USP1 inhibitor and application thereof

Abstract

Provided herein are inhibitors of ubiquitin-specific processing protease 1 (USP1), pharmaceutical compositions thereof, and methods of use for the treatment of diseases or conditions, such as cancers described herein.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 367,268, filed June 29, 2022, the entire contents of which are incorporated herein by reference in their entirety. Technical Field

[0003] The present invention relates to compounds and methods useful for inhibiting ubiquitin-specific processing protease 1 (USP1). The present invention also provides pharmaceutically acceptable compositions comprising the compounds of the present invention and methods of using the compositions to treat various diseases, disorders, and conditions as described herein. Background Art

[0004] USP1 is a cysteine ​​protease belonging to the DUB family. It deubiquitinates a variety of cellular targets that play a role in DDR pathways, immune responses, and cancer. USP1 interacts with UAF1 (USP1-associated factor 1) to form a stable complex required for deubiquitinase activity. Well-characterized substrates of the USP1 / UAF1 complex, such as PCNA (proliferating cell nuclear antigen) and FANCD2 (Fanconi anemia complementation group D2), are key players in the processes of DNA translesion synthesis (TLS) and interstrand crosslink (ICL) repair, respectively. These two pathways are crucial for repairing DNA damage caused by DNA crosslinking agents such as cisplatin and mitomycin C. Therefore, USP1, through deubiquitination of Fanconi anemia proteins and PCNA, is a key regulator of genomic integrity and a potential synthetic lethal drug target for cancer cells that often harbor defects in certain DDR pathways. Summary of the Invention

[0005] It has now been found that the compounds of the present invention and pharmaceutically acceptable compositions thereof are effective as USP1 inhibitors. In one aspect, the present invention provides a compound of formula I:

[0006]

[0007] or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined herein and as described in the Examples herein.

[0008] The compounds of the present invention, and pharmaceutically acceptable salts and compositions thereof, can be used to treat a variety of diseases, disorders or conditions associated with USP 1. Such diseases, disorders or conditions include cell proliferative disorders (e.g., cancer as described herein). BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 Tumor volume changes after treatment are depicted.

[0010] Figure 2 Weight changes after treatment are depicted. DETAILED DESCRIPTION

[0011] 1. General Description of Certain Embodiments of the Invention:

[0012] The compounds of the present invention and their pharmaceutical salts and compositions are useful as inhibitors of USP1. Without wishing to be bound by any particular theory, it is believed that the compounds of the present invention and their pharmaceutical compositions inhibit the activity of USP1 and thus treat diseases, disorders or conditions associated with USP1, such as cancer.

[0013] In one aspect, the present invention provides a compound of formula I:

[0014]

[0015] or a pharmaceutically acceptable salt thereof, wherein:

[0016] is a single bond or a double bond;

[0017] X 1 O, N, N(R 11 )、C(O)、CR 12 or C(R 12 )2;

[0018] X 2 O, N, N(R 11 )、C(O)、CR 12 or C(R 12 )2;

[0019] X 3 N(R 11 ), C(O) or C(R 12 )2;

[0020] X 4 N or CR 12 ;

[0021] X 5 N or CR 12 ;

[0022] Each R 11 are independently hydrogen, deuterium, R or -C(O)-R;

[0023] Each R 12 are independently hydrogen, deuterium, halogen, R, -OR, -NHR, -N(R)2, -C(O)-R, -COOR, -C(O)-NHR, or -C(O)-N(R)2;

[0024] Ring A is a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S;

[0025] Each R 1 are independently halogen, R, -OR, -NHR, -N(R)2, -C(O)-R, -C(O)-NHR, or -C(O)-N(R)2;

[0026] Ring B is selected from a benzene ring, a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S, or a 5- to 6-membered heterocyclyl ring having 1 to 4 heteroatoms selected from N, O and S;

[0027] Each R 2 are independently halogen, R, -OR, -NHR, -N(R)2 or -C(O)-R;

[0028] R 3 is optionally substituted phenyl, or optionally substituted 5- to 6-membered heteroaryl having 1 to 4 heteroatoms selected from N, O and S;

[0029] R 4 and R 5 Each of is independently hydrogen, deuterium, halogen, R, -OR, -NHR or -N(R)2; or

[0030] R 4 and R 5 together with the atoms to which they are attached, form an optionally substituted ring selected from a C3-C7 carbocycle or a 3- to 7-membered heterocycle having 1 to 4 heteroatoms selected from N, O and S;

[0031] Each R is independently an optionally substituted C 1-6 an aliphatic, or an optionally substituted ring selected from a benzene ring, a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S, a C3-C7 carbocyclic ring, or a 3- to 7-membered heterocyclic ring having 1 to 4 heteroatoms selected from N, O or S;

[0032] n is 0, 1, 2, or 3; and

[0033] m is 0, 1, 2 or 3.

[0034] 2. Compounds and Definitions:

[0035] The compounds of the present invention include compounds generally described herein and are further described by the categories, subclasses and species disclosed herein. Unless otherwise indicated, as used herein, the following definitions shall apply. For the purposes of the present invention, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th edition. In addition, the general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5th edition, editors: Smith, MB and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are hereby incorporated by reference.

[0036] As used herein, the term "aliphatic" or "aliphatic group" means a straight (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more unsaturated units, or a monocyclic or bicyclic hydrocarbon (also referred to herein as "carbocycle," "cycloaliphatic," or "cycloalkyl") that is fully saturated or contains one or more unsaturated units but is not aromatic, having a single point of attachment to the rest of the molecule. Unless otherwise specified, an aliphatic group contains 1 to 6 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1 to 5 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1 to 4 aliphatic carbon atoms. In still other embodiments, an aliphatic group contains 1 to 3 aliphatic carbon atoms, and in yet other embodiments, an aliphatic group contains 1 to 2 aliphatic carbon atoms. In some embodiments, "cycloaliphatic" (or "carbocycle" or "cycloalkyl") refers to a monocyclic C3-C6 hydrocarbon that is fully saturated or contains one or more unsaturated units 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, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups, and mixtures thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl.

[0037] As used herein, the term "bicyclic" or "bicyclic ring system" refers to any bicyclic ring system, i.e., carbocyclic or heterocyclic, saturated or with one or more unsaturated units, with one or more common atoms between the two rings of the ring system. Therefore, the term includes any allowed ring fusion, such as ortho-fused or spirocyclic. As used herein, the term "heterobicyclic" is a subset of "bicyclic", which requires that one or more heteroatoms are present in one or both rings of the bicyclic ring. Such heteroatoms may be present at the ring junction and are optionally substituted and may be selected from nitrogen (including N-oxides), oxygen, sulfur (including oxidized forms, such as sulfones and sulfonates), phosphorus (including oxidized forms, such as phosphates), boron, etc. In some embodiments, the bicyclic group has 7 to 12 ring members and 0 to 4 heteroatoms independently selected from nitrogen, oxygen or sulfur. As used herein, the term "bridged bicyclic" refers to any bicyclic ring system, i.e., carbocyclic or heterocyclic, saturated or partially unsaturated, with at least one bridge. As defined by IUPAC, "bridge" is an unbranched atomic chain or an atom or valence bond connecting two bridgeheads, wherein "bridgehead" is any backbone atom combined with three or more backbone atoms (excluding hydrogen) of a ring system. In certain embodiments, the bridged bicyclic group has 7-12 ring members and 0-4 heteroatoms independently selected from nitrogen, oxygen or sulphur. Such bridged bicyclic groups are well known in the art and include those groups shown below, wherein each group is connected to the remainder of the molecule at any substitutable carbon or nitrogen atom. Unless otherwise specified, the bridged bicyclic group is optionally substituted by one or more substituents shown in the aliphatic group. Additionally or alternatively, any substitutable nitrogen of the bridged bicyclic group is optionally substituted. Exemplary bicyclic groups include:

[0038]

[0039] Exemplary bridged double rings include:

[0040]

[0041] The term "lower alkyl" refers to a C 1-4 Straight or branched chain alkyl groups. Exemplary lower alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl and tert-butyl.

[0042] The term "lower haloalkyl" refers to a C 1-4 Straight-chain or branched alkyl groups.

[0043] The term "heteroatom" means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including any oxidized form of nitrogen, sulfur, phosphorus, or silicon; a quaternized form of any basic nitrogen; or a substitutable nitrogen of a heterocyclic ring, such as N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR + (as in N-substituted pyrrolidinyl)).

[0044] As used herein, the term "unsaturated" means that a moiety has one or more units of unsaturation.

[0045] As used herein, the term "divalent C 1-8 (or C 1-6 ) saturated or unsaturated straight or branched hydrocarbon chain” refers to straight or branched divalent alkylene, alkenylene and alkynylene chains as defined herein.

[0046] The term "alkylene" refers to a divalent alkyl group. An "alkylene chain" is a polymethylene group, i.e., -(CH2) n -, wherein n is a positive integer, preferably 1 to 6, 1 to 4, 1 to 3, 1 to 2 or 2 to 3. A substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced by a substituent. Suitable substituents include those described below for substituted aliphatic groups.

[0047] 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 are replaced by a substituent. Suitable substituents include those described below for substituted aliphatic groups.

[0048] As used herein, the term "cyclopropenyl" refers to a divalent cyclopropyl radical having the following structure:

[0049] The term "halogen" means F, Cl, Br or I.

[0050] The term "aryl", used alone or as part of a larger moiety as in "aralkyl", "aralkoxy", or "aryloxyalkyl", refers to a monocyclic or bicyclic ring system having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains 3 to 7 ring members. The term "aryl" can 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, anthracenyl, etc., which may carry one or more substituents. As used herein, the term "aryl" also includes within its scope groups in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthalimidyl, phenanthridinyl, or tetrahydronaphthyl, etc.

[0051] 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); sharing 6, 10, or 14 pi electrons in a cyclic array; and having one to five heteroatoms in addition to carbon atoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, as well as any quaternized form of a basic nitrogen. Heteroaryl includes, but is not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. As used herein, the terms "heteroaryl" and "heteroaryl-" also include groups in which a heteroaromatic ring is fused to one or more aryl, alicyclic or heterocyclic rings, wherein the group or point of attachment is located on the heteroaromatic ring. Non-limiting examples include: indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzothiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolyl, tetrahydroisoquinolyl and pyrido [2,3-b] -1,4-oxazin-3 (4H) -one. The heteroaryl group can be monocyclic or bicyclic. The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic," any of which encompasses an optionally substituted ring. The term "heteroaralkyl" refers to an alkyl group substituted with a heteroaryl, wherein the alkyl and heteroaryl portions are independently optionally substituted.

[0052] As used herein, the terms "heterocycle," "heterocyclyl," "heterocyclyl group," 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 saturated or partially unsaturated and has, in addition to carbon atoms, one or more, preferably one to four, heteroatoms as defined above. The term "nitrogen," when used with respect to a ring atom of a heterocycle, includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0 to 3 heteroatoms selected from oxygen, sulfur, or nitrogen, nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or + NR (as in N-substituted pyrrolidinyl).

[0053] The heterocyclic ring may be attached to its side group at any heteroatom or carbon atom to produce a stable structure, and any ring atom may be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxane, dioxolane, diazepine, oxadiazolyl ... Oxazolin Thiazepine 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 rings, heteroaryl rings, or alicyclic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl. The heterocyclyl group may be monocyclic or bicyclic. The term "heterocycloalkyl" refers to an alkyl group substituted with a heterocyclyl, wherein the alkyl and heterocyclyl moieties are independently optionally substituted.

[0054] As used herein, the term "partially unsaturated" refers to a ring moiety containing at least one double or triple bond. The term "partially unsaturated" is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties, as defined herein.

[0055] As described herein, the compounds of the present invention may contain "optionally substituted" moieties. In general, the term "substituted," whether preceded by or without the term "optionally," means that one or more hydrogens of the designated 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 when more than one position in any given structure may be substituted with one or more substituents selected from a designated group, the substituents may be the same or different at each position. The combinations of substituents contemplated by the present invention are preferably those that result in the formation of stable or chemically feasible compounds. The term "stable," as used herein, refers to a compound that does not substantially change when subjected to conditions that allow its production, detection, and in certain embodiments, its recovery, purification, and use for one or more purposes disclosed herein.

[0056] Each optional substituent on a substitutable carbon is a monovalent substituent independently selected from: halogen; -(CH2); 0–4 R o ; –(CH2) 0–4 OR o ;-O(CH2) 0-4 Ro 、–O–(CH2) 0–4 C(O)OR o ; –(CH2) 0–4 CH(OR o )2;–(CH2) 0– 4SR o ; –(CH2) 0–4 Ph, which can be o Substitution; –(CH2) 0–4 O(CH2) 0–1 Ph, which can be o Substitution; –CH=CHPh, which can be R o Substitution; –(CH2) 0–4 O(CH2) 0–1 -pyridyl, which may be R o Substitution; –NO2; –CN; –N3; -(CH2) 0–4 N(R o )2;–(CH2) 0–4 N(R o )C(O)R o ;–N(R o )C(S)R o ; –(CH2) 0–4 N(R o )C(O)NR o 2;-N(R o )C(S)NR o 2;–(CH2) 0–4 N(R o )C(O)OR o ;–N(R o )N(R o )C(O)R o ;-N(R o )N(R o )C(O)NR o 2;-N(R o )N(R o )C(O)OR o ; –(CH2) 0–4 C(O)R o ;–C(S)R o ; –(CH2) 0–4 C(O)OR o ; –(CH2) 0–4 C(O)SR o ; -(CH2) 0–4 C(O)OSiR o 3;–(CH2) 0–4 OC(O)R o; –OC(O)(CH2) 0–4 SR–、SC(S)SR o ; –(CH2) 0–4 SC(O)R o ; –(CH2) 0–4 C(O)NR o 2;–C(S)NR o 2;–C(S)SR o ;–SC(S)SR o 、-(CH2) 0–4 OC(O)NR o 2;-C(O)N(OR o )R o ;–C(O)C(O)R o ;–C(O)CH2C(O)R o ;–C(NOR o )R o ; -(CH2) 0–4 SSR o ; -(CH2) 0–4 S(O)2R o ; –(CH2) 0–4 S(O)2OR o ; –(CH2) 0–4 OS(O)2R o ;–S(O)2NR o 2;-S(O)(NR o )R o ; –S(O)2N=C(NR o 2)2;-(CH2) 0–4 S(O)R o ;-N(R o )S(O)2NR o 2;–N(R o )S(O)2R o ;–N(OR o )R o ;–C(NH)NR o 2;–P(O)2R o ;-P(O)R o 2;-OP(O)R o 2;–OP(O)(OR o )2;SiR o 3;–(C 1–4 linear or branched alkylene)O–N(R o )2; or –(C 1–4 linear or branched alkylene) C(O)O–N(R o )2.

[0057] Each R o are independently hydrogen, C 1-6 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, -CH2-(5- to 6-membered heteroaryl ring) or a 5- to 6-membered saturated, partially unsaturated or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or, notwithstanding the above definitions, two independent occurrences of R o Together with their intervening atoms, they form a 3- to 12-membered saturated, partially unsaturated or aromatic monocyclic or bicyclic ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen or sulfur, which may be in an R group selected from =O and =S. o The saturated carbon atom is substituted by a divalent substituent; or each R o Optionally substituted with monovalent substituents independently selected from halogen, -(CH2) 0–2 R ● 、–(halogenated R ● ),–(CH2) 0–2 OH, –(CH2) 0–2 OR ● 、–(CH2) 0–2 CH(OR ● )2;-O(halogenated R ● ), –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 ● 、–(C 1–4 linear or branched alkylene)C(O)OR ● or –SSR ● .

[0058] Each R ● Independently selected from C 1-4 Aliphatic, –CH2Ph, –O(CH2) 0–1Ph or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and wherein each R ● is unsubstituted or, if preceded by halogen, substituted only with one or more halogens; or wherein the optional substituents on the saturated carbon are independently selected from ═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 The divalent substituent of S-, or the divalent substituent attached to an ortho-substitutable carbon of an "optionally substituted" group is -O(CR * 2) 2–3 O–, where each independent occurrence of R * Selected from hydrogen, C 1–6 an aliphatic or unsubstituted 5- to 6-membered saturated, partially unsaturated or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0059] When R * C 1–6 When aliphatic, R * Optionally halogen, –R ● 、-(halogenated R ● ),-OH, –OR ● 、–O(halogenated R ● ), –CN, –C(O)OH, –C(O)OR ● , –NH2, –NHR ● ,–NR ● 2 or –NO2, where each R ● Independently selected from C 1–4 Aliphatic, –CH2Ph, –O(CH2) 0–1 Ph or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and wherein each R ● is unsubstituted or, where preceded by halogen, substituted only by one or more halogens.

[0060] The optional substituents on the substitutable nitrogen are independently or Each of these are independently hydrogen, C 1–6aliphatic, unsubstituted -OPh, or an unsubstituted 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or two independent occurrences of Together with their intervening atoms, they form an unsubstituted 3- to 12-membered saturated, partially unsaturated or aromatic monocyclic or bicyclic ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen or sulfur; wherein when C 1–6 When aliphatic, Optionally halogen, –R ● 、-(halogenated R ● ),-OH, –OR ● 、–O(halogenated R ● ), –CN, –C(O)OH, –C(O)OR ● , –NH2, –NHR ● ,–NR ● 2 or –NO2, where each R ● Independently selected from C 1–4 Aliphatic, –CH2Ph, –O(CH2) 0–1 Ph or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and wherein each R ● is unsubstituted or, if preceded by halogen, substituted only by one or more halogens.

[0061] As used herein, the term "pharmaceutically acceptable salt" refers to salts that are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, etc., within the scope of sound medical judgment and commensurate with a reasonable benefit-risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SM 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 formed of an amino group with an inorganic acid (such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid) or with an organic acid (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, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like.

[0062] Salts derived from appropriate bases include alkali metal salts, alkaline earth metal salts, ammonium salts and N + (C 1-4 Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Where appropriate, additional pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates.

[0063] 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 for each asymmetric 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 present invention. Unless otherwise stated, all tautomeric forms of the present compounds are within the scope of the present invention. In addition, 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, a structure having the present structure (including replacement of hydrogen by deuterium or tritium, or by 13 C or 14 Compounds containing deuterium (C-enriched carbon substituted carbon) are within the scope of the present invention. Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents according to the present invention. In certain embodiments, the warhead portion of the provided compounds comprises one or more deuterium atoms.

[0064] As used herein, the term "inhibitor" or "USP1 inhibitor" or "USP1 antagonist" is defined as a compound that binds to and / or inhibits USP1 with measurable affinity. In some embodiments, inhibition in the presence of the inhibitor is observed in a dose-dependent manner. In some embodiments, the measured signal (e.g., signaling activity or biological activity) is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% lower than the signal measured with a negative control under comparable conditions. The potency of an inhibitor is generally measured by its IC 50 The IC value is defined as the half-maximal inhibitory concentration or the concentration required to inhibit the agonist response by 50%. 50 The lower the value, the more potent the antagonist is and the lower the concentration required to inhibit the maximal biological response. In certain embodiments, the inhibitor has an IC of less than about 100 μM, less than about 50 μM, less than about 1 μM, less than about 500 nM, less than about 100 nM, less than about 10 nM, or less than about 1 nM. 50 and / or binding constants.

[0065] As used herein, the terms "measurable affinity" and "measurably inhibit" mean a measurable change or inhibition of USP1 activity between a sample comprising a compound of the invention or composition thereof and USP1 and an equivalent sample comprising USP1 in the absence of the compound or composition thereof.

[0066] 3. Description of exemplary embodiments:

[0067] In one aspect, the present invention provides a compound of formula I:

[0068]

[0069] or a pharmaceutically acceptable salt thereof, wherein:

[0070] is a single bond or a double bond;

[0071] X 1 O, N, N(R 11 )、C(O)、CR 12 or C(R 12 )2;

[0072] X 2 O, N, N(R 11 )、C(O)、CR 12 or C(R 12 )2;

[0073] X 3 N(R 11 ), C(O) or C(R 12 )2;

[0074] X 4 N or CR 12 ;

[0075] X 5 N or CR 12 ;

[0076] Each R 11 are independently hydrogen, deuterium, R or -C(O)-R;

[0077] Each R 12 are independently hydrogen, deuterium, halogen, R, -OR, -NHR, -N(R)2, -C(O)-R, -COOR, -C(O)-NHR, or -C(O)-N(R)2;

[0078] Ring A is a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S;

[0079] Each R 1are independently halogen, R, -OR, -NHR, -N(R)2, -C(O)-R, -C(O)-NHR, or -C(O)-N(R)2;

[0080] Ring B is selected from a benzene ring, a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S, or a 5- to 6-membered heterocyclyl ring having 1 to 4 heteroatoms selected from N, O and S;

[0081] Each R 2 are independently halogen, R, -OR, -NHR, -N(R)2 or -C(O)-R;

[0082] R 3 is optionally substituted phenyl, or optionally substituted 5- to 6-membered heteroaryl having 1 to 4 heteroatoms selected from N, O and S;

[0083] R 4 and R 5 Each of is independently hydrogen, deuterium, halogen, R, -OR, -NHR or -N(R)2; or

[0084] R 4 and R 5 together with the atoms to which they are attached, form an optionally substituted ring selected from a C3-C7 carbocycle or a 3- to 7-membered heterocycle having 1 to 4 heteroatoms selected from N, O and S;

[0085] Each R is independently an optionally substituted C 1-6 an aliphatic, or an optionally substituted ring selected from a benzene ring, a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S, a C3-C7 carbocyclic ring, or a 3- to 7-membered heterocyclic ring having 1 to 4 heteroatoms selected from N, O or S;

[0086] n is 0, 1, 2, or 3; and

[0087] m is 0, 1, 2 or 3.

[0088] As generally defined above, Is a single bond or a double bond.

[0089] In some embodiments, is a single bond. In some embodiments, It's a double bond.

[0090] In some embodiments, Select from those depicted in Table 1 below.

[0091] As generally defined above, X 1 It is O, N, N(R 11 )、C(O)、CR12 or C(R 12 )2, wherein each variable is independently as defined herein and as described in the Examples herein.

[0092] In some embodiments, X 1 Is O. In some embodiments, X 1 is N. In some embodiments, X 1 It is N(R 11 ), where R 11 As defined herein and as described in the Examples herein. In some embodiments, X 1 In some embodiments, X 1 It's CR 12 , where R 12 As defined herein and as described in the Examples herein. In some embodiments, X 1 It is C(R 12 )2, where each R 12 As defined herein and as described in the Examples herein.

[0093] In some embodiments, X 1 Select from those depicted in Table 1 below.

[0094] As generally defined above, X 2 It is O, N, N(R 11 )、C(O)、CR 12 or C(R 12 )2, wherein each variable is independently as defined herein and as described in the Examples herein.

[0095] In some embodiments, X 2 Is O. In some embodiments, X 2 is N. In some embodiments, X 2 It is N(R 11 ), where R 11 As defined herein and as described in the Examples herein. In some embodiments, X 2 In some embodiments, X 2 It's CR 12 , where R 12 As defined herein and as described in the Examples herein. In some embodiments, X 2 It is C(R 12 )2, where each R 12 As defined herein and as described in the Examples herein.

[0096] In some embodiments, X 2 Select from those depicted in Table 1 below.

[0097] As generally defined above, X 3 It is N(R 11 ), C(O) or C(R 12 )2, wherein each variable is independently as defined herein and as described in the Examples herein.

[0098] In some embodiments, X 3 It is N(R 11 ), where R 11 As defined herein and as described in the Examples herein. In some embodiments, X 3 In some embodiments, X 3 It is C(R 12 )2, where each R 12 As defined herein and as described in the Examples herein.

[0099] In some embodiments, X 3 Select from those depicted in Table 1 below.

[0100] As generally defined above, X 4 Is N or CR 12 , where R 12 As defined herein and as described in the Examples herein.

[0101] In some embodiments, X 4 is N. In some embodiments, X 3 It's CR 12 , where R 12 As defined herein and as described in the Examples herein.

[0102] In some embodiments, X 4 Select from those depicted in Table 1 below.

[0103] As generally defined above, X 5 Is N or CR 12 , where R 12 As defined herein and as described in the Examples herein.

[0104] In some embodiments, X 5 is N. In some embodiments, X 5 It's CR 12 , where R 12 As defined herein and as described in the Examples herein.

[0105] In some embodiments, X 5 Select from those depicted in Table 1 below.

[0106] As generally defined above, each R 11R is independently hydrogen, deuterium, R, or -C(O)-R, wherein each R is independently as defined herein and as described in the Examples herein.

[0107] In some embodiments, R 11 is hydrogen. In some embodiments, R 11 In some embodiments, R 11 is R, wherein each R is independently as defined herein and as described in the Examples herein. In some embodiments, R 11 is -C(O)-R, wherein each R is independently as defined herein and as described in the Examples herein.

[0108] In some embodiments, R 11 It is -CH3 or -CD3.

[0109] In some embodiments, R 11 Select from those depicted in Table 1 below.

[0110] As generally defined above, each R 12 is independently hydrogen, deuterium, halogen, R, -OR, -NHR, -N(R)2, -C(O)-R, -COOR, -C(O)-NHR, or -C(O)-N(R)2, wherein each R is independently as defined herein and as described in the Examples herein.

[0111] In some embodiments, R 12 is hydrogen. In some embodiments, R 12 In some embodiments, R 12 In some embodiments, R 12 is R, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 12 is -OR, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 12 Is -NHR, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 12 is -N(R)2, wherein each R is independently as defined herein and as described in the Examples herein. In some embodiments, R 12 is -C(O)-R, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 12 Is -COOR, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 12 is -C(O)-NHR, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 12is -C(O)-N(R)2, wherein each R is independently as defined herein and as described in the Examples herein.

[0112] In some embodiments, R 12 is optionally substituted In some embodiments, R 12 is optionally substituted In some embodiments, R 12 is optionally substituted In some embodiments, R 12 is optionally substituted In some embodiments, R 12 is optionally substituted In some embodiments, R 12 is optionally substituted In some embodiments, R 12 is optionally substituted

[0113] In some embodiments, each R 12 are independently H, -F, -Br, -Cl, -CF3, -CHF2, -CH3, -CD3,

[0114] In some embodiments, R 12 yes In some embodiments, R 12 yes In some embodiments, R 12 yes In some embodiments, R 12 yes In some embodiments, R 12 yes In some embodiments, R 12 yes In some embodiments, R 12 yes In some embodiments, R 12 yes

[0115] In some embodiments, R 12 Select from those depicted in Table 1 below.

[0116] As generally defined above, Ring A is a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S.

[0117] In some embodiments, Ring A is a 5-membered heteroaromatic ring having 1, 2, 3, or 4 heteroatoms selected from N, O, and S. In some embodiments, Ring A is a 6-membered heteroaromatic ring having 1, 2, 3, or 4 heteroatoms selected from N, O, and S.

[0118] In some embodiments, Ring A is a 5- or 6-membered heteroaromatic ring having 1, 2, 3, or 4 heteroatoms N.

[0119] In some embodiments, Ring A is In some embodiments, Ring A is

[0120] In some embodiments, for

[0121] In some embodiments, Ring A is selected from those depicted in Table 1 below.

[0122] As generally defined above, each R 1 R is independently halogen, R, -OR, -NHR, -N(R)2, -C(O)-R, -C(O)-NHR, or -C(O)-N(R)2, wherein each R is independently as defined herein and as described in the Examples herein.

[0123] In some embodiments, R 1 In some embodiments, R 1 is R, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 1 is -OR, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 1 Is -NHR, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 1 is -N(R)2, wherein each R is independently as defined herein and as described in the Examples herein. In some embodiments, R 1 is -C(O)-R, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 1 is -C(O)-NHR, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 1 is -C(O)-N(R)2, wherein each R is independently as defined herein and as described in the Examples herein.

[0124] In some embodiments, each R 1 are independently -Cl, -F, -OCH3, -OCD3, -O-CHF2, -O-CF3 or

[0125] In some embodiments, R 1 Select from those depicted in Table 1 below.

[0126] As generally defined above, Ring B is selected from a benzene ring, a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S, or a 5- to 6-membered heterocyclyl ring having 1 to 4 heteroatoms selected from N, O and S.

[0127] In some embodiments, Ring B is a benzene ring.

[0128] In some embodiments, Ring B is a 5- to 6-membered heteroaromatic ring having 1, 2, 3, or 4 heteroatoms selected from N, O, and S. In some embodiments, Ring B is a 5-membered heteroaromatic ring having 1, 2, 3, or 4 heteroatoms selected from N, O, and S. In some embodiments, Ring B is a 6-membered heteroaromatic ring having 1, 2, 3, or 4 heteroatoms selected from N, O, and S. In some embodiments, Ring A is a 5- or 6-membered heteroaromatic ring having 1, 2, 3, or 4 heteroatoms N.

[0129] In some embodiments, Ring B is a 5- or 6-membered heterocyclyl ring having 1, 2, 3, or 4 heteroatoms selected from N, O, and S. In some embodiments, Ring B is a 5- or 6-membered heterocyclyl ring having 1, 2, 3, or 4 heteroatoms N.

[0130] In some embodiments, Ring B is

[0131] In some embodiments, Ring B is selected from those depicted in Table 1 below.

[0132] As generally defined above, each R 2 R is independently halogen, R, -OR, -NHR, -N(R)2, or -C(O)-R, wherein each R is independently as defined herein and as described in the Examples herein.

[0133] In some embodiments, R 2 In some embodiments, R 2 is R, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 2 is -OR, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 2 Is -NHR, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 2 is -N(R)2, wherein each R is independently as defined herein and as described in the Examples herein. In some embodiments, R 2 is -C(O)-R, wherein R is as defined herein and as described in the Examples herein.

[0134] In some embodiments, each R 2 is independently -F, -OCH3 or -OCD3.

[0135] In some embodiments, R 2 Select from those depicted in Table 1 below.

[0136] In some embodiments, for It is selected from In some embodiments, for It is selected from

[0137] In some embodiments, Selected from

[0138] In some embodiments, for In some embodiments, for

[0139] As generally defined above, R 3 is an optionally substituted phenyl group, or an optionally substituted 5- to 6-membered heteroaryl group having 1 to 4 heteroatoms selected from N, O and S.

[0140] In some embodiments, R 3 is optionally substituted phenyl.

[0141] In some embodiments, R 3 is an optionally substituted 5- or 6-membered heteroaryl group having 1, 2, 3, or 4 heteroatoms selected from N, O, and S. In some embodiments, R 3 is an optionally substituted 5-membered heteroaryl group having 1, 2, 3 or 4 heteroatoms selected from N, O and S. In some embodiments, R 3 is an optionally substituted 6-membered heteroaryl group having 1, 2, 3 or 4 heteroatoms selected from N, O and S. In some embodiments, R 3 is an optionally substituted 5- or 6-membered heteroaryl group having 1, 2, 3 or 4 heteroatoms N.

[0142] In some embodiments, R 3 is optionally substituted

[0143] In some embodiments, R 3 for Each R13 R is independently halogen, R or -OR, and q is 0, 1, 2 or 3.

[0144] In some embodiments, R 13 In some embodiments, R 13 is R, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 13 is -OR, wherein R is as defined herein and as described in the Examples herein.

[0145] In some embodiments, R 13 For -F, -CH3, -CD3, -CF3, -CHF2, -OCH3, -OCD3,

[0146] In some embodiments, q is 0. In some embodiments, q is 1. In some embodiments, q is 2. In some embodiments, q is 3.

[0147] In some embodiments, R 3 for

[0148] In some embodiments, R 3 for It is where R 23 is optionally substituted C 1-6 Aliphatic, and R 24 is an optionally substituted C3-C7 carbocycle. 23 -CH3, -CD3, -CF3, -CHF2, In some embodiments, R 24 yes

[0149] In some embodiments, R 3 Select from those depicted in Table 1 below.

[0150] As generally defined above, R 4 and R 5 Each of is independently hydrogen, deuterium, halogen, R, -OR, -NHR or -N(R)2; or R 4 and R 5 Together with the atoms to which they are attached, they form an optionally substituted ring selected from a C3-C7 carbocycle or a 3- to 7-membered heterocycle having 1 to 4 heteroatoms selected from N, O, and S, wherein each R is independently as defined herein and as described in the Examples herein.

[0151] In some embodiments, R 4 and R 5 is independently hydrogen, deuterium, halogen, R, -OR, -NHR, or -N(R)2, wherein each R is independently as defined herein and as described in the Examples herein.

[0152] In some embodiments, R 4 is hydrogen. In some embodiments, R 4 In some embodiments, R 4 In some embodiments, R 4 is R, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 4 is -OR, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 4 Is -NHR, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 4 is -N(R)2, wherein each R is independently as defined herein and as described in the Examples herein.

[0153] In some embodiments, R 5 is hydrogen. In some embodiments, R 5 In some embodiments, R 5 In some embodiments, R 5 is R, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 5 is -OR, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 5 Is -NHR, wherein R is as defined herein and as described in the Examples herein. In some embodiments, R 5 is -N(R)2, wherein each R is independently as defined herein and as described in the Examples herein.

