HPK1 antagonists and uses thereof

Abstract

The present invention provides compounds, compositions thereof, and methods of use thereof for the inhibition of HPK1 and the treatment of HPK1-mediated disorders.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 203,890, filed August 3, 2021, and U.S. Provisional Patent Application No. 63 / 264,749, filed December 1, 2021, each of which is incorporated by reference herein in its entirety.

[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 also 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 negatively regulates 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 is thought to result in 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. It is important to regulate the function of HPKl and immune cells, and is closely related to autoimmune diseases and anti-tumor immunity (Shui, JW, et al., Nat. Immunol, 2007.8(1):p.84-91;Wang,X.,et al.,J Biol Chem, 2012.287(14):p.11037-48)).

Prior Technical Literature

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[0005]

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

[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 for 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 described by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this invention, chemical elements are defined as defined in the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Edition (Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Edition). th Further, the general principles of organic chemistry are identified in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5 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, refer to 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 hybrids thereof, such as (cycloalkyl)alkenyl.

[0014] As used herein, the term "bridged bicyclic" refers to any saturated or partially unsaturated bicyclic ring system, i.e., carbocyclic or heterocyclic, 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 position" is any skeletal atom of the ring system that is bonded to three or more skeletal atoms (excluding hydrogen). In some embodiments, a 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 indicated for aliphatic groups. Additionally or alternatively, any substitutable nitrogen of a bridged bicyclic group may be substituted. Exemplary bridged bicyclic groups include: [ka]

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

[0016] The term "lower haloalkyl" refers to a straight or branched C alkyl group substituted with one or more halogen atoms. 1~4 Refers to an 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, "a saturated or unsaturated, straight or branched, divalent C 1~8 (or C 1~6 The term "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. The 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 hydrogen atoms have been 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 five to fourteen ring members, in which at least one ring in the system is aromatic and each ring in the system contains three to seven 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, for example, indanyl, phthalimidyl, naphthymidyl, phenanthridinyl, or tetrahydronaphthyl.

[0024] The terms "heteroaryl" and "heteroar-," 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 "heteroar-" 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 heteroaromatic ring or on one of the rings to which the heteroaromatic ring is fused. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, and tetrahydroisoquinolinyl. Heteroaryl groups can be monocyclic or bicyclic. The term "heteroaryl" may be used interchangeably with "heteroaryl ring," "heteroaryl group," or "heteroaromatic compound," any of which terms include optionally substituted rings. The term "heteroaralkyl" refers to an alkyl group substituted by a heteroaryl, where the alkyl and heteroaryl portions independently are optionally substituted.

[0025] As used herein, the terms "heterocycle," "heterocyclyl," "heterocyclic radical," and "heterocyclic ring" 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 has one or more, preferably one to four, heteroatoms in addition to carbon atoms, 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 0-3 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 can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure, and any of the ring atoms can 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 also 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 can 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 multiple positions in any given structure may be substituted with multiple substituents 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 permit 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 〇 ;-(CH2) 0~4 OR 〇 ;-O(CH2) 0~4 R o , -O-(CH2) 0~4 C(O)OR o ;-(CH2) 0~4 CH(OR 〇 )2;-(CH2) 0~4 SR 〇 ;R o may be substituted with -(CH2) 0~4 Ph;R o may be substituted with -(CH2) 0~4 O(CH2)0~1 Ph;R o may be substituted with -CH=CHPh; R o may be substituted with -(CH2) 0~4 O(CH2) 0~1 -pyridyl; -NO2; -CN; -N3; ​​-(CH2) 0~4 N(R 〇 )2;-(CH2) 0~4 N(R 〇 )C(O)R 〇 ;-N(R 〇 )C(S)R 〇 ;-(CH2) 0~4 N(R 〇 )C(O)NR 〇 2;-N(R 〇 )C(S)NR 〇 2;-(CH2) 0~4 N(R 〇 )C(O)OR 〇 ;-N(R 〇 )N(R 〇 )C(O)R 〇 ;-N(R 〇 )N(R 〇 )C(O)NR 〇 2;-N(R 〇 )N(R 〇 )C(O)OR 〇 ;-N(R 〇 )C(NR 〇 )N(R 〇 )2;-(CH2) 0~4 C(O)R 〇 ;-C(S)R 〇 ;-(CH2) 0~4 C(O)OR 〇 ;-(CH2) 0~4 C(O)SR 〇 ;-(CH2) 0~4 C(O)OSiR 〇 3;-(CH2) 0~4 OC(O)R 〇 ;-OC(O)(CH2) 0~4 SR o ;-SC(S)SR o ;-(CH2) 0~4 SC(O)R 〇 ;-(CH2) 0~4 C(O)NR 〇 2;-C(S)NR 〇2;-C(S)SR o ;-(CH2) 0~4 OC(O)NR 〇 2;-C(O)N(OR 〇 )R 〇 ;-C(O)C(O)R 〇 ;-C(O)CH2C(O)R 〇 ;-C(NOR 〇 )R 〇 ;-(CH2) 0~4 SSR 〇 ;-(CH2) 0~4 S(O)2R 〇 ;-(CH2) 0~4 S(O)2OR 〇 ;-(CH2) 0~4 OS(O)2R 〇 ;-S(O)2NR 〇 2;-(CH2) 0~4 S(O)R 〇 ;-N(R 〇 )S(O)NR 〇 2;-N(R 〇 )S(O)2R 〇 ;-N(OR 〇 )R 〇 ;-C(NH)NR 〇 2;-P(O)2R 〇 ;-P(O)R 〇 2;-OP(O)R 〇 2;-OP(O)(OR 〇 )2;-SiR 〇 3;-(straight or branched C 1~4 alkylene)ON(R 〇 )2; or -(linear or branched C 1~4 alkylene)C(O)ON(R 〇 )2, where each R 〇 may be substituted as defined below and independently represent hydrogen, C 1~6 Aliphatic, -CH2Ph, -O(CH2) 0~1 Ph, -CH2- (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, regardless of the above definition, two R 〇are taken together with their intervening atoms to 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] R 〇 The above suitable monovalent substituents (or two independently occurring R 〇 taken together with the intervening atoms) are independently halogen, -(CH2) 0~2 R 〇 ,-(Halo R ● ), -(CH2) 0~2 OH, -(CH2) 0~2 OR ● , -(CH2) 0~2 CH(OR ● )2;-O(HaloR ● ), -CN, -N3, -(CH2) 0~2 C(O)R ● , -(CH2) 0~2 C(O)OH, -(CH2) 0~2 C(O)OR ● , -(CH2) 0~2 SR ● , -(CH2) 0~2 SH, -(CH2) 0~2 NH2, -(CH2) 0~2 NHR ● , -(CH2) 0~2 NR ● 2, -NO2, -SiR ● 3. -OSiR ● 3. -C(O)SR ● , -(straight or branched C 1~4 alkylene)C(O)OR ● , or -SSR ● where each R ● is unsubstituted or, if preceded by "halo", substituted only with one or more halogens, and independently, C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0~1Ph, 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 include ═O 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 S-, wherein each R * is hydrogen, C which may be substituted as defined below 1~6 A suitable divalent substituent attached to adjacent substitutable carbon atoms of an "optionally substituted" group is selected from an aliphatic or unsubstituted 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. * 2) 2~3 O—, wherein each R * is hydrogen, C which may be substituted as defined below 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 ● ,-(Halo R ● ), -OH, -OR ● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2 or -NO2, wherein each R● is unsubstituted or, if preceded by "halo", substituted only with one or more halogens, and independently 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 † wherein each R † are independently hydrogen, 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, regardless of the above definition, two independently occurring R † together with their intervening atoms form an unsubstituted 3-12 membered saturated, partially unsaturated or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0034] R † Suitable substituents on the aliphatic groups are independently halogen, -R ● ,-(Halo R ● ), -OH, -OR ● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ●, -NH2, -NHR ● , -NR ● 2, or -NO2, where each R ● is unsubstituted or, if preceded by "halo", is substituted with only one or more halogens, and independently, 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, 2-hydroxy-ethanoate, and the like. Examples of the salts include benzoate, benzoyl persulf ...

[0036] Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, and N+ (C 1~4 Representative 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, for example, 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 isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational) forms of the structure; for example, R and S configurations about each chiral center, Z and E double bond isomers, and Z and E conformational isomers. Therefore, 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 tautomeric forms 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 in accordance with 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] As used herein, an "HPK1 antagonist" or "HPK1 inhibitor" is a molecule that reduces, inhibits, or otherwise decreases one or more of the biological activities of HPK1 (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 imply 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 the phosphorylation of SLP76 and / or Gads by HPK1. Compounds of the present disclosure directly bind to HPK1 and inhibit its kinase activity.

[0039] 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.

[0040] The compounds of the present invention can be linked to a detectable moiety. It will be understood that such compounds are useful as imaging reagents. Those skilled in the art will recognize that the detectable moiety can be attached to the provided compounds via a suitable substituent. As used herein, the term "suitable substituent" refers to a moiety that can be covalently attached to a detectable moiety. Such moieties are well known to those skilled in the art and include, for example, groups containing a carboxylic acid moiety, an amino moiety, a thiol moiety, or a hydroxyl moiety, to name a few. It will be understood that the moiety can be attached to the provided compounds directly or via a linking group such as a saturated or unsaturated divalent hydrocarbon chain. In some embodiments, such moieties can be attached via click chemistry. In some embodiments, such moieties can 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 are described in Rostovtsev et al., Angew. Chem. Int. Ed. 2002, 41,2596-99, and Sun et al.,Bioconjugate Chem.,2006, 17 ,52-57.

[0041] 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 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.

[0042] 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. In the case of biotin, the secondary intermediate may include a streptavidin-enzyme conjugate. In the case of 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.

[0043] 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'-demethoxy-fluorescein, DM-NERF, eosin, erythrosine, fluorescein, FAM, hydroxycoumarin, IRDye dyes (IRD40, IRD700, IRD800), JOE, Lissamine rhodamine B, Marina 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.

[0044] 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.

[0045] 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.

[0046] 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 a saturated or unsaturated straight or branched divalent C 1~4 a hydrocarbon chain, wherein one or two methylene units of the chain are optionally and independently replaced by -C(R)-, -N(R)-, -N(R)C(O)-, -C(O)N(R)-, -N(R)S(O)-, -S(O)N(R)-, -O-, -C(O)-, -OC(O)-, -C(O)O-, -S-, -S(O)-, or -S(O)-; R 1 is 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 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and an 8- to 11-membered saturated or partially unsaturated bicyclic heterocycle having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from R for an instance of q. C is replaced by R 2 is a 6- to 11-membered saturated, partially unsaturated, or unsaturated fused, bridged, or spiro bicyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from the R of an instance of q. c is replaced by R 3 Each instance of is independently hydrogen or an optionally substituted C 1~6 is an aliphatic group, R CEach 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; independently nitrogen, an optionally substituted group 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 spirocyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or a 6-11 membered saturated or partially unsaturated bicyclic heterocyclic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is independently selected from R at an instance of r and R at an instance of s. D is replaced by R DEach instance of is independently selected from 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 1~6 Aliphatic; 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 to 3 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 carbocycle having 1 to 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; and Compounds of Formula I, or pharmaceutically acceptable salts thereof, are provided wherein each s is independently 0, 1, 2, 3, or 4.

[0047] Z is CR or N, as generally defined above.

[0048] In some embodiments, Z is CR. In some embodiments, Z is N.

[0049] In some embodiments, Z is CH.

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

[0051] 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)—.

[0052] In some embodiments, X is a saturated or unsaturated, linear or branched, divalent C 1~4A hydrocarbon chain wherein one or two methylene units of the chain are optionally and independently replaced with -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-.

[0053] 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-.

[0054] In certain embodiments, X is -NR-. In certain embodiments, X is -NH-.

[0055] In some embodiments, X is selected from those shown in Table 1 below.

[0056] In some embodiments, R 1 is the R of the instance of q C C replaced by 1~6 Aliphatic; q instance R C q instances of R C q is an instance of a saturated or partially unsaturated monocyclic carbocyclic ring substituted with R C a 5-6 membered monocyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, substituted with 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 C and R is an 8- to 11-membered saturated or partially unsaturated bicyclic heterocycle having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, substituted with

[0057] In some embodiments, R1 is phenyl or a 5-6 membered monocyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from R at an instance of q. C is replaced by

[0058] In certain embodiments, R 1is phenyl, indanyl, tetrahydronaphthyl, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benzotriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, NH-carbazolyl, carbolinyl, chromanyl, chromophenyl ... Chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isoindolinyl, isoindolenyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinoline Linyl, 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, phenoxathiinyl, 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, 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 an instance of R, C is replaced by

[0059] In certain embodiments, R 1 is 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, pyrazo lysinyl, 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 an instance of R C is replaced by

[0060] In certain embodiments, R 1is 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 an instance of R C is replaced by

[0061] In certain embodiments, R 1 is phenyl, pyrazolyl, pyridinyl, pyrazinyl, or pyrimidinyl, each of which is an instance of q of R C is replaced by

[0062] In certain embodiments, R 1 teeth, [ka] is.

[0063] In certain embodiments, R 1 teeth, [ka] and In the formula, R C Each instance of is independently -OR, -NR2, C 1~6 an aliphatic, 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, each of which is an R of an instance of r and an R of an instance of s; D is replaced by

[0064] In certain embodiments, the R C R together with the substituent 1 teeth, [ka] [ka] [ka] [ka] is.

[0065] In some embodiments, R 1 is selected from those shown in Table 1 below.

[0066] Generally as defined above, R 2 is a 6- to 11-membered saturated, partially unsaturated, or unsaturated fused, bridged, or spiro bicyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is selected from the R of an instance of q. c is replaced by

[0067] In some embodiments, R 2 is a 6- to 11-membered saturated, partially unsaturated, or unsaturated fused, bridged, or spiro bicyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and q is an instance of R C is replaced by

[0068] In certain embodiments, R 2 is a 7-10 membered fused bicyclic ring having 1-3 nitrogen atoms, each of which is an R at an instance of q. C is replaced by each of those replaced by

[0069] In certain embodiments, R 2 is a 9-membered fused bicyclic ring having 1 to 3 nitrogen atoms, each of which is an R at an instance of qC wherein each R C are independently halogen, —CN, —OR, —C(O)NR, —NR, or R C Each instance of C 1~6 an optionally substituted group selected from aliphatic; phenyl; 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; and a 6- to 11-membered saturated or partially unsaturated fused, bridged, or spiro bicyclic heterocycle having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; C Each instance of is independently a R of instances of r and a R of instances of s. D is optionally replaced by

[0070] In certain embodiments, R 2 teeth, [ka] is.

[0071] In certain embodiments, the R C R together with the substituent 2 teeth, [ka] is.

[0072] In some embodiments, R 2 is selected from those shown in Table 1 below.

[0073] Generally as defined above, R 3 Each instance of is independently hydrogen or an optionally substituted C 1~6 It is an aliphatic group.

[0074] In some embodiments, R 3 is hydrogen.

[0075] In some embodiments, R 3 is selected from those shown in Table 1 below.

[0076] Generally as defined above, R C Each instance of is independently a halogen, -CN, or -C(O)R, or R C Each instance of C 1~6 an optionally substituted group selected from aliphatic; a 3- to 7-membered saturated or partially unsaturated monocyclic carbocyclic ring; or 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, each of which is R at an instance of r and R at an instance of s. D is replaced by

[0077] In some embodiments, R C is methyl, ethyl, isopropyl, cyclopentyl, —CN, fluoro, or methoxy.

[0078] In some embodiments, R C teeth, [ka] is.

[0079] In some embodiments, R C is -CHF2 or chloro or fluoro.

[0080] In some embodiments, R C teeth [ka] is.

[0081] In some embodiments, R C teeth, [ka] is.

[0082] In some embodiments, R C Each instance of is selected from those shown in Table 1 below.

[0083] As outlined above, R D Each instance of is independently oxo, -OR, or -NR2.

[0084] In some embodiments, R D is hydroxy, fluoro, or methoxy.

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

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

[0087] In some embodiments, each R is independently hydrogen or C 1~6 An optionally substituted group selected from aliphatic; 3- to 7-membered saturated or partially unsaturated monocyclic carbocyclic ring; and 3- to 7-membered saturated or partially unsaturated monocyclic heterocyclic ring having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0088] In some embodiments, R is hydrogen, methyl, [ka] is.

[0089] In some embodiments, R is selected from those shown in Table 1 below.

[0090] As generally defined above, each hydrogen bonded to a carbon can be optionally and independently replaced with deuterium.

[0091] In some embodiments, the hydrogen bonded to the carbon is replaced with deuterium.

[0092] As generally defined above, m is 0, 1, or 2.

[0093] In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2.

[0094] In some embodiments, m is selected from those shown in Table 1 below.

[0095] 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.

[0096] In some embodiments, q is 1, 2, or 3. In some embodiments, q is 1 or 2.

[0097] In some embodiments, q is selected from those shown in Table 1 below.

[0098] 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.

[0099] 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.

[0100] In some embodiments, r is selected from those shown in Table 1 below.

[0101] 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.

[0102] 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.

[0103] In some embodiments, s is selected from those shown in Table 1 below.

[0104] 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 which, both alone and in combination, is as defined above and described in the embodiments herein.

[0105] 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 which, both alone and in combination, is as defined above and described in the embodiments herein.

[0106] 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 which, both alone and in combination, is as defined above and described in the embodiments herein.

[0107] 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.

[0108] 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 which, both alone and in combination, is as defined above and described in the embodiments herein.

[0109] 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 which, both alone and in combination, is as defined above and described in the embodiments herein.

[0110] In some embodiments, the present invention provides a compound of formula VIII-a or VIII-b: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R C , X, and q, both alone and in combination, are as defined above and in the embodiments herein.

[0111] In some embodiments, the present invention provides a compound of formula IX-a or IX-b: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R c and q, both alone and in combination, are as defined above and in the embodiments herein.

[0112] In some embodiments, the present invention provides a compound of formula Xa or Xb: [ka] or a pharmaceutically acceptable salt thereof, wherein R C and q, both alone and in combination, are as defined above and in the embodiments herein.

[0113] In some embodiments, the present invention provides a compound of formula XI-a or XI-b: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R c , X, and q, both alone and in combination, are as defined above and in the embodiments herein.

[0114] In some embodiments, the present invention provides a compound of formula XII-a or XII-b: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R c and q, both alone and in combination, are as defined above and in the embodiments herein.

[0115] In some embodiments, the present invention provides a compound of formula XIII-a or XIII-b: [ka] or a pharmaceutically acceptable salt thereof, wherein R C and q, both alone and in combination, are as defined above and in the embodiments herein.

[0116] In some embodiments, the present invention provides a compound of formula XIV-a or XIV-b: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R c , X, and q, both alone and in combination, are as defined above and in the embodiments herein.

[0117] In some embodiments, the present invention provides a compound of formula XV-a or XV-b: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R c , X, and q, both alone and in combination, are as defined above and in the embodiments herein.

[0118] In some embodiments, the present invention provides a compound of formula XVI-a or XVI-b: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R c , X, and q, both alone and in combination, are as defined above and in the embodiments herein.

[0119] 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]

[0120] 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.

[0121] 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.

[0122] 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.

[0123] 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.

[0124] 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.

[0125] 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.

[0126] 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, for enhancing an immune response in a subject in need thereof, and / or for treating an HPK1-dependent disorder.

[0127] 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.

[0128] 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.

[0129] 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.

[0130] 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.

[0131] 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.

[0132] 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.

[0133] 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.

[0134] 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.

[0135] 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.

[0136] The term "patient", as used herein, means an animal, preferably a mammal, and most preferably a human.

[0137] 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 to be 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.

[0138] "Pharmaceutically acceptable derivative" means any non-toxic salt, ester, salt of an ester, or other derivative of a compound of this invention which 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.

[0139] As used herein, the term "inhibitorily active metabolite or residue thereof" means that the metabolite or residue thereof is also an inhibitor of HPK1 or a mutant thereof.

[0140] 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 base 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.

[0141] The compounds of the present invention may be in the form of a "prodrug," which includes compounds having a moiety that can be metabolized in vivo. Typically, 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 the purified compound in its free acid form or in a hydroxyl group with a suitable esterifying agent. The 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.

[0142] 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 "parenteral" 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. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium.

[0143] 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 forms, 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 Tween®, Span®, 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.

[0144] 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 emulsifying and suspending agents.If desired, certain sweeteners, flavorings or coloring agents can also be added.

[0145] Alternatively, the pharmaceutically acceptable compositions of the present invention can 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, thereby melting in the rectum to release the drug. Such materials include cocoa butter, beeswax, and polyethylene glycol.

[0146] 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.

[0147] 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.

[0148] For topical application, the provided pharmaceutically acceptable compositions may 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 paraffin, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsifying wax, and water. Alternatively, the provided pharmaceutically acceptable compositions may be formulated into a suitable lotion or emulsion 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 esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and water.

