JP2023553866A - TEAD inhibitors and their uses - Google Patents

Abstract

The present invention provides compounds, compositions thereof, and methods of using them.

Description

SEQUENCE LISTING This application contains a Sequence Listing submitted electronically in ASCII format, which is incorporated herein by reference in its entirety. The above ASCII copy created on November 30, 2021 has the name 187446_SL. txt and has a size of 23,551 bytes.

The present invention relates to compounds and methods useful for inhibiting transcriptional enhancer-associated domains (TEADs). The invention also provides pharmaceutically acceptable compositions comprising the compounds of the invention, as well as methods of using the compositions in the treatment of various diseases, disorders, and conditions described herein. .

Yes-associated protein (YAP) and transcriptional coactivator with PDZ binding motif (TAZ) are transcriptional coactivators of the Hippo pathway network and control cell proliferation, migration, and apoptosis. Inhibition of the Hippo pathway promotes the translocation of YAP/TAZ to the nucleus, where YAP/TAZ interacts with TEAD transcription factors and coactivates target gene expression, promoting cell proliferation. Hyperactivation of YAP and TAZ and/or mutations in one or more elements of the Hippo pathway network have been suggested in a number of cancers.

The Hippo signaling cascade is an important pathway for cancer biogenesis and tumor maintenance. The Hippo pathway is severely mutated across many cancer indications through loss of functional mutations in genes such as NF2. These tumor-promoting mutations result in constitutional activation of the downstream transcriptional coactivators YAP and TAZ, which drive expression of many pro-survival and proliferative genes through essential interactions with TEAD protein family members. bring about. In addition, this unrestricted transcriptional program causes enhanced immunosuppression in the tumor microenvironment. As described herein, to target this oncogenic pathway, we downregulate YAP- and TAZ-dependent transcription by selectively binding to TEAD and impairing the interaction of YAP and TAZ. A novel small molecule inhibitor has been identified. As shown herein, these TEAD inhibitors prevent TEAD palmitoylation, which is essential for YAP and TEAD interaction. Furthermore, the TEAD inhibitors described herein inhibit in vitro growth of YAP-dependent (ie, Hippo pathway-deficient cancer cell lines), but not of Hippo pathway wild-type cancer cell lines. Importantly, as shown herein, the TEAD inhibitor compounds of the invention did not affect the survival of differentiated mouse podocyte cell lines or impair mouse kidney histology. Subsequent in vivo experiments show that the TEAD inhibitors described herein downregulate YAP-dependent genes in human tumor xenografts after oral administration. Additionally, the TEAD inhibitors described herein exhibit single-agent tumor growth inhibition of human tumor xenografts in mice at well-tolerated oral doses. The data described herein demonstrate the ability of the small molecule TEAD inhibitors provided herein to target the Hippo pathway in cancer.

It has now been found that the compounds of the present invention, and pharmaceutically acceptable compositions thereof, are effective as TEAD inhibitors. In one aspect, the invention provides compounds of formula I:
or a pharmaceutically acceptable salt thereof, each variable being independently as defined herein and as described in the embodiments herein.

The compounds of the invention, and their pharmaceutically acceptable salts and compositions, are useful in the treatment of various diseases, disorders, or conditions associated with TEAD. Such diseases, disorders, or conditions include cell proliferative diseases (eg, cancers as described herein).

A schematic diagram of Hippo pathway signaling is shown.

1. Summary of Certain Embodiments of the Invention The compounds of the invention, as well as their pharmaceutical salts and compositions, are useful as inhibitors of TEAD. Without wishing to be bound by any particular theory, the compounds of the present invention, and pharmaceutical compositions thereof, inhibit the activity of TEAD and, therefore, inhibit TEAD-associated diseases and disorders, such as cancer. , or is believed to treat a medical condition.

In one aspect, the invention provides compounds of formula I:
or a pharmaceutically acceptable salt thereof, wherein:
L ¹ is a covalent bond or a C _1-6 divalent straight or branched hydrocarbon chain (where 1, 2, or 3 methylene units of the chain are independently -N(R )-, -O-, or -C(O)-),
Ring A may each be substituted,
selected from
Ring B is
selected from
Each ^RW is independently
selected from
Each R ² is independently -OR, -C(O)NR ₂ , optionally substituted -C _1-6 aliphatic,
selected from
each Y is independently N or ^CR5 ;
each R ³ is independently H or an optionally substituted -C _1-6 aliphatic;
Each R ⁴ is independently -S(O) ₂ NR ₂ , -S(O) ₂ R, -C(O)NR ₂ , -C(O)R, or optionally substituted -C _{1 -6} aliphatic;
Each R ⁵ is independently R, -CN, -C(O)R, -C(O)NR ₂ , or 1 to 2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. an optionally substituted 5- to 6-membered heteroaryl having
each m is independently 0, 1, or 2;
p is 0, 1, or 2;
Each R is independently H, optionally substituted -C _1-6 aliphatic, optionally substituted 3- to 8-membered saturated or partially unsaturated monocyclic carbocyclyl, or nitrogen, oxygen or an optionally substituted 3- to 8-membered saturated or partially unsaturated monocyclic heterocyclyl having 1 to 2 heteroatoms independently selected from sulfur.

2. Compounds and definitions:
Compounds of the invention include those described generally herein and are further illustrated by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise specified. For purposes of the present invention, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, ^75th Ed. Additionally, the general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5 ^t. hEd. , Ed. : Smith, M. B. and March, J. , John Wiley & Sons, New York: 2001, the entire contents of which are incorporated herein by reference.

The term "aliphatic" or "aliphatic group," as used herein, is a fully saturated or one or more unsaturated units; means a straight (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is a monocyclic or bicyclic hydrocarbon containing unsaturated units, but not the remainder of the molecule. is not aromatic (also referred to herein as "carbocyclic,""cycloaliphatic," or "cycloalkyl") having a single point of attachment to. Unless otherwise specified, aliphatic groups contain 1-6 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. In still other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms, and in still other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms. . In some embodiments, "cycloaliphatic" (or "carbocycle" or "cycloalkyl") refers to one or more fully saturated or having a single point of attachment to the rest of the molecule. means a monocyclic C ₃ -C ₆ hydrocarbon containing unsaturated units of, but is not aromatic. Suitable aliphatic groups include straight-chain or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups, and hybrids thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cyclo and (alkyl)alkenyl, but are not limited to these.

As used herein, the term "bicyclic ring" or "bicyclic system" generally refers to a saturated or , means any bicyclic system, ie carbocyclic or heterocyclic system, having one or more units of unsaturation. Thus, the term includes all permissible ring fusions, such as ortho-fused or spirocyclic. As used herein, the term "heterobicyclic" is a subset of "bicyclic" in which one or more heteroatoms must be present in one or both rings of the bicyclic ring. . Such heteroatoms may be present at the linkage of the ring and may be substituted, such as nitrogen (including N-oxides), oxygen, sulfur (including oxidized forms such as sulfones and sulfonates), phosphorus (including oxidized forms such as sulfones and sulfonates), (including oxidized forms such as phosphate), boron, and the like. In some embodiments, bicyclic groups have 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 system, i.e., a carbocyclic or heterocyclic saturated or partially unsaturated system having at least one bridge. means. As defined by IUPAC, a "bridge" is an unbranched chain of atoms, or atoms, or a valence bond that connects two bridgeheads; Any skeletal atom of a ring system that is bonded to a skeletal atom (other than hydrogen). In some embodiments, a bridged bicyclic group has 7 to 12 ring members and 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Such bridged bicyclic groups are well known in the art and include the groups described below, where each group is attached to the remainder of the molecule at any substitutable carbon or nitrogen atom. Unless otherwise specified, bridged bicyclic groups can be substituted with one or more substituents as described for aliphatic groups. Additionally or alternatively, any substitutable nitrogen of the bridged bicyclic group is optionally substituted. Exemplary bicyclic rings include:
Exemplary bridged bicyclics include:

The term "lower alkyl" means a C _1-4 straight or branched alkyl group. Exemplary lower alkyl groups are methyl, ethyl, isopropyl, butyl, isobutyl, and tert-butyl.

The term "lower haloalkyl" means a C _1-4 straight or branched alkyl group substituted with one or more halogen atoms.

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

The term "unsaturated" as used herein means that the moiety has one or more units of unsaturation.

As used herein, the term "divalent C _1-8 (or C _1-6 ) saturated or unsaturated, straight or branched hydrocarbon chain" means a straight chain as defined herein refers to divalent alkylene, alkenylene, and alkynylene chains in the form of divalent or branched forms.

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

The term "alkenylene" refers to a divalent alkenyl group. A substituted alkenylene chain is a polymethylene group containing at least one double bond in which one or more hydrogen atoms have been replaced by a substituent. Suitable substituents include those described below as substituted aliphatic groups.

を有する二価のシクロプロピル基を意味する。 As used in the present invention, the term "cyclopropylenyl" refers to the following structure:

means a divalent cyclopropyl group having

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

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

The terms "heteroaryl" and "heteroar-" used alone or as part of a larger moiety, such as "heteroaralkyl" or "heteroaralkoxy" having 5 to 10 ring atoms, preferably 5, 6 or 9 ring atoms, and having 6, 10 or 14 pi electrons shared in the cyclic arrangement, and in addition to the carbon atoms, means a group having 1 to 5 heteroatoms. The term "heteroatom" means nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of basic nitrogen. Heteroaryl groups include thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, napthyridinyl, and pteridinyl. However, it is not limited to these. As used herein, the terms "heteroaryl" and "heteroar-" include a heteroaromatic ring fused to one or more aryl, cycloaliphatic, or heterocyclyl rings and a radical or Also included are groups in which the point of attachment is on the heteroaromatic ring. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolidinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, Mention may be made of phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. A heteroaryl group may be monocyclic or bicyclic. The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic," all of which are substituted or Contains good rings. The term "heteroaralkyl" refers to an alkyl group substituted with a heteroaryl, wherein the alkyl and heteroaryl moieties may be independently substituted.

As used herein, the terms "heterocycle,""heterocyclyl,""heterocyclicradical," and "heterocyclic ring" are used interchangeably and are either saturated or partially unsaturated. a stable 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic group which, in addition to carbon atoms, has one or more, for example 1 to 4, heteroatoms as defined above. means part. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes substituted nitrogens. By way of example, in a saturated or partially unsaturated ring with 0 to 3 heteroatoms selected from oxygen, sulfur or nitrogen, nitrogen can be N (as in 3,4-dihydro-2H-pyrrolyl), NH ( (as in pyrrolidinyl), or ⁺ NR (as in N-substituted pyrrolidinyl).

A heterocyclic ring may be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure, and any of the ring atoms may be substituted. There is. Examples of such saturated or partially unsaturated heterocyclic radicals are tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl. , piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms "heterocycle", "heterocyclyl", "heterocyclyl ring", "heterocyclic group", "heterocyclyl moiety", and "heterocyclic radical" are used interchangeably herein, and a heterocyclyl ring is Also included are groups fused to one or more aryl, heteroaryl, or alicyclic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl. Heterocyclyl groups may be monocyclic or bicyclic. The term "heterocyclylalkyl" means an alkyl group substituted with heterocyclyl, wherein the alkyl and heterocyclyl moieties may be independently substituted.

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

As described herein, compounds of the invention may contain "optionally substituted" moieties. Generally, the term "substituted", whether or not preceded by the term "optionally", means that one or more hydrogens of the specified moiety are replaced with a suitable substituent. Unless otherwise specified, an "optionally substituted" group can have a suitable substituent at each substitutable position of the group, and in any given structure more than one position is If it can be substituted with more than one substituent selected from a particular group, the substituents may be the same or different at all positions. Combinations of substituents envisioned by this invention are preferably those that result in the formation of stable or chemically suitable compounds. As used herein, the term "stable" refers to the production, detection, and, in certain embodiments, recovery, purification, and use of a compound for one or more of the purposes provided in this disclosure. It relates to compounds that remain substantially unchanged when subjected to conditions that permit their use.

Each optional substituent on a substitutable carbon is halogen; -(CH ₂ ) _0-4 R°; -(CH ₂ ) _0-4 OR°; -O(CH ₂ ) _0-4 R°; -O-(CH ₂ ) _0-4 C(O)OR°; -(CH ₂ ) _0-4 CH(OR°) ₂ ;-(CH ₂ ) _0-4 SR°; Substituted with R° -(CH ₂ ) _0-4 Ph; optionally substituted with R° -(CH ₂ ) _0-4 O(CH ₂ ) _0-1 Ph; optionally substituted with R° -CH =CHPh; optionally substituted with R° -(CH ₂ ) _0-4 O(CH ₂ ) _0-1 -pyridyl; -NO ₂ ; -CN; -N ₃ ; -(CH ₂ ) _0-4 N(R°) ₂ ;-(CH ₂ ) _0-4 N(R°)C(O)R°;-N(R°)C(S)R°;-(CH ₂ ) _0-4 N( R°)C(O)NR° ₂ ;-N(R°)C(S)NR° ₂ ;-(CH ₂ ) _0-4 N(R°)C(O)OR°;-N(R° )N(R°)C(O)R°;-N(R°)N(R°)C(O)NR° ₂ ;-N(R°)N(R°)C(O)OR°; -(CH ₂ ) _0-4 C(O)R°;-C(S)R°;-(CH ₂ ) _0-4 C(O)OR°;-(CH ₂ ) _0-4 C(O) SR°;-(CH ₂ ) _0-4 C(O)OSiR° ₃ ;-(CH ₂ ) _0-4 OC(O)R°;-OC(O)(CH ₂ ) _0-4 SR-, SC (S)SR°;-(CH ₂ ) _0-4 SC(O)R°;-(CH ₂ ) _0-4 C(O)NR° ₂ ;-C(S)NR° ₂ ;-C(S )SR°;-SC(S)SR°,-(CH ₂ ) _0-4 OC(O)NR° ₂ ;-C(O)N(OR°)R°;-C(O)C(O) R°;-C(O)CH ₂ C(O)R°;-C(NOR°)R°;-(CH ₂ ) _0-4 SSR°;-(CH ₂ ) _0-4 S(O) ₂ R°;-(CH ₂ ) _0-4 S(O) ₂ OR°;-(CH ₂ ) _0-4 OS(O) ₂ R°;-S(O) ₂ NR° ₂ ;-S(O) (NR°)R°;-S(O) ₂ N=C(NR° ₂ ) ₂ ;-(CH ₂ ) _0-4 S(O)R°;-N(R°)S(O) ₂ NR ° ₂ ;-N(R°)S(O) ₂ R°;-N(OR°)R°;-C(NH)NR° ₂ ;-P(O) ₂ R°;-P(O)R ° ₂ ;-OP(O)R° ₂ ;-OP(O)(OR°) ₂ ;SiR° ₃ ;-(C _1-4 straight chain or branched alkylene)ON(R°) ₂ ; or a monovalent substituent independently selected from -(C _1-4 straight or branched alkylene)C(O)ON(R° ₂ ).

Each R° is independently hydrogen, C _1-6 aliphatic, -CH ₂ Ph, -O(CH ₂ ) _0-1 Ph, -CH ₂ - (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 , R°, with intervening atom(s), having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 3- to 12-membered may be substituted on the saturated carbon atom of R° by a divalent substituent selected from =O and =S, Alternatively, each R° can be halogen, -(CH ₂ ) _0-2 R ^● , -(halo R ^● ), -(CH ₂ ) _0-2 OH, -(CH ₂ ) _0-2 OR ^● , -( CH ₂ ) _0-2 CH(OR ^● ) ₂ ; -O (halo R ^● ), -CN, -N ₃ , -(CH ₂ ) _0-2 C(O)R ^● , -(CH ₂ ) _{0- 2} C(O)OH, -(CH ₂ ) _0-2 C(O)OR ^● , -(CH ₂ ) _0-2 SR ^● , -(CH ₂ ) _0-2 SH, -(CH ₂ ) _{0- 2} NH ₂ , -(CH ₂ ) _0-2 NHR ^● , -(CH ₂ ) _0-2 NR ^● ₂ , -NO ₂ , -SiR ^● ₃ , -OSiR ^● ₃ , -C(O)SR ^● , - (C _1-4 straight or branched chain alkylene) C(O)OR ^● , or -SSR ^● may be substituted with monovalent substituents independently selected from.

Each R ^● is C _1-4 aliphatic, -CH ₂ Ph, -O(CH ₂ ) _0-1 Ph, or 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. independently selected from 5- to 6-membered saturated, partially unsaturated, or aryl ^rings having Substituted only by halogens or any substituents on saturated carbons are =O, =S, =NNR ^* ₂ , =NNHC(O)R ^* , =NNHC(O)OR ^* , =NNHS( O) ₂ R ^* , =NR ^* , =NOR ^* , -O(C(R ^* ₂ )) _2-3 O-, or -S(C(R ^* ₂ )) _2-3 Independently from S- The divalent substituent bond to the adjacent substitutable carbon in the selected divalent substituent or "optionally substituted" group is -O(CR ^* ₂ ) _2-3 O- and each independent occurrence of R ^* is an unsubstituted group having 0 to 4 heteroatoms independently selected from hydrogen, C _1-6 aliphatic, or nitrogen, oxygen, or sulfur. selected from 5- to 6-membered saturated, partially unsaturated, or aryl rings.

When R ^* is C _1-6 aliphatic, R ^* is halogen, -R ^● , - (halo R ^● ), -OH, -OR ^● , -O (halo R ^● ), -CN, -C May be substituted with (O)OH, -C(O)OR ^● , -NH ₂ , -NHR ^● , -NR ^● ₂ , or -NO ₂ , where each R ^● is a C _1-4 fatty acid a saturated _5- to ₆ -membered ring having ₀ to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; Partially unsaturated or independently selected from aryl rings, each R ^● is unsubstituted or, where preceded by "halo", substituted only by one or more halogens.

Any substituent on the substitutable nitrogen can be independently -R ^† , -NR ^† ₂ , -C(O)R ^† , -C(O)OR ^† , -C(O)C(O) R ^† , -C(O)CH ₂ C(O)R ^† , -S(O) ₂ R ^† , S(O) ₂ NR ^† ₂ , -C(S)NR ^† ₂ , -C(NH)NR ^† ₂ , or -N(R ^† )S(O) ₂ R ^† , where each R ^† is independently hydrogen, C _1-6 aliphatic, unsubstituted -OPh, or nitrogen, oxygen or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from sulfur, or two independent occurrences of R ^† are , unsubstituted 3- to 12-membered saturated, partially unsaturated, with 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, with intervening atom(s); or form an aryl mono- or bicyclic ring, where R ^† is C _1-6 aliphatic, R ^† is halogen, -R ^● , -(halo R ^● ), -OH, -OR Substituted with ^● , -O (halo R ^● ), -CN, -C(O)OH, -C(O)OR ^● , -NH ₂ , -NHR ^● , -NR ^● ₂ , or -NO ₂ Often, where each R ^● is independently selected from C _1-4 aliphatic, -CH ₂ Ph, -O(CH ₂ ) _0-1 Ph, or nitrogen, oxygen, or sulfur. independently selected from 5- to 6-membered saturated, partially unsaturated, or aryl rings having 4 heteroatoms, each R ^● is unsubstituted or where preceded by "halo" , substituted only by one or more halogens.

As used herein, the term "pharmaceutically acceptable salts" means salts that, within the scope of sound medical judgment, can be used in humans and Refers to those salts which are suitable for use in contact with animal tissues and which are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. J. Berge et al., incorporated herein by reference. Pharmaceutical Sciences, 1977, 66, 1-19 describes pharmaceutically acceptable salts in detail. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are inorganic acids such as hydrochloric, hydrobromic, phosphoric, sulfuric and perchloric acids, or acetic, oxalic, maleic, tartaric, citric acids. , with organic acids such as 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. , camphor sulfonate, citrate, cyclopentane propionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate Salt, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonic acid salt, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectate, persulfate, 3-phenylpropionate, Phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Can be mentioned.

Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, and N ⁺ (C _1-4 alkyl) ₄ salts. Exemplary alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium, and amine cations formed with counterions, such as halides, hydroxides, carboxylic acids, etc. salts, sulfates, phosphates, nitrates, lower alkylsulfonates, and arylsulfonates.

Unless otherwise stated, structures depicted herein also refer to all structural isomers (e.g., enantiomers, diastereomers, and geometric (or conformational) isomers), e.g., at each asymmetric center. is intended to include R and S configurations, Z and E double bond isomers, and Z and E conformers. Therefore, in addition to single stereochemical isomers, enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the compounds of the invention are within the scope of the invention. Unless otherwise stated, all tautomeric forms of the compounds of the invention are within the scope of the invention. Additionally, unless otherwise stated, structures depicted herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having this structure that include hydrogen exchange with deuterium or tritium or carbon exchange with ¹³ C or ¹⁴ C enriched carbon are within the scope of the present invention. Such compounds are useful, for example, as analytical tools according to the invention, as probes in biological assays, or as therapeutic agents. In certain embodiments, the warhead portion of the provided compound includes one or more deuterium atoms.

As used herein, the term "inhibitor," or "TEAD inhibitor," or "TEAD antagonist" refers to a compound that binds to and/or inhibits TEAD with measurable affinity. Defined as a compound that inhibits In some embodiments, inhibition in the presence of an 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 less than the signal measured using a negative control under similar conditions. 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 100% lower. The potency of an inhibitor is usually defined by its IC ₅₀ value (half-inhibitory concentration, ie, the concentration required to inhibit 50% of the agonist response). The lower the IC ₅₀ value, the more potent the antagonist and the lower the concentration required to inhibit maximal biological response. In certain embodiments, the inhibitor has an IC ₅₀ and/or binding constant 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. .

As used herein, the terms "measurable affinity" and "measurably inhibits" refer to the relationship between a compound of the invention or a composition thereof and a sample containing a TEAD and a compound or It refers to a measurable change in or inhibition of TEAD activity in the absence of the composition compared to an equivalent sample containing TEAD.

3. Description of exemplary embodiments:
In one aspect, the invention provides compounds of formula I:
or a pharmaceutically acceptable salt thereof, wherein:
L ¹ is a covalent bond or a C _1-6 divalent straight or branched hydrocarbon chain (where 1, 2, or 3 methylene units of the chain are independently -N(R )-, -O-, or -C(O)-),
Ring A may each be substituted,
selected from
Ring B is
selected from
Each ^RW is independently
selected from
Each R ² is independently -OR, -C(O)NR ₂ , optionally substituted -C _1-6 aliphatic,
selected from
each Y is independently N or ^CR5 ;
each R ³ is independently H or an optionally substituted -C _1-6 aliphatic;
Each R ⁴ is independently -S(O) ₂ NR ₂ , -S(O) ₂ R, -C(O)NR ₂ , -C(O)R, or optionally substituted -C _{1 -6} aliphatic;
Each R ⁵ is independently R, -CN, -C(O)R, -C(O)NR ₂ , or 1 to 2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. an optionally substituted 5- to 6-membered heteroaryl having
each m is independently 0, 1, or 2;
p is 0, 1, or 2;
Each R is independently H, optionally substituted -C _1-6 aliphatic, optionally substituted 3- to 8-membered saturated or partially unsaturated monocyclic carbocyclyl, or nitrogen, oxygen or an optionally substituted 3- to 8-membered saturated or partially unsaturated monocyclic heterocyclyl having 1 to 2 heteroatoms independently selected from sulfur.

Generally as defined above, L ¹ is a covalent bond or a C _1-6 divalent straight or branched hydrocarbon chain, where 1, 2, or 3 methylene units of the chain may be independently substituted with -N(R)-, -O-, or -C(O)-.

In some embodiments, L ¹ is a covalent bond.

In some embodiments, L ¹ is a C _1-6 divalent straight or branched hydrocarbon chain, where the 1, 2, or 3 methylene units of the chain are independently: May be substituted with -N(R)-.

In some embodiments, L ¹ is a C _1-6 divalent straight or branched hydrocarbon chain, where the 1, 2, or 3 methylene units of the chain are independently: May be substituted with -O-.

In some embodiments, L ¹ is a C _1-6 divalent straight or branched hydrocarbon chain, where the 1, 2, or 3 methylene units of the chain are independently: May be substituted with -C(O)-.

である。いくつかの実施形態では、Ｌ^１は

である。いくつかの実施形態では、Ｌ^１は

である。いくつかの実施形態では、Ｌ^１は－ＣＨ＝ＣＨ－である。いくつかの実施形態では、Ｌ^１は

である。いくつかの実施形態では、Ｌ^１は

である。いくつかの実施形態では、Ｌ^１は－ＮＨ－Ｃ（Ｏ）－である。 In some embodiments, L ¹ is -NH-. In some embodiments, L ¹ is -NH-CH ₂ -. In some embodiments, L ¹ is -NH-CH ₂ -CH ₂ -. In some embodiments, L ¹ is -CH ₂ -. In some embodiments, L ¹ is
It is. In some embodiments, L ¹ is

It is. In some embodiments, L ¹ is

It is. In some embodiments, L ¹ is

It is. In some embodiments, L ¹ is -CH=CH-. In some embodiments, L ¹ is

It is. In some embodiments, L ¹ is

It is. In some embodiments, L ¹ is -NH-C(O)-.

In some embodiments, L ¹ is selected from those listed in Table 1 below.

から選択される。 As generally defined above, each ring A may be substituted,

selected from.

In some embodiments, ring A is optionally substituted.
It is.

である。 In some embodiments, ring A is optionally substituted.

It is.

である。 In some embodiments, ring A is optionally substituted.

It is.

である。 In some embodiments, ring A is optionally substituted.

It is.

である。 In some embodiments, ring A is optionally substituted.

It is.

である。 In some embodiments, ring A is optionally substituted.

It is.

である。 In some embodiments, ring A is optionally substituted.

It is.

である。 In some embodiments, ring A is optionally substituted.

It is.

である。 In some embodiments, ring A is optionally substituted.

It is.

である。 In some embodiments, ring A is optionally substituted.

It is.

である。 In some embodiments, ring A is optionally substituted.

It is.

である。 In some embodiments, ring A is optionally substituted.

It is.

In some embodiments, ring A is
wherein each R ¹ is independently R, halogen, -CN, -C(O)R, -C(O)NR ₂ , -OR, -SR, -S(O) ₂ NR ₂ , or -S(O) ₂ R, and each n is independently 0, 1, 2, or 3, and each R is independently as defined herein and As described in the Embodiments section of the book.

In some embodiments, R ¹ is R. In some embodiments, R ¹ is halogen. In some embodiments, R ¹ is -CN. In some embodiments, R ¹ is -C(O)R. In some embodiments, R ¹ is -C(O)NR ₂ . In some embodiments, R ¹ is -OR. In some embodiments, R ¹ is -SR. In some embodiments, R ¹ is -S(O) ₂ NR ₂ . In some embodiments, R ¹ is -S(O) ₂ R.

In some embodiments, each R ¹ is independently H, halogen, -C _1-6 aliphatic, optionally substituted with 1 to 6 halogens, a 3- to 8-membered saturated or partially unsaturated monocyclic carbocyclyl, or 1 to 2 independently selected from nitrogen, oxygen, or sulfur optionally substituted with 1 to 6 halogens; A 3- to 8-membered saturated or partially unsaturated monocyclic heterocyclyl having .

In some embodiments, each R ¹ is independently H, -CF ₃ , -C(O)NH ₂ , -CH ₃ , -CH ₂ CH ₃ , -OCH ₃ , -CHF ₂ , -OCF ₃ , -OCHF ₂ , -SCF ₃ , -Cl, -S(O) ₂ -NH ₂ , -OCH ₂ CH ₃ , -F, -C(O)NHCH ₃ , -CN, -S(O) ₂ -CH ₃ , -OCH(CH ₃ ) ₂ , -CH(CH ₃ ) ₂ , -C(CH ₃ ) ₃ , -CH ₂ OH,
It is.

In some embodiments, each R ¹ is independently selected from those listed in Table 1 below.

In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

In some embodiments, ring A is
wherein each of R ¹ , both alone and in combination, is as defined above and described in the embodiments herein.

から選択され、式中、Ｒ^１はそれぞれ、単独及び組み合わせでの両方で、上記で定義され、及び、本明細書内の実施形態に記載されているとおりである。 In some embodiments, ring A is

wherein each R ¹ , both alone and in combination, is as defined above and described in the embodiments herein.

である。 In some embodiments, ring A is

It is.

In some embodiments, ring A is
selected from.

In some embodiments, Ring A is selected from those listed in Table 1 below.

Generally, as defined above, ring B is
wherein R ² , R ³ , R ^w , p, and R ⁴ , both alone and in combination, are as defined above and described in the embodiments herein. That's right.

であり、式中、Ｒ^２及びＲ^ｗのそれぞれは、単独及び組み合わせでの両方で、上記で定義され、及び、本明細書内の実施形態に記載されているとおりである。 In some embodiments, ring B is

and where each of R ² and R ^w , both alone and in combination, is as defined above and described in the embodiments herein.

であり、式中、Ｒ^４及びＲ^ｗのそれぞれは、単独及び組み合わせでの両方で、上記で定義され、及び、本明細書内の実施形態に記載されているとおりである。 In some embodiments, ring B is

where each of R ⁴ and R ^w , both alone and in combination, is as defined above and as described in the embodiments herein.