[0154] In some embodiments, R 4 and R 5 Together with the atoms to which they are attached, they form an optionally substituted ring selected from a C3-C7 carbocycle or a 3- to 7-membered heterocycle having 1 to 4 heteroatoms selected from N, O, and S. In some embodiments, R 4 and R 5 Together with the atoms to which they are attached, they form an optionally substituted C3-C7 carbocycle. 4 and R 5 Together with the atoms to which they are attached, they form an optionally substituted 3-, 4-, 5-, 6-, or 7-membered heterocyclic ring having 1, 2, 3, or 4 heteroatoms selected from N, O, and S. In some embodiments, R4 and R 5 Together with the atoms to which they are attached, they form an optionally substituted 3-, 4-, 5-, 6-, or 7-membered heterocyclic ring having 1, 2, 3, or 4 heteroatoms selected from N and O. In some embodiments, R 4 and R 5 Together with the atoms to which they are attached, they form an optionally substituted 4-, 5-, or 6-membered heterocyclic ring having 1, 2, or 3 heteroatoms selected from N, O, and S. In some embodiments, R 4 and R 5 Together with the atoms to which they are attached, they form an optionally substituted 3, 4, 5, 6, or 7 membered heterocyclic ring having 1, 2, 3, or 4 heteroatoms N. In some embodiments, R 4 and R 5 Together with the atoms to which they are attached they form an optionally substituted 4-, 5- or 6-membered heterocyclic ring having 1, 2 or 3 heteroatoms N.

[0155] In some embodiments, R 4 and R 5 Each of is independently selected from those depicted in Table 1 below.

[0156] As generally defined above, each R is independently an optionally substituted C 1-6 an aliphatic, or optionally substituted ring selected from a benzene ring, a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S, a C3-C7 carbocyclic ring, or a 3- to 7-membered heterocyclic ring having 1 to 4 heteroatoms selected from N, O or S.

[0157] In some embodiments, R is optionally substituted C 1-6 In some embodiments, R is an unsubstituted C 1-6 In some embodiments, R is C substituted 1, 2, 3, 4, 5, or 6 times by halogen. 1-6 In some embodiments, R is C substituted 1, 2, 3, 4, 5, or 6 times with -F. 1-6 In some embodiments, R is -C substituted 1, 2, 3, 4, 5, or 6 times with -F. 1-3 Aliphatic. In some embodiments, R is -CH3. In some embodiments, R is -CH2CH3. In some embodiments, R is -CF3. In some embodiments, R is -CHF2.

[0158] In some embodiments, R is an optionally substituted phenyl ring.

[0159] In some embodiments, R is an optionally substituted 5- or 6-membered heteroaromatic ring having 1, 2, 3, or 4 heteroatoms selected from N, O, and S. In some embodiments, R is an optionally substituted 5- or 6-membered heteroaromatic ring having 1, 2, 3, or 4 heteroatoms selected from N, O, and S. In some embodiments, R is an optionally substituted 6-membered heteroaromatic ring having 1, 2, 3, or 4 heteroatoms selected from N, O, and S. In some embodiments, R is an optionally substituted 5- or 6-membered heteroaromatic ring having 1, 2, 3, or 4 heteroatoms N.

[0160] In some embodiments, R is an optionally substituted C3-C7 carbocycle.

[0161] In some embodiments, R is an optionally substituted 3, 4, 5, 6, or 7-membered heterocyclic ring having 1, 2, 3, or 4 heteroatoms selected from N, O, and S. In some embodiments, R is an optionally substituted 3, 4, 5, 6, or 7-membered heterocyclic ring having 1, 2, 3, or 4 heteroatoms selected from N and O. In some embodiments, R is an optionally substituted 4, 5, or 6-membered heterocyclic ring having 1, 2, or 3 heteroatoms selected from N, O, and S. In some embodiments, R is an optionally substituted 3, 4, 5, 6, or 7-membered heterocyclic ring having 1, 2, 3, or 4 heteroatoms selected from N, O, and S. In some embodiments, R is an optionally substituted 3, 4, 5, 6, or 7-membered heterocyclic ring having 1, 2, 3, or 4 heteroatoms N. In some embodiments, R is an optionally substituted 4, 5, or 6-membered heterocyclic ring having 1, 2, or 3 heteroatoms N.

[0162] In some embodiments, R is unsubstituted C 1-6 In some embodiments, R is an unsubstituted C 1-6 In some embodiments, R is C substituted 1, 2, 3, 4, 5, or 6 times with halogen or -OH. 1-6 In some embodiments, R is C substituted 1, 2, 3, 4, 5, or 6 times with halogen or -OH. 1-6 In some embodiments, R is -CH3, -CD3, -CH2F, -CHF2 or -CF3.

[0163] In some embodiments, R is optionally substituted In some embodiments, R is optionally substituted with halogen or C 1-6 Alkyl substituted 1, 2 or 3 times In some embodiments, R is optionally substituted In some embodiments, R is optionally substituted with halogen or C 1-6 Alkyl substituted 1, 2 or 3 times In some embodiments, R is optionally substituted In some embodiments, R is optionally substituted with halogen or C 1-6 Alkyl substituted 1, 2 or 3 times In some embodiments, R is optionally substituted In some embodiments, R is optionally substituted with halogen or C 1-6 Alkyl substituted 1, 2 or 3 times In some embodiments, R is optionally substituted In some embodiments, R is optionally substituted with halogen or C 1-6 Alkyl substituted 1, 2 or 3 times In some embodiments, R is optionally substituted In some embodiments, R is optionally substituted with halogen or C 1-6 Alkyl substituted 1, 2 or 3 times In some embodiments, R is optionally substituted In some embodiments, R is optionally substituted with halogen or C 1-6 Alkyl substituted 1, 2 or 3 times

[0164] In some embodiments, each R is independently -CH3, -CD3, -CF3, -CHF2,

[0165] In some embodiments, R is In some embodiments, R is In some embodiments, R is In some embodiments, R is In some embodiments, R is In some embodiments, R is In some embodiments, R is In some embodiments, R is

[0166] In some embodiments, each R is independently selected from those depicted in Table 1 below.

[0167] As generally defined above, n is 0, 1, 2 or 3.

[0168] In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0169] In some embodiments, n is selected from those depicted in Table 1 below.

[0170] As generally defined above, m is 0, 1, 2 or 3.

[0171] In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, m is 3.

[0172] In some embodiments, m is selected from those depicted in Table 1 below.

[0173] In some embodiments, the present invention provides a compound of formula II or II':

[0174]

[0175] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0176] In some embodiments, the present invention provides a compound of formula II-1 or II'-1:

[0177]

[0178] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0179] In some embodiments, the present invention provides a compound of formula II-2 or II'-2:

[0180]

[0181] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0182] In some embodiments, the present invention provides a compound of formula II-3 or II'-3:

[0183]

[0184] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0185] In some embodiments, the present invention provides a compound of formula II-4 or II'-4:

[0186]

[0187] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0188] In some embodiments, the present invention provides a compound of formula II-5:

[0189]

[0190] or a pharmaceutically acceptable salt thereof, wherein:

[0191] R2 is -OR, wherein R is optionally substituted C 1-6 aliphatic; and

[0192] Each of the remaining variables, individually and in combination, is as defined above and as described in the Examples herein.

[0193] In some embodiments, the present invention provides a compound of formula II-6, II-7, II-8 or II-9:

[0194]

[0195] or a pharmaceutically acceptable salt thereof, wherein:

[0196] R 12 is an optionally substituted 3-, 4-, 5-, 6- or 7-membered heterocyclic ring having 1, 2, 3 or 4 heteroatoms selected from N, O or S, or an optionally substituted 5- or 6-membered heteroaromatic ring having 1, 2, 3 or 4 heteroatoms selected from N, O and S; and

[0197] Each of the remaining variables, individually and in combination, is as defined above and as described in the Examples herein.

[0198] In some embodiments, the present invention provides a compound of formula II-6, II-7, II-8 or II-9:

[0199]

[0200] or a pharmaceutically acceptable salt thereof, wherein:

[0201] R 12 is an optionally substituted 5- or 6-membered heteroaromatic ring having 1, 2, 3 or 4 heteroatoms selected from N, O and S; and

[0202] Each of the remaining variables, individually and in combination, is as defined above and as described in the Examples herein.

[0203] In some embodiments, the present invention provides a compound of formula III or III':

[0204]

[0205] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0206] In some embodiments, the present invention provides a compound of formula III-1 or III'-1:

[0207]

[0208] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0209] In some embodiments, the present invention provides a compound of formula III-2 or III'-2:

[0210]

[0211] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0212] In some embodiments, the present invention provides a compound of formula III-3 or III'-3:

[0213]

[0214]

[0215] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0216] In some embodiments, the present invention provides a compound of formula III-4 or III'-4:

[0217]

[0218] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0219] In some embodiments, the present invention provides a compound of formula III-5:

[0220]

[0221] or a pharmaceutically acceptable salt thereof, wherein:

[0222] Each R 2 is independently halogen or -OR, wherein R is optionally substituted C 1-6 aliphatic;

[0223] R 23 is optionally substituted C 1-6 aliphatic;

[0224] R 24 is optionally substituted C 1-6 aliphatic, or optionally substituted C3-C7 carbocycle; and

[0225] Each of the remaining variables, individually and in combination, is as defined above and as described in the Examples herein.

[0226] In some embodiments, the present invention provides a compound of formula IV or IV':

[0227]

[0228] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0229] In some embodiments, the present invention provides a compound of formula V or V':

[0230]

[0231] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0232] In some embodiments, the present invention provides a compound of formula VI or VI':

[0233]

[0234] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0235] In some embodiments, the present invention provides a compound of formula VII or VII':

[0236]

[0237] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0238] In some embodiments, the present invention provides a compound of formula VIII or VIII':

[0239]

[0240] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0241] In some embodiments, the present invention provides a compound of Formula IX or IX':

[0242]

[0243] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0244] In some embodiments, the present invention provides a compound of formula X or X':

[0245]

[0246] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0247] In some embodiments, the present invention provides a compound of formula XI or XI':

[0248]

[0249]

[0250] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0251] In some embodiments, the present invention provides a compound of Formula XI-1 to XI-4 or XI'-1 to XI'-4:

[0252]

[0253] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0254] In some embodiments, the present invention provides a compound of Formula XII or XII':

[0255]

[0256] or a pharmaceutically acceptable salt thereof, wherein R 21 is optionally substituted C 1-6 aliphatic, and each remaining variable, individually or in combination, is as defined above and as described in the Examples herein.

[0257] In some embodiments, R 21 For unsubstituted C 1-6 In some embodiments, R 21 For unsubstituted C 1-6 In some embodiments, R 21 is C substituted 1, 2, 3, 4, 5 or 6 times by halogen or -OH 1-6 In some embodiments, R 21 is C substituted 1, 2, 3, 4, 5 or 6 times by halogen or -OH 1-6 In some embodiments, R 21 -CH3, -CD3, -CH2F, -CHF2 or -CF3.

[0258] In some embodiments, the present invention provides a compound of Formula XII-1 to XII-5 or XII'-1 to XII'-5:

[0259]

[0260]

[0261] or a pharmaceutically acceptable salt thereof, wherein R 21 is optionally substituted C 1-6 aliphatic, and each remaining variable, individually or in combination, is as defined above and as described in the Examples herein.

[0262] In some embodiments, the present invention provides a compound of formula XII-6, XII-7, XII-8, or XII-9:

[0263]

[0264]

[0265] or a pharmaceutically acceptable salt thereof, wherein:

[0266] R 12 is an optionally substituted 3-, 4-, 5-, 6- or 7-membered heterocyclic ring having 1, 2, 3 or 4 heteroatoms selected from N, O or S, or an optionally substituted 5- or 6-membered heteroaromatic ring having 1, 2, 3 or 4 heteroatoms selected from N, O and S; and

[0267] Each of the remaining variables, individually and in combination, is as defined above and as described in the Examples herein.

[0268] In some embodiments, the present invention provides a compound of Formula XII-10, XII-11, XII-12, XII-13, XII-14, or XII-15:

[0269]

[0270]

[0271] or a pharmaceutically acceptable salt thereof, wherein each variable, alone or in combination, is as defined above and as described in the Examples herein.

[0272] In some embodiments, the present invention provides a compound of Formula XIII or XIII':

[0273]

[0274] or a pharmaceutically acceptable salt thereof, wherein:

[0275] R 22 is an optionally substituted ring selected from a benzene ring and a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S; and

[0276] Each of the remaining variables, individually and in combination, is as defined above and as described in the Examples herein.

[0277] In some embodiments, the present invention provides a compound of Formula XIII-1 to XIII-5 or XIII'-1 to XIII'-5:

[0278]

[0279]

[0280] or a pharmaceutically acceptable salt thereof, wherein:

[0281] R 22 is an optionally substituted ring selected from a benzene ring and a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S; and

[0282] Each of the remaining variables, individually and in combination, is as defined above and as described in the Examples herein.

[0283] In some embodiments, R 22 is an optionally substituted phenyl group. In some embodiments, R 22 is optionally substituted with halogen or C 1-6 Alkyl substituted 1, 2 or 3 times phenyl. In some embodiments, R 22 is an optionally substituted 5- or 6-membered heteroaromatic ring having 1, 2, 3, or 4 heteroatoms selected from N, O, and S. In some embodiments, R 22 is a 5- or 6-membered heteroaromatic ring having 1, 2, 3 or 4 heteroatoms selected from N, O and S, optionally substituted by halogen or C 1-6 Alkyl is substituted 1, 2 or 3 times. In some embodiments, R 22 is optionally substituted In some embodiments, R 22 is optionally substituted with halogen or C 1-6 Alkyl substituted 1, 2 or 3 times In some embodiments, R 22 is optionally substituted In some embodiments, R 22 is optionally substituted with halogen or C 1-6 Alkyl substituted 1, 2 or 3 times In some embodiments, R 22 is optionally substituted In some embodiments, R 22 is optionally substituted with halogen or C 1-6 Alkyl substituted 1, 2 or 3 times

[0284] In some embodiments, R 22 yes In some embodiments, R 22 yes In some embodiments, R 22 yes In some embodiments, R 22 yes In some embodiments, R 22 yes In some embodiments, R 22 yes In some embodiments, R 22 yes

[0285] In some embodiments, the present invention provides a compound of Formula XIV or XIV':

[0286]

[0287] or a pharmaceutically acceptable salt thereof, wherein R 21 is optionally substituted C 1-6 aliphatic, and each remaining variable, individually or in combination, is as defined above and as described in the Examples herein.

[0288] In some embodiments, the present invention provides a compound of Formula XIV-1 to XIV-5 or XIV'-1 to XIV'-5:

[0289]

[0290]

[0291] or a pharmaceutically acceptable salt thereof, wherein R 21 is optionally substituted C 1-6 aliphatic, and each remaining variable, individually or in combination, is as defined above and as described in the Examples herein.

[0292] In some embodiments, the present invention provides a compound of formula XIV-6:

[0293]

[0294] or a pharmaceutically acceptable salt thereof, wherein:

[0295] R 12 is hydrogen, an optionally substituted 3-, 4-, 5-, 6-, or 7-membered heterocyclic ring having 1, 2, 3, or 4 heteroatoms selected from N, O, or S, or an optionally substituted 5- or 6-membered heteroaromatic ring having 1, 2, 3, or 4 heteroatoms selected from N, O, and S; and

[0296] Each of the remaining variables, individually and in combination, is as defined above and as described in the Examples herein.

[0297] Exemplary compounds of the present invention are set forth in Table 1 below.

[0298] In some embodiments, the present invention provides compounds described in Table 1 below, or pharmaceutically acceptable salts thereof.

[0299] Table 1.

[0300]

[0301]

[0302]

[0303]

[0304]

[0305]

[0306]

[0307]

[0308]

[0309]

[0310]

[0311] The compounds of the present invention can generally be prepared or isolated by synthetic and / or semi-synthetic methods known to those skilled in the art for similar compounds and by the methods described in detail in the examples herein. In some embodiments, the present invention provides intermediate compounds or salts thereof described in the examples.

[0312] 4. Usage, preparation and application:

[0313] Pharmaceutically acceptable compositions

[0314] According to another embodiment, the present invention provides a pharmaceutical 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 the compound in the composition of the present invention is effective to measurably inhibit USP1 or a variant or mutant thereof in a biological sample or patient. In certain embodiments, the amount of the compound in the composition of the present invention is effective to measurably inhibit USP1 or a variant or mutant thereof in a biological sample or 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.

[0315] As used herein, the term "patient" or "subject" means an animal, preferably a mammal, and most preferably a human.

[0316] 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 formulated therewith. 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 silicon dioxide, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol and lanolin).

[0317] "Pharmaceutically acceptable derivative" means any non-toxic salt, ester, salt of an ester, or other derivative of a compound of the invention that, when administered to a recipient, is capable of providing, directly or indirectly, a compound of the invention or an inhibitory active metabolite or residue thereof.

[0318] As used herein, the term "inhibitory active metabolite or residue thereof" means that the metabolite or residue thereof is also an inhibitor of USP1 or a variant or mutant thereof.

[0319] The compositions of the present invention can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, orally, vaginally, or via an implanted reservoir. As used herein, the term "parenteral" includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally, or intravenously. The sterile injectable form of the compositions of the present invention can be an aqueous or oily suspension. These suspensions can be formulated using appropriate dispersants or wetting agents and suspending agents according to techniques known in the art. Sterile injectable formulations can also be sterile injectable solutions or suspensions in non-toxic parenteral acceptable diluents or solvents, for example, in the form of solutions in 1,3-butanediol. Acceptable vehicles and solvents that can be used are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally used as solvents or suspending media.

[0320] For this purpose, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. For example, fatty acids such as oleic acid and their glyceride derivatives can be used to prepare injectables, as can natural pharmaceutically acceptable oils (such as olive oil or castor oil, especially their polyoxyethylated forms). These oil solutions or suspensions can also contain long-chain alcohol diluents or dispersants, such as carboxymethyl cellulose or similar dispersants commonly used in the preparation of pharmaceutically acceptable dosage forms (including emulsions and suspensions). For the purpose of formulation, other commonly used surfactants (such as Tween, Span) and other emulsifiers or bioavailability enhancers commonly used in the manufacture of pharmaceutically acceptable solids, liquids or other dosage forms can also be used.

[0321] The pharmaceutically acceptable compositions 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. In the case of tablets for oral use, conventional 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 dry corn starch. When an aqueous suspension is required for oral use, the active ingredient is combined with an emulsifier and a suspending agent. If desired, certain sweeteners, flavorings, or coloring agents may also be added.

[0322] Alternatively, the pharmaceutically acceptable compositions of the present invention can be administered in the form of suppositories for rectal administration. These suppositories can be prepared by mixing the medicament with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and will therefore melt in the rectum to release the drug. Such materials include cocoa butter, beeswax, and polyethylene glycol.

[0323] The pharmaceutically acceptable compositions of the present invention can also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, skin, or lower intestinal tract. Suitable topical formulations for each of these areas or organs are readily prepared.

[0324] Topical application to the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation.Topical transdermal patches may also be used.

[0325] For topical application, the pharmaceutically acceptable compositions provided can be formulated in a suitable ointment containing an active ingredient suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of the present invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsifying wax, and water. Alternatively, the pharmaceutically acceptable compositions provided can be formulated in a suitable lotion or cream form containing an active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and water.

[0326] For ophthalmic use, provided pharmaceutically acceptable compositions can be formulated as micronized suspensions or preferably as solutions in pH-adjusted isotonic sterile saline with or without a preservative such as benzalkonium chloride. Alternatively, for ophthalmic use, the pharmaceutically acceptable compositions can be formulated in an ointment such as petrolatum.

[0327] The pharmaceutically acceptable compositions of the present invention may also be administered by nasal spray or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation and may be prepared as solutions in normal saline using benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents.

[0328] Most preferably, pharmaceutically acceptable compositions of the present invention are formulated for oral administration. Such formulations may be administered with or without food. In certain embodiments, pharmaceutically acceptable compositions of the present invention are not administered with food. In other embodiments, pharmaceutically acceptable compositions of the present invention are administered with food.

[0329] The amount of the compound of the invention that can be combined with the carrier materials to produce a composition in a single dosage form varies depending on the host being treated, the particular mode of administration. Preferably, provided compositions should be formulated so that a dosage of 0.01-100 mg / kg body weight / day of the inhibitor can be administered to a patient receiving these compositions.

[0330] It will 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, age, weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician, as well as the severity of the specific disease being treated. The amount of the compound of the invention in the composition will also depend on the specific compound in the composition.

[0331] Uses of compounds and pharmaceutically acceptable compositions

[0332] According to another embodiment, the present invention provides a method for treating a disease or condition associated with USP1 in a patient, the method comprising administering to the patient a compound of the present invention, or a pharmaceutically acceptable derivative thereof, or a pharmaceutical composition thereof. In some embodiments, the present invention provides the use of a compound of the present invention, or a pharmaceutically acceptable derivative thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for treating a disease or condition associated with USP1. In some embodiments, the present invention provides the use of a compound of the present invention, or a pharmaceutically acceptable derivative thereof, or a pharmaceutical composition thereof, for treating a disease or condition associated with USP1. In some embodiments, the disease or condition associated with USP1 is a proliferative disease, such as a cancer as described herein.

[0333] As used herein, the terms "treatment," "treat," and "treating" refer to reversing, alleviating, delaying the onset of, or inhibiting the progression of, a disease or condition as described herein, or one or more symptoms thereof. In some embodiments, treatment may be administered after the development of one or more symptoms. 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., based on a history of symptoms and / or based on genetic or other predisposing factors). Treatment may also be continued after symptoms have resolved, for example, to prevent or delay their recurrence.

[0334] As used herein, "proliferative disease" refers to a disease that occurs due to abnormal growth or extension of cell proliferation (Walker, Cambridge Dictionary of Biology, Cambridge University Press: Cambridge, UK, 1990). Proliferative diseases may be associated with the following factors: 1) pathological proliferation of normal quiescent cells; 2) pathological migration of cells from their normal location (e.g., metastasis of tumor cells); 3) pathological expression of proteolytic enzymes, such as matrix metalloproteinases (e.g., collagenases, gelatinases, and elastases); or 4) pathological angiogenesis such as in proliferative retinopathy and tumor metastasis. Exemplary proliferative diseases include cancer (i.e., "malignant tumors"), benign tumors, angiogenesis, inflammatory diseases, and autoimmune diseases.

[0335] cancer

[0336] Cancers or proliferative diseases or tumors treated using the compounds, methods, and uses described herein include, but are not limited to, hematologic cancers, lymphomas, myelomas, leukemias, neurological cancers, skin cancers, breast cancers, prostate cancers, colorectal cancers, lung cancers, head and neck cancers, gastrointestinal cancers, liver cancers, pancreatic cancers, genitourinary cancers, bone cancers, kidney cancers, and vascular cancers.

[0337] In some embodiments of the methods and uses described herein, the cancer is lung cancer, thyroid cancer, ovarian cancer, colorectal cancer, prostate cancer, pancreatic cancer, esophageal cancer, liver cancer, breast cancer, skin cancer, or mesothelioma. In some embodiments, the cancer is lung cancer, thyroid cancer, ovarian cancer, colorectal cancer, prostate cancer, pancreatic cancer, esophageal cancer, liver cancer, breast cancer, skin cancer, or mesothelioma, sarcoma, or epithelioid hemangioendothelioma (EHE). In some embodiments, the cancer is mesothelioma, such as malignant mesothelioma. In some embodiments, the cancer is EHE.

[0338] In some embodiments, cancer includes, but is not limited to, leukemias (e.g., acute leukemias, acute lymphocytic leukemias, acute myeloid leukemias, acute myeloblastic leukemias, acute promyelocytic leukemias, acute myelomonocytic leukemias, acute monocytic leukemias, acute erythroleukemias, chronic leukemias, chronic myeloid leukemias, chronic lymphocytic leukemias), polycythemia vera, lymphomas (e.g., Hodgkin's disease or non-Hodgkin's disease), Waldenstrom's macroglobulinemia, multiple myeloma, heavy chain diseases, and solid tumors such as sarcomas and carcinomas (e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's sarcoma, flat Lymphosarcoma, rhabdomyosarcoma, colon cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, uterine cancer, testicular cancer, lung cancer, small cell lung cancer, bladder cancer, epithelial carcinoma, glioma, astrocytoma, glioblastoma multiforme (GBM, also known as glioblastoma), medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma, melanoma, neuroblastoma, and retinoblastoma).

[0339] In some embodiments, the cancer is a glioma, astrocytoma, glioblastoma multiforme (GBM, also known as glioblastoma), medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma, melanoma, neuroblastoma, or retinoblastoma.

[0340] In some embodiments, the cancer is an acoustic neuroma, an astrocytoma (e.g., grade I-pilocytic astrocytoma, grade II-low-grade astrocytoma, grade III-anaplastic astrocytoma, or grade IV-glioblastoma (GBM)), a chordoma, a CNS lymphoma, a craniopharyngioma, a brainstem glioma, an ependymoma, a mixed glioma, an optic nerve glioma, a subependymoma, a medulloblastoma, a meningioma, a metastatic brain tumor, an oligodendroglioma, a pituitary tumor, a primitive neuroectodermal (PNET) tumor, or a schwannoma. In some embodiments, the cancer is a type that is more common in children than in adults, such as a brainstem glioma, a craniopharyngioma, an ependymoma, a juvenile pilocytic astrocytoma (JPA), a medulloblastoma, an optic nerve glioma, a pineal tumor, a primitive neuroectodermal tumor (PNET), or a rhabdoid tumor. In some embodiments, the patient is an adult. In some embodiments, the patient is a child or a pediatric patient.

[0341] In another embodiment, cancer includes, but is not limited to, mesothelioma, hepatobiliary cancer (liver and bile duct), bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, ovarian cancer, colon cancer, rectal cancer, anal cancer, stomach cancer, gastrointestinal cancer (stomach cancer, colorectal cancer, and duodenal cancer), uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, kidney cancer, thyroid carcinoma, and thyroid carcinoma. adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, testicular cancer, chronic or acute leukemia, chronic myeloid leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal cell carcinoma, renal pelvis cancer, non-Hodgkin lymphoma, spinal axis tumors, brain stem glioma, pituitary adenoma, adrenocortical carcinoma, gallbladder cancer, multiple myeloma, bile duct cancer, fibrosarcoma, neuroblastoma, retinoblastoma, or a combination of one or more of the foregoing cancers.

[0342] In some embodiments, the cancer is selected from hepatocellular carcinoma, ovarian cancer, epithelial ovarian cancer, or fallopian tube cancer; papillary serous cystadenocarcinoma or uterine papillary serous carcinoma (UPSC); prostate cancer; testicular cancer; gallbladder cancer; hepatobiliary duct cancer; synovial sarcoma of soft tissue and bone; rhabdomyosarcoma; osteosarcoma; chondrosarcoma; Ewing sarcoma; anaplastic thyroid carcinoma; adrenocortical adenoma; pancreatic cancer; pancreatic ductal carcinoma or pancreatic adenocarcinoma; gastrointestinal tract / gastric (GIST) cancer; lymphoma; squamous cell carcinoma of the head and neck (SCCHN); salivary gland cancer; glioma or brain cancer; neurofibromatosis-1-associated malignant peripheral nerve sheath tumor (MPNST); Waldenstrom's macroglobulinemia; or medulloblastoma.

[0343] In some embodiments, the cancer is selected from hepatocellular carcinoma (HCC), hepatoblastoma, colon cancer, rectal cancer, ovarian cancer, epithelial ovarian cancer, fallopian tube cancer, papillary serous cystadenocarcinoma, uterine papillary serous carcinoma (UPSC), hepatobiliary carcinoma, synovial sarcoma of soft tissue and bone, rhabdomyosarcoma, osteosarcoma, anaplastic thyroid carcinoma, adrenocortical adenoma, pancreatic cancer, pancreatic ductal carcinoma, pancreatic adenocarcinoma, glioma, neurofibromatosis-1 associated malignant peripheral nerve sheath tumor (MPNST), Waldenstrom's macroglobulinemia, or medulloblastoma.

[0344] In some embodiments, the cancer is a solid tumor, such as a sarcoma, carcinoma, or lymphoma. Solid tumors generally contain abnormal tissue masses and generally do not include cysts or fluid areas. In some embodiments, the cancer is selected from renal cell carcinoma or kidney cancer; hepatocellular carcinoma (HCC) or hepatoblastoma, or liver cancer; melanoma; breast cancer; colorectal tumors, or colorectal cancer; colon cancer; rectal cancer; anal cancer; lung cancer, such as non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC); ovarian cancer, ovarian epithelial cancer, ovarian tumors, or fallopian tube cancer; papillary serous cystadenocarcinoma or uterine papillary serous carcinoma (UPSC); prostate cancer; testicular cancer; Gallbladder cancer; hepatobiliary cancer; synovial sarcoma of soft tissue and bone; rhabdomyosarcoma; osteosarcoma; chondrosarcoma; Ewing sarcoma; anaplastic thyroid cancer; adrenocortical carcinoma; pancreatic cancer; pancreatic ductal carcinoma or pancreatic adenocarcinoma; gastrointestinal tract / gastric (GIST) cancer; lymphoma; squamous cell carcinoma of the head and neck (SCCHN); salivary gland cancer; glioma or brain cancer; neurofibromatosis-1-associated malignant peripheral nerve sheath tumor (MPNST); Waldenstrom's macroglobulinemia; or medulloblastoma.

[0345] In some embodiments, the cancer is selected from renal cell carcinoma, hepatocellular carcinoma (HCC), hepatoblastoma, colorectal tumor, colorectal cancer, colon cancer, rectal cancer, anal cancer, ovarian cancer, ovarian epithelial cancer, ovarian tumor, fallopian tube cancer, papillary serous cystadenocarcinoma, uterine papillary serous carcinoma (UPSC), hepatobiliary carcinoma, synovial sarcoma of soft tissue and bone, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, anaplastic thyroid cancer, adrenocortical carcinoma, pancreatic cancer, pancreatic ductal carcinoma, pancreatic adenocarcinoma, glioma, brain cancer, neurofibromatosis-1 associated malignant peripheral nerve sheath tumor (MPNST), Waldenstrom's macroglobulinemia, or medulloblastoma.

[0346] In some embodiments, the cancer is selected from hepatocellular carcinoma (HCC), hepatoblastoma, colon cancer, rectal cancer, ovarian cancer, ovarian epithelial cancer, ovarian tumors, fallopian tube cancer, papillary serous cystadenocarcinoma, uterine papillary serous carcinoma (UPSC), hepatobiliary carcinoma, synovial sarcoma of soft tissue and bone, rhabdomyosarcoma, osteosarcoma, anaplastic thyroid cancer, adrenocortical carcinoma, pancreatic cancer, pancreatic ductal carcinoma, pancreatic adenocarcinoma, glioma, neurofibromatosis-1 associated malignant peripheral nerve sheath tumor (MPNST), Waldenstrom's macroglobulinemia, or medulloblastoma.

[0347] In some embodiments, the cancer is hepatocellular carcinoma (HCC). In some embodiments, the cancer is hepatoblastoma. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is rectal cancer. In some embodiments, the cancer is ovarian cancer or an ovarian tumor. In some embodiments, the cancer is epithelial ovarian cancer. In some embodiments, the cancer is fallopian tube cancer. In some embodiments, the cancer is papillary serous cystadenocarcinoma. In some embodiments, the cancer is uterine papillary serous carcinoma (UPSC). In some embodiments, the cancer is hepatobiliary duct cancer. In some embodiments, the cancer is synovial sarcoma of soft tissue and bone. In some embodiments, the cancer is rhabdomyosarcoma. In some embodiments, the cancer is osteosarcoma. In some embodiments, the cancer is anaplastic thyroid carcinoma. In some embodiments, the cancer is adrenocortical carcinoma. In some embodiments, the cancer is pancreatic cancer or pancreatic ductal carcinoma. In some embodiments, the cancer is pancreatic adenocarcinoma. In some embodiments, the cancer is glioma. In some embodiments, the cancer is malignant peripheral nerve sheath tumor (MPNST). In some embodiments, the cancer is neurofibromatosis-1-associated MPNST. In some embodiments, the cancer is Waldenstrom's macroglobulinemia. In some embodiments, the cancer is medulloblastoma.

[0348] In some embodiments, the cancer is a virus-associated cancer, including human immunodeficiency virus (HIV)-associated solid tumors, human papillomavirus (HPV)-16-positive incurable solid tumors, and adult T-cell leukemia, which is caused by human T-cell leukemia virus type 1 (HTLV-I) and is a highly aggressive form of CD4+ T-cell leukemia characterized by clonal integration of HTLV-I in leukemia cells (see https: / / clinicaltrials.gov / ct2 / show / study / NCT02631746); as well as virus-associated tumors in gastric cancer, nasopharyngeal cancer, cervical cancer, vaginal cancer, vulvar cancer, head and neck squamous cell carcinoma, and Merkel cell carcinoma. (See https: / / clinicaltrials.gov / ct2 / show / study / NCT02488759; also see https: / / clinicaltrials.gov / ct2 / show / study / NCT0240886 and https: / / clinicaltrials.gov / ct2 / show / NCT02426892)

[0349] In some embodiments, the cancer is melanoma. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is small cell lung cancer (SCLC). In some embodiments, the cancer is non-small cell lung cancer (NSCLC).

[0350] The compounds and compositions according to the methods of the present invention can be administered in any amount and by any route of administration that is effective in treating or reducing the severity of a cancer or tumor. The exact amount required varies from subject to subject, depending on the species, age, and general condition of the subject, the severity of the disease or condition, the specific agent, its mode of administration, and the like. The compounds of the present invention are preferably formulated in dosage unit form for ease of administration and dosage uniformity. As used herein, the expression "dosage unit form" refers to a physically discrete unit of a drug suitable for the patient to be treated. However, it should be understood that the total daily dosage of the compounds and compositions of the present invention is determined by the attending physician within the scope of sound medical judgment. The specific effective dosage level for any particular patient or organism depends on a variety of factors, including: the condition being treated and the severity of the condition; the activity of the specific compound being used; the specific composition being used; the patient's age, weight, general health, sex, and diet; the time of administration, route of administration, and excretion rate of the specific compound being used; the duration of treatment; drugs used in combination with or concurrently with the specific compound being used, and similar factors well known in the medical field. As used herein, the term "patient" or "subject" means an animal, preferably a mammal, and most preferably a human.