[0149] For ophthalmic use, the provided pharmaceutically acceptable compositions may be formulated as a micronized suspension in isotonic, pH-adjusted, sterile saline, or preferably as a solution in isotonic, pH-adjusted, sterile saline, with or without a preservative, such as benzylalkonium chloride. Alternatively, for ophthalmic use, the pharmaceutically acceptable compositions may be formulated in an ointment, such as petrolatum.

[0150] 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.

[0151] Most preferably, pharmaceutically acceptable compositions of the present invention are formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, pharmaceutically acceptable compositions of the present invention are administered without food. In other embodiments, pharmaceutically acceptable compositions of the present invention are administered with food.

[0152] 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 upon the host treated, the particular mode of administration, etc. 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.

[0153] 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 patient's age, body weight, general health, sex, diet, time of administration, excretion rate, concomitant medications, 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 specific compound in the composition.

[0154] 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.

[0155] 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.

[0156] 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.

[0157] 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 exhibit at least one of enhanced priming, enhanced activation, enhanced migration, enhanced proliferation, enhanced survival, and enhanced cytolytic activity compared to before administration of the compound or pharmaceutical composition. In certain aspects of this embodiment, T cell activation is characterized by an increased frequency of γ-IFN+ CD8 T cells or enhanced levels of IL-2 or granzyme B production by the T cells compared to before administration of the compound or pharmaceutical composition. In certain aspects of this embodiment, the number of T cells is increased compared 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 exhibit enhanced maturation and activation compared 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 increase in the frequency of CD83+ dendritic cells. In certain aspects of this embodiment, antigen-presenting cell activation is characterized by an increase in the expression of CD80 and CD86 on dendritic cells.

[0158] Compounds of the present disclosure directly bind to HPK1 and inhibit its kinase activity, hi some embodiments, compounds of the present disclosure reduce, inhibit, or otherwise decrease the phosphorylation of SLP76 and / or Gads by HPK1.

[0159] 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.

[0160] 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 HPK1 and inhibit its activity. The compound can be contacted with HPK1 in vitro or in vivo via administration of the compound to a subject.

[0161] 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.

[0162] 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.

[0163] 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.

[0164] 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 proliferation of T cells (e.g., CD4 T cells, CD8 T cells), 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 production of cytokines (e.g., interleukin-2) by T cells, increased resistance to prostaglandin E2-induced immunosuppression, and / or enhanced priming and / or cytolytic activity of CD8 T cells.

[0165] In some embodiments, CD8 T cells in a subject exhibit enhanced priming, activation, proliferation, and / or cytolytic activity compared to before 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 increased CD44 expression and / or enhanced cytolytic activity on CD8 T cells. In some embodiments, CD8 T cell activation is characterized by increased γ-IFN expression and / or increased cytolytic activity on CD8 T cells. + It is characterized by an increased frequency of CD8 T cells. In some embodiments, the CD8 T cells are antigen-specific T cells.

[0166] In some embodiments, antigen-presenting cells in a subject are enhanced in 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, maturation of antigen-presenting cells is enhanced by CD83 + In some embodiments, activation of antigen-presenting cells is characterized by increased expression of CD80 and CD86 on dendritic cells.

[0167] In some embodiments, the serum levels of the cytokine IL-10 and / or the chemokine IL-8, 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.

[0168] TCR engagement leads to the 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, which leads to T cell dysfunction, including anergy and exhaustion (Lasserre et al. (2011) J Cell Biol 195(5):839-853).

[0169] 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.

[0170] 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 decreased 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 decreased ability to secrete cytokines, proliferate, or achieve cytolytic activity. In certain aspects, the decreased 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.

[0171] Thus, the compounds of the present disclosure can be used in the treatment of conditions where enhanced immunogenicity is desired, such as increasing tumor immunogenicity for the treatment of cancer.

[0172] The term "dysfunction" in the context of immune dysfunction refers to a state of reduced immune responsiveness to antigenic stimulation. This term includes the common elements of both exhaustion and / or anergy, such that antigen recognition may occur but the subsequent immune response is ineffective in controlling infection or tumor growth.

[0173] 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.

[0174] 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.

[0175] 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 distinct from anergy in that it results from persistent signaling, rather than from incomplete or defective signaling. It is defined by insufficient effector function, persistent expression of inhibitory receptors, and a transcriptional state that differs from that of 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 cell-intrinsic negative regulatory (costimulatory) pathways (PD-1, B7-H3, B7-H4, etc.).

[0176] "Immunogenicity" refers to the ability of a particular substance to induce an immune response. Because tumors are immunogenic, enhancing tumor immunogenicity helps eliminate tumor cells through the immune response.

[0177] "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 (e.g., clearance of viruses, pathogens, or tumors) compared to such levels before the intervention, 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.

[0178] "Tumor immunity" refers to the process by which tumors evade immune recognition and elimination. 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 elimination.

[0179] 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.

[0180] 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.

[0181] In the methods described herein, a compound of the present invention or a pharmaceutical composition thereof is administered to a subject with cancer.

[0182] 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, cancer cells in the subject selectively increase expression of MHC class I antigen expression compared to before administration of the compound or composition.

[0183] 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.

[0184] 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.

[0185] 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.

[0186] In certain embodiments, the cell proliferative disorder is cancer.

[0187] 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 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 endometrium, endometrial cancer, cervical cancer, vaginal cancer, cancer of the vulva, Hodgkin's disease, 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, soft tissue sarcoma, cancer of the urethra, cancer of the penis, and acute myeloid leukemia. , chronic or acute leukemia including chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, childhood solid tumors, 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 cancer, and combinations of such cancers.

[0188] 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, renal cancer, liver cancer, pancreatic cancer, gastric cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, sarcoma, bladder cancer, etc.), hematological cancers (e.g., lymphoma, leukemia such as 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.

[0189] In certain embodiments, the cancer is brain cancer, leukemia, skin cancer, prostate cancer, thyroid cancer, colon cancer, lung cancer, or sarcoma. In other embodiments, the cancer is selected from the group consisting of glioma, glioblastoma multiforme, paraganglioma, suprantentorial primordial 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.

[0190] 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 tract cancer, and cancer of unknown primary origin (cancer that has metastasized but the site of the primary 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.

[0191] 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 estheosioneuroblastoma (olfactory neuroblastoma). In certain embodiments, the brain cancer is selected from the group consisting of astrocytoma (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), oligodendroglioma (e.g., oligodendroglioma and anaplastic oligodendroglioma), oligoastrocytic tumor ( For example, the brain cancer is selected from the group consisting of oligodendroglioma, and anaplastic oligodendroglioma), ependymoma (e.g., myxopapillary ependymoma, and anaplastic ependymoma); medulloblastoma, primitive neuroectodermal tumor, schwannoma, meningioma, atypical meningioma, anaplastic meningioma, pituitary adenoma, brain stem glioma, cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, visual pathway and hypothalamic glioma, and primary central nervous system lymphoma. In certain examples of these embodiments, the brain cancer is selected from the group consisting of glioma, glioblastoma multiforme, paraganglioma, and suprantentorial primitive neuroectodermal tumor (sPNET).

[0192] 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. In certain embodiments, the eye cancer is selected from the group consisting of intraocular melanoma and retinoblastoma.

[0193] 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 neoplasm, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), chronic myeloid leukemia (CML), myeloproliferative neoplasm (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 plasmacytoma, including plasmacytoma and multiple myeloma. The leukemias referred to herein can be acute or chronic.

[0194] 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.

[0195] 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.

[0196] 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.

[0197] 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.

[0198] In certain embodiments, the cancer is selected from renal cancer and bladder cancer. In certain embodiments, the cancer is renal 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 urethelial carcinoma (transitional cell carcinoma), squamous cell carcinoma, and adenocarcinoma.

[0199] 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.

[0200] 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.

[0201] In certain embodiments, the cancer is selected from the group consisting of malignant mesothelioma, epithelioid mesothelioma, and sarcoid.

[0202] 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.

[0203] In certain embodiments, the cancer is selected from lymphoma. 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.

[0204] 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.

[0205] 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.

[0206] 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.

[0207] 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, rhabdomyosacroma, angiosarcoma, chondrosarcoma, infantile fibrosarcoma, and synovial sarcoma.

[0208] 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 neoplasms (e.g., mole destructive, choriocarcinoma, placental site trophoblastic tumor, and epithelioid trophoblastic tumor).

[0209] In some embodiments, the subject has melanoma. The melanoma can be early or late stage. In some embodiments, the subject has colorectal cancer. The colorectal cancer can be early or late stage. In some embodiments, the subject has non-small cell lung cancer. The non-small cell lung cancer can be early or late stage. In some embodiments, the subject has pancreatic cancer. The pancreatic cancer can be in an early or late stage state. In some embodiments, the subject has a hematological malignancy. The hematological malignancy can be early or late stage. In some embodiments, the subject has ovarian cancer. The ovarian cancer can be early or late stage. In some embodiments, the subject has breast cancer. The breast cancer can be early or late stage. In some embodiments, the subject has renal cell carcinoma. The renal cell carcinoma can be early or late stage. In some embodiments, the cancer has an elevated level of T cell infiltration.

[0210] In some embodiments, cancers treatable with the compounds of the present disclosure include melanoma (e.g., metastatic malignant melanoma), renal 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 refractory or recurrent malignancies whose growth can be inhibited using the compounds of the present disclosure.

[0211] 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.

[0212] 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.

[0213] Exemplary sarcomas include chondrosarcoma, Ewing's sarcoma, osteosarcoma, rhabdomyosarcoma, angiosarcoma, fibrosarcoma, liposarcoma, myxoma, rhabdomyoma, rhabdomyosarcoma, fibroma, lipoma, hamartoma, and teratoma.

[0214] Exemplary lung cancers include non-small cell lung cancer (NSCLC), small cell lung cancer, bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, chondroitin hamartoma, and mesothelioma.

[0215] 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 cancer (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), colon cancer (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma), and colorectal cancer.

[0216] Exemplary genitourinary cancers include cancer of the kidney (adenocarcinoma, Wilms' tumor [nephroblastoma]), cancer of the bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate cancer (adenocarcinoma, sarcoma), and testicular cancer (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell tumor, fibroma, fibroadenoma, adenomatous tumor, lipoma).

[0217] Exemplary liver cancers include hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, and hemangioma.

[0218] 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 tumor chordoma, osteochronfroma (osteocartilaginous exostoses), benign chondroma, chondroblastoma, chondromyxoid fibroma, osteoid osteoma, and giant cell tumor.

[0219] 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, germ cell 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-Dacros disease.

[0220] Exemplary gynecological cancers include cancer of the uterus (endometrial carcinoma), cancer of the cervix (cervical carcinoma, pre-tumor cervical dysplasia), cancer of the ovaries (ovarian carcinoma (serous cystadenocarcinoma, 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).

[0221] 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.

[0222] 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.

[0223] 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 colorectal cancer, breast cancer, pancreatic cancer, and lung cancer, and 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, elevated CD73 expression levels are associated with tumor neovascularization, invasiveness, and metastasis, and shorter patient survival in breast cancer patients.

[0224] 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, 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.

[0225] 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 their pharmaceutically acceptable salts, prodrugs, metabolites, or derivatives are administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intracerebroventricularly, intratumorally, or intranasally.

[0226] 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.

[0227] It is understood that the appropriate dose of the active compound depends on several factors within the knowledge of a physician or veterinarian of ordinary skill in the art, and 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 concomitant medications.

[0228] It will also be understood that the effective dosage of a compound of the invention or a pharmaceutically acceptable salt, prodrug, metabolite, or derivative thereof used for treatment may increase or decrease over the course of a particular treatment. Modifications in dosage may occur and be apparent from the results of diagnostic assays.

[0229] In some embodiments, the HPK1 antagonist is administered to a subject at a dose of about 0.001 μg / kg 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.

[0230] 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.

[0231] As used herein, the terms "treatment," "treat," and "treating" refer to reversing, alleviating, delaying the onset of, or inhibiting 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.

[0232] 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.

[0233] 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.

[0234] 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 metabolism and other similar processes.

[0235] 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 of an agent 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. It may be cytostatic and / or cytotoxic, to the extent that the agent can prevent growth and / or kill existing cancer cells. In the case of cancer treatment, efficacy can be measured, for example, by assessing the time to disease progression (TTP) and / or determining the response rate (RR).

[0236] 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.

[0237] 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 to be "appropriate for the disease, or condition, being treated."

[0238] In certain embodiments, a provided combination, or composition thereof, is administered in combination with another therapeutic agent.

[0239] Examples of drugs that may be combined in combination with the agents of the present invention include, but are not limited to, drugs for treating Alzheimer's disease such as Aricept® and Excelon®; drugs for treating HIV such as ritonavir; drugs for treating Parkinson's disease such as L-DOPA / carbidopa, entacapone, ropinrole, pramipexole, bromocriptine, pergolide, trihexephendyl, and amantadine. medications for treating multiple sclerosis (MS), such as beta interferons (e.g., Avonex® and Rebif®), Copaxone®, and mitoxantrone; medications for treating asthma, such as albuterol and Singulair®; medications for treating schizophrenia, such as Zyprexa, Risperdal, Seroquel, and haloperidol; corticosteroids, TNF inhibitors, IL-1 Anti-inflammatory drugs such as RA, azathioprine, cyclophosphamide, and sulfasalazine; immunomodulators and immunosuppressants such as cyclosporine, tacrolimus, rapamycin, mycophenolate mofetil, interferons, corticosteroids, cyclophophamide, azathioprine, and sulfasalazine; neurotrophic factors such as acetylcholinesterase inhibitors, MAO inhibitors, interferons, anticonvulsants, ion channel blockers, riluzole, and antiparkinsonian drugs; beta-blockers, ACE inhibitors, diuretics, nitrates, These include drugs for treating cardiovascular disease, such as calcium channel blockers and statins; drugs for treating liver disease, such as corticosteroids, cholestyramine, interferons, and antivirals; drugs for treating blood disorders, such as corticosteroids, anti-leukemias, 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.

[0240] 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.

[0241] These additional agents may be administered separately from the provided combination therapy as part of a multiple dose regimen. Alternatively, the agents may be part of a single dosage form, mixed together with the compounds of this invention in a single composition. When administered as part of a multiple dose regimen, the two active agents may be given simultaneously, sequentially, or within a period of each other, usually within five hours of each other.

[0242] As used herein, the terms "co-administration," "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.

[0243] The amount of additional therapeutic agent present in the compositions of the present invention will be 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 will be in the range of about 50% to 100% of the amount that would normally be present in a composition comprising that agent as the only therapeutically active agent.

[0244] 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.

[0245] 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, acetaminophen, nonsteroidal anti-inflammatory drugs (NSAIDS) such as aspirin, ibuprofen, naproxen, etodolac (Lodine®), and celecoxib, colchicine (Colcrys®), corticosteroids such as prednisone, prednisolone, methylprednisolone, hydrocortisone, probenecid, allopurinol, febuxostat (Uloric®), and the like. Other drugs include benzodiazepines (Benza, Bifidobacterium ... Drugs that are not approved for use in the treatment of rheumatoid arthritis include steroids (TNF-α, steroids, anti-inflammatory drugs ... ), infliximab (Remicade®), golimumab (Simponi®), certolizumab pegol (Cimzia®), and adalimumab (Humira®); "anti-IL-1" agents such as anakinra (Kineret®) and rilonacept (Arcalyst®); canakinumab (Ilaris®); anti-Jak inhibitors such as 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®), antidiarrheal drugs such as diphenoxylate (Lomotil®) and loperamide (Imodium®); bile acid binders such as cholestyramine, alosetron (Lotronex®), lubiprostone (Amitiza®); laxatives such as milk of magnesia, polyethylene glycol (MiraLax®), Dulcolax®, Correctol®, and Senokot®; anticholinergics or antispasmodics such as dicyclomine (Bentyl®), Singulair®, beta-2 agonists such as albuterol (Ventolin® HFA, Proventil® HFA), levalbuterol (Xopenex®), metaproterenol (Alupent®), pirbuterol acetate (Maxair®), terbutaline sulfate (Brethaire®), serotonin-releasing hormone (Senokot®), etc. anticholinergics such as meterol xinafoate (Serevent®) and formoterol (Foradil®), ipratropium bromide (Atrovent®) and tiotropium (Spiriva®), inhaled corticosteroids such as beclomethasone dipropionate (Beclovent®, Qvar®, and Vanceril®), triamcinolone acetonide (Azmacort®), mometasone (Asthmanex®),Budesonide (Pulmocort®), and flunisolide (Aerobid®), such as Afviar®, Symbicort®, Dulera®, cromolyn sodium (Intal®), methylxanthines such as theophylline (Theo-Dur®, Theolair®, Slo-bid®, Uniphyl®, Theo-24®), and aminophylline IgE antibodies such as omalizumab (Xolair®), nucleoside reverse transcriptase inhibitors such as zidovudine (Retrovir®), abacavir (Ziagen®), abacavir / lamivudine (Epzicom®), abacavir / lamivudine / zidovudine (Trizivir®), didanosine (Videx®), emtricitabine (Emtriva®), lamivudine (Epivir®), lamivudine (Trizi ... non-nucleoside reverse transcriptase inhibitors such as delavirdine (Rescriptor®), efavirenz (Sustiva®), nevirapine (Viramune®), and etravirine (Intelence®), nucleotide reverse transcriptase inhibitors such as tenofovir (Viread®), protease inhibitors such as vudine / zidovudine (Combivir®), stavudine (Zerit®), and zalcitabine (Hivid®); antivirals such as amprenavir (Agenerase®), atazanavir (Reyataz®), darunavir (Prezista®), fosamprenavir (Lexiva®), indinavir (Crixivan®), lopinavir and ritonavir (Kaletra®), nelfinavir (Viracept®), ritonavir (Norvir®), saquinavir (Fortovase® or Invirase®),and entry inhibitors such as tipranavir (Aptivus®), enfuvirtide (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.

[0246] In another embodiment, the present invention provides a method of treating rheumatoid arthritis by administering to a patient in need thereof a compound of Formula I and one or more of the following: nonsteroidal anti-inflammatory drugs (NSAIDS) such as aspirin, ibuprofen, naproxen, etodolac (Lodine®), and celecoxib; corticosteroids such as prednisone, prednisolone, methylprednisolone, hydrocortisone, sulfasalazine (Azulfidine®); antimalarials such as hydroxychloroquine (Plazolinone); These include methotrexate (Rheumatrex®), gold salts such as gold thioglucose (Solganal®), gold thiomalate (Myochrysine®), and auranofin (Ridaura®), D-penicillamine (Depen® or Cuprimine®), azathioprine (Imuran®), cyclosporin (Campan ... Rofosfamide (Cytoxan®), chlorambucil (Leukeran®), cyclosporine (Sandimmune®), leflunomide (Arava®), and "anti-TNF" agents such as etanercept (Enbrel®), infliximab (Remicade®), golimumab (Simponi®), certolizumab pegol (Cimzia®), and amphetamine (Amphetamine®). and administering one or more additional therapeutic agents selected from "anti-IL-1" agents such as dalimumab (Humira®), including anakinra (Kineret®) and rilonacept (Arcalyst®), antibodies such as rituximab (Rituxan®), "anti-T cell" agents such as abatacept (Orencia®), and "anti-IL-6" agents such as tocilizumab (Actemra®).

[0247] In some embodiments, the present invention provides methods 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, nonsteroidal anti-inflammatory drugs (NSAIDS) such as aspirin, ibuprofen, naproxen, etodolac (Lodine®), and celecoxib, diclofenac, cortisone, hyaluronic acid (Synvisc® and Hyalgan®), and monoclonal antibodies such as tanezumab.

[0248] In some embodiments, the present invention provides a method of treating cutaneous lupus erythematosus or systemic lupus erythematosus by administering to a patient in need thereof a compound of Formula I and one or more of the following combinations: acetaminophen; nonsteroidal anti-inflammatory drugs (NSAIDS) such as aspirin, ibuprofen, naproxen, etodolac (Lodine®), and celecoxib; corticosteroids such as prednisone, prednisolone, methylprednisolone, hydrocortisone; or antimalarials such as hydroxychloroquine (Plaquenil). (Rheumatrex®), azathioprine (Imuran®), and anticoagulants such as heparin (Calcinparine® or Liquaemin®) and warfarin (Coumadin®).

[0249] In some embodiments, the present invention provides a method of treating Crohn's disease, ulcerative colitis, or inflammatory bowel disease by administering to a patient in need thereof a compound of Formula I and one or more of the following: mesalamine (Asacol®), sulfasalazine (Azulfidine®), antidiarrheals such as diphenoxylate (Lomotil®) and loperamide (Imodium®), bile acid binding drugs such as cholestyramine, alosetron (Lotronex®), rubin (Rubiconazole), cyclosporine (Cyclosporin ... and administering one or more additional therapeutic agents selected from prostones (Amitiza®), laxatives such as 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.

