) A 5 mL microwave vial was charged with tert-butyl 3-(2-(2,4-dimethoxybenzyl)-7-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl)-1H-pyrrole-1-carboxylate (1.9) (270 mg, 0.42 mmol) and TFA (mL). After stirring in a microwave at 160° C. for 10 minutes, the mixture was concentrated in vacuo and the residue was purified by preparative HPLC to give the title compound (I-23) (12 mg, 7%) as an orange solid. m / z=390 [M+H] + , 1H NMR(400 MHz,DMSO):δ 11.16(s,1 H),9.68(s,1 H),9.20(s,1 H),8.06(d,J=2.3 Hz,1 H),7.51(dd,J=2.7,9.0 Hz,1 H),7.29-7.26(m,1 H),7.04(d,J=8.8 Hz,1 H),6.91(d,J=1.5 Hz,1 H),6.66(s,1 H),4.63(s,2 H),3.54-3.48(m,5 H),3.30-3.28(m,2 H),3.07-3.02(m,3 H),2.64(s,2 H).

[0478] Example 2. Method B. Synthesis of 4-cyclopropyl-7-((5-(piperazin-1-yl)pyridin-2-yl)amino)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (I-12) [ka] Step 1. 2-Bromo-5-chloronicotinaldehyde (2.2) A solution of 2,3-dibromo-5-chloropyridine (2.1) (25.29 g, 93.21 mmol) in THF (200 mL) at −40° C. under a nitrogen atmosphere was treated dropwise with isopropylmagnesium chloride (2 M in THF, 50.8 mL, 101.59 mmol). After stirring for 1 h, DMF (21.1 mL, 272.20 mmol) was added dropwise. After warming to room temperature over 30 min, the reaction was quenched with 1 M HCl and extracted into tert-butyl methyl ether (3×). The combined extracts were washed with saturated aqueous NaHCO solution, water, and brine, dried over NaSO, filtered, and concentrated in vacuo to give 2.2 (20.35 g, 99%) as a beige solid. m / z=221.2 [M+H] + , 1 H NMR (400 MHz, CDCl3): δ 10.29(s,1 H),8.54(d,J=2.8 Hz,1 H),8.13(d,J=2.8 Hz,1 H).

[0479] Step 2. 2-Bromo-5-chloronicotinaldehyde (2.3) To a solution of 2-bromo-5-chloronicotinaldehyde (2.2) (25.40 g, 115.22 mmol) in methanol (300 mL) was added triethyl orthoformate (37.8 mL, 345.66 mmol) and p-toluenesulfonic acid monohydrate (2.19 g, 11.52 mmol). After stirring at reflux for 18 h, the cooled mixture was concentrated in vacuo and passed through a silica pad eluting with 20% EtOAc in iso-hexane to give 2.3 (28.61 g, 93%) as a yellow oil. m / z=267.2 [M+H] + , 1 H NMR (400 MHz, CDCl3): δ 8.32(d,J=2.5 Hz,1 H),7.89(d,J=2.5 Hz,1 H),5.47(s,1 H),3.41(s,6 H).

[0480] Step 3. Ethyl 2-bromo-5-chloro-3-(dimethoxymethyl)isonicotinate (2.4) To a solution of LDA (2 M in THF / heptane / ethylbenzene, 69.8 mL, 139.5 mmol) in dry THF (130 mL) at −50° C. under a nitrogen atmosphere, a solution of 2-bromo-5-chloro-3-(dimethoxymethyl)pyridine (2.3) (28.60 g, 107.31 mmol) in dry THF (70 mL) was added dropwise over 40 min. After the addition, the mixture was stirred for an additional 40 min, after which ethyl chloroformate (30.7 mL, 321.93 mmol) was added dropwise. After stirring for 40 min at −50° C., the reaction was quenched with saturated aqueous NaHCO solution and extracted with EtOAc (3×). The combined extracts were washed with brine, dried over NaSO, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography (0-30% gradient elution EtOAc in iso-hexane) to give 2.4 (26.51 g, 73%) as a pale yellow oil. m / z=339.6 [M+H] + , 1H NMR (400 MHz, CDCl3): δ 8.38(s,1 H),5.50-5.49(m,1 H),4.42(q,J=7.2 Hz,2 H),3.43-3.42(m,6 H),1.40(dd,J=7.2,7.2 Hz,3 H).