であり、式中、Ｒ^２及びＲ^ｗのそれぞれは、単独及び組み合わせでの両方で、上記で定義され、及び、本明細書内の実施形態に記載されているとおりである。 In some embodiments, ring B is

and where each of R ² and R ^w , both alone and in combination, is as defined above and described in the embodiments herein.

であり、式中、Ｒ^２及びＲ^ｗのそれぞれは、単独及び組み合わせでの両方で、上記で定義され、及び、本明細書内の実施形態に記載されているとおりである。 In some embodiments, ring B is

and where each of R ² and R ^w , both alone and in combination, is as defined above and described in the embodiments herein.

であり、式中、Ｒ^４及びＲ^ｗのそれぞれは、単独及び組み合わせでの両方で、上記で定義され、及び、本明細書内の実施形態に記載されているとおりである。 In some embodiments, ring B is

where each of R ⁴ and R ^w , both alone and in combination, is as defined above and as described in the embodiments herein.

であり、式中、Ｒ^２及びＲ^ｗのそれぞれは、単独及び組み合わせでの両方で、上記で定義され、及び、本明細書内の実施形態に記載されているとおりである。 In some embodiments, ring B is

and where each of R ² and R ^w , both alone and in combination, is as defined above and described in the embodiments herein.

In some embodiments, ring B is
and where each of R ³ and p, both alone and in combination, are as defined above and as described in the embodiments herein. In some embodiments, ring B is
It is.

であり、式中、Ｒ^ｗは、本明細書で定義され、及び、本明細書の実施形態で記載されているとおりである。いくつかの実施形態では、環Ｂは

であり、式中、Ｒ^ｗは、本明細書で定義され、及び、本明細書の実施形態で記載されているとおりである。 In some embodiments, ring B is

and where R ^w is as defined herein and described in the embodiments herein. In some embodiments, ring B is

and where R ^w is as defined herein and described in the embodiments herein.

In some embodiments, Ring B is selected from those listed in Table 1 below.

から選択される。 Generally, as defined above, R ^w is

selected from.

である。一部の実施形態では、Ｒ^ｗは

である。一部の実施形態では、Ｒ^ｗは

である。一部の実施形態では、Ｒ^ｗは

である。一部の実施形態では、Ｒ^ｗは

である。一部の実施形態では、Ｒ^ｗは

である。一部の実施形態では、Ｒ^ｗは

である。 In some embodiments, R ^w is

It is. In some embodiments, R ^w is

It is. In some embodiments, R ^w is

It is. In some embodiments, R ^w is

It is. In some embodiments, R ^w is

It is. In some embodiments, R ^w is

It is. In some embodiments, R ^w is

It is.

In some embodiments, R ^w is selected from those listed in Table 1 below.

As generally defined above, each R ² is independently -OR, -C(O)NR ₂ , optionally substituted -C _1-6 aliphatic,
wherein each of Y, m, and R ⁵ , both alone and in combination, are as defined above and as described in the embodiments herein.

である。いくつかの実施形態では、Ｒ^２は

である。いくつかの実施形態では、Ｒ^２は

である。いくつかの実施形態では、Ｒ^２は

である。いくつかの実施形態では、Ｒ^２は

である。いくつかの実施形態では、Ｒ^２は

である。いくつかの実施形態では、Ｒ^２は
である。 In some embodiments, R ² is -OR. In some embodiments, R ² is -C(O)NR ₂ . In some embodiments, R ² is an optionally substituted -C _1-6 aliphatic. In some embodiments, R ² is

It is. In some embodiments, R ² is

It is. In some embodiments, R ² is

It is. In some embodiments, R ² is

It is. In some embodiments, R ² is

It is. In some embodiments, R ² is

It is. In some embodiments, R ² is
It is.

である。いくつかの実施形態では、Ｒ^２は
である。いくつかの実施形態では、Ｒ^２は

である。いくつかの実施形態では、Ｒ^２は
である。いくつかの実施形態では、Ｒ^２は

である。いくつかの実施形態では、Ｒ^２は

である。いくつかの実施形態では、Ｒ^２は

である。いくつかの実施形態では、Ｒ^２は

である。いくつかの実施形態では、Ｒ^２は

である。いくつかの実施形態では、Ｒ^２は

である。いくつかの実施形態では、Ｒ^２は

である。いくつかの実施形態では、Ｒ^２は

である。いくつかの実施形態では、Ｒ^２は
である。いくつかの実施形態では、Ｒ^２は

である。 In some embodiments, R ² is
It is. In some embodiments, R ² is

It is. In some embodiments, R ² is
It is. In some embodiments, R ² is

It is. In some embodiments, R ² is
It is. In some embodiments, R ² is

It is. In some embodiments, R ² is

It is. In some embodiments, R ² is

It is. In some embodiments, R ² is

It is. In some embodiments, R ² is

It is. In some embodiments, R ² is

It is. In some embodiments, R ² is

It is. In some embodiments, R ² is

It is. In some embodiments, R ² is
It is. In some embodiments, R ² is

It is.

から選択される。 In some embodiments, R ² is

selected from.

In some embodiments, R ² is
and -OCH ₃ .

In some embodiments, R ² is selected from those listed in Table 1 below.

Each Y is independently N or ^CR5 , generally as defined above.

In some embodiments, Y is N. In some embodiments, Y is ^CR5 . In some embodiments, Y is CH.

In some embodiments, both Ys are N. In some embodiments, both Ys are ^CR5 . In some embodiments, one Y is N and the other Y is ^CR5 . In some embodiments, both Ys are CH. In some embodiments, one Y is N and the other Y is CH.

In some embodiments, Y is selected from those listed in Table 1 below.

Generally as defined above, each R ³ is independently H, -C(O)R, or an optionally substituted -C _1-6 aliphatic, where R is and as described in the embodiments herein.

In some embodiments, ^R3 is H.

In some embodiments, R ³ is -C(O)R.

In some embodiments, R ³ is an optionally substituted -C _1-6 aliphatic.

から選択される。 In some embodiments, R ³ is H, -CH ₃ , -CH ₂ CH ₃ , -C(O)CH ₃ , and

selected from.

In some embodiments, R ³ is selected from those listed in Table 1 below.

As generally defined above, each R ⁴ is independently -S(O) ₂ NR ₂ , -S(O) ₂ R, -C(O)NR ₂ , -C(O)R, or optionally substituted -C _1-6 aliphatic, where each R is independently as defined herein and as described in the embodiments herein.

In some embodiments, R ⁴ is -S(O) ₂ NR ₂ .

In some embodiments, R ⁴ is -S(O) ₂ R.

In some embodiments, R ⁴ is -C(O)NR ₂ .

In some embodiments, R ⁴ is -C(O)R.

In some embodiments, R ⁴ is an optionally substituted -C _1-6 aliphatic.

から選択される。 In some embodiments, R ⁴ is

selected from.

In some embodiments, R ⁴ is
selected from.

In some embodiments, R ⁴ is selected from those listed in Table 1 below.

Each R ⁵ is independently selected from R, -CN, -C(O)R, -C(O)NR ₂ , or nitrogen, oxygen, or sulfur, generally as defined above. an optionally substituted 5- to 6-membered heteroaryl having 1 to 2 heteroatoms, wherein each R is independently as defined herein and as practiced herein. As stated in the form.

In some embodiments, R ⁵ is R.

In some embodiments, R ⁵ is -CN-.

In some embodiments, R ⁵ is -C(O)R.

In some embodiments, R ⁵ is -C(O)NR ₂ .

In some embodiments, R ⁵ is an optionally substituted 5-6 membered heteroaryl having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

In some embodiments, each R ⁵ is H, -CH ₃ , -CD ₃ ,
independently selected from.

In some embodiments, each R ⁵ is -CH ₃ , -CH ₂ CH ₂ OCH ₃ , -CH ₂ CF ₃ , -CH ₂ CH ₂ Cl,
independently selected from.

In some embodiments, R ⁵ is selected from those listed in Table 1 below.

Each m is independently 0, 1, or 2, generally as defined above.

In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2.

In some embodiments, m is selected from those listed in Table 1 below.

Generally, p is 0, 1, or 2 as defined above.

In some embodiments, p is 0. In some embodiments, p is 1. In some embodiments, p is 2.

In some embodiments, p is selected from those listed in Table 1 below.

Generally as defined above, each R is independently H, an optionally substituted -C _1-6 aliphatic, an optionally substituted 3- to 8-membered saturated or partially unsaturated monocyclic ring. carbocyclyl or an optionally substituted 3- to 8-membered saturated or partially unsaturated monocyclic heterocyclyl having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, or sulfur It is.

In some embodiments, R is H.

In some embodiments, R is an optionally substituted -C _1-6 aliphatic. In some embodiments, R is unsubstituted -C _1-6 aliphatic. In some embodiments, R is -C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with -halogen, -CN, or -NO ₂ . In some embodiments, R is -C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with -F. In some embodiments, R is -C _1-3 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with -F. In some embodiments, R is _-CH3 . In some embodiments, R is -CH ₂ CH ₃ . In some embodiments, R is _-CF3 . In some embodiments, R is _-CHF2 .

In some embodiments, R is an optionally substituted 3-8 membered saturated or partially unsaturated monocyclic carbocyclyl. In some embodiments, R is an unsubstituted 3-, 4-, 5-, 6-, 7-, or 8-membered, saturated or partially unsaturated monocyclic carbocyclyl. In some embodiments, R is 3, 4, substituted 1, 2, 3, 4, 5, or 6 times with -halogen, -CN, -NO ₂ , or -C _1-6 aliphatic. A 5-, 6-, 7-, or 8-membered saturated or partially unsaturated monocyclic carbocyclyl, in which the -C _1-6 aliphatic is replaced by -halogen, -CN, or -NO ₂ . , 2, 3, 4, 5, or 6 times. In some embodiments, R is a 3-, 4-, 5-, 6-, 7-, or 8-membered saturated or partially ununion substituted 1, 2, 3, 4, 5, or 6 times with -halogen. is a saturated monocyclic carbocyclyl; in some embodiments, R is 3, 4, 5, substituted 1, 2, 3, 4, 5, or 6 times with -C _1-6 aliphatic; a 6-, 7-, or 8-membered saturated or partially unsaturated monocyclic carbocyclyl, where -C _1-6 aliphatic is 1, 2, 3, 4, 5, or May be substituted 6 times. In some embodiments, R is a 3, 4, 5, 6, 7, or 8-membered saturated or partially unsaturated group substituted 1, 2, 3, 4, 5, or 6 times with -F. is a saturated monocyclic carbocyclyl; in some embodiments, R is 3, 4, 5, substituted 1, 2, 3, 4, 5, or 6 times with -C _1-6 aliphatic; a 6-, 7-, or 8-membered saturated or partially unsaturated monocyclic carbocyclyl, where -C _1-6 aliphatic is 1, 2, 3, 4, 5, or May be substituted 6 times.

In some embodiments, R is an optionally substituted 3- to 8-membered saturated or partially It is an unsaturated monocyclic heterocyclyl. In some embodiments, R is an unsubstituted 3-, 4-, 5-, 6-, 7-, or 8-membered group having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. , a saturated or partially unsaturated monocyclic heterocyclyl. In some embodiments, R is nitrogen, oxygen, substituted 1, 2, 3, 4, 5, or 6 times with -halogen, -CN, -NO ₂ , or -C _1-6 aliphatic, or a 3, 4, 5, 6, 7, or 8-membered saturated or partially unsaturated monocyclic heterocyclyl having 1 to 2 heteroatoms independently selected from sulfur; In, the -C _1-6 aliphatic may be substituted 1, 2, 3, 4, 5, or 6 times with -halogen, -CN, or -NO ₂ . In some embodiments, R is 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, substituted 1, 2, 3, 4, 5, or 6 times with -halogen. is a 3-, 4-, 5-, 6-, 7-, or 8-membered saturated or partially unsaturated monocyclic heterocyclyl having In some embodiments, R is 1-6 independently selected from nitrogen, oxygen, or sulfur, substituted 1, 2, 3, 4, 5, or 6 times with -C _1-6 aliphatic. a 3, 4, 5, 6, 7, or 8-membered saturated or partially unsaturated monocyclic heterocyclyl having 2 heteroatoms, where -C _1-6 aliphatic is - may be substituted 1, 2, 3, 4, 5 or 6 times by halogen; In some embodiments, R is 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, substituted 1, 2, 3, 4, 5, or 6 times with -F. is a 3-, 4-, 5-, 6-, 7-, or 8-membered saturated or partially unsaturated monocyclic heterocyclyl having In some embodiments, R is 1-6 independently selected from nitrogen, oxygen, or sulfur, substituted 1, 2, 3, 4, 5, or 6 times with -C _1-6 aliphatic. a 3, 4, 5, 6, 7, or 8-membered saturated or partially unsaturated monocyclic heterocyclyl having 2 heteroatoms, where -C _1-6 aliphatic is -F may be substituted 1, 2, 3, 4, 5, or 6 times.

In some embodiments, R is
selected from.

In some embodiments, R is selected from those listed in Table 1 below.

In some embodiments, the invention provides compounds of the formula:
or a pharmaceutically acceptable salt thereof, wherein each of R ¹ , L ¹ , R ^w , Y, m, n, and R ⁵ , both alone and in combination, is as defined above. and as described in the embodiments herein.

In some embodiments, the invention provides compounds of the formula:
or a pharmaceutically acceptable salt thereof, wherein each of R ¹ , L ¹ , R ^w , Y, n, and R ⁵ , both alone and in combination, are as defined above; and as described in the embodiments herein.

In some embodiments, the invention provides compounds of the formula:
or a pharmaceutically acceptable salt thereof, wherein each of R ¹ , L ¹ , R ^w , n, and R ⁵ , both alone and in combination, are defined above, and As described in the embodiments herein.

In some embodiments, the invention provides compounds of the formula:
or a pharmaceutically acceptable salt thereof, wherein each of R ¹ , R ^w , L ¹ , and R ⁵ , both alone and in combination, is defined above and as defined herein. As described in the embodiments in the book.

またはこれらの薬学的に許容される塩を提供し、式中、Ｒ、Ｒ^１、Ｌ^１、Ｒ^ｗ、及びｎのそれぞれは、単独及び組み合わせでの両方で、上記で定義され、及び、本明細書内の実施形態に記載されているとおりである。 In some embodiments, the invention provides compounds of the formula:

or a pharmaceutically acceptable salt thereof, wherein each of R, R ¹ , L ¹ , R ^w , and n, both alone and in combination, are defined above, and As described in the embodiments in the specification.

In some embodiments, the invention provides compounds of the formula:
or a pharmaceutically acceptable salt thereof, wherein each of R, R ¹ , L ¹ , R ^w , and n, both alone and in combination, are defined above, and As described in the embodiments in the specification.

In some embodiments, the invention provides compounds of the formula:
or a pharmaceutically acceptable salt thereof, wherein each of R ¹ , L ¹ , R ^w , and n, both alone and in combination, are defined above and as defined herein. As described in the embodiments within.

In some embodiments, the invention provides compounds of the formula:
or a pharmaceutically acceptable salt thereof, wherein each of R ¹ , L ¹ , R ^w , Y, m, n, and R ⁵ , both alone and in combination, is as defined above. and as described in the embodiments herein.

In some embodiments, the invention provides compounds of the formula:
or a pharmaceutically acceptable salt thereof, wherein each of R ¹ , L ¹ , R ^w , Y, n, and R ⁵ , both alone and in combination, are as defined above; and as described in the embodiments herein.

In some embodiments, the invention provides compounds of the formula:
or a pharmaceutically acceptable salt thereof, wherein each of R ¹ , L ¹ , R ^w , n, and R ⁵ , both alone and in combination, are defined above, and As described in the embodiments herein.

またはこれらの薬学的に許容される塩を提供し、式中、Ｒ^１、Ｌ^１、Ｒ^ｗ、及びＲ^５のそれぞれは、単独及び組み合わせでの両方で、上記で定義され、及び、本明細書内の実施形態に記載されているとおりである。 In some embodiments, the invention provides compounds of the formula:

or a pharmaceutically acceptable salt thereof, wherein each of R ¹ , L ¹ , R ^w , and R ⁵ , both alone and in combination, are defined above and as defined herein. As described in the embodiments in the book.

Without wishing to be bound by any particular theory, the compounds provided herein have the advantage that amino acid residues (cysteine, lysine, histidine, or covalently modified (e.g., other residues that can be used), thereby irreversibly inhibiting the protein. In some embodiments, the warhead group can be covalently attached to cysteine. In some embodiments, the warhead group can be covalently attached to serine. In some embodiments, the warhead group can be covalently attached to lysine. In some embodiments, the warhead group can be covalently attached to Cys359 of hTEAD1, Cys405 of hTEAD1, Cys380 of hTEAD2, Cys368 of hTEAD3, and/or Cys367 of hTEAD4. In some embodiments, the warhead group can be covalently attached to Ser356 of hTEAD1, Ser345 and/or Ser377 of hTEAD2, Ser365 of hTEAD3, and/or Ser364 of hTEAD4. In some embodiments, the warhead group can be covalently attached to Lys336 of hTEAD1, Lys357 of hTEAD2, Lys345 of hTEAD3, and/or Lys344 of hTEAD4. Exemplary reference amino acid sequences for human TEAD1, human TEAD2, human TEAD3, and human TEAD4 include UniProt KB ID P28347-1 (SEQ ID NO: 1), UniProtKB ID Q15562 (SEQ ID NO: 2), and UniProtKB ID Q99594 (SEQ ID NO: 2), respectively. SEQ ID NO: 3), and UniProtKB ID Q15561 (SEQ ID NO: 4). The following is published in "Targeting Transcriptional Enhanced Associate Domains (TEADs)," J. D. Med. Chem. This is a sequence alignment of the TEAD coactivator binding domain shown in Table 1 of 2018, 61, 5057-5072.

Exemplary compounds of the invention are listed in Table 1 below.
Table 1: Exemplary compounds

In some embodiments, the invention provides a compound set forth in Table 1 above, or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of the invention is not a compound selected from:

Compounds of the invention are generally prepared or isolated by synthetic and/or semi-synthetic methods known to those skilled in the art for similar compounds, as well as by methods detailed in the Examples herein. It is possible to do so. In some embodiments, the invention provides intermediate compounds described in the Examples, or salts thereof.

4. Use, formulation, and administration:
Pharmaceutically Acceptable Compositions According to another embodiment, the invention comprises a compound of the invention or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant, or vehicle. A pharmaceutical composition is provided. The amount of compound in the compositions of the invention is an amount effective to measurably inhibit TEAD, or a variant or variant thereof, in a biological sample or patient. In certain embodiments, the amount of compound in the compositions of the invention is an amount effective to measurably inhibit TEAD, or a variant or variant thereof, in a biological sample or patient. In certain embodiments, compositions of the invention are formulated for administration to a patient in need thereof. In some embodiments, compositions of the invention are formulated for oral administration to a patient.

The term "patient" or "subject" as used herein means an animal, preferably a mammal, and most preferably a human.

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

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

As used herein, the term "inhibitoryly active metabolite or residue thereof" means that the metabolite or residue thereof is also an inhibitor of TEAD, or a variant or variant thereof.

The compositions of the invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, bucally, vaginally, or via an implanted reservoir. be able to. As used herein, the term "parenteral" includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. is included. Preferably, the composition is administered orally, intraperitoneally or intravenously. Sterile injectable forms of the compositions of this invention may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. can. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium.

For this purpose, any bland fixed oil may be employed including mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives, are useful in the preparation of injectables, as are natural pharmaceutically acceptable oils, such as olive oil or castor oil, especially their polyoxyethylated versions. These oils or suspensions may also contain long-chain alcohol diluents or dispersants commonly used in the formulation of pharmaceutically acceptable dosage forms, including emulsions and suspensions, such as carboxymethyl cellulose or Similar dispersants may be included. Other commonly used surfactants commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms, such as Tween®, Span® , and other emulsifying agents or bioavailability enhancers can also be used for formulation purposes.

The pharmaceutically acceptable compositions of the present invention may be administered orally 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, commonly used carriers include lactose and cornstarch. Lubricating agents such as magnesium stearate are also commonly added. For oral administration in capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring, or coloring agents may also be added.

Alternatively, the pharmaceutically acceptable compositions of the invention can be administered in the form of suppositories for rectal administration. These may be prepared by mixing the drug with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature, thereby dissolving in the rectum and releasing the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

The pharmaceutically acceptable compositions of the invention may also be administered topically, particularly when the target of treatment involves areas or organs readily accessible by topical application, including diseases of the eye, skin, or lower intestinal tract. can do. Suitable topical formulations are readily prepared for each of these areas or organs.

Topical application for the lower intestinal tract may be accomplished with a rectal suppository formulation (see above) or a suitable enema formulation. Topical transdermal patches may also be used.

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

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

Pharmaceutically acceptable compositions of the invention can also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the pharmaceutical formulation art and include benzyl alcohol or other suitable preservatives, absorption enhancers to increase bioavailability, fluorocarbons, and/or other suitable preservatives. It can be prepared as a solution in saline using conventional solubilizing or dispersing agents.

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

The amount of a compound of the invention that may be combined with the carrier materials to produce a composition in a single dosage form will vary depending on the host treated, the particular mode of administration. Preferably, the compositions provided should be formulated such that doses of 0.01 to 100 mg/kg body weight/day of the inhibitor can be administered to patients receiving these compositions.

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

Uses of Compounds and Pharmaceutically Acceptable Compositions Hippo Signaling Network The Hippo signaling network (also known as the Salvador/Warts/Hippo (SWH) pathway) is a master of cell proliferation, death, and differentiation. It is a controlling factor. In some embodiments, the primary function of the Hippo signaling pathway is a transcriptional coactivator, Yes-associated protein (YAP), and its paralog, transcriptional coactivator with PDZ-binding motif (TAZ; (also known as WWTR1). The Hippo kinase cascade phosphorylates and inhibits YAP/TAZ by promoting its cytoplasmic retention and degradation, thereby inhibiting the pro-proliferative functions regulated under the control of YAP/TAZ. In the unphosphorylated/dephosphorylated state, YAP, also known as YAP1 or YAP65, is transported together with TAZ to the nucleus, where YAP and TAZ interact with the TEAD family of transcription factors and are involved in proliferation and It upregulates genes that promote migration and inhibits apoptosis. In some cases, unregulated upregulation of these genes involved in proliferation, migration, and anti-apoptosis leads to the development of cancer. In some cases, YAP/TAZ overexpression is associated with cancer.

Additional core elements of the Hippo signaling pathway include the serine/threonine kinases MST1/2 (homolog of Hippo/Hpo in Drosophila) and Lats1/2 (homolog of Warts/Wts), and their adapter proteins, respectively. Examples include certain Sav1 (homolog of Salvador/Sav) and Mob (MOBKL1A and MOBKL1B; homolog of Mats). Generally, MST1/2 kinase forms a complex with the scaffold protein Sav1, which phosphorylates and activates Lats1/2 kinase. Lats1/2 is also activated by the scaffold protein Mob. Activated Lats1/2 then phosphorylates YAP or its paralog TAZ, rendering it inactive. Phosphorylation of YAP/TAZ results in their nuclear export, retention in the cytoplasm, and degradation by the ubiquitin proteasome system.

In some cases, Lats1/2 phosphorylates YAP at the [HXRXXS] (SEQ ID NO: 9) consensus motif. YAP contains five [HXRXXS] (SEQ ID NO: 9) consensus motifs, where X represents any amino acid residue. In some cases, Lats1/2 phosphorylates YAP at one or more consensus motifs. In some cases, Lats1/2 phosphorylates YAP at all five consensus motifs. In some cases, Lats1/2 phosphorylates at the S127 amino acid position. Phosphorylation of YAP S127 promotes 14-3-3 protein binding and results in cytoplasmic sequestration of YAP. Mutation of YAP at position S127 therefore prevents interaction with 14-3-3 and subsequently promotes nuclear import.

Additional phosphorylation occurs at amino acid position S381 of YAP. Phosphorylation of YAP at position S381 and the corresponding position in TAZ primes both proteins for further phosphorylation events by CK1δ/ε within the degradation motif, which in turn primes the β-TRCP E3 ubiquitin ligase. signal transduction due to interaction with YAP, resulting in polyubiquitination and degradation of YAP.

In some cases, Lats1/2 phosphorylates TAZ at the [HXRXXS] (SEQ ID NO: 9) consensus motif. TAZ contains four [HXRXXS] (SEQ ID NO: 9) consensus motifs, where X represents any amino acid residue. In some cases, Lats1/2 phosphorylates TAZ at one or more consensus motifs. In some cases, Lats1/2 phosphorylates TAZ at all four consensus motifs. In some cases, Lats1/2 phosphorylates at the S89 amino acid position. Phosphorylation of TAZ S89 promotes 14-3-3 protein binding and results in cytoplasmic sequestration of TAZ. Mutation of TAZ at position S89 therefore prevents interaction with 14-3-3 and subsequently promotes nuclear import.

In some embodiments, phosphorylated YAP/TAZ accumulates in the cytoplasm and undergoes SCF ^β-TRCP -mediated ubiquitination and subsequent proteasomal degradation. In some cases, the Skp, Cullin, F-box-containing complex (SCF complex) is a multi-protein E3 ubiquitin ligase complex that includes F-box family member proteins (e.g., Cdc4), Skp1, cross-linking proteins, and RBX1. , which contains a small RING finger domain that interacts with E2-ubiquitin conjugating enzymes. In some cases, the F-box family includes more than 40 members, with exemplary members including F-box/WD repeat-containing protein 1A (FBXW1A, βTrCP1, Fbxwl, hsSlimb, plkappaBalpha-E3 receptor subunit ), and S-phase kinase-related protein 2 (SKP2). In some embodiments, the SCF complex (eg, SCF ^βTrCP1 ) interacts with an E1 ubiquitin activating enzyme and an E2 ubiquitin conjugating enzyme to catalyze the transfer of ubiquitin to a YAP/TAZ substrate. Exemplary E1 ubiquitin activating enzymes include those encoded by the following genes: UBA1, UBA2, UBA3, UBA5, UBA5, UBA7, ATG7, NAEl, and SAE1. Exemplary E2 ubiquitin conjugating enzymes include the following genes: UBE2A, UBE2B, UBE2C, UBE2D1, UBE2D2, UBE2D3, UBE2E1, UBE2E2, UBE2E3, UBE2F, UBE2G1, UBE2G2, UBE2H, UBE2I, UBE2J1, UBE2J2, UBE2K, UBE2L3, These include those coded by UBE2L6, UBE2M, UBE2N, UBE20, UBE2Q1, UBE2Q2, UBE2R1, UBE2R2, UBE2S, UBE2T, UBE2U, UBE2V1, UBE2V2, UBE2Z, ATG2, BIRC5, and UFC1. In some embodiments, the ubiquitinated YAP/TAZ is further subjected to a 26S proteasome-mediated degradation process.

In some embodiments, the Hippo pathway is upstream regulated by several different families of regulators. In some cases, the Hippo pathway is controlled by G proteins and their coupled receptors, the Crumbs complex, upstream regulators of MSt kinase, and adherens junctions.

Interaction of YAP/TAZ with TEAD In some embodiments, unphosphorylated and/or dephosphorylated YAP/TAZ accumulates in the nucleus. In the nucleus, YAP/TAZ binds to transcription factors of the TEAD family (e.g., human TEAD1 (UniProt KB ID P28347-1 (SEQ ID NO: 1)), human TEAD2 (UniProtKB ID Q15562 (SEQ ID NO: 2)), human TEAD3 (UniProtKB ID Q99594 (SEQ ID NO: 3)) and human TEAD4 (UniProtKB ID Q15561 (SEQ ID NO: 4)) and activates genes involved in anti-apoptosis and proliferation, such as CTFG, Cyr61, and FGF1. do.

Proteomic and biochemical studies have shown that TEAD (TEA domain) transcription factors are palmitoylated at evolutionarily conserved cysteine residues. We discovered three cysteine residues (C53S, C327S, and C359S) that are conserved during evolution and mutated to serine in human TEAD1 and tested whether the mutations affect palmitoylation of TEAD1. The C359S mutant showed the greatest loss of palmitoylation, and C327S and C53S also showed reduced palmitoylation. These results suggest that C359 plays an important role in palmitoylation of TEAD1. Furthermore, the combined mutation C53/327/359S (3CS) of all three cysteine residues completely abolished TEAD1 palmitoylation, suggesting that these residues are involved in TEAD1 palmitoylation. ing. TEAD has been found to undergo PAT-independent self-palmitoylation under physiological concentrations of palmitoyl 1-CoA. Furthermore, autopalmitoylation plays an important role in regulating TEAD-YAP association and their physiological functions in vitro and in vivo. Chan, et al. Nature Chem. Biol. 12, pages 282-289 (2016); Noland, et al. Structure, 24, 1-8 (2016); Gibault et al. J. Med. Chem. 61, 5057-5072 (2018). Therefore, palmitoylation of TEAD plays an important role in the regulation of the Hippo pathway transcription complex.

In some embodiments, compounds disclosed herein modulate the interaction of YAP/TAZ and TEAD. In some embodiments, the compounds disclosed herein bind to TEAD, YAP, or TAZ and prevent the interaction of YAP/TAZ and TEAD.