[0250] 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 an anticholinergic agent such as Singulair (Singulair®), beta-2 agonists such as albuterol (Ventolin® HFA, Proventil® HFA), levalbuterol (Xopenex®), metaproterenol (Alupent®), pirbuterol acetate (Maxair®), terbutaline sulfate (Brethaire®), salmeterol xinafoate (Serevent®), and formoterol (Foradil®), ipratropium bromide (Atrovent®) and tiotropium (Spiriva®), an inhaled corticosteroid such as prednisone, prednisolone, beclomethasone dipropionate (Bec and administering one or more additional therapeutic agents selected from: lovent®, Qvar®, and Vanceril®), triamcinolone acetonide (Azmacort®), mometasone (Asthmanex®), budesonide (Pulmocort®), flunisolide (Aerobid®), Afviar®, Symbicort®, and Dulera®, cromolyn sodium (Intal®), methylxanthines such as theophylline (Theo-Dur®, Theolair®, Slo-bid®, Uniphyl®, Theo-24®) and aminophylline, and IgE antibodies such as omalizumab (Xolair®).

[0251] 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 (Alupent®), pirbuterol acetate (Maxair®), terbutaline sulfate (Brethaire®), salmeterol xinafoate (Serevent®), and formoterol (Foradil®), an anticholinergic such as ipratropium bromide (Atrovent®) and tiotropium (Spiriva®), a methylxanthine such as theophylline (Theo- and administering one or more additional therapeutic agents selected from inhaled corticosteroids such as Dur®, Theolair®, Slo-bid®, Uniphyl®, Theo-24®, and aminophylline, including prednisone, prednisolone, beclomethasone dipropionate (Beclovent®, Qvar®, and Vanceril®), triamcinolone acetonide (Azmacort®), mometasone (Asthmanex®), budesonide (Pulmocort®), flunisolide (Aerobid®), Afviar®, Symbicort®, and Dulera®.

[0252] In another embodiment, the present 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.

[0253] In another embodiment, the present 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.

[0254] In another embodiment, the present 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).

[0255] In another embodiment, the present 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.

[0256] In another embodiment, the present invention provides a method of treating multiple myeloma, comprising administering to a patient in need thereof the 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®).

[0257] 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 method comprises administering to the patient a compound of Formula I and a BTK inhibitor. In another embodiment, the present invention provides a method of treating or lessening the severity of a disease, comprising administering to the patient a compound of Formula I and a BTK inhibitor, wherein the method comprises administering to the patient a compound of Formula I and a BTK inhibitor. In another embodiment, the present invention provides a method of treating or lessening the severity of a disease, comprising administering to the patient a compound of Formula I and a BTK inhibitor, wherein the method comprises administering to the patient a compound of Formula I and a BTK inhibitor. In another embodiment, the present invention provides a method of treating or lessening the severity of a disease, comprising administering to the patient a compound of Formula I and a BTK inhibitor, spondylosis), antiphospholipid 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, autonomic neuropathy, membranous glomerulonephritis, endometriosis, interstitial cystitis, pemphigus vulgaris, bullous pemphigoid, neuromyotonia, scleroderma, vulvodynia, hyperproliferative disorders, rejection after organ or tissue transplantation, acquired immune deficiency syndrome (AIDS, also known as HIV), type 1 diabetes, graft-versus-host disease, transplantation, blood transfusion, anaphylaxis, allergies Allergies (e.g., allergies to plant pollen, latex, drugs, foods, insect venom, animal hair, animal dander, dust mites, or the calyx of the mushroom body of a cockroach), type I hypersensitivity, allergic conjunctivitis, allergic rhinitis, and atopic dermatitis, asthma, appendicitis, atopic dermatitis, asthma, allergies, blepharitis, bronchiolitis, bronchitis, bursitis, uterus Cervicitis, cholangitis, cholecystitis, chronic graft rejection, colitis, conjunctivitis, Crohn's disease, cystitis, dacryoadenitis, dermatitis, dermatomyositis, encephalitis, endocarditis, endometritis, enteritis, enterocolitis, epicondylitis, epididymitis, fasciitis, fibrositis, 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, e.g., diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic lymphoma, chronic lymphocytic leukemia, acute lymphocytic leukemia, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma / Waldenstrom's macroglobulinemia, splenic marginal zone lymphoma, multiple myeloma (plasmocyte myeloma), follicular 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 Bone and joint diseases, including ankylosing spondylitis, psoriatic arthritis, and Reiter's disease, Behcet's disease, Sjögren's syndrome, systemic sclerosis, osteoporosis, bone cancer, bone metastases, thromboembolic disorders (e.g., myocardial infarction, angina pectoris, re-occlusion after angioplasty, restenosis after angioplasty, re-occlusion after aortocoronary artery bypass grafting, restenosis after aortocoronary artery bypass grafting, stroke, transient ischemia, peripheral arterial occlusive disease, pulmonary embolism, deep vein thrombosis), inflammatory pelvic disease, urethritis, sunburn, sinusitis, pneumonia, encephalitis, meningitis, myocarditis, nephritis, osteomyelitis, myositis, hepatitis, gastritis, enteritis, dermatitis , gingivitis, appendicitis, pancreatitis, cholecystitis (cholocystitus), agammaglobulinemia, psoriasis, allergies, Crohn's disease, irritable bowel syndrome, ulcerative colitis, Sjogren's syndrome, tissue graft rejection, hyperacute rejection after organ transplant, 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,Selected from Addison's disease, Parkinson's disease, Alzheimer's disease, diabetes, septic shock, cutaneous lupus erythematosus, systemic lupus erythematosus (SLE), rheumatoid arthritis, psoriatic arthritis, juvenile arthritis, osteoarthritis, chronic idiopathic thrombocytopenic purpura, Waldenstrom's macroglobulinemia, myasthenia gravis, Hashimoto's disease, atopic dermatitis, osteoarthritis, 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.

[0258] 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 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.

[0259] 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, including carcinoma or solid tumor of the brain, kidney (e.g., renal cell carcinoma (RCC)), liver, adrenal gland, bladder, breast, stomach, gastric tumor, ovarian, colon, rectum, prostate, pancreas, lung, vagina, endometrium, cervix, testis, genitourinary tract, esophagus, larynx, skin, bone, or thyroid, sarcoma, glioblastoma, neuroblastoma, multiple bone marrow tumor, or thyroid cancer. myeloma, or gastrointestinal cancer, in particular colon cancer or colorectal adenoma or tumors of the head and neck, 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) and Hodgkin's lymphoma (also called Hodgkin's lymphoma or Hodgkin's disease)), breast cancer, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, or leukemia, e.g., Cowden's syndrome, Lhermitte-Duclos diseaseasthma of any type or genesis, such as acute, arachidic, catarrhal, croupus, bronchial asthma, chronic obstructive pulmonary disease, and diseases including Bannayan-Zonana syndrome, or diseases in which the PI3K / PKB pathway is aberrantly activated; asthma of any type or genesis, such as acute, arachidic, catarrhal, croupus, bronchial asthma, chronic obstructive pulmonary disease, chronic obstructive pulmonary disease, chronic obstructive pulmonary disease, acute lung injury (ALI), acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease, airway or lung disease (COPD, COAD, or COLD), such as chronic bronchitis or related dyspnea, emphysema, and asthma including exacerbation of airway hyperresponsiveness as a result of other medications, especially other inhaled medications; bronchitis of any kind or genesis, including but not limited to bronchitis, chronic, or tuberculous-like bronchitis; pneumoconiosis of any kind or genesis (an inflammatory, usually 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 pneumonia, ostrich ptilosis, siderosis, silicosis, tobacco pneumonitis, 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 multiformemultiforme, 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 associated with or has an autoimmune component or etiology, including autoimmune blood disorders (e.g., hemolytic anemia, aplastic anemia, pure red cell aplasia, and idiopathic thrombocytopenia), cutaneous lupus erythematosus, systemic lupus erythematosus, rheumatoid arthritis, polychondritis, scleroderma, Wegener's granulomatosis granulamatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, Stevens-Johnson syndrome, idiopathic sprue, autoimmune inflammatory bowel disease (e.g., ulcerative colitis and Crohn's disease), endocrine opthalmopathy, Graves' disease, sarcoidosis, alveolitis, chronic hypersensitivity pneumonitis, multiple sclerosis, primary biliary cirrhosis, uveitis (anterior and posterior), keratoconjunctivitis sicca and vernal keratoconjunctivitis, interstitial pulmonary fibrosis, psoriatic arthritis, and glomerulonephritis (with and without nephrotic syndrome, e.g., idiopathic nephrotic syndrome or minimal change nephrosis) The disease is selected from neurodegenerative diseases caused by traumatic injury, glutamate neurotoxicity, and hypoxia, including cerebrovascular disease (including cerebrovascular accident), 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, as well as neurodegenerative diseases caused by traumatic injury, glutamate neurotoxicity, and hypoxia.

[0260] In some embodiments, 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 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 disease, small cell lung cancer, non-small cell lung cancer, myelodysplastic syndrome, lymphoma, a hematological tumor, or a solid tumor.

[0261] 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 WO2014074660A1, WO2014074661A1, and WO2015089143A1, each of which is incorporated by reference in its entirety. Suitable JH1 domain binding compounds include those described in WO2015131080A1, each of which is incorporated by reference in its entirety.

[0262] The compounds and compositions of the present invention can be administered at any dosage and via any route effective for treating or reducing 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 specific 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 dosage 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 being treated and the severity of the disorder, the activity of the particular compound used, the particular composition used, the patient's age, weight, general health, sex, and diet, the time of administration, route of administration, and excretion rate of the particular compound used, the duration of treatment, drugs used in conjunction with or concurrently with the particular compound used, and similar factors well known in the medical arts. As used herein, the term "patient" means an animal, preferably a mammal, and most preferably a human.

[0263] The pharmaceutically acceptable compositions of the present invention can be administered to humans and other animals orally, rectally, parenterally, intravesically, intravaginally, intraperitoneally, topically (as powders, ointments, 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 can 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.

[0264] 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, 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 sesame oil), emulsifiers such as glycerin, 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.

[0265] 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.

[0266] 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.

[0267] 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 dissolution rate, which may in turn depend on crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form can be achieved by dissolving or suspending the compound in an oil vehicle. Depot injectable formulations 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 used, 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.

[0268] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing a compound of the present invention with a suitable non-irritating excipient or carrier such as cocoa butter, polyethylene glycol, or a suppository wax, which are solid at ambient temperature but liquid at body temperature and will melt in the rectum or vaginal cavity and release the active compound.

[0269] 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 glycerin; 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.

[0270] 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 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. 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.

[0271] 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.

[0272] Dosage forms for topical or transdermal administration of the compounds of the present invention include ointments, pastes, emulsions, 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 any, that may be required. Ophthalmic formulations, ear drops, and eye drops are also contemplated within the scope of the present invention. Additionally, 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 either providing a rate-controlling membrane or dispersing the compound in a polymer matrix or gel.

[0273] 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.

[0274] 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 a particular embodiment, the present invention relates to a method for irreversibly 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.

[0275] 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.

[0276] 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.

[0277] 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.

[0278] According to another embodiment, the present invention relates to a method for 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 for reversibly or irreversibly inhibiting one or more activities 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. 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.

[0279] Depending on the particular condition, or disease, to be 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, to be treated."

[0280] 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 anti-angiogenic 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 function modifiers; antiproliferative antibodies; heparanase inhibitors; inhibitors of oncogenic Ras isoforms; telomerase inhibitors; proteasome inhibitors; compounds used in the treatment of hematological malignancies; compounds that target, reduce or inhibit the activity of Flt-3; e.g. 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; temozolomide (Temodal®); kinesin spindle protein inhibitors such as, for example, SB715992 or SB743921 from GlaxoSmithKline, or pentamidine / chlorpromazine from CombinatoRx; MEK inhibitors such as, for example, ARRY142886 from Array BioPharma, AZD6244 from AstraZeneca, PD181461 from Pfizer, and leucovorin.As used herein, the term "aromatase inhibitor" refers to a compound that inhibits estrogen production, for example, the conversion of the substrates androstenedione and testosterone to estrone and estradiol, respectively. This term includes, but is not limited to, steroids, particularly atamestane, exemestane, and formestane, as well as 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 invention that include a chemotherapeutic agent that is an aromatase inhibitor are particularly useful in the treatment of hormone receptor positive tumors, such as breast tumors.

[0281] As used herein, the term "antiestrogen" 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 combinations of the present invention that include chemotherapeutic agents that are antiestrogens are particularly useful for treating estrogen receptor-positive tumors, such as breast tumors.

[0282] As used herein, the term "antiandrogen" refers to any substance capable of inhibiting the biological effects of male 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™.

[0283] As used herein, the term "topoisomerase I inhibitor" includes, but is not limited to, topotecan, gimatecan, irinotecan, camptothecian and its analogs, 9-nitrocamptothecin, and the polymeric camptothecin conjugate PNU-166148. Irinotecan can be administered, for example, in the form as marketed, for example, under the trademark Camptosar™. Topotecan is sold under the trade name Hycamptin™.

[0284] As used herein, the term "topoisomerase II inhibitor" includes, but is not limited to, doxorubicin (including liposomal formulations such as Caelyx™), anthracyclines such as daunorubicin, epirubicin, idarubicin, and nemorubicin, the anthraquinones mitoxantrone and losoxantrone, and the podophyllotoxines etoposide and teniposide. Etoposide is sold under the trade name Etopophos™. Teniposide is sold under the trade name VM26-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.

[0285] 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™.

[0286] 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™.

[0287] The term "histone deacetylase inhibitors" or "HDAC inhibitors" relates to compounds which inhibit histone deacetylase and which have antiproliferative activity, including, but not limited to, suberoylanilide hydroxamic acid (SAHA).

[0288] The term "antineoplastic antimetabolite" includes, but is not limited to, 5-fluorouracil or 5-FU, capecitabine, gemcitabine, DNA demethylating compounds such as 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™.

[0289] As used herein, the term "platin compound" includes, but is not limited to, carboplatin, cis-platin, cisplatinum, and oxaliplatin. Carboplatin can be administered, e.g., in the form as it is marketed, e.g., under the trademark Carboplat™. Oxaliplatin can be administered, e.g., in the form as it is marketed, e.g., under the trademark Eloxatin™.

[0290] As used herein, "compounds that target / reduce protein or lipid kinase activity, or protein or lipid phosphatase activity;The term "or further anti-angiogenic compounds" refers to, for example, a) compounds which target, decrease or inhibit the activity of platelet-derived growth factor receptors (PDGFR), such as compounds which target, decrease or inhibit the activity of PDGFR, in particular compounds which inhibit PDGF receptors, such as N-phenyl-2-pyrimidine-amine derivatives, such as imatinib, SU101, SU6668, and GFB-111; b) compounds which target, decrease or inhibit the activity of fibroblast growth factor receptors (FGFR). c) compounds that target, decrease or inhibit the activity of insulin-like growth factor receptor I (IGF-IR), such as compounds that target, decrease or inhibit the activity of IGF-IR, in particular 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, decrease or inhibit the activity of the Trk receptor tyrosine kinase family or ephrin B4 inhibitors; e) AxI receptor tyrosine kinase f) compounds that target, reduce or inhibit the activity of the Ret receptor tyrosine kinase; g) compounds that target, reduce or inhibit the activity of the Kit / SCFR receptor tyrosine kinase, e.g., imatinib; h) compounds that target, reduce or inhibit the activity of the C-kit receptor tyrosine kinase, which is part of the PDGFR family, e.g., compounds that target, reduce or inhibit the activity of the c-Kit receptor tyrosine kinase family. i) compounds that inhibit the activity of c-Abl family members, their gene fusion products (e.g., BCR-Abl kinase) and mutants, such as N-phenyl-2-pyrimidine-amine derivatives, e.g., imatinib or nilotinib (AMN107); PD180970; AG957; NSC 680410;PD173955 from ParkeDavis; or dasatinib (BMS-354825), j) compounds which 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 exemplary compounds include UCN-01, safingol, BAY 43-9006, bryostatin 1, perifosine; ilmofosine; RO 318220 and RO 320432; GO 6976; Isis 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, including 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 adaphostin (4-{[(2,5-dihydroxyphenyl)methyl]amino}-benzoic acid adamantyl ester;NSC 680410, adaphostin), l) compounds which target, decrease or inhibit the activity of the epidermal growth factor family of receptor tyrosine kinases (EGFR1 ErbB2, ErbB3, ErbB4 as homodimers or heterodimers) and their variants, for example compounds which target, decrease or inhibit the activity of the epidermal growth factor receptor family, in particular compounds, proteins or antibodies which inhibit members of the EGF receptor tyrosine kinase family, such as EGF receptor, ErbB2, ErbB3 and ErbB4, or which bind to EGF or EGF-related ligands, CP 358774, ZD 1839, ZM 105180;m) compounds that target, decrease or inhibit the activity of c-Met, particularly the c-Met receptor, 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. compounds that target, decrease, or inhibit the activity of the c-Met receptor, such as compounds that inhibit kinase activity of the c-Met receptor or antibodies that target the extracellular domain of c-Met or that bind to HGF; n) compounds that inhibit one or more JAK family members (JAK1 / JAK2 / JAK3 / TYK2 and / or pan-JAK), including, but not limited to, 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 PI3 kinase (PI3K), including but not limited to ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, buparlisib, pictrelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765, and idelalisib. and 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 signaling effects of the Hedgehog (Hh) or Smoothened receptor (SMO) pathways, including but not limited to cyclopamine, vismodegib, itraconazole, erismodegib, and IPI-926 (saridegib);

[0291] 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, but are not limited to, ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, buparlisib, pictrelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765, and idelalisib.

[0292] 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.

[0293] 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.

[0294] As used herein, the term "Bcl-2 inhibitor" includes compounds such as ABT-199, ABT-731, ABT-737, apogossypol, Ascenta's pan-Bcl-2 inhibitors, curcumin (and analogs thereof), dual Bcl-2 / Bcl-xL inhibitors (Infinity Pharmaceuticals / Novartis Pharmaceuticals), Genasense (G3139), HA14-1 (and analogs thereof, see WO 2008118802), navitoclax (and analogs thereof, see U.S. Pat. No. 7,390,799), NH-1 (Shenayng Pharmaceutical University), obatoclax (and analogs thereof, see WO 2004106328), S-001 (Gloria Pharmaceuticals), TW series compounds (Univ. of Michigan), and others. These include, but are not limited to, compounds that have inhibitory activity against B-cell lymphoma 2 protein (Bcl-2), including, but not limited to, venetoclax, and venetoclax. In some embodiments, the Bcl-2 inhibitor is a small molecule therapeutic. In some embodiments, the Bcl-2 inhibitor is a peptidomimetic.

[0295] Further examples of BTK inhibitory compounds and conditions treatable by such compounds in combination with the compounds of the invention can be found in WO2008039218 and WO2011090760, which are incorporated by reference in their entireties.

[0296] 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 WO2003063794, WO2005007623, and WO2006078846, which are incorporated by reference in their entireties.

[0297] 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 WO2004019973, WO2004089925, WO2007016176, U.S. Pat. No. 8,138,347, WO2002088112, WO2007084786, WO2007129161, WO2006122806, WO2005113554, and WO2007044729, which are incorporated herein by reference in their entireties.

[0298] 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 WO2009114512, WO2008109943, WO2007053452, WO2000142246, and WO2007070514, which are incorporated by reference in their entireties.

[0299] Additional antiangiogenic compounds include, for example, compounds that have another mechanism for their activity that is not related to protein or lipid kinase inhibition, such as thalidomide (Thalomid™) and TNP-470.

[0300] Examples of useful proteasome inhibitors for use 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.

[0301] 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.

[0302] 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.

[0303] As used herein, the term cyclooxygenase inhibitor 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.

[0304] 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. Etridonic 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 have antiproliferative activity, such as sirolimus (Rapamune®), everolimus (Certican™), CCI-779 and ABT578.

[0305] 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 function modifier" refers to a lymphokine or interferon.

[0306] As used herein, the term "inhibitor of an oncogenic Ras isoform," such as H-Ras, K-Ras, or N-Ras, refers to a compound that targets, reduces, or inhibits the oncogenic activity of Ras, e.g., a "farnesyltransferase inhibitor" such as L-744832, DK8G557, or R115777 (Zarnestra™). As used herein, the term "telomerase inhibitor" refers to a compound that targets, reduces, or inhibits 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.

[0307] 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.

[0308] 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.

[0309] 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.

[0310] 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.

[0311] 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.

[0312] 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 compounds that degrade, target, reduce, or inhibit 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.

[0313] 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. By antibody is meant 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.

[0314] In the treatment of acute myeloid leukemia (AML), the compounds of the present invention can be used in combination with standard leukemia therapies, particularly in combination with therapies used to treat AML. In particular, the compounds of the present invention can be administered in combination with farnesyltransferase inhibitors and / or other agents useful in 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, lestaurtinib, sorafenib, and sunitinib.

[0315] 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 U.S. 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-hydroxyethyl){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).

[0316] 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.