[0481] Step 4. Ethyl 2-bromo-5-chloro-3-formyl isonicotinate (2.5) A mixture of ethyl 2-bromo-5-chloro-3-(dimethoxymethyl)isonicotinate (2.4) (26.50 g, 78.27 mmol) and lithium tetrafluoroborate (10.27 g, 109.57 mmol) in acetonitrile (250 mL) and water (15 mL). After stirring at 75 °C for 18 h, the cooled mixture was concentrated in vacuo, extracted with EtOAc, washed with saturated aqueous NaHCO solution, water, and brine, dried over NaSO, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography (0-30% gradient elution with EtOAc in iso-hexane) to give 2.5 (13.94 g, 61%) as a yellow oil. m / z = 293.5 [M+H] + , 1 H NMR (400 MHz, DMSO): δ 8.62(s,1 H),7.21(d,J=8.3 Hz,1 H),4.68(s,2 H),3.87(s,3 H).

[0482] Step 5. 4-Bromo-7-chloro-2-(2,4-dimethoxybenzyl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (2.6) To a solution of ethyl 2-bromo-5-chloro-3-formyl isonicotinate (2.5) (13.93 g, 47.62 mmol) and acetic acid (8.18 mL, 142.87 mmol) in DCM (200 mL) was added MgSO4 and 2,4-dimethoxybenzylamine (7.87 mL, 52.38 mmol). After stirring at room temperature for 18 h, sodium borohydride (2.70 g, 71.43 mmol) was added portionwise. After stirring for 2 h, the mixture was diluted with DCM, washed with saturated aqueous NaHCO3 solution, water, and brine, dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography (0-50% gradient elution with EtOAc in DCM). The resulting yellow residue was triturated with diethyl ether to give 2.6 (7.12 g, 38%) as a white solid. m / z = 398.6 [M+H] + , 1 H NMR(400 MHz,DMSO):δ 8.61(s,1 H),7.20(d,J=8.3 Hz,1 H),6.64(d,J=2.3 Hz,1 H),6.54(dd,J=2.1,8.2 Hz,1 H),4.67(s,2 H),4.36(s,2 H),3.87(s,3 H),3.81(s,3 H).

[0483] Step 6. 7-Chloro-4-cyclopropyl-2-(2,4-dimethoxybenzyl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (2.7) To a mixture of 4-bromo-7-chloro-2-(2,4-dimethoxybenzyl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (2.6) (200 mg, 0.503 mmol) and Pd(PPh) (58 mg, 0.05 mmol) in dry, nitrogen-degassed THF (8 mL) was added cyclopropylzinc bromide (0.5 M in THF, 2.02 mL, 1.01 mmol). After stirring at room temperature for 18 h, the reaction was quenched with NH4Cl solution and extracted into EtOAc (×2). The combined extracts were washed with brine, dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography (0–50% gradient elution with EtOAc in iso-hexane) to give 2.7 (26.51 g, 73%) as a yellow residue. m / z = 359.6 [M+H] + , 1 H NMR(400 MHz,DMSO):δ 8.54(s,1 H),7.16(d,J=8.3 Hz,1 H),6.65(d,J=2.0 Hz,1 H),6.55(dd,J=2.0,8.3 Hz,1 H),4.68(s,2 H),4.55(s,2 H),3.87(s,3 H),3.81(s,3 H),2.16-2.04(m,1 H),1.07(d,J=6.3 Hz,4 H).

[0484] Step 7. tert-Butyl 4-(6-((4-cyclopropyl-2-(2,4-dimethoxybenzyl)-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-7-yl)amino)pyridin-3-yl)piperazine-1-carboxylate (2.8) The reaction was carried out according to the procedure outlined in Example 1. Step 7 of Method A using 7-chloro-4-cyclopropyl-2-(2,4-dimethoxybenzyl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (2.7) and tert-butyl 4-(6-aminopyridin-3-yl)piperazine-1-carboxylate gave 2.8 as a yellow residue, which was used directly in the next step.