In some embodiments, the compounds described herein irreversibly inhibit the TEAD transcription factor. In some embodiments, the transcription factor is TEAD1. In some embodiments, the transcription factor is TEAD2. In some embodiments, the transcription factor is TEAD3. In some embodiments, the transcription factor is TEAD4. In some embodiments, a compound described herein covalently binds to a TEAD transcription factor (eg, TEAD1, TEAD2, TEAD3, or TEAD4). In some embodiments, the compounds described herein irreversibly inhibit the activity of a TEAD transcription factor (eg, TEAD1, TEAD2, TEAD3, or TEAD4). In some embodiments, the compounds described herein covalently inhibit the activity of a TEAD transcription factor (eg, TEAD1, TEAD2, TEAD3, and/or TEAD4).

In some embodiments, the compounds disclosed herein bind to TEAD1 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD2 and prevent or inhibit the interaction of YAP and TEAD2. In some embodiments, the compounds disclosed herein bind to TEAD3 and prevent or inhibit the interaction of YAP and TEAD3. In some embodiments, the compounds disclosed herein bind to TEAD4 and prevent or inhibit the interaction of YAP and TEAD4.

In some embodiments, the compounds disclosed herein bind to TEAD1 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C359 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C53 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C327 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C405 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C359 and C327 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C359 and C53 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C53 and C327 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C359 and C405 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C53 and C405 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C327 and C405 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C359, C327, and C53 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C359, C327, and C405 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C359, C353, and C405 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C327, C53, and C405 and prevent or inhibit the interaction of YAP and TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1 at C359, C327, C53, and C405 and prevent or inhibit the interaction of YAP with TEAD1.

In some embodiments, the compounds disclosed herein bind to TEAD, prevent palmitoylation of TEAD, and prevent or inhibit the interaction of YAP and TEAD. In some embodiments, the compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1. In some embodiments, a compound disclosed herein binds to TEAD1 and prevents palmitoylation of TEAD1 at C359. In some embodiments, the compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1 at C53. In some embodiments, the compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1 at C327. In some embodiments, the compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1 at C405. In some embodiments, the compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1 at C359 and C327. In some embodiments, the compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1 at C359 and C53. In some embodiments, the compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1 at C53 and C327. In some embodiments, compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1 at C359 and C405. In some embodiments, the compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1 at C53 and C405. In some embodiments, compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1 at C327 and C405. In some embodiments, compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1 at C359, C327, and C53. In some embodiments, the compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1 at C359, C327, and C405. In some embodiments, the compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1 at C359, C353, and C405. In some embodiments, compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1 at C327, C53, and C405. In some embodiments, the compounds disclosed herein bind to TEAD1 and prevent palmitoylation of TEAD1 at C359, C327, C53, and C405.

In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1, and prevent or inhibit the interaction of YAP with TEAD1. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C359, and prevent or inhibit the interaction of TEAD1 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C53, and prevent or inhibit the interaction of TEAD1 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C327, and prevent or inhibit the interaction of TEAD1 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C405, and prevent or inhibit the interaction of TEAD1 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C359 and C327, and prevent or inhibit the interaction of TEAD1 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C359 and C53, and prevent or inhibit the interaction of TEAD1 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C53 and C327, and prevent or inhibit the interaction of TEAD1 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C359 and C405, and prevent or inhibit the interaction of TEAD1 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C53 and C405, and prevent or inhibit the interaction of TEAD1 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C327 and C405, and prevent or inhibit the interaction of TEAD1 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C359, C327, and C53, and prevent or inhibit the interaction of TEAD1 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C359, C327, and C405, and prevent or inhibit the interaction of TEAD1 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C359, C353, and C405, and prevent or inhibit the interaction of TEAD1 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C327, C53, and C405, and prevent or inhibit the interaction of TEAD1 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD1, prevent palmitoylation of TEAD1 at C359, C327, C53, and C405, and prevent or inhibit the interaction of TEAD1 with YAP.

In some embodiments, the compounds disclosed herein bind to TEAD2 at C380 and prevent or inhibit the interaction of YAP and TEAD2.

In some embodiments, the compounds disclosed herein bind to TEAD2 and prevent palmitoylation of TEAD2. In some embodiments, the compounds disclosed herein bind to TEAD2 and prevent palmitoylation of TEAD2 at C380.

In some embodiments, the compounds disclosed herein bind to TEAD2, prevent palmitoylation of TEAD2, and prevent or inhibit the interaction of YAP with TEAD2. In some embodiments, the compounds disclosed herein bind to TEAD2, prevent palmitoylation of TEAD2 at C380, and prevent or inhibit the interaction of TEAD2 with YAP.

In some embodiments, the compounds disclosed herein bind to TEAD3 at C371 and prevent or inhibit the interaction of YAP and TEAD3. In some embodiments, the compounds disclosed herein bind to TEAD3 at C368 and prevent or inhibit the interaction of YAP and TEAD3. In some embodiments, the compounds disclosed herein bind to TEAD3 at C371 and C368 and prevent or inhibit the interaction of YAP and TEAD3.

In some embodiments, the compounds disclosed herein bind to TEAD3 and prevent palmitoylation of TEAD3. In some embodiments, the compounds disclosed herein bind to TEAD3 and prevent palmitoylation of TEAD3 at C371. In some embodiments, the compounds disclosed herein bind to TEAD3 and prevent palmitoylation of TEAD3 at C368. In some embodiments, compounds disclosed herein bind to TEAD3 and prevent palmitoylation of TEAD3 at C368 and C371.

In some embodiments, the compounds disclosed herein bind to TEAD3, prevent palmitoylation of TEAD3, and prevent or inhibit the interaction of YAP and TEAD3. In some embodiments, the compounds disclosed herein bind to TEAD3, prevent palmitoylation of TEAD3 at C371, and prevent or inhibit the interaction of TEAD3 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD3, prevent palmitoylation of TEAD3 at C368, and prevent or inhibit the interaction of YAP with TEAD3. In some embodiments, the compounds disclosed herein bind to TEAD3, prevent palmitoylation of TEAD3 at C371 and C368, and prevent or inhibit the interaction of YAP with TEAD3.

In some embodiments, the compounds disclosed herein bind to TEAD4 at C367 and prevent or inhibit the interaction of YAP and TEAD4.

In some embodiments, compounds disclosed herein bind to TEAD4 and prevent palmitoylation of TEAD4. In some embodiments, the compounds disclosed herein bind to TEAD4 and prevent palmitoylation of TEAD4 at C367.

In some embodiments, the compounds disclosed herein bind to TEAD4, prevent palmitoylation of TEAD4, and prevent or inhibit the interaction of TEAD4 with YAP. In some embodiments, the compounds disclosed herein bind to TEAD4, prevent palmitoylation of TEAD4 at C367, and prevent or inhibit the interaction of TEAD4 with YAP.

YAP/TAZ Regulation Mediated by G Proteins/GPCRs In some embodiments, the Hippo pathway regulates G protein-coupled receptors (GPCRs) and members of the G protein (also known as guanine nucleotide binding proteins) family. Controlled by proteins. G proteins are molecular switches that transmit extracellular stimuli into cells via GPCRs. In some cases, there are two classes of G proteins: monomeric small GTPases and heterotrimeric G protein complexes. In some cases, the latter class of complexes includes alpha (G _α ), beta (G _β ), and gamma (G _γ ) subunits. In some cases, several classes of G _α subunits are present: G _q/11 α, G _12/13 α, G _i/o α (G-inhibitory, G-other), and G _s α (G-stimulatory). do.

In some cases, Giα (G-inhibitory), G _o α (G-other), G _q/ 11α, and G12/13α-coupled GPCRs activate YAP/TAZ and promote nuclear translocation. In other cases, G _s α (G-stimulated) coupled GPCRs suppress YAP/TAZ activity, resulting in YAP/TAZ degradation.

In some cases, G _i α (G-inhibitory), G _o α (G-other), G _q/11 α, and G _12/13 α-coupled GPCRs activate YAP/TAZ through suppression of Lats1/2 activity. . In contrast, G _s α, in some embodiments, induces Lats1/2 activity, thereby promoting YAP/TAZ degradation.

G _q family G _q α (also known as G _q/11 proteins) is responsible for the inositol triphosphate (IP ₃ ) signal transduction pathway and from intracellular stores through activation of phospholipase C (PLC). It is involved in the release of calcium (Ca ²⁺ ). The activated PLC hydrolyzes phosphatidylinositol 4,5-biphosphate (PIP ₂ ) to diacylglycerol (DAG) and IP ₃ . In some cases, _IP3 then diffuses through the cytoplasm to the ER or, in the case of muscle cells, into the sarcoplasmic reticulum (SR) and then to the inositol triphosphate receptor (InsP3R), a Ca2 ⁺ channel. Join. In some cases, binding causes the opening of Ca ²⁺ channels, which increases the release of Ca ²⁺ into the cytoplasm.

In some embodiments, GPCRs that interact with G _q α include 5-hydroxytryptamine receptors (5-HT receptors) types 5-HT ₂ and 5-HT ₃ ; α-1 adrenergic receptors ; vasopressin type 1 receptors 1A and 1B; angiotensin II receptor type 1; calcitonin receptor; histamine H1 receptor; metabotropic glutamate receptor group I; muscarinic receptors M ₁ , M ₃ , and M ₅ ; These include, but are not limited to, trace amine related receptor 1.

In some cases, there are several types of G _q α: G _q , G _q/11 , G _q/14 , and G _q/15 . _Gq protein is encoded by GNAQ. G _q/11 is coded by GNA11. G _q/14 is coded by GNA14. G _q/15 is coded by GNA15.

In some cases, mutations or modifications of the G _q α gene are associated with cancer. Indeed, mutations in G _q α have been shown to promote uveal melanoma (UM) tumorigenesis. In some cases, approximately 80% of UM cases have been detected to contain mutations in GNAQ and/or GNA11.

In some cases, mutations or modifications of the G _q α gene are associated with congenital diseases. In some cases, mutations in G _q α have been observed in congenital diseases such as port-wine nevi and/or Sturge-Weber syndrome. In some cases, approximately 92% of port wine nevus cases have mutations in GNAQ. In some cases, about 88% of people with Sturge-Weber syndrome have mutations in GNAQ.

G _12/13 family G _12/13 α regulates actin cytoskeletal remodeling in cells and controls cellular processes mediated by guanine nucleotide exchange factors (GEFs). GEFs are involved in the activation of small GTPases that function as molecular switches in various intracellular signal transduction pathways. Examples of small GTPases include the Ras-related GTPase superfamily (e.g., Rho family such as Cdc42), which are involved in cell differentiation, proliferation, cytoskeletal organization, vesicle trafficking, and nuclear transport. There is.

In some embodiments, GPCRs that interact with G _12/13 α include purinergic receptors (e.g., P2Y ₁ , P2Y ₂ , P2Y ₄ , P2Y ₆ ); muscarinic acetylcholine receptors M1 and M3; Receptors for thrombin [protease-related receptors (PAR)-1, PAR-2]; thromboxane (TXA2); sphingosine 1-phosphate (e.g., S1P ₂ , S1P ₃ , S1P ₄ , and S1P ₅ ); lysophosphatidic acid (e.g. LPA ₁ , LPA ₂ , LPA ₃ ); angiotensin II (AT1); serotonin (5-HT _2c and 5-HT ₄ ); somatostatin (sst ₅ ); endothelin (ET _A and ET _B ); cholecystokinin (CCK ₁ ); V _1a vasopressin receptor; D ₅ dopamine receptor; fMLP formyl peptide receptor; GAL ₂ galanin receptor; EP ₃ prostanoid receptor; A ₁ adenosine receptor; α ₁ adrenergic receptor; _BB2 bombesin receptor; _B2 bradykinin receptor; calcium-sensitized receptor; KSHV-ORF74 chemokine receptor; _NK1 tachykinin receptor; and thyroid stimulating hormone (TSH) receptor. .

In some cases, G _12/13 α is further subdivided into G ₁₂ and G ₁₃ types, encoded by GNA12 and GNA13, respectively.

G _i/o family G _i/o α (G-inhibitory, G-other) (also known as G _i /G _o or G _i proteins) acts through inhibition of adenylate cyclase activity, which converts ATP to cAMP. , suppresses the production of 3',5'-cyclic AMP (cAMP) from adenosine triphosphate (ATP).

In some embodiments, the GPCRs that interact with G _iα include 5-hydroxytryptamine receptor (5- _HT receptor) types 5-HT ₁ and 5-HT _{5 ;} acetylcholine receptors; adenosine receptors such as A ₁ and A ₃ ; adrenergic receptors such as α _2A , α _2B , and α _2c ; apelin receptors; calcium-sensitized receptors; cannabinoid receptors CB1 and CB2; chemokines CXCR4 receptor; dopamine D ₂ , D ₃ , and D ₄ ; GABA _B receptor; metabotropic glutamate receptor 2 (mGluR2), metabotropic glutamate receptor 3 (mGluR3), metabotropic glutamate receptor 4 ( glutamate receptors such as metabotropic glutamate receptor 6 (mGluR6), metabotropic glutamate receptor 7 (mGluR7), and metabotropic glutamate receptor 8 (mGluR8); _H3 and _H4 receptors, etc. histamine receptors; melatonin receptors such as melatonin receptor type 1 (MT1); melatonin receptor type 2 (MT2), and melatonin receptor type 3 (MT3); niacin receptors such as NIACR1 and NIACR2; δ, κ , μ, and nociceptin receptors; prostaglandin E receptor 1 (EP ₁ ), prostaglandin E receptor 3 (EP ₃ ), prostaglandin F receptor (FP), and thromboxane somatostatin receptors sst ₁ , sst ₂ , sst ₃ , sst ₄ , and sst ₅ ; and trace amine-related receptor 8.

In some cases, there are several types of G _i α; G _i α1, G _i α2, G _i α3, G _i α4, G _o α, G _t , G _gust , and G _z . G _i α1 is encoded by GNAI1. G _i α2 is encoded by GNAI2. G _i α3 is encoded by GNAI3. The α _o subunit, G _o α, is encoded by GNAO1. G _t is encoded by GNAT1 and GNAT2. G _gust is coded by GNAT3. G _z is coded by GNAZ.

G _s family G _s α (also known as G stimulatory, G _s alpha subunit, or G _s protein) converts adenosine triphosphate (ATP) into 3'5'-cyclic AMP (cAMP) and pyroproteins. Activates the cAMP-dependent pathway through activation of adenylate cyclase, which converts to phosphate. In some embodiments, GPCRs that interact with G _s α include 5-hydroxytryptamine receptors (5-HT receptors) types 5-HT ₄ , 5-HT ₆ , and 5-HT ₇ ; Stimulating hormone receptor (ACTH receptor) (also known as melanocortin receptor 2 or MC2R); adenosine receptor types A _2a and A _2b ; arginine vasopressin receptor 2 (AVPR2); β-adrenergic receptor β ₁ , β ₂ , and β ₃ ; calcitonin receptor; calcitonin gene-related peptide receptor; dopamine receptor D1-like family receptors such as corticotropin-releasing hormone receptor, D ₁ and D ₅ ; follicle-stimulating hormone receptor gastric inhibitory polypeptide receptors; glucagon receptors; histamine _H2 receptors; luteinizing hormone/choriogonadotropin receptors; melanocortin receptors such as MC1R, MC2R, MC3R, MC4R, and MC5R; These include, but are not limited to, parathyroid hormone receptor 1; prostaglandin receptor types D ₂ and I ₂ ; secretin receptor; thyrotropin receptor; trace amine-related receptor 1; and cubic jellyfish opsin. .

In some cases, there are two types of G _s α: G _s and G _olf . G _s is coded by GNAS. _Golf is coded by GNAL.

Additional Regulators of the Hippo Signaling Network In some embodiments, the additional regulator of the Hippo signaling pathway is the Crumbs (Crb) complex. The Crumbs complex is a key regulator of cell polarity and cell shape. In some cases, the Crumbs complex includes transmembrane CRB proteins that assemble multi-protein complexes that function in cell polarity. In some cases, the CRB complex recruits members of the angiomotin (AMOT) family of adapter proteins, which interact with Hippo pathway components. Studies have shown that in some cases, AMOT binds directly to YAP, promoting YAP phosphorylation and inhibiting its nuclear localization.

In some cases, additional regulators of the Hippo signaling pathway include regulators of the MST kinase family. MST kinase monitors the integrity of the actin cytoskeleton. In some cases, the regulatory factors include TAO kinase and cell polarity kinase PAR-1.

In some cases, additional regulators of the Hippo signaling pathway include molecules of adherens junctions. In some cases, E-cadherin (E-cad) suppresses the nuclear localization and activity of YAP by regulating MST activity. In some embodiments, the E-cad-related protein a-catenin regulates YAP by sequestering the YAP/14-3-3 complex within the cytoplasm. In other cases, Ajuba protein family members interact with Lats1/2 kinase activity, thereby preventing YAP/TAZ deactivation.

In some embodiments, additional proteins that interact either directly or indirectly with YAP/TAZ include Merlin, protocadherin Fat1, mask 1/2, HIPK2, PTPN14, RASSF, PP2A, salt Examples include, but are not limited to, inducible kinase (SIK), Scribble (SCRIB), Scribble-related protein Discs large (Dlg), KIBRA, PTPN14, NPHP3, LKB1, Ajuba, and ZO1/2.

In some embodiments, the compounds described herein are inhibitors of transcriptional coactivators of PDZ-binding motif/Yes-associated protein transcriptional coactivators (TAZ/YAP). In some embodiments, the compounds described herein increase the phosphorylation of transcriptional coactivator of PDZ-binding motif/Yes-associated protein transcriptional coactivator (TAZ/YAP), or Reduces dephosphorylation of transcriptional coactivator/Yes-related protein coactivator (TAZ/YAP). In some embodiments, the compound increases ubiquitination of a transcriptional coactivator by PDZ-binding motif/Yes-associated protein coactivator (TAZ/YAP) or Reduces deubiquitination of transcriptional coactivator by activator (TAZ/YAP).

In some embodiments, a compound disclosed herein is an inhibitor of one or more of the proteins involved in or associated with the Hippo pathway. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of a G protein and/or its associated GPCR. In some embodiments, the Hippo pathway inhibitor is a G protein inhibitor. In some embodiments, inhibitors of the Hippo pathway are G _q α family proteins, such as G _q , G _q/11 , G _{q/14 ,} and G _{q/ 15 ; /13} α family protein; or is an inhibitor of G _i α family proteins of G _i α1, G _i α2, G _i α3, G _i α4, G _o α, G _t , G _gust and G _z . In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of G _q . In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of G _q/11 . In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of G _q/14 . In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of G _q/15 . In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of _G12 . In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of _G13 . In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of G _i α1. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of G _i α2. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of G _i α3. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of G _i α4. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of G _o α. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of _Gt . In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of G _gust . In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of _Gz .

In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of the Hippo pathway core protein. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of Sav1. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of Mob. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of YAP. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of TAZ. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of TEAD.

In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of a protein associated with the ubiquitination and proteasomal degradation pathway. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of a proteasomal degradation pathway protein (eg, 26S proteasome).

In some embodiments, the inhibitor of the Hippo pathway is a protein inhibitor of the Ras superfamily of proteins. In some embodiments, the inhibitor of the Hippo pathway is a protein inhibitor of the Rho family of proteins. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of Cdc42.

Cdc42 is a member of the Ras superfamily of small GTPases. Specifically, Cdc42 belongs to the Rho family of GTPases, members of which are involved in diverse and important cellular processes such as gene transcription, cell-cell adhesion, and cell cycle progression. Cdc42 is involved in cell proliferation and polarity, and in some cases Cdc42 is activated by guanine nucleotide exchange factors (GEFs). In some cases, the inhibitor of Cdc42 is a compound disclosed herein.

In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of a deubiquitinating enzyme. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of cysteine proteases or metalloproteases. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of a ubiquitin-specific protease. USP47 is a member of the ubiquitin-specific protease (USP/UBP) family of cysteine proteases. In some embodiments, compounds disclosed herein are inhibitors of USP47.

In some embodiments, the invention provides the use of a compound, or a pharmaceutical salt or composition thereof, to treat one or more disorders, diseases, and/or medical conditions, including Or pathological conditions include, but are not limited to, cell proliferative disorders.

The activity of compounds utilized in the invention as inhibitors of TEADs (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4), or variants or mutants thereof, can be determined in vitro, in vivo, or in cell lines. can be assayed. In vitro assays include assays that measure inhibition of TEADs (eg, TEAD1, TEAD2, TEAD3, and/or TEAD4), or variants or mutants thereof. An alternative in vitro assay quantifies the ability of an inhibitor to bind to a TEAD (eg, TEAD1, TEAD2, TEAD3, and/or TEAD4) or a variant or variant thereof. Detailed conditions for assaying compounds utilized in the present invention as inhibitors of TEADs (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4), or variants or mutants thereof, are described below. Let's explain with an example. See, eg, Examples 2 and 5.

As used herein, the terms "treatment", "treating", and "treating" refer to reversing or ameliorating the disease or disorder described herein, or one or more symptoms thereof. This refers to delaying the onset or inhibiting the progression of the disease. In some embodiments, treatment may be performed after one or more symptoms have developed. In other embodiments, treatment may be performed in the absence of symptoms. For example, treatment may be administered to susceptible individuals prior to the onset of symptoms (eg, determined in terms of history of symptoms and/or in terms of genetic or other susceptibility factors). Treatment may also be continued after symptoms have remitted, eg, to prevent or delay their recurrence.

Provided compounds are inhibitors of TEADs (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4) and, therefore, are associated with the activity of TEADs (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4). is useful for treating one or more disorders in which Accordingly, in certain aspects and embodiments, the invention provides a step of administering a therapeutically effective compound of the invention, or a pharmaceutically acceptable composition thereof, to a patient in need of treatment for a TEAD-mediated disorder. Provided are methods of treating TEAD-mediated disorders including:

As used herein, the term "TEAD-mediated" disorder, disease, and/or condition refers to a TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4) or any disease or other deleterious condition in which these variants or variants are known to play a role. Accordingly, another aspect or embodiment of the invention is to provide a TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4), or one of the variants or variants thereof known to play a role. The invention relates to treating the above-mentioned diseases or reducing the severity of the diseases.

As used herein, the term "therapeutically effective amount" refers to reducing or attenuating the biological activity of a variant, conferring a therapeutic effect in the treatment of a medical condition, or in a biological sample or patient. an amount of a TEAD inhibitor, or a pharmaceutically acceptable salt thereof, effective to delay or minimize one or more symptoms associated with a medical condition; TEAD4) or a variant thereof. In some embodiments, a "therapeutically effective amount" refers to the binding or signaling activity of a TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4), or a variant or variant thereof; Alternatively, it refers to an amount of a TEAD inhibitor or pharmaceutically acceptable salt thereof that measurably reduces any TEAD-mediated activity. The term "therapeutically effective amount" includes, in some embodiments, the treatment of another therapeutic agent that improves the overall treatment regimen, reduces or avoids the symptoms, signs, or causes of a medical condition, and/or Effect-enhancing amounts can be included. In certain embodiments, a therapeutically effective amount is an amount sufficient to inhibit a TEAD transcription factor. In certain embodiments, a therapeutically effective amount is an amount sufficient to treat a proliferative disease.

In some aspects and embodiments, a therapeutically effective compound of the invention, or a pharmaceutically acceptable composition thereof, is administered to a patient in need thereof. increasing the expression of a TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4) and/or increasing the expression of a TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4) methods of treating, reducing the severity of, or reducing the severity of, or one or more symptoms of, a disease or disorder characterized by or associated with increased activity. Methods of delaying or inhibiting their progression are provided herein. In some aspects and embodiments, a therapeutically effective compound of the invention, or a pharmaceutically acceptable composition thereof, is administered to a patient in need thereof. any disease or disorder in which inhibition or antagonism of TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4) activity is beneficial, or one or more of Provided herein are methods of treating the symptoms, reducing their severity, delaying their onset, or inhibiting their progression. In some aspects and embodiments, a therapeutically effective compound of the invention, or a pharmaceutically acceptable composition thereof, is administered to a patient in need thereof. A method of treating a disease or disorder, or one or more symptoms thereof, in which inhibition or antagonism of the Hippo pathway would be beneficial, a method of reducing the severity thereof, comprising administering a composition thereof that is acceptable to the public. Provided herein are methods for delaying the onset of or inhibiting the progression of.

In some aspects and embodiments, the invention provides a method for administering a TEAD inhibitor compound described herein to a patient in need thereof, or a pharmaceutical salt or composition thereof. , or pharmaceutical salts or compositions thereof, including but not limited to cell proliferative disorders. . In some embodiments, the cell proliferative disorder is cancer. In some embodiments, the cancer has increased expression of TEADs (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4); ) characterized by increased activity.

As used herein, the terms "increased," "elevated," or "enhanced" are used interchangeably and include biological functions and/or Includes any measurable increase in biological activity and/or concentration. For example, the increase is at least about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, compared to a control or baseline amount of function, activity, or concentration; about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97 %, approximately 98%, approximately 99%, approximately 100%, approximately 2 times, approximately 3 times, approximately 4 times, approximately 5 times, approximately 6 times, approximately 7 times, approximately 8 times, approximately 9 times, approximately 10 times, It may be about 20 times, about 25 times, about 50 times, about 100 times, or more.

As used herein, the term "increased expression" and/or "increased activity" of a substance, e.g., a TEAD, in a sample or cancer or patient is measured by techniques known in the art. about 5%, about 10% compared to the amount of a substance, such as a TEAD, in a control sample(s), such as an individual or population that does not suffer from a disease or disorder (e.g., cancer), or an internal control. , about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, approximately 80%, approximately 85%, approximately 90%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, approximately 99%, approximately 100%, approximately 2 times, approximately 3 times, approximately 4 times , about 5 times, about 6 times, about 7 times, about 8 times, about 9 times, about 10 times, about 20 times, about 25 times, about 50 times, about 100 times, or more, such as TEAD means an increase in the amount of The expression and/or activity of TEAD compared to the mean (average) or median amount of TEAD in a control group of samples, or a baseline group of samples, or a retrospective study of patient samples. A subject is said to have "increased expression" or "increased activity" of a TEAD if it increases by 1 standard deviation, 2 standard deviations, 3 standard deviations, 4 standard deviations, 5 standard deviations, or more. It can also be measured. As practiced in the art, such control or baseline expression levels may be predetermined or measured prior to measurement in the sample or cancer or subject, or may be obtained from a database of such control samples. Obtainable.

As used herein, "proliferative disease" means a disease caused by abnormal growth or expansion of cells (Walker, Cambridge Dictionary of Biology, Cambridge University Press: Cambridge, UK, 1990). ). Proliferative diseases include 1) pathological proliferation of normal quiescent cells; 2) pathological migration of cells from their normal location (e.g. metastasis of neoplastic cells); 3) matrix metalloproteinases (e.g. collagenase, or 4) may be associated with pathological angiogenesis, such as in proliferative retinopathy and tumor metastasis. Exemplary proliferative diseases include cancer (ie, "malignant tumors"), benign tumors, angiogenesis, inflammatory diseases, and autoimmune diseases.

Cancer Cancers or proliferative disorders or tumors treated with the compounds and methods and uses described herein include hematological cancers, lymphomas, myeloma, leukemias, neurological cancers, skin cancers, breast cancers, prostate cancers. cancer, including, but not limited to, colorectal cancer, lung cancer, head and neck cancer, gastrointestinal cancer, liver cancer, pancreatic cancer, genitourinary cancer, osteosarcoma, renal cancer, and vascular cancer.

In some embodiments of the methods and uses described herein, the cancer is mediated by transcriptional coactivator activation by PDZ-binding motif/Yes-associated protein transcriptional coactivator (TAZ/YAP). be done. In some embodiments of the methods and uses described herein, the cancer is mediated by modulating the interaction of YAP/TAZ with TEADs (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4). be done. In some embodiments of the methods and uses described herein, the cancer is characterized by increased TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4) expression; , TEAD2, TEAD3, and/or TEAD4) activity. In some embodiments of the methods and uses described herein, the cancer is a cancer in which YAP is localized to the nucleus of cancer cells.