[0351] The pharmaceutically acceptable compositions of the present invention may be administered to humans and other animals orally, rectally, parenterally, intracisternal, intravaginal, intraperitoneally, topically (e.g., by powder, ointment, or drops), buccally, as an oral or nasal spray, etc., depending on the severity of the disease or condition being treated. In certain embodiments, the compounds of the present invention may be administered orally or parenterally at a dosage level 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, once or more daily to achieve the desired therapeutic effect.

[0352] 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, the liquid dosage form may also contain inert diluents commonly used in the art, such as water or other solvents, solubilizers, and emulsifiers, such as ethanol, 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), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and sorbitan fatty acid esters and mixtures thereof. In addition to inert diluents, oral compositions may also include adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavorings, and fragrances.

[0353] Injectable preparations can be prepared according to known techniques using suitable dispersants or wetting agents and suspending agents, such as sterile injectable aqueous or oily suspensions. Sterile injectable preparations can also be sterile injectable solutions, suspensions or emulsions in non-toxic parenteral acceptable diluents or solvents, such as in the form of solutions in 1,3-butanediol. Acceptable vehicles and solvents that can be used are water, Ringer's solution, USP and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally used as solvents or suspension media. For this purpose, any gentle fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids, such as oleic acid, are used when preparing injectables.

[0354] The 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 that can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0355] In order to prolong the effect of the compound of the present invention, it is often necessary to slow down the absorption of the compound from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous materials with poor water solubility. The absorption rate of the compound then depends on its dissolution rate, which in turn can depend on the crystal size and crystalline form. Alternatively, delayed absorption of the compound form administered parenterally is achieved by dissolving or suspending the compound in an oil vehicle. Injectable accumulation forms are prepared by forming a microcapsule matrix of the compound in a biodegradable polymer (e.g., polylactide-polyglycolide). Depending on the ratio of the compound to the polymer and the properties of the specific polymer used, the release rate of the compound can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Accumulated injectable formulations are also prepared by coating the compound in liposomes or microemulsions compatible with body tissues.

[0356] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

[0357] 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, polyvinyl pyrrolidone, sucrose and acacia, c) humectants such as glycerol, d) disintegrants such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glyceryl monostearate, h) absorbents such as kaolin and bentonite, 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 form may also include a buffer.

[0358] Also can adopt similar type solid composition as the filler in soft-filled gelatin capsule and the hard-filled gelatin capsule using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol.The tablet, dragee, capsule, pill and granule of solid dosage form can use coating and shell, for example other coating well-known in enteric coating and pharmaceutical preparation field is prepared.They can optionally contain opacifier, and can also have the composition that makes it optionally only or preferentially release one or more active components in a certain part of intestinal tract in a delayed manner.The example of operable embedded composition comprises polymeric substance and wax.Also can adopt similar type solid composition as the filler in soft-filled gelatin capsule and the hard-filled gelatin capsule using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol.

[0359] The active compound can also be in the form of microencapsulation with one or more excipients as mentioned above. Tablets, dragees, capsules, pills and granules of solid dosage forms can be prepared with coatings and shells, such as enteric coatings, release-controlled coatings and other coatings known in the field of pharmaceutical formulations. In such solid dosage forms, the active compound can be mixed with at least one inert diluent (such as sucrose, lactose or starch). During normal practice, such dosage forms can also include other substances other than the inert diluent, 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 form can also include a buffer. It can optionally contain an emulsifier and can also have a composition that allows it to optionally only or preferentially release one or more active ingredients in a delayed manner in a certain part of the intestinal tract. Examples of usable embedding compositions include polymeric substances and waxes.

[0360] The dosage form for topical or transdermal administration of the compound of the present invention includes ointment, paste, cream, lotion, gel, powder, solution, spray, inhalant or patch. Under aseptic conditions, active ingredient and pharmaceutically acceptable carrier and any required preservative or buffer are blended as required. Ophthalmic formulations, ear drops and eye drops are also contemplated to be within the scope of the present invention. In addition, the present invention contemplates the use of transdermal patches, which have the added advantage of enabling the compound to be controllably delivered to health. The dosage form can be manufactured by dissolving the compound or being assigned in an appropriate medium. Absorption enhancers can also be used to increase the flow of the compound through the skin. Rate control can be achieved by providing a rate-controlled membrane or by dispersing the compound in a polymer matrix or a gel.

[0361] Co-administered with one or more other therapeutic agents

[0362] Depending on the specific condition or disease to be treated, additional therapeutic agents that are normally administered to treat that condition may also be present in the compositions of the invention. As used herein, additional therapeutic agents that are normally administered to treat a particular disease or condition are known as "appropriate for the disease or condition being treated."

[0363] In some embodiments, the present invention provides a method of treating a disclosed disease or condition, comprising administering to a patient in need thereof an effective amount of a compound disclosed herein or a pharmaceutically acceptable salt thereof, and simultaneously or sequentially administering an effective amount of one or more additional therapeutic agents, such as those described herein. In some embodiments, the method comprises co-administering one additional therapeutic agent. In some embodiments, the method comprises co-administering two additional therapeutic agents. In some embodiments, the combination of the disclosed compound and the additional one or more therapeutic agents acts synergistically.

[0364] The compounds of the present invention may also be used in combination with known treatments, such as administration of hormones or radiation. In certain embodiments, provided compounds are used as radiosensitizers, particularly for treating tumors that exhibit poor sensitivity to radiation therapy.

[0365] The compounds of the present invention can be administered alone or in combination with one or more other therapeutic compounds, and possible combination therapies may provide the administration of the compounds of the present invention and one or more other therapeutic compounds in a fixed combination form or staggered or independently of one another, or a fixed combination and one or more other therapeutic compounds may be administered in combination. The compounds of the present invention may be administered in addition or in addition, particularly in combination with chemotherapy, radiotherapy, immunotherapy, phototherapy, surgical intervention, or a combination of these for tumor therapy. In the case of other treatment strategies, long-term therapy and adjuvant therapy are also possible, as described above. Other possible treatments are therapies or even chemopreventive therapies (e.g., for patients at risk) that maintain the patient's status after tumor regression.

[0366] One or more other therapeutic agents can be used separately with the compound of the present invention or composition, as a part for multiple dose regimen.Alternatively, one or more other therapeutic agents can be a part for a single dosage form, mixed in a single composition together with the compound of the present invention.If used as multiple dose regimen, one or more other therapeutic agents and the compound of the present invention or composition can be used simultaneously, sequentially or in a period of time spaced apart from each other (for example, in 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,18,20,21,22,23 or 24 hours spaced apart from each other).In certain embodiments, one or more other therapeutic agents and the compound of the present invention or composition are used as multiple dose regimen at intervals greater than 24 hours.

[0367] As used herein, the terms "combination," "combined," and related terms refer to the simultaneous or sequential administration of therapeutic agents according to the present invention. For example, a compound of the present invention can be administered in separate unit dosage forms or in a single unit dosage form together with one or more other therapeutic agents, either simultaneously or sequentially. Thus, the present invention provides a single unit dosage form comprising a compound of the present invention, one or more other therapeutic agents, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

[0368] The amount of the compound of the invention and one or more other therapeutic agents (in those compositions comprising additional therapeutic agents as described above) that can be combined with the carrier materials to produce a single dosage form will vary depending on the host being treated and the specific mode of administration. Preferably, the compositions of the invention should be formulated so that a dosage of the compound of the invention between 0.01-100 mg / kg body weight / day can be administered.

[0369] In those compositions comprising one or more additional therapeutic agents, the one or more additional therapeutic agents and the compounds of the present invention may act synergistically. Thus, the amount of the one or more additional therapeutic agents in such compositions may be less than the amount required in a monotherapy utilizing only that therapeutic agent. In such compositions, the one or more additional therapeutic agents may be administered at a dosage of between 0.01–1,000 μg / kg body weight / day.

[0370] The amount of one or more other therapeutic agents present in the composition of the present invention may not exceed the amount normally administered in a composition comprising the therapeutic agent as the sole active agent. Preferably, the amount of one or more other therapeutic agents in the composition of the present disclosure will be in the range of about 50% to 100% of the amount normally present in a composition comprising the drug as the sole therapeutic active agent. In certain embodiments, one or more other therapeutic agents are administered at a dosage of about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90% or about 95% of the normal administration amount of the agent. As used herein, the phrase "normal administration" means the amount of an FDA-approved therapeutic agent approved for administration according to the FDA label insert.

[0371] The compounds of the present invention or their pharmaceutical compositions can also be incorporated into compositions for coating implantable medical devices, such as prostheses, artificial valves, artificial blood vessels, stents, and catheters. Vascular stents, for example, 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 forming clots or platelet activation. These undesirable effects can be prevented or alleviated by pre-coating the device with a pharmaceutically acceptable composition comprising a kinase inhibitor. An implantable device coated with a compound of the present invention is another embodiment of the present invention.

[0372] Exemplary Additional Therapeutic Agents

[0373] In some embodiments, the one or more additional therapeutic agents are poly ADP ribose polymerase (PARP) inhibitors. In some embodiments, the PARP inhibitor is selected from olaparib ( AstraZeneca; Lucaparib ( Clovis Oncology; Niraparib ( Tesaro); talazoparib (MDV3800 / BMN673 / LT00673, Medivation / Pfizer / Biomarin); veliparib (ABT-888, AbbVie); and BGB-290 (BeiGene, Inc.).

[0374] In some embodiments, the one or more other therapeutic agents are histone deacetylase (HDAC) inhibitors. In some embodiments, the HDAC inhibitor is selected from vorinostat ( Merck); Romidepsin ( Celgene); Panobinostat ( Novartis; Belinostat ( Spectrum Pharmaceuticals); entinostat (SNDX-275, Syndax Pharmaceuticals) (NCT00866333); and chidamide ( HBI-8000, Chipscreen Biosciences, China).

[0375] In some embodiments, the one or more additional therapeutic agents are CDK inhibitors, such as CDK4 / CDK6 inhibitors. In some embodiments, the CDK 4 / 6 inhibitor is selected from palbociclib ( Pfizer; Ribociclib ( Novartis); abemaciclib (Ly2835219, Eli Lilly); and trilaciclib (G1T28, G1 Therapeutics).

[0376] In some embodiments, the one or more additional therapeutic agents are phosphatidylinositol 3-kinase (PI3K) inhibitors. In some embodiments, the PI3K inhibitor is selected from idelalisib ( Gilead), apellisib (BYL719, Novartis), taselisib (GDC-0032, Genentech / Roche); pictilisib (GDC-0941, Genentech / Roche); copanlisib (BAY806946, Bayer); duvelisib (formerly IPI-145, Infinity Pharmaceuticals); PQR309 (Piqur Therapeutics, Switzerland); and TGR1202 (formerly RP5230, TG Therapeutics).

[0377] In some embodiments, the one or more additional therapeutic agents are platinum-based therapeutic agents, also known as platinum. Platinums cause cross-linking of DNA, thereby inhibiting DNA repair and / or DNA synthesis, particularly in rapidly multiplying cells, such as cancer cells.

[0378] In some embodiments, the platinum-based therapeutic agent is selected from cisplatin ( Bristol-Myers Squibb); Carboplatin ( Bristol-Myers Squibb; Teva, Pfizer); oxaliplatin ( Sanofi-Aventis); Nedaplatin ( Shionogi), picoplatin (Poniard Pharmaceuticals); and satraplatin (JM-216, Agennex).

[0379] In some embodiments, the one or more additional therapeutic agents are taxane compounds that cause disruption of microtubules necessary for cell division. In some embodiments, the taxane compound is selected from paclitaxel ( Bristol-MyersSquibb), docetaxel ( Sanofi-Aventis; Sun Pharmaceutical), albumin-bound paclitaxel ( Abraxis / Celgene), Cabazitaxel ( Sanofi-Aventis) and SID530 (SK Chemicals, Co.) (NCT00931008).

[0380] In some embodiments, the one or more additional therapeutic agents are nucleoside inhibitors, or therapeutic agents that interfere with normal DNA synthesis, protein synthesis, cell replication, or would otherwise inhibit rapidly proliferating cells.

[0381] In some embodiments, the nucleoside inhibitor is selected from trabectedin (a guanidinating agent, JanssenOncology), nitrogen mustard (alkylating agent, Aktelion Pharmaceuticals); vincristine ( Eli Lilly; Teva Pharmaceuticals; Talon Therapeutics); Temozolomide (a prodrug of the alkylating agent 5-(3-methyltriazene-1-yl)-imidazole-4-carboxamide (MTIC), Merck); cytarabine injection (ara-C, an antimetabolite cytidine analog, Pfizer); lomustine (alkylating agent, Bristol-Myers Squibb; NextSource Biotechnology); azacitidine (a pyrimidine nucleoside analog of cytidine, Celgene); omacetaxine mepesuccinate (cephalotaxine ester) (protein synthesis inhibitor, Teva Pharmaceuticals); asparaginase Erwinia chrysanthemi (enzyme for asparagine depletion, Lundbeck; EUSA Pharma); Eribulin mesylate (microtubule inhibitor, tubulin-based antimitotic agent, Eisai); Cabazitaxel (microtubule inhibitor, tubulin-based antimitotic agent, Sanofi-Aventis; capacetrine (thymidylate synthase inhibitor, Genentech); bendamustine (a bifunctional nitrogen mustard derivative believed to form interstrand DNA crosslinks, Cephalon / Teva); Ixabepilone (a semisynthetic analog of epothilone B, a microtubule inhibitor, and a tubulin-based antimitotic agent, Bristol-Myers Squibb); Nelarabine (a prodrug of a deoxyguanosine analog, a nucleoside metabolism inhibitor, Novartis); clorafabine (a prodrug of a ribonucleotide reductase inhibitor, a competitive inhibitor of deoxycytidine, Sanofi-Aventis); and trifluridine and tipiracil (thymidine-based nucleoside analogs and thymidine phosphorylase inhibitors, Taiho Oncology).

[0382] In some embodiments, one or more other therapeutic agents are kinase inhibitors or VEGF-R antagonists. In some embodiments, one or more other therapeutic agents are MEK inhibitors. As used herein, "MEK inhibitor" refers to any inhibitor or blocker or antagonist that binds to and / or inhibits mitogen-activated protein kinases MEK1 and / or MEK2. In some embodiments, the MEK inhibitor is selected from those described in Cheng et al., "Current Development Status of MEK Inhibitors," Molecules 2017, 22, 1551, the contents of which are incorporated herein by reference in their entirety. In certain embodiments, the MEK inhibitor is selected from binimetinib (MEK162, ARRY-438162, ARRAY BIOPHARMA INC.), cobimetinib ( Exelexis / Genentech / Roche), refametinib (BAY 86-9766, RDEA119; Bayer AG), selumetinib (AZD6244, ARRY-142886; ASTRAZENECA), trametinib ( In some embodiments, the second anticancer agent is mirdametinib (Novatinib), mirdametinib (PD-0325901, Spring Works Therapeutics), pimasertib (AS703026, MSC1936369B, Merck KGaA), or any pharmaceutically acceptable salts and / or solvates thereof. In certain embodiments, the second anticancer agent is binimetinib, cobimetinib, selumetinib, trametinib, mirdametinib, pimasertib, or any pharmaceutically acceptable salts and / or solvates thereof. Other examples of MEK inhibitors for use as additional therapeutic agents in the methods and uses described herein include, but are not limited to, E6201 (Eisai Co Ltd. / Strategia Theraputics), GDC-0623 (RG 7421, Genentech, Inc.), CH5126766 (RO5126766, Chugai 232 Pharmaceutical Co., Roche), HL-085 (Shanghai Kechow Pharma, Inc.), SHR7390 (HENGRUI MEDICINE), TQ-B3234 (CHIATAI TIANQING), CS-3006 (CSTONE Pharmaceuticals), FCN-159 (Fosun Pharmaceuticals), VS-6766 (Verastem Oncology), and IMM-1-104 (Immuneering Corp.). Other examples of MEK inhibitors for use as a second anticancer agent in the methods and uses described herein include, but are not limited to, those described in WO2005 / 121142, WO2014 / 169843, WO2016 / 035008, WO2016 / 168704, WO2020 / 125747, WO2021 / 142144, WO2021 / 142345, WO2021 / 149776, the contents of each of which are incorporated herein by reference in their entirety.

[0383] In some embodiments, one or more other therapeutic agents are EGFR inhibitors. As used herein, "EGFR inhibitor" refers to any inhibitor or blocker or antagonist that binds to and / or inhibits epidermal growth factor receptor (EGFR). In some embodiments, the EGFR inhibitor is selected from Ayati et al., "A review on progression of epidermal growth factor receptor (EGFR) inhibitors as an efficient approach in cancer targeted therapy," Bioorganic Chemistry 2020, 99: Those described in 103811, the contents of which are incorporated herein by reference in their entirety. In some embodiments, the EGFR inhibitor is selected from cetuximab, nacituzumab, panitumumab, zalutumumab, nimotuzumab and matuzumab. In some embodiments, the EGFR inhibitor is cetuximab. In some embodiments, the EGFR inhibitor is nacituzumab. In some embodiments, the EGFR inhibitor is panitumumab. In some embodiments, the EGFR inhibitor is zalutumumab. In some embodiments, the EGFR inhibitor is nimotuzumab. In some embodiments, the EGFR inhibitor is matuzumab.

[0384] In some embodiments, the EGFR inhibitor is selected from osimertinib, gefitinib, erlotinib, lapatinib, neratinib, vandetanib, afatinib, brigatinib, dacomitinib, and icotinib. In some embodiments, the EGFR inhibitor is osimertinib. In some embodiments, the EGFR inhibitor is gefitinib. In some embodiments, the EGFR inhibitor is erlotinib. In some embodiments, the EGFR inhibitor is lapatinib. In some embodiments, the EGFR inhibitor is neratinib. In some embodiments, the EGFR inhibitor is vandetanib. In some embodiments, the EGFR inhibitor is afatinib. In some embodiments, the EGFR inhibitor is brigatinib. In some embodiments, the EGFR inhibitor is dacomitinib. In some embodiments, the EGFR inhibitor is icotinib.

[0385] In some embodiments, the EGFR inhibitor is a "first-generation EGFR tyrosine kinase inhibitor" (first-generation TKI). First-generation TKI refers to reversible EGFR inhibitors, such as gefitinib and erlotinib, which are effective first-line treatments for NSCLC harboring EGFR activating mutations (such as exon 19 deletions and exon 21 L858R mutations).

[0386] In some embodiments, the EGFR inhibitor is a "second-generation EGFR tyrosine kinase inhibitor" (second-generation TKI). Second-generation TKI refers to covalent irreversible EGFR inhibitors, such as afatinib and dacomitinib, which are effective first-line treatments for NSCLC harboring EGFR activating mutations (such as exon 19 deletions and exon 21 L858R mutations).

[0387] In some embodiments, the EGFR inhibitor is a "3rd generation EGFR tyrosine kinase inhibitor" (3rd generation TKI). 3rd generation TKI refers to covalent irreversible EGFR inhibitors, such as osimertinib and lazertinib, which are selective for EGFR activating mutations (such as exon 19 and exon 21 L858R deletions) alone or in combination with the T790M mutation and have lower inhibitory activity against wild-type EGFR.

[0388] In some embodiments, the one or more additional therapeutic agents are selected from approved VEGF inhibitors and kinase inhibitors that can be used in the present invention, including: bevacizumab ( Genentech / Roche), an anti-VEGF monoclonal antibody; ramucirumab ( Eli Lilly), an anti-VEGFR-2 antibody; and ziv-aflibercept, also known as VEGFTrap ( Regeneron / Sanofi). VEGFR inhibitors, such as regorafenib ( Bayer); vandetanib( AstraZeneca; Axitinib ( Pfizer; and lenvatinib ( Eisai); Raf inhibitors, such as sorafenib ( Bayer AG and Onyx); dabrafenib ( Novartis); and vemurafenib ( Genentech / Roche); MEK inhibitors such as cobimetinib ( Exelexis / Genentech / Roche); trametinib ( Novartis); Bcr-Abl tyrosine kinase inhibitors such as imatinib ( Novartis; Nilotinib ( Novartis; dasatinib ( BristolMyersSquibb; bosutinib ( Pfizer); and ponatinib ( Ariad Pharmaceuticals); Her2 and EGFR inhibitors, such as gefitinib ( AstraZeneca; Erlotinib ( Genentech / Roche / Astellas); Lapatinib ( Novartis; afatinib ( Boehringer Ingelheim); osimertinib (targets activated EGFR, AstraZeneca); and brigatinib ( Ariad Pharmaceuticals); c-Met and VEGFR2 inhibitors, such as cabozantanib ( Exelexis); and multikinase inhibitors, such as sunitinib ( Pfizer; Pazopanib ( Novartis); ALK inhibitors, such as crizotinib ( Pfizer; Ceritinib ( Novartis); and alectinib ( Genentech / Roche); Bruton's tyrosine kinase inhibitors, such as ibrutinib ( Pharmacyclos / Janssen); and Flt3 receptor inhibitors, such as mistalin ( Novartis).

[0389] Other kinase inhibitors and VEGF-R antagonists under development and useful in the present invention include tivozanib (Aveo Pharmaceuticals); vatalanib (Bayer / Novartis); lucitanib (Clovis Oncology); dovitinib (TKI258, Novartis); chiauanib (Chipscreen Biosciences); CEP-11981 (Cephalon); linifanib (Abbott Laboratories); neratinib (HKI-272, Puma Biotechnology); radotinib ( IY5511, Il-Yang Pharmaceuticals, South Korea); Ruxolitinib ( Incyte Corporation); PTC299 (PTC Therapeutics); CP-547,632 (Pfizer); foretinib (Exelexis, GlaxoSmithKline); quizartinib (Daiichi Sankyo) and motesanib (Amgen / Takeda).

[0390] In some embodiments, the one or more additional therapeutic agents is an mTOR inhibitor, which inhibits cell proliferation, angiogenesis, and glucose uptake. In some embodiments, the mTOR inhibitor is everolimus ( Novartis); Temsirolimus ( Pfizer); and sirolimus ( Pfizer).

[0391] In some embodiments, the one or more additional therapeutic agents is a proteasome inhibitor. Approved proteasome inhibitors for use in the present invention include bortezomib ( Takeda); Carfilzomib ( Amgen); and ixazomib ( Takeda).

[0392] In some embodiments, the one or more additional therapeutic agents is a growth factor antagonist, such as an antagonist of platelet-derived growth factor (PDGF) or epidermal growth factor (EGF) or its receptor (EGFR). Approved PDGF antagonists that can be used in the present invention include olaratumab ( Eli Lilly). Approved EGFR antagonists that can be used in the present invention include cetuximab ( Eli Lilly; Nexituzumab ( Eli Lilly), panitumumab ( Amgen); and osimertinib (targeting activated EGFR, AstraZeneca).

[0393] In some embodiments, the one or more additional therapeutic agents are aromatase inhibitors. In some embodiments, the aromatase inhibitor is selected from exemestane ( Pfizer; anastazole ( AstraZeneca and letrozole ( Novartis).

[0394] In some embodiments, one or more additional therapeutic agents is an antagonist of the hedgehog pathway. Approved hedgehog pathway inhibitors that can be used in the present invention include sonidegib ( Sun Pharmaceuticals); and Vimodigibone ( Genentech), both used to treat basal cell carcinoma.

[0395] In some embodiments, the one or more other therapeutic agents is a folate inhibitor. Approved folate inhibitors for use in the present invention include pemetrexed ( Eli Lilly).

[0396] In some embodiments, the one or more additional therapeutic agents is a CC chemokine receptor 4 (CCR4) inhibitor. CCR4 inhibitors under investigation for use in the present invention include mogamulizumab ( Kyowa Hakko Kirin, Japan).

[0397] In some embodiments, the one or more additional therapeutic agents are isocitrate dehydrogenase (IDH) inhibitors. IDH inhibitors under investigation for use in the present invention include AG120 (Celgene; NCT02677922); AG221 (Celgene, NCT02677922; NCT02577406); BAY1436032 (Bayer, NCT02746081); IDH305 (Novartis, NCT02987010).

[0398] In some embodiments, the one or more other therapeutic agents are arginase inhibitors. Arginase inhibitors under investigation for use in the present invention include AEB1102 (PEGylated recombinant arginase, Aeglea Biotherapeutics), which is being studied in Phase 1 clinical trials for acute myeloid leukemia and myelodysplastic syndrome (NCT02732184) and solid tumors (NCT02561234); and CB-1158 (Calithera Biosciences).

[0399] In some embodiments, the one or more additional therapeutic agents is a glutaminase inhibitor. Glutaminase inhibitors under investigation for use in the present invention include CB-839 (Calithera Biosciences).

[0400] In some embodiments, the one or more additional therapeutic agents are antibodies that bind to tumor antigens (i.e., proteins expressed on the cell surface of tumor cells). Approved antibodies that bind to tumor antigens for use in the present invention include rituximab ( Genentech / BiogenIdec; ofatumumab (anti-CD20, GlaxoSmithKline); Obinutuzumab (anti-CD20, Genentech), ibrutinib (anti-CD20 and yttrium-90, Spectrum Pharmaceuticals); Daratumumab (anti-CD38, Janssen Biotech), denoximab (anti-glycolipid GD2, United Therapeutics; Trastuzumab (anti-HER2, Genentech; ado-trastuzumab emtansine (anti-HER2, fused to emtansine, Genentech); and pertuzumab (anti-HER2, Genentech); and brentuximab vedotin (anti-CD30 drug conjugate, Seattle Genetics).

[0401] In some embodiments, the one or more additional therapeutic agents are topoisomerase inhibitors. Approved topoisomerase inhibitors for use in the present invention include irinotecan ( Merrimack Pharmaceuticals; Topotecan ( GlaxoSmithKline). Topoisomerase inhibitors under investigation for use in the present invention include picroquinone ( CTI Biopharma).

[0402] In some embodiments, one or more additional therapeutic agents are inhibitors of anti-apoptotic proteins, such as BCL-2. Approved anti-apoptotic drugs that can be used in the present invention include venetoclax ( AbbVie / Genentech); and blinatumomab ( Other therapeutic agents targeting apoptotic proteins that have been clinically tested and can be used in the present invention include navitoclax (ABT-263, Abbott) and BCL-2 inhibitors (NCT02079740).

[0403] In some embodiments, the one or more additional therapeutic agents is an androgen receptor inhibitor. Approved androgen receptor inhibitors that can be used in the present invention include enzalutamide ( Astellas / Medivation); approved androgen synthesis inhibitors include abiraterone ( Centocor / Ortho); approved gonadotropin-releasing hormone (GnRH) receptor antagonists (degaralix, Ferring Pharmaceuticals).

[0404] In some embodiments, the one or more other therapeutic agents are selective estrogen receptor modulators (SERMs), which interfere with the synthesis or activity of estrogen. Approved SERMs for use in the present invention include raloxifene ( Eli Lilly).

[0405] In some embodiments, the one or more additional therapeutic agents is an inhibitor of bone resorption. Amgen) is an approved therapeutic agent that inhibits bone resorption. It is an antibody that binds to RANKL and prevents binding to its receptor RANK, which is present on the surface of osteoclasts, their precursors, and osteoclast-like giant cells and mediates bone pathology in solid tumors with bone metastases. Other approved therapeutic agents that inhibit bone resorption include bisphosphonates, such as zoledronic acid ( Novartis).

[0406] In some embodiments, one or more other therapeutic agents are inhibitors of the interaction between the two main p53 inhibitory proteins MDMX and MDM2. Inhibitors of p53 inhibitory proteins that are being studied and can be used in the present invention include ALRN-6924 (Aileron), which is a stapled peptide that binds to and destroys the interaction of MDMX and MDM2 with p53. ALRN-6924 is currently being evaluated in clinical trials for the treatment of AML, advanced myelodysplastic syndrome (MDS), and peripheral T-cell lymphoma (PTCL) (NCT02909972; NCT02264613).

[0407] In some embodiments, the one or more additional therapeutic agents are inhibitors of transforming growth factor-β (TGF-β or TGF-β). Investigational inhibitors of TGF-β proteins that may be used in the present invention include NIS793 (Novartis), an anti-TGF-β antibody that is being clinically tested for the treatment of various cancers, including breast cancer, lung cancer, hepatocellular carcinoma, colorectal cancer, pancreatic cancer, prostate cancer, and renal cancer (NCT 02947165). In some embodiments, the inhibitor of TGF-β protein is fresolimumab (GC1008; Sanofi-Genzyme), which is being studied for melanoma (NCT00923169); renal cell carcinoma (NCT00356460); and non-small cell lung cancer (NCT02581787). Additionally, in some embodiments, the additional therapeutic agent is a TGF-β trap, such as described in Connolly et al. (2012) Int'l J. Biological Sciences 8:964-978. One therapeutic compound currently in clinical trials for the treatment of solid tumors is M7824 (Merck KgaA - formerly known as MSB0011459X), a bispecific anti-PD-L1 / TGF-β trap compound (NCT02699515); and (NCT02517398). M7824 consists of a fully human IgG1 antibody against PD-L1 fused to the extracellular domain of human TGF-β receptor II, acting as a TGF-β "trap."

[0408] In some embodiments, the one or more additional therapeutic agents are selected from glembatumumab vedotin-monomethylreoxetine E (MMAE) (Celldex), an anti-glycoprotein NMB (gpNMB) antibody linked to cytotoxic MMAE (CR011). gpNMB is a protein overexpressed by various tumor types that is associated with the metastatic ability of cancer cells.

[0409] In some embodiments, the one or more additional therapeutic agents are anti-proliferative compounds. Such anti-proliferative compounds include, but are not limited to, 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; anti-tumor antimetabolites; platinum compounds; compounds that target / reduce the activity of protein or lipid kinases and other anti-angiogenic compounds; compounds that target, reduce or inhibit the activity of protein or lipid phosphatases; gonadorelin agonists; anti-androgens; methionine aminopeptidase inhibitors; matrix metalloproteinase inhibitors; bisphosphonates; biological response modifiers; anti-proliferative antibodies; heparanase inhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasome inhibitors; compounds for the treatment of hematological malignancies; compounds that target, reduce or inhibit the activity of Flt-3; Hsp90 inhibitors, such as those from Conforma Therapeutics' 17-AAG (17-allylaminogeldanamycin, NSC330507), 17-DMAG (17-dimethylaminoethylamino-17-demethoxy-geldanamycin, NSC707545), IPI-504, CNF1010, CNF2024, CNF1010; temozolomide Kinesin spindle protein inhibitors such as SB715992 or SB743921 from GlaxoSmithKline, or pentamidine / chlorpromazine from CombinatoRx; MEK inhibitors such as ARRY142886 from Array BioPharma, AZd6244 from AstraZeneca, PD181461 from Pfizer, and folinic acid.

[0410] The term "aromatase inhibitor" as used herein relates to a compound that inhibits estrogen production, for example the conversion of the substrates androstenedione and testosterone to estrone and estradiol, respectively. The term includes, but is not limited to, steroids, in particular atamestane, exemestane and formestane; and in particular non-steroidal, in particular aminoglutethimide, roglethimide, pyridoglutethimide, trilostane, testolactone, ketokonazole, vorozole, fadrozole, anastrozole and letrozole. Exemestane is marketed under the trade name Aromasin TM Formestane is sold under the brand name Lentaron TM Fadrozole is sold under the brand name Afema TM Anastrozole is sold under the brand name Arimidex TM Letrozole is sold under the brand name Femara TM or Femar TM Aminoglutethimide is sold under the brand name Orimeten TM The combinations of the invention comprising a chemotherapeutic agent that is an aromatase inhibitor are particularly useful in the treatment of hormone receptor positive tumors, such as breast tumors.

[0411] As used herein, the term "antiestrogen" refers to compounds that antagonize the effects of estrogens at the estrogen receptor level. The term includes, but is not limited to, tamoxifen, fulvestrant, raloxifene, and raloxifene hydrochloride. Tamoxifen is available under the trade name Nolvadex. TM Raloxifene hydrochloride is sold under the brand name Evista TM Fulvestrant is sold under the brand name Faslodex. TM Fulvestrant is available under the brand name Faslodex. TM Administration. Combinations of the invention comprising a chemotherapeutic agent that is an antiestrogen are particularly useful in the treatment of estrogen receptor positive tumors, such as breast tumors.

[0412] As used herein, the term "antiandrogen" refers to any substance capable of inhibiting the biological effects of androgens and includes, but is not limited to, bicalutamide (Casodex). TMAs used herein, the term "gonadotropin agonist" includes, but is not limited to, abarelix, goserelin, and goserelin acetate. Goserelin is available under the trade name Zoladex. TM Application.

[0413] As used herein, the term "topoisomerase I inhibitors" includes, but is not limited to, topotecan, gimatidecan, irinotecan, camptothecin and its analog 9-nitrocamptothecin and the macromolecular camptothecin conjugate PNU-166148. Irinotecan is available, for example, in the form in which it is sold (e.g., under the trademark Camptosar®). TM ) administration. Topotecan is sold under the brand name Hycamptin TM sell.