[0485] Step 8. 4-Cyclopropyl-7-((5-(piperazin-1-yl)pyridin-2-yl)amino)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (I-12) The reaction was carried out according to the procedure outlined in Example 1. Method A, Step 9, tert-butyl 4-(6-((4-cyclopropyl-2-(2,4-dimethoxybenzyl)-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-7-yl)amino)pyridin-3-yl)piperazine-1-carboxylate (2.8) gave (1-12) as an orange solid. m / z=351 [M+H] + , 1 H NMR(400 MHz,DMSO):δ 9.54(s,1 H),9.01(s,2 H),7.97(d,J=2.5 Hz,1 H),7.43(dd,J=2.8,9.1 Hz,1 H),6.98(d,J=9.1 Hz,1 H),4.56(s,2 H),3.06-3.00(m,4 H),2.88(dd,J=4.5,4.5 Hz,4 H),2.25-2.18(br,1 H),2.09-2.01(m,1 H),1.04-0.93(m,4 H).

[0486] Example 3. Method C 4-(Methylsulfonyl)-7-((5-(piperazin-1-yl)pyridin-2-yl)amino)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one [ka] Synthesis of (I-5) Step 1: Ethyl 5-bromo-3-(dimethoxymethyl)-2-(methylthio)isonicotinate (3.0) To a solution of ethyl 5-bromo-2-chloro-3-(dimethoxymethyl)isonicotinate (1.5) (500 mg, 1.48 mmol) in DMF (15 mL) was added sodium thiomethoxide (114 mg, 1.62 mmol). After stirring at room temperature for 18 h, the mixture was quenched with NaHCO3 and extracted into DCM. The collected organic solution was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (0-50% gradient elution with EtOAc in isohexane) to give 3.0 (160 mg, 62%) as a colorless oil. 1 H NMR(400 MHz,CDCl3)δ 8.55(1H,d,J=2.0 Hz),5.44-5.42(1H,m),4.45-4.36(2H,m),3.45-3.36(6H,m),2.56(3H,s),1.42-1.36(3H,m).

[0487] Step 2. Ethyl 5-bromo-3-formyl-2-(methylthio)isonicotinate (3.1) The reaction was carried out according to the procedure ...