In some embodiments of the methods and uses described herein, the cancer is characterized by or associated with genetic alterations in one or more Hippo pathway genes. As used herein, the term "genetic alteration in one or more Hippo pathway genes" refers to a change in a sample, such as a tumor sample, that has a detectable amount of genetic alteration in one or more Hippo pathway genes. Mean percentage cells. As used herein, genetic changes in genes, such as Hippo pathway genes, include, for example, loss-of-function mutations within the gene (including, for example, frameshifts, nonsense mutations, and splicing mutations), changes in gene copy number. (eg, a copy number gain, amplification, copy number loss, or deletion) or a fusion of the gene with another gene, such as a TAZ-CAMTA1 fusion or a YAP1-TFE3 fusion. In some embodiments, a genetic alteration in a Hippo pathway gene is about 5%, about 10%, about 15%, about 20%, about 25%, about 30% of cells, such as tumor cells, in a sample. %, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% of at least about 3 copies of the genetically engineered Hippo pathway gene; at least about 4 copies of the genetically engineered Hippo pathway gene; a genetically engineered Hippo pathway gene, at least about 5 copies of a genetically engineered Hippo pathway gene, at least about 6 copies of a genetically engineered Hippo pathway gene, at least about 7 copies of a genetically engineered Hippo pathway gene, at least about 8 copies of a genetically engineered Hippo pathway gene; , a genetically engineered Hippo pathway gene, at least about 9 copies of a genetically engineered Hippo pathway gene, at least about 10 copies of a genetically engineered Hippo pathway gene, at least about 11 copies of a genetically engineered Hippo pathway gene. , at least about 12 copies of a genetically engineered Hippo pathway gene, at least about 9 copies of a genetically engineered Hippo pathway gene, at least about 10 copies of a genetically engineered Hippo pathway gene, at least about 11 copies of a gene. a recombinant Hippo pathway gene, at least about 12 copies of a recombinant Hippo pathway gene, at least about 13 copies of a recombinant Hippo pathway gene, at least about 14 copies of a recombinant Hippo pathway gene, at least Means having about 15 copies of the genetically engineered Hippo pathway gene, at least about 20 copies of the genetically engineered Hippo pathway gene, or more. In some embodiments, a genetic alteration in a Hippo pathway gene means that about 10% of tumor cells in the sample have at least about 15 copies of a genetically modified Hippo pathway gene. In some embodiments, a genetic alteration in a Hippo pathway gene means that about 40% of tumor cells in the sample have at least about 4 copies of a genetically modified Hippo pathway gene. In some embodiments, a genetic alteration in a Hippo pathway gene means that about 10% of tumor cells in the sample have at least about 4 copies of a genetically modified Hippo pathway gene. In some embodiments, the Hippo pathway gene is NF2. In some embodiments, the genetic alteration in one or more Hippo pathway genes is NF2 deficiency. In some embodiments, NF2 deficiency refers to functionally mutated NF2 loss. In some embodiments, NF2 deficiency refers to loss or deletion of NF2 copies. In some embodiments, NF2 deficient means that NF2 mRNA expression is absent or very low. In some embodiments, the Hippo pathway gene is YAP1. In some embodiments, the genetic change in one or more Hippo pathway genes is YAP1 amplification. In some embodiments, the genetic change in one or more Hippo pathway genes is a YAP1 fusion, such as a YAP1-TFE3 fusion. In some embodiments, the Hippo pathway gene is TAZ. In some embodiments, the genetic change in one or more Hippo pathway genes is a TAZ amplification. In some embodiments, the genetic change in one or more Hippo pathway genes is a TAZ fusion, such as a TAZ-CAMTA1 fusion. In some embodiments, the Hippo pathway gene is LATS 1/2. In some embodiments, the genetic alteration in one or more Hippo pathway genes is a LATS 1/2 copy number loss or deletion. In some embodiments, the Hippo pathway gene is MST1/2. In some embodiments, the Hippo pathway gene is BAP1.

In some embodiments, the cancer is characterized by a mutant Gα-protein. In some embodiments, the variant Gα-protein is selected from G12, G13, Gq, G11, Gi, Go, and Gs. In some embodiments, the variant Gα-protein is G12. In some embodiments, the Gα-protein is G13. In some embodiments, the Gα-protein is Gq. In some embodiments, the Gα-protein is G11. In some embodiments, the Gα-protein is Gi. In some embodiments, the Gα-protein is Go. In some embodiments, the Gα-protein is Gs.

In some embodiments of the methods and uses described herein, cancer is treated by inhibiting or reducing or reducing or stopping further growth or spread of the cancer or tumor. . In some embodiments of the methods and uses described herein, the cancer is at least 5%, at least 10%, at least 25%, at least 50% smaller than the size of the cancer or tumor before treatment. inhibiting or reducing the size (e.g., volume or mass) of a cancer or tumor by at least 75%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%. Treated by In some embodiments of the methods and uses described herein, the cancer is at least 5%, at least 10%, at least 25%, at least 50% as compared to the amount of cancer or tumor before treatment. , at least 75%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, by reducing the amount of cancer or tumor in the patient.

In some embodiments, the patient treated using the methods or uses described herein is at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, after initiation of treatment. at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 18 months, at least about 2 years, Demonstrates a progression free survival of at least about 3 years, at least about 4 years, or at least about 5 years. In some embodiments, the patient treated using the methods or uses described herein is at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, after initiation of treatment. at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 14 months, at least about 16 months, Demonstrates an overall survival time of at least about 18 months, at least about 20 months, at least about 22 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years.

In some embodiments, the patient treated using the methods or uses described herein has at least about 15%, at least about 20%, at least about 25%, at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100 %, objective overall efficiency (ORR).

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 It is 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 cancer. Hemangioendothelioma (EHE). In some embodiments, the cancer is mesothelioma, such as malignant mesothelioma. In some embodiments, the cancer is EHE.

In some embodiments, the cancer includes, but is not limited to, leukemia (e.g., acute leukemia, acute lymphocytic leukemia, acute myeloid leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myeloid leukemia). monocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia), polycythemia vera, lymphoma (e.g., Hodgkin's disease or non-Hodgkin's disease), Waldenstrees muma macroglobulinemia, multiple myeloma, heavy chain disease, and solid tumors such as sarcomas and cancers (e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, Internal sarcoma, lymphangiosarcoma, intralymphatic sarcoma, synoviomas, mesothelioma, Ewing tumor, leiomyosarcoma, 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, bronchial carcinoma, renal cell carcinoma, liver cancer, cholangiocarcinoma, choriocarcinoma, seminoma, embryonal carcinoma , Wilms tumor, cervical cancer, uterine cancer, testicular cancer, lung cancer, small cell lung cancer, bladder cancer, epithelial cancer, glioma, astrocytoma, glioblastoma multiforme (GBM, also known as glioblastoma). medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma, melanoma, nerve blastoma, and retinoblastoma).

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

In some embodiments, the cancer is an acoustic neuroma, an astroglioma (e.g., grade I - pilocytic astrocytoma, grade II - low-grade astrocytoma, grade III - undifferentiated astrocytoma or grade IV-glioblastoma (GBM)), chordoma, CNS lymphoma, craniopharyngioma, brainstem glioma, ependymoma, mixed glioma, optic glioma, subependymoma, medullary blastoma, meningioma, metastatic brain tumor, oligodendroglioma, pituitary tumor, undifferentiated neuroectodermal (PNET) tumor, or schwannoma. In some embodiments, the cancer is a brainstem glioma, craniopharyngioma, ventricular ependymoma, juvenile pilocytic astrocytoma (JPA), medulloblastoma, optic glioma, pineal region tumor. , primitive neuroectodermal tumors (PNETs), or rhabdoid tumors, which are types more commonly found in children than adults. Children In some embodiments, the patient is an adult human. In some embodiments, the patient is a child or pediatric patient.

Cancers, in another embodiment, include, but are not limited to, mesothelioma, hepatobiliary (liver and bile ducts), bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular melanoma, and ovarian cancer. , colon cancer, rectal cancer, cancer of the anal region, stomach cancer, gastrointestinal (stomach, colorectal, and duodenum), uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, Esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, 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's lymphoma, spinal axis tumor, brainstem glioma, pituitary adenoma, adrenocortical carcinoma, gallbladder cancer, multiple myeloma, bile duct carcinoma, Includes fibrosarcoma, neuroblastoma, retinoblastoma, or a combination of one or more of the aforementioned cancers.

In some embodiments, the cancer is hepatocellular carcinoma, ovarian cancer, epithelial ovarian cancer, or fallopian tube cancer; papillary serous cystadenocarcinoma or uterine papillary serous carcinoma (UPSC); prostate cancer; testicular cancer Cancer; gallbladder cancer; bile duct hepatocarcinoma; soft tissue and osteosynovial sarcoma; rhabdomyosarcoma; osteosarcoma; chondrosarcoma; Ewing's sarcoma; undifferentiated thyroid carcinoma; adrenocortical adenoma; pancreatic cancer; pancreatic ductal or pancreatic adenocarcinoma; / gastric (GIST) cancer; lymphoma; squamous cell carcinoma of the head and neck (SCCHN); salivary gland cancer; glioma, or brain cancer; neurofibromatosis-1-related malignant peripheral nerve sheath tumor (MPNST); Waldenström selected from macroglobulinemia; or medulloblastoma.

In some embodiments, the cancer is hepatocellular carcinoma (HCC), hepatoblastoma, colon cancer, rectal cancer, ovarian cancer, epithelial ovarian cancer, fallopian tube cancer, papillary serous cystadenocarcinoma, uterine papillary cancer. serous carcinoma (UPSC), bile duct hepatocarcinoma, soft tissue and osteosynovial sarcoma, rhabdomyosarcoma, osteosarcoma, undifferentiated thyroid carcinoma, adrenocortical adenoma, pancreatic cancer, pancreatic ductal carcinoma, pancreatic adenocarcinoma, glioma, selected from neurofibromatosis-1 associated malignant peripheral nerve sheath tumor (MPNST), Waldenström macroglobulinemia, or medulloblastoma.

In some embodiments, the cancer is a solid tumor, such as a sarcoma, carcinoma, or lymphoma. Tumors usually include abnormal masses of tissue that typically do not contain cysts or areas of fluid. In some embodiments, the cancer is renal cell carcinoma, or kidney cancer; hepatocellular carcinoma (HCC) or hepatoblastoma, or liver cancer; melanoma; breast cancer; colorectal carcinoma, or colorectal cancer; colon cancer; Cancer; anal cancer; lung cancer, such as non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC); ovarian cancer, epithelial ovarian cancer, ovarian carcinoma, or fallopian tube cancer; papillary serous cystadenocarcinoma or uterine papillary cancer prostatic serous carcinoma (UPSC); prostate cancer; testicular cancer; gallbladder cancer; bile duct hepatocarcinoma; soft tissue and bone synovial sarcoma; rhabdomyosarcoma; osteosarcoma; chondrosarcoma; Ewing's sarcoma; undifferentiated thyroid cancer; adrenocortical carcinoma ; pancreatic cancer; pancreatic ductal or pancreatic adenocarcinoma; gastrointestinal/stomach (GIST) cancer; lymphoma; squamous cell carcinoma of the head and neck (SCCHN); salivary gland cancer; glioma, or brain cancer; neurofibromatosis-1-related malignancies selected from peripheral nerve sheath tumor (MPNST); Waldenström macroglobulinemia; or medulloblastoma. stomach

In some embodiments, the cancer is renal cell carcinoma, hepatocellular carcinoma (HCC), hepatoblastoma, colorectal carcinoma, colorectal cancer, colon cancer, rectal cancer, anal cancer, ovarian cancer, epithelial ovarian cancer, ovarian cancer. Cancer, fallopian tube cancer, papillary serous cystadenocarcinoma, uterine papillary serous cystadenocarcinoma (UPSC), bile duct hepatocarcinoma, soft tissue and bone synovial sarcoma, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, undifferentiated thyroid cancer , adrenocortical carcinoma, pancreatic cancer, pancreatic ductal carcinoma, pancreatic adenocarcinoma, glioma, brain cancer, neurofibromatosis-1-related malignant peripheral nerve sheath tumor (MPNST), Waldenström macroglobulinemia, or medulla bud Selected from tumors.

In some embodiments, the cancer is hepatocellular carcinoma (HCC), hepatoblastoma, colon cancer, rectal cancer, ovarian cancer, epithelial ovarian cancer, ovarian cancer, fallopian tube cancer, papillary serous cystadenocarcinoma , uterine papillary serous carcinoma (UPSC), bile duct hepatocarcinoma, soft tissue and bone synovial sarcoma, rhabdomyosarcoma, osteosarcoma, undifferentiated thyroid carcinoma, adrenocortical carcinoma, pancreatic cancer, pancreatic ductal carcinoma, pancreatic adenocarcinoma, nerve selected from glioma, neurofibromatosis-1 associated malignant peripheral nerve sheath tumor (MPNST), Waldenström macroglobulinemia, or medulloblastoma.

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 ovarian carcinoma. 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 bile duct hepatocarcinoma. In some embodiments, the cancer is soft tissue and bone synovial sarcoma. In some embodiments, the cancer is rhabdomyosarcoma. In some embodiments, the cancer is osteosarcoma. In some embodiments, the cancer is anaplastic thyroid cancer. In some embodiments, the cancer is adrenocortical carcinoma. In some embodiments, the cancer is pancreatic cancer or pancreatic ductal cancer. In some embodiments, the cancer is pancreatic adenocarcinoma. In some embodiments, the cancer is a glioma. In some embodiments, the cancer is a malignant peripheral nerve sheath tumor (MPNST). In some embodiments, the cancer is neurofibromatosis-1 associated MPNST. In some embodiments, the cancer is Waldenström's macroglobulinemia. In some embodiments, the cancer is medulloblastoma.

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

In some embodiments, the cancer is melanoma cancer. 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).

Compounds and compositions may be administered according to the methods of the invention in any amount and using any route of administration effective to treat or reduce the severity of cancer or tumors. The exact amount required will vary from subject to subject depending on the species, age, and general condition of the subject, the severity of the disease or condition, the particular drug, its mode of administration, and the like. The compounds of the invention are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The expression "dosage unit form" as used herein refers to physically discrete units of the drug appropriate for the patient being treated. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The particular effective dose level for any particular patient or organism will depend on the disorder being treated and the severity of the disorder; the activity of the particular compound employed; the particular composition employed; the age, weight, and weight of the patient; general health, sex and diet; time of administration, route of administration, and rate of excretion of the particular compound used; duration of treatment; drugs used in combination or concomitantly with the particular compound used; Depends on a variety of factors, including similar factors well known in the field. The term "patient" or "subject" as used herein means an animal, preferably a mammal, and most preferably a human.

The pharmaceutically acceptable compositions of the invention may be administered orally, rectally, parenterally, intravesically, intravaginally, intraperitoneally, or topically, depending on the severity of the disease or disorder being treated. It can be administered to humans and other animals, such as intrabuccally (by powder, ointment, or drops), orally or as a nasal spray. In certain embodiments, the compounds of the invention are administered at a dosage of about 0.01 mg/kg to about 50 mg/kg of subject body weight and preferably about 1 mg/kg to about 25 mg of subject body weight per day to achieve the desired therapeutic effect. It may be administered orally or parenterally once or more per day at a dosage level of /kg subject body weight.

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

Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions, may be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution, suspension, or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. could be. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S. S. P. , and isotonic sodium chloride solutions. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or diglycerides. Additionally, fatty acids such as oleic acid can be used in the preparation of injectables.

Injectable preparations contain sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other sterile injectable media before use, for example, by filtration through a bacteria-retaining filter. By incorporating, it can be sterilized.

In order to prolong the effectiveness of the compounds of the invention, it is often desirable to delay the absorption of the compounds from subcutaneous or intramuscular injection. This can be achieved by the use of liquid suspensions of crystalline or amorphous materials that have poor water solubility. The rate of absorption of a compound therefore depends on its rate of dissolution, which in turn may depend on crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of compound to polymer and the nature of the particular polymer used, the rate of compound release can be adjusted. Other examples of biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.

Compositions for rectal or intravaginal administration preferably include a compound of the invention in a suitable non-irritating excipient or carrier, such as a solid at ambient temperature but liquid at body temperature, which may be administered rectally or intravaginally. Suppositories that can be prepared by mixing with cocoa butter, polyethylene glycol, or suppository wax, etc., dissolve in the vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is present in at least one inert pharmaceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate, and/or a) starch, lactose, sucrose. , fillers or extenders such as glucose, mannitol, and silicic acid; b) binders such as carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and acacia; c) wetting agents such as glycerol; d) disintegrants such as agar, calcium carbonate, potato starch or tapioca starch; e) solution retarders such as paraffin; f) absorption enhancers such as quaternary ammonium compounds; g) e.g. cetyl alcohol and glycerol monostearate. Wetting agents, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. For capsules, tablets, and pills, the dosage form may also include a buffer.

Solid compositions of a similar type may also be employed as fillers in soft- and hard-filled gelatin capsules using excipients such as lactose or milk sugar, and high molecular weight polyethylene glycols, and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical arts. These may optionally contain opacifying agents, and these may be of a composition that allows the active ingredient(s) to be released only, or preferably in certain parts of the intestinal tract, optionally with a delay. Can be done. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft- and hard-filled gelatin capsules using lactose or milk sugar as excipients, as well as high molecular weight polyethylene glycols and the like.

The active compounds may also be in micro-encapsulated form with one or more excipients mentioned above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, controlled release coatings, and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms, the active compound may be mixed with at least one inert diluent such as sucrose, lactose, or starch. According to conventional practice, such dosage forms may also contain additional substances other than inert diluents, such as tabletting lubricants and other tabletting aids, such as magnesium stearate and microcrystalline cellulose. For capsules, tablets, and pills, the dosage form may also include a buffering agent. They may optionally contain opacifying agents or may be of a composition that preferably releases the active ingredient(s) only in certain parts of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

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

Coadministration with one or more other therapeutic agent(s) Coadministration with one or more other therapeutic agent(s) Depending on the particular medical condition or disease being treated, the treatment of that condition Additional therapeutic agents normally administered for the purpose of treatment 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." .

In some embodiments, the invention comprises administering to a patient in need of treatment for a disclosed disease or condition an effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof. , co-administering an effective amount of one or more additional therapeutic agents such as those described herein, either simultaneously or sequentially. In some embodiments, the method includes co-administering one additional therapeutic agent. In some embodiments, the method includes co-administering two additional therapeutic agents. In some embodiments, the combination of a disclosed compound and additional therapeutic agent(s) acts synergistically.

The compounds of the invention can also be used in combination with known therapeutic processes, such as the administration of hormones or radiation. In certain embodiments, the provided compounds are used as radiosensitizers, particularly for treating tumors that exhibit insufficient sensitivity to radiotherapy.

The compounds of the invention can be administered alone or in combination with one or more other therapeutic compounds, and possible combination therapy includes administration of the compounds of the invention staggered or independently of each other. The combination may take the form of a fixed combination or administration with one or more other therapeutic compounds, or a fixed combination and one or more other therapeutic compounds. The compounds of the invention may be administered separately or in addition for tumor therapy, particularly in combination with chemotherapy, radiotherapy, immunotherapy, phototherapy, surgical intervention, or combinations thereof. be able to. As mentioned above, long-term therapy is equally possible, as is adjuvant therapy in the context of other treatment methods. Other possible treatments are treatments that maintain the patient's condition after tumor regression, or even, for example, chemopreventive therapy in at-risk patients.

One or more other therapeutic agent(s) can be administered separately from a compound or composition of the invention as part of a multiple dosing regimen. Alternatively, one or more other therapeutic agent(s) can be mixed with a compound of the invention in a single composition, which can be part of a single dosage form. When administered as a multiple dosing regimen, the one or more other therapeutic agent(s) and the compound or composition of the invention may be administered simultaneously, sequentially, or within a certain amount of time of each other, e.g. 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 from each other can be administered within In some embodiments, one or more other therapeutic agents (therapeutic agents) and a compound or composition of the invention are administered as a multiple dosage regimen within more than 24 hours apart.

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

A compound of the invention (in a composition comprising the additional therapeutic agent(s) described above) and one or more other therapeutic agent(s), which can be combined with carrier materials to produce a single dosage form. The amount will vary depending on the host treated and the particular method of administration. Preferably, the compositions of the invention should be formulated so that doses of 0.01 to 100 mg/kg body weight/day of the compounds of the invention can be administered.

In compositions that include one or more other therapeutic agent(s), the one or more other therapeutic agent(s) and the compound of the invention may act synergistically. Accordingly, the amount of one or more other therapeutic agent(s) in such compositions may be less than that required in monotherapy with only that therapeutic agent. In such compositions, one or more other therapeutic agent(s) can be administered at a dose of 0.01 to 1,000 μg/kg body weight/day.

The amount of one or more therapeutic agents present in a composition of the present disclosure cannot be greater than the amount that would normally be administered in a composition containing that therapeutic agent as the only active agent. Preferably, the amount of one or more therapeutic agent(s) in the compositions of the present disclosure is about 50% to 100% of the amount normally present in a composition containing that therapeutic agent as the only therapeutically active agent. within the range of In some embodiments, the one or more other therapeutic agent(s) is about 50%, about 55%, about 60%, about 65%, about A dose of 70%, about 75%, about 80%, about 85%, about 90%, or about 95% is administered. As used herein, the phrase "ordinarily administered" means the amount of the FDA-approved therapeutic agent that is approved for administration in accordance with the FDA label insert.

The compounds of the invention, or pharmaceutical compositions thereof, can also be incorporated into compositions for coating implantable medical devices such as prostheses, prosthetic valves, vascular grafts, stents, and catheters. Vascular stents have been used, for example, to overcome restenosis (re-narrowing of the blood vessel wall after injury). However, patients using stents, or other implantable devices, run the risk of clot formation, or platelet activation. These unwanted effects can be prevented or mitigated by precoating the device with a pharmaceutically acceptable composition that includes a kinase inhibitor. Implantable devices coated with compounds of the invention are another embodiment of the invention.

Exemplary Other Therapeutic Agents In some embodiments, the one or more other therapeutic agents are poly ADP ribose polymerase (PARP) inhibitors. In some embodiments, the PARP inhibitor is olaparib (LYNPARZA®, AstraZeneca); rucaparib (RUBRACA®, Clovis Oncology); niraparib (ZEJULA®, Tesaro); talazoparib (MDV3800/ BMN 673/LT00673, Medivation/Pfizer/Biomarin); veliparib (ABT-888, AbbVie); and BGB-290 (BeiGene, Inc.).

In some embodiments, the one or more other therapeutic agents are histone deacetylase (HDAC) inhibitors. In some embodiments, the HDAC inhibitor is vorinostat (ZOLINZA®, Merck); romidepsin (ISTODAX®, Celgene); panobinostat (FARYDAK®, Novartis); belinostat (BELEODAQ®). Entinostat (SNDX-275, Syndax Pharmaceuticals) (NCT00866333); and Chidamide (EPIDAZA®, HBI-8000, Chipscreen B) iosciences, China).

In some embodiments, the one or more other therapeutic agents are CDK inhibitors, such as CDK4/CDK6 inhibitors. In some embodiments, the CDK4/6 inhibitor is palbociclib (Ibrance®, Pfizer); ribociclib (Kisqali®, Novartis); abemaciclib (Ly2835219, Eli Lilly); and tril aciclib(G1T28,G1 Therapeutics).

In some embodiments, the one or more other therapeutic agents are phosphatidylinositol 3-kinase (PI3K) inhibitors. In some embodiments, the PI3K inhibitor is idelalisib (Zydelig®, Gilead), alpelisib (BYL719, Novartis), taselisib (GDC-0032, Genentech/Roche); pictilisib (GDC-0941, Genentech/Roche) he ); copanlisib (BAY806946, Bayer); duvelisib (formerly IPI-145, Infinity Pharmaceuticals); PQR309 (Piqur Therapeutics, Switzerland); and TGR120 2 (formerly RP5230, TG Therapeutics).

In some embodiments, the one or more other therapeutic agents are platinum-based therapeutics, also referred to as platins. Platin causes DNA cross-linking and inhibits DNA repair and/or DNA synthesis primarily in rapidly proliferating cells such as cancer cells.

In some embodiments, the platinum-based therapeutic agent is cisplatin (Platinol®, Bristol-Myers Squibb); carboplatin (Paraplatin®, Bristol-Myers Squibb; Teva; also from Pfizer); oxalipl atin (Eloxitin® Sanofi-Aventis); nedaplatin (Aqupla®, Shionogi), picoplatin (Poniard Pharmaceuticals); and satraplatin (JM-216, Agen nix).

In some embodiments, the one or more other therapeutic agents are taxane compounds, which cause disruption of microtubules that are essential for cell division. In some embodiments, the taxane compound is paclitaxel (Taxol®, Bristol-Myers Squibb), docetaxel (Taxotere®, Sanofi-Aventis; Docefrez®, Sun Pharmaceutical), Bumin-bound paclitaxel (Abraxane®; Abraxis/Celgene), cabazitaxel (Jevtana®, Sanofi-Aventis), and SID530 (SK Chemicals, Co.) (NCT00931008).

In some embodiments, the one or more other therapeutic agents are nucleoside inhibitors or interfere with normal DNA synthesis, protein synthesis, cell replication, or otherwise cause rapid cell proliferation. It is a therapeutic agent that inhibits

In some embodiments, the nucleoside inhibitor is trabectedin (guanidine alkylating agent, Yondelis®, Janssen Oncology), mechlorethamine (alkylating agent, Valchlor®, Aktelion Pharmaceuticals); vincristine (Oncovin®). Trademark), Eli Lilly; Vincasar(R), Teva Pharmaceuticals; Marqibo(R), Talon Therapeutics); Temozolomide (alkylating agent 5-(3-methyltriazen-1-yl)-imidazole-4-carboxamide) Cytarabine Injection (ara-C, an antimetabolite cytidine analog, Pfizer); Lomustine (alkylating agent, CeeNU®, Bristol-Myers Squibb) Gleostine®, NextSource Biotechnology); Azacytidine (pyrimidine nucleoside analog of cytidine, Vidaza®, Celgene); Omacetaxine mepesuccinate (cephalotaxin ester) (protein synthesis inhibitor, Synribo® ); Teva Pharmaceuticals); asparaginase Erwinia chrysanthemi (enzyme for asparagine depletion, Elspar®, Lundbeck; Erwinaze®, EUSA Pharma); eribulin mesylate (microtubule inhibitor, tubulin cell division inhibitor , Halaven®, Eisai); cabazitaxel (microtubule inhibitor, tubulin cell division inhibitor, Jevtana®, Sanofi-Aventis); capacetrin (thymidylate synthase inhibitor, Xeloda®, Bendamustine (bifunctional mechlorethamine derivative, thought to form interstrand DNA cross-links, Treanda®, Cephalon/Teva); Ixabepilone (semi-synthetic analogue of epothilone B, microtubule inhibitor, Tube); Phosphorous cell division inhibitor, Ixempra®, Bristol-Myers Squibb); nelarabine (prodrug of deoxyguanosine analog, nucleoside metabolism inhibitor, Arranon®, Novartis); chlorafabine (ribonucleotide reductase inhibitor) and trifluridine and tipiracil (thymidine-based nucleoside analogs and thymidine phosphorylase inhibitors, Lonsurf®, Taiho Oncology); selected from.

In some embodiments, the one or more other therapeutic agents are kinase inhibitors or VEGF-R antagonists. In some embodiments, the one or more other therapeutic agents are MEK inhibitors. As used herein, "MEK inhibitor" means any inhibitor or blocker or antagonist that binds to and/or inhibits the mitogen-activated protein kinase enzymes MEK1 and/or MEK2. means. In some embodiments, the MEK inhibitor is described by Cheng et al., herein incorporated by reference in its entirety. , “Current Development Status of MEK Inhibitors,” Molecules 2017, 22, 1551. In certain embodiments, the MEK inhibitor is binimetinib (MEK 162, ARRY-438162, ARRAY BIOPHARMA INC.), cobimetinib (COTELLIC®, Exelexis/Genentech/Roche), refametinib (BAY 86-9 766,RDEA119; Bayer AG), selumetinib (AZD6244, ARRY-142886; ASTRAZENECA), trametinib (MEKINIST®, Novartis), mirdametinib (PD-0325901, Spring Works Therapeutic s), pimasertib (AS703026, MSC1936369B, Merck KGaA), or the aforementioned selected from pharmaceutically acceptable salts and/or solvates of any of the following. In certain embodiments, the second anti-cancer agent is binimetinib, cobimetinib, selumetinib, trametinib, mirdametinib, pimasertib, or a pharmaceutically acceptable salt and/or solvate of any of the foregoing. Other examples of MEK inhibitors for use as other therapeutic agents in the methods and uses described herein include E6201 (Eisai Co Ltd./Strategia Therapeutics), 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 second anti-cancer agents in the methods and uses described herein include WO 2005, each of which is incorporated herein by reference in its entirety. Examples include, but are not limited to, those described in WO2021/121142, WO2014/169843, WO2016/035008, WO2016/168704, WO2020/125747, WO2021/142144, WO2021/142345, and WO2021/149776.

In some embodiments, the one or more other therapeutic agents are EGFR inhibitors. As used herein, "EGFR inhibitor" means any inhibitor or blocker or antagonist that binds to and/or inhibits epidermal growth factor receptor (EGFR). In some embodiments, the EGFR inhibitor is described by Ayati et al., herein incorporated by reference in its entirety. , “A review of progression of epidermal growth factor receptor (EGFR) inhibitors as an effective approach in cancer targeted t therapy,” Bioorganic Chemistry 2020, 99: 103811. In some embodiments, the EGFR inhibitor is selected from cetuximab, necitumumab, panitumumab, zaltumumab, nimotuzumab, and matuzumab. In some embodiments, the EGFR inhibitor is cetuximab. In some embodiments, the EGFR inhibitor is necitumumab. In some embodiments, the EGFR inhibitor is panitumumab. In some embodiments, the EGFR inhibitor is zaltumumab. In some embodiments, the EGFR inhibitor is nimotuzumab. In some embodiments, the EGFR inhibitor is matuzumab.

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.