[0414] As used herein, the term "topoisomerase II inhibitors" include, but are not limited to, anthracyclines such as doxorubicin (including liposomal formulations such as Caelyx TM ), daunomycin, epirubicin, idarubicin, and nemorubicin, the anthraquinones mitoxantrone and losoxantrone, and the podophyllotoxins etoposide and teniposide. Etoposide is sold under the trade name Etopophos TM Teniposide is sold under the trade name VM 26-Bristol. Doxorubicin is sold under the trade name Acriblastin TM or Adriamycin TM Epirubicin is sold under the brand name Farmorubicin TM Idarubicin is sold under the brand name Zavedos TM Mitoxantrone is sold under the brand name Novantron TM sell.

[0415] The term "microtubule active agent" refers to microtubule stabilizing, microtubule destabilizing compounds and tubulin 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 available under the trade name Taxol TM Docetaxel is sold under the brand name Taxotere TM Vinblastine sulfate is sold under the trade name Vinblastin RP TM Sold. Vincristine sulfate is sold under the brand name Farmistin TM sell.

[0416] As used herein, the term "alkylating agent" includes, but is not limited to, cyclophosphamide, ifosfamide, melphalan, or nitrosourea (BCNU or Gliadel). Cyclophosphamide is sold under the trade name Cyclostin TM Ifosfamide is sold under the brand name Holoxan TM sell.

[0417] The term "histone deacetylase inhibitor" or "HDAC inhibitor" relates to compounds that inhibit histone deacetylase and have antiproliferative activity. This includes, but is not limited to, suberoylanilide hydroxamic acid (SAHA).

[0418] 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 folic acid antagonists such as pemetrexed. Capecitabine is sold under the trade name Xeloda TM Gemcitabine is sold under the brand name Gemzar. TM sell.

[0419] As used herein, the term "platinum compound" includes, but is not limited to, carboplatin, cis-platin, cisplatinum, and oxaliplatin. Carboplatin can be sold, for example, in the form in which it is sold (e.g., under the trademark Carboplat TM ) is administered. Oxaliplatin can be administered, for example, in the form in which it is sold (e.g. under the trademark Eloxatin TM ) application.

[0420] As used herein, the term "compounds that target / decrease the activity of protein or lipid kinases; or protein or lipid phosphatases; or other anti-angiogenic compounds" includes but is not limited to protein tyrosine kinase and / or serine and / or threonine kinase inhibitors or lipid kinase inhibitors, such as: a) compounds that target, decrease or inhibit the activity of platelet-derived growth factor receptor (PDGFR), such as compounds that target, decrease or inhibit the activity of PDGFR, in particular compounds that inhibit PDGF receptor, such as N-phenyl-2-pyrimidine-amine derivatives, such as imatinib, SU11, SU21, SU31, SU41, SU61, SU71, SU81, SU91, SU12, SU13, SU14, SU15, SU16, SU17, SU18, SU19, SU20, SU21, SU22, SU23, SU24, SU25, SU26, SU27, SU28, SU29, SU30, SU31, SU32, SU33, SU34, SU35, SU36, SU37, SU38, SU39, SU39, SU40, SU31, SU32, SU33, SU34, SU35, SU36, SU37, SU38, SU39, SU41, SU39, SU42, SU39, SU31, SU32, SU33, SU34, SU35, SU36, SU37, SU38, SU39, SU39, SU31, SU31, SU32, SU33, SU34, SU35, SU36, SU37, 01, SU6668 and GFB-111; b) compounds that target, reduce or inhibit the activity of fibroblast growth factor receptor (FGFR); c) compounds that target, reduce or inhibit the activity of insulin-like growth factor receptor I (IGF-IR), such as compounds that target, reduce or inhibit the activity of IGF-IR, especially compounds that inhibit the kinase activity of IGF-I receptor, or antibodies that target the extracellular domain of IGF-I receptor or its growth factor; d) compounds that target, reduce or inhibit the activity of Trk receptor tyrosine kinase family, or ephrin B4 inhibitors; e) compounds that target, decrease or inhibit the activity of the AxI family of receptor tyrosine kinases; f) compounds that target, decrease or inhibit the activity of the Ret receptor tyrosine kinase; g) compounds that target, decrease or inhibit the activity of the Kit / SCFR receptor tyrosine kinase, such as imatinib; h) compounds that target, decrease or inhibit the activity of the C-kit receptor tyrosine kinase, which is part of the PDGFR family, such as compounds that target, decrease or inhibit the activity of the C-kit receptor tyrosine kinase family, in particular compounds that inhibit the C-kit receptor, such as imatinib; i) compounds that target, decrease or inhibit the activity of c-Abl family members, their gene fusion products (e.g., BCR-Abl kinase) and mutants, such as compounds that target, decrease or inhibit the activity of c-Abl family members and their gene fusion products, such as N-phenyl-2-pyrimidinyl-amine derivatives, such as imatinib or nilotinib (AMN107); PD180970; AG957; NSC 680410; PD173955 from Parke Davis; or dasatinib (BMS-354825); j) compounds that target, decrease or inhibit the activity of members of the protein kinase C (PKC) and Raf families of serine / threonine kinases, MEK, SRC, JAK / pan-JAK, FAK, PDK1, PKB / Akt, Ras / MAPK, PI3K, SYK, TYK2, BTK and TEC families, and / or members of the cyclin-dependent kinase family (CDKs) (including staurosporine derivatives), such as midostaurin; examples of other compounds include UCN-01, safinol, BAY 43-9006, bryostatin 1, perifosine;llmofosine; RO 318220 and RO 320432; GO 6976; lsis 3521; LY333531 / LY379196; isoquinoline compounds; FTIs; PD184352 or QAN697 (P13K inhibitors) or AT7519 (CDK inhibitors); k) compounds that target, decrease or inhibit the activity of protein tyrosine kinase inhibitors, such as compounds that target, decrease or inhibit the activity of protein tyrosine kinase inhibitors, including imatinib mesylate (Gleevec; TM ) or tyrosine phosphoinhibitors (such as tyrosine phosphoinhibitor A23 / RG-50810); AG 99; tyrosine phosphoinhibitor AG 213; tyrosine phosphoinhibitor AG 1748; tyrosine phosphoinhibitor AG 490; tyrosine phosphoinhibitor B44; tyrosine phosphoinhibitor B44(+) enantiomer; tyrosine phosphoinhibitor AG 555; AG 494; tyrosine phosphoinhibitors AG556, AG957 and adaphostin (4-{[(2,5-dihydroxyphenyl)methyl]amino}-benzoic acid adamantyl ester; NSC680410, adaphost); l) target, decrease or inhibit the epidermal growth factor receptor tyrosine kinase family (EGFR1 Compounds that inhibit the activity of ErbB2, ErbB3, ErbB4 as homodimers or heterodimers) and their mutants, such as compounds that target, reduce or inhibit the activity of the epidermal growth factor receptor family, especially compounds, proteins or antibodies that inhibit members of the EGF receptor tyrosine kinase family, such as EGF receptor, ErbB2, ErbB3 and ErbB4 or bind to EGF or EGF-related ligands, CP358774, ZD 1839, ZM 105180; trastuzumab (Herceptin TM ), Cetuximab (Erbitux TM), Iressa, Tasiva, 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; m) compounds that target, decrease or inhibit the activity of the c-Met receptor, such as compounds that target, decrease or inhibit the activity of c-Met, especially compounds that inhibit the kinase activity of the c-Met receptor, or antibodies that target the extracellular domain of c-Met or bind to HGF; n) compounds that target, decrease or inhibit the kinase activity of 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, buparisib, pictrelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765 and ideelalisib; and q) compounds that target, decrease or inhibit the signaling effects of the hedgehog protein (Hh) or smoothened receptor (SMO) pathway, including but not limited to cyclopamine, vismodegib, itraconazole, erismodegib and IPI-926 (saridegib).

[0421] As used herein, the term "PI3K inhibitor" includes, but is not limited to, compounds having inhibitory activity against one or more enzymes in the phosphatidylinositol-3-kinase family, including, but not limited to, PI3Kα, PI3Kγ, PI3Kδ, PI3Kβ, 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 used in the present invention include, but are not limited to, ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, buparcoxib, pitiracoxib, PF-4691502, BYL-719, datoxib, XL-147, XL-765, and idelacoxib.

[0422] As used herein, the term "Bcl-2 inhibitor" includes, but is not limited to, compounds with inhibitory activity against B-cell lymphoma 2 protein (Bcl-2), including, but not limited to, ABT-199, ABT-731, ABT-737, apogossypol, Ascenta's pan-Bcl-2 inhibitor, curcumin (and its analogs), dual Bcl-2 / Bcl-xL inhibitors (Infinity Pharmaceuticals / Novartis Pharmaceuticals), Genasense (G3139), HA14-1 (and its analogs; see WO2008118802), navitoclax (and its analogs, see US7390799), NH-1 (Shenayng Pharmaceutical University), obatoclax (and its analogs, see WO2004106328), S-001 (Gloria Pharmaceuticals), TW series compounds (Univ. of Michigan), and venetoclax. In some embodiments, the Bcl-2 inhibitor is a small molecule therapeutic agent. In some embodiments, the Bcl-2 inhibitor is a peptidomimetic.

[0423] As used herein, the term "BTK inhibitor" includes, but is not limited to, compounds having inhibitory activity against Bruton's tyrosine kinase (BTK), including, but not limited to, AVL-292 and ibrutinib.

[0424] As used herein, the term "SYK inhibitor" includes, but is not limited to, compounds having inhibitory activity against spleen tyrosine kinase (SYK), including, but not limited to, PRT-062070, R-343, R-333, Excellair, PRT-062607, and fostamatinib.

[0425] Further examples of BTK inhibiting compounds and conditions that can be treated by such compounds in combination with the compounds of the present invention can be found in WO2008039218 and WO2011090760, the entire contents of which are incorporated herein by reference.

[0426] Further examples of SYK inhibiting compounds and conditions that can be treated by such compounds in combination with the compounds of the present invention can be found in WO2003063794, WO2005007623 and WO2006078846, the entire contents of which are incorporated herein by reference.

[0427] Other examples of PI3K inhibitory compounds and conditions that can be treated by such compounds in combination with the compounds of the present invention can be found in WO2004019973, WO2004089925, WO2007016176, US8138347, WO2002088112, WO2007084786, WO2007129161, WO2006122806, WO2005113554 and WO2007044729, the entire contents of which are incorporated herein by reference.

[0428] Further examples of JAK inhibitory compounds and conditions that can be treated by such compounds in combination with the compounds of the present invention can be found in WO2009114512, WO2008109943, WO2007053452, WO2000142246 and WO2007070514, the entire contents of which are incorporated herein by reference.

[0429] Other anti-angiogenic compounds include compounds that have another mechanism of action for their activity, for example, compounds not related to protein or lipid kinase inhibition, such as thalomid TM ) and TNP-470.

[0430] Examples of proteasome inhibitors that can be used in combination with the compounds of the present invention include, but are not limited to, bortezomib, disulfiram, epigallocatechin-3-gallate (EGCG), salinosporin A, carfilzomib, ONX-0912, CEP-18770, and MLN9708.

[0431] Compounds which target, decrease or inhibit the activity of a protein or lipid phosphatase are, for example, inhibitors of phosphatase 1, phosphatase 2A or CDC25, such as okadamic acid or a derivative thereof.

[0432] 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.

[0433] As used herein, the term cyclooxygenase inhibitors includes, but is not limited to, Cox-2 inhibitors, 5-alkyl substituted 2-arylaminophenylacetic acid and derivatives such as celecoxib (Celebrex TM ), rofecoxib (Vioxx TM ), etoricoxib, valdecoxib, or 5-alkyl-2-arylaminophenylacetic acid, such as 5-methyl-2-(2'-chloro-6'-fluoroanilino)phenylacetic acid, lumiracoxib.

[0434] As used herein, the term "bisphosphonates" includes, but is not limited to, etrodronic acid, clodronic acid, tiludronic acid, pamidronic acid, alendronic acid, ibandronic acid, risedronic acid, and zoledronic acid. TM Clodronic acid is sold under the trade name Bonefos TM Tiludronic acid is sold under the brand name Skelid TM Pamidronate is sold under the brand name Aredia TM Alendronate is sold under the brand name Fosamax TM Ibandronic acid is sold under the brand name Bondranat TM Risedronate is sold under the brand name Actonel TM Zoledronic acid is sold under the brand name Zometa TM The term "mTOR inhibitor" relates to compounds that inhibit the mammalian target of rapamycin (mTOR) and have antiproliferative activity, such as sirolimus Everolimus (Certican TM ), CCI-779 and ABT578.

[0435] As used herein, the term "heparanase inhibitor" refers to a compound that targets, reduces, or inhibits the degradation of heparan sulfate. The term includes, but is not limited to, PI-88. As used herein, the term "biological response modifier" refers to a lymphokine or interferon.

[0436] 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, decreases or inhibits the oncogenic activity of Ras; for example, a "farnesyl transferase inhibitor" such as L-744832, DK8G557 or R115777 (Zarnestra TM 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 telomerase activity are particularly compounds that inhibit the telomerase receptor, such as telomestatin.

[0437] As used herein, the term "methionine aminopeptidase inhibitor" refers to a compound that targets, reduces or inhibits the activity of methionine aminopeptidase. Compounds that target, reduce or inhibit the activity of methionine aminopeptidase include, but are not limited to, bengamide or its derivatives.

[0438] As used herein, the term "proteasome inhibitor" refers to a compound that targets, reduces or inhibits the activity of the proteasome. Compounds that target, reduce or inhibit the activity of the proteasome include, but are not limited to, bortezomib (VelcadeTM ) and MLN341.

[0439] As used herein, the term "matrix metalloproteinase inhibitors" or ("MMP" inhibitors) includes, but is not limited to, collagen peptide mimetic and non-peptide mimetic inhibitors, tetracycline derivatives, such as the hydrooxalamidate peptide mimetic inhibitor batimastat and its orally bioavailable analogs marimastat (BB-2516), prinomastat (AG3340), metastat (NSC 683551), BMS-279251, BAY 12-9566, TAA211, MMI270B or AAJ996.

[0440] As used herein, the term "compounds for 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 the FMS-like tyrosine kinase receptor (Flt-3R); interferons, 1-β-D-arabinofuranosylcytosine (ara-c) and busulfan; and ALK inhibitors, which are compounds that target, decrease or inhibit anaplastic lymphoma kinase.

[0441] Compounds that target, decrease or inhibit the activity of the FMS-like tyrosine kinase receptor (Flt-3R) 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.

[0442] 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; compounds that degrade, target, reduce, or inhibit HSP90 client proteins via the ubiquitin proteasome pathway. Compounds that target, reduce, or inhibit the intrinsic ATPase activity of HSP90 include, in particular, compounds, proteins, or antibodies that inhibit the ATPase activity of HSP90, such as 17-allylamino, 17-dimethoxygeldanamycin (17AAG), a geldanamycin derivative; other geldanamycin-related compounds; radicicol; and HDAC inhibitors.

[0443] As used herein, the term "anti-proliferative antibodies" includes, but is not limited to, trastuzumab (Herceptin TM ), trastuzumab-DM1, erbitux, bevacizumab (Avastin TM ), rituximab PRO64553 (anti-CD40) and 2C4 antibodies. Antibodies refer to intact monoclonal antibodies, polyclonal antibodies, multispecific antibodies formed from at least two intact antibodies, and antibody fragments, so long as they exhibit the desired biological activity.

[0444] For the treatment of acute myeloid leukemia (AML), the compounds of the present invention can be combined with standard leukemia therapies, in particular, used in combination with therapies for the treatment of AML. In particular, the compounds of the present invention can be used in combination with, for example, farnesyl transferase inhibitors and / or other drugs that can be used to treat AML, such as daunorubicin, adriamicin (Adriamycin), Ara-C, VP-16, teniposide, mitoxantrone, idarubicin, carboplatin and PKC412.

[0445] Other anti-leukemia compounds include, for example, Ara-C, a pyrimidine analog that is a 2'-α-hydroxyribose (arabinoside) derivative of deoxycytidine. Also included are purine analogs such as hypoxanthine, 6-mercaptopurine (6-MP), and fludarabine phosphate. Compounds that target, reduce, or inhibit the activity of histone deacetylase (HDAC) inhibitors, such as sodium butyrate and suberoylanilide hydroxamic acid (SAHA), inhibit the activity of enzymes called histone deacetylases. Specific HDAC inhibitors include MS275, SAHA, FK228 (formerly FR901228), trichostatin A, and compounds disclosed in US Pat. No. 6,552,065, including but not limited to N-hydroxy-3-[4-[[[2-(2-methyl-1H-indol-3-yl)-ethyl]-amino]methyl]phenyl]-2E-2-acrylamide or a pharmaceutically acceptable salt thereof, and N-hydroxy-3-[4-[(2-hydroxyethyl){2-(1H-indol-3-yl)ethyl]-amino]methyl]phenyl]-2E-2-acrylamide or a pharmaceutically acceptable salt thereof, particularly lactate. As used herein, a somatostatin receptor antagonist refers to a compound that targets, treats, or inhibits somatostatin receptors, such as octreotide and SOM230. Tumor cell destruction methods refer to methods such as ionizing radiation. The term "ionizing radiation" as used herein and hereinafter refers to ionizing radiation in the form of electromagnetic rays (such as X-rays and gamma rays) or particles (such as alpha and beta particles). Ionizing radiation is provided in, but not limited to, radiation therapy and is known in the art. See Hellman, Principles of Radiation Therapy, Cancer, in Principles and Practice of Oncology, Devita et al., eds., 4th ed., Vol. 1, pp. 248-275 (1993).

[0446] Also included are EDG binders and ribonucleotide reductase inhibitors. As used herein, the term "EDG binder" refers to a class of immunosuppressants that regulate lymphocyte recirculation, such as FTY720. The term "ribonucleotide reductase inhibitor" relates to pyrimidine or purine nucleoside analogs, 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 especially hydroxyurea or 2-hydroxy-1H-isoindole-1,3-dione derivatives.

[0447] Also specifically included are those compounds, proteins or monoclonal antibodies to VEGF, such as 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine or a pharmaceutically acceptable salt thereof, 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine succinate; Angiostatin TM ;Endostatin TM ; aminobenzoic acid amide; ZD4190; Zd6474; SU5416; SU6668; bevacizumab; or anti-VEGF antibodies or anti-VEGF receptor antibodies, such as rhuMAb and RHUFab, VEGF aptamers such as Macugon; FLT-4 inhibitors, FLT-3 inhibitors, VEGFR-2 IgGI antibodies, Angiozyme (RPI 4610) and bevacizumab (Avastin TM ).

[0448] As used herein, photodynamic therapy refers to a therapy that uses certain chemicals called photosensitive compounds to treat or prevent cancer. Examples of photodynamic therapy include the use of drugs such as Visudyne TM and porfimer sodium compounds.

[0449] As used herein, angiogenesis-inhibiting steroids refers to compounds that block or inhibit angiogenesis, such as, for example, anecortave, triamcinolone, hydrocortisone, 11-alpha-epihydrocortisol, deoxycortisol, 17alpha-hydroxyprogesterone, corticosterone, deoxycorticosterone, testosterone, estrone, and dexamethasone.

[0450] Implants containing corticosteroids refer to compounds such as fluocinolone and dexamethasone.

[0451] Other chemotherapeutic compounds include, but are not limited to, plant alkaloids, hormone compounds and antagonists; biological response modifiers, preferably lymphokines or interferons; antisense oligonucleotides or oligonucleotide derivatives; shRNA or siRNA; or miscellaneous compounds or compounds with other or unknown mechanisms of action.

[0452] The structures of the active compounds identified by code numbers, generic or trade names can be taken from the actual edition of the standard compendium "The Merck Index" or from databases, for example Patents International (for example IMS World Publications).

[0453] Exemplary Immuno-Oncology Agents

[0454] In some embodiments, the one or more additional therapeutic agents are immuno-oncology agents. As used herein, the term "immuno-oncology agent" refers to an agent that effectively enhances, stimulates, and / or upregulates an immune response in a subject. In some embodiments, administration of an immuno-oncology agent and a compound of the invention has a synergistic effect in treating cancer.

[0455] Immuno-oncology agents can be, for example, small molecule drugs, antibodies, or biologics or small molecules. Examples of biological immuno-oncology agents include, but are not limited to, cancer vaccines, antibodies, and cytokines. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the monoclonal antibody is humanized or human.

[0456] In some embodiments, the immuno-oncology agent is an agonist of (i) stimulatory (including co-stimulatory) receptors or an antagonist of (ii) inhibitory (including co-inhibitory) signals on T cells, both of which enhance antigen-specific T cell responses.

[0457] Certain stimulatory and inhibitory molecules are members of the immunoglobulin superfamily (IgSF). An important family of membrane-bound ligands that bind to costimulatory or co-inhibitory receptors is the B7 family, which includes B7-1, B7-2, B7-H1 (PD-L1), B7-DC (PD-L2), B7-H2 (ICOS-L), B7-H3, B7-H4, B7-H5 (VISTA) and B7-H6. Another family of membrane-bound ligands that bind to costimulatory or co-inhibitory receptors is the TNF molecule family that binds to homologous 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.

[0458] In some embodiments, the immuno-oncology agent is a cytokine that inhibits T cell activation (e.g., IL-6, IL-10, TGF-β, VEGF, and other immunosuppressive cytokines) or a cytokine that stimulates T cell activation for stimulating an immune response.

[0459] In some embodiments, the combination of the compound of the present invention and an immuno-oncology agent can stimulate T cell responses. In some embodiments, the immuno-oncology agent is: (i) an antagonist (e.g., immune checkpoint inhibitor) of a protein that inhibits T cell activation, 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.

[0460] In some embodiments, the immuno-oncology agent is an antagonist of an inhibitory receptor on NK cells or an agonist of an activating receptor on NK cells. In some embodiments, the immuno-oncology agent is an antagonist of KIR, such as lirilumab.

[0461] In some embodiments, the immuno-oncology agent is an agent that inhibits or depletes macrophages or monocytes, including but not limited to CSF-1R antagonists, such as CSF-1R antagonist antibodies including RG7155 (WO11 / 70024, WO11 / 107553, WO11 / 131407, WO13 / 87699, WO13 / 119716, WO13 / 132044) or FPA-008 (WO11 / 140249; WO13169264; WO14 / 036357).

[0462] In some embodiments, the immuno-oncology agent is selected from agonist agents that bind to positive co-stimulatory receptors; blockers that attenuate signaling through inhibitory receptors; antagonists; and one or more agents that systemically increase the frequency of anti-tumor T cells; agents that overcome different immunosuppressive pathways within the tumor microenvironment (e.g., blocking inhibitory receptor engagement (e.g., PD-L1 / PD-1 interaction), depleting or inhibiting Tregs (e.g., using anti-CD25 monoclonal antibodies (e.g., daclizumab) or by ex vivo anti-CD25 bead depletion), inhibiting metabolic enzymes such as IDO, or reversing / preventing T cell energy or exhaustion); and agents that trigger innate immune activation and / or inflammation at the tumor site.

[0463] In some embodiments, the immuno-oncology agent is a CTLA-4 antagonist. In some embodiments, the CTLA-4 antagonist is an antagonist CTLA-4 antibody. In some embodiments, the antagonist CTLA-4 antibody is YERVOY (ipilimumab) or tremelimumab.

[0464] In some embodiments, the immuno-oncology agent is a PD-1 antagonist. In some embodiments, the PD-1 antagonist is administered by infusion. In some embodiments, the immuno-oncology agent is an antibody or antigen-binding portion thereof that specifically binds to the programmed death-1 (PD-1) receptor and inhibits PD-1 activity. In some embodiments, the PD-1 antagonist is an antagonist PD-1 antibody. In some embodiments, the antagonist PD-1 antibody is OPDIVO (nivolumab), KEYTRUDA (pembrolizumab), or MEDI-0680 (AMP-514; WO2012 / 145493). In some embodiments, the immuno-oncology agent may be pidilizumab (CT-011). In some embodiments, the immuno-oncology agent is a recombinant protein consisting of the extracellular domain of PD-L2 (B7-DC) fused to the Fc portion of IgG1, referred to as AMP-224.

[0465] In some embodiments, the immuno-oncology agent is a PD-L1 antagonist. In some embodiments, the PD-L1 antagonist is an antagonist PD-L1 antibody. In some embodiments, the PD-L1 antibody is MPDL3280A (RG7446; WO2010 / 077634), durvalumab (MEDI4736), BMS-936559 (WO2007 / 005874), and MSB0010718C (WO2013 / 79174).

[0466] In some embodiments, the immuno-oncology agent is a LAG-3 antagonist. In some embodiments, the LAG-3 antagonist is an antagonist LAG-3 antibody. In some embodiments, the LAG3 antibody is BMS-986016 (WO10 / 19570, WO14 / 08218), or IMP-731 or IMP-321 (WO08 / 132601, WO009 / 44273).

[0467] In some embodiments, the immuno-oncology agent is a CD137 (4-1BB) agonist. In some embodiments, the CD137 (4-1BB) agonist is an agonist CD137 antibody. In some embodiments, the CD137 antibody is urelumab or PF-05082566 (WO12 / 32433).

[0468] In some embodiments, the immuno-oncology agent is a GITR agonist. In some embodiments, the GITR agonist is an agonistic GITR antibody. In some embodiments, the GITR antibody is BMS-986153, BMS-986156, TRX-518 (WO006 / 105021, WO009 / 009116), or MK-4166 (WO11 / 028683).

[0469] In some embodiments, the immuno-oncology agent is an indoleamine (2,3)-dioxygenase (IDO) antagonist. In some embodiments, the IDO antagonist is selected from epacadostat (INCB024360, Incyte); indoximod (NLG-8189, NewLink Genetics Corporation); capmanitib (INC280, Novartis); GDC-0919 (Genentech / Roche); PF-06840003 (Pfizer); BMS:F001287 (Bristol-Myers Squibb); Phy906 / KD108 (Phytoceutica); an enzyme that breaks down kynurenine (Kynase, Ikena Oncology, formerly known as Kyn Therapeutics); and NLG-919 (WO09 / 73620, WO009 / 1156652, WO11 / 56652, WO12 / 142237).

[0470] In some embodiments, the immuno-oncology agent is an OX40 agonist. In some embodiments, the OX40 agonist is an agonistic OX40 antibody. In some embodiments, the OX40 antibody is MEDI-6383 or MEDI-6469.

[0471] In some embodiments, the immuno-oncology agent is an OX40L antagonist. In some embodiments, the OX40L antagonist is an antagonist OX40 antibody. In some embodiments, the OX40L antagonist is RG-7888 (WO06 / 029879).

[0472] In some embodiments, the immuno-oncology agent is a CD40 agonist. In some embodiments, the CD40 agonist is an agonistic CD40 antibody. In some embodiments, the immuno-oncology agent is a CD40 antagonist. In some embodiments, the CD40 antagonist is an antagonistic CD40 antibody. In some embodiments, the CD40 antibody is lucatumumab or dacetuzumab.

[0473] In some embodiments, the immuno-oncology agent is a CD27 agonist. In some embodiments, the CD27 agonist is an agonistic CD27 antibody. In some embodiments, the CD27 antibody is varlilumab.

[0474] In some embodiments, the immuno-oncology agent is MGA271 (to B7H3) (WO 11 / 109400).

[0475] In some embodiments, the immuno-oncology agent is abagovorizumab, adetumumab, afutuzumab, alemtuzumab, anatumomab mafenatox, apolizumab, atezolizumab, avelumab, blinatumomab, BMS-936559, catumaxomab, durvalumab, epalrestat, epratuzumab, indoximod, inotuzumab, or olivostat. ozogamicin), intetumumab, ipilimumab, isatuximab, lambrolizumab, MED14736, MPDL3280A, nivolumab, obinutuzumab, okatuzumab, ofatumumab, ollatumumab, pembrolizumab, pidilizumab, rituximab, tesimumab, samazumab, or tremelimumab.

[0476] In some embodiments, the immuno-oncology agent is an immunostimulatory agent. For example, antibodies that block the PD-1 and PD-L1 inhibitory axis can release activated tumor-reactive T cells and have been shown in clinical trials to induce durable anti-tumor responses in a growing number of tumor histologies, including some conventional tumor types that have not yet been considered sensitive to immunotherapy. See, for example, Okazaki, T. et al. (2013) Nat. Immunol. 14, 1212–1218; Zou et al. (2016) Sci. Transl. Med. 8. The anti-PD-1 antibody nivolumab ( Bristol-Myers Squibb (also known as ONO-4538, MDX1106, and BMS-936558) has shown the potential to improve overall survival in patients with RCC who have experienced disease progression during or after prior anti-angiogenic therapy.

[0477] In some embodiments, the immunomodulatory therapeutic agent specifically induces apoptosis of tumor cells. Approved immunomodulatory therapeutic agents that can be used in the present invention include pomalidomide ( Celgene; Lenalidomide ( Celgene; Ingenol Methylbutyrate ( LEO Pharma).

[0478] In some embodiments, the immuno-oncology agent is a cancer vaccine. In some embodiments, the cancer vaccine is selected from sipuleucel-T ( Dendreon / Valeant Pharmaceuticals), which is approved for the treatment of asymptomatic or minimally symptomatic metastatic castration-resistant (hormone-refractory) prostate cancer; and talimogene laherparepvec ( BioVex / Amgen, formerly known as T-VEC), a genetically modified oncolytic virus therapy approved for the treatment of unresectable skin, subcutaneous, and lymph node lesions in melanoma. In some embodiments, the immuno-oncology agent is selected from an oncolytic virus therapy such as pexastimogene devaccinepvec (PexaVec / JX-594, SillaJen / formerly Jennerex Biotherapeutics), a thymidine kinase-(TK-) deficient vaccinia virus engineered to express GM-CSF, for use in hepatocellular carcinoma (NCT02562755) and melanoma (NCT00429312); pelareorep ( Oncolytics Biotech, a variant of a respiratory enteric orphan virus (reovirus) that is unable to replicate in RAS-inactivated cells in multiple 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) (NCT 00861627); enadenotucirev (NG-348, PsiOxus, formerly ColoAd1), an adenovirus engineered to express full-length CD80 and an antibody fragment specific for the T-cell receptor CD3 protein, for 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, for melanoma (NCT03003676); and peritoneal disease, colorectal cancer, or ovarian cancer (NCT02963831); GL-ONC1 (GLV-1h68 / GLV-1h153, Genelux GmbH), vaccinia viruses engineered to express β-galactosidase (β-gal) / β-glucuronidase or β-gal / human sodium iodide symporter (hNIS), respectively, have been studied in peritoneal carcinomatosis (NCT01443260); fallopian tube cancer, ovarian cancer (NCT02759588); or CG0070 (Cold Genesys), an adenovirus engineered to express GM-CSF, in bladder cancer (NCT02365818).

[0479] In some embodiments, the immuno-oncology agent is selected from JX-929 (SillaJen / formerly Jennerex Biotherapeutics), a TK and vaccinia growth factor-deficient vaccinia virus engineered to express cytosine deaminase, which is capable of converting the prodrug 5-fluorocytosine to the cytotoxic drug 5-fluorouracil; TG01 and TG02 (Targovax / formerly Oncos), peptide-based immunotherapeutic agents targeting difficult-to-treat RAS mutations; and TILT-123 (TILT Biotherapeutics), an engineered adenovirus designated: Ad5 / 3-E2F-delta24-hTNFα-IRES-hIL20; and VSV-GP (Vira Therapeutics), a vesicular stomatitis virus (VSV) engineered to express the glycoprotein (GP) of lymphocytic choriomeningitis virus (LCMV), which can be further engineered to express a protein designed to elicit antigen-specific CD8 + Antigens to which T cells respond.

[0480] In some embodiments, the immuno-oncology agent is a T cell engineered to express a chimeric antigen receptor or CAR. T cells engineered to express such a chimeric antigen receptor are called CAR-T cells.

[0481] CAR has been constructed to consist of a binding domain (which may 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 internal domain that serves as the functional end of the T cell receptor (TCR), such as the CD3-ζ signaling domain from the TCR, which is capable of generating activation signals in T lymphocytes. Upon antigen binding, such CARs connect to endogenous signaling pathways in effector cells and generate activation signals similar to those initiated by the TCR complex.

[0482] For example, in some embodiments, the CAR-T cell is one of those described in U.S. Pat. No. 8,906,682 (June et al.; hereby incorporated by reference in its entirety), which discloses a CAR-T cell engineered to contain an extracellular domain having an antigen binding domain (such as a domain that binds to CD19) fused to an intracellular signaling domain of the T cell antigen receptor complex ζ chain (such as CD3ζ). When expressed in T cells, the CAR is able to redirect antigen recognition based on antigen binding specificity. In the case of CD19, the antigen is expressed on malignant B cells. Currently, more than 200 clinical trials are underway using CAR-T in a wide range of indications. [https: / / clinicaltrials.gov / ct2 / results?term=chimeric+antigen+receptors&pg=1].

[0483] 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 and the differentiation of innate immune cell subsets expressing IL-17 (such as NK cells). In some embodiments, the activator of RORγt is LYC-55716 (Lycera), which is currently being evaluated in clinical trials for the treatment of solid tumors (NCT02929862).

[0484] In certain embodiments, immunostimulant is an agonist or activator of Toll-like receptor (TLR). Suitable TLR activators include agonists or activators of TLR9, such as SD-101 (Dynavax). SD-101 is a kind of immunostimulatory CpG, which is being studied for B cells, follicular and other lymphomas (NCT02254772). Agonists or activators of TLR8 that can be used in the present invention include Motomod (VTX-2337, VentiRx Pharmaceuticals), which are being studied for head and neck squamous cell carcinoma (NCT02124850) and ovarian cancer (NCT02431559).