[0317] Also, particularly, such compounds, proteins, or monoclonal antibodies against VEGF include, for example, 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine or a pharmaceutically acceptable salt thereof, 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine succinate; angiostatin (Angiostatin™); endostatin (Endostatin™); anthranilic acid amide; ZD4190; ZD6474; SU5416; SU6668; bevacizumab; or anti-VEGF antibodies or anti-VEGF receptor antibodies, for example, rhuMAb and RHUFab, VEGF aptamers, for example, Macugon; FLT-4 inhibitors, FLT-3 inhibitors, VEGFR-2 IgGI antibodies, Angiozyme (RPI), 4610), and bevacizumab (Avastin™).

[0318] 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.

[0319] As used herein, angiogenic antisteroids refer to compounds that block or inhibit angiogenesis, such as anecortave, triamcinolone, hydrocortisone, 11-α-epihydrocotisol, cortexolone, 17α-hydroxyprogesterone, corticosterone, desoxycorticosterone, testosterone, estrone, and dexamethasone.

[0320] Placement agents containing corticosteroids refer to compounds such as fluocinolone and dexamethasone.

[0321] Other chemotherapeutic compounds include, but are not limited to, plant alkaloids, hormonal compounds and antagonists; biological function 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.

[0322] 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, bronchodilator, 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 necessary dosing or potential side effects of such drugs. The compounds of the present invention can be mixed with the other drug substances in a given 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, bronchodilator, antihistamine, or antitussive drug substances, wherein the compounds of the present invention and the drug substances are present in the same or different pharmaceutical compositions.

[0323] Suitable anti-inflammatory drugs include steroids, particularly 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), Arofylline (Almirall Prodesfarma), PD189659 / PD168787 (Parke-Davis), AWD-12-281 (Asta Medica), CDC-801 (Celgene), SeICID (trademark), CC-10004 (Celgene), VM554 / UM565 (Vernalis), T-440 (Tanabe), KW-4490 (Kyowa Hakko Kogyo) and beta-2 adrenoceptor 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.

[0324] 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.

[0325] Other useful combinations of compounds of the invention with anti-inflammatory agents are 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 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).

[0326] The structures of active compounds identified by code number, generic name or trade name can be obtained from the actual edition of the standard catalogue "The Merck Index" or from databases such as international patents (e.g. IMS World Publications).

[0327] Exemplary Cancer Immunotherapeutic Agents In some embodiments, the one or more other therapeutic agents are cancer immunotherapeutic agents. As used herein, the term "immuno-oncology agent" refers to an agent that is effective in enhancing, stimulating, and / or upregulating an immune response in a subject. In some embodiments, administration of cancer immunotherapeutic agents, including compounds of the present invention, has a synergistic effect in the treatment of cancer.

[0328] The cancer immunotherapeutic agent can be, for example, a small molecule drug, an antibody, or a biological molecule or small molecule. Examples of biological cancer immunotherapeutics 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.

[0329] In some embodiments, the cancer immunotherapeutic 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.

[0330] Certain stimulatory and inhibitory molecules are members of the immunoglobulin superfamily (IgSF). One important family of membrane-bound ligands that bind to costimulatory or costimulatory 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 costimulatory 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, RANK, 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.

[0331] In some embodiments, the cancer immunotherapeutic 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.

[0332] In some embodiments, the combination of a compound of the invention with a cancer immunotherapeutic agent can stimulate a T cell response. In some embodiments, the cancer immunotherapeutic 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.

[0333] In some embodiments, the cancer immunotherapeutic 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 cancer immunotherapeutic agent is an antagonist of a KIR, such as lirilumab.

[0334] In some embodiments, the cancer immunotherapeutic agent is an agent that inhibits or depletes macrophages or monocytes, including but not limited to CSF-1R antagonists such as RG7155 (WO 11 / 70024, WO 11 / 107553, WO 11 / 131407, WO 13 / 87699, WO 13 / 119716, WO 13 / 132044), or CSF-1R antagonist antibodies, including FPA-008 (WO 11 / 140249, WO 13169264, WO 14 / 036357).

[0335] In some embodiments, the cancer immunotherapeutic 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 (regulatory T cells) (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 activate innate immunity and / or induce inflammation at the tumor site.

[0336] In some embodiments, the cancer immunotherapeutic agent is a CTLA-4 antagonist. In some embodiments, the CTLA-4 antagonist is a CTLA-4 antagonist antibody. In some embodiments, the CTLA-4 antagonist antibody is YERVOY (ipilimumab) or tremelimumab.

[0337] In some embodiments, the cancer immunotherapeutic agent is a PD-1 antagonist. In some embodiments, the PD-1 antagonist is administered by infusion. In some embodiments, the cancer immunotherapeutic agent is an antibody or antigen-binding portion thereof that specifically binds to the programmed cell death-1 (PD-1) receptor and inhibits PD-1 activity. In some embodiments, the PD-1 antagonist is a PD-1 antagonist antibody. In some embodiments, the PD-1 antagonist antibody is OPDIVO (nivolumab), KEYTRUDA (pembrolizumab), or MEDI-0680 (AMP-514; WO 2012 / 145493). In some embodiments, the cancer immunotherapeutic agent can be pidilizumab (CT-011). In some embodiments, the cancer immunotherapeutic 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.

[0338] In some embodiments, the cancer immunotherapeutic agent is a PD-L1 antagonist. In some embodiments, the PD-L1 antagonist is a PD-L1 antagonist antibody. In some embodiments, the PD-L1 antibody is MPDL3280A (RG7446; WO 2010 / 077634), durvalumab (MEDI4736), BMS-936559 (WO 2007 / 005874), and MSB0010718C (WO 2013 / 79174).

[0339] In some embodiments, the cancer immunotherapeutic agent is a LAG-3 antagonist. In some embodiments, the LAG-3 antagonist is a LAG-3 antagonist antibody. In some embodiments, the LAG3 antibody is BMS-986016 (WO 10 / 19570, WO 14 / 08218), or IMP-731 or IMP-321 (WO 8 / 132601, WO 009 / 44273).

[0340] In some embodiments, the cancer immunotherapeutic agent is a CD137 (4-1BB) agonist. In some embodiments, the CD137 (4-1BB) agonist is a CD137 agonist antibody. In some embodiments, the CD137 antibody is urelumab or PF-05082566 (WO 12 / 32433).

[0341] In some embodiments, the cancer immunotherapeutic agent is a GITR agonist. In some embodiments, the GITR agonist is a GITR agonist antibody. In some embodiments, the GITR antibody is BMS-986153, BMS-986156, TRX-518 (WO 006 / 105021, WO 009 / 009116), or MK-4166 (WO 11 / 028683).

[0342] In some embodiments, the cancer immunotherapeutic agent is an indoleamine (2,3)-dioxygenase (IDO) antagonist. In some embodiments, the IDO antagonist is selected from the group consisting of 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), and kynurenine-degrading enzymes (Kynase, Ikena). Oncology, Inc., formerly known as Kyn Therapy), and NLG-919 (WO 09 / 73620, WO 009 / 1156652, WO 11 / 56652, WO 12 / 142237).

[0343] In some embodiments, the cancer immunotherapeutic agent is an OX40 agonist. In some embodiments, the OX40 agonist is an OX40 agonist antibody. In some embodiments, the OX40 antibody is MEDI-6383 or MEDI-6469.

[0344] In some embodiments, the cancer immunotherapeutic agent is an OX40L antagonist. In some embodiments, the OX40L antagonist is an OX40 antagonist antibody. In some embodiments, the OX40L antagonist is RG-7888 (WO 06 / 029879).

[0345] In some embodiments, the cancer immunotherapeutic is a CD40 agonist. In some embodiments, the CD40 agonist is a CD40 agonist antibody. In some embodiments, the cancer immunotherapeutic is a CD40 antagonist. In some embodiments, the CD40 antagonist is a CD40 antagonist antibody. In some embodiments, the CD40 antibody is lucatumumab or dacetuzumab.

[0346] In some embodiments, the cancer immunotherapeutic agent is a CD27 agonist. In some embodiments, the CD27 agonist is a CD27 agonist antibody. In some embodiments, the CD27 antibody is varlilumab.

[0347] In some embodiments, the cancer immunotherapeutic is MGA271 (directed against B7H3) (WO 11 / 109400).

[0348] In some embodiments, the cancer immunotherapeutic agent is abagovomab, adecatumumab, afutuzumab, alemtuzumab, anatumomab mafenatox, apolizumab, atezolimab, avelumab, blinatumomab, BMS-936559, catumaxomab, durvalumab, epacadostat, epratuzumab, indoximod, inotuzumab ozogamicin, intelumumab, ipilimumab, isatuximab, ran brolizumab, MED14736, MPDL3280A, nivolumab, obinutuzumab, ocaratuzumab, ofatumumab, olatatumab, pembrolizumab, pidilizumab, rituximab, ticilimumab, samalizumab, or tremelimumab.

[0349] In some embodiments, the cancer immunotherapeutic 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, which have been shown in clinical trials to induce durable anti-tumor responses in an increasing number of tumor histologies, including several 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.

[0350] In some embodiments, the immunomodulatory therapeutic agent specifically induces apoptosis in 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).

[0351] In some embodiments, the cancer immunotherapeutic agent is a cancer vaccine, hi 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 lesions of melanoma. In some embodiments, the cancer immunotherapeutic agent is an oncolytic virotherapy, such as pexastimogene devacirepvec (PexaVec / JX-594, SillaJen Inc. / formerly Jennerex), a thymidine kinase (TK)-deficient vaccinia virus engineered to express GM-CSF for hepatocellular carcinoma (NCT02562755) and melanoma (NCT00429312). Oncolytics Biotherapeutics; REOLYSIN® (Oncolytics), a variant of the respiratory enteric orphan virus (reovirus) that does not replicate in cells that are not RAS-activated, 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; 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 ovarian cancer (NCT02028117), metastatic or advanced epithelial tumors such as colorectal cancer, bladder cancer, head and neck squamous cell carcinoma, and salivary gland cancer (NCT02636036);ONCOS-102 (Targovax / formerly Oncos), an adenovirus engineered to express GM-CSF in melanoma (NCT03003676), and peritoneal disease, colorectal cancer, or ovarian cancer (NCT02963831); GL-ONC1 (GLV-1h68 / GLV-1h153, Genelux GmbH), a vaccinia virus engineered to express beta-galactosidase (beta-gal) / beta-glucoronidase or beta-gal / human sodium iodide symporter (hNIS), respectively, which has been tested in peritoneal carcinomatosis (NCT01443260), fallopian tube cancer, and ovarian cancer (NCT02759588); or CG0070 (Cold Genesys), an adenovirus engineered to express GM-CSF in bladder cancer (NCT02365818). ;

[0352] In some embodiments, the cancer immunotherapeutics include JX-929 (SillaJen / formerly Jennerex Biotherapeutics), a TK and vaccinia growth factor-deficient vaccinia virus engineered to express cytosine deaminase, which can convert 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; and TILT-123 (TILT Biotherapeutics), an engineered adenovirus called Ad5 / 3-E2F-delta24-hTNFα-IRES-hIL20; and 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.

[0353] In some embodiments, the cancer immunotherapeutic 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.

[0354] CARs have been constructed that consist of a binding domain that can be 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.

[0355] 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, CARs 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]

[0356] 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 (NCT02929862) for the treatment of solid tumors.

[0357] In some embodiments, the immunostimulatory agent is an agonist or activator of a toll-like receptor (TLR). Suitable activators of TLRs include agonists or activators of TLR9, such as SD-101 (Dynavax). SD-101 is an immunostimulatory CpG that is being tested for B-cell, follicular lymphoma, and other lymphomas (NCT02254772). Agonists or activators of TLR8 that can be used in the present invention include motolimod (VTX-2337, VentiRx Pharmaceuticals), which is being tested for squamous cell carcinoma of the head and neck (NCT02124850) and ovarian cancer (NCT02431559).

[0358] Other cancer immunotherapeutic agents that can be used in the present invention include urelumab (BMS-663513, Bristol-Myers Squibb), an anti-CD137 monoclonal antibody; varlilumab (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, Inc.,). Anti-MMP9 antibody; MK-4166 (Merck & Co.), anti-GITR monoclonal antibody.

[0359] In some embodiments, the immunostimulatory agent is selected from elotuzumab, mifamurtide, agonists or activators of toll-like receptors, and activators of RORγt.

[0360] In some embodiments, the immune stimulatory therapy is recombinant human interleukin-15 (rhIL-15). rhIL-15 has been 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 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.

[0361] In some embodiments, the cancer immunotherapeutic 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 cancer immunotherapeutic agent is selected from the examples described in Table 1 of Jerry L. Adams et al. In some embodiments, the cancer immunotherapeutic agent is a small molecule that targets a cancer immunotherapy target selected from those listed in Table 2 of Jerry L. Adams et al. In some embodiments, the cancer immunotherapeutic agent is a small molecule agent selected from those listed in Table 2 of Jerry L. Adams et al.

[0362] In some embodiments, the cancer immunotherapeutic agent is selected from the small molecule cancer immunotherapeutic 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 cancer immunotherapeutic agent is an agent that targets a pathway described in Peter L. Toogood.

[0363] In some embodiments, the cancer immunotherapeutic 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 cancer immunotherapeutic 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, which 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, which results in the induction of bystander cell lysis. In some embodiments, the bystander cells are in 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 cancer immunotherapeutic is an antibody that blocks the PD-L1 / PD1 axis and / or CTLA4. In some embodiments, the cancer immunotherapeutic is ex vivo expanded tumor-infiltrating T cells. In some embodiments, the cancer immunotherapeutic is a bispecific antibody construct or a chimeric antigen receptor (CAR) that directly links T cells to tumor-associated surface antigens (TAA).

[0364] Exemplary Immune Checkpoint Inhibitors In some embodiments, the cancer immunotherapeutic is an immune checkpoint inhibitor described herein.

[0365] As used herein, the term "checkpoint inhibitor" refers to a drug useful for preventing cancer cells from evading a patient's immune system. One of the main mechanisms of anti-tumor immune destruction is known as "T cell exhaustion," which is caused by the upregulation of inhibitory receptors by chronic exposure to antigens. These inhibitory receptors function as immune checkpoints to prevent uncontrolled immune responses.

[0366] PD-1 and coinhibitory 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.

[0367] 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.

[0368] 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 a further aspect, the vaccine is a dendritic cell (DC) vaccine.

[0369] 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 belongs to the CD2 family of molecules and is involved in NK, gamma delta, and memory CD8+ (expressed on all αβ T cells), CD160 (also known as BY55), 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.

[0370] 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).

[0371] 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.

[0372] In some embodiments, the immune checkpoint inhibitor is REGN2810 (Regeneron), an anti-PD-1 antibody that has been 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 to PD-1 in clinical trials in non-small cell lung cancer, Merkel cell carcinoma, mesothelioma, solid tumors, renal cancer, ovarian cancer, bladder cancer, head and neck cancer, and gastric cancer; avelumab (BAVENCIO®, Pfizer / Merck KGaA), also known as MSB0010718C, a fully human IgG1 anti-PD-L1 antibody in clinical trials in non-small cell lung cancer, Merkel cell carcinoma, mesothelioma, solid tumors, renal cancer, ovarian cancer, bladder cancer, head and neck cancer, and gastric cancer; and PDR001 (Novartis), an inhibitory antibody that binds to PD-1 in clinical trials in 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 directed against CTLA-4 and is being studied in clinical trials for multiple 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, liver metastases, 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).

[0373] 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 tested in solid tumors (NCT02817633). LY3321367 (Eli Lilly) is an anti-TIM-3 antibody being tested in solid tumors (NCT03099109). MBG453 (Novartis) is an anti-TIM-3 antibody being tested in advanced malignancies (NCT02608268).

[0374] 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).

[0375] 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 tested in glioblastoma and gliosarcoma (NCT02658981). REGN3767 (Regeneron) is also an anti-LAG-3 antibody and is being tested in malignant tumors (NCT03005782). IMP321 (Immutep SA), an LAG-3-Ig fusion protein, is being tested in melanoma (NCT02676869), adenocarcinoma (NCT02614833), and metastatic breast cancer (NCT00349934).

[0376] Checkpoint inhibitors that can be used in the present invention include OX40 agonists. OX40 agonists being tested in clinical trials include PF-04518600 / PF-8600 (Pfizer), an anti-OX40 agonist antibody, in metastatic renal cancer (NCT03092856) and advanced cancers and neoplasms (NCT02554812; NCT05082566); GSK3174998 (Merck), an anti-OX40 agonist antibody, in a Phase 1 cancer trial (NCT02528357); and GSK3174998 (Merck), an anti-OX40 agonist antibody, in advanced solid tumors (NCT02318394 and NCT02705482). These include MEDI0562 (Medimmune / AstraZeneca); MEDI6469, an anti-OX40 agonist 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 anti-OX40 agonist antibody, in advanced cancers (NCT02737475).

[0377] Checkpoint inhibitors that may be used in the present invention include CD137 (also known as 4-1BB) agonists. CD137 agonists being tested in clinical trials include the anti-CD137 agonist antibody utomilumab (PF-05082566, Pfizer) in diffuse large B-cell lymphoma (NCT02951156) and advanced cancers and neoplasms (NCT02554812 and NCT05082566); the anti-CD137 agonist antibody urelumab (BMS-663513, Bristol-Myers Squibb) in melanoma and skin cancer (NCT02652455) and glioblastoma and gliosarcoma (NCT02658981); and the anti-CD137 agonist antibody CTX-471 (Compass Therapeutics) in metastatic or locally advanced malignancies (NCT03881488).

[0378] Checkpoint inhibitors that can be used in the present invention include CD27 agonists. CD27 agonists being tested in clinical trials include the anti-CD27 agonist antibody varlilumab (CDX-1127, Celldex Therapeutics) in head and neck squamous cell carcinoma, ovarian cancer, colorectal cancer, renal cell carcinoma, and glioblastoma (NCT02335918), lymphoma (NCT01460134), and glioma and astrocytoma (NCT02924038).

[0379] Checkpoint inhibitors that may be used in the present invention include glucocorticoid-induced tumor necrosis factor receptor (GITR) agonists. GITR agonists being tested in clinical trials include the anti-GITR agonist antibody TRX518 (Leap Therapeutics) in melanoma and other malignant solid tumors (NCT01239134 and NCT02628574); the anti-GITR agonist antibody GWN323 (Novartis) in solid tumors and lymphomas (NCT 02740270); the anti-GITR agonist antibody INCAGN01876 (Incyte / Agenus) in advanced cancers (NCT02697591 and NCT03126110); the anti-GITR agonist antibody MK-4166 (Merck) in solid tumors (NCT02132754); and the human IgG1 This includes MEDI1873 (Medimmune / AstraZeneca), an agonistic hexameric GITR ligand molecule containing an Fc domain.

[0380] Checkpoint inhibitors that can be used in the present invention include inducible T-cell costimulator (ICOS, also known as CD278) agonists. ICOS agonists being tested in clinical trials include MEDI-570 (Medimmune), an anti-ICOS agonist antibody (NCT02520791) in lymphoma; GSK3359609 (Merck), an anti-ICOS agonist antibody, in Phase 1 (NCT02723955); and JTX-2011 (Jounce Therapeutics), an anti-ICOS agonist antibody, in Phase 1 (NCT02904226).

[0381] Checkpoint inhibitors that may be used in the present invention include killer IgG-like receptor (KIR) inhibitors. KIR inhibitors being tested in clinical trials include the anti-KIR antibody lirilumab (IPH2102 / BMS-986015, Innate Pharma / Bristol-Myers Squibb) 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 three domains of the long cytoplasmic tail (KIR3DL2), in lymphoma (NCT02593045).

[0382] 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 tested in clinical trials include ALX-148 (Alexo Therapeutics), an antagonist variant of (SIRPa) that binds to CD47 and inhibits CD47 / SIRPa-mediated signaling, in Phase 1 clinical trials (NCT03013218); TTI-621 (SIRPa-Fc, Trillium), a soluble recombinant fusion protein created by linking the N-terminal CD47-binding domain of SIRPa to the Fc domain of human IgG1, which acts by binding to human CD47 and preventing it from delivering a "no phagocytosis" signal to macrophages, in Phase 1 clinical trials (NCT02890368 and NCT02663518). Therapeutics, Inc.); CC-90002, an anti-CD47 antibody (Celgene) in leukemia (NCT02641002); and Hu5F9-G4 (Forty Seven, Inc.) in colorectal neoplasia and solid tumors (NCT02953782), acute myeloid leukemia (NCT02678338), and lymphoma (NCT02953509).

[0383] Checkpoint inhibitors that can be used in the present invention include CD73 inhibitors. CD73 inhibitors being tested 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).

[0384] 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 tested in clinical trials include MK-1454 (Merck), an agonistic synthetic cyclic dinucleotide, in lymphoma (NCT03010176); and ADU-S100 (MIW815, Aduro Biotech / Novartis), an agonistic synthetic cyclic dinucleotide, in Phase 1 studies (NCT02675439 and NCT03172936).