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, effective in the first-line treatment of NSCLC with EGFR-activating mutations, such as deletions in exon 19 and exon 21 L858R mutations. .

In some embodiments, the EGFR inhibitor is a "second generation EGFR tyrosine kinase inhibitor" (second generation TKI). Second generation TKI refers to shared irreversible EGFR inhibitors, such as afatinib and dacomitinib, that are effective in the first-line treatment of NSCLC with EGFR-activating mutations, such as deletions in exon 19 and exon 21 L858R mutations. do.

In some embodiments, the EGFR inhibitor is a "third generation EGFR tyrosine kinase inhibitor" (third generation TKI). Third generation TKIs are selective for EGFR activating mutations, such as deletions in exon 19 and exon 21 L858R, alone or in combination with the T790M mutation, and have low inhibition against wild-type EGFR. refers to covalent irreversible EGFR inhibitors, such as osimertinib and lazertinib, that have activity.

In some embodiments, the one or more other therapeutic agents include the anti-VEGF monoclonal antibody bevacizumab (AVASTIN®, Genentech/Roche); the anti-VEGF antibody ramucirumab (CYRAMZA®, Eli Lilly); ), the anti-VEGFR-2 antibody known as VEGF Trap (ZALTRAP®, Regeneron/Sanofi), and difaflibercept are selected from the approved VEGF inhibitors and kinase inhibitors useful in the present invention. Ru. VEGFR inhibitors, such as regorafenib (STIVARGA®, Bayer); vandetanib (CAPRELSA®, AstraZeneca); axitinib (INLYTA®, Pfizer); and lenvatinib (LENVIMA®, Eisai) ; Raf inhibitors, such as sorafenib (NEXAVAR®, Bayer AG and Onyx); dabrafenib (TAFINLAR®, Novartis); and vemurafenib (ZELBORAF®, Genentech/Roche); MEK inhibitors; For example, cobimetanib (COTELLIC®, Exelexis/Genentech/Roche); trametinib (MEKINIST®, Novartis); Bcr-Abl tyrosine kinase inhibitors, such as imatinib (Gleevec®, Novartis); Bu (TASIGNA®, Novartis); dasatinib (SPRYCEL®, Bristol MyersSquibb); bosutinib (BOSULIF®, Pfizer); and ponatinib (INCLUSIG®, Ariad Pharma) euticals); Her2 and EGFR inhibition agents, such as gefitinib (IRESSA®, AstraZeneca); erlotinib (TARCEEVA®, Genentech/Roche/Astellas); lapatinib (TYKERB®, Novartis); afatinib (GILOTRIF); registered trademark), Boehringer Ingelheim); osimertinib (targets activated EGFR, TAGRISSO®, AstraZeneca); and brigatinib (ALUNBRIG®, Ariad Pharmaceuticals); (COMETRIQ) ®, Exelexis); as well as multi-kinase inhibitors, such as sunitinib (SUTENT®, Pfizer); pazopanib (VOTRIENT®, Novartis); ALK inhibitors, such as crizotinib (XALKORI®); ), Pfizer); ceritinib (ZYKADIA®, Novartis); and alectinib (ALECENZa®, Genentech/Roche); tyrosine kinase inhibitors, e.g. s/Janssen) ; as well as Flt3 receptor inhibitors, such as midostaurin (RYDAPT®, Novartis)

Other kinase inhibitors and VEGF antagonists that are in development and can be used in the present invention include tivozanib (Aveo Pharmaceuticals); batalanib (Bayer/Novartis); lusitanib (Clovis Oncology); dovitinib (TKI258, Novartis); Chiauanib (Chipscreen Biosciences); CEP-11981 (Cephalon); linifanib (Abbott Laboratories); neratinib (HKI-272, Puma Biotechnology); radotinib (Supect®, I Y5511, Il-Yang Pharmaceuticals, S. Korea); ruxolitinib ( Jakafi (registered trademark), Incyte Corporation); PTC299 (PTC Therapeutics); CP-547,632 (Pfizer); foretinib (Exelexis, GlaxoSmithKline); quizartinib (Daiichi Sankyo), and motesanib (Amgen/Takeda).

In some embodiments, the one or more other therapeutic agents are mTOR inhibitors, which inhibit cell proliferation, angiogenesis, and glucose uptake. In some embodiments, the mTOR inhibitors are everolimus (Afinitor®, Novartis); temsirolimus (Torisel®, Pfizer); and sirolimus (Rapamune®, Pfizer).

In some embodiments, the one or more other therapeutic agents are proteasome inhibitors. Approved proteasome inhibitors useful in the present invention include bortezomib (Velcade®, Takeda); carfilzomib (Kyprolis®, Amgen); and ixazomib (Ninlaro®, Takeda). Can be mentioned.

In some embodiments, the one or more other therapeutic agents are growth factor agonists, such as platelet-derived growth factor (PDGF), or antagonists of epidermal growth factor (EGF) or its receptor (EGFR). Approved PDGF antagonists that can be used in the present invention include olaratumab (Lartruvo®; Eli Lilly). Approved EGFR antagonists that can be used in the present invention include cetuximab (Erbitux®, Eli Lilly); necitumumab (Portrazza®, Eli Lilly), panitumumab (Vectibix®, Amgen) ; and osimertinib (targets activated EGFR, Tagrisso®, AstraZeneca).

In some embodiments, the one or more other therapeutic agents are aromatase inhibitors. In some embodiments, the aromatase inhibitor is selected from exemestane (Aromasin®, Pfizer); anastazole (Arimidex®, AstraZeneca), and letrozole (Femara®, Novartis) .

In some embodiments, the one or more other therapeutic agents are hedgehog pathway antagonists. Approved hedgehog pathway inhibitors that can be used in the present invention include sonidegib (Odomzo®, Sun Pharmaceuticals); and vismodegib (Erivedge®), both for treating basal cell carcinoma. Genentech).

In some embodiments, the one or more other therapeutic agents are folate inhibitors. Approved folate inhibitors useful in the present invention include pemetrexed (Alimta®, Eli Lilly).

In some embodiments, the one or more other therapeutic agents are CC chemokine receptor 4 (CCR4) inhibitors. CCR4 inhibitors that have been investigated as potentially useful in the present invention include mogamulizumab (Poteligeo®, Kyowa Hakko Kirin, Japan).

In some embodiments, the one or more other therapeutic agents are isocitrate dehydrogenase (IDH) inhibitors. IDH inhibitors that have been studied for use in the present invention include AG120 (Celgene; NCT02677922); AG221 (Celgene, NCT02677922; NCT02577406); BAY1436032 (Bayer, NCT02746081); IDH305 (No. vartis, NCT02987010) It will be done.

In some embodiments, the one or more other therapeutic agents are arginase inhibitors. Arginase inhibitors that have been studied for use in the present invention include acute myeloid leukemia and myelodysplastic syndromes (NCT02732184) and solid tumors (NCT02561234); and CB-1158 (Calithera Biosciences). One example is AEB1102 (PEGylated recombinant arginase, Aeglea Biotherapeutics), which is being investigated in phase I clinical trials.

In some embodiments, the one or more other therapeutic agents are glutaminase inhibitors. Glutaminase inhibitors that can be used in the present invention include CB-839 (Calithera Biosciences).

In some embodiments, the one or more other therapeutic agents are antibodies that bind to tumor antigens, ie, proteins expressed on the cell surface of tumor cells. Approved antibodies that bind to tumor antigens that can be used in the present invention include rituximab (Rituxan®, Genentech/BiogenIdec); ofatumumab (anti-CD20, Arzerra®, GlaxoSmithKline); obinutuzumab (anti-CD20 , Gazyva®, Genentech), ibritumomab (anti-CD20 and yttrium-90, Zevalin®, Spectrum Pharmaceuticals); daratumumab (anti-CD38, Darzalex®, Janssen Biotech) ), dinutuximab (antiglycolipid GD2, Unituxin®, United Therapeutics); trastuzumab (anti-HER2, Herceptin®, Genentech); ado-trastuzumab emtansine (anti-HER2, fused to emtansine, Kadcyla®, Genentech); and pertuzumab (Anti-HER2 and brentuximab vedotin (anti-CD30-drug conjugate, Adcetris®, Seattle Genetics).

In some embodiments, the one or more other therapeutic agents are topoisomerase inhibitors. Approved topoisomerase inhibitors useful in the present invention include irinotecan (Onivyde®, Merrimack Pharmaceuticals); topotecan (Hycamtin®, GlaxoSmithKline). Topoisomerase inhibitors that have been investigated for use in the present invention include pixantrone (Pixuvri®, CTI Biopharma).

In some embodiments, the one or more other 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 (Venclexta®, AbbVie/Genentech); and blinatumomab (Blincyto®, Amgen). Other therapeutic agents that target apoptotic proteins that are undergoing clinical trials and can be used in the present invention include navitoclax (ABT-263, Abbott), a BCL-2 inhibitor (NCT02079740) .

In some embodiments, the one or more other therapeutic agents are androgen receptor inhibitors. Approved androgen receptor inhibitors useful in the present invention include enzalutamide (Xtandi®, Astellas/Medication); approved inhibitors of androgen synthesis include abiraterone (Zytiga®); ), Centocor/Ortho); approved gonadotropin-releasing hormone (GnRH) receptor antagonists (Degarelix, Firmagon®, Ferring Pharmaceuticals).

In some embodiments, the one or more other therapeutic agents are selective estrogen receptor modulators (SERMs), which interfere with estrogen synthesis or activity. Approved SERMs useful in the present invention include raloxifene (Evista®, Eli Lilly).

In some embodiments, the one or more other therapeutic agents are bone resorption inhibitors. An approved therapeutic agent that inhibits bone resorption is denosumab (Xgeva®, Amgen), which is found on the surface of osteolytic cells, their precursors, and osteoclast-like giant cells. This is an antibody that binds to RANKL and prevents it from binding to its receptor RANK, which mediates bone pathology in solid tumors with bone metastasis. Other approved therapeutic agents that inhibit bone resorption include bisphosphonates such as zoledronic acid (Zometa®, Novartis).

In some embodiments, the one or more other therapeutic agents are inhibitors of the interaction of the two major p53 suppressor proteins, MDMX and MDM2. Inhibitors of p53 suppressor proteins that have been studied to be usable in the present invention include ALRN, a stapled peptide that binds equally to MDMX and MDM2 and equally prevents the interaction of MDMX and MDM2 with p53; -6924 (Aileron). ALRN-6924 is currently being investigated in clinical trials for the treatment of AML, progressive myelodysplastic syndrome (MDS), and peripheral T-cell lymphoma (PTCL) (NCT02909972; NCT02264613).

In some embodiments, the one or more other therapeutic agents are inhibitors of transforming growth factor beta (TGF-beta or TGF-β). Inhibitors of TGF-beta protein that have been investigated for use in the present invention include various cancers including breast cancer, lung cancer, hepatocellular carcinoma, colon cancer, pancreatic cancer, prostate cancer, and renal cancer (NCT 02947165). NIS793 (Novartis) is an anti-TGF-beta antibody that is in clinical trials for the treatment of cancer. In some embodiments, the inhibitor of TGF-beta protein is fresolimumab (GC1008; Sanofi-Genzyme), which is used to treat melanoma (NCT00923169); renal cell carcinoma (NCT00356460); and non-small cell lung cancer (NCT02581787). ) has been studied. Furthermore, in some embodiments, the additional therapeutic agent is described by Connolly et al. (2012) Int’l J. TGF-beta trap, as described in Biological Sciences 8:964-978. Certain therapeutic compounds currently in clinical trials for the treatment of solid tumors are the bispecific anti-PD-L1/TGF-β trap compound (NCT02699515); and (NCT02517398), M7824 ( Merck KgaA-formerly MSB0011459X). M7824 is composed of a fully human IgG1 antibody directed against PD-L1 fused to the extracellular domain of human TGF-beta receptor II, which functions as a TGF-β "trap."

In some embodiments, the one or more other therapeutic agents are glembatumumab vedotin-monomethyl auristatin E (MMAE), which is an anti-glycoprotein NMB (gpNMB) antibody (CR011) conjugated to the cytotoxic MMAE. ) (Celldex). gpNMB is a protein overexpressed by multiple tumor types that is associated with the ability of cancer cells to metastasize.

In some embodiments, the one or more other therapeutic agents are anti-proliferative compounds. Such antiproliferative compounds include aromatase inhibitors; antiestrogens; topoisomerase I inhibitors; topoisomerase II inhibitors; microtubule-activating compounds; alkylating compounds; histone deacetylase inhibitors; inducing cell differentiation processes. cyclooxygenase inhibitors; MMP inhibitors; mTOR inhibitors; antineoplastic antimetabolites; platin compounds; compounds that target/reduce protein or lipid kinase activity; and further anti-angiogenic compounds; proteins or lipids. Compounds that target, reduce, or inhibit the activity of phosphatases; gonadorelin agonists; antiandrogens; methionine aminopeptidase inhibitors; matrix metalloproteinase inhibitors; bisphosphonates; biological response modifiers; antiproliferative antibodies; heparanase Inhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasome inhibitors; compounds for use in the treatment of hematological malignancies; compounds that target, reduce, or inhibit the activity of Flt-3; manufactured by Conforma Therapeutics such as 17-AAG (17-allylaminogeldamycin, NSC330507), 17-DMAG (17-dimethylaminoethylamino-17-methoxy-geldamycin, NSC707545), IPI-504, CNF1010, CNF2024, CNF1010. Hsp inhibitors; temozolomide (TEMODAL®); kinesin spindle protein inhibitors such as SB715992 or SB743921 from GlaxoSmithKline or pentamidine/chlorpromazine from CombinatoRx; ARRY142 from Array BioPharma 886, AZd ₆ 244 manufactured by AstraZeneca , PD181461 from Pfizer, and leucovorin.

As used herein, the term "aromatase inhibitor" relates to compounds that inhibit estrogen production, such as the conversion of the substrates androstenedione and testosterone to estrone and estradiol, respectively. The term includes steroids, especially atamestane, exemestane, and formethane, and especially non-steroids, especially aminoglutethimide, logrethymide, pyridoglutethimide, trilostane, testolactone, ketoconazole, vorozole, fadrozole, anastrozole, and letrozole, but are not limited to these. Exemestane is sold under the trade name Aromasin™. Formethane is sold under the trade name Lentaron(TM). Fadrozole is sold under the trade name Afema(TM). Anastrozole is sold under the trade name Arimidex™. Letrozole is sold under the trade name Femara(TM) or Femar(TM). Aminoglutethimide is sold under the trade name Orimeten(TM). Combinations of the invention comprising chemotherapeutic agents that are aromatase inhibitors are particularly useful in the treatment of hormone receptor positive tumors such as breast tumors.

As used herein, the term "antiestrogen" refers to a compound that antagonizes the effects of estrogen at the estrogen receptor level. This term includes, but is not limited to, tamoxifen, fulvestrant, raloxifene, and raloxifene hydrochloride. Tamoxifen is sold under the trade name Nolvadex(TM). Raloxifene hydrochloride is sold under the trade name Evista™. Fulvestrant can be administered under the trade name Faslodex™. Fulvestrant can be administered under the trade name Faslodex™. Combinations of the invention comprising chemotherapeutic agents that are antiestrogen agents are particularly useful in the treatment of estrogen receptor positive tumors, such as breast tumors.

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

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

As used herein, the term "topoisomerase II inhibitor" refers to anthracyclines, such as doxorubicin (including liposomal formulations such as Caelyx™), daunorubicin, epirubicin, idarubicin, and nemorubicin; xantrone and rosoxantrone; and the podophyllotoxins etoposide and teniposide. Etoposide is sold under the trade name Etophos™. Teniposide is sold under the trade name VM 26-Bristol. Doxorubicin is sold under the trade names Acriblastin™ or Adriamycin™. Epirubicin is sold under the trade name Farmorubicin(TM). Idarubicin is sold under the trade name Zavedos™. Mitoxantrone is sold under the trade name Novantron(TM).

The term "microtubule activator" includes, but is not limited to, taxanes, such as paclitaxel and docetaxel; vinca alkaloids, such as vinblastine or vinblastine sulfate, vincristine or vincristine sulfate, and vinorelbine; discodermolide; The present invention relates to microtubule stabilizing and microtubule destabilizing compounds, and polymerization inhibitors, including colchicine and epothilone, and derivatives thereof. Paclitaxel is sold under the trade name Taxol(TM). Docetaxel is sold under the trade name Taxotere(TM). Vinblastine sulfate is available from Vinblastin R. It is sold under the trade name P (trademark). Vincristine sulfate is sold under the trade name Farmistin™.

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

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

The term "anti-tumor antimetabolites" includes 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. However, it is not limited to these. Capecitabine is sold under the trade name Xeloda™. Gemcitabine is sold under the trade name Gemzar(TM).

As used herein, the term "platin compound" includes, but is not limited to, carboplatin, cisplatin, cisplatinum, and oxaliplatin. Carboplatin can be administered, for example, in the form as sold under the trade name Carboplat(TM). Oxaliplatin can be administered, for example, in the form as it is sold under the tradename Eloxatin™.

As used herein, the term "a compound that targets/reduces protein or lipid kinase activity; or protein or lipid phosphatase activity; or further anti-angiogenic compound" refers to protein tyrosine kinases and/or serine and / or threonine kinase inhibitors or lipid kinase inhibitors, e.g. a) compounds that target, reduce or inhibit the activity of platelet-derived growth factor receptor (PDGFR), e.g. Compounds that reduce or inhibit, in particular compounds that inhibit PDGF receptors, such as N-phenyl-2-pyrimidine-amine derivatives, such as imatinib, SU101, SU6668, and GFB-111; b) fibroblast cell growth c) compounds that target, reduce or inhibit the activity of insulin-like growth factor receptor I (IGF-IR); For example, compounds that target, reduce, or inhibit the activity of IGF-IR, in particular compounds that inhibit the kinase activity of the IGF-I receptor, or the extracellular domain of the IGF-I receptor or its growth factors. d) a compound that targets, reduces, or inhibits the activity of the Trk receptor tyrosine kinase family, or an ephrinB4 inhibitor; e) a compound that targets the activity of the AxI receptor tyrosine kinase family; f) a compound that targets, reduces or inhibits the activity of a Ret receptor tyrosine kinase; g) a compound that targets, reduces or inhibits the activity of a Kit/SCFR receptor tyrosine kinase; h) a compound that targets, reduces or inhibits the activity of a C-kit receptor tyrosine kinase, which is part of the PDGFR family, such as a member of the c-kit receptor tyrosine kinase family; Compounds that target, reduce or inhibit the activity of the c-kit receptor, in particular compounds that inhibit the c-kit receptor, such as imatinib; i) members of the c-Abl family, gene fusion products thereof, such as BCR-Abl kinase; ), such as compounds that target, reduce or inhibit the activity of c-Abl family members and their gene fusion products, such as N-phenyl- 2-pyrimidine-amine derivatives, such as imatinib or nilotinib (AMN107) from Parke Davis; PD180970; AG957; NSC680410; PD173955; or dasatinib (BMS-354825); Members of the kinase family MEK, SRC, JAK/pan-JAK, FAK, PDK1, PKB/Akt, Ras/MAPK, PI3K, SYK, TYK2, BTK, and TEC families, and/or staurosporine derivatives. Compounds that target, reduce, or inhibit the activity of members of the cyclin-dependent kinase family (CDK), such as midostaurin; (additional examples of compounds include UCN-01, Safingol, BAY 43-9006, bryostatin) 1, perifosine; irmofosine; RO 318220 and RO 320432; GO 6976; lsis 3521; LY333531/LY379196; isoquinoline compounds; FTIs; ); k) compounds that target, reduce or inhibit the activity of protein-tyrosine kinase inhibitors, such as imatinib mesylate (Gleevec™) or tyrphosine, such as Tyrphostin A23/RG-50810; AG 99 ;Tyrphostin AG 213;Tyrphostin AG 1748;Tyrphostin AG 490;Tyrphostin B44;Tyrphostin B44 (+) enantiomer;Tyrphostin AG 555;AG 494;Tyrph ostin AG 556, AG957, and adafosine (4-{[(2,5-dihydrophenyl ) methyl]amino}-benzoic acid adamantyl ester; NSC 680410, adaphostin); l) receptor tyrosine of the epidermal growth factor family; Compounds that target, reduce or inhibit the activity of kinases (EGFR1, ErbB2, ErbB3, ErbB4 as homo- or heterodimers) and variants thereof, e.g. Compounds that target, reduce, or inhibit, particularly compounds that inhibit members of the EGF receptor tyrosine kinase family, such as EGF receptors, ErbB2, ErbB3, and ErbB4, or that bind to EGF or EGF-related ligands. , protein, or antibody, i.e. CP 358774, ZD 1839, ZM 105180; trastuzumab (Herceptin™), cetuximab (Erbitux™), Iressa, Tarceva, OSI-774, Cl-1033, EKB-569, GW -2016, E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3 or E7.6.3, and 7H-pyrrolo-[2,3-d]pyrimidine derivatives; m) compounds that target, reduce or inhibit the activity of c-Met receptors, such as compounds that target, reduce or inhibit the activity of c-Met, in particular c-Met receptors; Compounds that inhibit the body's kinase activity, or antibodies that target the extracellular domain of c-Met or bind to HGF, n) PRT-062070, SB-1578, Baricitinib, Pacritinib, Momelotinib, VX-509, Targeting the kinase activity of one or more JAK family members (JAK1/JAK2/JAK3/TYK2 and/or pan-JAKs), including but not limited to AZD-1480, TG-101348, tofacitinib, and ruxolitinib , reducing or inhibiting compounds; o) ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, Buparisib, Pictrellisib, PF-4691502, BYL-719, Dactolisib, XL -147, XL-765, and idelalisib, and compounds that target, reduce, or inhibit PI3 kinase (PI3K); and; and q) cyclopamine, vismodegib, itraconazole, erismodegib, Compounds that target, reduce, or inhibit the signaling effects of the hedgehog protein (Hh) or smoothed receptor (SMO) pathway, including but not limited to IPI-926 (saridegib); but not limited to.

As used herein, the term "PI3K inhibitor" refers 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. including, but not limited to, compounds with Examples of PI3K inhibitors useful in the present invention include ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, Buparisib, Pictrellisib, PF-4691502, BYL-719, Dactolisib. , XL-147, XL-765, and idelalisib.

As used herein, the term "Bcl-2 inhibitor" refers to ABT-199, ABT-731, ABT-737, apogossypol, the pan-Bcl-2 inhibitor from Ascenta, curcumin (and analogs thereof). ), dual Bcl-2/Bcl-xL inhibitor (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 analogues, see WO2004106328), S-001 (Gloria Pharmaceuticals), TW series of compounds (U niv. of Michigan), and Compounds that have inhibitory activity against B-cell lymphoma 2 protein (Bcl-2) include, but are not limited to, venetoclax. In some embodiments, the Bcl-2 inhibitor is a small molecule therapeutic. In some embodiments, the Bcl-2 inhibitor is a peptidomimetic.

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

As used herein, the term "SYK inhibitor" refers to spleen tyrosine kinase (SYK ), including, but not limited to, compounds having inhibitory activity against.

BTK inhibitory compounds and medical conditions treatable by such compounds in combination with compounds of the invention can be found in WO2008039218 and WO2011090760, which are incorporated herein by reference in their entirety.

SYK inhibitory compounds and medical conditions treatable by such compounds in combination with compounds of the invention can be found in WO2003063794, WO2005007623, and WO2006078846, which are incorporated herein by reference in their entirety. Can be done.

PI3K inhibitory compounds and conditions treatable by such compounds in combination with compounds of the present invention include WO2004019973, WO2004089925, WO2007016176, US8138347, WO2002088112, which are incorporated herein by reference in their entirety. , WO2007084786, WO2007129161, WO2006122806, WO2005113554, and WO2007044729.

JAK inhibitory compounds and conditions treatable by such compounds in combination with the compounds of the present invention are described in WO2009114512, WO2008109943, WO2007053452, WO2000142246, and It can be found in WO2007070514.

Additional anti-angiogenic compounds include compounds that have other mechanisms of activity, such as those unrelated to protein or lipid kinase inhibition, such as Thalomid™ and TNP-470.

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

Compounds that target, reduce or inhibit the activity of protein or lipid phosphatases are, for example, inhibitors of phosphatase 1, phosphatase 2A, or CDC25, such as okadaic acid or its derivatives.

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.

As used herein, the term "cyclooxygenase inhibitor" refers to Cox-2 inhibitors, 5-alkyl substituted 2-arylaminophenyl acetic acids and derivatives, such as celecoxib (

Celebrex™), rofecoxib (Vioxx™), etoricoxib, valdecoxib, or 5-alkyl-2-arylaminophenyl acetic acid, such as 5-methyl-2-(2'-chloro-6'-fluoroanidine). including, but not limited to, lino)phenylacetic acid, lumiracoxib.

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

As used herein, the term "heparanase inhibitor" refers to a compound that targets, reduces, or inhibits heparin sulfate degradation. This term includes, but is not limited to, PI-88. As used herein, the term "biological response modifier" refers to lymphokines or interferons.

As used herein, the term "inhibitor of a Ras oncogenic isoform," such as H-Ras, K-Ras, or N-Ras, refers to targeting, reducing, or inhibiting the oncogenic activity of Ras. a farnesyltransferase inhibitor such as L-744832, DK8G557, or R115777 (Zarnestra™). As used herein, the term "telomerase inhibitor" refers to a compound that targets, reduces, or inhibits the activity of telomerase. Compounds that target, reduce or inhibit the activity of telomerase are in particular compounds that inhibit telomerase receptors, such as telomestatin.

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

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 proteasome activity include, but are not limited to, bortezomib (Velcade™) and MLN341.

As used herein, the term "matrix metalloproteinase inhibitor", or ("MMP" inhibitor), refers to collagen peptidomimetic and non-peptide mimetic inhibitors, tetracycline derivatives, e.g., hydroxamate peptidomimetics. batimastat, and its orally administrable analogues marimastat (BB-2516), prinomastat (AG3340), metastat (NSC 683551), BMS-279251, BAY 12-9566, TAA211, MMI270B, or AAJ996, but are not limited to these.

As used herein, the term "compound for use in the treatment of hematological malignancies" is a compound that targets, reduces, or inhibits the activity of the FMS-like tyrosine kinase receptor (Flt-3R). FMS-like tyrosine kinase inhibitors; interferon, 1-β-D-arabinofuranosylcytosine (ara-c) and bisulfan; and ALK, a compound that targets, reduces, or inhibits anaplastic lymphoma kinases. Including, but not limited to, inhibitors.

Compounds that target, reduce, or inhibit the activity of the FMS-like tyrosine kinase receptor (Flt-3R) include, in particular, compounds, proteins, or antibodies that inhibit members of the Flt-3R receptor kinase family, e.g. PKC412, midostaurin, staurosporine derivatives, SU11248, and MLN518.

As used herein, the term "HSP90 inhibitor" targets, reduces, or inhibits the intrinsic ATPase activity of HSP90; degrades, targets HSP90 clients through the ubiquitin proteosome pathway; including, but not limited to, compounds that reduce or inhibit. 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-demethoxygeldanamycin. (17AAG), a geldanamycin derivative; other geldanamycin related compounds: radicicol and HDAC inhibitors.

As used herein, the term "anti-proliferatively active antibody" refers to trastuzumab (Herceptin(TM)), trastuzumab-DM1, erbitux, bevacizumab (Avastin(TM)), rituximab (Rituxan(R)), Including, but not limited to, PRO64553 (anti-CD40), and 2C4 antibody. By antibody is meant intact monoclonal, polyclonal, multispecific antibodies formed from at least two intact antibodies and from antibody fragments so long as they exhibit the desired biological activity.

For the treatment of acute myeloid leukemia (AML), the compounds of the invention can be used in combination with standard leukemia therapies, particularly in combination with therapies used for the treatment of AML. In particular, the compounds of the invention may be combined with, for example, farnesyltransferase inhibitors and/or other drugs useful in the treatment of AML, such as daunorubicin, adriamycin, Ara-C, VP-16, teniposide, mitoxantrone, idarubicin. , carboplatinum, and PKC412.

Other antileukemic compounds include, for example, the pyrimidine analog Ara-C, which is a 2'-α-hydroxyribose (arabinoside) derivative of deoxycytidine. Also included are the purine analogs of 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 hydroxamic acid suberoylanilide (SAHA), are known as histone deacetylases. Inhibits enzyme activity. Specific HDAC inhibitors include MS275, SAHA, FK228 (formerly FR901228), trichostatin A; and N-hydroxy-3-[4-[[[2-(2-methyl-1H-indole-3 -yl)-ethyl]-amino]methyl]phenyl]-2E-2-propenamide, or a pharmaceutically acceptable salt thereof, and N-hydroxy-3-[4-[(2-hydroxyethyl) 2-(1H-indol-3-yl)ethyl]-amino]methyl]phenyl]-2E-2-propenamide, or a pharmaceutically acceptable salt thereof, especially the lactate, US 6,552,065. As used herein, somatostatin receptor antagonist refers to compounds that target, reduce, or inhibit somatostatin receptors, such as octreotide and SOM230. By "approaches that damage tumor cells" is meant approaches such as ionizing radiation. The term "ionizing radiation" as referred to hereinbefore and hereinafter means ionizing radiation that occurs either as electromagnetic radiation (such as X-rays and gamma rays) or as particles (such as alpha and beta particles). Ionizing radiation is provided in, but not limited to, radiation therapy and is known in the art. Hellman, Principles of Radiation Therapy, Cancer, in Principles and Practice of Oncology, Devita et al. , Eds. , ^4th Edition, Vol. 1, pp. 248-275 (1993).