[0485] Other immuno-oncology agents that can be used in the present invention include urerulumab (BMS-663513, Bristol-Myers Squibb), an anti-CD137 monoclonal antibody; valirumab (CDX-1127, Celldex Therapeutics), an anti-CD27 monoclonal antibody; BMS-986178 (Bristol-Myers Squibb), an anti-OX40 monoclonal antibody; lirelumab (IPH2102 / BMS-986015, Innate Pharma, Bristol-Myers Squibb), an anti-KIR monoclonal antibody; monalizumab (IPH2201, Innate Pharma, Pharma, AstraZeneca), an anti-NKG2A monoclonal antibody; andecaliximab (GS-5745, Gilead Sciences), an anti-MMP9 antibody; and MK-4166 (Merck & Co.), an anti-GITR monoclonal antibody.

[0486] In some embodiments, the immunostimulatory agent is selected from elotuzumab, mifamurtide, an agonist or activator of a Toll-like receptor, and an activator of RORγt.

[0487] In some embodiments, the immunostimulatory therapeutic agent is recombinant human interleukin 15 (rhIL-15). RhIL-15 has been tested clinically as a therapy for melanoma and renal cell carcinoma (NCT01021059 and NCT01369888) and leukemia (NCT02689453). In some embodiments, the immunostimulatory agent is recombinant human interleukin 12 (rhIL-12). In some embodiments, the IL-15-based immunotherapeutic agent is heterodimeric IL-15 (hetIL-15, Novartis / Admune), which is a fusion complex consisting of a synthetic form of endogenous IL-15 complexed with a soluble IL-15 binding protein IL-15 receptor alpha chain (IL15:sIL-15RA), which has been tested in a Phase 1 clinical trial for 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.

[0488] In some embodiments, the immuno-oncology agent is selected from those described in Jerry L. Adams et al., "Big opportunities for small molecules in immuno-oncology," Cancer Therapy 2015, Vol. 14, pp. 603-622, the contents of which are incorporated herein by reference in their entirety. In some embodiments, the immuno-oncology agent is selected from the examples described in Table 1 of Jerry L. Adams et al. In some embodiments, the immuno-oncology agent is a small molecule that targets an immuno-oncology target selected from those listed in Table 2 of Jerry L. Adams et al. In some embodiments, the immuno-oncology agent is a small molecule agent selected from those listed in Table 2 of Jerry L. Adams et al.

[0489] In some embodiments, the immuno-oncology agent is selected from the small molecule immuno-oncology agents described in Peter L. Toogood, "Small molecule immuno-oncology therapeutic agents," Bioorganic & Medicinal Chemistry Letters 2018, Vol. 28, pp. 319-329, the contents of which are incorporated herein by reference in their entirety. In some embodiments, the immuno-oncology agent is an agent that targets a pathway as described in Peter L. Toogood.

[0490] In some embodiments, the immuno-oncology agent is selected from Sandra L. Ross et al., "Bispecific T cell engager Antibody constructs can mediate bystander tumor cell killing", PLoS ONE 12(8):e0183390, the contents of which are incorporated herein by reference in their entirety. In some embodiments, the immuno-oncology agent is a bispecific T cell engager Antibody Constructs. In some embodiments, bispecific T cell engagers The antibody construct is a CD19 / CD3 bispecific antibody construct. In some embodiments, the bispecific T cell engager The antibody construct is an EGFR / CD3 bispecific antibody construct. In some embodiments, the bispecific T cell engager Antibody constructs activate T cells. In some embodiments, bispecific T cell engagers The antibody construct activates T cells, which release cytokines that induce upregulation of intercellular adhesion molecule 1 (ICAM-1) and FAS on bystander cells. In some embodiments, the bispecific T cell engager The antibody construct activates T cells, which results in induced bystander cell lysis. In some embodiments, the bystander cells are located in a solid tumor. In some embodiments, the lysed bystander cells are located in the vicinity of -activated T cells. In some embodiments, bystander cells include tumor-associated antigen (TAA) negative cancer cells. In some embodiments, bystander cells include EGFR negative cancer cells. In some embodiments, the immuno-oncology agent is an antibody that blocks the PD-L1 / PD1 axis and / or CTLA4. In some embodiments, the immuno-oncology agent is a tumor-infiltrating T cell that is expanded ex vivo. In some embodiments, the immuno-oncology agent is a bispecific antibody construct or a chimeric antigen receptor (CAR) that directly connects T cells to tumor-associated surface antigens (TAAs).

[0491] Exemplary immune checkpoint inhibitors

[0492] In some embodiments, the immuno-oncology agent is an immune checkpoint inhibitor as described herein.

[0493] As used herein, the term "checkpoint inhibitor" refers to agents that can be used to prevent cancer cells from evading the patient's immune system. One of the main mechanisms of anti-tumor immune destruction is called "T cell exhaustion," which is caused by long-term exposure to antigens that lead to upregulation of inhibitory receptors. These inhibitory receptors act as immune checkpoints to prevent uncontrolled immune responses.

[0494] PD-1 and co-inhibitory receptors such as cytotoxic T lymphocyte antigen 4 (CTLA-4), B and T lymphocyte attenuator (BTLA; CD272), T cell immunoglobulin and mucin domain-containing 3 (Tim-3), and lymphocyte activation gene-3 (Lag-3; CD223) are commonly referred to as checkpoint regulators. They act as molecular "gatekeepers," allowing extracellular information to determine whether cell cycle progression and other intracellular signaling processes should proceed.

[0495] In some embodiments, immune checkpoint inhibitors are antibodies directed against PD-1, which binds to the programmed cell death 1 receptor (PD-1) to prevent the receptor from binding to its inhibitory ligand, PDL-1, thereby overriding the tumor's ability to suppress the host's anti-tumor immune response.

[0496] In one aspect, the checkpoint inhibitor is a biotherapeutic agent or a small molecule. In another aspect, the checkpoint inhibitor is a monoclonal antibody, a humanized antibody, a fully human antibody, a fusion protein, or a combination thereof. In other aspects, the checkpoint inhibitor inhibits a checkpoint protein selected from the following items: CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK 1, CHK2, A2aR, B-7 family ligand, or a combination thereof. In another aspect, the check point inhibitor interacts with a ligand selected from the check point proteins of the following items: 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 ligands or a combination thereof. In one aspect, the check point inhibitor is an immunostimulant, a T cell growth factor, an interleukin, an antibody, a vaccine or a combination thereof. In other aspects, the interleukin is IL-7 or IL-15. In specific aspects, the interleukin is a glycosylated IL-7. In another aspect, the vaccine is a dendritic cell (DC) vaccine.

[0497] Check point inhibitors include any agent that blocks or suppresses the inhibitory pathway of the immune system in a statistically significant manner.Such inhibitors may include small molecule inhibitors or may include antibodies or their antigen binding fragments that bind to and block or suppress immune checkpoint receptors, or antibodies that bind to and block or suppress immune checkpoint receptor ligands. Exemplary checkpoint molecules that can be targeted for blocking or inhibition include but are not limited to CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, GAL9, LAG3, TIM3, VISTA, KIR, 2B4 (belonging to the CD2 family of molecules and in all NK, γδ and memory CD8 +(αβ) T cells), CD160 (also known as BY55), CGEN-15049, CHK 1 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, CD 160, and CGEN-15049. Exemplary immune checkpoint inhibitors include, but are not limited to, tremelimumab (CTLA-4 blocking antibody), anti-OX40, PD-L1 monoclonal antibody (anti-B7-H1; MEDI4736), MK-3475 (PD-1 blocker), nivolumab (anti-PD1 antibody), CT-011 (anti-PD1 antibody), BY55 monoclonal antibody, AMP224 (anti-PDL1 antibody), BMS-936559 (anti-PDL1 antibody), MPLDL3280A (anti-PDL1 antibody), MSB0010718C (anti-PDL1 antibody) and ipilimumab (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.

[0498] 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 nivolumab. Ipilimumab and pembrolizumab In some embodiments, the checkpoint inhibitor is selected from nivolumab (anti-PD-1 antibody, Bristol-Myers Squibb; pembrolizumab (anti-PD-1 antibody, Merck); Ipilimumab (anti-CTLA-4 antibody, Bristol-Myers Squibb; durvalumab (anti-PD-L1 antibody, AstraZeneca); and atezolizumab (anti-PD-L1 antibody, Genentech).

[0499] 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, lirelumab, IPH2101, pembrolizumab and tremelimumab group.

[0500] In some embodiments, the immune checkpoint inhibitor is REGN2810 (Regeneron), an anti-PD-1 antibody being tested 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 and is in clinical trials for diffuse large B-cell lymphoma and multiple myeloma; avelumab ( Pfizer / Merck KGaA), also known as MSB0010718C), a fully human IgG1 anti-PD-L1 antibody in clinical trials for non-small cell lung cancer, Merkel cell carcinoma, mesothelioma, solid tumors, renal cancer, ovarian cancer, bladder cancer, head and neck cancer, and gastric cancer; or PDR001 (Novartis), an inhibitory antibody that binds to PD-1 in clinical trials for non-small cell lung cancer, melanoma, triple-negative breast cancer, and advanced or metastatic solid tumors. Tremelimumab (CP-675,206; Astrazeneca) is a fully human monoclonal antibody against CTLA-4 that is being studied in clinical trials for multiple indications, including mesothelioma, colorectal cancer, renal cancer, breast cancer, lung cancer, non-small cell lung cancer, pancreatic ductal adenocarcinoma, pancreatic cancer, germ cell cancer, head and neck squamous cell carcinoma, hepatocellular carcinoma, prostate cancer, endometrial cancer, metastatic liver cancer, hepatocellular carcinoma, large B-cell lymphoma, ovarian cancer, cervical cancer, metastatic undifferentiated 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 for advanced solid tumors (NCT02694822).

[0501] In some embodiments, the checkpoint inhibitor is an inhibitor of T cell immunoglobulin mucin-containing protein-3 (TIM-3). TIM-3 inhibitors that can be used in the present invention include TSR-022, LY3321367, and MBG453. TSR-022 (Tesaro) is an anti-TIM-3 antibody being studied in solid tumors (NCT02817633). LY3321367 (Eli Lilly) is an anti-TIM-3 antibody being studied in solid tumors (NCT03099109). MBG453 (Novartis) is an anti-TIM-3 antibody being studied in advanced malignancies (NCT02608268).

[0502] In some embodiments, checkpoint inhibitors are inhibitors of T cell immunoreceptors with Ig and ITIM domains or TIGIT (immunoreceptors 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 antibodies (NCT02913313); OMP-313M32 (Oncomed); and anti-TIGIT monoclonal antibodies (NCT03119428).

[0503] 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) is an anti-LAG-3 antibody being studied in glioblastoma and gliosarcoma (NCT02658981). REGN3767 (Regeneron) is also an anti-LAG-3 antibody being studied in malignant tumors (NCT03005782). IMP321 (Immutep SA) is a LAG-3-Ig fusion protein being studied in melanoma (NCT02676869); adenocarcinoma (NCT02614833); and metastatic breast cancer (NCT00349934).

[0504] Checkpoint inhibitors that can be used in the present invention include OX40 agonists. OX40 agonists being studied in clinical trials include PF-04518600 / PF-8600 (Pfizer), an agonistic anti-OX40 antibody for metastatic renal cancer (NCT03092856) and advanced cancers and tumors (NCT02554812; NCT05082566); GSK3174998 (Merck), an agonistic anti-OX40 antibody in a Phase 1 cancer trial (NCT02528357); MEDI0562 (Medimmune / AstraZeneca) , an agonist anti-OX40 antibody, for advanced solid tumors (NCT02318394 and NCT02705482); MEDI6469, an agonist anti-OX40 antibody (Medimmune / AstraZeneca), for patients with colorectal cancer (NCT02559024), breast cancer (NCT01862900), head and neck cancer (NCT02274155), and metastatic prostate cancer (NCT01303705); and BMS-986178 (Bristol-Myers Squibb), an agonist anti-OX40 antibody, for advanced cancers (NCT02737475).

[0505] Checkpoint inhibitors useful in the present invention include CD137 (also known as 4-1BB) agonists. CD137 agonists being studied in clinical trials include utomilumab (PF-05082566, Pfizer), an agonistic anti-CD137 antibody being studied for diffuse large B-cell lymphoma (NCT02951156) and for advanced cancers and tumors (NCT02554812 and NCT05082566); urerulumab (BMS-663513, Bristol-Myers Squibb), an agonistic anti-CD137 antibody being studied for melanoma and skin cancer (NCT02652455) and for glioblastoma and gliosarcoma (NCT02658981); and CTX-471 (Compass Therapeutics), an agonistic anti-CD137 antibody being studied for metastatic or locally advanced malignancies (NCT03881488).

[0506] Checkpoint inhibitors that can be used in the present invention include CD27 agonists. CD27 agonists being studied in clinical trials include valirumab (CDX-1127, Celldex Therapeutics), an agonist anti-CD27 antibody, for squamous cell head and neck cancer, ovarian tumors, colorectal cancer, renal cell carcinoma, and glioblastoma (NCT02335918); lymphoma (NCT01460134); and glioma and astrocytoma (NCT02924038).

[0507] Checkpoint inhibitors that can be used in the present invention include glucocorticoid-induced tumor necrosis factor receptor (GITR) agonists. GITR agonists being studied in clinical trials include TRX518 (Leap Therapeutics), an agonist anti-GITR antibody for malignant melanoma and other solid tumors (NCT01239134 and NCT02628574); GWN323 (Novartis), an agonist anti-GITR antibody for solid tumors and lymphomas (NCT 02740270); INCAGN01876 (Incyte / Agenus), an agonist anti-GITR antibody for advanced cancers (NCT02697591 and NCT03126110); MK-4166 (Merck), an agonist anti-GITR antibody for solid tumors (NCT02132754) and MEDI1873 (Medimmune / AstraZeneca), a human IgG1 An agonistic hexameric GITR ligand with an Fc domain for advanced solid tumors (NCT02583165).

[0508] Checkpoint inhibitors that can be used in the present invention include inducible T cell co-stimulator (ICOS, also known as CD278) agonists. ICOS agonists currently under investigation in clinical trials include MEDI-570 (Medimmune), an agonistic anti-ICOS antibody for lymphoma (NCT02520791); GSK3359609 (Merck), an agonistic anti-ICOS antibody in Phase 1 (NCT02723955); and JTX-2011 (Jounce Therapeutics), an agonistic anti-ICOS antibody in Phase 1 (NCT02904226).

[0509] Checkpoint inhibitors useful in the present invention include killer IgG-like receptor (KIR) inhibitors. KIR inhibitors being studied in clinical trials include lirelumab (IPH2102 / BMS-986015, Innate Pharma / Bristol-Myers Squibb), an anti-KIR antibody, for leukemias (NCT01687387, NCT02399917, NCT02481297, NCT02599649), multiple myeloma (NCT02252263), and lymphoma (NCT01592370); IPH2101 (1-7F9, Innate Pharma), for myeloma (NCT01222286 and NCT01217203); and IPH4102 (Innate Pharma), an anti-KIR antibody that binds to three domains of the long cytoplasmic tail (KIR3DL2), for lymphoma (NCT02593045).

[0510] Checkpoint inhibitors useful in the present invention include CD47 inhibitors of the interaction between CD47 and signal regulatory protein alpha (SIRPα). CD47 / SIRPa inhibitors being studied in clinical trials include ALX-148 (Alexo Therapeutics), an antagonist variant of (SIRPa) that binds to CD47 and blocks CD47 / SIRPa-mediated signaling, in Phase 1 (NCT03013218); TTI-621 (SIRPa-Fc, Trillium Therapeutics), a soluble recombinant fusion protein generated 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 transmitting a “don’t eat” signal to macrophages, in Phase 1 clinical trials (NCT02890368 and NCT02663518); CC-90002 (Celgene), an anti-CD47 antibody for leukemia (NCT02641002); and Hu5F9-G4 (Forty Seven, Inc.), for colorectal and solid tumors (NCT02953782), acute myeloid leukemia (NCT02678338), and lymphoma (NCT02953509).

[0511] Checkpoint inhibitors that can be used in the present invention include CD73 inhibitors. CD73 inhibitors being studied in clinical trials include MEDI9447 (Medimmune), an anti-CD73 antibody for solid tumors (NCT02503774); and BMS-986179 (Bristol-Myers Squibb), an anti-CD73 antibody for solid tumors (NCT02754141).

[0512] Checkpoint inhibitors that can be used in the present invention include agonists of the stimulator of interferon genes protein (STING, also known as transmembrane protein 173 or TMEM173). Agonists of STING being studied in clinical trials include MK-1454 (Merck), an agonist synthetic cyclic dinucleotide for lymphoma (NCT03010176); and ADU-S100 (MIW815, Aduro Biotech / Novartis), an agonist synthetic cyclic dinucleotide, in Phase 1 (NCT02675439 and NCT03172936).

[0513] Checkpoint inhibitors that can be used in the present invention include CSF1R inhibitors. CSF1R inhibitors being studied in clinical trials include pexidartinib (PLX3397, Plexxikon), a small molecule inhibitor of CSF1R for colorectal cancer, pancreatic cancer, metastatic and advanced cancer (NCT02777710) and melanoma, non-small cell lung cancer, squamous cell head and neck cancer, gastrointestinal stromal tumor (GIST) and ovarian cancer (NCT02452424); and IMC-CS4 (LY3022855, Lil ly), an anti-CSF-1R antibody, in development for pancreatic cancer (NCT03153410), melanoma (NCT03101254), and solid tumors (NCT02718911); and BLZ945 (4-[2((1R,2R)-2-hydroxycyclohexylamino)-benzothiazol-6-yloxy]-pyridine-2-carboxylic acid carboxamide, Novartis), an orally available CSF1R inhibitor, in development for advanced solid tumors (NCT02829723).

[0514] Checkpoint inhibitors that can be used in the present invention include NKG2A receptor inhibitors. NKG2A receptor inhibitors currently being studied in clinical trials include monalidomide (IPH2201, Innate Pharma), an anti-NKG2A antibody for head and neck cancer (NCT02643550) and chronic lymphocytic leukemia (NCT02557516).

[0515] In some embodiments, the immune checkpoint inhibitor is selected from nivolumab, pembrolizumab, ipilimumab, avelumab, durvalumab, atezolizumab, or pidilizumab.

[0516] Examples

[0517] The following examples are intended to illustrate the present invention and should not be construed as limiting thereon. Unless otherwise stated, one or more tautomeric forms of the compounds of the examples described below can be prepared and / or separated in situ. All tautomeric forms of the compounds of the examples described below should be considered to be disclosed. Temperatures are expressed in degrees Celsius. If not otherwise mentioned, all evaporations are under reduced pressure, preferably between about 15mm Hg and 100mm Hg (=20-133mbar). The structures of the final products, intermediates, and starting materials are confirmed by standard analytical methods, such as microanalysis and spectral characteristics, such as MS, IR, NMR. The abbreviations used are conventional abbreviations in this area.

[0518] All starting materials, structural units, reagents, acids, bases, dehydrating agents, solvents and catalysts for synthesizing the compounds of this invention are commercially available or can be prepared by organic synthesis methods known to those of ordinary skill in the art. In addition, the compounds of the present invention can be produced by organic synthesis methods known to those of ordinary skill in the art, as shown in the following examples.

[0519] Example 1: Synthesis of exemplary compounds

[0520] I-1

[0521]

[0522] Step 1 :(2E)-3-[4-amino-2-(methylthio)pyrimidin-5-yl]prop-2-enoic acid ethyl ester

[0523] To a stirred solution of 4-amino-2-(methylsulfanyl)pyrimidine-5-carboxaldehyde (10 g, 59 mmol, 1 equivalent) in THF (150 mL) was added ethyl 2-(triphenyl-λ5-phosphinyl)acetate (22.65 g, 65 mmol, 1.1 equivalents) at room temperature. The resulting mixture was stirred at 65 ° C for 3 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% MeCN / water (0.1% TFA) in 10 min with a UV detector (254 nm) to produce (2E)-3-[4-amino-2-(methylsulfanyl)pyrimidin-5-yl]prop-2-enoic acid ethyl ester (9.0 g, 63.6%) as a light yellow solid. LC-MS (ESI) m / z, 240.2 [M + H]

[0524] Step 2: 2-(Methylthio)-8H-pyrido[2,3-d]pyrimidin-7-one

[0525] To a stirred solution of (2E)-3-[4-amino-2-(methylsulfanyl)pyrimidin-5-yl]prop-2-enoic acid ethyl ester (10 g, 41.79 mmol, 1 equiv) in DIEA (100 mL) was added DBU (9.54 g, 62.68 mmol, 1.5 equiv) at room temperature. The resulting mixture was stirred at 130 ° C for 3 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% MeCN / water (0.1% FA) in 15 min with a UV detector (254 nm) to produce 2-(methylsulfanyl)-8H-pyrido[2,3-d]pyrimidin-7-one (5.5 g, 68.11%) as a light yellow solid. LC-MS (ESI) m / z 194.2 [M+H]

[0526] Step 3: 2-Chloro-8H-pyrido[2,3-d]pyrimidin-7-one

[0527] A solution of 2-(methylthio)-8H-pyrido[2,3-d]pyrimidin-7-one (6 g, 31.053 mmol, 1 equiv) in SO2Cl2 (60 mL) was stirred at 50 ° C for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% MeCN / water (0.1% FA) in 10 min with a UV detector (254 nm) to produce 2-chloro-8H-pyrido[2,3-d]pyrimidin-7-one (3 g, 53.21%) as an off-white solid. LC-MS (ESI) m / z 182.5 [M+H]

[0528] Step 4: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8H-pyrido[2,3-d]pyrimidin-7-one

[0529] To a stirred solution of 2-chloro-8H-pyrido[2,3-d]pyrimidin-7-one (3 g, 16.522 mmol, 1 eq) and 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (4.81 g, 24.7 mmol, 1.5 eq) in 1,4-dioxane (60 mL) and H₂O (12 mL) was added Pd(dppf)Cl₂ (2.42 g, 3.3 mmol, 0.2 eq) and K₃PO₄ (8.77 g, 41.305 mmol, 2.5 eq) at room temperature. The resulting mixture was stirred at 90° C. under a nitrogen atmosphere for 2 hours. The resulting mixture was filtered, and the filter cake was washed with 1,4-dioxane (3*50 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, mobile phase, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8H-pyrido[2,3-d]pyrimidin-7-one (2.3 g, 47.14%) as an off-white solid. LC-MS (ESI) m / z 296.3 [M+H]

[0530] Step 5: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one

[0531] To a stirred solution of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8H-pyrido[2,3-d]pyrimidin-7-one (1.9 g, 6.4 mmol, 1.1 equiv) and 2-[4-(chloromethyl)phenyl]-1-methyl-4-(trifluoromethyl)imidazole (1.61 g, 5.8 mmol, 1 equiv) in DMF (20 mL) was added 1,1,3,3-tetramethylguanidine (1.01 g, 8.774 mmol, 1.5 equiv) at room temperature. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 3 h. The resulting mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with water (3 x 100 mL), dried over anhydrous NaSO, and concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN in water (0.1% FA) over 20 min with UV detector (254 nm) to give 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one (2.16 g, 69.22%) as an off-white solid. 1H NMR (400MHz, DMSO-d6) δ9.29(s,1H),8.69(s,1H),8.16(d,J=9.6Hz,1H),7.91(d,J=1.4Hz,1H),7.68–7.58(m,2H),7.4 0(d,J=8.2Hz,2H),6.90(d,J=9.5Hz,1H),5.57(s,2H),3.78(d,J=36.0Hz,6H),1.72(m,1H),1.01(m,2H),0.77(m,2H). LC-MS(ESI)m / z 534.0[M+H]

[0532] I-2

[0533]

[0534] Step 1: 4-Amino-2-chloropyrimidin-5-ol

[0535] To a stirred mixture of 2-chloro-5-methoxypyrimidine-4-amine (1 g, 6.27 mmol, 1 equivalent) in CHCl (10 mL, 157.306 mmol, 25.10 equivalents) was added BBr (9.1 mL, 96.26 mmol, 15.36 equivalents) at room temperature. The resulting mixture was stirred at room temperature overnight. The reaction was quenched by adding MeOH (4 mL) at 0 ° C. The precipitated solid was collected by filtration and washed with MeOH (3x5 mL). The resulting solid was dried under vacuum to obtain 4-amino-2-chloropyrimidine-5-ol (0.7 g, 76.74%) as a white solid. The crude product mixture was used directly in the next step without further purification. LC-MS (ESI) m / z 146 [M+H].

[0536] Step 2: 2-Chloro-5-(2-chloroethoxy)pyrimidin-4-amine

[0537] To a stirred solution of 4-amino-2-chloropyrimidin-5-ol (732 mg, 5.029 mmol, 1 eq) and KCO (4170.36 mg, 30.174 mmol, 6 eq) in DMF (10 ml, 129.215 mmol, 25.69 eq) was added 1-bromo-2-chloroethane (2163.71 mg, 15.087 mmol, 3 eq) dropwise under a nitrogen atmosphere at room temperature. The final reaction mixture was stirred at room temperature overnight. The resulting mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (3 x 10 mL), dried over anhydrous NaSO, filtered and concentrated under reduced pressure to give 2-chloro-5-(2-chloroethoxy)pyrimidin-4-amine (450 mg, 43.01%) as an off-white solid. The crude product mixture was used directly in the next step without further purification. LC-MS (ESI) m / z 208 [M+H]

[0538] Step 3: 2-Chloro-6H,7H,8H-pyrimido[5,4-b][1,4]oxazine

[0539] To a stirred solution of 2-chloro-5-(2-chloroethoxy)pyrimidin-4-amine (450 mg, 2.163 mmol, 1 eq) in DMF (5 ml, 64.608 mmol, 24.63 eq) was added KCO (1195.77 mg, 8.652 mmol, 4 eq) in portions at room temperature under a nitrogen atmosphere. The final reaction mixture was stirred at 100 ° C overnight. The resulting mixture was extracted with EtOAc (3x20 mL). The combined organic layers were washed with brine (3x10 mL), dried over anhydrous NaSO, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE / EA (5:1)) to give 2-chloro-6H, 7H, 8H-pyrimido[5,4-b][1,4]oxazine (300 mg, 80.83%) as an off-white solid. LC-MS (ESI) m / z 172 [M+H]

[0540] Step 4: 2-[4-({2-chloro-6H,7H-pyrimido[5,4-b][1,4]oxazin-8-yl}methyl)phenyl]-1-methyl-4-(trifluoromethyl)imidazole

[0541] To a stirred solution of 2-chloro-6H,7H,8H-pyrimido[5,4-b][1,4]oxazine (280 mg, 1.632 mmol, 2.5 equiv) and 2-[4-(chloromethyl)phenyl]-1-methyl-4-(trifluoromethyl)imidazole (179.29 mg, 0.653 mmol, 1 equiv) in DMF (5 ml, 64.608 mmol, 39.59 equiv) was added KCO (270.64 mg, 1.958 mmol, 3 equiv) in portions at room temperature under a nitrogen atmosphere. The final reaction mixture was stirred at 45 ° C for 3 h. The reaction was quenched with water at 0 ° C. The resulting mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (1 x 10 mL), dried over anhydrous NaSO, filtered and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 10% to 100% MeCN / water (10 mmol / L NH4HCO3) over 10 min with UV detector (254 nm) to yield 2-[4-({2-chloro-6H,7H-pyrimido[5,4-b][1,4]oxazin-8-yl}methyl)phenyl]-1-methyl-4-(trifluoromethyl)imidazole (226.1 mg, 84.52%) as an orange solid. LC-MS (ESI) m / z 410 [M+H]

[0542] Step 5: 4-Cyclopropyl-6-methoxy-5-[8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-6H,7H-pyrimido[5,4-b][1,4]oxazin-2-yl]pyrimidine

[0543] To a stirred solution of 2-[4-({2-chloro-6H,7H-pyrimido[5,4-b][1,4]oxazin-8-yl}methyl)phenyl]-1-methyl-4-(trifluoromethyl)imidazole (160 mg, 0.390 mmol, 1 eq) and 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (113.62 mg, 0.585 mmol, 1.5 eq) in 1,4-dioxane (3 ml, 12.000 mmol, 30.74 eq) was added K3PO4 (207.19 mg, 0.975 mmol, 2.5 eq) and HO (0.3 mL, 16.653 mmol, 42.65 eq) at room temperature under nitrogen atmosphere. To the above mixture was added Pd(dppf)Cl2 (57.14 mg, 0.078 mmol, 0.2 eq) at room temperature. The resulting mixture was stirred at 90 ° C for another 2h. The reaction was quenched with water at 0 ° C. The resulting mixture was extracted with EtOAc (3x30mL). The combined organic layer was washed with brine (1x10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% MeCN / water (0.1% FA) in 10min with a UV detector (254nm) to produce 4-cyclopropyl-6-methoxy-5-[8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-6H, 7H-pyrimido[5,4-b][1,4]oxazine-2-yl]pyrimidine (76.3mg, 37.33%) as an off-white solid.

[0544] 1 H NMR(400MHz, DMSO-d6)δ8.66(s,1H),8.15(s,1H),7.94(d,J=1.4Hz,1H),7.72–7.65(m,2H),7.46(d,J=8.1Hz,2H),4 .96(s,2H),4.35(s,3H),3.88(s,3H),3.77(s,3H),3.69(s,2H),1.86(s,1H),1.05–0.98(m,2H),0.89–0.81(m,2H). LC-MS(ESI)m / z524.20[M+H]

[0545] I-3

[0546]

[0547] I-3 was synthesized in a similar manner to I-2.

[0548] 1H NMR (400 MHz, chloroform-d) δ 8.72 (s, 1H), 8.19 (s, 1H), 7.57 (s, 2H), 7.47 (s, 1H), 7.41 (s, 2H), 5.11 (s, 2H), 4.53 (s, 1H), 4.36 (s, 2H), 4.07 (s, 3H), 3.72 (s, 2H), 1.93 (s, 1H), 1.49 (d, J = 5.5 Hz, 6H), 1.34 (s, 2H), 0.95 (s, 2H). LC-MS (ESI) m / z 552.40 [M+H]

[0549] I-4

[0550]

[0551] Step 1: 8-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-2-(methylthio)pyrido[2,3-d]pyrimidin-7-one

[0552] To a stirred solution of 2-(methylthio)-8H-pyrido[2,3-d]pyrimidin-7-one (200 mg, 1.035 mmol, 1 equiv) and 2-[4-(chloromethyl)phenyl]-1-isopropyl-4-(trifluoromethyl)imidazole (313.35 mg, 1.035 mmol, 1 equiv) in DMF (2 mL) was added 1,1,3,3-tetramethylguanidine (178.83 mg, 1.552 mmol, 1.5 equiv) dropwise at room temperature. The resulting mixture was stirred at room temperature overnight under a nitrogen atmosphere. The reaction was quenched with water (10 mL) at room temperature. The resulting mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (3 x 10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The crude product was used directly in the next step without further purification. LC-MS (ESI) m / z: 460.1 [M+H]

[0553] Step 2 2-Chloro-8-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one

[0554] To a stirred solution of 8-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-2-(methylthio)pyrido[2,3-d]pyrimidin-7-one (150 mg, 0.326 mmol, 1 eq) in CHCl (1 mL) and EtOH (0.1 mL) was added SOCl (132.17 mg, 0.978 mmol, 3 eq) dropwise at room temperature. The resulting mixture was stirred at room temperature for 2 h and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 10% to 100% MeCN / water (0.1% NH 3 · H 2 O) over 10 min with UV detection (254 nm) to afford 2-chloro-8-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one (100 mg, 68.40%) as a light brown solid. LC-MS (ESI) m / z: 448.1 [M+H]

[0555] Step 3: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one

[0556] To a stirred solution of 2-chloro-8-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one (100 mg, 0.223 mmol, 1 eq) and 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (64.98 mg, 0.335 mmol, 1.5 eq) in 1,4-dioxane (0.8 mL) and water (0.2 mL) was added Pd(dppf)Cl2 (32.68 mg, 0.045 mmol, 0.2 eq) and K3PO4 (118.49 mg, 0.557 mmol, 2.5 eq) at room temperature. The resulting mixture was stirred at 90 °C under a nitrogen atmosphere for 2 h. The resulting mixture was filtered; the filter cake was washed with 1,4-dioxane (3 x 15 mL). The filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC under the following conditions (column: YMC-Actus Triart C18ExRS, 30*150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 55% B to 65% B, 65% B in 7 min; wavelength: 220 nm; RT1 (min): 5.03) to give 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one (26.1 mg, 20.42%) as a brown liquid. 1 H NMR (400MHz, chloroform-d) δ9.02(s,1H),8.69(s,1H),7.77(d,J=9.5Hz,1H),7.61(d,J=8.0Hz,2H),7.47–7.36(m,3H),6.87(d,J=9.5 Hz,1H),5.71(s,2H),4.51(p,J=6.7Hz,1H),3.93(s,3H),1.70(m,1H),1.42(d,J=6.7Hz,6H),1.30–1.15(m,2H),0.86(m,2H). LC-MS(ESI)m / z562.3[M+H].