Claims

1. Compounds of Formula I 【Chemistry 81】 or a pharma- ceutically acceptable salt thereof, wherein: Z is CR or N; X is a covalent bond, -O-, -S-, -NR-, -S(O) ２ -, -S(O) ２ NR-, -S(O)-, -S(O)NR-, -C(O)-, -C(O)O-, -C(O)NR-, -C(O)N(R)O-, -OC(O)-, -OC(O)NR-, -N(R)C(O)O-, -N(R)C(O)-, -N(R)S(O) ２ - or X is C １－４ A divalent saturated or unsaturated, linear or branched hydrocarbon chain, wherein one or two methylene units of said chain are optionally and independently -C(R) ２ -, -N(R)-, -N(R)C(O)-, -C(O)N(R)-, -N(R)S(O) ２ -, -S(O) ２ N(R)-, -O-, -C(O)-, -OC(O)-, -C(O)O-, -S-, -S(O)-, or -S(O) ２ - has been replaced by R １ But, C １－６ aliphatic; phenyl; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocycle; a 5- to 6-membered monocyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and an 8- to 10-membered saturated or partially unsaturated bicyclic heterocycle having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from q R Ｃ has been replaced by the example R ２ But, C １－６ aliphatic; phenyl; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocycle; a 5- to 6-membered monocyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from q R Ｃ or R ２ -OR, -SR, -NR ２ , -S(O) ２ R, -S(O) ２ NR ２ 、-S(O)R、-S(O)NR ２ 、-C(O)R、-C(O)OR、-C(O)NR ２ 、-C(O)N(R)OR、 -OC(O)R、-OC(O)NR ２ 、-N(R)C(O)OR、-N(R)C(O)R、-N(R)C(O)NR ２ 、-N(R)C(NR)NR ２ 、 -N(R)NR ２ , -N(R)S(O) ２ N.R. ２ , -N(R)S(O) ２ R, -N=S(O)R ２ , —S(NR)(O)R, and —N(R)S(O)R; R ３ Each instance of is independently hydrogen or an optionally substituted C １－６ is an aliphatic group, R Ｃ Each instance of is independently oxo, halogen, -CN, -NO ２ , -OR, -SR, -NR ２ , -S(O) ２ R, -S(O) ２ NR ２ 、-S(O)R、-S(O)NR ２ 、-C(O)R、-C(O)OR、-C(O)NR ２ 、-C(O)N(R)OR、 -OC(O)R、-OC(O)NR ２ 、-N(R)C(O)OR、-N(R)C(O)R、-N(R)C(O)NR ２ 、-N(R)C(NR)NR ２ 、 -N(R)NR ２ , -N(R)S(O) ２ N.R. ２ , -N(R)S(O) ２ R, -N=S(O)R ２ , -S(NR)(O)R, -N(R)S(O)R, -N(R)CN, -P(O)(R)NR ２ , -P(O)(R)OR, or -P(O)R ２ or R Ｃ Each instance of １－６ aliphatic; phenyl; naphthalenyl; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocycle; a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, phosphorus, silicon, and sulfur; or a 5- to 6-membered monocyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; an 8- to 10-membered bicyclic heteroaryl ring having 1-5 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and s R are optionally substituted groups selected from: a 5-8 membered saturated or partially unsaturated bridged bicyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 6-11 membered saturated or partially unsaturated spiro ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or a 6-11 membered saturated or partially unsaturated bicyclic heterocycle having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from r instances of R and s instances of R. Ｄ has been replaced by the example R Ｄ Each instance of is independently oxo, halogen, -CN, -NO ２ , -OR, -SR-, NR ２ , -S(O) ２ R, -S(O) ２ NR ２ 、-S(O)R、-S(O)NR ２ 、-C(O)R、-C(O)OR、-C(O)NR ２ 、-C(O)N(R)OR、 -OC(O)R、-OC(O)NR ２ 、-N(R)C(O)OR、-N(R)C(O)R、-N(R)C(O)NR ２ 、-N(R)C(NR)NR ２ 、 -N(R)NR ２ , -N(R)S(O) ２ N.R. ２ , -N(R)S(O) ２ R, -N=S(O)R ２ , -S(NR)(O)R, -N(R)S(O)R, -N(R)CN, -P(O)(R)NR ２ , -P(O)(R)OR, or -P(O)R ２ and Each R is independently hydrogen, —CN, halogen, or C １－６ aliphatic; phenyl; naphthalenyl; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocycle; a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 5- to 6-membered monocyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; an 8- to 10-membered bicyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; an optionally substituted group selected from: a 7-12 membered saturated or partially unsaturated bicyclic heterocycle having heteroatoms; a 5-8 membered saturated or partially unsaturated bridged bicyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 6-10 membered saturated or partially unsaturated spirocycle having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 6-11 membered saturated or partially unsaturated bicyclic carbocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur; two R groups on the same nitrogen, taken together with said nitrogen, form an optionally substituted 4-7 membered monocyclic saturated, partially unsaturated, or heteroaryl ring having, in addition to said nitrogen, 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; an 8-10 membered bicyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 7-12 membered saturated or partially unsaturated bicyclic heterocycle having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; m is 0, 1, or 2; each q is independently 0, 1, 2, 3, or 4; each r is independently 0, 1, 2, 3, or 4; A compound of Formula I, wherein each s is independently 0, 1, 2, 3, or 4, or a pharma- ceutically acceptable salt thereof.

2. The compound is a compound of formula II 【Chemistry 82】 or a pharma- ceutically acceptable salt thereof.

3. The compound is a compound of formula V 【Chemistry 83】 or a pharma- ceutically acceptable salt thereof.

4. The compound is a compound of formula IV 【Chemistry 84】 or a pharma- ceutically acceptable salt thereof.

5. The compound is a compound of formula VII 【Chemistry 85】 or a pharma- ceutically acceptable salt thereof.

6. The compound of claim 1 , wherein X is —NR—.

7. R １ Let q be R Ｃ In the example, C is replaced １－６ Aliphatic; q R Ｃ phenyl substituted with q instances of R Ｃ q R Ｃ a 5-6 membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, substituted with, for example, q R Ｃ The compound of claim 1, wherein the ring is a 3-7 membered saturated or partially unsaturated monocyclic heterocycle having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, substituted with, for example,

8. R １ is phenyl or a 5-6 membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from q R Ｃ 8. The compound of claim 7, substituted with:

9. R １ phenyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, isothiazolyl, isoxazolyl, morpholinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl; -1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, oxetanyl, pyrimidinyl, piperazinyl, piperidinyl, pyranyl, pyrazinyl, piperyl, lazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, tetrahydrofuranyl, tetrahydropyranyl, thiazolyl, thienyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, azetidinyl, or xanthenyl, each of which is selected from the group consisting of q R Ｃ 2. The compound of claim 1 , substituted by, for example,