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

In particular, 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine or a pharmaceutically acceptable salt thereof, 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine succinate. ; Angiostatin™; Endostatin™; anthranilic acid amide; ZD4190; Zd ₆ 474; SU5416; SU6668; bevacizumab; ugon etc. ); Also included are compounds, proteins, or monoclonal antibodies of VEGF, such as FLT-4 inhibitors, FLT-3 inhibitors, VEGFR-2 IgGI antibodies, Angiozyme (RPI 4610), and Bevacizumab (Avastin™).

As used herein, "photodynamic therapy" refers to treatments that use certain chemicals known as photosensitive compounds to treat or prevent cancer. Examples of photodynamic therapy include treatment with compounds such as Visudyne™ and porfimer sodium.

As used herein, anti-angiogenic steroids include, for example, anecortave, triamcinolone, hydrocortisone, 11-α-epihydrocotizole, cortexolone, 17α-hydroxyprogesterone, corticosterone, deoxycorticosterone, testosterone, refers to compounds that block or inhibit angiogenesis, such as estrone and dexamethasone.

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

Other chemotherapeutic compounds include plant alkaloids, hormonal compounds, and antagonists; biological response modifiers, preferably lymphokines or interferons; antisense oligonucleotides or oligonucleotide derivatives; shRNA or siRNA; other or unknown mechanisms of action. including, but not limited to, a variety of miscellaneous compound(s) having .

The structure of the active compound, identified by code number, gene name or trade name, can be obtained from the standard compendium "The Merck Index" or from a database, e.g. Patents International (e.g. IMS World Publications). .

Exemplary Immuno-Oncology Agents In some embodiments, the one or more other therapeutic agents are immuno-oncology agents. As used herein, the term "immuno-oncologic agent" refers to an agent effective to enhance, stimulate, and/or upregulate an immune response in a subject. In some embodiments, administering an immuno-oncology agent with a compound of the invention has a synergistic effect in treating cancer.

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

In some embodiments, the immuno-oncology agent is (i) an agonist of a stimulatory (including costimulatory) receptor, or (ii) an agonist of an inhibitory (including co-inhibitory) signal on a T cell. antagonists, and together they result in amplification of antigen-specific T cell responses.

Certain stimulatory and inhibitory molecules are members of the immunoglobulin superfamily (IgSF). One important family of membrane-bound ligands that bind to costimulatory or co-inhibitory receptors is the B7 family, which includes B7-1, B7-2, B7-H1 (PD-L1), Examples include B7-DC (PD-L2), B7-H2 (ICOS-L), B7-H3, B7-H4, B7-H5 (VISTA), and B7-H6. Another family of membrane-bound ligands that bind to costimulatory, or co-inhibitory, receptors is the TNF family of molecules that bind to cognate TNF receptor family members, including CD40 and CD40L, OX-40, OX-40L, CD70, CD27L, CD30, CD30L, 4-1BBL, CD137 (4-1BB), TRAIL/Apo2-L, TRAILR1/DR4, TRAILR2/DR5, TRAILR3, TRAILR4, OPG, 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 , Examples include TNFα, LTβR, lymphotoxin α1β2, FAS, FASL, RELT, DR6, TROY, and NGFR.

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 Cytokines that stimulate T cell activation to stimulate a response.

In some embodiments, a compound of the invention can be combined with an immuno-oncology agent to stimulate a T cell response. In some embodiments, the immuno-oncology agent is (i) an antagonist of a protein that inhibits T cell activation (e.g., an immune checkpoint inhibitor), e.g., 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, They are OX40, OX40L, GITR, GITRL, CD70, CD27, CD40, DR3, and CD28H.

In some embodiments, the immuno-oncology agent is an antagonist of an inhibitory receptor on a NK cell or an agonist of an activating receptor on a NK cell. In some embodiments, the immuno-oncology agent is an antagonist of KIR, such as rililumab.

In some embodiments, the immuno-oncology agent is RG7155 (WO11/70024, WO11/107553, WO11/131407, WO13/87699, WO13/119716, WO13/132044), or FPA-008 (WO11/140249; WO13169264; agents that inhibit or deplete macrophages or monocytes, including, but not limited to, CSF-1R antagonists, such as CSF-1R antagonist antibodies including WO 14/036357).

In some embodiments, the immuno-oncology agent includes one or more of an agonist agent that ligates positive co-stimulatory receptors, an inhibitory receptor, an antagonist, and an agent that increases the frequency of anti-tumor T cells systemically. Blocking agents that attenuate signaling within the tumor microenvironment by overcoming intrinsic immunosuppressive pathways (e.g., blocking the engagement of inhibitory receptors (e.g., PD-L1/PD-1 interaction), (e.g., , depleting or inhibiting Tregs (using anti-CD25 monoclonal antibodies (e.g., daclizumab) or by depletion of anti-CD25 beads ex vivo), inhibiting metabolic enzymes such as IDO, or reversing T cell energy or exhaustion. agents that cause innate immune activation and/or inflammation at the tumor site.

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.

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 cell 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 composed of the extracellular domain of PD-L2 (B7-DC) fused to the Fc portion of IgG1, called AMP-224.

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 antibodies are MPDL3280A (RG7446; WO2010/077634), durvalumab (MEDI4736), BMS-936559 (WO2007/005874), and MSB0010718C (WO2013/79174).

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).

In some embodiments, the immuno-oncology agent is a CD137 (4-1BB) agonist. In some embodiments, the CD137(4-1BB) agonist is an agonistic CD137 antibody. In some embodiments, the CD137 antibody is urelumab or PF-05082566 (WO 12/32433).

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

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

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

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).

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

In some embodiments, the immuno-oncology agent is a CD27 agonist. In some embodiments, the CD27 agonist is an agonist CD27 antibody. In some embodiments, the CD27 antibody is varlilumumab.

In some embodiments, the immuno-oncology agent is MGA271 (against B7H3) (WO 11/109400).

In some embodiments, the immuno-oncology agent is avagovomab, adecatumumab, aftuzumab, alemtuzumab, anatumomab mafenatox, apolizumab, atezolimab, avelumab, blinatumomab, BMS-936559, catumaxomab, durvalumab, epacadostat, epratuzumab, indoximod, inno Tuzumab ozogamicin, intelumab, ipilimumab, isatuximab, lambrolizumab, MED14736, MPDL3280A, nivolumab, obinutuzumab, ocaratuzumab, ofatumumab, oratatuzumab, pembrolizumab, pidilizumab, rituximab, ticilimumab, samalizumab, or tremelimumab.

In some embodiments, the immuno-oncology agent is an immunostimulatory agent. For example, antibodies that block the PD-1 and PD-L1 inhibitory axes can liberate activated tumor-reactive T cells, many of which have not been previously thought to be sensitizing in immunotherapy. It has been shown in clinical trials to induce durable anti-tumor responses in a growing number of tumor histologies, including both tumor types. For example, Okazaki, T. et al. (2013) Nat. Immunol. 14, 1212-1218; Zou et al. (2016) Sci. Transl. Med. Please refer to 8. Nivolumab (Opdivo®, Bristol-Myers Squibb; also known as ONO-4538, MDX1106, and BMS-936558), an anti-PD-1 antibody, is used to treat disease during or before or after anti-angiogenic therapy. has shown the ability to improve overall survival in patients with RCC who experience progression.

In some embodiments, the immunomodulatory therapeutic specifically induces apoptosis of tumor cells. Approved immunomodulatory therapeutics that can be used in the present invention include pomalidomide (Pomalyst®, Celgene); lenalidomide (Revlimid®, Celgene); ingenol mebutate (Picato®). ), LEO Pharma).

In some embodiments, the immuno-oncology agent is a cancer vaccine. In some embodiments, the cancer vaccine is Provexel-T (PROVENGE), which is approved for the treatment of asymptomatic or minimally symptomatic metastatic castration-resistant (hormone-refractory) prostate cancer. (R), Dendreon/Valeant Pharmaceuticals); and Talimo, a recombinant oncolytic viral therapy approved for the treatment of unresectable cutaneous, subcutaneous, and nodular lesions of melanoma. Genla Herparepvec (IMLYGIC®, BioVex/Amgen, formerly known as T-VEC). In some embodiments, the immuno-oncology agent is directed against oncolytic viral therapies, e.g., hepatocellular carcinoma (NCT02562755) and melanoma (NCT00429312), with thymidine recombinant to express GM-CSF Pexastimogendevacilepvec (PexaVec/JX-594), a kinase (TK-)-deficient vaccinia virus; colorectal cancer (NCT01622543); prostate cancer (NCT01619813); head and neck squamous cell carcinoma (NCT01166542); In many cancers, including pancreatic adenocarcinoma (NCT00998322); and non-small cell lung cancer (NSCLC) (NCT 00861627), respiratory intestinal orphan viruses (reoviruses) that do not replicate in cells without RAS activation A variant, Reolysin® (Oncolytics Biotech); recombined to express full-length CD80 and antibody fragments specific for the T-cell receptor CD3 protein in ovarian cancer (NCT02028117). Enadenotskilev (NG-348, PsiOxus, formerly known as ColoAd1), an adenovirus that has been shown to be metastatic or progressive in cancers such as colorectal cancer, bladder cancer, head and neck squamous cell carcinoma, and salivary gland cancer (NCT02636036) ONCOS-102 (Targovax/formerly Oncos), an adenovirus engineered to express GM-CSF, in epithelial tumors; melanoma (NCT03003676); and peritoneal disease, colorectal cancer, or ovarian cancer (NCT02963831). ); Peritoneal Carcinoma (NCT01443260): Fallopian Tube Cancer, Ovarian Cancer (NCT 02759588); GL-ONC1 (GLV-1h68/GLV-1h153, Genelux GmbH), a vaccinia virus engineered to express human sodium iodide symporter (hNIS); expresses GM-CSF in bladder cancer (NCT02365818) selected from recombinant adenoviruses.

In some embodiments, the immuno-oncology agent is a TK and variola growth factor deficient protein that has been engineered to express a cytosine deaminase capable of converting the prodrug 5-fluorocytosine to the cytotoxic drug 5-fluorouracil. JX-929 (SillaJen/formerly Jennerex Biotherapeutics), a vaccinia virus; TG01 and TG02 (Targovax/formerly Oncos), peptide-based immunotherapeutics targeting difficult-to-treat RAS mutations; and Ad5/3- A recombinant adenovirus designated E2F-δ24-hTNFα-IRES-hIL20, TILT-123 (TILT Biotherapeutics); VSV-GP (ViraTherapeutics), a vesicular stomatitis virus (VSV) that has been engineered to express antigens designed to increase antigen-specific CD8 ⁺ T cell responses. Can be recombined.

In some embodiments, the immuno-oncology agent is a T cell that has been engineered to express a chimeric antigen receptor, or CAR. T cells engineered to express such chimeric antigen receptors are called CAR-T cells.

Natural ligands, specific for cell surface antigens fused to the endodomain, the functional terminus of the T cell receptor (TCR), such as the TCR-derived CD3ζ signaling domain, which can generate activation signals in T lymphocytes. CARs have been constructed that are composed of binding domains that can be derived from single chain variable fragments (scFv) derived from standard monoclonal antibodies. Upon antigen binding, such CARs bind to endogenous signaling pathways within effector cells and generate activation signals similar to those initiated by TCR complexes.

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

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

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

Other immuno-oncological agents that can be used in the present invention include urelumab (BMS-663513, Bristol-Myers Squibb), which is an anti-CD137 monoclonal antibody; varlilumab (CDX-1127, Celldex Therapeutics), which is an anti-CD27 monoclonal antibody; Anti-OX40 monoclonal antibody, BMS-986178 (Bristol-Myers Squibb); anti-KIR monoclonal antibody, rililumab (IPH2102/BMS-986015, Innate Pharma, Bristol-Myers Squibb); anti-NKG 2A monoclonal antibody, monalizumab ( IPH2201, Innate Pharma, AstraZeneca); anti-MMP9 antibody, Andecaliximab (GS-5745, Gilead Sciences); and anti-GITR monoclonal antibody, MK-4166 (Merck & Co.).

In some embodiments, the immunostimulatory agent is selected from elotuzumab, mifamurtide, an agonist or activator of toll-like receptors, and an activator of RORγt.

In some embodiments, the immunostimulatory therapeutic agent is recombinant human interleukin 15 (rhIL-15). rhIL-15 is in clinical trials as a treatment 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, IL-15-based immunotherapeutic agents are being tested in Phase I clinical trials against melanoma, renal cell carcinoma, non-small cell lung cancer, and head and neck squamous cell carcinoma (NCT02452268). , a fusion complex composed of a synthetic form of endogenous IL-15 (IL15:sIL-15RA) complexed with the soluble IL-15 binding protein IL-15 receptor alpha chain, a heterodimeric IL. -15 (hetIL-15, Novartis/Admune). In some embodiments, the recombinant human interleukin 12 (rhIL-12) is NM-IL-12 (Neumedicines, Inc.), NCT02544724, or NCT02542124.

In some embodiments, the immuno-oncology agent is described by Jerry L. Adams et al. , “Big opportunities for small molecules in immuno-oncology,” Cancer Therapy 2015, Vol. 14, pages 603-622. In some embodiments, the immuno-oncology agent is manufactured by Jerry L. Adams et al. selected from the examples listed in Table 1. In some embodiments, the immuno-oncology agent is manufactured by Jerry L. Adams et al. A small molecule targeting an immune tumor target selected from those listed in Table 2 of . In some embodiments, the immuno-oncology agent is manufactured by Jerry L. Adams et al. A low molecular weight agent selected from those listed in Table 2.

In some embodiments, the immuno-oncology agent is described by Peter L. Toogood, “Small molecule immuno-oncology therapeutic agents,” Bioorganic & Medicinal Chemistry Letters 2018, Vol. 28 (pages 319-329). In some embodiments, the immuno-oncology agent is described by Peter L. It is an agent that targets the pathway described in Toogood.

In some embodiments, the immuno-oncology agent is described by Sandra L. Ross et al. , “Bispecific T cell engager (BiTE®) antibody constructs can mediate bystander tumor cell killing”, PLoS ONE 12(8): e0183390 Selected from those listed. In some embodiments, the immuno-oncology agent is a bispecific T cell engager (BiTE®) antibody construct. In some embodiments, the bispecific T cell engager (BiTE®) antibody construct is a CD19/CD3 bispecific antibody construct. In some embodiments, the bispecific T cell engager (BiTE®) antibody construct is an EGFR/CD3 bispecific antibody construct. In some embodiments, the bispecific T cell engager (BiTE®) antibody construct activates T cells. In some embodiments, the bispecific T cell engager (BiTE®) antibody construct activates T cells, thereby promoting intercellular adhesion molecule 1 (ICAM-1) in bystander cells. and release cytokines that induce upregulation of FAS. In some embodiments, a bispecific T cell engager (BiTE®) antibody construct activates T cells, which results in lysis of induced bystander cells. In some embodiments, the bystander cells are within a solid tumor. In some embodiments, the bystander cells that are lysed are in close proximity to the BiTE® 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 ex vivo expanded tumor-infiltrating T cells. In some embodiments, the immuno-oncology agent is a bispecific antibody construct or chimeric antigen receptor (CAR) that directly connects T cells with a tumor-associated surface antigen (TAA).

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

As used herein, the term "checkpoint inhibitor" refers to an agent useful in preventing cancer cells from evading a patient's immune system. One of the main mechanisms of destruction of anti-tumor immunity is known as "T cell depletion", which results in long-term exposure to antigen that results in upregulation of inhibitory receptors. These inhibitory receptors function as immune checkpoints to prevent uncontrolled immune responses.

PD-1 and co-inhibitory receptors, such as cytotoxic T lymphocyte antigen 4 (CTLA-4, B and T lymphocyte attenuator (BTLA; CD272), T cell immunoglobulin and mucin domain-3 (Tim- 3), lymphocyte activation gene-3 (Lag-3; CD223), and others are often called checkpoint regulators. It acts as a molecular "gatekeeper" that determines whether or not internal signaling processes proceed.

In some embodiments, the immune checkpoint inhibitor is an antibody against PD-1. PD-1 binds to the programmed cell death 1 receptor (PD-1) and prevents the receptor from binding to the inhibitory ligand PDL-1, thereby allowing tumors to suppress the host's antitumor immune response. disable the ability to

In one aspect, the checkpoint inhibitor is a biological therapeutic or small molecule. In another aspect, the checkpoint inhibitor is a monoclonal antibody, humanized antibody, fully human antibody, fusion protein, or a combination thereof. In a further aspect, the checkpoint inhibitor is CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, Inhibits a checkpoint protein selected from CHK 1, CHK2, A2aR, B-7 family ligands, or a combination thereof. In a further aspect, the checkpoint inhibitor is CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, Interacts with a checkpoint protein ligand selected from CHK 1, CHK2, A2aR, B-7 family ligands, or a combination thereof. In one aspect, the checkpoint inhibitor is an immunostimulant, a T cell growth factor, an interleukin, an antibody, a vaccine, or a combination thereof. In further embodiments, the interleukin is IL-7 or IL-15. In certain embodiments, the interleukin is glycosylated IL-7. In a further aspect, the vaccine is a dendritic cell (DC) vaccine.

Checkpoint inhibitors include any agent that blocks or inhibits the immune system's inhibitory pathways in a statistically significant manner. Such inhibitors can include small molecule inhibitors, or antibodies or antigen-binding fragments thereof that bind to, block or inhibit immune checkpoint receptors, or immune checkpoint receptors. Antibodies that bind to and block or inhibit ligands can be mentioned. Exemplary checkpoint molecules that can be targeted for blockage or inhibition include CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, GAL9, LAG3, TIM3, VISTA, KIR, 2B4 (belongs to the CD2 family of molecules and is expressed on all NK, γδ, and memory CD8 ⁺ (αβ) T cells). ), CD160 (also called BY55), CGEN-15049, CHK1 and CHK2 kinases, A2aR, and various B-7 family ligands. B7 family ligands include B7-1, B7-2, B7-DC, B7-H1, B7-H2, B7-H3, B7-H4, B7-H5, B7-H6, and B7-H7. , but not limited to. Checkpoint inhibitors include those that bind to one or more of CTLA-4, PDL1, PDL2, PD1, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, and CGEN-15049; Antibodies, or antigen-binding fragments thereof, other binding proteins, biological therapeutics, or small molecules that block or inhibit the activity of. Exemplary immune checkpoint inhibitors include tremelimumab (CTLA-4 blocking antibody), anti-OX40, PD-L1 monoclonal antibody (anti-B7-Hl; MEDI4736), MK-3475 (PD-1 blocking antibody), 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 inhibitors). Checkpoint protein ligands include, but are not limited to, PD-L1, PD-L2, B7-H3, B7-H4, CD28, CD86, and TIM-3.

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 nivolumab (

Opdivo®), ipilimumab (Yervoy®), and pembrolizumab (Keytruda®). In some embodiments, the checkpoint inhibitor is nivolumab (anti-PD-1 antibody, Opdivo®, Bristol-Myers Squibb); pembrolizumab (anti-PD-1 antibody, Keytruda®, Merck); Ipilimumab (anti-CTLA-4 antibody, Yervoy®, Bristol-Myers Squibb); durvalumab (anti-PD-L1 antibody, Imfinzi®, AstraZeneca); and atezolizumab (anti-PD-L1 antibody, Tecentriq®) Trademark), Genentech).

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

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

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).

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

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 and REGN3767 and IMP321. BMS-986016 (Bristol-Myers Squibb), an anti-LAG-3 antibody, is being studied in glioblastoma and gliosarcoma (NCT02658981). REGN3767 (Regeneron) is also an anti-LAG-3 antibody and is being studied in malignancies (NCT03005782). IMP321 (Immutep S.A.) is a LAG-3-Ig fusion protein that has been studied in melanoma (NCT02676869); adenocarcinoma (NCT02614833); and metastatic breast cancer (NCT00349934).

Checkpoint inhibitors that can be used in the present invention include OX40 agonists. OX40 agonists being investigated in clinical trials include PF-04518600/PF-8600 (Pfizer), an agonist anti-OX40 antibody in metastatic kidney cancer (NCT03092856) and advanced cancers and tumors (NCT02554812; NCT05082566); GSK3174998 (Merck), an agonist anti-OX40 antibody in phase 1 cancer trials (NCT02528357); MEDI0562 (Medimune/AstraZenec), an agonist anti-OX40 antibody in advanced solid tumors (NCT02318394 and NCT02705482); a); Colorectal MEDI6469 (Medimmune/AstraZeneca), an agonist anti-OX40 antibody in patients with cancer (NCT02559024), breast cancer (NCT01862900), head and neck cancer (NCT02274155), and metastatic prostate cancer (NCT01303705); CT02737475) Examples include BMS-986178 (Bristol-Myers Squibb), which is an agonist anti-OX40 antibody.

Checkpoint inhibitors that can be used in the present invention include CD137 (also referred to as 4-1BB). CD137 agonists being investigated in clinical trials include utmilumab (PF-05082566, Pfizer ); urelumab (BMS-663513, Bristol-Myers Squibb), an agonist anti-CD137 antibody in melanoma and skin cancer (NCT02652455), and glioblastoma and gliosarcoma (NCT02658981); and metastatic or locally advanced malignancies CTX-471 (Compass Therapeutics), an agonist anti-CD137 antibody in tumors (NCT03881488).

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

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 agonistic anti-GITR antibody in malignant melanoma and other malignant solid tumors (NCT01239134 and NCT02628574); GWN323 (Novartis), an agonist anti-GITR antibody in advanced cancers (NCT02697591 and NCT03126110); INCAGN01876 (Incyte/Agenus), an agonist anti-GITR antibody in advanced cancers (NCT02697591 and NCT03126110); is an agonist anti-GITR antibody , MK-4166 (Merck), and MEDI1873 (Medimune/AstraZeneca), an agonistic hexameric GITR-ligand molecule containing a human IgG1 Fc domain, in advanced solid tumors (NCT02583165).

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

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

Checkpoint inhibitors that can be used in the present invention include CD47 inhibitors of the interaction between CD47 and signal regulatory protein alpha (SIRPa). CD47/SIRPa inhibitors being studied in clinical trials include ALX-148, an antagonist variant of (SIRPa) that binds to CD47 and prevents CD47/SIRPa-mediated signaling, in phase 1 (NCT03013218); Created by combining the N-terminal CD47-binding domain of SIRPa with the Fc domain of human IgG1, it acts by binding to human CD47 and prevents it from delivering a "don't eat" signal to macrophages; TTI-621 (SIRPa-Fc, Trillium Therapeutics), a soluble recombinant fusion protein in phase I clinical trials (NCT02890368 and NCT02663518); CC-90002 (Celgene), an anti-CD4 antibody in leukemia (NCT02641002); ); and Hu5F9-G4 (Forty Seven, Inc.) in colon and solid tumors (NCT02953782), acute myeloid leukemia (NCT02678338), and lymphoma (NCT02953509).

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

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

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

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

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

The following examples are intended to illustrate the invention and should not be construed as a limitation thereon. Unless otherwise stated, one or more tautomeric forms of the compounds of the Examples described herein below can be prepared and/or isolated in situ. All tautomeric forms of the compounds of the Examples described herein below are to be considered as disclosed. Temperature is expressed in degrees Celsius. Unless otherwise stated, all evaporations are carried out under reduced pressure, preferably between about 15 mmHg and 100 mmHg (20-133 mbar). The structure of the final products, intermediates, and starting materials is confirmed by standard analytical techniques, such as microanalysis and spectroscopic properties, such as MS, IR, NMR. The abbreviations used are conventional in the art.

The starting materials, building blocks, reagents, acids, bases, dehydrating agents, solvents, and catalysts used to synthesize the compounds of the invention are all commercially available or prepared by organic synthesis methods known to those skilled in the art. It's either possible. Additionally, the compounds of the invention can be made by organic synthesis methods known to those skilled in the art, as illustrated in the Examples below.

Example 1: Synthesis of Exemplary Compounds Certain exemplary compounds are prepared according to the following scheme.
I-1

ＬｉＡｌＨ_４（８４．２８ｍｇ、２．２２ｍｍｏｌ、２当量）のＴＨＦ（５ｍＬ）溶液に、３－（トリフルオロメチル）ビシクロ［１．１．１］ペンタン－１－カルボン酸（２００ｍｇ、１．１１ｍｍｏｌ、１当量）を０℃を加え、２５℃で２時間撹拌した。ＴＬＣ（ＰＥ／ＥｔＯＡｃ＝１／１、Ｒ_ｆ＝０．６０）は、出発物質が完全に消費され、新しいスポットが１つ形成されたことを示した。混合物を１０％のＮａＯＨ水溶液（０．５ｍＬ）でクエンチし、沈殿した固体を濾過して、濾液を濃縮し、［３－（トリフルオロメチル）－１－ビシクロ［１．１．１］ペンタニル］メタノール（１８０ｍｇ、９７５．０８μｍｏｌ、収率８７．８％、純度９０％）を無色の油として得、これをさらに精製することなく次工程で使用した。^１Ｈ ＮＭＲ（５００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ３．６７（ｓ，２Ｈ），１．９２（ｓ，６Ｈ）． Step 1: [3-(trifluoromethyl)-1-bicyclo[1.1.1]pentanyl]methanol

To a solution of LiAlH ₄ (84.28 mg, 2.22 mmol, 2 eq.) in THF (5 mL) was added 3-(trifluoromethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (200 mg, 1.11 mmol, 1 equivalent) was added at 0°C and stirred at 25°C for 2 hours. TLC (PE/EtOAc=1/1, R _f =0.60) showed complete consumption of starting material and formation of one new spot. The mixture was quenched with 10% aqueous NaOH (0.5 mL), the precipitated solid was filtered, and the filtrate was concentrated to give [3-(trifluoromethyl)-1-bicyclo[1.1.1]pentanyl] Methanol (180 mg, 975.08 μmol, 87.8% yield, 90% purity) was obtained as a colorless oil, which was used in the next step without further purification. ^1H NMR (500 MHz, _CDCl3 ) δ ppm 3.67 (s, 2H), 1.92 (s, 6H).

［３－（トリフルオロメチル）－１－ビシクロ［１．１．１］ペンタニル］メタノール（１８０ｍｇ、９７５．０８μｍｏｌ、純度９０％、１当量）及びＥｔ_３Ｎ（１９７．３３ｍｇ、１．９５ｍｍｏｌ、２７１．４４μＬ、２当量）の、ＤＣＭ（３ｍＬ）溶液に、ＭｓＣｌ（１７０ｍｇ、１．４８ｍｍｏｌ、１１４．８６μＬ、１．５２当量）を０℃で滴下して、混合物を２５℃で２時間撹拌した。ＴＬＣ（ＰＥ／ＥｔＯＡｃ＝１／１、Ｒ_ｆ＝０．６７）は、出発物質が完全に消費され、新しいスポットが１つ形成されたことを示した。混合物を飽和ＮａＨＣＯ_３水溶液（１０ｍＬ）でクエンチし、ＤＣＭで抽出した（１５ｍＬ×２）。合わせた有機相を無水Ｎａ_２ＳＯ_４で乾燥させ、濾過し、濃縮して、［３－（トリフルオロメチル）－１－ビシクロ［１．１．１］ペンタニル］メチルメタンスルホネート（２３０ｍｇ、８９４．６５μｍｏｌ、収率９１．８％、純度９５．０％）を無色ゴムとして得、これをさらに精製することなく、次工程で使用した。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ４．２５（ｓ，２Ｈ），３．０２（ｓ，３Ｈ），２．０２（ｓ，６Ｈ）． Step 2: [3-(trifluoromethyl)-1-bicyclo[1.1.1]pentanyl]methyl methanesulfonate

[3-(trifluoromethyl)-1-bicyclo[1.1.1]pentanyl]methanol (180 mg, 975.08 μmol, purity 90%, 1 eq.) and Et ₃ N (197.33 mg, 1.95 mmol, 271 To a solution of .44 μL, 2 eq.) in DCM (3 mL) was added MsCl (170 mg, 1.48 mmol, 114.86 μL, 1.52 eq.) dropwise at 0° C. and the mixture was stirred at 25° C. for 2 h. TLC (PE/EtOAc=1/1, R _f =0.67) showed complete consumption of starting material and formation of one new spot. The mixture was quenched with saturated _aqueous NaHCO (10 mL) and extracted with DCM (15 mL x 2). The combined organic phases were dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated to give [3-(trifluoromethyl)-1-bicyclo[1.1.1]pentanyl]methyl methanesulfonate (230 mg, 894. 65 μmol, yield 91.8%, purity 95.0%) was obtained as a colorless rubber, which was used in the next step without further purification. ¹ H NMR (400 MHz, _CDCl3 ) δ ppm 4.25 (s, 2H), 3.02 (s, 3H), 2.02 (s, 6H).