[0557] I-5

[0558]

[0559] Step 1: Synthesis of ethyl 2-[(4-amino-2-chloropyrimidin-5-yl)oxy]acetate

[0560] To a stirred solution of 4-amino-2-chloropyrimidin-5-ol (511 mg, 3.511 mmol, 1 eq) and KCO (345 mg, 2.496 mmol, 0.71 eq) in DMF (10 mL, 73.095 mmol, 36.80 eq) was added ethyl bromoacetate (383 mg, 2.293 mmol, 0.65 eq) dropwise under a nitrogen atmosphere at room temperature. The resulting mixture was stirred at room temperature overnight. The resulting mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (3 x 10 mL), dried over anhydrous NaSO, filtered and concentrated under reduced pressure. The resulting mixture was concentrated under vacuum to give ethyl 2-[(4-amino-2-chloropyrimidin-5-yl)oxy]acetate (461.0 mg, 56.69%) as an off-white solid. The crude product mixture was used directly in the next step without further purification. LC-MS (ESI) m / z 232 [M+H]

[0561] Step 2: 2-Chloro-6H,8H-pyrimido[5,4-b][1,4]oxazin-7-one

[0562] To a stirred solution of ethyl 2-[(4-amino-2-chloropyrimidin-5-yl)oxy]acetate (465 mg, 2.007 mmol, 1 eq) in DMF (8 mL, 103.372 mmol, 51.49 eq) was added KCO (138.72 mg, 1.004 mmol, 0.5 eq) in portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 60 ° C for another overnight. The resulting mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (3 x 10 mL), dried over anhydrous NaSO, filtered and concentrated under reduced pressure. The resulting mixture was concentrated under vacuum to give 2-chloro-6H, 8H-pyrimido[5,4-b][1,4]oxazin-7-one (214.9 mg, 57.69%) as a yellow solid. The crude product mixture was used directly in the next step without further purification. LC-MS (ESI) m / z 186 [M+H]

[0563] Step 3: 2-Chloro-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-6H-pyrimido[5,4-b][1,4]oxazin-7-one

[0564] To a stirred solution of 2-chloro-6H,8H-pyrimido[5,4-b][1,4]oxazin-7-one (215 mg, 1.159 mmol, 1 eq) and KCO (160 mg, 1.158 mmol, 1.00 eq) in DMF (4 mL, 51.686 mmol, 44.61 eq) was added 2-[4-(chloromethyl)phenyl]-1-methyl-4-(trifluoromethyl)imidazole (106 mg, 0.386 mmol, 0.33 eq) dropwise under a nitrogen atmosphere at room temperature. The resulting mixture was stirred at 50 ° C for 3 h. The resulting mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (3 x 10 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE / EA (1:1)) to give 2-chloro-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-6H-pyrimido[5,4-b][1,4]oxazin-7-one (125 mg, 25.46%) as an off-white solid. LC-MS (ESI) m / z 424 [M+H]

[0565] Step 4: Synthesis of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-6H-pyrimido[5,4-b][1,4]oxazin-7-one

[0566] To a stirred solution of 2-chloro-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-6H-pyrimido[5,4-b][1,4]oxazin-7-one (130 mg, 0.307 mmol, 1 eq) and 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (89.27 mg, 0.461 mmol, 1.5 eq) in 1,4-dioxane (2 mL, 8.000 mmol, 26.08 eq) was added K3PO4 (162.79 mg, 0.767 mmol, 2.5 eq) and HO (0.18 mL, 9.992 mmol, 32.57 eq) at room temperature under nitrogen atmosphere. To the above mixture was added Pd(dppf)Cl2 (44.89 mg, 0.061 mmol, 0.2 eq) at room temperature. The resulting mixture was stirred at 90 ° C for 3h. The resulting mixture was extracted with EtOAc (3x 20mL). The combined organic layer was washed with brine (3x10mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% MeCN / water (0.1% FA) in 10min with UV detector (254nm) to produce 2- (4-cyclopropyl-6-methoxypyrimidin-5-yl) -8- ({4- [1- methyl -4- (trifluoromethyl) imidazoles -2-yl] phenyl} methyl) -6H- pyrimido [5,4-b] [1,4] oxazine -7- one (80mg, 48.52%) as an off-white solid. 1 H NMR (400MHz, DMSO-d6) δ8.64(s,1H),8.53(s,1H),7.92(d,J=1.4Hz,1H),7.68–7.61(m,2H),7.46(d,J=8.2Hz,2H),5.24(s ,2H),5.05(s,2H),3.81(s,3H),3.75(s,3H),1.68(tt,J=8.2,4.6Hz,1H),1.03–0.95(m,2H),0.77(dq,J=7.0,3.4Hz,2H). LC-MS(ESI)m / z 538[M+H]

[0567] I-6

[0568]

[0569] The I-6 was manufactured in a similar manner to the I-5

[0570] 1H NMR (400MHz, DMSO-d6) δ8.70–8.64(m,1H),8.17(d,J=8.0Hz,1H),7.53(q,J=8.4Hz,4H),6.76(s,1H),4.98(d,J=3.6Hz,2H ), 4.36 (s, 2H), 3.91–3.86 (m, 3H), 3.72 (s, 2H), 2.35–2.30 (m, 3H), 1.87 (s, 1H), 1.02 (s, 2H), 0.85 (dd, J = 8.0, 3.4Hz, 2H). LC-MS(ESI)m / z 566[M+H].

[0571] I-7

[0572]

[0573] 1-7 was synthesized in a similar manner to 1-2.

[0574] 1 H NMR (400MHz, DMSO-d6) δ8.70–8.64(m,1H),8.17(d,J=8.0Hz,1H),7.53(q,J=8.4Hz,4H),6.76(s,1H),4.98(d,J=3.6Hz,2H ), 4.36 (s, 2H), 3.91–3.86 (m, 3H), 3.72 (s, 2H), 2.35–2.30 (m, 3H), 1.87 (s, 1H), 1.02 (s, 2H), 0.85 (dd, J = 8.0, 3.4Hz, 2H). LC-MS(ESI)m / z 524[M+H]

[0575] I-8

[0576]

[0577] Step 1 :2-chloro-8-({4-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one

[0578] To a stirred solution of 2-chloro-8H-pyrido[2,3-d]pyrimidin-7-one (50 mg, 0.275 mmol, 1 eq) and 1-[4-(chloromethyl)phenyl]-5-methyl-3-(trifluoromethyl)pyrazole (68.07 mg, 0.248 mmol, 0.9 eq) in DMF (1 mL) was added 1,1,3,3-tetramethylguanidine (47.57 mg, 0.413 mmol, 1.5 eq) at room temperature. The resulting mixture was stirred at room temperature under a nitrogen atmosphere overnight. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% TFA) over 10 min with UV detector (254 nm) to afford 2-chloro-8-({4-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one (60 mg, 51.91%) as a white solid. LC-MS (ESI) m / z 420.7 [M+H].

[0579] Step 2: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one

[0580] To a stirred solution of 2-chloro-8-({4-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one (50 mg, 0.119 mmol, 1 eq) and 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (34.66 mg, 0.178 mmol, 1.5 eq) in 1,4-dioxane (1 mL) and H2O (0.2 mL) was added K3PO4 (63.21 mg, 0.297 mmol, 2.5 eq) and Pd(dppf)Cl2 (17.43 mg, 0.024 mmol, 0.2 eq) at room temperature. The resulting mixture was stirred at 90 ° C under a nitrogen atmosphere for 2 hours. The resulting mixture was filtered and the filter cake was washed with 1,4-dioxane (3 x 5 mL). The filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC under the following conditions (column: Xselect CSH C18 OBD column 30*150mm 5μm, n; mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 55% B to 95% B, 95% B in 7 min; wavelength: 254 / 220 nm; RT1 (min): 6.02) to give 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one (15.0 mg, 22.68%) as an off-white solid. 1 H NMR (300MHz, DMSO-d6) δ9.30(s,1H),8.70(s,1H),8.17(d,J=9.5Hz,1H),7.68–7.27(m,4H),6.92(d,J=9.5Hz,1H),6.74(s, 1H), 5.59 (s, 2H), 3.83 (s, 3H), 2.31 (d, J = 0.7Hz, 3H), 1.81–1.65 (m, 1H), 1.02 (d, J = 4.2Hz, 2H), 0.78 (dd, J = 7.9, 3.3Hz, 2H). LC-MS (ESI) m / z 534.1 [M+H].

[0581] I-9

[0582]

[0583] Step 1: Ethyl 2-[(6-chloropyridin-3-yl)oxy]acetate

[0584] A solution of 6-chloropyridin-3-ol (7.7 g, 59.441 mmol, 1 eq) in ACN (154.01 mL) was treated with potassium carbonate (23.17 g, 166.435 mmol, 2.8 eq) and ethyl chloroacetate (9.11 g, 74.301 mmol, 1.25 eq) at 60 ° C under a nitrogen atmosphere for 4 h. The mixture was allowed to cool to room temperature. The resulting mixture was filtered and concentrated under reduced pressure to give ethyl 2-[(6-chloropyridin-3-yl)oxy]acetate (8 g, 62.42%) as a reddish-brown solid. LC-MS (ESI) m / z 216.0 [M + H]

[0585] Step 2 : ethyl 2-[(6-chloro-1-oxo-1λ5-pyridin-3-yl)oxy]acetate

[0586] Under nitrogen atmosphere at 50 DEG C by 2- [(6- chloropyridin-3-yl) oxy] ethyl acetate (5g, 23.188mmol, 1 equivalent) in DCE (100.00mL, 1263.282mmol, 54.48 equivalent) solution with m-CPBA (8.00g, 46.376mmol, 2.0 equivalent) treatment 4h, and then stirred at room temperature overnight. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% ACN / water (0.1% FA) in 10min with UV detector (254nm). The resulting mixture was concentrated under reduced pressure and freeze-dried to give 2- [(6- chloro-1-oxo-1λ5-pyridin-3-yl) oxy] ethyl acetate (4g, 74.47%) as a yellow solid. LC-MS (ESI) m / z 232.15 [M+H]

[0587] Step 3 Methyl 2-[(6-chloro-4-nitro-1-oxo-1λ5-pyridin-3-yl)oxy]acetate

[0588] A solution of ethyl 2-[(6-chloro-1-oxo-1λ5-pyridin-3-yl)oxy]acetate (1000 mg, 4.317 mmol, 1 eq) in H2SO4 (4 mL, 75.048 mmol, 17.38 eq) was treated dropwise with HNO3 (2 mL, 44.595 mmol, 10.33 eq) at 0°C under a nitrogen atmosphere for several minutes. The reaction mixture was heated to 40°C in an oil bath. The temperature was slowly raised to 75°C over 1 h and then maintained for 2 h. The mixture was then ice-cooled, adjusted to pH 9 by adding 50% NaOH, dried in vacuo, washed with 3 x 2 mL MeOH, and concentrated under reduced pressure. The residue was dissolved in MeOH (50 ml) and treated with H2SO4 (1 mL) at 70°C under a nitrogen atmosphere for 2 h. The resulting mixture was concentrated under reduced pressure, diluted with water (20 ml), basified to pH 9 with NaOH, and extracted with EtOAc (3 x 15 mL). The combined organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% ACN / water (0.1% FA) in 10 min with a UV detector (254 nm), concentrated under reduced pressure, and freeze-dried to obtain 2- [(6- chloro-4-nitro-1-oxo-1λ5-pyridin-3-yl) oxy] methyl acetate (200 mg, 17.64%) as a white solid. LC-MS (ESI) m / z 263.15 [M+H]

[0589] Step 4: 7-Chloro-1H,3H-pyrido[3,4-b][1,4]oxazin-2-one

[0590] A solution of methyl 2-[(6-chloro-4-nitro-1-oxo-1λ5-pyridin-3-yl)oxy]acetate (210 mg, 0.800 mmol, 1 eq) in MeOH (6 mL) was treated with Fe (312.61 mg, 5.600 mmol, 7 eq) and AcOH (0.25 mg, 0.004 mmol, 0.01 eq) at 70 ° C under a nitrogen atmosphere for 4 h. The hot solution was filtered through a pad of celite and concentrated. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% ACN / water (0.1% NH 3 -H 2 O) over 10 min with a UV detector (254 nm) to produce 7-chloro-1H,3H-pyrido[3,4-b][1,4]oxazin-2-one (140 mg, 94.85%) as a reddish-brown solid. LC-MS (ESI) m / z 185.00 [M+H]

[0591] Step 57-Chloro-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-3H-pyrido[3,4-b][1,4]oxazin-2-one

[0592] A solution of 7-chloro-1H,3H-pyrido[3,4-b][1,4]oxazin-2-one (200 mg, 1.084 mmol, 1 eq) in DMF (10.00 mL) was treated with KCO (449.25 mg, 3.252 mmol, 3 eq) and 2-[4-(bromomethyl)phenyl]-1-methyl-4-(trifluoromethyl)imidazole (449.52 mg, 1.409 mmol, 1.3 eq) at 50° C. under nitrogen atmosphere for 3 h. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% ACN / water (0.1% NH3-H2O) over 10 min with UV detector (254 nm) to afford 7-chloro-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-3H-pyrido[3,4-b][1,4]oxazin-2-one (250 mg, 54.57%) as an off-white solid. LC-MS (ESI) m / z 423.2 [M+H]

[0593] Step 6. 2-[4-({7-chloro-2H,3H-pyrido[3,4-b][1,4]oxazin-1-yl}methyl)phenyl]-1-methyl-4-(trifluoromethyl)imidazole

[0594] A mixture of 7-chloro-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-3H-pyrido[3,4-b][1,4]oxazin-2-one (100 mg, 0.237 mmol, 1 eq) and BH3-THF (0.59 mL, 0.593 mmol, 2.5 eq, 1 M THF solution) in THF (1.5 mL) was stirred at 75 ° C under a nitrogen atmosphere for 1 h. The reaction was quenched with MeOH at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% ACN / water (0.1% FA) over 10 min with UV detector (254 nm)) and freeze-dried to give 2-[4-({7-chloro-2H,3H-pyrido[3,4-b][1,4]oxazin-1-yl}methyl)phenyl]-1-methyl-4-(trifluoromethyl)imidazole (60 mg, 62.05%) as an off-white solid. LC-MS (ESI) m / z 409.1 [M+H]

[0595] Step 7.4-Cyclopropyl-6-methoxy-5-[1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-2H,3H-pyrido[3,4-b][1,4]oxazin-7-yl]pyrimidine

[0596] A solution of 2-[4-({7-chloro-2H,3H-pyrido[3,4-b][1,4]oxazin-1-yl}methyl)phenyl]-1-methyl-4-(trifluoromethyl)imidazole (60 mg, 0.147 mmol, 1 eq) and 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (42.71 mg, 0.220 mmol, 1.5 eq) in 1,4-dioxane (1.8 mL) and H2O (0.2 mL) was treated with K3PO4 (77.88 mg, 0.367 mmol, 2.5 eq) and Pd(dppf)Cl2 (32.22 mg, 0.044 mmol, 0.3 eq) at 90°C under nitrogen atmosphere for 2 h. The residue was purified by reverse phase flash chromatography (C18 silica column, eluting with 0% to 100% ACN / water (0.1% NH3-H2O) over 10 min with UV detection (254 nm). The crude product was purified by preparative HPLC under the following conditions (column: Sunfireprep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 15% B to 45% B, 45% B in 7 min; wavelength: 254 / 220 nm; RT1 (min): 4.45) to give 4-cyclopropyl-6-methoxy-5-[1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-2H,3H-pyrido[3,4-b][1,4]oxazin-7-yl]pyrimidine (10.9 mg, 14.21%) as an off-white solid. 1 H-NMR(400MHz,DMSO-d6):1H NMR (400MHz, DMSO-d6) δ8.54(s,1H),7.92(d,J=4.5Hz,2H),7.70(d,J=7.9Hz,2H),7.42(d,J=7.9Hz,2H),6.80(s,1H),4.68(s,2H),4.31(t, J=4.3Hz,2H),3.75(d,J=11.0Hz,6H),3.57(t,J=4.4Hz,2H),1.78(dd,J=8.2,3.9Hz,1H),0.93(p,J=3.7Hz,2H),0.77(dq,J=7.1,3.4Hz,2H). LC-MS (ESI) m / z 523.25 [M+H]

[0597] I-10

[0598]

[0599] A solution of 7-chloro-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-3H-pyrido[3,4-b][1,4]oxazin-2-one (80 mg, 0.189 mmol, 1 eq) and 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (55.06 mg, 0.283 mmol, 1.5 eq) in 1,4-dioxane (1.8 mL) and H2O (0.16 mL, 8.872 mmol, 46.94 eq) was treated with K3PO4 (100.41 mg, 0.473 mmol, 2.5 eq) and Pd(dppf)Cl2 (27.69 mg, 0.038 mmol, 0.2 eq) at 90°C under nitrogen atmosphere for 2 h. The resulting mixture was filtered and the filter cake was washed with 1,4-dioxane (3x10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC under the following conditions (column: XBridge PrepOBD C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.05% NH3-H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 28% B to 60% B, 60% B in 8 min; wavelength: 220 nm; RT1 (min): 7.35) to obtain 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-3H-pyrido[3,4-b][1,4]oxazin-2-one (20.4 mg, 20.05%) as an off-white solid. 1 H-NMR (400MHz, DMSO-d6): δ8.59(s,1H),8.35(s,1H),7.92(d,J=1.3Hz,1H),7.73–7.66(m,2H),7.45(d,J=8.1Hz,2H),7.29(s,1H), 5.26(s,2H),5.01(s,2H),3.76(s,3H),3.69(s,3H),1.71(tt,J=8.3,4.6Hz,1H),0.95(p,J=3.7Hz,2H),0.76(dq,J=6.9,3.5Hz,2H). LC-MS(ESI)m / z 537.2[M+H].

[0600] I-11

[0601]

[0602] Step 1:1-{6-chloro-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridin-3-yl}-2-methoxyethanone

[0603] To a stirred solution of ethyl 4,6-dichloropyridine-3-carboxylate (100 mg, 0.454 mmol, 1 eq) and 1-{4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methanamine (127.59 mg, 0.499 mmol, 1.1 eq) in tetrahydrofuran (2 mL, 20 eq) was added triethylamine (91.7 mg, 0.908 mmol, 2 eq) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 60 ° C. under a nitrogen atmosphere for 1 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give 1-{6-chloro-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridin-3-yl}-2-methoxyethanone as a light yellow solid (78 mg, 39.11%). LC-MS (ESI) m / z 439.1 [M+H]

[0604] Step 2. {6-chloro-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridin-3-yl}methanol

[0605] To a stirred solution of 1-{6-chloro-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridin-3-yl}-2-methoxyethanone (78 mg, 0.178 mmol, 1 eq) in tetrahydrofuran (1.5 mL, 20 eq) was added LiAlH4 (20.24 mg, 0.534 mmol, 3 eq) in portions at 0 ° C under a nitrogen atmosphere. The resulting mixture was stirred at 0 ° C for 1 h under a nitrogen atmosphere. The reaction was quenched with 10H2O.Na2SO4 (100 mg) at 0 ° C. The resulting mixture was filtered and the filter cake was washed with THF (3x5 mL). The filtrate was concentrated under reduced pressure. The residue was dissolved in THF (2 mL). The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 10% to 50% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give {6-chloro-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridin-3-yl}methanol (23.5 mg, 33.32%) as a white solid. LC-MS (ESI) m / z 397.1 [M+H]

[0606] Step 3. 6-Chloro-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridine-3-carbaldehyde

[0607] To a stirred solution of {6-chloro-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridin-3-yl}methanol (85 mg, 0.214 mmol, 1 eq) in dichloromethane (4 mL, 20 eq) was added manganese dioxide (55.87 mg, 0.642 mmol, 3.0 eq) at room temperature under a nitrogen atmosphere. The reaction mixture was kept at room temperature overnight. The resulting mixture was filtered and the filter cake was washed with CHCl (3 x 5 mL). The filtrate was concentrated under reduced pressure. The residue was dissolved in MeOH (2 mL). The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detection (254 nm) to give 6-chloro-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridine-3-carbaldehyde (60 mg, 70.95%) as a pale yellow solid. LC-MS (ESI) m / z 395.0 [M+H].

[0608] Step 4. 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridine-3-carbaldehyde

[0609] To a stirred mixture of 6-chloro-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridine-3-carbaldehyde (20 mg, 0.051 mmol, 1 eq) and 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (14.74 mg, 0.076 mmol, 1.5 eq) in 1,4-dioxane (0.9 mL, 1.020 mmol, 20 eq) and water (0.1 mL, 1.020 mmol, 20 eq) was added K 3 PO 4 (32.26 mg, 0.153 mmol, 3 eq) and Pd(dppf)Cl 2 (7.41 mg, 0.010 mmol, 0.2 eq) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 100 ° C. for 2 h under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in THF (1 mL). The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to afford 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridine-3-carbaldehyde (15 mg, 58.23%) as a yellow solid. LC-MS (ESI) m / z 509.1 [M+H]

[0610] Step 5: 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-1,6-naphthyridin-2-one

[0611] To a stirred solution of ethyl acetate (34.65 mg, 0.392 mmol, 4.0 equiv) in THF (2 mL, 20 equiv) was added LiHMDS (0.392 mL, 0.392 mmol, 4.0 equiv, 1 M THF solution) dropwise at -78 ° C under a nitrogen atmosphere. The resulting mixture was stirred at -78 ° C for 10 min under a nitrogen atmosphere. 6- (4-cyclopropyl-6-methoxypyrimidin-5-yl) -4- [( {4- [1-methyl-4- (trifluoromethyl) imidazole-2-yl] phenyl} methyl) amino] pyridine-3-carbaldehyde (50 mg, 0.098 mmol, 1 equiv) was added dropwise at -78 ° C. The resulting mixture was stirred at room temperature for another 3 h. The reaction was quenched with water at 0 ° C. The resulting mixture was extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (2 x 2 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (XBridge Shield RP18 OBD column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 40% B to 75% B, 75% B in 7 min; wavelength: 254 / 220 nm; RT1 (min): 3.45;) to give 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-1,6-naphthyridin-2-one (21.1 mg, 40.26%) as a white solid. 1 H NMR (300MHz, DMSO-d6) δ9.03(s,1H),8.64(s,1H),8.17(d,J=9.6Hz,1H),7.92(d,J=1.4Hz,1H),7.73–7.61(m,3H),7.37(d,J=8.1Hz,2H ), 6.88(d,J=9.5Hz,1H),5.58(s,2H),3.75(s,3H),3.69(s,3H),1.74–1.64(m,1H),0.97(d,J=4.4Hz,2H),0.75(dd,J=8.1,3.3Hz,2H). LC-MS(ESI)m / z 533.10[M+H]

[0612] I-89

[0613]

[0614] Step 1: 2-Chloro-N-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5-nitropyridin-4-amine

[0615] A mixture of 2,4-dichloro-5-nitropyridine (200 mg, 1.036 mmol, 1 equivalent), 1-{4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methylamine (310 mg, 1.215 mmol, 1.17 equivalents) and TEA (65 mg, 1.5 mmol, 1.2 equivalents) in THF (10 mL) was stirred for 2 h at room temperature under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% MeCN / water (0.1% FA) in 10 min with a UV detector (254 nm) to give 2-chloro-N-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5-nitropyridine-4-amine (260 mg, 60.93%) as a colorless oil. LC-MS (ESI) m / z 412 [M+H].

[0616] Step 2: 6-Chloro-N4-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyridine-3,4-diamine

[0617] A solution of 2-chloro-N-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5-nitropyridin-4-amine (260 mg, 0.631 mmol, 1 eq), iron (280 mg, 5.014 mmol, 7.94 eq), NHCl (180 mg, 3.365 mmol, 5.33 eq) and H0 (2 mL, 111.019 mmol, 175.82 eq) in EtOH (8 mL) was stirred under nitrogen for 2 h at 40° C. The resulting mixture was filtered and the filter cake was washed with EtOH (2×100 mL). The filtrate was concentrated under reduced pressure to give 6-chloro-N4-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyridine-3,4-diamine (350 mg, 145.19%) as a brown solid. LC-MS (ESI) m / z 382 [M+H].

[0618] Step 3: N-(5-amino-2-chloropyridin-4-yl)-2-chloro-N-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)acetamide

[0619] To a solution of 6-chloro-N4-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyridine-3,4-diamine (450 mg, 1.179 mmol, 1 eq) in DCM (10 mL) was added TEA (430 mg, 4.249 mmol, 3.61 eq) at room temperature under a nitrogen atmosphere, followed by the addition of chloroacetyl chloride (170 mg, 1.505 mmol, 1.28 eq) dropwise at 0°C. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to afford N-(5-amino-2-chloropyridin-4-yl)-2-chloro-N-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)acetamide (150 mg, 27.77%) as a colorless oil. LC-MS (ESI) m / z 458 [M+H].

[0620] Step 4: 7-Chloro-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-2H,4H-pyrido[3,4-b]pyrazin-3-one

[0621] A solution of 2-chloro-N-{6-chloro-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridin-3-yl}acetamide (100 mg, 0.218 mmol, 1 eq) and KCO (60 mg, 0.434 mmol, 1.99 eq) in DMF (5 mL) was stirred under nitrogen at 80° C. for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to afford 7-chloro-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-2H,4H-pyrido[3,4-b]pyrazin-3-one (30 mg, 32.59%) as an off-white solid. LC-MS (ESI) m / z 422 [M+H].

[0622] Step 5. Synthesis of 4-cyclopropyl-5-[1-({4-[1-ethyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrazolo[4,3-c]pyridin-6-yl]-6-methoxypyrimidine

[0623] A solution of 2-[4-({6-chloropyrazolo[4,3-c]pyridin-1-yl}methyl)phenyl]-1-ethyl-4-(trifluoromethyl)imidazole (70 mg, 0.172 mmol, 1 eq), 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (38 mg, 0.196 mmol, 1.14 eq), and Pd(dppf)Cl (20 mg, 0.027 mmol, 0.16 eq) in H O (1 mL) and dioxane (4 mL) was stirred at 90° C. under a nitrogen atmosphere for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give 4-cyclopropyl-5-[1-({4-[1-ethyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrazolo[4,3-c]pyridin-6-yl]-6-methoxypyrimidine as an off-white solid (8 mg, 8.63%) 1 H-NMR:1H NMR (400MHz, DMSO-d6) δ10.75(s,1H),8.55(s,1H),7.94(d,J=8.8Hz,2H),7.70(d,J=8.0Hz,2H),7.45(d,J=7.9Hz,2H),6. 82(s,1H),4.63(s,2H),4.05(s,2H),3.76(d,J=10.6Hz,6H),1.80(s,1H),1.24(s,1H),0.94(s,2H),0.79(d,J=7.7Hz,2H). LC-MS(ESI)m / z536.15[M+H].

[0624] I-13

[0625]

[0626] Step 1: 2-Chloro-N-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5-nitropyridin-4-amine

[0627] To a stirred solution of 2,4-dichloro-5-nitropyridine (415.83 mg, 2.155 mmol, 1.1 eq) and TEA (396.46 mg, 3.918 mmol, 2 eq) in THF (4 mL) was added a solution of 1-{4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methanamine (500 mg, 1.959 mmol, 1.00 eq) in THF (1 mL) dropwise at 0° C. The resulting mixture was stirred at room temperature for 2 h and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 10% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to afford 2-chloro-N-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5-nitropyridin-4-amine (400 mg, 49.5%) as a yellow solid. LC-MS (ESI) m / z 412.7 [M+H].

[0628] Step 2: 6-Chloro-N4-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyridine-3,4-diamine

[0629] To a stirred solution of 2-chloro-N-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5-nitropyridin-4-amine (150 mg, 0.364 mmol, 1 eq) and NHCl (97.43 mg, 1.820 mmol, 5 eq) in ethanol (2.5 mL) and water (0.6 mL) was added Fe (122.06 mg, 2.184 mmol, 6 eq) at room temperature. The resulting mixture was stirred at 40 ° C for 2 h. The resulting mixture was filtered and the filter cake was washed with ethanol (3 x 10 mL). The filtrate was concentrated under reduced pressure. The crude product was used directly in the next step without further purification. LC-MS (ESI) m / z 382.7 [M+H].

[0630] Step 3: 7-Chloro-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[3,4-b]pyrazin-2-one

[0631] To a stirred solution of 6-chloro-N4-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyridine-3,4-diamine (130 mg, 0.341 mmol, 1 eq) and ethyl glyoxylate (104.28 mg, 1.023 mmol, 3 eq) in MeOH (2 mL) was added AcOH (0.1 mL) dropwise at room temperature. The resulting mixture was stirred at 100 ° C for 2 h and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 10% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to afford 7-chloro-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[3,4-b]pyrazin-2-one (60 mg, 41.9%) as a tan solid. LC-MS (ESI) m / z 420.7 [M+H].

[0632] Step 4: 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[3,4-b]pyrazin-2-one

[0633] To a stirred solution of 7-chloro-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[3,4-b]pyrazin-2-one (60 mg, 0.143 mmol, 1 eq) and 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (41.59 mg, 0.214 mmol, 1.5 eq) in 1,4-dioxane (2 mL) and H2O (0.4 mL) was added Pd(dppf)Cl2 (20.92 mg, 0.029 mmol, 0.2 eq) and K3PO4 (75.85 mg, 0.357 mmol, 2.50 eq) at room temperature. The resulting mixture was stirred at 90 ° C for 2 h. The resulting mixture was filtered and the filter cake was washed with 1,4-dioxane (2 x 10 mL). The filtrate was concentrated under reduced pressure. The crude product was purified by reverse phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN in water (0.1% FA) over 10 min with UV detector (254 nm) to give 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[3,4-b]pyrazin-2-one (7.7 mg, 9.88%) as a tan solid. 1H NMR (300MHz, DMSO-d6) δ9.12(s,1H),8.65(s,1H),8.46(s,1H),7.93(s,1H),7.75(s,1H),7.69(d,J=8.1Hz,2H),7.47(d, J=8.1Hz,2H),5.52(s,2H),3.73(d,J=12.9Hz,6H),1.81–1.62(m,1H),0.99(d,J=4.1Hz,2H),0.78(dd,J=8.1,3.3Hz,2H). LC-MS(ESI)m / z 534.2[M+H].

[0634] I-14

[0635]

[0636] The I-14 is manufactured in a similar manner to the I-2 1 H NMR (400MHz, DMSO-d6) δ8.67(d,J=2.0Hz,1H),8.18(s,1H),8.03(d,J=1.5Hz,1H),7.61(d,J=8.1Hz,2H),7.47(d,J=8.0Hz,2H),4.97(s,2H),4.36(s, 2H),4.07(q,J=7.3Hz,2H),3.89(s,3H),3.71(s,2H),1.87(d,J=7.3Hz,1H) ,1.32(t,J=7.2Hz,3H), 1.01(d,J=4.4Hz,2H), 0.84(dt,J=8.0,3.4Hz,2H). LC-MS(ESI)m / z 538[M+H].

[0637] I-15

[0638]

[0639] The I-15 was manufactured in a similar manner to the I-4. 1H NMR (400MHz, DMSO-d6) δ9.29(s,1H),8.69(s,1H),8.16(d,J=9.5Hz,1H),8.00(d,J=1.3Hz,1H),7.57–7.50(m,2H),7.40(d,J=8.3Hz,2H ), 6.91 (d, J = 9.5Hz, 1H), 5.57 (s, 2H), 4.04 (q, J = 7.2Hz, 2H), 3.82 (s, 3H), 1.72 (m, 1H), 1.29 (t, J = 7.2Hz, 3H), 1.01 (m, 2H), 0.76 (m, 2H). LC-MS(ESI)m / z:548.0[M+H]

[0640] I-16

[0641]

[0642] Step 1: 1-Methyl-4-(trifluoromethyl)imidazole

[0643] Under nitrogen atmosphere, 4-(trifluoromethyl)-1H-imidazole (3g, 22.046mmol, 1 equivalent) in THF (30mL) was treated with NaH (0.79g, 33.069mmol, 1.5 equivalents) for 30min at 0°C, and then MeI (3.13g, 22.046mmol, 1 equivalent) was added dropwise at 0°C. The resulting mixture was stirred for 2h at room temperature under nitrogen atmosphere. The mixture was quenched with saturated NH4Cl (aqueous solution) (10mL). The resulting mixture was extracted with EtOAc (3x 100mL). The combined organic layer was washed with water (2x50mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN in water (0.1% FA) over 10 min with UV detector (254 nm) to give 1-methyl-4-(trifluoromethyl)imidazole (1.5 g, 45.33%) as a colorless oil. LC-MS (ESI) m / z 150 [M+H].

[0644] Step 2: 2-Bromo-1-methyl-4-(trifluoromethyl)imidazole

[0645] Under nitrogen atmosphere at -78 DEG C by 1- methyl -4- (trifluoromethyl) imidazole (100mg, 0.666mmol, 1 equivalent) in THF (10mL) solution with n-butyl lithium (76.82mg, 1.199mmol, 1.8 equivalent) treatment 30min, then at -78 DEG C add CBr4 (441.86mg, 1.332mmol, 2 equivalents). The resulting mixture was stirred at room temperature under nitrogen atmosphere for 2h. The mixture was acidified to pH 5 with 1M HCl (5mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% MeCN / water (0.1% FA) in 10min with UV detector (254nm)) to obtain 2- bromo-1-methyl -4- (trifluoromethyl) imidazole (50mg, 32.77%) as a colorless oil. LC-MS (ESI) m / z 229 [M+H].

[0646] Step 3: {1-[1-Methyl-4-(trifluoromethyl)imidazol-2-yl]piperidin-4-yl}methanol

[0647] To a stirred solution of 2-bromo-1-methyl-4-(trifluoromethyl)imidazole (50 mg, 0.233 mmol, 1 eq) and piperidin-4-ylmethanol (80.37 mg, 0.699 mmol, 3 eq) in n-BuOH (2 mL, 21.880 mmol, 94.07 eq) was added DIEA (90.18 mg, 0.699 mmol, 3 eq) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 160 ° C. overnight under a nitrogen atmosphere. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% MeCN / water (0.1% FA) over 10 min with a UV detector (254 nm) to give {1-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]piperidin-4-yl}methanol (50 mg, 81.66%) as a yellow oil. LC-MS (ESI) m / z 263 [M+H].