[0385] Checkpoint inhibitors that can be used in the present invention include CSF1R inhibitors. CSF1R inhibitors being tested in clinical trials include pexidartinib (PLX3397, Plexxikon), a CSF1R small molecule inhibitor, in colorectal cancer, pancreatic cancer, metastatic and advanced cancers (NCT02777710), and melanoma, non-small cell lung cancer, head and neck squamous cell carcinoma, gastrointestinal stromal tumor (GIST), and ovarian cancer (NCT02452424); and pexidartinib (PLX3397, Plexxikon) in pancreatic cancer (NCT03153410), melanoma (NCT03101254), and ovarian cancer (NCT02452424). These include IMC-CS4 (LY3022855, Lilly), an anti-CSF-1R antibody, in advanced 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).

[0386] Checkpoint inhibitors that can be used in the present invention include NKG2A receptor inhibitors. NKG2A receptor inhibitors being tested in clinical trials include monalizumab (IPH2201, Innate Pharma), an anti-NKG2A antibody, in head and neck tumors (NCT02643550) and chronic lymphocytic leukemia (NCT02557516).

[0387] In some embodiments, the immune checkpoint inhibitor is selected from nivolumab, pembrolizumab, ipilimumab, avelumab, durvalumab, atezolizumab, or pidilizumab.

[0388] 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.

[0389] The compounds of the present invention can be administered alone or in combination with one or more other therapeutic compounds, and possible combination therapies take the form of a fixed combination, or the administration of a compound of the present invention and one or more other therapeutic compounds, which are given alternately or independently of each other, 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. Long-term therapy is possible in the context of other treatment strategies, as well as adjuvant therapy, as described above. Other possible treatments are therapy to maintain the patient's condition after tumor regression, or even chemopreventive therapy, for example, in at-risk patients.

[0390] These additional agents can be administered separately from the composition containing the compound of this invention as part of a multiple dose regimen. Alternatively, the agents can be part of a single dosage form, mixed together with the compound of this 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 period of each other, usually within five hours of each other.

[0391] As used herein, the terms "co-administration," "combined," 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.

[0392] The amounts of both the compounds of the present invention and additional therapeutic agent (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 compound of the present invention can be administered.

[0393] 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 dose of 0.01 to 1,000 μg / kg body weight / day of the additional therapeutic agent can be administered.

[0394] The amount of additional therapeutic agent present in the compositions of the present invention will be 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 will be in the range of about 50% to 100% of the amount that would normally be present in a composition comprising that agent as the only therapeutically active agent.

[0395] 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, vascular grafts, 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]

[0396] As shown in the Examples below, in certain exemplary embodiments, compounds are prepared according to the following general procedure. While this general method illustrates the synthesis of certain compounds of the invention, the following general method, and others known to those of skill in the art, can be applied to all compounds, as well as each subclass and species of these compounds, as described herein. Additional compounds of the invention have been prepared by methods substantially similar to those described herein in the Examples and known to those of skill in the art.

[0397] Intermediates Method PB1 - Preparation of 7-fluoro-3-iodoimidazo[1,2-a]pyridine (PB1) [ka] Step-1.7-Fluoroimidazo[1,2-a]pyridine (PB1.1) To a solution of 4-fluoropyridin-2-amine (PB1.0) (50 g, 0.44 mol, 1.0 equiv.) in ethanol (450 ml), chloroacetaldehyde (50% aqueous solution) (300 ml) and sodium bicarbonate (74.9 g, 0.89 mol, 2.0 equiv.) were added, and the reaction was stirred at 60° C. for 4 hours. Upon completion, the reaction mixture was concentrated in vacuo. The resulting crude product was purified by column chromatography, and the compound was eluted with 35% ethyl acetate in hexane to give PB1.1 (50 g, 82.35%), MS (ES): m / z 137.16 [M+H]. + , was obtained.

[0398] Step 2. 7-Fluoro-3-iodoimidazo[1,2-a]pyridine (PB1) To a solution of 7-fluoroimidazo[1,2-a]pyridine (PB1.1) (28 g, 0.20 mol, 1.0 equiv.) in chloroform (300 ml), N-iodosuccinimide (50.65 g, 0.22 mol, 1.1 equiv.) was added and stirred at room temperature for 3 hours. Upon completion, the reaction mixture was quenched with a solution of sodium thiosulfate (1000 ml), extracted with ethyl acetate (600 × 3 ml), dried over sodium sulfate, and concentrated under vacuum. The crude product was purified by column chromatography, and the compound was eluted with 18% ethyl acetate in hexane to give PB1 (30 g, 56.6%), MS (ES): m / z 262.94 [M+H]. + , was obtained.

[0399] Method PB2 - Preparation of 3-iodoimidazo[1,2-a]pyrazine (PB2) [ka] Step 1. Imidazo[1,2-a]pyrazine (PB2.1) To a solution of pyrazin-2-amine (PB2.0) (50 g, 0.52 mol, 1.0 equiv.) in ethanol (450 ml), chloroacetaldehyde (50% aqueous solution) (300 ml, 6 volumes) and sodium bicarbonate (89.2 g, 1.05 mol, 2.0 equiv.) were added, and the reaction was stirred at 60° C. for 4 hours. Upon completion, the reaction mixture was concentrated under vacuum. The resulting crude product was purified by column chromatography, and the compound was eluted with 35% ethyl acetate in hexane to give PB2.1 (50 g, 82.35%), MS (ES): m / z 119.05 [M+H]. + , was obtained.

[0400] Step 2.3-Iodoimidazo[1,2-a]pyrazine (PB2) To a solution of imidazo[1,2-a]pyridine (PB2.1) (28 g, 0.23 mol, 1.0 equiv.) in chloroform (300 ml), N-iodosuccinimide (58.65 g, 0.26 mol, 1.1 equiv.) was added and stirred at room temperature for 3 hours. After completion, the reaction mixture was quenched with a solution of sodium thiosulfate (1000 ml), extracted with ethyl acetate (600 ml x 3), dried over sodium sulfate, and concentrated under vacuum. The crude product was purified by column chromatography, and the compound was eluted with 18% ethyl acetate in hexane to give PB2 (30 g, 56.6%), MS (ES): m / z 245.94 [M+H]. + , was obtained.

[0401] Method PB3—Preparation of methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine (PB3) [ka] Step 1. 4-Bromo-1-methyl-1H-pyrrolo[2,3-b]pyridine (PB3.1) In a three-neck flask, a stirred solution of sodium hydride (60 in mineral oil) (8.12 g, 0.2030 mol, 2.0 equiv.) in N,N-dimethylformamide (100 ml) was treated with a solution of 4-bromo-1H-pyrrolo[2,3-b]pyridine (PB3.0) (20.0 g, 0.0473 mol, 1.0 equiv.) in N,N-dimethylformamide (50 ml), which was added at 0 °C under nitrogen. The reaction was stirred for 30 minutes, and then methyl iodide (7.58 ml, 0.121 mol, 1.2 equiv.) was added and stirred at room temperature for 30 minutes. Upon completion, the reaction mixture was poured into ice-cold water, and the product was extracted with ethyl acetate (300 ml × 3). The organic layers were combined, dried over sodium sulfate, and concentrated under reduced pressure to give crude PB3.1 (20 g, quantitative yield). The crude material was used in the next step without further purification. MS(ES): m / z 211.50[M+H] +

[0402] Step 2. 1-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine (PB3) To a solution of 4-bromo-1-methyl-1H-pyrrolo[2,3-b]pyridine (PB3.1) (10.0 g, 0.047 mol, 1.0 equiv.) and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi-(1,3,2-dioxaborolane) (11.99 g, 0.0473 mol, 1.0 equiv.) in 1,4-dioxane (100 mL), potassium acetate (13.09 g, 0.141 mol, 3.0 equiv.) was added at room temperature. The reaction mixture was degassed with argon gas for 20 minutes, and then [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (3.85 g, 3.85 mol, 0.1 equiv.) was added and stirred at 90 °C for 2.5 hours. After completion, the reaction mixture was poured into water (500 ml) and the product was extracted with ethyl acetate (350 ml x 3). The combined organic layers were washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure to give the crude material. The crude was further purified by column chromatography and the compound was eluted with 15% ethyl acetate in hexane to give PB3 (10.5 g, 82%). m / z 258.5 (M+H) +1

[0403] The following boronic acid intermediates were prepared according to any of the methods PB1-PB3 for the intermediates described above. [Table 4]

[0404] Method PA1 Preparation of 3-(6-amino-2-((dimethylamino)methyl)pyridin-3-yl)cyclopentan-1-ol (PA1) [ka] Step 1. tert-Butyl (5-bromo-6-(hydroxymethyl)pyridin-2-yl)carbamate (PA1.7) A solution of methyl 3-bromo-6-(bis(tert-butoxycarbonyl)amino)picolinate (PA1.6) (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 equiv.) and stirred at 70 °C for 2 h. Upon completion, the ethanol was removed under reduced pressure and water (200 mL) was added dropwise. The crude material was extracted into dichloromethane (3 × 150 mL), and the combined organic layers were washed with brine (100 mL), passed through NaSO, concentrated under reduced pressure, and triturated with diethyl ether (50 mL) to give PA1.7 (27 g, 79%) as a white solid. MS (ES): m / z 395 [M+1] +

[0405] Steps 2 and 3. tert-Butyl (5-bromo-6-((dimethylamino)methyl)pyridin-2-yl)carbamate (PA1.8) A solution of tert-butyl (5-bromo-6-(hydroxymethyl)pyridin-2-yl)carbamate (PA1.7) (22.2 g, 73.2 mmol, 1.0 equiv) and N-N-diisopropylethylamine (33.3 g, 256.3 mmol, 3.5 equiv) in dichloromethane (200 mL) was cooled to 0 °C, treated with methanesulfonyl chloride (12.5 g, 109.8 mmol, 1.5 equiv) and stirred for 30 min. The reaction was quenched with water (100 mL) and extracted with DCM (3 × 40 mL). The combined organic layers were washed with brine, passed through NaSO, and concentrated under reduced pressure. The crude material was dissolved in acetonitrile (200 ml) and to this was added dimethylamine (15 g, 183.0 mmol, 2.5 equiv.) and N,N-diisopropylethylamine (33.3 g, 256.3 mmol, 3.5 equiv.). The reaction was heated 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 crude material was purified by silica gel chromatography using 50% ethyl acetate / hexane to give (PA1.8) (17.0 g, 94.3%). MS (ES): m / z 330 [M+H] +

[0406] Step-4. 3-Bromocyclopent-2-en-1-one (PA1.1) A solution of cyclopentane-1,3-dione (PA1.0) (10 g, 10.19 mmol, 1.0 equiv) in DCM (240 mL) was treated portionwise with triphenylphosphine (29.1 g, 11.11 mmol, 1.0 equiv) and stirred at 0 °C for 30 min. Triethylamine (17 mL, 14.12 mmol, 2.0 equiv) was added slowly, followed by bromine (5.7 mL, 11.11 mmol, 1.1 equiv) at room temperature and stirred for 20 min. Upon completion, the DCM was removed under reduced pressure, and water (500 mL) was added slowly to the resulting residue, which was extracted with ethyl acetate (3 × 500 mL). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The crude residue was purified by column chromatography (2% gradient elution; ethyl acetate in hexane) to give PA1.1 (10 g, 96.88%), MS(ES): m / z 303 [M+1] + , was obtained.

[0407] Step-5. 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclopent-2-en-1-one (PA1.2) To a solution of 3-bromocyclopent-2-en-1-one (PA1.1) (1.5 g, 46.55 mmol, 1.0 equiv.) in dioxane (15 mL), potassium acetate (2.6 g, 39.65 mmol, 4 equiv.) was added, followed by the dropwise addition of bispinacoline diborane (2.98 g, 40.55 mmol, 3.0 equiv.) over 10 min while degassing with Ar. PdCl(dppf) (0.702 g, 32.74 mmol, 0.07 equiv.) was then added and stirred at 120 °C for 30 min. After completion, water (200 mL) was slowly added to the resulting residue, which was then extracted with ethyl acetate (3 × 200 mL). The combined organic layers were washed with brine, dried over NaSO, and concentrated under reduced pressure. The crude residue was purified by column chromatography (gradient elution with 10% ethyl acetate in hexane) to give PA1.2 (1.2 g, 96%), MS(ES): m / z 299 [M+1] + , was obtained.

[0408] Step-6. tert-Butyl (6-((dimethylamino)methyl)-5-(3-oxocyclopent-1-en-1-yl)pyridin-2-yl)carbamate (PA1.3) To a solution of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclopent-2-en-1-one (PA1.2) (1 g, 4.80 mmol, 1.0 equiv.) in dioxane (15 ml), potassium phosphate (4.07 g, 19.2 mmol, 3 equiv.) was added, followed by the dropwise addition of tert-butyl (5-bromo-6-((dimethylamino)methyl)pyridin-2-yl)carbamate (PA1.8) (3.17 g, 9.6 mmol, 2.0 equiv.). The reaction was degassed with Ar for 10 min. X-phose PdG2 (0.38 g, 0.4 mmol, 0.1 equiv.) was added and stirred at 80 °C for 1 h. After completion, water (50 ml) was slowly added to the resulting residue and extracted with ethyl acetate (3 x 25 ml). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The crude residue was purified by column chromatography (12-15% gradient elution with ethyl acetate in hexanes) to give PA1.3 (0.700 g, 43.95%), MS (ES): m / z 332.1 [M+1]. + , was obtained.

[0409] Step-7. tert-Butyl (6-((dimethylamino)methyl)-5-(3-oxocyclopentyl)pyridin-2-yl)carbamate (PA1.4) To a solution of (6-((dimethylamino)methyl)-5-(3-oxocyclopent-1-en-1-yl)pyridin-2-yl)carbamic acid (PA1.3) (0.580 g, 1.75 mmol, 1.0 equiv.) in THF (5.8 ml), acetic acid (0.4 ml) and ammonium formate (0.22 g, 3.5 mmol, 2.0 equiv.) were added, followed by palladium hydroxide (0.6 g) in an autoclave reaction under 20 mmbar pressure. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was filtered through a celite bed to give PA1.4 (0.58 g, 99.40%). MS (ES): m / z = 334.2 [M+H] +

[0410] Step-8. N-(6-(hydroxymethyl)-5-(3-methoxycyclopentyl)pyridin-2-yl)cyclopropanecarboxamide (PA1.5) A solution of tert-butyl (6-((dimethylamino)methyl)-5-(3-oxocyclopentyl)pyridin-2-yl)carbamate (PA1.4) (0.580 g, 1.73 mmol, 1.0 equiv) in ethanol (7 ml) was treated portionwise with sodium borohydride (0.397 g, 10.43 mmol, 6.0 equiv) and stirred at 70 °C for 30 minutes. Upon completion, the ethanol was removed under reduced pressure, water (50 ml) was added slowly to the resulting residue, and the reaction was extracted with ethyl acetate (3 × 25 ml). The combined organic layers were washed with brine, dried over NaSO, and concentrated under reduced pressure to give PA1.5 (0.55 g, 94.26%), MS (ES): m / z 336.5 [M+1]. + , was obtained.

[0411] Step-9. 3-(6-amino-2-((dimethylamino)methyl)pyridin-3-yl)cyclopentan-1-ol (PA1) A solution of N-(6-(hydroxymethyl)-5-(3-methoxycyclopentyl)pyridin-2-yl)cyclopropanecarboxamide (PA1.5) (0.550 g, 1.63 mmol, 1.0 equiv) in DCM (5 ml) was treated dropwise with 4 M HCl in dioxane (8.0 ml) and stirred at room temperature for 30 minutes. Upon completion, the DCM was removed under reduced pressure, and water (50 ml) was slowly added to the resulting residue, which was then extracted with ethyl acetate (2 x 25 ml). The organic layer was discarded, and the aqueous layer was basified with Na2CO3 and extracted with DCM (3 x 25 ml). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated under reduced pressure to give PA1 (0.2 g, 51.83%), MS (ES): m / z 236.5 [M +1]. + , was obtained.

[0412] Method PA2 - Preparation of 6-((dimethylamino)methyl)-5-(1,4-dioxan-2-yl)pyridin-2-amine (PA2) [ka] Step-1. 6-(cyclopropanecarboxamido)-3-(5,6-dihydro-1,4-dioxin-2-yl)methyl picolinate (PA2.1) A solution of methyl 3-bromo-6-(cyclopropanecarboxamido)picolinate (PA2.0) (2.0 g, 6.68 mmol, 1.0 equiv.) and tributyl(5,6-dihydro-1,4-dioxin-2-yl)stannane (2.5 g, 6.68 mmol, 1.0 equiv.) in 1,4-dioxane (20 mL) was degassed for 10-15 min. Bis(triphenylphosphine)palladium(II) dichloride (0.23 g, 0.33 mmol, 0.05 equiv.) and copper(I) iodide (0.120 g, 0.668 mmol, 0.1 equiv.) were added to the reaction mixture and stirred at 100 °C for 2 h. Upon completion, the reaction mixture was cooled to room temperature, poured into water (250 ml), extracted into ethyl acetate (3 x 150 ml), and the combined organic layers were washed with brine (200 ml), passed through Na2SO4, and concentrated under reduced pressure to give the crude material, which was further purified by column chromatography, eluting the compound with 40-50% ethyl acetate in hexane to give PA2.1 (0.870 g, 42.76%), MS (ES): m / z 305.1 [M+H]. + , was obtained.

[0413] Step-2. 6-(cyclopropanecarboxamido)-3-(1,4-dioxan-2-yl)methyl picolinate (PA2.2) To a solution of methyl 6-(cyclopropanecarboxamido)-3-(5,6-dihydro-1,4-dioxin-2-yl)picolinate (PA2.1) (0.870 g, 2.86 mmol, 1.0 equiv.) and 20% wet palladium hydroxide on carbon (0.7 g) in methanol (8 mL) and THF (8 mL) was added ammonium formate (0.36 g, 5.72 mmol, 2.0 equiv.) and acetic acid (0.7 mL) at room temperature. After 10 minutes, the reaction mixture was allowed to settle and stirred under hydrogen gas for 16 hours. The reaction mixture was then filtered through a pad of Celite, and the filtrate was concentrated under reduced pressure. The residue was basified with saturated NaHCO solution (30 mL) and extracted with DCM (3 × 25 mL). The combined organic layers were passed through NaSO and concentrated under reduced pressure to give PA2.2 (0.650 g, 74.22%). MS (ES): m / z 307.2 [M+H] +

[0414] Step-3. N-(5-(1,4-dioxan-2-yl)-6-(hydroxymethyl)pyridin-2-yl)cyclopropanecarboxamide (PA2.3) To a cooled solution of methyl 6-(cyclopropanecarboxamido)-3-(1,4-dioxan-2-yl)picolinate (PA2.2) (2.5 g, 8.16 mmol, 1.0 equiv.) in ethanol (25 ml) was added sodium borohydride (1.24 g, 32.64 mmol, 4.0 equiv.) in portions at 0 °C. The reaction mixture was stirred at room temperature for 1 h. The reaction mixture was concentrated under reduced pressure to give the crude product, which was diluted with water (70 ml) and extracted with dichloromethane (3 × 40 ml). The combined organic extracts were washed with brine (80 ml), then dried over Na SO , filtered, and concentrated under reduced pressure. The crude residue was purified by column chromatography using a gradient elution with 30% ethyl acetate in hexane to give PA2.3 (1.4 g, 61.64%). MS (ES): m / z 279.4 [M+H] +

[0415] Step-4, 5. N-(6-((dimethylamino)methyl)-5-(1,4-dioxan-2-yl)pyridin-2-yl)cyclopropanecarboxamide (PA2.4) To a solution of N-(5-(1,4-dioxan-2-yl)-6-(hydroxymethyl)pyridin-2-yl)cyclopropanecarboxamide (PA2.3) (0.650 g, 2.12 mmol, 1.0 equiv) in DCM (7 mL) was added N,N-diisopropylethylamine (1.3 mL, 7.42 mmol, 3.5 equiv) dropwise at 0 °C. The reaction was stirred for 20 min, followed by the dropwise addition of mesyl chloride (0.25 mL, 3.18 mmol, 1.5 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 20–25 min. Upon completion, the reaction mixture was quenched with DM water (60 mL) and extracted with DCM (3 × 30 mL). The combined organic layers were washed with brine (30 ml), passed through Na2SO4 and concentrated under reduced pressure to give the mesylated product (0.8 g crude, 89.80%). MS (ES): m / z 357.3 [M+1] + To a solution of the above mesylated product (0.8 g, 2.24 mmol, 1.0 equiv.) in acetonitrile (8 ml), N,N-diisopropylethylamine (1.6 ml, 8.96 mmol, 4.0 equiv.) was added dropwise, followed by the addition of dimethylamine hydrochloride (0.38 g, 4.48 mmol, 2.0 equiv.) at room temperature. The reaction mixture was stirred at 90 °C for 1 h. Upon completion, the reaction mixture was quenched with DM water (80 ml) and extracted with DCM (3 × 30 ml). The combined organic layers were washed with brine (50 ml), dried over NaSO, and concentrated under reduced pressure to give the crude material. This crude material was further purified by column chromatography, and the compound was eluted with 50–60% ethyl acetate in hexane to give PA2.4 (0.450 g, 77.25%). MS (ES): m / z 306.2 [M+H] +

[0416] Step-6. 6-((dimethylamino)methyl)-5-(1,4-dioxan-2-yl)pyridin-2-amine (PA2) To a solution of N-(6-((dimethylamino)methyl)-5-(1,4-dioxan-2-yl)pyridin-2-yl)cyclopropanecarboxamide (PA2.4) (0.450 g, 1.47 mmol, 1.0 equiv.) in methanol (3.6 ml) and water (1 ml), sodium hydroxide (0.588 g, 14.7 mmol, 10.0 equiv.) was added. The reaction mixture was stirred at 80 °C for 16 h. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure to remove methanol. The resulting residue was diluted with water (60 ml) and extracted with ethyl acetate (3 × 40 ml). The combined organic extracts were washed with brine (50 ml), dried over NaSO, filtered, and concentrated under reduced pressure to give PA2 (0.230 g, 65.77%). The crude material was used in the next step without purification. MS (ES): m / z 238.2 [M+H] + .