10. R １ is phenyl, pyrazolyl, pyridinyl, pyrazinyl, or pyrimidinyl, each of which is represented by q R Ｃ 10. The compound of claim 9 , substituted by, for example:

11. R １ but, 【Chemistry 86】 2. The compound of claim 1 ,

12. R １ but, 【Chemistry 87-1】 【Chemistry 87-2】 2. The compound of claim 1 ,

13. R ２ But, C １－６ aliphatic; phenyl; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocycle; a 5- to 6-membered monocyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from q R Ｃ or R ２ -OR, -SR, -NR ２ , -S(O) ２ R, -S(O) ２ N.R. ２ , -S(O)R, -S(O)NR ２ , -C(O)R, -C(O)OR, and -C(O)NR ２ 2. The compound of claim 1 , selected from:

14. R ２ is methyl, ethyl, n-propyl, i-Pr, n-Bu, s-Bu, t-Bu, straight-chain or branched pentyl, straight-chain or branched hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, phenyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, isothiazolyl, isoxazolyl, morpholinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl; 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, oxetanyl, pyrimidinyl, piperazinyl, piperidinyl nyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridinyl, pyridyl, pyridin-one, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, tetrahydropyranyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thiazolyl, thienyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, or azetidinyl, each of which is selected from the group consisting of q R Ｃ or R ２ -OR, -SR, -NR ２ , -S(O) ２ R, -S(O) ２ N.R. ２ , -S(O)R, -S(O)NR ２ , -C(O)R, -C(O)OR, and -C(O)NR ２ 14. The compound of claim 13, selected from:

15. R ２ is methyl, cyclopropyl, phenyl, imidazolyl, morpholinyl, oxazolyl, pyrazolyl, pyridazinyl, pyridinyl, pyridyl, pyridin-one, pyrimidinyl, pyrrolyl, or tetrahydropyranyl, each of which is selected from the group consisting of q R Ｃ or R ２ But -S(O) ２ R and -C(O)NR ２ 14. The compound of claim 13, selected from:

16. R ２ but, 【Chemistry 88】 14. The compound of claim 13,

17. R ２ But that R Ｃ Together with the substituents, 【Chemistry 89-1】 【Chemistry 89-2】 【Chemistry 89-3】 14. The compound of claim 13,

18. 2. The compound of claim 1, wherein the compound is selected from those shown in Table 1, or a pharma- ceutically acceptable salt thereof.

19. 13. A pharmaceutical composition comprising a compound of claim 1, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier, adjuvant, or vehicle.

20. 20. A composition comprising a compound according to claim 1 or a pharmaceutical composition according to claim 19 for use as a medicament.

21. 20. A method for inhibiting HPK1 in a biological sample, comprising contacting the sample with a compound of claim 1, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition of claim 19.

22. 20. A composition comprising a compound of claim 1, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition of claim 19, for treating a HPK1-mediated disorder, disease, or condition in a patient.

23. The composition of claim 22, wherein the disorder is a proliferative disorder.

24. 24. The composition of claim 23, wherein the proliferative disorder is cancer.

25. 24. The composition of claim 23, wherein the proliferative disorder is associated with one or more activating mutations in HPK1.

26. 23. The composition of claim 22, wherein the disease is a chronic viral infection.

27. A composition comprising a compound of claim 1, or a pharma- ceutically acceptable salt thereof, for increasing the efficacy of vaccination in a patient, or a pharmaceutical composition of claim 19, wherein the composition is administered as an adjuvant.

28. 20. Use of a compound of claim 1, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition of claim 19, in the manufacture of a medicament for treating an HPK1-mediated disorder, disease, or condition in a patient.

29. 29. The use according to claim 28, wherein the disorder is a proliferative disorder.

30. 30. The use of claim 29, wherein the proliferative disorder is cancer.

31. 30. The use of claim 29, wherein the proliferative disorder is associated with one or more activating mutations in HPK1.

32. 29. The use according to claim 28, wherein the disease is a chronic viral infection.

33. 20. Use of a compound according to claim 1, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition according to claim 19, in the manufacture of a medicament for increasing the effectiveness of vaccination in a patient as an adjuvant.

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