[0648] Step 4: 4-(Chloromethyl)-1-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]piperidine

[0649] To a stirred solution of {1-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]piperidin-4-yl}methanol (180 mg, 0.684 mmol, 1 equiv), pyridine (0.1 mL, 0.001 mmol, 0.01 equiv) in DCM (3 mL) was added thionyl chloride (0.5 mL, 0.004 mmol, 0.01 equiv) at 0 ° C under a nitrogen atmosphere. The resulting mixture was stirred overnight at 40 ° C under a nitrogen atmosphere. The reaction was quenched with water (20 mL) at room temperature. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% MeCN / water (0.1% FA) in 10 min with a UV detector (254 nm) to obtain 4-(chloromethyl)-1-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]piperidine (85 mg, 44.13%) as a yellow solid. LC-MS (ESI) m / z 282 [M+H].

[0650] Step 5: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({1-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]piperidin-4-yl}methyl)pyrido[2,3-d]pyrimidin-7-one

[0651] A solution of 4-(chloromethyl)-1-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]piperidine (70 mg, 0.248 mmol, 1 eq), 1,1,3,3-tetramethylguanidine (42.93 mg, 0.372 mmol, 1.5 eq) and 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8H-pyrido[2,3-d]pyrimidin-7-one (73.38 mg, 0.248 mmol, 1 eq) in DMF (27.24 mg, 0.372 mmol, 1.5 eq) was stirred overnight at 100° C. under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN in water (0.1% FA) over 10 min with UV detector (254 nm) to give 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({1-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]piperidin-4-yl}methyl)pyrido[2,3-d]pyrimidin-7-one (12.3 mg, 9.08%) as an off-white solid. 1 H-NMR: 1H NMR(400MHz,DMSO-d6)δ9.24(s,1H),8.71(s,1H),8.10(d,J=9.5Hz,1H),7.51(s,1H),6 .85(d,J=9.5Hz,1H),4.29(d,J=7.2Hz,2H),3.87(s,3H),3.46(s,3H),3.22(d,J=12.2H z,3H),2.62(t,J=11.8Hz,2H),2.06(d,J=16.2Hz,1H),1.80(td,J=8.3,4.4Hz,1H),1.5 9(d,J=12.7Hz,2H),1.53–1.36(m,2H),1.13–1.07(m,2H),0.91(dq,J=7.0,3.4Hz,2H). LC-MS(ESI)m / z 541.10[M+H].

[0652] I-17

[0653]

[0654] Step 1: 2-Chloro-N-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5-nitropyrimidin-4-amine

[0655] To a stirred solution of 2,4-dichloro-5-nitropyrimidine (400 mg, 2.062 mmol, 1 eq) and TEA (626.03 mg, 6.186 mmol, 3 eq) in THF (8 mL) was added dropwise a solution of (4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanamine (547.17 mg, 2.268 mmol, 1.1 eq) in THF (1 mL) at 0 ° C. The resulting mixture was stirred at room temperature for 2 h. The reaction was quenched by adding water (20 mL) at room temperature. The resulting mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with water (3 x 50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to afford 2-chloro-N-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5-nitropyrimidin-4-amine (160 mg, 18.80%) as a pale yellow oil. LC-MS (ESI) m / z 413.7 [M+H].

[0656] Step 2: 2-Chloro-N4-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrimidine-4,5-diamine

[0657] To a stirred solution of 2-chloro-N-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5-nitropyrimidin-4-amine (50 mg, 0.121 mmol, 1 eq) and NHCl (32.40 mg, 0.605 mmol, 5 eq) in ethanol (0.8 mL) and water (0.2 mL) was added Fe (54.12 mg, 0.968 mmol, 8 eq) at room temperature. The resulting mixture was stirred at 40 °C for 2 h. The resulting mixture was filtered and the filter cake was washed with ethanol (3 x 10 mL). The filtrate was concentrated under reduced pressure. The crude product was used in the next step without further purification. LC-MS (ESI) m / z 383.7 [M+H].

[0658] Step 3: 7-Chloro-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[3,4-b]pyrazin-2-one

[0659] To a stirred solution of 2-chloro-N4-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrimidine-4,5-diamine (150 mg, 0.392 mmol, 1 eq) and ethyl glyoxylate (120.02 mg, 1.176 mmol, 3 eq) in MeOH (2 mL) was added AcOH (20.45 mg, 0.341 mmol, 1 eq) dropwise at room temperature. The resulting mixture was stirred at 100 ° C for 2 h and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to afford 7-chloro-1-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[3,4-b]pyrazin-2-one (40 mg, 24.3%) as a yellow oil. LC-MS (ESI) m / z 420.7 [M+H].

[0660] Step 4 :2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pteridin-7-one

[0661] To a stirred solution of 2-chloro-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pteridin-7-one (50 mg, 0.119 mmol, 1 eq) and 4-cyclopropyl-6-hydroxypyrimidin-5-ylboronic acid (32.08 mg, 0.178 mmol, 1.5 eq) in 1,4-dioxane (0.4 mL) and H2O (0.1 mL) was added K3PO4 (63.06 mg, 0.297 mmol, 2.5 eq) and Pd(dppf)Cl2 (17.39 mg, 0.024 mmol, 0.2 eq) at room temperature. The resulting mixture was stirred at 90°C under a nitrogen atmosphere for 2 hours. The resulting mixture was filtered; the filter cake was washed with 1,4-dioxane (3 x 5 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica column, eluting with 0% to 100% MeCN in water (0.1% FA) over 10 min with UV detector (254 nm) to give 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pteridin-7-one (16.1 mg, 23.35%) as a light yellow solid. 1H NMR(400MHz, DMSO-d6)δ9.37(d,J=1.5Hz,1H),8.70(s,1H),8.48(s,1H),7.92(s,1H),7.69–7.57(m,2H), 7.48(d,J=8.0Hz,2H),5.49(s,2H),3.79(dd,J=40.2,1.4Hz,6H),1.76(m,1H),1.03(m,2H),0.78(m,2H). LC-MS(ESI)m / z 535.1[M+H].

[0662] I-18

[0663]

[0664] Step 1 2-Chloro-N-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5-nitropyrimidin-4-amine

[0665] A solution of 2,4-dichloro-5-nitropyrimidine (200 mg, 1.031 mmol, 1 eq), TEA (310 mg, 3.063 mmol, 2.97 eq) and 1-{4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methanamine (310 mg, 1.215 mmol, 1.18 eq) in THF (10 mL) was stirred at room temperature under a nitrogen atmosphere for 2 h. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with a UV detector (254 nm) to give 2-chloro-N-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5-nitropyrimidin-4-amine (336 mg, 78.95%) as a colorless oil. LC-MS (ESI) m / z 413 [M+H].

[0666] Step 2 :2-chloro-N4-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrimidine-4,5-diamine

[0667] A solution of 2-chloro-N-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5-nitropyrimidin-4-amine (375 mg, 0.909 mmol, 1 eq), iron (400 mg, 7.163 mmol, 7.88 eq), NHCl (240 mg, 4.487 mmol, 4.94 eq) and H0 (1 mL, 55.509 mmol, 61.10 eq) in EtOH (4 mL) was stirred at 40 °C under a nitrogen atmosphere for 2 h. The resulting mixture was filtered and the filter cake was washed with MeOH (2 x 10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE / EA (1:1)) to give 2-chloro-N4-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrimidine-4,5-diamine (175 mg, 50.32%) as a yellow solid. LC-MS (ESI) m / z 383 [M+H].

[0668] Step 3: 2-Chloro-N-{2-chloro-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyrimidin-5-yl}acetamide

[0669] A solution of 2-chloro-N4-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrimidine-4,5-diamine (200 mg, 0.522 mmol, 1 eq) in DCM (2 mL, 31.461 mmol, 60.21 eq) was treated with TEA (1.4 mL, 10.072 mmol, 19.28 eq) at 0° C. under a nitrogen atmosphere for 5 min, followed by the dropwise addition of chloroacetyl chloride (150 mg, 1.328 mmol, 2.54 eq) at 0° C. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give 2-chloro-N-{2-chloro-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyrimidin-5-yl}acetamide (100 mg, 41.67%) as a colorless oil. LC-MS (ESI) m / z 459 [M+H].

[0670] Step 4: 2-Chloro-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5,7-dihydropteridin-6-one

[0671] A solution of 2-chloro-N-{2-chloro-4-[({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyrimidin-5-yl}acetamide (130 mg, 0.283 mmol, 1 eq) and KCO (78.24 mg, 0.566 mmol, 2 eq) in DMF (10 mL, 129.215 mmol, 456.48 eq) was stirred under nitrogen for 2 h at 80° C. The mixture was allowed to cool to room temperature. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to afford 2-chloro-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5,7-dihydropteridin-6-one (23 mg, 19.22%) as an off-white solid. LC-MS (ESI) m / z 423 [M+H].

[0672] Step 5: 2-Chloro-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-6,7-dihydro-5H-pteridine

[0673] A solution of 2-chloro-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-5,7-dihydropteridin-6-one (20 mg, 0.047 mmol, 1 eq) and BH3-THF (1.4 mL, 14.629 mmol, 309.25 eq) in THF (2 mL, 24.686 mmol, 521.85 eq) was stirred at 65° C. under a nitrogen atmosphere for 2 h. The reaction was quenched by the addition of MeOH (4 mL) at 0° C. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give 2-chloro-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-6,7-dihydro-5H-pteridine (10 mg, 51.71%) as a colorless solid. LC-MS (ESI) m / z 409 [M+H].

[0674] Step 6: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-6,7-dihydro-5H-pteridine

[0675] A solution of 2-chloro-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-6,7-dihydro-5H-pteridine (50 mg, 0.122 mmol, 1 eq), 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (28.47 mg, 0.146 mmol, 1.2 eq), Pd(dppf)Cl (17.90 mg, 0.024 mmol, 0.2 eq), H O (1 mL, 55.509 mmol, 453.86 eq), and K PO (38.94 mg, 0.183 mmol, 1.5 eq) in dioxane (4 mL) was stirred under a nitrogen atmosphere for 2 h at 90° C. The mixture was allowed to cool to room temperature. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to afford 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-6,7-dihydro-5H-pteridine (1.7 mg, 2.47%) as an off-white solid. LC-MS (ESI) m / z 523 [M+H] I-19

[0676]

[0677] The I-19 was manufactured in a similar manner to the I-5. 1 H NMR (400MHz, DMSO-d6) δ8.64(s,1H),8.53(s,1H),8.02(d,J=1.4Hz,1H),7.61–7.53(m,2H),7.47(d,J=8.1Hz,2H),5.24(s,2H),5.05(s ,2H),4.05(q,J=7.3Hz,2H),3.81(s,3H),1.68(tt,J=8.2,4.6Hz,1H),1.30(t,J=7.3Hz,3H),0.98(q,J=3.4Hz,2H),0.80–0.72(m,2H). LC-MS(ESI)m / z 552[M+H].

[0678] I-20

[0679] I-20 was synthesized in a similar manner to I-1.

[0680]

[0681] 1 H NMR: (300MHz, DMSO-d6) δ9.31(s,1H),8.70(s,1H),8.48(s,1H),8.22–8.1 2(m,1H),7.45(q,J=8.3Hz,4H),6.91(d,J=9.5Hz,1H),5.58(s,2H),5.05( t,J=7.3Hz,1H),4.03(d,J=8.5Hz,2H),3.94(s,1H),3.84(s,3H),2.63(s, 3H), 1.73 (td, J=7.9, 4.1Hz, 1H), 1.02 (q, J=3.3Hz, 2H), 0.83–0.72 (m, 2H). LC-MS (ESI) m / z 589.05 [M+H].

[0682] I-21

[0683]

[0684] Step 1: 4-[1-cyclopropyl-4-(trifluoromethyl)imidazol-2-yl]benzoate

[0685] Under an oxygen atmosphere at 80 ° C by 4- [4- (trifluoromethyl) -1H- imidazole -2- bases] methyl benzoate (5g, 18.504mmol, 1 equivalent), bipyridine (8670.34mg, 55.512mmol, 3 equivalents), bromocyclopropane (6715.70mg, 55.512mmol, 3 equivalents) and Cu (OAc) 2 (10.08g, 55.512mmol, 3 equivalents) in DCE (50mL) was stirred overnight. The resulting mixture was diluted with DCM (100mL). The resulting mixture was washed with 3x 100mL water. The resulting organic phase was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (10 mmol / L NH4HCO3) over 10 min with UV detection (254 nm) to afford methyl 4-[1-cyclopropyl-4-(trifluoromethyl)imidazol-2-yl]benzoate (430 mg, 7.49%) as a brown solid. LC-MS (ESI) m / z 311.10 [M+H].

[0686] Step 2: {4-[1-cyclopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methanol

[0687] To a stirred solution of 4-[1-cyclopropyl-4-(trifluoromethyl)imidazol-2-yl]benzoic acid methyl ester (160 mg, 0.516 mmol, 1 equivalent) in THF (5 mL) was added diisobutylaluminum hydride (2.58 mL, 2.580 mmol, 5 equivalents, 1 M hexane solution) dropwise under a nitrogen atmosphere at room temperature. The resulting mixture was stirred at room temperature for 1 h under a nitrogen atmosphere. The reaction was quenched with saturated NH4Cl (aqueous solution) at room temperature. The resulting mixture was filtered and the filter cake was washed with water (3 x 10 mL). The aqueous layer was extracted with EtOAc (3 x 10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 10% to 50% MeCN / water (10 mmol / L NH4HCO3) over 10 min with UV detection (254 nm) to afford {4-[1-cyclopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methanol (120 mg, 82.44%) as a brown oil. LC-MS (ESI) m / z 283.10 [M+H].

[0688] Step 3 :2-[4-(chloromethyl)phenyl]-1-cyclopropyl-4-(trifluoromethyl)imidazole

[0689] To a stirred solution of {4-[1-cyclopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methanol (120 mg, 0.425 mmol, 1 eq) in DCE (5 mL) was added thionyl chloride (252.87 mg, 2.125 mmol, 5 eq) dropwise at room temperature. The resulting mixture was stirred at 50 ° C for 20 min under a nitrogen atmosphere. The resulting mixture was diluted with DCM (10 mL). The resulting mixture was washed with water (3 x 10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give 2-[4-(chloromethyl)phenyl]-1-cyclopropyl-4-(trifluoromethyl)imidazole (110 mg, 86.04%) as a brown oil. LC-MS (ESI) m / z 301.06 [M + H].

[0690] Step 4: Synthesis of 8-({4-[1-cyclopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)pyrido[2,3-d]pyrimidin-7-one

[0691] A mixture of 2-[4-(chloromethyl)phenyl]-1-cyclopropyl-4-(trifluoromethyl)imidazole (28 mg, 0.093 mmol, 1 eq), 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8H-pyrido[2,3-d]pyrimidin-7-one (27.50 mg, 0.093 mmol, 1 eq) and 1,1,3,3-tetramethylguanidine (16.09 mg, 0.140 mmol, 1.5 eq) in DMF (2 mL) was stirred at room temperature under nitrogen atmosphere overnight. The crude product was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 10% to 50% MeCN / water (10 mmol / L NH4HCO3) over 10 min with UV detector (254 nm) to give 8-({4-[1-cyclopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)pyrido[2,3-d]pyrimidin-7-one (10 mg, 18.48%) as an off-white solid. 1H NMR: (400MHz, DMSO-d6)δ9.29(d,J=2.6Hz,1H),8.69(s,1H),8.16(dd,J=9.5, 2.6Hz,1H),7.91(d,J=1.4Hz,1H),7.84–7.78(m,2H),7.39(d,J=8.0Hz,2H),6. 91(dd,J=9.5,3.5Hz,1H),5.57(s,2H),3.85–3.79(m,3H),3.70(d,J=4.5Hz,1H ), 1.73 (dd, J = 8.2, 4.0 Hz, 1H), 1.07–0.83 (m, 6H), 0.76 (dt, J = 7.1, 3.5 Hz, 2H). LC-MS (ESI) m / z 560.10 [M+H].

[0692] I-22

[0693]

[0694] Step 1: 2-(4-bromo-2-fluoro-6-methoxyphenyl)-4-(trifluoromethyl)-1H-imidazole

[0695] A mixture of 3,3-dibromo-1,1,1-trifluoropropane-2-one (259.38 mg, 0.961 mmol, 1.12 equivalents) and AcONa (78.85 mg, 0.961 mmol, 1.12 equivalents) in H2O (0.4 mL, 2 equivalents) was stirred at 100 ° C under a nitrogen atmosphere for 1 h. The mixture was cooled to room temperature. 4-bromo-2-fluoro-6-methoxybenzaldehyde (200 mg, 0.858 mmol, 1 equivalent), MeOH (3 mL, 15.2 equivalents) and NH4OH (0.7 mL, 17.976 mmol, 20.95 equivalents) were added to the above mixture at room temperature. The resulting mixture was stirred at room temperature for another 40 min. The resulting mixture was then stirred at 100 ° C under a nitrogen atmosphere for 2 h. The reaction was quenched with water at 0 ° C. The resulting mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (2 x 5 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was dissolved in MeOH (3 mL). The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% MeCN / water (0.1% FA) in 10 min with a UV detector (254 nm) to obtain 2-(4-bromo-2-fluoro-6-methoxyphenyl)-4-(trifluoromethyl)-1H-imidazole (90 mg, 30.93%) as a light yellow solid. LC-MS (ESI) m / z 338.9 [M+H]

[0696] Step 2: 2-(4-Bromo-2-fluoro-6-methoxyphenyl)-1-isopropyl-4-(trifluoromethyl)imidazole

[0697] To a stirred solution of 2-(4-bromo-2-fluoro-6-methoxyphenyl)-4-(trifluoromethyl)-1H-imidazole (90 mg, 0.265 mmol, 1 eq) in dimethylformamide (1 mL, 20 eq) was added cesium sulfate methane peroxyacid (173.49 mg, 0.530 mmol, 2 eq) and 2-iodopropane (135.36 mg, 0.795 mmol, 3 eq) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 65 ° C for 3 h under a nitrogen atmosphere. The resulting mixture was filtered and the filter cake was washed with THF (3 x 5 mL). The filtrate was concentrated under reduced pressure. The residue was dissolved in THF (2 mL). The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give 2-(4-bromo-2-fluoro-6-methoxyphenyl)-1-isopropyl-4-(trifluoromethyl)imidazole (70 mg, 69.19%) as a yellow solid. LC-MS (ESI) m / z 383.1 [M+H]

[0698] Step 3: Methyl 3-fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-5-methoxybenzoate

[0699] To a stirred solution of 2-(4-bromo-2-fluoro-6-methoxyphenyl)-1-isopropyl-4-(trifluoromethyl)imidazole (100 mg, 0.262 mmol, 1 eq) in MeOH (8 mL, 197.591 mmol) was added Pd(dppf)Cl2 (35 mg, 0.048 mmol, 0.18 eq) and TEA (1 mL, 7.194 mmol, 27.42 eq) at room temperature under a carbon monoxide atmosphere. The resulting mixture was stirred at 90 ° C. under a carbon monoxide atmosphere overnight. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in THF (2 mL). The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give methyl 3-fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-5-methoxybenzoate (70 mg, 74.05%) as a yellow solid. LC-MS (ESI) m / z 361.1 [M+H]

[0700] Step 4:{3-Fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-5-methoxyphenyl}methanol

[0701] To a stirred mixture of methyl 3-fluoro-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-5-methoxybenzoate (70 mg, 0.202 mmol, 1 eq) in THF (3 mL, 37.028 mmol, 190.59 eq) was added LiAlH4 (22.12 mg, 0.582 mmol, 3.0 eq) in portions at 0 ° C under a nitrogen atmosphere. The resulting mixture was stirred at 0 ° C for 1 h under a nitrogen atmosphere. The reaction was quenched with 10H2O.Na2SO4 at 0 ° C. The resulting mixture was filtered and the filter cake was washed with THF (2 x 2 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluted with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give {3-fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-5-methoxyphenyl}methanol (50 mg, 77.45%) as a brown solid. LC-MS (ESI) m / z 333.1 [M+H]

[0702] Step 5: 2-[4-(Chloromethyl)-2-fluoro-6-methoxyphenyl]-1-methyl-4-(trifluoromethyl)imidazole

[0703] To a stirred solution of {3-fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-5-methoxyphenyl}methanol (100 mg, 0.301 mmol, 1 equiv) in DCM (2 mL, 0.024 mmol) was added SOCl (107.5 mg, 0.904 mmol, 3.00 equiv) at 50 ° C under a nitrogen atmosphere. The resulting mixture was stirred at 50 ° C for 30 min under a nitrogen atmosphere. The reaction was quenched with water / ice at 0 ° C. The resulting mixture was extracted with CHCl (3 x 5 mL). The combined organic layers were washed with brine (3 x 5 mL), dried over anhydrous NaSO, filtered and concentrated under reduced pressure to give 2-[4-(chloromethyl)-2-fluoro-6-methoxyphenyl]-1-methyl-4-(trifluoromethyl)imidazole (110 mg, 113.28%) as a brown solid. The crude product was used directly in the next step without further purification. LC-MS (ESI) m / z 351.1 [M+H]

[0704] Step 6:2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({3-fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-5-methoxyphenyl}methyl)pyrido[2,3-d]pyrimidin-7-one

[0705] A solution of 2-[4-(chloromethyl)-2-fluoro-6-methoxyphenyl]-1-isopropyl-4-(trifluoromethyl)imidazole (21.3 mg, 0.061 mmol, 1 equiv), 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8H-pyrido[2,3-d]pyrimidin-7-one (17.93 mg, 0.061 mmol, 1 equiv) and 1,1,3,3-tetramethylguanidine (10.49 mg, 0.091 mmol, 1.5 equiv) in DMF (2 mL, 25.843 mmol, 425.55 equiv) was stirred at room temperature under nitrogen atmosphere overnight. The crude product was purified by preparative HPLC under the following conditions (column: Xselect CSH F-phenyl OBD column, 19*250 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: MeOH--HPLC; flow rate: 25 mL / min; gradient: 60% B to 80% B, 80% B in 8 min; wavelength: 220 nm; RT1 (min): 7.56;) to give 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({3-fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-5-methoxyphenyl}methyl)pyrido[2,3-d]pyrimidin-7-one (9 mg, 23.41%) as an off-white solid. 1 H NMR (400MHz, DMSO-d6) δ9.30 (s, 1H), 8.69 (s, 1H), 8.18 (t, J = 4.8Hz, 2H), 7.04 ( s,1H),6.92(d,J=9.5Hz,1H),6.72(d,J=9.6Hz,1H),5.56(s,2H),3.91(p,J=6.9 Hz,1H),3.82(s,3H),3.69(s,3H),1.75(dq,J=8.7,5.2,4.4Hz,1H),1.35(d,J= 6.7Hz, 3H), 1.23 (d, J = 6.7Hz, 3H), 1.02 (d, J = 4.6Hz, 2H), 0.77 (d, J = 7.5Hz, 2H). LC-MS (ESI) m / z 610.05 [M+H].

[0706] I-23

[0707]

[0708] Step 1:2-(4-bromo-2-fluoro-3-methoxyphenyl)-4-(trifluoromethyl)-1H-imidazole

[0709] A mixture of 3,3-dibromo-1,1,1-trifluoropropane-2-one (1.30 g, 4.806 mmol, 1.12 equiv) and NaOAc (0.97 g, 11.879 mmol, 1.12 equiv) in H2O (2.00 mL, 111.008 mmol, 25.87 equiv) was stirred at 100 ° C under a nitrogen atmosphere for 1 h. 4-bromo-2-fluoro-3-methoxybenzaldehyde (1 g, 4.291 mmol, 1 equiv) and MeOH (30 mL, 15.2 equiv) and NH3-H2O (3.50 mL, 89.896 mmol, 20.95 equiv) were added to the above mixture at room temperature. The resulting mixture was stirred at room temperature for another 40 min. The resulting mixture was stirred at 100 ° C under a nitrogen atmosphere for 2 h. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc (3x 10mL). The combined organic layer was washed with water (3x 5mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% MeCN / water (10mmol / L NH4HCO3) in 10min with a UV detector (254nm) to obtain 2-(4-bromo-2-fluoro-3-methoxyphenyl)-4-(trifluoromethyl)-1H-imidazole (880mg, 60.48%) as a yellow solid. LC-MS (ESI) m / z 339[M+H]

[0710] Step 2: 2-(4-bromo-2-fluoro-3-methoxycyclohexyl)-1-isopropyl-4-(trifluoromethyl)imidazolidine

[0711] To a stirred mixture of 2-(4-bromo-2-fluoro-3-methoxycyclohexyl)-4-(trifluoromethyl)imidazolidine (880 mg, 2.520 mmol, 1 eq) and CsCO (1642.31 mg, 5.040 mmol, 2 eq) in DMF (4 ml) was added 2-iodopropane (1285.29 mg, 7.560 mmol, 3 eq) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 65 ° C for 2 h. The resulting mixture was filtered and the filter cake was washed with MeCN (1 x 1 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluted with 10% to 50% MeCN / water (10 mmol / L NH4HCO3) over 10 min with UV detection (254 nm) to afford 2-(4-bromo-2-fluoro-3-methoxycyclohexyl)-1-isopropyl-4-(trifluoromethyl)imidazolidine (880 mg, 89.24%) as a yellow solid. LC-MS (ESI) m / z 381 [M+H]

[0712] Step 3: Methyl 3-fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-2-methoxybenzoate

[0713] To a stirred mixture of 2-(4-chloro-2-fluoro-3-methoxyphenyl)-1-isopropyl-4-(trifluoromethyl)imidazole (860 mg, 2.554 mmol, 1 eq) and TEA (1550.76 mg, 15.324 mmol, 6 eq) in MeOH (16.00 mL, 395.181 mmol, 154.72 eq) was added Pd(dppf)Cl2 (186.89 mg, 0.255 mmol, 0.1 eq) at room temperature under a carbon monoxide atmosphere. The resulting mixture was stirred at 90 ° C for an additional 12 h. The resulting mixture was filtered and the filter cake was washed with MeOH (3 x 4 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluted with 10% to 50% MeCN / water (10 mmol / L NH4HCO3) over 10 min with UV detection (254 nm) to give methyl 3-fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-2-methoxybenzoate (600 mg, 65.20%) as a yellow oil. LC-MS (ESI) m / z 361 [M+H]

[0714] Step 4: {3-Fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-2-methoxyphenyl}methanol

[0715] To a stirred mixture of methyl 3-fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-2-methoxybenzoate (600 mg, 1.665 mmol, 1 eq) in THF (8.82 mL, 108.891 mmol, 65.40 eq) was added LiAlH4 (189.59 mg, 4.995 mmol, 3 eq) in portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2 h. The reaction was quenched with 10H2O.Na2SO4 at 0°C. The resulting mixture was filtered and the filter cake was washed with THF (2 x 2 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE / EA (1:1)) to give {3-fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-2-methoxyphenyl}methanol (320 mg, 57.83%) as a white oil. LC-MS (ESI) m / z 333 [M+H]

[0716] Step 5: 2-[4-(Chloromethyl)-2-fluoro-3-methoxyphenyl]-1-isopropyl-4-(trifluoromethyl)imidazole

[0717] To a stirred mixture of {3-fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-2-methoxyphenyl}methanol (30 mg, 0.090 mmol, 1 eq) in DCM (1 mL, 15.731 mmol, 174.24 eq) was added SOCl2 (0.02 mL, 0.270 mmol, 3 eq) at room temperature under a nitrogen atmosphere. The reaction was quenched with saturated NaHCO3 (aqueous solution) and extracted with CH2Cl2 (3 x mL). The combined organic layers were washed with water (2 x 2 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The crude product was used directly in the next step without further purification. LC-MS (ESI) m / z 351 [M+H]

[0718] Step 6 :2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({3-fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-2-methoxyphenyl}methyl)pyrido[2,3-d]pyrimidin-7-one

[0719] To a stirred mixture of 2-[4-(chloromethyl)-2-fluoro-3-methoxyphenyl]-1-isopropyl-4-(trifluoromethyl)imidazole (40 mg, 0.114 mmol, 0.9 eq) and 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8H-pyrido[2,3-d]pyrimidin-7-one (37.42 mg, 0.127 mmol, 1 eq) in DMF (4.44 mL, 57.407 mmol, 453.21 eq) was added 1,1,3,3-tetramethylguanidine (21.89 mg, 0.190 mmol, 1.5 eq) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN in water (0.1% FA) over 10 min with UV detector (254 nm) to give 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({3-fluoro-4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]-2-methoxyphenyl}methyl)pyrido[2,3-d]pyrimidin-7-one (20 mg, 25.89%) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ9.31(s,1H),8.66(s,1H),8.25–8.16(m,2H),7.08(t,J=7.3Hz,1H),6.93(d,J=9.6Hz,1H),6.70(d,J=8.1Hz,1H),5.59 (s,2H),4.17–4.09(m,1H),3.89(s,3H),3.78(s,3H),1.74–1.67(m,1H) ,1.35(d,J=6.6Hz,6H),1.01–0.95(m,2H),0.70(dd,J=7.9,3.4Hz,2H). LC-MS(ESI)m / z 610.25[M+H]

[0720] I-24

[0721]

[0722] Step 1: 2-Chloro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyrimidine-5-carbonitrile

[0723] To a stirred solution of 2,4-dichloropyrimidine-5-carbonitrile (1.7 g, 9.771 mmol, 1 eq) and 1-{4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methanamine (2.77 g, 9.771 mmol, 1 eq) in THF (20 mL) was added TEA (1.98 g, 19.542 mmol, 2 eq) dropwise at room temperature. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 2 h. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give 2-chloro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyrimidine-5-carbonitrile (900 mg, 21.89%) as an off-white solid. LC-MS (ESI) m / z 421.8 [M+H]

[0724] Step 2: 4'-Cyclopropyl-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]-6'-methoxy-[2,5'-bipyrimidine]-5-carbonitrile

[0725] To a stirred solution of 2-chloro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyrimidine-5-carbonitrile (438 mg, 1.041 mmol, 1 eq) and 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (302.88 mg, 1.561 mmol, 1.5 eq) in H2O (0.5 mL) and dioxane (5 mL) was added K3PO4 (552.33 mg, 2.603 mmol, 2.5 eq) and Pd(dppf)Cl2 (152.32 mg, 0.208 mmol, 0.2 eq) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 90° C. under a nitrogen atmosphere for 2 hours. The resulting mixture was filtered, and the filter cake was washed with THF (3×5 mL) and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give 4'-cyclopropyl-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]-6'-methoxy-[2,5'-bipyrimidine]-5-carbonitrile (422.4 mg, 75.92%) as a light yellow solid. LC-MS (ESI) m / z 535.5 [M+H]

[0726] Step 35-(Aminomethyl)-4'-cyclopropyl-N-({4-[4-(difluoromethyl)-1-isopropylimidazol-2-yl]phenyl}methyl)-6'-methoxy-[2,5'-bipyrimidinyl]-4-amine

[0727] To a stirred solution of 4'-cyclopropyl-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]-6'-methoxy-[2,5'-bipyrimidine]-5-carbonitrile (422.4 mg, 0.790 mmol, 1 eq) in THF (5 mL) was added LiAlH4 (89.96 mg, 2.370 mmol, 3 eq) at 0°C under a nitrogen atmosphere. The resulting mixture was stirred at 0°C for 1 hour under a nitrogen atmosphere. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EA (3 x 10 mL), dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 10% to 50% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give 5-(aminomethyl)-4'-cyclopropyl-N-({4-[4-(difluoromethyl)-1-isopropylimidazol-2-yl]phenyl}methyl)-6'-methoxy-[2,5'-bipyrimidinyl]-4-amine (161.5 mg, 39.26%) as a colorless oil. LC-MS (ESI) m / z 539.5 [M+H]

[0728] Step 4: 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-3H,4H-pyrimido[4,5-d][1,3]diazin-2-one

[0729] To a stirred solution of 5-(aminomethyl)-4'-cyclopropyl-N-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-6'-methoxy-[2,5'-bipyrimidinyl]-4-amine (187 mg, 0.347 mmol, 1 eq) in DMF (2 mL) was added NaH (16.66 mg, 0.694 mmol, 2 eq) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 5 min under a nitrogen atmosphere. To the above mixture was added CDI (112.60 mg, 0.694 mmol, 2 eq) at room temperature. The resulting mixture was stirred at room temperature overnight. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EA (3 x 10 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN in water (0.1% FA) over 10 min with UV detector (254 nm) to give 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-3H,4H-pyrimido[4,5-d][1,3]diazin-2-one (11.7 mg, 5.89%) as an off-white solid. 1 H NMR: (400MHz, DMSO-d6) δ8.70(s,1H),8.62(s,1H),8.23(s,1H),7.78(s,1H),7.54(d,J=7.9Hz,2H),7.46(d,J=7.9Hz,2H),5.24(s,2H ),4.60(s,2H),4.56–4.45(m,1H),3.87(s,3H),1.71(s,1H),1.46(d,J=6.6Hz,6H),1.02(d,J=4.5Hz,2H),0.79(dd,J=8.0,3.8Hz,2H). LC-MS(ESI)m / z565.10[M+H]

[0730] I-25

[0731]