[0417] Method PA3 - Preparation of 6-((dimethylamino)methyl)-5-(2-methyltetrahydrofuran-2-yl)pyridin-2-amine (PA3) [ka] Step 1. tert-Butylbenzyl (6-((benzyloxy)methyl)-5-(2-methyltetrahydrofuran-2-yl)pyridin-2-yl)carbamate (3.1) To a cooled solution of tert-butylbenzyl (6-((benzyloxy)methyl)-5-bromopyridin-2-yl)carbamate (PA3.0) (20 g, 41.40 mmol, 1.0 equiv.) in dry THF (200 ml), 5-chloropentan-2-one (7.4 g, 62.1 mmol, 1.5 equiv.) was added and the reaction mixture was stirred at −78° C. for 10 minutes. n-Butyllithium (2.5 M in hexanes) (41.5 ml, 103.5 mmol, 2.5 equiv.) was then added and stirred at room temperature for 2 hours. Upon completion, the reaction mixture was quenched with water (600 ml) and the product was extracted with ethyl acetate (3×200 ml). The combined organic layers were washed with brine solution (300 ml), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. The crude was further purified by column chromatography, and the compound was eluted with 5-10% ethyl acetate in hexane to give PA3.1 (5.0 g, 24.73%). MS (ES): m / z 489.2 [M+H] +

[0418] Step-2. (6-(benzylamino)-3-(2-methyltetrahydrofuran-2-yl)pyridin-2-yl)methanol (PA3.2) To a cooled solution of tert-butylbenzyl (6-((benzyloxy)methyl)-5-(2-methyltetrahydrofuran-2-yl)pyridin-2-yl)carbamate (PA3.1) (5.0 g, 10.24 mmol, 1.0 equiv.) in DCM (50 ml) at 0 °C was added Trifflic acid (10 ml) dropwise. The reaction mixture was stirred at 0 °C for 15 min. Upon completion, the reaction mixture was quenched in saturated NaHCO solution (60 ml) and extracted with dichloromethane (3 × 35 ml). The organic layers were combined, dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was purified by column chromatography, and the compound was eluted with 50-55% ethyl acetate in hexane to give PA3.2 (1.7 g, 55.68%). MS (ES): m / z 299.1 [M+H] +

[0419] Step-3, 4. N-Benzyl-6-((dimethylamino)methyl)-5-(2-methyltetrahydrofuran-2-yl)pyridin-2-amine (PA3.3) To a solution of (6-(benzylamino)-3-(2-methyltetrahydrofuran-2-yl)pyridin-2-yl)methanol (PA3.2) (0.650 g, 2.12 mmol, 1.0 equiv.) in DCM (7 ml), N,N-diisopropylethylamine (1.3 ml, 7.42 mmol, 3.5 equiv.) was added dropwise at 0 °C and stirred for 20 min, followed by the dropwise addition of mesyl chloride (0.25 ml, 3.18 mmol, 1.5 equiv.) at 0 °C. The reaction mixture was stirred at 0 °C for 20–25 min. After completion, the reaction mixture was quenched with DM water (60 ml) and extracted with DCM (3 × 30 ml). The combined organic layers were washed with brine (30 ml), passed through Na2SO4 and concentrated under reduced pressure to give the mesylated product (0.8 g crude, 89.80%), MS (ES): m / z 357.3 [M+1]. + , afforded. To a solution of the above mesylated product (0.8 g, 2.24 mmol, 1.0 equiv.) in acetonitrile (8 ml), N,N-diisopropylethylamine (1.6 ml, 8.96 mmol, 4.0 equiv.) was added dropwise, followed by the addition of dimethylamine hydrochloride (0.38 g, 4.48 mmol, 2.0 equiv.) at room temperature. The reaction mixture was stirred at 90 °C for 1 h. Upon completion, the reaction mixture was quenched with DM water (80 ml) and extracted with DCM (3 × 30 ml). The combined organic layers were washed with brine (50 ml), dried over Na2SO4, and concentrated under reduced pressure to afford the crude material. This was further purified by column chromatography, and the compound was eluted with 50–60% ethyl acetate in hexane to afford PA3.3 (0.450 g, 77.25%). MS (ES): m / z 306.2 [M+H] +

[0420] Step-5. 6-((dimethylamino)methyl)-5-(2-methyltetrahydrofuran-2-yl)pyridin-2-amine (PA3) To a solution of N-benzyl-6-((dimethylamino)methyl)-5-(2-methyltetrahydrofuran-2-yl)pyridin-2-amine (PA3.3) (1.8 g, 5.53 mmol, 1.0 equiv) in DCM (20 ml) was added Trifflic acid (3 ml) dropwise at 0 °C. The reaction mixture was stirred at 0 °C for 15 minutes. Upon completion, the reaction mixture was quenched with saturated NaHCO solution (25 ml) and the product was extracted with 10% methanol in dichloromethane (3 × 20 ml). The organic layers were combined, washed with brine solution (25 ml), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by trituration with hexane and diethyl ether to give PA3 (0.7 g, 53.78%). MS (ES): m / z 236.1 [M+H] +

[0421] Method PA4 - Preparation of 6-((S)-1-methylpyrrolidin-2-yl)-5-(tetrahydrofuran-3-yl)pyridin-2-amine (PA4) [ka] Step 1. 3-Bromo-6-(cyclopropanecarboxamide)picolinic acid (4.1) To a solution of methyl 3-bromo-6-(cyclopropanecarboxamide)picolinate (PA4.0) (27 g, 90.60 mmol, 1.0 equiv.) in methanol:THF:water (120 ml:120 ml:25 ml) was added sodium hydroxide (10.8 g, 271.8 mmol, 3.0 equiv.). The reaction was stirred at room temperature for 6 hours. Upon completion, the reaction mixture was concentrated under reduced pressure and acidified with citric acid solution to give a precipitate, which was filtered and concentrated under high vacuum to give PA4.1, which was used in the next step without purification. (24 g, 93.26%), MS (ES): m / z 285.1 [M+H] +

[0422] Step-2. 3-Bromo-6-(cyclopropanecarboxamido)-N-methoxy-N-methylpicolinamide (PA4.2) To a solution of 3-bromo-6-(cyclopropanecarboxamido)picolinic acid (PA4.1) (24 g, 84.21 mmol, 1.0 equiv.) and N,O-dimethylhydroxylamine hydrochloride (9.8 g, 101.052 mmol, 1.2 equiv.) in N,N-dimethylformamide (200 mL), hydroxybenzotriazole (17.0 g, 126.31 mmol, 1.5 equiv.), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (32 g, 210.52 mmol, 2.5 equiv.), and trimethylamine (35.5 mL, 252.63 mmol, 3.0 equiv.) were added. The reaction was stirred at room temperature for 16 hours. Upon completion, the reaction mixture was transferred to water (850 mL) and the product was extracted with ethyl acetate (3 x 250 mL). The organic layers were combined, washed with brine solution (500 ml), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material, which was further purified by column chromatography; the compound was eluted with 20% ethyl acetate in hexane to give PA4.2 (16 g, 57.92%). MS (ES): m / z 329.02 [M+H] +

[0423] Step-3. N-(6-(3-(1,3-dioxan-2-yl)propanoyl)-5-bromopyridin-2-yl)cyclopropanecarboxamide (PA4.4) To a cooled solution of 3-bromo-6-(cyclopropanecarboxamido)-N-methoxy-N-methylpicolinamide (PA4.2) (4.0 g, 12.19 mmol, 1.0 equiv.) in dry THF (40 ml), (2-(1,3-dioxan-2-yl)ethyl)magnesium bromide (PA4.3) (0.5 M in THF) (48 ml, 24.38 mmol, 2.0 equiv.) was added dropwise at 0° C. under a N atmosphere. The reaction was stirred at room temperature for 8 hours. Upon completion, the reaction mixture was poured into water (200 ml) and the product was extracted with ethyl acetate (3×80 ml). The organic layers were combined, washed with brine solution (120 ml), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, and the compound was eluted with 25% ethyl acetate in hexane to give PA4.4 (2.3 g, 49.24%). MS (ES): m / z 384.05 [M+H] +

[0424] Step-4. N-(5-bromo-6-(4-oxobutanoyl)pyridin-2-yl)cyclopropanecarboxamide (4.5) To a solution of N-(6-(3-(1,3-dioxan-2-yl)propanoyl)-5-bromopyridin-2-yl)cyclopropanecarboxamide (PA4.4) (2.3 g, 6.00 mmol, 1.0 equiv) in THF (30 mL) was added formic acid (30 mL). The reaction was stirred at 50-60 °C for 48 h. Upon completion, the reaction mixture was concentrated under reduced pressure, poured into saturated NaHCO solution (50 mL), and extracted with dichloromethane (3 x 35 mL). The organic layers were combined, dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by trituration with hexanes followed by diethyl ether to give pure PA4.5 (1.9 g, 97.36%). MS (ES): m / z 326.01 [M+H] +

[0425] Step-5. N-(5-bromo-6-((S)-1-((R)-1-(4-methoxyphenyl)ethyl)pyrrolidin-2-yl)pyridin-2-yl)cyclopropanecarboxamide (PA4.7) A solution of N-(5-bromo-6-(4-oxobutanoyl)pyridin-2-yl)cyclopropanecarboxamide (PA4.5) (1.6 g, 4.92 mol, 1.0 equiv.) in dry THF (15 ml) was cooled to −70° C., and sodium triacetoxyborohydride (3.1 g, 14.76 mmol, 3.0 equiv.) and acetic acid (0.8 ml) were added under a N atmosphere. The reaction was stirred at −70° C. for 30 minutes, and then (R)-1-(4-methoxyphenyl)ethan-1-amine (PA4.6) (0.816 g, 5.41 mmol, 1.1 equiv.) dissolved in THF (2 ml) was added to the reaction mixture and stirred at room temperature for 16 hours. Upon completion, the reaction mixture was poured into saturated NaHCO solution (50 ml), and the product was extracted with ethyl acetate (3×25 ml). The organic layers were combined, washed with brine solution (30 ml), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material, which was further purified by column chromatography; the compound was eluted with 80% ethyl acetate in hexane to give PA4.7 (0.8 g, 36.59%). MS (ES): m / z 445.1 [M+H] +

[0426] Step-6. (S)—N-(5-bromo-6-(pyrrolidin-2-yl)pyridin-2-yl)cyclopropanecarboxamide (PA4.8) A solution of N-(5-bromo-6-((S)-1-((R)-1-(4-methoxyphenyl)ethyl)pyrrolidin-2-yl)pyridin-2-yl)cyclopropanecarboxamide (PA4.7) (0.8 g, 1.80 mmol, 1.0 equiv) in TFA (8 ml) was stirred at 50 °C for 16 hours. After completion, the reaction mixture was basified with saturated NaHCO solution (30 ml) and the product was extracted with ethyl acetate (3 x 20 ml). The organic layers were combined, washed with brine solution (25 ml), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography on neutral alumina, and the compound was eluted with 56% ethyl acetate in hexane to give PA4.8 (0.4 g, 71.63%). MS (ES): m / z 311.05 [M+H] +

[0427] Step-7. (S)—N-(5-bromo-6-(1-methylpyrrolidin-2-yl)pyridin-2-yl)cyclopropanecarboxamide (PA4.9) To a cooled solution of (S)—N-(5-bromo-6-(pyrrolidin-2-yl)pyridin-2-yl)cyclopropanecarboxamide (PA4.8) (0.3 g, 0.96 mmol, 1.0 equiv) in 1,2-dichloroethane (3 ml) was added paraformaldehyde (0.115 g, 3.84 mmol, 4.0 equiv) at 0° C. Acetic acid (3.5 ml) was added at 0° C. and stirred at room temperature for 1 hour, followed by the addition of sodium triacetoxyborohydride (1.0 g, 4.8 mmol, 5.0 equiv) in portions at 0° C. and the reaction was stirred at room temperature for 16 hours. Upon completion, the reaction mixture was poured into water (50 ml) and the product was extracted with DCM (3×50 mL). The combined organic layers were washed with brine solution (20 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, and the compound was eluted with 20% ethyl acetate in hexane to give PA4.9 (0.310 g, 31.89%). MS (ES): m / z 324.2 [M+H] +

[0428] Step-8. (S)—N-(5-furan-3-yl)-6-(1-methylpyrrolidin-2-yl)pyridin-2-yl)cyclopropanecarboxamide (4.11) (S)—N-(5-bromo-6-(1-methylpyrrolidin-2-yl)pyridin-2-yl)cyclopropanecarboxamide (PA4.9) (0.310 g, 0.95 mmol, 1.0 equiv.), furan-3-ylboronic acid (PA4.10) (0.212 g, 1.9 mmol, 2 equiv.) and potassium phosphate, tribasic (0.6 g) in 1-4 dioxane:water (6 mL:2 mL). A solution of PA4.11 (2.85 mmol, 3.0 equiv.) was degassed with nitrogen for 20 minutes, and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (CAS No. 1310584-14-5) (0.074 g, 0.095 mmol, 0.1 equiv.) was added at room temperature. The reaction mixture was heated at 100°C for 15 minutes. Upon completion, the reaction was cooled to room temperature. The aqueous layer was separated from the reaction mixture, and the organic layer was filtered through a bed of Celite and concentrated under reduced pressure. The crude material was purified by silica gel chromatography, and the product was eluted with 3.0% methanol in dichloromethane to give PA4.11 (0.210 g, 70.53%). MS (ES): m / z 312.1 [M+H] +

[0429] Step-9. N-(6-((S)-1-methylpyrrolidin-2-yl)-5-(tetrahydrofuran-3-yl)pyridin-2-yl)cyclopropanecarboxamide (4.12) A solution of (S)—N-(5-(furan-3-yl)-6-(1-methylpyrrolidin-2-yl)pyridin-2-yl)cyclopropanecarboxamide (PA4.11) (0.240 g, 0.77 mmol, 1.0 equiv) in methanol (3 mL) and THF (1 mL) was treated with ammonium formate (0.194 g, 3.08 mmol, 4.0 equiv), acetic acid (0.1 mL, 0.7 V), and 20% wet palladium hydroxide on carbon (0.168 g, 1:0.7 w / w) at room temperature. The reaction mixture was stirred under a hydrogen gas atmosphere at room temperature for 16 hours. The progress of the reaction was monitored by LCMS. The reaction mixture was filtered through a bed of Celite, and the filtrate was concentrated under reduced pressure. The residue was basified with saturated NaHCO3 (25 mL) solution and extracted with DCM (3 x 15 mL), and the combined organic layers were dried, filtered, and concentrated to give PA4.12 (0.180 g, 74.07%), which was used in the next step without purification. MS (ES): m / z 316.2 [M+H] +

[0430] Step-10. 6-((S)-1-methylpyrrolidin-2-yl)-5-(tetrahydrofuran-3-yl)pyridin-2-amine (PA4) To a solution of N-(6-((S)-1-methylpyrrolidin-2-yl)-5-(tetrahydrofuran-3-yl)pyridin-2-yl)cyclopropanecarboxamide (PA4.12) (0.180 g, 0.56 mmol, 1.0 equiv) in methanol:water (2 mL:2 mL) was added sodium hydroxide (0.224 g, 5.6 mmol, 3.0 equiv). The reaction was stirred at 60° C. for 16 h. Upon completion, the reaction mixture was poured into DM water (50 mL) and the product was extracted with 10% methanol in DCM (3×40 mL). The organic layers were combined, dried over sodium sulfate, and concentrated under reduced pressure to give the crude material, which was purified by trituration with n-hexane (5 ml) followed by diethyl ether (5 ml) to give PA4 (0.1 g, 70.85%), MS (ES): m / z 248.1 [M+H]. + , was obtained.

[0431] Method PA5 Preparation of (7-chloro-4-methyl-2,3,4,5-tetrahydro-1H-pyrido[3,2-e][1,4]diazepin-1-yl)(tetrahydrofuran-3-yl)methanone (PA5) [ka] Step 1. tert-Butyl (2-(((6-chloro-3-fluoropyridin-2-yl)methyl)(methyl)amino)ethyl)carbamate (5.1) To a solution of 6-chloro-3-fluoropicolinaldehyde (PA5.0) (10 g, 62.89 mmol, 1.0 equiv.) and tert-butyl (2-(methylamino)ethyl)carbamate (16.4 g, 94.33 mmol, 1.5 equiv.) in 1,2-dichloroethane (100 mL), acetic acid (2 mL) was added and cooled to 0 °C. The reaction mixture was stirred at 0 °C for 15-20 min. Sodium triacetoxyborohydride (39.9 g, 188.67 mmol, 3.0 equiv.) was then added portionwise at 0 °C and stirred at the same temperature for 1 h. Upon completion, the reaction mixture was quenched with ice-cold water (200 ml) and extracted with ethyl acetate (3 × 120 mL). The combined organic layers were washed with brine (150 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, eluting the compound with 16% ethyl acetate in hexane to give PA5.1 (12.0 g, 60.25%) as an oily white solid. MS (ES): m / z 318.2 [M+H] +

[0432] Step—2,3.7-Chloro-4-methyl-2,3,4,5-tetrahydro-1H-pyrido[3,2-e][1,4]diazepine (PA5.2) To a cooled solution of tert-butyl (2-(((6-chloro-3-fluoropyridin-2-yl)methyl)(methyl)amino)ethyl)carbamate PA5.1 (12.0 g, 37.85 mmol, 1.0 equiv.) in DCM (120 mL), trifluoroacetic acid (58 mL, 757.0 mmol, 20.0 equiv.) was added at 0 °C and stirred for 30 min. The reaction mixture was concentrated under reduced pressure at 35-40 °C to give the TFA salt. The TFA salt (8.0 g, 36.86 mmol, 1.0 equiv.) was dissolved in dimethyl sulfoxide (50 mL), and N,N-diisopropylethylamine (45 mL, 258.02 mmol, 7.0 equiv.) was added and stirred at 110 °C for 16 h. After completion, the reaction mixture was cooled to room temperature, poured into DM water (350 mL), and extracted with 10% methanol in dichloromethane (3 x 150 mL). The combined organic layers were washed with brine solution (150 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, and the compound was eluted with 2.5% methanol in dichloromethane to give PA5.2 (11.52 g, 0.860%). MS (ES): m / z 198.08 [M+H] +

[0433] Step-4 (7-chloro-4-methyl-2,3,4,5-tetrahydro-1H-pyrido[3,2-e][1,4]diazepin-1-yl)(tetrahydrofuran-3-yl)methanone (PA5) To a cooled solution of tetrahydrofuran-3-carboxylic acid (0.860 g, 7.41 mmol, 1.0 equiv.) in DCM (9 mL) was added dropwise oxalyl chloride (1.2 mL, 14.82 mmol, 2.0 equiv.) and catalytic DMF (1 mL) at 0 °C. The reaction was stirred for 30 min and then concentrated under reduced pressure at 35–40 °C to give the crude material. The crude material (0.5 g, 3.70 mmol, 1.0 equiv.) was dissolved in DCM (5 mL). 7-Chloro-4-methyl-2,3,4,5-tetrahydro-1H-pyrido[3,2-e][1,4]diazepine (PA5.2) (0.364 g, 1.85 mmol, 0.5 equiv.) and triethylamine (0.6 mL, 4.44 mmol, 1.2 equiv.) were added to the reaction mixture and stirred at room temperature for 5 h. After completion, the reaction mixture was poured into DM water (60 mL) and the product was extracted with 10% methanol in dichloromethane (3 x 30 mL). The combined organic layers were washed with brine solution (50 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, and the compound was eluted with 2.1% methanol in dichloromethane to give PA5 (0.710 g, 55.17%). MS (ES): m / z 296.1 [M+H] +