Ｎ－［（１Ｓ，５Ｒ）－３－アザビシクロ［３．１．０］ヘキサン－６－イル］プロパ－２－エナミド（１２０ｍｇ、３６０．６１μｍｏｌ、純度８０％、１当量、ＴＦＡ）、及び［３－（トリフルオロメチル）－１－ビシクロ［１．１．１］ペンタニル］メチルメタンスルホネート（９２．７１ｍｇ、３６０．６１μｍｏｌ、純度９５％、１当量）の、ＡＣＮ（３ｍＬ）溶液に、Ｋ_２ＣＯ_３（１４９．５２ｍｇ、１．０８ｍｍｏｌ、３当量）及びＫＩ（５．９９ｍｇ、３６．０６μｍｏｌ、０．１当量）を加えた。混合物を６０℃で１６時間撹拌した。溶媒を除去して残渣を得、これを分取ＨＰＬＣ（カラム：Ｗｅｌｃｈ Ｘｔｉｍａｔｅ Ｃ１８ １５０＊２５ｍｍ＊５ｕｍ；移動相：［水（１０ｍＭのＮＨ_４ＨＣＯ_３）－ＡＣＮ］；Ｂ％：４８％－７８％、１０分）により精製し、続いて凍結乾燥により、Ｎ－［（１Ｓ，５Ｒ）－３－［［３－（トリフルオロメチル）－１－ビシクロ［１．１．１］ペンタニル］メチル］－３－アザビシクロ［３．１．０］ヘキサン－６－イル］プロパ－２－エナミド（１８．２８ｍｇ、６０．８７μｍｏｌ、収率１６．９％、純度１００．０％）を白色固体として得た。^１Ｈ ＮＭＲ（５００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ６．２６（ｄ，Ｊ＝１６．９ Ｈｚ，１Ｈ），６．０１（ｄｄ，Ｊ＝１０．４，１６．９ Ｈｚ，１Ｈ），５．６１（ｄ，Ｊ＝１０．２Ｈｚ，１Ｈ），５．５４（ｂｒ ｓ，１Ｈ），３．１７（ｄ，Ｊ＝８．９ Ｈｚ，２Ｈ），３．０２（ｄ，Ｊ＝２．０ Ｈｚ，１Ｈ），２．５１（ｓ，２Ｈ），２．３７（ｄ，Ｊ＝ ８．４ Ｈｚ，２Ｈ），１．８６（ｓ，６Ｈ），１．６１（ｓ，２Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ３０１．２［Ｍ＋Ｈ］^＋． Step 3: N-[(1S,5R)-3-[[3-(trifluoromethyl)-1-bicyclo[1.1.1]pentanyl]methyl]-3-azabicyclo[3.1.0]hexane -6-yl]prop-2-enamide

N-[(1S,5R)-3-azabicyclo[3.1.0]hexan-6-yl]prop-2-enamide (120 mg, 360.61 μmol, 80% purity, 1 eq., TFA), and [3 K ₂ CO ₃ (149.52 mg, 1.08 mmol, 3 eq.) and KI (5.99 mg, 36.06 μmol, 0.1 eq.) were added. The mixture was stirred at 60°C for 16 hours. The solvent was removed to obtain a residue, which was subjected to preparative HPLC (column: Welch Xtimate C18 150*25mm*5um; mobile phase: [water (10mM NH ₄ HCO ₃ )-ACN]; B%: 48%-78 %, 10 min) followed by lyophilization to give N-[(1S,5R)-3-[[3-(trifluoromethyl)-1-bicyclo[1.1.1]pentanyl]methyl] -3-Azabicyclo[3.1.0]hexan-6-yl]prop-2-enamide (18.28 mg, 60.87 μmol, yield 16.9%, purity 100.0%) was obtained as a white solid. . ^1H NMR (500 MHz, _CDCl3 ) δ ppm 6.26 (d, J=16.9 Hz, 1H), 6.01 (dd, J=10.4, 16.9 Hz, 1H), 5. 61 (d, J = 10.2 Hz, 1H), 5.54 (br s, 1H), 3.17 (d, J = 8.9 Hz, 2H), 3.02 (d, J = 2.0 Hz, 1H), 2.51 (s, 2H), 2.37 (d, J = 8.4 Hz, 2H), 1.86 (s, 6H), 1.61 (s, 2H); ES- LCMS m/z 301.2 [M+H] ⁺ .

工程１：［４－（トリフルオロメチル）－１－ビシクロ［２．２．２］オクタニル］メタノール
４－（トリフルオロメチル）ビシクロ［２．２．２］オクタン－１－カルボン酸（３００ｍｇ、１．３５ｍｍｏｌ、１当量）のＴＨＦ（５ｍＬ）溶液に、ＬｉＡｌＨ_４（１０２．４９ｍｇ、２．７０ｍｍｏｌ、２当量）を２５℃で加えた。混合物を２５℃で１時間撹拌した。ＴＬＣ（ＰＥ／ＥｔＯＡｃ＝３／１、Ｒ_ｆ＝０．５９）は、出発物質が完全に消費され、新しいスポットが１つ形成されたことを示した。混合物を１０％のＮａＯＨ水溶液（０．５ｍＬ）でクエンチし、沈殿した固体を濾過して、濾液を濃縮し、［４－（トリフルオロメチル）－１－ビシクロ［２．２．２］オクタニル］メタノール（２８０ｍｇ、１．２８μｍｏｌ、収率９４．６％、純度９５．０％）を無色の油として得、これをさらに精製することなく次工程で使用した。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ３．３０（ｓ，２Ｈ），１．７６－１．６５（ｍ，６Ｈ），１．５１－１．４１（ｍ，６Ｈ）． I-8

Step 1: [4-(trifluoromethyl)-1-bicyclo[2.2.2]octanyl]methanol
LiAlH ₄ (102.49 mg, 2.70 mmol, 2 equivalents) was added at 25°C. The mixture was stirred at 25°C for 1 hour. TLC (PE/EtOAc=3/1, R _f =0.59) showed complete consumption of starting material and formation of one new spot. The mixture was quenched with 10% aqueous NaOH (0.5 mL), the precipitated solid was filtered, and the filtrate was concentrated to give [4-(trifluoromethyl)-1-bicyclo[2.2.2]octanyl] Methanol (280 mg, 1.28 μmol, 94.6% yield, 95.0% purity) was obtained as a colorless oil, which was used in the next step without further purification. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 3.30 (s, 2H), 1.76-1.65 (m, 6H), 1.51-1.41 (m, 6H).

［４－（トリフルオロメチル）－１－ビシクロ［２．２．２］オクタニル］メタノール（５０ｍｇ、２２８．１２μｍｏｌ、純度９５％、１当量）のＤＣＭ（３ｍＬ）溶液に、ＰＣＣ（９８．３５ｍｇ、４５６．２５μｍｏｌ、２当量）を加えた。混合物を２５℃で１時間撹拌した。ＴＬＣ（ＰＥ／ＥｔＯＡｃ＝３／１、Ｒ_ｆ＝０．７５）は、出発物質が完全に消費され、新しいスポットが１つ形成されたことを示した。溶媒を除去して残渣を得、これをフラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝２／１から、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝３／１、Ｒ_ｆ＝０．７５）により精製し、４－（トリフルオロメチル）ビシクロ［２．２．２］オクタン－１－カルバルデヒド（５０ｍｇ、２２３．０８μｍｏｌ、収率９７．８％、純度９２．０％）を黄色油として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ９．４７（ｓ，１Ｈ），１．７６－１．６９（ｍ，１２Ｈ）． Step 2: 4-(trifluoromethyl)bicyclo[2.2.2]octane-1-carbaldehyde

PCC (98.35 mg, 456.25 μmol, 2 equivalents) were added. The mixture was stirred at 25°C for 1 hour. TLC (PE/EtOAc=3/1, R _f =0.75) showed complete consumption of starting material and formation of one new spot. Removal of the solvent gave a residue, which was purified by flash silica gel chromatography (from PE/EtOAc=2/1, TLC:PE/EtOAc=3/1, R _f =0.75) to 4-(tri (fluoromethyl)bicyclo[2.2.2]octane-1-carbaldehyde (50 mg, 223.08 μmol, yield 97.8%, purity 92.0%) was obtained as a yellow oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.47 (s, 1H), 1.76-1.69 (m, 12H).

Ｎ－［（１Ｓ，５Ｒ）－３－アザビシクロ［３．１．０］ヘキサン－６－イル］プロパ－２－エナミド（１２０ｍｇ、３６０．６１μｍｏｌ、純度８０％、１当量、ＴＦＡ）及びＴＥＡ（３６．４９ｍｇ、３６０．６１μｍｏｌ、５０．１９μＬ、１当量）の、ＭｅＯＨ（１０ｍＬ）溶液に、４－（トリフルオロメチル）ビシクロ［２．２．２］オクタン－１－カルバルデヒド（４７．４３ｍｇ、２１１．６２μｍｏｌ、純度９２％）を加えた。混合物を２５℃で２時間撹拌した。ＮａＢＨ_３ＣＮ（６７．９８ｍｇ、１．０８ｍｍｏｌ、３当量）を加え、混合物を２５℃で１６時間撹拌した。溶媒を除去して残渣を得、これを分取ＨＰＬＣ（カラム：Ａｇｅｌａ ＤｕｒａＳｈｅｌｌ Ｃ１８ １５０＊２５ｍｍ＊５ｕｍ；移動相：［水（０．０５％のＮＨ_３Ｈ_２Ｏ＋１０ｍＭのＮＨ_４ＨＣＯ_３）－ＡＣＮ］；Ｂ％：５１％－８１％、１０分）により精製し、続いて凍結乾燥して、Ｎ－［（１Ｒ，５Ｓ）－３－［［４－（トリフルオロメチル）－１－ビシクロ［２．２．２］オクタニル］メチル］－３－アザビシクロ［３．１．０］ヘキサン－６－イル］プロパ－２－エナミド（２４．６４ｍｇ、７１．９６μｍｏｌ、収率２０．０％、純度１００．０％）を白色固体として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ６．３１－６．２２（ｍ，１Ｈ），６．０１（ｄｄ，Ｊ＝１０．３，１７．１ Ｈｚ，１Ｈ），５．６１（ｄｄ，Ｊ＝１．１，１０．１ Ｈｚ，１Ｈ），５．４９（ｂｒ ｓ，１Ｈ），３．１３（ｄ，Ｊ＝８．８ Ｈｚ，２Ｈ），３．０２（ｄ，Ｊ＝１．７Ｈｚ，１Ｈ），２．４８（ｄ，Ｊ＝８．３ Ｈｚ，２Ｈ），２．１３（ｓ，２Ｈ），１．６８－１．５９（ｍ，６Ｈ），１．４９（ｓ，２Ｈ），１．４３－１．３２（ｍ，６Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ３４３．２［Ｍ＋Ｈ］^＋． Step 3: N-[(1R,5S)-3-[[4-(trifluoromethyl)-1-bicyclo[2.2.2]octanyl]methyl]-3-azabicyclo[3.1.0]hexane -6-yl]prop-2-enamide

N-[(1S,5R)-3-azabicyclo[3.1.0]hexan-6-yl]prop-2-enamide (120 mg, 360.61 μmol, 80% purity, 1 eq., TFA) and TEA (36 4-(trifluoromethyl)bicyclo[2.2.2]octane-1-carbaldehyde (47.43 mg, 211 .62 μmol, purity 92%) was added. The mixture was stirred at 25°C for 2 hours. NaBH ₃ CN (67.98 mg, 1.08 mmol, 3 eq.) was added and the mixture was stirred at 25° C. for 16 hours. The solvent was removed to obtain a residue, which was subjected to preparative HPLC (column: Agela DuraShell C18 150*25mm*5um; mobile phase: [water (0.05% NH ₃ H ₂ O + 10 mM NH ₄ HCO ₃ )-ACN ]; B%: 51%-81%, 10 min) followed by lyophilization to give N-[(1R,5S)-3-[[4-(trifluoromethyl)-1-bicyclo[ 2.2.2]octanyl]methyl]-3-azabicyclo[3.1.0]hexan-6-yl]prop-2-enamide (24.64 mg, 71.96 μmol, yield 20.0%, purity 100 .0%) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 6.31-6.22 (m, 1H), 6.01 (dd, J=10.3, 17.1 Hz, 1H), 5.61 (dd , J=1.1, 10.1 Hz, 1H), 5.49 (br s, 1H), 3.13 (d, J=8.8 Hz, 2H), 3.02 (d, J=1 .7Hz, 1H), 2.48 (d, J=8.3 Hz, 2H), 2.13 (s, 2H), 1.68-1.59 (m, 6H), 1.49 (s, 2H), 1.43-1.32 (m, 6H); ES-LCMS m/z 343.2 [M+H] ⁺ .

２－ブロモ－４－ニトロ－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］アニリン（１．１０ｇ、２．６７ｍｍｏｌ、純度９１．１％、１当量）のＤＣＭ（１０ｍＬ）溶液に、（Ｂｏｃ）_２Ｏ（１．７５ｇ、８．００ｍｍｏｌ、１．８４ｍＬ、３当量）及びＤＭＡＰ（３２５．６６ｍｇ、２．６７ｍｍｏｌ、１当量）を加えた。混合物を２５℃で１２時間撹拌した。反応混合物を濃縮して残渣を得、これをフラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１／０から１０／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝１０／１、Ｒ_ｆ＝０．５１）により精製して、ｔｅｒｔ－ブチル＝Ｎ－（２－ブロモ－４－ニトロ－フェニル）－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］カルバマート（１．１５ｇ、２．４０ｍｍｏｌ、収率８９．９％、純度９８．７％）を白色固体として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３） δ ｐｐｍ ８．４９（ｂｒ ｓ，１Ｈ），８．０６（ｄ，Ｊ＝７．８ Ｈｚ， １Ｈ），７．５７（ｄ，Ｊ＝７．４ Ｈｚ，２Ｈ），７．３６（ｄ，Ｊ＝８．２ Ｈｚ，２Ｈ），５．２０（ｄ，Ｊ＝１４．１ Ｈｚ，１Ｈ），４．３９（ｄ，Ｊ＝１５．７ Ｈｚ，１Ｈ），１．５７－１．４０（ｍ，９Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ３７５．３［Ｍ－Ｂｏｃ＋Ｈ］^＋． I-9
Step 1: tert-butyl=(2-bromo-4-nitrophenyl)(4-(trifluoromethyl)benzyl)carbamate

A solution of 2-bromo-4-nitro-N-[[4-(trifluoromethyl)phenyl]methyl]aniline (1.10 g, 2.67 mmol, 91.1% purity, 1 eq.) in DCM (10 mL) (Boc) ₂ O (1.75 g, 8.00 mmol, 1.84 mL, 3 eq.) and DMAP (325.66 mg, 2.67 mmol, 1 eq.) were added. The mixture was stirred at 25°C for 12 hours. The reaction mixture was concentrated to give a residue, which was purified by flash silica gel chromatography (PE/EtOAc=1/0 to 10/1, TLC:PE/EtOAc=10/1, R _f =0.51). , tert-butyl N-(2-bromo-4-nitro-phenyl)-N-[[4-(trifluoromethyl)phenyl]methyl]carbamate (1.15 g, 2.40 mmol, yield 89.9% , purity 98.7%) was obtained as a white solid. ^1H NMR (400 MHz, _CDCl3 ) δ ppm 8.49 (br s, 1H), 8.06 (d, J = 7.8 Hz, 1H), 7.57 (d, J = 7.4 Hz , 2H), 7.36 (d, J = 8.2 Hz, 2H), 5.20 (d, J = 14.1 Hz, 1H), 4.39 (d, J = 15.7 Hz, 1H) ), 1.57-1.40 (m, 9H); ES-LCMS m/z 375.3 [M-Boc+H] ⁺ .

ｔｅｒｔ－ブチル＝Ｎ－（２－ブロモ－４－ニトロ－フェニル）－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］カルバマート（１．１１ｇ、２．３１ｍｍｏｌ、９８．７％、１当量）の、ＥｔＯＨ（５ｍＬ）及びＨ_２Ｏ（５ｍＬ）溶液に、Ｆｅ（６４６．２８ｍｇ、１１．５７ｍｍｏｌ、５当量）及びＮＨ_４Ｃｌ（１．２４ｇ、２３．１５ｍｍｏｌ、１０当量）を加えた。混合物を８０℃で１時間撹拌した。水（４０ｍＬ）を添加することにより反応混合物をクエンチし、ＥｔＯＡｃで抽出した（３０ｍＬ×３）。合わせた有機層をブライン（１０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して、ｔｅｒｔ－ブチル＝Ｎ－（４－アミノ－２－ブロモ－フェニル）－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］カルバマート（１．０１ｇ、２．０６ｍｍｏｌ、収率８９．１％、純度９１．０％）を黄色固体として得、これをさらに精製することなく、次工程で使用した。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ７．５４（ｄ，Ｊ＝７．８ Ｈｚ，２Ｈ），７．３９（ｓ，２Ｈ），６．９５－６．８８（ｍ，１Ｈ），６．５８（ｄ，Ｊ＝８．６ Ｈｚ，１Ｈ），６．５０－６．３９（ｍ，１Ｈ），５．２０（ｄ，Ｊ＝１４．９ Ｈｚ，１Ｈ），４．２７－４．２０（ｍ，１Ｈ），３．７２（ｓ，２Ｈ），１．３８（ｓ，９Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ３９１．０［Ｍ－Ｂｏｃ＋Ｈ］^＋． Step 2: tert-butyl(4-amino-2-bromophenyl)(4-(trifluoromethyl)benzyl)carbamate

tert-Butyl N-(2-bromo-4-nitro-phenyl)-N-[[4-(trifluoromethyl)phenyl]methyl]carbamate (1.11 g, 2.31 mmol, 98.7%, 1 eq. ) in EtOH (5 mL) and H ₂ O (5 mL) were added Fe (646.28 mg, 11.57 mmol, 5 eq.) and NH ₄ Cl (1.24 g, 23.15 mmol, 10 eq.). The mixture was stirred at 80°C for 1 hour. The reaction mixture was quenched by adding water (40 mL) and extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine (10 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give tert-butyl=N-(4-amino-2-bromo-phenyl)-N-[[ 4-(Trifluoromethyl)phenyl]methyl]carbamate (1.01 g, 2.06 mmol, 89.1% yield, 91.0% purity) was obtained as a yellow solid, which was carried into the next step without further purification. It was used in ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.54 (d, J = 7.8 Hz, 2H), 7.39 (s, 2H), 6.95-6.88 (m, 1H), 6.58 (d, J = 8.6 Hz, 1H), 6.50-6.39 (m, 1H), 5.20 (d, J = 14.9 Hz, 1H), 4.27-4 .20 (m, 1H), 3.72 (s, 2H), 1.38 (s, 9H); ES-LCMS m/z 391.0 [M-Boc+H] ⁺ .

ｔｅｒｔ－ブチル＝Ｎ－（４－アミノ－２－ブロモ－フェニル）－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］カルバマート（２５０ｍｇ、５３９．００μｍｏｌ、純度９６％、１当量）のＤＣＭ（１０ｍＬ）溶液に、塩化アクリロイル（７３．１８ｍｇ、８０８．４９μｍｏｌ、６５．９２μＬ、１．５当量）、及びＤＩＥＡ（１３９．３２ｍｇ、１．０８ｍｍｏｌ、１８７．７７μＬ、２当量）を加えた。混合物を２５℃で１時間撹拌した。水（５０ｍＬ）を添加することにより反応混合物をクエンチし、ＥｔＯＡｃで抽出した（５０ｍＬ×３）。合わせた有機層をブライン（１０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、フラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１／０から３／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝３／１、Ｒ_ｆ＝０．４３）により精製し、ｔｅｒｔ－ブチルＮ－［２－ブロモ－４－（プロパ－２－エノイルアミド）フェニル］－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］カルバマート（２４３ｍｇ、４８６．６６μｍｏｌ、収率９０．２％、純度１００％）を無色の油として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ７．６０－７．５３（ｍ，２Ｈ），７．４０－７．２９（ｍ，３Ｈ），７．２６－７．１８（ｍ，１Ｈ），６．９２－６．７５（ｍ，１Ｈ），６．４６（ｄ，Ｊ＝１６．８ Ｈｚ，１Ｈ），６．２１（ｄｄ，Ｊ＝１０．６，１６．８ Ｈｚ，１Ｈ），５．８４－５．７４（ｍ，１Ｈ），５．２１（ｄ，Ｊ＝１５．３ Ｈｚ，１Ｈ），４．３０（ｓ，１Ｈ），４．２６（ｓ，１Ｈ），４．１３（ｑ，Ｊ＝７．２ Ｈｚ，１Ｈ），２．０６（ｓ，１Ｈ），１．６３－１．５２（ｍ，９Ｈ），１．３３－１．２５（ｍ，２Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ５２１．１［Ｍ＋Ｈ］^＋． Step 3: tert-butyl(4-acrylamido-2-bromophenyl)(4-(trofluorophenyl)benzyl)carbamate

tert-Butyl N-(4-amino-2-bromo-phenyl)-N-[[4-(trifluoromethyl)phenyl]methyl]carbamate (250 mg, 539.00 μmol, 96% purity, 1 eq.) in DCM (10 mL) solution was added acryloyl chloride (73.18 mg, 808.49 μmol, 65.92 μL, 1.5 eq.) and DIEA (139.32 mg, 1.08 mmol, 187.77 μL, 2 eq.). The mixture was stirred at 25°C for 1 hour. The reaction mixture was quenched by adding water (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (10 mL), dried over _Na2SO4 , filtered and concentrated _in vacuo to give a residue, which was purified by flash silica gel chromatography (PE/EtOAc=1/0 to 3/1, TLC: tert-butyl N-[2-bromo-4-(prop-2-enoylamido)phenyl] _-N -[[4-(trifluoro Methyl)phenyl]methyl]carbamate (243 mg, 486.66 μmol, 90.2% yield, 100% purity) was obtained as a colorless oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.60-7.53 (m, 2H), 7.40-7.29 (m, 3H), 7.26-7.18 (m, 1H) , 6.92-6.75 (m, 1H), 6.46 (d, J = 16.8 Hz, 1H), 6.21 (dd, J = 10.6, 16.8 Hz, 1H), 5.84-5.74 (m, 1H), 5.21 (d, J=15.3 Hz, 1H), 4.30 (s, 1H), 4.26 (s, 1H), 4.13 (q, J=7.2 Hz, 1H), 2.06 (s, 1H), 1.63-1.52 (m, 9H), 1.33-1.25 (m, 2H); ES- LCMS m/z 521.1 [M+H] ⁺ .

ｔｅｒｔ－ブチルＮ－［２－ブロモ－４－（プロパ－２－エノイルアミド）フェニル］－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］カルバマート（２００ｍｇ、４００．５４μｍｏｌ、１００％、１当量）及びトリブチル－（１－メチルイミダゾール－４－イル）スタンナン（２３４．７３ｍｇ、６００．８２μｍｏｌ，、９５％、１．５当量）の、ＤＭＦ（７ｍＬ）溶液に、Ｐｄ（ｄｐｐｆ）Ｃｌ_２（２９．３１ｍｇ、４０．０５μｍｏｌ、０．１当量）を加えた。混合物を、Ｎ_２雰囲気下、１３０℃で１時間撹拌した。水（５０ｍＬ）を添加することにより反応混合物をクエンチし、ＥｔＯＡｃで抽出した（５０ｍＬ×３）。合わせた有機層をブライン（２０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、フラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１／０から４／５、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝３／１、Ｒ_ｆ＝０．２０）により精製し、ｔｅｒｔ－ブチル＝Ｎ－［２－（１－メチルイミダゾール－４－イル）－４－（プロパ－２－エノイルアミド）フェニル］－Ｎ－［［４－（トリフルオロフェニル）フェニル］メチル］カルバマート（１００ｍｇ、１９９．８０μｍｏｌ、収率４９．８％、純度１００．０％）を無色の油として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ７．８６（ｓ，１Ｈ），７．５５－７．４３（ｍ，４Ｈ），７．３４（ｄ，Ｊ＝７．８ Ｈｚ，３Ｈ），６．８８（ｂｒ ｓ，１Ｈ），６．７３（ｄ，Ｊ＝７．０ Ｈｚ，１Ｈ），６．４５－６．３５（ｍ，１Ｈ），６．２５－６．１４（ｍ，１Ｈ），５．７６（ｄ，Ｊ＝１０．６ Ｈｚ，１Ｈ），５．１８（ｄ，Ｊ＝１４．５ Ｈｚ，１Ｈ），４．１８－４．０７（ｍ，１Ｈ），４．１１（ｄ，Ｊ＝１３．７ Ｈｚ，１Ｈ），３．６７（ｓ，３Ｈ），１．２６（ｓ，９Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ５０１．２［Ｍ＋Ｈ］^＋． Step 4: tert-butyl(4-acrylamido-2-(1-methyl-1H-imidazol-4-yl)phenyl)(4-(trifluoromethyl)benzyl)carbamate

tert-Butyl N-[2-bromo-4-(prop-2-enoylamido)phenyl]-N-[[4-(trifluoromethyl)phenyl]methyl]carbamate (200 mg, 400.54 μmol, 100%, 1 eq. Pd(dppf)Cl ₂ (29 .31 mg, 40.05 μmol, 0.1 equivalent) was added. The mixture was stirred at 130 °C for 1 h under _N2 atmosphere. The reaction mixture was quenched by adding water (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was subjected to flash silica gel chromatography (PE/EtOAc = 1/0 to 4/5, TLC: tert-butyl N-[2-(1-methylimidazol-4-yl)-4-(prop _- 2-enoylamido)phenyl] -N-[[4-(trifluorophenyl)phenyl]methyl]carbamate (100 mg, 199.80 μmol, yield 49.8%, purity 100.0%) was obtained as a colorless oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.86 (s, 1H), 7.55-7.43 (m, 4H), 7.34 (d, J = 7.8 Hz, 3H), 6.88 (br s, 1H), 6.73 (d, J=7.0 Hz, 1H), 6.45-6.35 (m, 1H), 6.25-6.14 (m, 1H ), 5.76 (d, J = 10.6 Hz, 1H), 5.18 (d, J = 14.5 Hz, 1H), 4.18-4.07 (m, 1H), 4.11 (d, J=13.7 Hz, 1H), 3.67 (s, 3H), 1.26 (s, 9H); ES-LCMS m/z 501.2 [M+H] ⁺ .

ｔｅｒｔ－ブチルＮ－［２－（１－メチルイミダゾール－４－イル）－４－（プロパ－２－エノイルアミド）フェニル］－Ｎ－［［４－（トリフルオロフェニル）フェニル］メチル］カルバマート（１００ｍｇ、１９９．８０μｍｏｌ、１００％、１当量）のＤＣＭ（３ｍＬ）溶液に、ＴＦＡ（３．０８ｇ、２７．０１ｍｍｏｌ、２．００ｍＬ、１３５．２０当量）を加えた。混合物を２５℃で１時間撹拌した。水（３０ｍＬ）を添加することにより反応混合物をクエンチし、ＥｔＯＡｃで抽出した（３０ｍＬ×３）。合わせた有機層をブライン（１０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、これを分取ＨＰＬＣ（カラム：Ａｇｅｌａ ＤｕｒａＳｈｅｌｌ Ｃ１８ １５０＊２５ｍｍ＊５μｍ；移動相：［水（０．０５％のＮＨ_３Ｈ_２Ｏ＋１０ｍＭのＮＨ_４ＨＣＯ_３）－ＡＣＮ］；Ｂ％：４０％－７０％、１０分）により精製し、次いで凍結乾燥して、Ｎ－［３－（１－メチルイミダゾール－４－イル）－４－［［４－（トリフルオロメチル）フェニル］メチルアミノ］フェニル］プロパ－２－エナミド（１０．４８ｍｇ、２６．１７μｍｏｌ、収率１３．１％、純度１００．０％）を黄色固体として得た。^１Ｈ ＮＭＲ （４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．３５（ｂｒ ｓ，１Ｈ），７．９２（ｓ，１Ｈ），７．５７－７．４５（ｍ，５Ｈ），７．２４（ｂｒ ｓ，１Ｈ），７．０８－６．９４（ｍ，２Ｈ），６．４７－６．３６（ｍ，２Ｈ），６．２６－６．１４（ｍ，１Ｈ），５．７１（ｄ，Ｊ＝１１．３ Ｈｚ，１Ｈ），４．５３（ｓ，２Ｈ），３．７４（ｓ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４０１．２［Ｍ＋Ｈ］^＋． Step 5: N-(3-(1-methyl-1H-imidazol-4-yl)-4-((4-(trifluoromethyl)benzyl)amino)phenyl)acrylamide

tert-Butyl N-[2-(1-methylimidazol-4-yl)-4-(prop-2-enoylamido)phenyl]-N-[[4-(trifluorophenyl)phenyl]methyl]carbamate (100 mg, To a solution of 199.80 μmol, 100%, 1 eq.) in DCM (3 mL) was added TFA (3.08 g, 27.01 mmol, 2.00 mL, 135.20 eq.). The mixture was stirred at 25°C for 1 hour. The reaction mixture was quenched by adding water (30 mL) and extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine (10 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Agela DuraShell C18 150*25 mm*5 μm; mobile phase : [Water (0.05% NH ₃ H ₂ O + 10 mM NH ₄ HCO ₃ )-ACN]; B%: 40%-70%, 10 min) and then lyophilized to give N-[3 -(1-Methylimidazol-4-yl)-4-[[4-(trifluoromethyl)phenyl]methylamino]phenyl]prop-2-enamide (10.48 mg, 26.17 μmol, yield 13.1%) , purity 100.0%) was obtained as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.35 (br s, 1H), 7.92 (s, 1H), 7.57-7.45 (m, 5H), 7.24 (br s , 1H), 7.08-6.94 (m, 2H), 6.47-6.36 (m, 2H), 6.26-6.14 (m, 1H), 5.71 (d, J =11.3 Hz, 1H), 4.53 (s, 2H), 3.74 (s, 3H); ES-LCMS m/z 401.2 [M+H] ⁺ .