[0732] To a stirred solution of 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-3H,4H-pyrimido[4,5-d][1,3]diazin-2-one (16.7 mg, 0.030 mmol, 1 eq) in DMF (2 mL) was added NaH (1.42 mg, 0.060 mmol, 2 eq) at 0°C under a nitrogen atmosphere. The resulting mixture was stirred at 0°C for 30 min under a nitrogen atmosphere. To the above mixture was added MeI (12.60 mg, 0.090 mmol, 3 eq) dropwise at 0°C. The resulting mixture was stirred at room temperature for another 2 h. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EA (2 x 10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column, eluting with 10% to 50% MeCN in water (0.1% FA) over 10 min with UV detector (254 nm) to give 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-3-methyl-4H-pyrimido[4,5-d][1,3]diazin-2-one (7.1 mg, 37.71%) as an off-white solid. 1 H NMR (400MHz, DMSO-d6) δ8.60(d,J=25.0Hz,2H),8.15(s,1H),7.52–7.34(m,4H),5.20(s,2H),4.62(s,2H),4.43(p,J=6 .8Hz,1H),3.80(s,3H),2.99(s,3H),1.64(p,J=7.5,5.9Hz,1H),1.39(d,J=6.6Hz,6H),0.96(s,2H),0.78–0.67(m,2H). LC-MS(ESI)m / z579.05[M+H]

[0733] I-25

[0734]

[0735] Step 1: 4-[4-(Trifluoromethyl)-1H-imidazol-2-yl]benzonitrile

[0736] To a stirred solution of 3,3-dibromo-1,1,1-trifluoropropane-2-one (23.05 g, 85.409 mmol, 1.12 equivalents) in H2O (20 mL) was added AcONa (7.01 g, 85.409 mmol, 1.12 equivalents) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 100 ° C for 1 h under a nitrogen atmosphere and then cooled to room temperature. A solution of 4-formyl-benzonitrile (10 g, 76.258 mmol, 1 equivalent) in MeOH (152 mL) and NH3H2O ​​(35.6 mL) was added to the above mixture. The resulting mixture was stirred at room temperature for 40 min. The reaction was then warmed to 100 ° C and stirred for 2 h. The reaction was quenched by adding water / ice (50 mL) at room temperature. The resulting mixture was extracted with EA (2 x 60 mL), dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE / EA (5:1)) to give 4-[4-(trifluoromethyl)-1H-imidazol-2-yl]benzonitrile (13.8 g, 76.30%) as a yellow solid. LC-MS (ESI) m / z 238.1 [M+H]

[0737] Step 2: 4-[1-Isopropyl-4-(trifluoromethyl)imidazol-2-yl]benzonitrile

[0738] To a stirred solution of 4-[4-(trifluoromethyl)-1H-imidazol-2-yl]benzonitrile (5 g, 21.081 mmol, 1 eq) in DMF (80 mL) was added CsCO (13.74 g, 42.162 mmol, 2 eq) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2 min under a nitrogen atmosphere. 2-Iodopropane (10.75 g, 63.243 mmol, 3 eq) was added dropwise to the above mixture at room temperature. The resulting mixture was stirred at 60 ° C. overnight. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EA (3 x 60 mL). The combined organic layers were washed with brine (3 x 60 mL), dried over anhydrous NaSO and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE / EA (5:1)) to give 4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]benzonitrile (3.3 g, 56.05%) as a yellow solid. LC-MS (ESI) m / z 280.2 [M+H]

[0739] Step 3: 1-{4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methanamine

[0740] To a stirred solution of 4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]benzonitrile (3.5 g, 12.533 mmol, 1 equivalent) in EA (40 mL) and NH H O (20 mL) was added Raney nickel (3.96 g, 46.264 mmol, 3.4 equivalents) at room temperature under an air atmosphere. The resulting mixture was stirred for 5 h at room temperature under a hydrogen atmosphere. The resulting mixture was filtered, the filter cake was washed with EA (3x20 mL) and concentrated under reduced pressure. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 0% to 100% MeCN / water (0.1% FA) in 10 min with a UV detector (254 nm) to obtain 1-{4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methylamine (3.3 g, 85.61%) as an off-white solid. 1 H NMR (400MHz, DMSO-d6) δ 8.44 (s, 1H), 8.19 (d, J = 1.4Hz, 1H), 7.59 (s, 5H), 4.47 (p, J = 6.6Hz, 1H), 4.02 (s, 2H), 1.41 (d, J = 6.7Hz, 6H). LC-MS(ESI)m / z 284.2[M+H]

[0741] Step 4: 2-Chloro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyrimidine-5-carboxamide

[0742] To a stirred solution of 2,4-dichloropyrimidine-5-carboxamide (638 mg, 3.323 mmol, 1 eq) and 1-{4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methanamine (941.38 mg, 3.323 mmol, 1 eq) in THF (5 mL) was added TEA (672.51 mg, 6.646 mmol, 2 eq) dropwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2 h under a nitrogen atmosphere. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detection (254 nm) to give 2-chloro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyrimidine-5-carboxamide (780 mg, 53.49%) as an off-white solid. LC-MS (ESI) m / z 439.8 [M+H]

[0743] Step 5:4'-Cyclopropyl-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]-6'-methoxy-[2,5'-bipyrimidine]-5-carboxamide

[0744] To a stirred solution of 2-chloro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyrimidine-5-carboxamide (400 mg, 0.911 mmol, 1 eq) and 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (265.24 mg, 1.367 mmol, 1.5 eq) in dioxane (4 mL) and H2O (0.5 mL) was added K3PO4 (483.70 mg, 2.277 mmol, 2.5 eq) and Pd(dppf)Cl2 (133.39 mg, 0.182 mmol, 0.2 eq) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 90°C under nitrogen atmosphere for 2 hours. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 10% to 50% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give 4'-cyclopropyl-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]-6'-methoxy-[2,5'-bipyrimidine]-5-carboxamide (485 mg, 96.30%) as a light yellow solid. LC-MS (ESI) m / z 553.5 [M+H]

[0745] Step 6: 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-3H-pyrimido[4,5-d][1,3]diazine-2,4-dione

[0746] To a stirred solution of 4'-cyclopropyl-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]-6'-methoxy-[2,5'-bipyrimidine]-5-carboxamide (435 mg, 0.787 mmol, 1 eq) in DMF (5 mL) was added NaH (37.78 mg, 1.574 mmol, 2 eq) at 0°C under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 5 min under a nitrogen atmosphere. To the above mixture was added CDI (255.31 mg, 1.574 mmol, 2 eq) at room temperature. The resulting mixture was stirred at room temperature for an additional 2 h. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EA (3 x 15 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 10% to 50% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to afford 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-3H-pyrimido[4,5-d][1,3]diazine-2,4-dione (370 mg, 81.24%) as an off-white solid. LC-MS (ESI) m / z 579.5 [M+H]

[0747] Step 7: 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-3-methylpyrimido[4,5-d][1,3]diazine-2,4-dione

[0748] To a stirred solution of 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-3H-pyrimido[4,5-d][1,3]diazine-2,4-dione (30 mg, 0.052 mmol, 1 eq) in DMF (1 mL) was added KCO (14.33 mg, 0.104 mmol, 2 eq) and MeI (14.72 mg, 0.104 mmol, 2 eq) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at room temperature under nitrogen atmosphere overnight. The residue was purified by reverse phase flash chromatography (C18 silica gel column, eluting with 10% to 50% MeCN in water (0.1% FA) over 10 min with UV detector (254 nm) to give 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-3-methylpyrimido[4,5-d][1,3]diazine-2,4-dione (19.7 mg, 63.73%) as an off-white solid. 1 H NMR: (400MHz, DMSO-d6) δ9.37(s,1H),8.69(s,1H),8.17(s,1H),7.50(t,J=6.4Hz,4H),5.44(s,2H),4.42(p,J=6.7Hz,1H),3. 83(s,3H),3.34(s,3H),1.72(tt,J=8.5,4.7Hz,1H),1.39(d,J=6.6Hz,6H),1.01(p,J=3.5Hz,2H),0.75(dq,J=7.3,3.6Hz,2H). LC-MS (ESI) m / z 593.10 [M+H].

[0749] I-27

[0750]

[0751] Step 1: Ethyl 6-chloro-5-fluoro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridine-3-carboxylate

[0752] To a stirred solution of ethyl 4,6-dichloro-5-fluoropicolinate (900 mg, 3.781 mmol, 1 eq) and TEA (1147.79 mg, 11.343 mmol, 3 eq) in THF (10 mL) was added 1-{4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methanamine (1071.11 mg, 3.781 mmol, 1 eq) dropwise at 0° C. The resulting mixture was stirred at room temperature overnight and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give ethyl 6-chloro-5-fluoro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridine-3-carboxylate (450 mg, 24.55%) as a yellow solid. LC-MS (ESI) m / z 485.8 [M+H].

[0753] Step 2: {6-chloro-5-fluoro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridin-3-yl}methanol

[0754] To a stirred solution of ethyl 6-chloro-5-fluoro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridine-3-carboxylate (400 mg, 0.825 mmol, 1 eq) in THF (5 mL) was added LiAlH (40.70 mg, 1.073 mmol, 1.3 eq) in portions at 0°C. The resulting mixture was stirred at 0°C under a nitrogen atmosphere for 2 h. The resulting mixture was filtered and the filter cake was washed with THF (3 x 10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 10% to 50% MeCN / water (0.1% FA) over 10 min with UV detection (254 nm) to give {6-chloro-5-fluoro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridin-3-yl}methanol (200 mg, 54.7%) as a brown solid. LC-MS (ESI) m / z 443.8 [M+H].

[0755] Step 3: 6-Chloro-5-fluoro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridine-3-carbaldehyde

[0756] To a stirred solution of {6-chloro-5-fluoro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridin-3-yl}methanol (200 mg, 0.452 mmol, 1 eq) in DCM (2 mL) was added DMP (383.11 mg, 0.904 mmol, 2 eq) at room temperature. The resulting mixture was stirred at room temperature for 2 h and concentrated under reduced pressure. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 10% to 100% MeCN / water (0.1% FA) over 10 min with a UV detector (254 nm) to afford 6-chloro-5-fluoro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridine-3-carbaldehyde (150 mg, 75.34%) as a light yellow solid. LC-MS (ESI) m / z: 441.1 [M+H]

[0757] Step 4: 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-5-fluoro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridine-3-carbaldehyde

[0758] To a stirred solution of 6-chloro-5-fluoro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridine-3-carbaldehyde (50 mg, 0.113 mmol, 1 eq) and 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (33.01 mg, 0.170 mmol, 1.5 eq) in 1,4-dioxane (1 mL) and H2O (0.2 mL) was added Pd(dppf)Cl2 (16.60 mg, 0.023 mmol, 0.2 eq) and K3PO4 (60.19 mg, 0.283 mmol, 2.5 eq) at room temperature. The resulting mixture was stirred at 90 ° C under a nitrogen atmosphere for 2 hours. The resulting mixture was filtered, and the filter cake was washed with 1,4-dioxane (3 x 10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 10% to 50% MeCN / water (0.1% TFA) over 10 min with UV detection (254 nm) to afford 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-5-fluoro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyridine-3-carbaldehyde (35 mg, 55.6%) as an off-white solid. LC-MS (ESI) m / z 555.5 [M+H].

[0759] Step 5:7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-fluoro-1-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-1,6-naphthyridin-2-one

[0760] To a stirred solution of ethyl acetate (79.44 mg, 0.900 mmol, 10 equivalents) in THF (2 mL) was added LiHMDS (0.90 mL, 0.900 mmol, 10 equivalents, 1 M THF solution) dropwise at -78 ° C under a nitrogen atmosphere. The resulting mixture was stirred at -78 ° C for 10 min under a nitrogen atmosphere. Next, a solution of 6- (4- cyclopropyl -6- methoxypyrimidin-5-yl) -5-fluoro-4- [( {4- [1- isopropyl -4- (trifluoromethyl) imidazole -2- yl] phenyl} methyl) amino] pyridine -3- carbaldehyde (50 mg, 0.090 mmol, 1 equivalent) in THF (1 mL) was added to the reaction mixture at -78 ° C. The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere and concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column, eluting with 10% to 100% MeCN in water (0.1% FA) over 15 min with UV detector (254 nm) to give 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-fluoro-1-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-1,6-naphthyridin-2-one (16.6 mg, 31.6%) as an off-white solid. 1 H NMR (400MHz, DMSO-d6) δ8.92(s,1H),8.66(s,1H),8.26–8.19(m,1H),8.15(s,1H),7.51(d,J=8.0Hz,2H),7.27(d,J=8.0Hz,2H),6.96(d,J=9.5Hz, 1H),5.69–5.64(m,2H),4.43(p,J=6.6Hz,1H),3.73(s,3H),1.60(s,1H), 1.38(d,J=6.6Hz,6H),1.11–1.03(m,1H),0.92–0.81(m,2H),0.70(s,1H). LC-MS(ESI)m / z 579.1[M+H]

[0761] I-28

[0762]

[0763] The I-28 was manufactured in a similar manner to the I-17. 1H NMR (400MHz, DMSO-d6) δ9.38(d,J=1.1Hz,1H),8.70(s,1H),8.48(s,1H),8.02(s,1H),7.56(d,J=7.9Hz,2H),7.49(d,J=8.0H z,2H),5.49(s,2H),4.04(q,J=7.2Hz,2H),3.91–3.80(m,3H),1.76(s,1H),1.29(t,J=7.2Hz,3H),1.03(s,2H),0.78(s,2H). LC-MS(ESI)m / z 549.0[M+H]

[0764] I-29

[0765]

[0766] The I-29 was manufactured in a similar manner to the I-17. 1 H NMR(300MHz,DMSO-d6)δ9.38(s,1H),8.71(s,1H),8.49(s,1H),8.18(d,J=1.4Hz,1H),7.50(s,4H),5.51(s,2H) ,4.41(p,J=6.7Hz,1H),3.85(s,3H),1.75(m,1H),1.39(d,J=6.6Hz,6H),1.10–0.99(m,2H),0.82–0.70(m,2H). LC-MS(ESI)m / z 563.1[M+H].

[0767] I-30

[0768]

[0769] Step 1: 6-Bromo-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8H-pyrido[2,3-d]pyrimidin-7-one

[0770] To a stirred solution of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8H-pyrido[2,3-d]pyrimidin-7-one (1 g, 3.386 mmol, 1 eq) and NBS (1.81 g, 10.158 mmol, 3 eq) in DMF (15 mL) was added BPO (0.17 g, 0.677 mmol, 0.2 eq) at room temperature. The resulting mixture was stirred at room temperature under a nitrogen atmosphere overnight and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% TFA) over 10 min with UV detector (254 nm) to afford 6-bromo-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8H-pyrido[2,3-d]pyrimidin-7-one (900 mg, 71.0%) as a yellow solid. LC-MS (ESI) m / z 374.2 [M+H].

[0771] Step 2: 6-Bromo-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-ethyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one

[0772] To a stirred solution of 6-bromo-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8H-pyrido[2,3-d]pyrimidin-7-one (150 mg, 0.401 mmol, 1 eq) and 2-[4-(chloromethyl)phenyl]-1-ethyl-4-(trifluoromethyl)imidazole (104.15 mg, 0.361 mmol, 0.9 eq) in DMF (2 mL) was added 1,1,3,3-tetramethylguanidine (69.25 mg, 0.602 mmol, 1.5 eq) dropwise at room temperature. The resulting mixture was stirred at room temperature under a nitrogen atmosphere overnight. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to afford 6-bromo-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-ethyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one (150 mg, 59.7%) as a yellow solid. LC-MS (ESI) m / z 626.4 [M+H].

[0773] Step 3: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-ethyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-6-(4-methylpiperazine-1-carbonyl)pyrido[2,3-d]pyrimidin-7-one

[0774] To a stirred solution of 6-bromo-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-ethyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one (50 mg, 0.080 mmol, 1 eq) and 1-methyl-piperazine (31.98 mg, 0.320 mmol, 4 eq) in THF (4 mL) was added TEA (40.38 mg, 0.400 mmol, 5 eq) and Pd(dppf)Cl2 (11.68 mg, 0.016 mmol, 0.2 eq) at room temperature. The resulting mixture was stirred at 65 ° C under a carbon monoxide atmosphere for 6 h. The resulting mixture was filtered and the filter cake was washed with 1,4-dioxane (3 x 20 mL). The filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC under the following conditions (column: YMC-Actus Triart C18, 30*150 mm, 5 μm; mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 25% B to 45% B, 45% B in 7 min; wavelength: 254 / 220 nm; RT1 (min): 6.18) to give 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-ethyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-6-(4-methylpiperazine-1-carbonyl)pyrido[2,3-d]pyrimidin-7-one (22.3 mg, 40.1%) as an off-white solid. 1 H NMR (400MHz, DMSO-d6) δ9.36 (s, 1H), 8.71 (s, 1H), 8.30 (s, 1H), 8.02 (d, J = 1.4Hz, 1H),7.55(d,J=8.2Hz,2H),7.44(d,J=8.1Hz,2H),5.60(s,2H),4.56(s,1H),4.05( q,J=7.2Hz,2H),3.84(s,4H),3.54(s,1H),3.37(s,2H),3.10(d,J=42.9Hz,3H),2. 86(s,3H),1.72(m,1H),1.29(t,J=7.3Hz,3H),1.03(t,J=4.0Hz,2H),0.77(m,2H). LC-MS (ESI) m / z: 674.3 [M+H] I-31

[0775]

[0776] Step 1: 4-[1-(2H3)methyl-4-(trifluoromethyl)imidazol-2-yl]benzoate

[0777] To a stirred mixture of methyl 4-[4-(trifluoromethyl)-1H-imidazol-2-yl]benzoate (2 g, 7.402 mmol, 1 eq) and CsCO (4.82 g, 14.804 mmol, 2 eq) in DMF (40 mL, 516.862 mmol, 69.83 eq) was added CD3I (1.07 g, 7.402 mmol, 1 eq) dropwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 60 ° C. under a nitrogen atmosphere overnight. The resulting mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (3 x 10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give methyl 4-[1-(2H3)methyl-4-(trifluoromethyl)imidazol-2-yl]benzoate (1.6 g, 75.25%) as a yellow solid. LC-MS (ESI) m / z 288.1 [M+H]

[0778] Step 2: {4-[1-(2H3)methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methanol

[0779] To a stirred mixture of methyl 4-[1-(2H3)methyl-4-(trifluoromethyl)imidazol-2-yl]benzoate (2.3 g, 8.007 mmol, 1 eq) in THF (45 mL, 555.425 mmol, 69.37 eq) was added dropwise AlH(Bu-i)2 (24.02 mL, 24.021 mmol, 3 eq) at 0°C under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2 h under a nitrogen atmosphere. The reaction was quenched with saturated NH4Cl (aqueous solution) at 0°C. The resulting mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (3 x 10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to afford {4-[1-(2H3)methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methanol (1.6 g, 77.08%) as an off-white solid. LC-MS (ESI) m / z 260.1 [M+H]

[0780] Step 3: 2-[4-(Chloromethyl)phenyl]-1-(2H3)methyl-4-(trifluoromethyl)imidazole

[0781] To a stirred mixture of {4-[1-(2H3)methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methanol (1.5 g, 5.786 mmol, 1 eq) in DCE (30 mL, 378.979 mmol, 65.50 eq) was added thionyl chloride (2.06 g, 17.358 mmol, 3 eq) dropwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 50 ° C. under a nitrogen atmosphere for 30 min. The resulting mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (3 x 10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give 2-[4-(chloromethyl)phenyl]-1-(2H3)methyl-4-(trifluoromethyl)imidazole (1.6 g, 99.58%) as an off-white solid. LC-MS (ESI) m / z 254.06 [M+H]

[0782] Step 4: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-(2H3)methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one

[0783] A mixture of 2-[4-(chloromethyl)phenyl]-1-(2H3)methyl-4-(trifluoromethyl)imidazole (36.39 mg, 0.131 mmol, 0.9 equiv), 1,1,3,3-tetramethylguanidine (25.16 mg, 0.219 mmol, 1.5 equiv) and 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8H-pyrido[2,3-d]pyrimidin-7-one (43 mg, 0.146 mmol, 1.00 equiv) in DMF (1 mL, 12.922 mmol, 88.74 equiv) was stirred at room temperature under nitrogen atmosphere overnight. The residue was purified by reverse phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN in water (0.1% FA) over 10 min with UV detector (254 nm) to give 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-({4-[1-(2H3)methyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)pyrido[2,3-d]pyrimidin-7-one (36.3 mg, 45.91%) as an off-white solid. 1H NMR (400MHz, DMSO-d6) δ9.29(s,1H),8.69(s,1H),8.16(d,J=9.6Hz,1H),7.91(d,J=1.4Hz,1H),7.66–7.58(m,2H),7.40(d,J=8.1Hz,2H ), 6.90 (d, J = 9.5Hz, 1H), 5.57 (s, 2H), 3.83 (s, 3H), 1.73 (dq, J = 8.1, 4.1, 3.6Hz, 1H), 1.01 (p, J = 3.8Hz, 2H), 0.76 (dq, J = 6.9, 3.5Hz, 2H). LC-MS(ESI)m / z 537.2[M+H]

[0784] I-32

[0785]

[0786] Step 1 2-Chloro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyrimidine-5-carboxylate

[0787] To a stirred mixture of methyl 2,4-dichloropyrimidine-5-carboxylate (900 mg, 4.348 mmol, 1 eq) and 1-{4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methanamine (1231.67 mg, 4.348 mmol, 1.0 eq) in THF (10 mL) was added TEA (1319.84 mg, 13.044 mmol, 3.0 eq) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 2 h. The resulting mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (2 x 10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give methyl 2-chloro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyrimidine-5-carboxylate (820 mg, 41.56%) as a white solid. LC-MS (ESI) m / z 454.1 [M+H]

[0788] Step 2 :4'-cyclopropyl-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]-6'-methoxy-[2,5'-bipyrimidine]-5-carboxylic acid methyl ester

[0789] To a stirred mixture of methyl 2-chloro-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]pyrimidine-5-carboxylate (130 mg, 0.286 mmol, 1 eq) and 4-cyclopropyl-6-methoxypyrimidin-5-ylboronic acid (83.35 mg, 0.429 mmol, 1.5 eq) in 1,4-dioxane (2.7 mL, 20 eq) and H2O (0.3 mL, 20 eq) was added pd(dppf)Cl2 (20.96 mg, 0.029 mmol, 0.1 eq) and K3PO4 (182.40 mg, 0.858 mmol, 3.0 eq) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 90° C. for 2 hours under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give methyl 4'-cyclopropyl-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]-6'-methoxy-[2,5'-bipyrimidine]-5-carboxylate (150 mg, 92.27%) as a yellow solid. LC-MS (ESI) m / z 568.3 [M+H]

[0790] Step 3: {4'-cyclopropyl-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]-6'-methoxy-[2,5'-bipyrimidinyl]-5-yl}methanol

[0791] To a stirred solution of methyl 4'-cyclopropyl-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]-6'-methoxy-[2,5'-bipyrimidine]-5-carboxylate (75 mg, 0.132 mmol, 1 eq) in tetrahydrofuran (1.5 mL, 20 eq) was added LiAlH4 (15.04 mg, 0.396 mmol, 3.0 eq) in portions at 0°C under a nitrogen atmosphere. The resulting mixture was stirred at 0°C for 1 h under a nitrogen atmosphere. The precipitated solid was collected by filtration and washed with THF (3 x 5 mL). The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 0% to 100% MeCN / water (0.1% FA) over 10 min with UV detector (254 nm) to give {4'-cyclopropyl-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]-6'-methoxy-[2,5'-bipyrimidinyl]-5-yl}methanol (50 mg, 70.13%) as a yellow solid. LC-MS (ESI) m / z 540.3 [M+H]

[0792] Step 4: 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-4H-pyrimido[4,5-d][1,3]oxazin-2-one

[0793] To a stirred mixture of {4'-cyclopropyl-4-[({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)amino]-6'-methoxy-[2,5'-bipyrimidinyl]-5-yl}methanol (50 mg, 0.093 mmol, 1 eq) and NaH (6.67 mg, 0.279 mmol, 3.0 eq) in DMF (1 mL, 20 eq) was added CDI (18.03 mg, 0.112 mmol, 1.2 eq) in portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature overnight under a nitrogen atmosphere. The reaction was quenched with water at 0°C. The resulting mixture was concentrated under reduced pressure. The crude product (50 mg) was purified by preparative HPLC under the following conditions (column: YMC-Actus Triart C18, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 47% B to 67% B, 67% B in 7 min; wavelength: 254 / 220 nm; RT1 (min): 6.53; number of runs: 0) to give 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-({4-[1-isopropyl-4-(trifluoromethyl)imidazol-2-yl]phenyl}methyl)-4H-pyrimido[4,5-d][1,3]oxazin-2-one (12 mg, 22.85%) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ8.71(d,J=1.1Hz,1H),8.65(s,1H),8.17(d,J=1.5Hz,1H),7.47(q,J=8.3Hz,4H),5.58(s,2H),5.22(s,2H),4.42( q, J=6.5Hz, 1H), 3.81 (s, 3H), 1.67 (dq, J=8.5, 4.8, 4.4Hz, 1H), 1.39 (d, J=6.6Hz, 6H), 0.98 (d, J=4.6Hz, 2H), 0.76 (dd, J=8.0, 3.3Hz, 2H). LC-MS(ESI)m / z 566.15[M+H].

[0794] I-33

[0795]

[0796] Step 1: 2-Bromo-3-isopropoxypyridine

[0797] To a stirred solution of 3-pyridinol, 2-bromo- (1 g, 5.747 mmol, 1 eq) and 2-iodopropane (1.17 g, 6.896 mmol, 1.2 eq) in DMF (10 mL) was added CsCO (3.75 g, 11.494 mmol, 2 eq) at room temperature under an air atmosphere. The resulting mixture was stirred at room temperature for 2 h under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography (C18 silica gel column, eluted with 10% to 100% MeCN / water (0.1% FA) in 10 min with a UV detector (254 nm) to give 2-bromo-3-isopropoxypyridine (540 mg, 43.48%) as a brown solid. LC-MS (ESI) m / z 216 [M+H].

[0798] Step 2: [4-(3-Isopropoxypyridin-2-yl)phenyl]methanol

[0799] To a stirred solution of 2-bromo-3-isopropoxypyridine (540 mg, 2.499 mmol, 1 eq) and 4-(hydroxymethyl)phenylboronic acid (455.72 mg, 2.999 mmol, 1.2 eq) in dioxane (10 mL) was added Pd(dppf)Cl (182.86 mg, 0.250 mmol, 0.1 eq), KPO (1326.18 mg, 6.248 mmol, 2.5 eq) and H0 (2.5 mL) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 90° C. under a nitrogen atmosphere for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluting with 10% to 100% MeCN / water (0.1% FA) over 10 min with UV detection (254 nm) to afford [4-(3-isopropoxypyridin-2-yl)phenyl]methanol (600 mg, 98.68%) as a yellow oil. LC-MS (ESI) m / z 244 [M+H].

[0800] Step 3: 2-[4-(Chloromethyl)phenyl]-3-isopropoxypyridine

[0801] To a stirred solution of [4-(3-isopropoxypyridin-2-yl)phenyl]methanol (500 mg, 2.055 mmol, 1 eq) in DCM (10 mL, 131.088 mmol, 63.79 eq) was added thionyl chloride (2.5 mL) dropwise at 0° C. under an air atmosphere. The resulting mixture was stirred at 0° C. for 2 hours under a nitrogen atmosphere. The residue was purified by reverse-phase flash chromatography (C18 silica gel column, eluted with 10% to 100% MeCN / water (0.1% FA) over 10 min with a UV detector (254 nm) to give 2-[4-(chloromethyl)phenyl]-3-isopropoxypyridine (500 mg, 92.95%) as an off-white solid. LC-MS (ESI) m / z 262 [M+H].

[0802] Step 4: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-{[4-(3-isopropoxypyridin-2-yl)phenyl]methyl}pyrido[2,3-d]pyrimidin-7-one

[0803] To a stirred solution of 2-[4-(chloromethyl)phenyl]-4-(trifluoromethyl)pyrimidine (50 mg, 0.183 mmol, 1 eq) and 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8H-pyrido[2,3-d]pyrimidin-7-one (64.98 mg, 0.220 mmol, 1.2 eq) in DMF (1 mL) was added 1,1,3,3-tetramethylguanidine (31.68 mg, 0.274 mmol, 1.5 eq) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 3 h. The residue was purified by reverse phase flash chromatography (C18 silica gel column, eluting with 10% ...

Claims

1. A compound of formula I: or a pharmaceutically acceptable salt thereof, wherein: is a single bond or a double bond; X 1 is O, N, N(R 11 ), C(O), CR 12 or C(R 12 )2; X 2 is O, N, N(R 11 ), C(O), CR 12 or C(R 12 )2; X 3 N(R 11 ), C(O) or C(R 12 )2; X 4 N or CR 12 ; X 5 N or CR 12 ; Each R 11 are independently hydrogen, deuterium, R or -C(O)-R; Each R 12 are independently hydrogen, deuterium, halogen, R, -OR, -NHR, -N(R)2, -C(O)-R, -COOR, -C(O)-NHR or -C(O)-N(R)2; Ring A is a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S; Each R 1 are independently halogen, R, -OR, -NHR, -N(R)2, -C(O)-R, -C(O)-NHR or -C(O)-N(R)2; Ring B is selected from a benzene ring, a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S, or a 5- to 6-membered heterocyclyl ring having 1 to 4 heteroatoms selected from N, O and S; Each R 2 are independently halogen, R, -OR, -NHR, -N(R)2 or -C(O)-R; R 3 is optionally substituted phenyl, or optionally substituted 5- to 6-membered heteroaryl having 1 to 4 heteroatoms selected from N, O and S; R 4 and R 5 Each of is independently hydrogen, deuterium, halogen, R, -OR, -NHR or -N(R)2; or R 4 and R 5 together with the atoms to which they are attached form an optionally substituted ring selected from a C3-C7 carbocycle or a 3- to 7-membered heterocycle having 1 to 4 heteroatoms selected from N, O and S; Each R is independently an optionally substituted C 1-6 an aliphatic or optionally substituted ring selected from a benzene ring, a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S, a C3-C7 carbocyclic ring, or a 3- to 7-membered heterocyclic ring having 1 to 4 heteroatoms selected from N, O or S; n is 0, 1, 2 or 3; or m is 0, 1, 2 or 3.

2. The compound according to claim 1, wherein the compound has formula II or formula II':

3. The compound according to claim 1, wherein the compound has formula III or formula III':

4. The compound according to claim 1, wherein the compound has Formula IV or Formula IV':

5. The compound according to claim 1, wherein the compound has Formula V or Formula V':

6. The compound according to claim 1, wherein the compound has Formula VI or Formula VI':

7. The compound according to claim 1, wherein the compound has Formula VII or Formula VII':

8. The compound according to claim 1, wherein the compound has Formula VIII or Formula VIII':

9. The compound according to claim 1, wherein the compound has Formula IX or Formula IX':

10. The compound according to claim 1, wherein the compound has formula X or formula X':

11. The compound according to claim 1, wherein the compound has Formula XI or Formula XI':

12. The compound according to claim 1, wherein the compound has Formula XII or Formula XII': Where R 21 is optionally substituted C 1-6 Fatty.

13. The compound according to claim 1, wherein the compound has Formula XIII or Formula XIII': Where R 22 is an optionally substituted ring selected from a benzene ring and a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms selected from N, O and S.

14. The compound according to claim 1, wherein the compound has Formula XIV or Formula XIV': Where R 21 is optionally substituted C 1-6 Fatty.

15. A compound according to any one of claims 1 to 14, wherein each R 11 are independently hydrogen or optionally substituted C 1-6 Fatty.

16. A compound according to any one of claims 1 to 15, wherein each R 12 are independently hydrogen, halogen, optionally substituted C 1-6 Aliphatic, -C(O)-R, -COOR or -C(O)-N(R)2. 17 . The compound according to claim 1 , wherein ring A is a 5- to 6-membered heteroaromatic ring having 1 to 4 heteroatoms N.

18. The compound according to claim 17, wherein ring A is 19. A compound according to any one of claims 1 to 18, wherein each R 1 is independently halogen, R or -OR.

20. The compound according to claim 19, wherein each R 1 are independently -Cl, -F, -OCH3, -OCD3, -O-CHF2, -O-CF3 or 21 . The compound according to claim 1 , wherein ring B is a benzene ring, a 6-membered heteroaromatic ring having 1 to 4 heteroatoms N, or a 6-membered heterocyclyl ring having 1 to 4 heteroatoms N.

22. The compound according to claim 21, wherein ring B is 23. A compound according to any one of claims 1 to 22, wherein each R 2 are independently halogen, R, -OR or -C(O)-R.

24. The compound according to claim 23, wherein each R 2 are independently -F, -CH3, -CD3, -OCH3 or -OCD3.

25. A compound according to any one of claims 1 to 24, wherein R 3 is optionally substituted 26. The compound according to claim 25, wherein R 3 for Each R 13 R is independently halogen, R or -OR, and q is 0, 1, 2 or 3.

27. The compound according to claim 26, wherein R 13 For -F, -CH3, -CD3, -CHF2, -CF3, -OCH3, -OCD3, 28. The compound according to claim 26, wherein R 3 for 29. A compound according to any one of claims 1 to 28, wherein R 4 and R 5 Each of is independently hydrogen, deuterium or an optionally substituted C 1-6 Fatty.

30. A compound selected from the compounds described in Table 1, or a pharmaceutically acceptable salt thereof.

31. A pharmaceutical composition comprising a compound according to any one of claims 1 to 30 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, adjuvant or vehicle.

32. A method for treating a disease or condition associated with ubiquitin-specific processing protease 1 (USP1) in a patient, the method comprising administering to the patient a compound according to any one of claims 1 to 30, or a pharmaceutically acceptable salt thereof.

33. The method of claim 32, wherein the disease or condition is cancer.

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