[0434] Method PA6 - Preparation of 1-(6-bromo-3-((1r,3r)-3-methoxycyclobutoxy)pyridin-2-yl)-N,N-dimethylmethanamine (PA6) [ka] Step 1. Methyl 6-bromo-3-hydroxypicolinate (PA6.1) To a solution of methyl 3-hydroxypicolinate (PA6.0) (15 g, 98.03 mmol, 1.0 equiv.) in DM water (100 mL) was added bromine (6.0 mL, 117.63 mmol, 1.2 equiv.) dropwise at 0 °C and stirred at 0 °C for 2 h. Upon completion, the reaction mixture was quenched in sodium thiosulfate solution (300 mL) and the product was extracted with DCM (4 × 160 mL). The organic layers were combined, washed with brine solution (200 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography; the compound was eluted with 10–15% ethyl acetate in hexane to give PA6.1 (8.0 g, 35.20%). MS (ES): m / z 232.9 [M+H] +

[0435] Step 2. Methyl 3-(benzyloxy)-6-bromopicolinate (6.2) To a cooled solution of methyl 6-bromo-3-hydroxypicolinate (PA6.1) (8.0 g, 34.48 mmol, 1.0 equiv.) in acetonitrile (80 mL) was added potassium carbonate (14.2 g, 103.44 mmol, 3.0 equiv.). Benzyl bromide (8.1 mL, 68.96 mmol, 2.0 equiv.) was added dropwise at 0 °C and stirred at 80 °C for 2 h. Upon completion, the reaction mixture was quenched with sodium bicarbonate solution (200 mL), and the product was extracted with DCM (3 × 120 mL). The organic layers were combined, washed with brine solution (200 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography; the compound was eluted with 5–10% ethyl acetate in hexane to give PA6.2 (7.5 g, 67.52%). MS(ES): m / z 323.00[M+H] +

[0436] Step-3. (3-(benzyloxy)-6-bromopyridin-2-yl)methanol (PA6.3) To a solution of methyl 3-(benzyloxy)-6-bromopicolinate (PA6.2) (7.5 g, 23.29 mmol, 1.0 equiv.) in ethanol (75 mL) was added sodium borohydride (61.5 g, 69.87 mmol, 3.0 equiv.) in small portions at 0 °C under a N atmosphere and stirred at room temperature for 1 h at 70 °C. Upon completion, the reaction mixture was concentrated under reduced pressure to remove ethanol, then diluted with water (100 mL), and the product was extracted with ethyl acetate (3 × 60 mL). The organic layers were combined, washed with brine solution (80 mL), dried over sodium sulfate, and concentrated under reduced pressure to give PA6.3 (5.2 g, 75.94%). MS (ES): m / z 295.00 [M+H] +

[0437] Step-4, 5. 1-(3-(benzyloxy)-6-bromopyridin-2-yl)-N,N-dimethylmethanamine (PA6.4) To a solution of (3-(benzyloxy)-6-bromopyridin-2-yl)methanol (PA6.3) (6.6 g, 22.44 mmol, 1.0 equiv) in DCM (70 mL) was added N,N-diisopropylethylamine (13.5 mL, 78.57 mmol, 3.5 equiv) dropwise at 0 °C and stirred for 20 min, followed by the dropwise addition of mesyl chloride (2.6 mL, 33.66 mmol, 1.5 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 20-25 min. Upon completion, the reaction mixture was quenched with DM water (80 mL) and extracted with DCM (3 × 30 mL). The combined organic layers were washed with brine (50 mL), passed through NaSO, and concentrated under reduced pressure to afford the mesylated product (7.5 g crude, 89.80%). MS(ES): m / z 372.1[M+1] +To a solution of the above mesylated product (7.5 g, 20.16 mmol, 1.0 equiv.) in acetonitrile (75 mL), N,N-diisopropylethylamine (10.5 mL, 80.64 mmol, 4.0 equiv.) was added dropwise, followed by the dropwise addition of dimethylamine hydrochloride (3.2 g, 40.32 mmol, 2.0 equiv.) at room temperature. The reaction mixture was stirred at 90 °C for 1 h. Upon completion, the reaction mixture was quenched in DM water (180 mL) and extracted with DCM (3 × 50 mL). The combined organic layers were washed with brine (120 mL), dried over NaSO, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography; the compound was eluted with 50–60% ethyl acetate in hexane to give PA6.4 (5.0 g, 77.25%). MS (ES): m / z 322.05 [M+H] +

[0438] Step-6. 6-Bromo-2-((dimethylamino)methyl)pyridin-3-ol (PA6.5) To a solution of 1-(3-(benzyloxy)-6-bromopyridin-2-yl)-N,N-dimethylmethanamine (PA6.4) (1.0 g, 3.11 mmol, 1.0 equiv) in DCM (10 mL) was added triflic acid (2.0 mL) at 0° C. and stirred at room temperature for 30 minutes. After completion, the reaction mixture was diluted with water (100 mL) and extracted with DCM (2×40 mL). The aqueous layer was quenched with aqueous sodium bicarbonate solution, and the product was extracted with 10% methanol in DCM (3×50 mL). The organic layers were combined, washed with brine solution (80 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, and the compound was eluted with 70% ethyl acetate in hexane to give PA6.5 (0.450 g, 62.55%). MS(ES): m / z 232.2[M+H] +

[0439] Step 7. (1s,3s)-3-Methoxycyclobutyl 4-methylbenzenesulfonate (PA6.7) To a solution of (1s,3s)-3-methoxycyclobutan-1-ol (PA6.6) (0.3 g, 2.94 mmol, 1.0 equiv) in DCM (6 mL) was added triethylamine (0.4 mL, 2.94 mmol, 1.0 equiv), followed by 4-dimethylaminopyridine (0.035 g, 0.29 mmol, 0.1 equiv). The reaction mixture was cooled to 0 °C, 4-toluenesulfonyl chloride (0.55 g, 2.94 mmol, 1.0 equiv) was added, and the reaction mixture was stirred at 50 °C for 16 h. Upon completion, the reaction mixture was quenched with sodium bicarbonate solution (50 mL) and extracted with ethyl acetate (2 × 25 mL). The organic layers were combined, washed with brine solution (40 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, and the compound was eluted with 15-20% ethyl acetate in hexane to give PA6.7 (0.450 g, 59.77%). MS (ES): m / z 257.08 [M+H] +

[0440] Step-8. 1-(6-bromo-3-((1r,3r)-3-methoxycyclobutoxy)pyridin-2-yl)-N,N-dimethylmethanamine (PA6) To a solution of 6-bromo-2-((dimethylamino)methyl)pyridin-3-ol (PA6.5) (0.250 g, 1.08 mmol, 1.0 equiv) and (1s,3s)-3-methoxycyclobutyl 4-methylbenzenesulfonate (PA6.7) (0.413 g, 1.62 mmol, 1.5 equiv) in dimethylformamide (3 mL) was added potassium carbonate (0.447 g, 3.24 mmol, 3.0 equiv). The reaction mixture was stirred at 90 °C for 16 h. Upon completion, the reaction mixture was transferred to ice-cold water (70 mL) and extracted with ethyl acetate (2 × 30 mL). The organic layers were combined, washed with brine solution (50 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, and the compound was eluted with 4.5-5% methanol in dichloromethane to give PA6 (0.4 g, 99.71%). MS (ES): m / z 316.2 [M+H] +

[0441] Method PA7. Preparation of 6-chloro-3-(3-methoxytetrahydrofuran-3-yl)picolinonitrile (PA7) [ka] Step-1. 2-Chloro-5-(3-methoxytetrahydrofuran-3-yl)pyridine (PA7.1) To a solution of 3-(6-chloropyridin-3-yl)tetrahydrofuran-3-ol (PA7.0) (0.5 g, 2.51 mmol, 1.0 equiv.) in dimethylformamide (5 mL) was added sodium hydride (60% in mineral oil) (0.180 g, 7.53 mmol, 3.0 equiv.) in small portions at 0 °C. Methyl iodide (0.3 mL, 5.02 mmol, 2.0 equiv.) was added dropwise and stirred at room temperature for 20 minutes. Upon completion, the reaction mixture was transferred to ice-cold water (80 mL) and the product was extracted with ethyl acetate (3 × 30 mL). The organic layers were combined, washed with brine solution (50 mL), dried over sodium sulfate, and concentrated under reduced pressure to give PA7.1 (0.450 g, 84.09%), MS (ES): m / z 214.06 [M+H]. + , was obtained.

[0442] Step-2. 2-Chloro-5-(3-methoxytetrahydrofuran-3-yl)pyridine 1-oxide (PA7.2) To a solution of 2-chloro-5-(3-methoxytetrahydrofuran-3-yl)pyridine (PA7.1) (0.450 g, 2.11 mmol, 2.0 equiv.) in DCM (8 mL), hydrogen peroxide-urea (CAS no. 124-43-6) (1.1 g, 12.66 mmol, 6.0 equiv.) was added slowly in small portions, and trifluoroacetic anhydride (1.8 mL, 12.66 mmol, 6.0 equiv.) was added at 0 °C. The reaction was stirred at room temperature for 1 h. Upon completion, the reaction mixture was transferred to a sodium sulfite solution (30 mL), and the product was extracted with dichloromethane (3 × 25 mL). The organic layers were combined, dried over sodium sulfate, and concentrated under reduced pressure to give the crude material, which was used in the next step without purification. PA7.2 (0.340 g crude, quant. %). MS (ES): m / z 230.05 [M+H] +

[0443] Step-3. 6-Chloro-3-(3-methoxytetrahydrofuran-3-yl)picolinonitrile (PA7) To a solution of 2-chloro-5-(3-methoxytetrahydrofuran-3-yl)pyridine 1-oxide (PA7.2) (0.340 g, 1.48 mmol, 2.0 equiv.) in acetonitrile (5 mL), triethylamine (0.6 mL, 4.45 mmol, 3.0 equiv.) and trimethylsilyl cyanide (0.43 g, 4.44 mmol, 3.0 equiv.) were added at room temperature. The reaction was stirred at 110 °C for 16 h. Upon completion, the reaction mixture was transferred to water (50 mL), and the product was extracted with ethyl acetate (4 × 20 mL). The organic layers were combined, washed with brine solution (30 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography; the compound was eluted with 8–10% ethyl acetate in hexane to give PA7 (0.240 g, 67.92%). MS(ES): m / z 239.2[M+H] +

[0444] Method PA8 - Preparation of 5-cyclopentyl-6-((dimethylamino)methyl)pyridin-2-amine (PA8) [ka] Step 1. tert-Butyl (5-(cyclopent-1-en-1-yl)-6-((dimethylamino)methyl)pyridin-2-yl)carbamate (PA8.1) A solution of tert-butyl (5-bromo-6-((dimethylamino)methyl)pyridin-2-yl)carbamate (PA8.0) (4.0 g, 12.12 mmol, 1.0 equiv.), cyclopent-1-en-1-ylboronic acid (2.7 g, 24.24 mmol, 2.0 equiv.), and potassium phosphate tribasic (7.7 g, 36.36 mmol, 3.0 equiv.) suspended in 1,4-dioxane (32 mL) and water (8 mL) was degassed with nitrogen for 20 minutes. The mixture was treated with chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (CAS number 1310584-14-5) (0.952 g, 1.21 mmol, 0.1 equiv.) and stirred at 120 °C for 1 h. Upon completion, the reaction mixture was transferred to DM water (100 mL) and the product was extracted with ethyl acetate (3 × 40 mL). The organic layers were combined, washed with brine solution (80 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography; the compound was eluted with 5–6% methanol in dichloromethane to give PA8.1 (3.5 g, 91.03%) as an oily white solid. MS (ES): m / z 318.2 [M+H] +

[0445] Step 2. tert-Butyl (5-(cyclopentyl-6-((dimethylamino)methyl)pyridin-2-yl)carbamate (PA8.2) To a solution of tert-butyl (5-(cyclopent-1-en-1-yl)-6-((dimethylamino)methyl)pyridin-2-yl)carbamate (PA8.1) (3.5 g, 11.04 mmol, 1.0 equiv.) and 20% wet palladium hydroxide on carbon (3.5 g) in methanol (25 mL) and THF (10 mL) was added ammonium formate (2.0 g, 33.12 mmol, 3.0 equiv.) and acetic acid (1.7 mL) at room temperature. After 10 minutes, the mixture was allowed to settle and stirred under a hydrogen gas atmosphere for 8 hours. The reaction mixture was filtered through a pad of Celite, and the filtrate was concentrated under reduced pressure. The resulting residue was basified with saturated NaHCO3 (80 mL) solution and extracted with DCM (3 × 40 mL) to give pure PA8.2 (3.0 g, 85.17%). MS(ES): m / z 320.2 [M+H] +

[0446] Step-3. 5-Cyclopentyl-6-((dimethylamino)methyl)pyridin-2-amine (PA8) To a solution of tert-butyl (5-cyclopentyl-6-((dimethylamino)methyl)pyridin-2-yl)carbamate (PA8.2) (3.0 g, 9.40 mmol, 1.0 equiv) in DCM (30 mL) was added trifluoroacetic acid (12 mL) at room temperature and stirred at 50 °C for 2 h. Upon completion, the reaction mixture was concentrated under reduced pressure to give the crude material, which was diluted with water (50 mL) and extracted with dichloromethane (3 × 30 mL). The aqueous layer was basified with sodium hydroxide, and the product was extracted with 10% methanol in dichloromethane (3 × 30 mL). The organic layers were combined, washed with brine solution (40 mL), dried over sodium sulfate, and concentrated under reduced pressure to give PA8 (1.2 g, 58.26%). MS (ES): m / z 220.2 [M+H] +

[0447] Method PA9 - Preparation of 6-chloro-3-(2,2-dimethyl-3-oxomorpholino)picolinonitrile (PA9) [ka] Step-1. 2-(benzylamino)ethan-1-ol (PA9.1) To a solution of 2-aminoethan-1-ol (20.0 g, 327.86 mmol, 1.0 equiv) in methanol (200 mL) was added benzaldehyde (PA9.0) (34.7 g, 327.86 mmol, 1.0 equiv) and sodium borohydride (6.2 g, 163.93 mmol, 0.5 equiv) at 0 °C. The reaction was stirred at room temperature for 6 h. Upon completion, the reaction mixture was concentrated under reduced pressure to remove methanol and then diluted with DM water (500 mL), and the product was extracted with ethyl acetate (3 × 200 mL). The organic layers were combined, washed with brine solution (400 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, and the compound was eluted with 5-6% methanol in dichloromethane to give PA9.1 (16.2 g, 32.72%), MS(ES): m / z 152.2 [M+H] + , was obtained.

[0448] Step-2. N-benzyl-2-bromo-N-(2-hydroxyethyl)-2-methylpropanamide (PA9.2) To a solution of 2-(benzylamino)ethan-1-ol (PA9.1) (16.2 g, 107.28 mmol, 1.0 equiv) in DCM (35 mL) was added trimethylamine (22.5 mL, 160.92 mmol, 1.5 equiv) at 0 °C, and the reaction was stirred at 0 °C for 15 min. 2-Bromo-2-methylpropanoyl bromide (24.5 g, 107.28 mmol, 1.0 equiv) was added and stirred at 0 °C for 2 h. Upon completion, the reaction mixture was transferred to DM water (400 mL), and the product was extracted with DCM (3 × 150 mL). The organic layers were combined, washed with brine solution (300 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, and the compound was eluted with 25% ethyl acetate in hexane to give PA9.2 (4.9 g, 15.24%), MS(ES): m / z 301.05 [M+H] + , was obtained.

[0449] Step-3. 4-Benzyl-2,2-dimethylmorpholin-3-one (PA9.3) To a solution of N-benzyl-2-bromo-N-(2-hydroxyethyl)-2-methylpropanamide (PA9.2) (4.9 g, 16.33 mmol, 1.0 equiv.) in THF (50 mL) was added potassium tert-butoxide (1.8 g, 16.33 mmol, 1.0 equiv.) at −78° C., and the reaction was stirred for 1 h. Upon completion, the reaction mixture was transferred to DM water (180 mL), and the product was extracted with ethyl acetate (4×50 mL). The organic layers were combined, washed with brine solution (150 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography; the compound was eluted with 20% ethyl acetate in hexane to give PA9.3 (1.3 g, 36.32%) as an oily white solid. MS (ES): m / z 220.1 [M+H] +

[0450] Step-4. 2,2-Dimethylmorpholin-3-one (PA9.4) To a solution of 4-benzyl-2,2-dimethylmorpholin-3-one (PA9.3) (1.3 g, 5.93 mmol, 1.0 equiv.) in toluene (25 mL) was added triflic acid (2 mL) at room temperature. The reaction mixture was stirred at 200° C. for 20 minutes under microwave irradiation. Upon completion, the reaction mixture was basified with ammonia in methanol and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, and the compound was eluted with 2.0% methanol in dichloromethane to give PA9.4 (0.5 g, 65.30%). MS (ES): m / z 130.08 [M+H] +

[0451] Step-5. 4-(6-chloropyridin-3-yl)-2,2-dimethylmorpholin-3-one (PA9.6) A solution of 2,2-dimethylmorpholin-3-one (PA9.4) (1.5 g, 11.62 mmol, 1.0 equiv.), 5-bromo-2-chloropyridine (PA9.5) (2.23 g, 11.62 mmol, 1.0 equiv.), potassium phosphate tribasic (7.3 g, 34.86 mmol, 3.0 equiv.), and 1,2-dimethylethylenediamine (0.409 g, 4.64 mmol, 0.4 equiv.) in 1,4-dioxane (15 mL) was degassed for 15-20 min. Copper(I) iodide (0.442 g, 2.32 mmol, 0.2 equiv.) was added to the reaction mixture and stirred at 110 °C for 2 h. After completion, the reaction mixture was transferred to DM water (150 mL) and the product was extracted with ethyl acetate (3 × 60 mL). The organic layers were combined, washed with brine solution (200 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material, which was further purified by column chromatography; the compound was eluted with 16% ethyl acetate in hexane to give PA9.6 (1.0 g, 35.78%). MS (ES): m / z 241.07 [M+H] +

[0452] Step-6. 2-Chloro-5-(2,2-dimethyl-3-oxomorpholino)pyridine 1-oxide (PA9.7) To a solution of 4-(6-chloropyridin-3-yl)-2,2-dimethylmorpholin-3-one (PA9.6) (1.0 g, 4.16 mmol, 2.0 equiv.) in DCM (10 mL), hydrogen peroxide-urea (CAS no. 124-43-6) (2.3 g, 24.96 mmol, 6.0 equiv.) was added slowly in small portions, and trifluoroacetic anhydride (3.5 mL, 24.96 mmol, 6.0 equiv.) was added at 0 °C. The reaction was stirred at room temperature for 1 h. Upon completion, the reaction mixture was transferred to saturated sodium sulfite solution (150 mL), and the product was extracted with dichloromethane (3 × 70 mL). The organic layers were combined, washed with brine solution (150 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material, which was used in the next step without purification. PA9.7 (1.3 g crude, quant. %) MS(ES): m / z 257.06[M+H] +

[0453] Step-7. 6-Chloro-3-(2,2-dimethyl-3-oxomorpholino)picolinonitrile (PA9) To a solution of 2-chloro-5-(2,2-dimethyl-3-oxomorpholino)pyridine 1-oxide (PA9.7) (1.3 g, 5.058 mmol, 2.0 equiv.) in acetonitrile (15 mL), triethylamine (2.1 mL, 15.17 mmol, 3.0 equiv.) and trimethylsilyl cyanide (1.5 g, 15.17 mmol, 3.0 equiv.) were added at room temperature. The reaction was stirred at 110 °C for 16 h. Upon completion, the reaction mixture was transferred to water (90 mL), and the product was extracted with ethyl acetate (3 × 40 mL). The organic layers were combined, washed with brine solution (80 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography; the compound was eluted with 10–12% ethyl acetate in hexane to give PA9 (0.7 g, 52.02%). MS(ES): m / z 266.07[M+H] +

[0454] Method PA10 - Preparation of 6-chloro-3-(2,2-dimethyl-3-oxomorpholino)picolinonitrile (PA10) [ka] Step-1. tert-Butyl (5-(3,6-dihydro-2H-pyran-4-yl)-6-(1-(dimethylamino)ethyl)pyridin-2-yl)carbamate (10.1) A solution of tert-butyl (5-bromo-6-(1-(dimethylamino)ethyl)pyridin-2-yl)carbamate (10.0) (6 g, 17.44 mmol, 1.0 equiv.) and 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (5.4 g, 26.16 mmol, 1.5 equiv.), and potassium phosphate tribasic (11.0 g, 52.32 mmol, 3.0 equiv.) in 1,4-dioxane:water (48 mL:12 mL) was degassed under a stream of N. After 15 min, chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (CAS No. 1310584-14-5) (1.3 g, 1.74 mmol, 0.1 equiv.) was added, and the reaction was stirred at 120 °C for 20 min. Upon completion, the reaction mixture was diluted with water (250 mL) and extracted with ethyl acetate (3 × 120 mL). The combined organic extracts were washed with brine (150 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give the crude material. The crude residue was purified by column chromatography with a gradient elution of 3–5% methanol in dichloromethane to give 10.1 (5.2 g, 85.87%), MS (ES): m / z 348.2 [M+H]. + , was obtained.

[0455] Step-2. tert-Butyl (6-(1-(dimethylamino)ethyl)-5-(tetrahydro-2H-pyran-4-yl)pyridin-2-yl)carbamate (10.2) To a solution of tert-butyl (5-(3,6-dihydro-2H-pyran-4-yl)-6-(1-(dimethylamino)ethyl)pyridin-2-yl)carbamate (10.1) (5.2 g, 14.98 mmol, 1.0 equiv.) in methanol:THF (45 ml:10 mL) was added acetic acid (3.0 mL), followed by wet 20% palladium hydroxide on carbon (3.6 g) and ammonium formate (2.8 g, 44.94 mmol, 3.0 equiv.). Hydrogen was purged through the reaction mixture at room temperature for 8 hours. Upon completion, the reaction mixture was filtered through a bed of Celite and washed with methanol (2 x 50 ml). The filtrate was concentrated under reduced pressure to give the crude material, which was further purified by trituration with n-pentane to give 10.2 (3.0 g, 57.36%). MS(ES): m / z 350.2 [M+H] +

[0456] Step-3. 6-(1-(dimethylamino)ethyl)-5-(tetrahydro-2H-pyran-4-yl)pyridin-2-amine (PA10) tert-Butyl (6-(1-(dimethylamino)ethyl)-5-(tetrahydro-2H-pyran-4-yl)pyridin-2-yl)carbamate (10.2) (3.0 g, 8.59 mmol, 1.0 equiv) was dissolved in dichloromethane (30 mL), and trifluoroacetic acid (12 mL) was added to the reaction mixture. The reaction was stirred at room temperature for 1 hour. Upon completion, the reaction mixture was concentrated under reduced pressure to give a residue, which was diluted with water (30 mL) and dichloromethane (15 mL). The aqueous layer was collected, neutralized with 1N NaOH solution, and extracted with 10% methanol in dichloromethane (3 × 40 mL). The combined organic extracts were washed with brine (50 mL), then dried over NaSO, filtered, and concentrated under reduced pressure to give PA10 (2.0 g, 93.43%). MS (ES): m / z 250.1 [M+H] +

[0457] Method PA11 - Preparation of 6-chloro-3-(1-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)picolinonitrile (PA11) [ka] Step-1. 1-(6-chloropyridin-3-yl)ethan-1-ol (PA11.1) To a solution of 1-(6-chloropyridin-3-yl)ethan-1-one (PA11.0) (10 g, 64.51 mmol, 1.0 equiv) in methanol (100 mL) was added sodium borohydride (1.2 g, 32.25 mmol, 0.5 equiv) in small portions at room temperature and stirred for 20 minutes at room temperature. Upon completion, the reaction mixture was transferred to water (400 mL) and the product was extracted with DCM (4 x 150 mL). The organic layers were combined, washed with brine solution (200 mL), dried over sodium sulfate, and concentrated under reduced pressure to give PA11.1 (10 g, 98.72%). The crude material was used in the next step without purification. MS (ES): m / z 158.03 [M+H] +

[0458] Step 2. 5-(1-Bromoethyl)-2-chloropyridine (PA11.2) A solution of 1-(6-chloropyridin-3-yl)ethan-1-ol (11.1) (10.0 g, 63.69 mmol, 1.0 equiv) in DCM (100 mL) was cooled to 0 °C. Triphenylphosphine (25.0 g, 95.53 mmol, 1.5 equiv) was added, followed by N-bromosuccinimide (17.0 g, 95.53 mmol, 1.5 equiv) under a N atmosphere at 0 °C and stirred at room temperature for 2 h. Upon completion, the reaction mixture was concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, and the compound was eluted with 2-3% ethyl acetate in hexane to give PA11.2 (10.2 g, 72.90%). MS (ES): m / z 220.9 [M+H] +

[0459] Step-3. 2-Chloro-5-(1-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)pyridine (PA11.3) To a solution of sodium hydride (2.22 g, 92.72 mmol, 2.0 equiv) in dimethylformamide (80 mL) was added tetrahydro-2H-pyran-4-ol (7.09 g, 69.54 mmol, 1.5 equiv) dissolved in DMF slowly at −10° C. under a N atmosphere and stirred at −10° C. for 1 h. A solution of 5-(1-bromoethyl)-2-chloropy (PA11.2) (10.2 g, 46.36 mmol, 1.0 equiv) in dimethylformamide (25 mL) was added slowly at −10° C. and stirred for 30 min, followed by stirring at 50° C. for 16 h. Upon completion, the reaction mixture was transferred to water (350 mL) and the product was extracted with ethyl acetate (3×150 mL). The organic layers were combined, washed with brine solution (250 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material, which was further purified by column chromatography; the compound was eluted with 8-10% ethyl acetate in hexane to give PA11.3 (2.2 g, 19.68%). MS (ES): m / z 242.2 [M+H] +

[0460] Step-4. 2-Chloro-5-(1-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)pyridine 1-oxide (PA11.4) To a solution of 2-chloro-5-(1-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)pyridine (PA11.3) (1.2 g, 4.97 mmol, 2.0 equiv) in DCM (15 mL) was added hydrogen peroxide-urea (CAS no. 124-43-6) (2.8 g, 29.82 mmol, 6.0 equiv) slowly in small portions, followed by trifluoroacetic anhydride (4.2 mL, 29.82 mmol, 6.0 equiv) at 0 °C. The reaction was stirred at room temperature for 1 h. Upon completion, the reaction mixture was transferred to sodium sulfite solution (120 mL) and the product was extracted with dichloromethane (3 × 50 mL). The organic layers were combined, dried over sodium sulfate, and concentrated under reduced pressure to give the crude material, which was used in the next step without further purification. PA11.4 (1.7 g crude, quant. %) MS(ES): m / z 258.01[M+H] +

[0461] Step-5. 6-Chloro-3-(1-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)picolinonitrile (PA11) To a solution of 2-chloro-5-(1-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)pyridine 1-oxide (PA11.4) (1.0 g, 3.90 mmol, 2.0 equiv.) in acetonitrile (10 mL) was added triethylamine (1.6 mL, 11.7 mmol, 3.0 equiv.) and trimethylsilyl cyanide (1.1 g, 11.7 mmol, 3.0 equiv.) at room temperature and stirred at 110 °C for 16 h. Upon completion, the reaction mixture was transferred to water (100 mL) and the product was extracted with ethyl acetate (3 × 40 mL). The organic layers were combined, washed with brine solution (100 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography; the compound was eluted with 8–10% ethyl acetate in hexane to give PA11 (0.3 g, 28.99%). MS(ES): m / z 267.09[M+H] +

[0462] Method PA12 - Preparation of 6-((dimethylamino)methyl)-5-(1-(tetrahydro-2H-pyran-4-yl)ethyl)pyridin-2-amine (PA12) [ka] Step-1. 1-(6-chloropyridin-3-yl)-1-(tetrahydro-2H-pyran-4-yl)ethan-1-ol (PA12.1) To a cooled solution of 5-bromo-2-chloropyridine (PA12.0) (2 g, 10.0 mmol, 1.0 equiv) in tetrahydrofuran (40 mL) was added n-butyllithium (6.2 mL, 15.5 mmol, 1.5 equiv) at −78°C. The reaction mixture was stirred at −78°C for 15–20 min. 1-(tetrahydro-2H-pyran-4-yl)ethan-1-one (16.4 g, 94.33 mmol, 1.5 equiv) was added slowly at −78°C and stirred for 30 min. Upon completion, the reaction mixture was quenched with ice-cold water (200 mL), extracted with ethyl acetate (3 × 100 mL), and the combined organic layers were washed with brine (150 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, eluting the compound with 16% ethyl acetate in hexane to give PA112.1 (1.6 g, 63.69%) as a sticky solid. MS (ES): m / z 242.2 [M+H] +

[0463] Step-2. 2-Chloro-5-(1-(tetrahydro-4H-pyran-4-ylidene)ethyl)pyridine (PA12.2) To a solution of 1-(6-chloropyridin-3-yl)-1-(tetrahydro-2H-pyran-4-yl)ethan-1-ol (PA12.1) (13 g, 0.537 mmol, 1.0 equiv.) in triethylsilane (20 mL) was added trifluoroacetic acid (20 mL) at 95 °C, and the reaction was stirred for 24 hours. Upon completion, the reaction mixture was cooled to room temperature and poured into sodium bicarbonate solution (350 mL), and the product was extracted with 10% methanol in dichloromethane (3 x 150 mL). The combined organic layers were washed with brine solution (150 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, and the compound was eluted with 2.5% ethyl acetate in hexane to give PA12.2 (10 g, 83.12%). MS (ES): m / z 225.2 [M+H] +

[0464] Step-3. N-(5-(1-(tetrahydro-4H-pyran-4-ylidene)ethyl)pyridin-2-yl)cyclopropanecarboxamide (PA12.3) A solution of 2-chloro-5-(1-(tetrahydro-4H-pyran-4-ylidene)ethyl)pyridine (PA12.2) (1 g, 4.46 mmol, 1.0 equiv.), cyclopropylcarboxamide (0.758 g, 8.9 mmol, 2.0 equiv.), and potassium carbonate (1.8 g, 13.3 mmol, 3.0 equiv.) in 1,4-dioxane (10 mL) was degassed under a stream of N. After 15 min, Xantphos (0.576 g, 0.892 mmol, 0.2 equiv.) and Pd(dba) (0.408 g, 0.446 mmol, 0.1 equiv.) were added, and the reaction was heated to 110 °C and stirred for 2 h. The reaction mixture was cooled to room temperature and then diluted with water (50 mL) and ethyl acetate (3 x 20 mL), and the combined organic extracts were washed with brine (50 mL), then dried over NaSO, filtered, and concentrated under reduced pressure. The crude residue was purified by column chromatography to give (PA12.3) (0.520 g, 42.71%). MS (ES): m / z 273.5 [M+H] +

[0465] Step-4. N-(5-(1-(tetrahydro-2H-pyran-4-yl)ethyl)pyridin-2-yl)cyclopropanecarboxamide (PA12.4) To a solution of N-(5-(1-(tetrahydro-4H-pyran-4-ylidene)ethyl)pyridin-2-yl)cyclopropanecarboxamide (PA12.3) (7.0 g, 25.68 mmol, 1.0 equiv) in methanol (30 mL) was added 20% wet palladium hydroxide on carbon (4 g). The reaction mixture was stirred at room temperature under an atmosphere of hydrogen gas for 24 hours. Upon completion, the reaction mixture was filtered through a pad of Celite and the filtrate was concentrated under reduced pressure to afford the crude residue PA12.4 (5 g, 70.90%). The crude material was used in the next step without purification. MS (ES): m / z 275.1 [M+H] +1

[0466] Step-5. N-(5-(1-(tetrahydro-2H-pyran-4-yl)ethyl)pyridin-2-yl)cyclopropanecarboxamide (PA12.5) A solution of N-(5-(1-(tetrahydro-2H-pyran-4-yl)ethyl)pyridin-2-yl)cyclopropanecarboxamide (PA12.4) (5 g, 18.24 mmol, 1.0 equiv) in dichloromethane (50 mL) was cooled to 0 °C, and 3-chlorobenzoic acid (18.89 g, 109.4 mmol, 6.0 equiv) was added. The reaction was stirred at room temperature for 16 h, then quenched in water (100 mL) and extracted with dichloromethane (3 × 50 mL). The combined organic extracts were washed with brine (50 mL), then dried over NaSO, filtered, and concentrated under reduced pressure to give the crude material (4 g, N-oxide), which was used in the next step. To the crude material (3 g, 10.33 mmol, 1.0 equiv.) were added nitroethane (30 mL), cyclopropylcarbonyl chloride (9.21 g, 82.65 mmol, 8.0 equiv.), and trimethylsilyl cyanide (8.19 g, 82.65 mmol, 8.0 equiv.). The reaction mixture was stirred at 50 °C for 4 h. Upon completion, the reaction mixture was quenched in water (60 mL) and extracted with dichloromethane (3 × 20 mL). The combined organic extracts were washed with brine (50 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give the crude material. The crude residue was purified by column chromatography eluting with 32% ethyl acetate in hexane to give PA12.5 (2.5 g, 45.82%) as a yellowish solid. MS (ES): m / z 300 [M+H] +

[0467] Step-6. N-(6-(aminomethyl)-5-(1-(tetrahydro-2H-pyran-4-yl)ethyl)pyridin-2-yl)cyclopropanecarboxamide (PA12.6) To a solution of N-(5-(1-(tetrahydro-2H-pyran-4-yl)ethyl)pyridin-2-yl)cyclopropanecarboxamide (PA12.5) (0.400 g, 1.088 mmol, 1.0 equiv) in methanol (2 mL) and tetrahydrofuran (2 ml) was added hydrazine hydride (1 ml) and Raney nickel (0.200 g), and the reaction was stirred at room temperature for 2 hours. Upon completion, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (3 × 20 mL). The combined organic extracts were washed with brine (50 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The crude residue was purified by column chromatography eluting with 45% ethyl acetate in hexane to give PA12.6 (0.200 g, 49.34%). MS (ES): m / z 304 [M+H] + (Note: Six parallel batches were planned.)

[0468] Step-7. N-(6-((dimethylamino)methyl)-5-(1-(tetrahydro-2H-pyran-4-yl)ethyl)pyridin-2-yl)cyclopropanecarboxamide (PA12.7) To a solution of N-(6-(aminomethyl)-5-(1-(tetrahydro-2H-pyran-4-yl)ethyl)pyridin-2-yl)cyclopropanecarboxamide (PA12.6) (0.300 g, 1.088 mmol, 1.0 equiv) in methanol (4 mL) was added formaldehyde (0.148 g, 4.9 mmol, 5.0 equiv) and acetic acid (0.3 mL), and the reaction was stirred at room temperature for 1 hour. Sodium cyanoborohydride (0.186 g, 2.9 mmol, 3.0 equiv) was added, and the reaction mixture was stirred at room temperature for 30 minutes. Upon completion, the reaction mixture was diluted with water (50 mL), extracted with dichloromethane (3 × 20 mL), and the combined organic extracts were washed with brine (50 mL), then dried over NaSO, filtered, and concentrated under reduced pressure. The crude residue was purified by column chromatography eluting with 55% ethyl acetate in hexane to give (PA12.7) (0.150 g, 45.77%). MS (ES): m / z 372.4 [M+H] +(Note: Four parallel batches were planned.)

[0469] Step-8. 6-((Dimethylamino)methyl)-5-(1-(tetrahydro-2H-pyran-4-yl)ethyl)pyridin-2-amine (PA12) To a solution of N-(6-((dimethylamino)methyl)-5-(1-(tetrahydro-2H-pyran-4-yl)ethyl)pyridin-2-yl)cyclopropanecarboxamide (PA12.70) (0.650 g, 1.96 mmol, 1.0 equiv) in methanol (8 mL) was added sodium hydroxide (1.96 g, 49.0 mmol, 25.0 equiv). The reaction was stirred at 70 °C for 1 h. After completion, the reaction mixture was diluted with water (30 mL) and extracted with dichloromethane (3 × 20 mL). The combined organic extracts were washed with brine (10 mL), dried over NaSO, and concentrated under reduced pressure. The crude residue was purified by column chromatography eluting with 5% methanol in DCM to give PA12 (0.150 g, 55.17%). MS (ES): m / z 264.4 [M+H] +

[0470] Method PA13 - Preparation of 1-(6-bromo-3-((4,4-difluorocyclohexyl)methoxy)pyridin-2-yl)-N,N-dimethylmethanamine (PA13) [ka] Step 1. (4,4-Difluorocyclohexyl)methyl 4-methylbenzenesulfonate (PA13.1) To a solution of (4,4-difluorocyclohexyl)methanol (PA13.0) (2.0 g, 13.33 mmol, 1.0 equiv) in DCM (20 mL) was added triethylamine (2.8 mL, 19.99 mmol, 1.5 equiv), followed by 4-dimethylaminopyridine (0.487 g, 3.99 mmol, 0.3 equiv). The reaction was cooled to 0 °C, and 4-toluenesulfonyl chloride (3.0 g, 15.99 mmol, 1.2 equiv) was added and stirred at 60 °C for 3 h. Upon completion, the reaction mixture was transferred to water (120 mL), and the product was extracted with dichloromethane (3 × 50 mL). The organic layers were combined, washed with brine solution (100 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the crude material. This was further purified by column chromatography, and the compound was eluted with 15-20% ethyl acetate in hexane to give PA13.1 (2.7 g, 66.61%). MS (ES): m / z 305.2 [M+H] +

[0471] Step-2. 1-(6-bromo-3-((4,4-difluorocyclohexyl)methoxy)pyridin-2-yl)-N,N-dimethylmethanamine (13) To a solution of 6-bromo-2-((dimethylamino)methyl)pyridin-3-ol (PA13.2) (1.0 g, 4.32 mmol, 1.0 equiv.) and (4,4-difluorocyclohexyl)methyl 4-methylbenzenesulfonate (PA13.1) (1.97 g, 6.49 mmol, 1.5 equiv.) in dimethylformamide (10 mL) was added potassium carbonate (1.78 g, 12.96 mmol, 3.0 equiv.). The reaction mixture was stirred at 100 °C for 16 h. Upon completion, the reaction mixture was transferred to ice-cold water (130 mL) and the product was extracted with ethyl acetate (2 × 80 mL). The organic layers were combined, washed with brine solution (100 mL), dried over sodium sulfate, and concentrated under reduced pressure to give the cru...

Claims

1. A compound of formula I: 【Chemical 77】 or a pharmaceutically acceptable salt thereof, wherein: Z is CR or N; X is —NR—; R １ teeth, 【Chemistry 86】 【Hua 87】 【Hua 88】 【Chemistry 89】 and R ２ is a 6-11 membered saturated, partially unsaturated or unsaturated fused, bridged or spiro bicyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, each of which is selected from R at an instance of q. ｃ is replaced by 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 selected from 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 NR 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; naphthyl; 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; an optionally substituted group 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 spirocyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or a 6-11 membered saturated or partially unsaturated bicyclic heterocyclic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, each of which is independently selected from R at an instance of r and R at an instance of s. Ｄ is replaced by R Ｄ Each instance of is independently selected from 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 NR 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; naphthyl; 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; an optionally substituted group selected from: a 7-12 membered saturated or partially unsaturated bicyclic heterocycle having a heteroatom; 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; or 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; 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; and A compound of Formula I, or a pharmaceutically acceptable salt thereof, wherein each s is independently 0, 1, 2, 3, or 4.

2. The compound of claim 1 , wherein X is —NH—.

3. R １ teeth, 【Hua 79-1】 【Chemistry 79-2】 【Hua 79-3】 3. The compound of claim 1 or claim 2, wherein:

4. R ２ is a 7-10 membered fused bicyclic ring having 1-3 nitrogen atoms, each of which is an instance of R Ｃ 10. The compound of claim 1, wherein each of the groups is substituted with:

5. R ２ teeth, 【Chemistry 80】 2. The compound of claim 1, wherein:

6. That R Ｃ R together with the substituent ２ teeth, 【Chemistry 81】 2. The compound of claim 1, wherein:

7. R ３ The compound of claim 1 , wherein is hydrogen.

8. R Ｃ Each instance of is independently methyl, fluoro, methoxy, -CHF ２ , 【Chemistry 82】 2. The compound of claim 1, wherein:

9. R Ｃ Each instance of is independently methyl, fluoro, methoxy, 【Chemistry 83】 2. The compound of claim 1, wherein:

10. R Ｄ Each instance of may independently be hydroxy, fluoro, methoxy, 【Chemistry 84】 2. The compound of claim 1, wherein:

11. R Ｄ Each instance of may independently be hydroxy, fluoro, methoxy, or 【Chemistry 85】 2. The compound of claim 1, wherein:

12. 2. The compound of claim 1, wherein the compound is selected from those shown in Table 1, or a pharmaceutically acceptable salt thereof.

13. 10. A pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

14. 14. A composition comprising a compound according to claim 1 or a pharmaceutical composition according to claim 13 for use as a medicament.

15. 14. A composition comprising the compound of claim 1, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 13, for use in a method of inhibiting HPK1 in a biological sample, the method comprising contacting the sample with the compound, or a pharmaceutically acceptable salt thereof, or with the pharmaceutical composition.

16. 14. A composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 13, for treating an HPK1-mediated disorder, disease, or condition in a patient.

17. 17. The composition or pharmaceutical composition of claim 16, wherein the disorder is a proliferative disorder.

18. 18. The composition or pharmaceutical composition of claim 17, wherein the proliferative disorder is cancer.

19. 18. The composition or pharmaceutical composition of claim 17, wherein the proliferative disorder is associated with one or more activating mutations in HPK1.

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