工程１：Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－４－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］－３－ビニル－ベンゼンスルホンアミド

３－ブロモ－Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－４－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］ベンゼンスルホンアミド（６００ｍｇ、１．１３ｍｍｏｌ、１当量）及び４，４，５，５－テトラメチル－２－ビニル－１，３，２－ジオキサボロラン（３４８．４８ｍｇ、２．２６ｍｍｏｌ、３８３．７９μＬ、２当量）の、１，４－ジオキサン（１０ｍＬ）及びＨ_２Ｏ（２ｍＬ）の溶液に、Ｃｓ_２ＣＯ_３（７３７．２１ｍｇ、２．２６ｍｍｏｌ、２当量）及びＰｄ（ｄｐｐｆ）Ｃｌ_２（８２．７８ｍｇ、１１３．１３μｍｏｌ、０．１当量）を加えた。混合物を、Ｎ_２雰囲気下、１００℃で１２時間撹拌した。水（５０ｍＬ）を添加することにより反応混合物をクエンチし、ＥｔＯＡｃで抽出した（５０ｍＬ×３）。合わせた有機層をブライン（３０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、これをフラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１／０から２／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝１／１、Ｒ_ｆ＝０．５９）により精製し、Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－４－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］－３－ビニル－ベンゼンスルホンアミド（５００ｍｇ、１．０４ｍｍｏｌ、収率９１．６％、純度９８．９％）を白色固体として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．５３（ｓ，１Ｈ），７．９８－７．９０（ｍ，２Ｈ），７．７６（ｄｔ，Ｊ＝２．２，８．９ Ｈｚ，２Ｈ），７．２５（ｄ，Ｊ＝８．６ Ｈｚ，２Ｈ），６．９３－６．８２（ｍ，５Ｈ），５．８３（ｄ，Ｊ＝１７．６ Ｈｚ，１Ｈ），５．５８（ｄ，Ｊ＝１１．０ Ｈｚ，１Ｈ），４．１３（ｓ，２Ｈ），３．８１（ｓ，３Ｈ），２．６３（ｓ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４７８．６［Ｍ＋Ｈ］^＋． I-10 and I-11 (isomers of I-25)

Step 1: N-[(4-methoxyphenyl)methyl]-N-methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]-3-vinyl-benzenesulfonamide

3-bromo-N-[(4-methoxyphenyl)methyl]-N-methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]benzenesulfonamide (600 mg, 1.13 mmol, 1 eq. ) and 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (348.48 mg, 2.26 mmol, 383.79 μL, 2 eq.) in 1,4-dioxane (10 mL). and _H2O (2 mL) were added _Cs2CO3 ( _737.21 mg, 2.26 mmol, 2 eq.) and Pd(dppf) _Cl2 (82.78 mg, 113.13 μmol, 0.1 eq.). Ta. The mixture was stirred at 100 °C for 12 h under _N2 atmosphere. The reaction mixture was quenched by adding water (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give a residue, which was subjected to flash silica gel chromatography (PE/EtOAc = 1/0 to 2/1; TLC: PE/EtOAc=1/1, R _f =0.59) to give N-[(4-methoxyphenyl)methyl]-N-methyl-4-[[5-(trifluoromethyl)-2 -pyridyl]amino]-3-vinyl-benzenesulfonamide (500 mg, 1.04 mmol, yield 91.6%, purity 98.9%) was obtained as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.53 (s, 1H), 7.98-7.90 (m, 2H), 7.76 (dt, J=2.2, 8.9 Hz , 2H), 7.25 (d, J = 8.6 Hz, 2H), 6.93-6.82 (m, 5H), 5.83 (d, J = 17.6 Hz, 1H), 5 .58 (d, J=11.0 Hz, 1H), 4.13 (s, 2H), 3.81 (s, 3H), 2.63 (s, 3H); ES-LCMS m/z 478. 6 [M+H] ⁺ .

Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－４－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］－３－ビニル－ベンゼンスルホンアミド（５００ｍｇ、１．０４ｍｍｏｌ、純度９８．９％、１当量）のＤＣＭ（２ｍＬ）溶液に、ＴＦＡ（１．５２ｇ、１３．３７ｍｍｏｌ、９８９．７０μＬ、１２．９０当量）を加えた。混合物を２５℃で２時間撹拌した。水（５０ｍＬ）を添加することにより反応混合物をクエンチし、ＥｔＯＡｃで抽出した（３０ｍＬ×３）。合わせた有機層をブライン（２０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、フラッシュシリカゲルクロマトグラフィー（ｆＰＥ／ＥｔＯＡｃ＝１／０から２／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝２／１、Ｒ_ｆ＝０．４９）により精製し、Ｎ－メチル－４－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］－３－ビニル－ベンゼンスルホンアミド（３００ｍｇ、８０５．０９μｍｏｌ、収率７７．７％、純度９５．９％）を無色の油として得た。^１Ｈ ＮＭＲ （４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．５０（ｓ，１Ｈ），７．９７（ｄ，Ｊ＝２．０ Ｈｚ，１Ｈ），７．８９（ｄ，Ｊ＝８．６ Ｈｚ，１Ｈ），７．７７（ｄｔ，Ｊ＝２．１，８．５ Ｈｚ，２Ｈ），６．９４（ｓ，１Ｈ），６．９０－６．８０（ｍ，２Ｈ），５．８３（ｄ，Ｊ＝１７．４ Ｈｚ，１Ｈ），５．５６（ｄ，Ｊ＝１１．２ Ｈｚ，１Ｈ），４．５１（ｑ，Ｊ＝５．１ Ｈｚ，１Ｈ），２．７２（ｄ，Ｊ＝５．４ Ｈｚ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ３５８．１ ［Ｍ＋Ｈ］^＋． Step 2: N-methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]-3-vinyl-benzenesulfonamide

N-[(4-methoxyphenyl)methyl]-N-methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]-3-vinyl-benzenesulfonamide (500 mg, 1.04 mmol, purity To a solution of 98.9% (1 eq.) in DCM (2 mL) was added TFA (1.52 g, 13.37 mmol, 989.70 μL, 12.90 eq.). The mixture was stirred at 25°C for 2 hours. The reaction mixture was quenched by adding water (50 mL) and extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine (20 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give a residue, which was subjected to flash silica gel chromatography (fPE/EtOAc = 1/0 to 2/1, TLC: PE/EtOAc=2/1, R _f =0.49) to give N-methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]-3-vinyl-benzenesulfonamide ( 300 mg, 805.09 μmol, yield 77.7%, purity 95.9%) was obtained as a colorless oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.50 (s, 1H), 7.97 (d, J = 2.0 Hz, 1H), 7.89 (d, J = 8.6 Hz, 1H), 7.77 (dt, J = 2.1, 8.5 Hz, 2H), 6.94 (s, 1H), 6.90-6.80 (m, 2H), 5.83 (d , J=17.4 Hz, 1H), 5.56 (d, J=11.2 Hz, 1H), 4.51 (q, J=5.1 Hz, 1H), 2.72 (d, J =5.4 Hz, 3H); ES-LCMS m/z 358.1 [M+H] ⁺ .

Ｎ－メチル－４－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］－３－ビニル－ベンゼンスルホンアミド（２７０ｍｇ、７２４．５８μｍｏｌ、９５．９％、１当量）、及び、ジブロモメタノンオキシム（２９３．９４ｍｇ、１．４５ｍｍｏｌ、２当量）の、ＥｔＯＡｃ（１０ｍＬ）の撹拌溶液に、ＮａＨＣＯ_３（６０８．７０ｍｇ、７．２５ｍｍｏｌ、２８１．８０μＬ、１０当量）を加えた。反応混合物を２５℃で２時間撹拌した。水（５０ｍＬ）を添加することにより反応混合物をクエンチし、ＥｔＯＡｃで抽出した（５０ｍＬ×３）。合わせた有機層をブライン（５０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、これを分取ＨＰＬＣ（カラム：Ａｇｅｌａ ＤｕｒａＳｈｅｌｌ Ｃ１８ １５０＊２５ｍｍ＊５μｍ；移動相：［水（０．０５％のＮＨ_３・Ｈ_２Ｏ＋１０ｍＭのＮＨ_４ＨＣＯ_３）－ＡＣＮ］；Ｂ％：４０％－７０％、１０分）により精製し、次いで凍結乾燥して混合物を得、これをキラルＳＦＣカラム（ＤＡＩＣＥＬ ＣＨＩＲＡＬＰＡＫ ＩＧ （２５０ｍｍ＊３０ｍｍ、１０μｍ）；移動相：［０．１％のＮＨ_３・Ｈ_２ＯのＭＥＯＨ］；Ｂ％：４０％－４０％）により分離して、ピーク１及びピーク２を得た。ピーク１を減圧濃縮して残渣を得、これをＭｅＣＮ（１０ｍＬ）及びＨ_２Ｏ（２０ｍＬ）に溶解させ、凍結乾燥し、３－［（５Ｓ）－３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル］－Ｎ－メチル－４－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］ベンゼンスルホンアミド（４４．５１ｍｇ、９２．８７μｍｏｌ、収率１２．８％、純度１００．０％、ＳＦＣ：Ｒ_ｔ＝１．６５８、ｅｅ＝１００％、［α］^３１．４ _Ｄ＝－４４．４（ＣＨ_３ＯＨ、ｃ＝０．０５４ｇ／１００ｍＬ））を黄色固体として得た。^１Ｈ ＮＭＲ （４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．５１（ｓ，１Ｈ），８．１９（ｄ，Ｊ＝８．６ Ｈｚ，１Ｈ），７．８８（ｄｄ，Ｊ＝２．１，８．７ Ｈｚ，１Ｈ），７．８４－７．７６（ｍ，２Ｈ），７．４０（ｓ，１Ｈ），６．８１（ｄ，Ｊ＝８．８ Ｈｚ，１Ｈ），５．８１（ｔ，Ｊ＝１１．１ Ｈｚ，１Ｈ），４．４１（ｓ，１Ｈ），３．６２－３．５２（ｍ，１Ｈ），３．４９－３．３８（ｍ，１Ｈ），２．７１（ｄ，Ｊ＝５．４ Ｈｚ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４７９．０ ４８１．０［Ｍ＋Ｈ］^＋．ピーク２を減圧濃縮して残渣を得、これをＭｅＣＮ（１０ｍＬ）及びＨ_２Ｏ（２０ｍＬ）に溶解させ、凍結乾燥し、３－［（５Ｒ）－３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル］－Ｎ－メチル－４－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］ベンゼンスルホンアミド（４８．８１ｍｇ、１０１．８４μｍｏｌ、収率１４．１％、純度１００．０％、ＳＦＣ：Ｒ_ｔ＝１．９８２、ｅｅ＝１００％、［α］^３１．４ _Ｄ＝＋４０．０（ＣＨ_３ＯＨ、ｃ＝０．０５０ｇ／１００ｍＬ））を黄色固体として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．５１（ｓ，１Ｈ），８．２０（ｄ，Ｊ＝８．６ Ｈｚ，１Ｈ），７．８９（ｄｄ，Ｊ＝２．２，８．６ Ｈｚ，１Ｈ），７．８３－７．７６（ｍ，２Ｈ），７．３９（ｓ，１Ｈ），６．８１（ｄ，Ｊ＝８．８ Ｈｚ，１Ｈ），５．８１（ｔ，Ｊ＝１１．１ Ｈｚ，１Ｈ），４．３５（ｄ，Ｊ＝４．９ Ｈｚ，１Ｈ），３．６２－３．５１（ｍ，１Ｈ），３．４９－３．３９（ｍ，１Ｈ），２．７１（ｄ，Ｊ＝５．４ Ｈｚ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４７９．１，４８１．１［Ｍ＋Ｈ］^＋． Step 3: 3-[(5S)-3-bromo-4,5-dihydroisoxazol-5-yl]-N-methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]benzene sulfonamide, and 3-[(5R)-3-bromo-4,5-dihydroisoxazol-5-yl]-N-methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino] Benzene sulfonamide

N-methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]-3-vinyl-benzenesulfonamide (270 mg, 724.58 μmol, 95.9%, 1 equivalent) and dibromometa To a stirred solution of the non-oxime (293.94 mg, 1.45 mmol, 2 eq.) in EtOAc (10 mL) was added NaHCO ₃ (608.70 mg, 7.25 mmol, 281.80 μL, 10 eq.). The reaction mixture was stirred at 25°C for 2 hours. The reaction mixture was quenched by adding water (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Agela DuraShell C18 150*25 mm*5 μm; mobile phase : [Water (0.05% _NH3.H2O +10mM _NH4HCO3 )-ACN]; B%: 40%-70%, 10 _min ) and then lyophilized to obtain _a mixture, This was separated by a chiral SFC column (DAICEL CHIRALPAK IG (250 mm*30 mm, 10 μm); Mobile phase: [0.1% NH ₃ H ₂ O in MEOH]; B%: 40%-40%). Peak 1 and peak 2 were obtained. Peak 1 was concentrated in vacuo to give a residue, which was dissolved in MeCN (10 mL) and H ₂ O (20 mL) and lyophilized to give 3-[(5S)-3-bromo-4,5-dihydroisoxazole. -5-yl]-N-methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]benzenesulfonamide (44.51 mg, 92.87 μmol, yield 12.8%, purity 100. 0%, SFC: R _t = 1.658, ee = 100%, [α] ^31.4 _D = -44.4 (CH ₃ OH, c = 0.054 g/100 mL)) as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.51 (s, 1H), 8.19 (d, J = 8.6 Hz, 1H), 7.88 (dd, J = 2.1, 8 .7 Hz, 1H), 7.84-7.76 (m, 2H), 7.40 (s, 1H), 6.81 (d, J=8.8 Hz, 1H), 5.81 (t , J=11.1 Hz, 1H), 4.41 (s, 1H), 3.62-3.52 (m, 1H), 3.49-3.38 (m, 1H), 2.71 ( d, J=5.4 Hz, 3H); ES-LCMS m/z 479.0 481.0 [M+H] ⁺ . Peak 2 was concentrated under reduced pressure to give a residue, which was dissolved in MeCN (10 mL) and H ₂ O (20 mL) and lyophilized to give 3-[(5R)-3-bromo-4,5-dihydroisoxazole. -5-yl]-N-methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]benzenesulfonamide (48.81 mg, 101.84 μmol, yield 14.1%, purity 100. 0%, SFC: R _t = 1.982, ee = 100%, [α] ^31.4 _D = +40.0 (CH ₃ OH, c = 0.050 g/100 mL)) as a yellow solid. ^1H NMR (400 MHz, _CDCl3 ) δ ppm 8.51 (s, 1H), 8.20 (d, J = 8.6 Hz, 1H), 7.89 (dd, J = 2.2, 8 .6 Hz, 1H), 7.83-7.76 (m, 2H), 7.39 (s, 1H), 6.81 (d, J=8.8 Hz, 1H), 5.81 (t , J = 11.1 Hz, 1H), 4.35 (d, J = 4.9 Hz, 1H), 3.62-3.51 (m, 1H), 3.49-3.39 (m, 1H), 2.71 (d, J=5.4 Hz, 3H); ES-LCMS m/z 479.1, 481.1 [M+H] ⁺ .

５－ブロモ－２－ヨード－アニリン（２．５ｇ、８．３９ｍｍｏｌ、１当量）及び４－（トリフルオロメチル）ベンズアルデヒド（４．３８ｇ、２５．１７ｍｍｏｌ、３．３７ｍＬ、３当量）の、ＭｅＯＨ（２５ｍＬ）溶液に、ＡｃＯＨ（５０．３９ｍｇ、８３９．１６μｍｏｌ、４７．９９μＬ、０．１当量）を加えた。混合物を６０℃で４時間撹拌した。ＮａＢＨ_３ＣＮ（２．６４ｇ、４１．９６ｍｍｏｌ、５当量）を２５℃で加えた。混合物を６０℃で１２時間撹拌した。溶媒を除去し、水（２００ｍＬ）を加えて残渣をクエンチして、ＥｔＯＡｃで抽出した（１００ｍＬ×３）。合わせた有機層をブライン（５０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、これをフラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１／０から５／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝５／１、Ｒ_ｆ＝０．７０）により精製し、５－ブロモ－２－ヨード－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］アニリン（１．８８ｇ、３．６７ｍｍｏｌ、収率４３．７％、純度８９．０％）を黄色油として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ７．６４（ｄ，Ｊ＝８．１ Ｈｚ，２Ｈ），７．５２（ｄ，Ｊ＝８．３ Ｈｚ，１Ｈ），７．４７（ｄ，Ｊ＝７．８ Ｈｚ，２Ｈ），６．６４－６．５８（ｍ，２Ｈ），４．７２（ｓ，１Ｈ），４．４７（ｄ，Ｊ＝５．６ Ｈｚ，２Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４５５．９，４５７．９［Ｍ＋Ｈ］^＋． I-12 and I-13 (isomers of I-26)
Step 1: 5-bromo-2-iodo-N-[[4-(trifluoromethyl)phenyl]methyl]aniline

5-bromo-2-iodo-aniline (2.5 g, 8.39 mmol, 1 eq.) and 4-(trifluoromethyl)benzaldehyde (4.38 g, 25.17 mmol, 3.37 mL, 3 eq.) in MeOH ( AcOH (50.39 mg, 839.16 μmol, 47.99 μL, 0.1 eq.) was added to the 25 mL) solution. The mixture was stirred at 60°C for 4 hours. _NaBH3CN (2.64g, 41.96mmol, 5eq) was added at 25<0>C. The mixture was stirred at 60°C for 12 hours. The solvent was removed and the residue was quenched by adding water (200 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give a residue, which was subjected to flash silica gel chromatography (PE/EtOAc = 1/0 to 5/1; TLC: PE/EtOAc=5/1, R _f =0.70) to give 5-bromo-2-iodo-N-[[4-(trifluoromethyl)phenyl]methyl]aniline (1.88 g, 3.67 mmol, yield 43.7%, purity 89.0%) was obtained as a yellow oil. ^1H NMR (400 MHz, _CDCl3 ) δ ppm 7.64 (d, J=8.1 Hz, 2H), 7.52 (d, J=8.3 Hz, 1H), 7.47 (d, ES- LCMS m/z 455.9, 457.9 [M+H] ⁺ .

５－ブロモ－２－ヨード－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］アニリン（１．８８ｇ、３．６７ｍｍｏｌ、８９％、１当量）のＴＨＦ（２０ｍＬ）溶液に、ＤＭＡＰ（４４８．２４ｍｇ、３．６７ｍｍｏｌ、１当量）及びＢｏｃ_２Ｏ（２．４０ｇ、１１．０１ｍｍｏｌ、２．５３ｍＬ、３当量）を加えた。混合物を２０℃で１２時間撹拌した。溶媒を除去して残渣を得、これをフラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１／０から１０／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝１０／１、Ｒ_ｆ＝０．８）により精製して、ｔｅｒｔ－ブチル＝Ｎ－（５－ブロモ－２－ヨード－フェニル）－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］カルバマート（２．２３ｇ、粗）を緑色固体として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ７．７１（ｄ，Ｊ＝８．３ Ｈｚ，１Ｈ），７．５９（ｄ，Ｊ＝７．３ Ｈｚ，２Ｈ），７．３８（ｄ，Ｊ＝７．８ Ｈｚ，２Ｈ），７．１２（ｄ，Ｊ＝７．８ Ｈｚ，１Ｈ），６．９７（ｓ，１Ｈ），５．１６（ｄ，Ｊ＝１５．２ Ｈｚ，１Ｈ），４．２７（ｄ，Ｊ＝１４．９ Ｈｚ，１Ｈ），１．５７（ｓ，９Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４９９．９，４２１．９［Ｍ－ｔ－Ｂｕ＋Ｈ］^＋． Step 2: tert-butyl=N-(5-bromo-2-iodo-phenyl)-N-[[4-(trifluoromethyl)phenyl]methyl]carbamate

DMAP (448 .24 mg, 3.67 mmol, 1 eq) and _Boc2O (2.40 g, 11.01 mmol, 2.53 mL, 3 eq) were added. The mixture was stirred at 20°C for 12 hours. Removal of the solvent gave a residue, which was purified by flash silica gel chromatography (PE/EtOAc=1/0 to 10/1, TLC: PE/EtOAc=10/1, R _f =0.8). tert-Butyl N-(5-bromo-2-iodo-phenyl)-N-[[4-(trifluoromethyl)phenyl]methyl]carbamate (2.23 g, crude) was obtained as a green solid. ^1H NMR (400 MHz, _CDCl3 ) δ ppm 7.71 (d, J=8.3 Hz, 1H), 7.59 (d, J=7.3 Hz, 2H), 7.38 (d, J=7.8 Hz, 2H), 7.12 (d, J=7.8 Hz, 1H), 6.97 (s, 1H), 5.16 (d, J=15.2 Hz, 1H) , 4.27 (d, J=14.9 Hz, 1H), 1.57 (s, 9H); ES-LCMS m/z 499.9, 421.9 [M-t-Bu+H] ⁺ .

ｔｅｒｔ－ブチル＝Ｎ－（５－ブロモ－２－ヨード－フェニル）－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］カルバマート（１ｇ、１．７３ｍｍｏｌ、純度９６．３％、１当量）、トリブチル－（１－メチルイミダゾール－４－イル）スタンナン（６６２．５３ｍｇ、１．７３ｍｍｏｌ、純度９７％、１当量）、及びＰｄ（ｄｐｐｆ）Ｃｌ_２（１２６．７０ｍｇ、１７３．１５μｍｏｌ、０．１当量）の、ＤＭＦ（１０ｍＬ）との混合物を脱気し、Ｎ_２で３回パージして、混合物をＮ_２雰囲気下、１２０℃で４時間撹拌した。水（１００ｍＬ）を添加することにより反応混合物をクエンチし、ＥｔＯＡｃで抽出した（５０ｍＬ×３）。合わせた有機層をブライン（２０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、フラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１／０から３／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝３／１、Ｒ_ｆ＝０．４０）により精製し、ｔｅｒｔ－ブチル＝Ｎ－［５－ブロモ－２－（１－メチルイミダゾール－４－イル）フェニル］－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］カルバマート（５３０ｍｇ、９３４．６６μｍｏｌ、収率５４．０％、純度９０．０％）を黒褐色油として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ７．９７（ｄ，Ｊ＝７．６ Ｈｚ，１Ｈ），７．５５（ｄ，Ｊ＝８．１ Ｈｚ，２Ｈ），７．４６（ｓ，２Ｈ），７．３５（ｄ，Ｊ＝６．８ Ｈｚ，２Ｈ），６．９４（ｓ，１Ｈ），６．８２（ｓ，１Ｈ），５．１５（ｄ，Ｊ＝１４．４ Ｈｚ，１Ｈ），４．１８（ｄ，Ｊ＝１４．９ Ｈｚ，１Ｈ），３．６６（ｓ，３Ｈ），１．２６（ｓ，９Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ５１０．１，５１２．１ ［Ｍ＋Ｈ］^＋． Step 3: tert-butyl=N-[5-bromo-2-(1-methylimidazol-4-yl)phenyl]-N-[[4-(trifluoromethyl)phenyl]methyl]carbamate

tert-Butyl N-(5-bromo-2-iodo-phenyl)-N-[[4-(trifluoromethyl)phenyl]methyl]carbamate (1 g, 1.73 mmol, purity 96.3%, 1 equivalent) , tributyl-(1-methylimidazol-4-yl)stannane (662.53 mg, 1.73 mmol, purity 97%, 1 eq.), and Pd(dppf)Cl ₂ (126.70 mg, 173.15 μmol, 0.1 (eq.) with DMF (10 mL) was degassed and purged with _N2 three times, and the mixture was stirred at 120 °C under _N2 atmosphere for 4 h. The reaction mixture was quenched by adding water (100 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give a residue, which was subjected to flash silica gel chromatography (PE/EtOAc = 1/0 to 3/1, TLC: tert-butyl N-[5-bromo-2-(1-methylimidazol-4-yl)phenyl] _-N -[[4 -(trifluoromethyl)phenyl]methyl]carbamate (530 mg, 934.66 μmol, yield 54.0%, purity 90.0%) was obtained as a dark brown oil. ^1H NMR (400 MHz, _CDCl3 ) δ ppm 7.97 (d, J=7.6 Hz, 1H), 7.55 (d, J=8.1 Hz, 2H), 7.46 (s, 2H), 7.35 (d, J = 6.8 Hz, 2H), 6.94 (s, 1H), 6.82 (s, 1H), 5.15 (d, J = 14.4 Hz, 1H), 4.18 (d, J = 14.9 Hz, 1H), 3.66 (s, 3H), 1.26 (s, 9H); ES-LCMS m/z 510.1, 512.1 [M+H] ⁺ .

Step 4: tert-butyl=N-[2-(1-methylimidazol-4-yl)-5-vinyl-phenyl]-N-[[4-(trifluoromethyl)phenyl]methyl]carbamate
tert-Butyl N-[5-bromo-2-(1-methylimidazol-4-yl)phenyl]-N-[[4-(trifluoromethyl)phenyl]methyl]carbamate (530 mg, 934.66 μmol, 90 %, 1 eq), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (287.90 mg, 1.87 mmol, 317.07 μL, 2 eq), Pd(dppf)Cl ₂ (68.39 mg, 93.47 μmol, 0.1 eq), Cs ₂ CO ₃ (761.33 mg, 2.34 mmol, 2.5 eq), 1,4-dioxane (4.5 mL) and H ₂ O ( 1.5 mL) was degassed and purged with _N2 three times, and the mixture was stirred at 100 °C under _N2 atmosphere for 12 h. The reaction mixture was quenched by adding water (50 mL) and extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine (10 _mL ), dried over _Na2SO4 , filtered and concentrated in vacuo to give a residue, which was purified by flash silica gel chromatography (fPE/EtOAc=1/0 to 3/1, TLC: tert-butyl N-[2-(1-methylimidazol-4-yl)-5-vinyl-phenyl] _-N -[[ 4-(Trifluoromethyl)phenyl]methyl]carbamate (219 mg, 464.34 μmol, yield 49.7%, purity 97.0%) was obtained as a yellow oil. ^1H NMR (400 MHz, _CDCl3 ) δ ppm 8.05 (d, J = 8.1 Hz, 1H), 7.54 (d, J = 7.8 Hz, 3H), 7.47 (s, 1H), 7.38 (d, J = 7.3 Hz, 3H), 6.85 (s, 1H), 6.74 (s, 1H), 6.57 (dd, J = 10.9, 17 .5 Hz, 1H), 5.55 (d, J=17.4 Hz, 1H), 5.18 (d, J=10.8 Hz, 1H), 5.22-5.15 (m, 1H) ), 3.67 (s, 3H), 1.25 (s, 9H); ES-LCMS m/z 458.1 [M+H] ⁺ .

ｔｅｒｔ－ブチルＮ－［２－（１－メチルイミダゾール－４－イル）－５－ビニル－フェニル］－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］カルバマート（１６９ｍｇ、３５８．３３μｍｏｌ、９７％、１当量）、及びジブロモメタノンオキシム（１０９．０２ｍｇ、５３７．４９μｍｏｌ、１．５当量）の、ＥｔＯＡｃ（３ｍＬ）溶液に、ＮａＨＣＯ_３（３０１．０２ｍｇ、３．５８ｍｍｏｌ、１０当量）を加えた。混合物を２５℃で４時間撹拌した。水（５０ｍＬ）を添加することにより反応混合物をクエンチし、ＥｔＯＡｃで抽出した（３０ｍＬ×３）。合わせた有機層をブライン（１０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して、ｔｅｒｔ－ブチル＝Ｎ－［５－（３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル）－２－（１－メチルイミダゾール－４－イル）フェニル］－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］カルバマート（２４８ｍｇ、粗）を黄色固体として得、これをさらに精製することなく、次工程で使用した。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．１０（ｓ，１Ｈ），７．６４（ｓ，２Ｈ），７．６０（ｄ，Ｊ＝８．３ Ｈｚ，２Ｈ），７．５１－７．４７（ｍ，１Ｈ），６．９５－６．８２（ｍ，２Ｈ），６．６８（ｓ，１Ｈ），５．５４（ｄ，Ｊ＝８．８ Ｈｚ，１Ｈ），５．３０－５．０８（ｍ，２Ｈ），３．７１（ｓ，３Ｈ），３．５９－３．４５（ｍ，２Ｈ），１．５４（ｓ，９Ｈ）． Step 5: tert-butyl N-[2-(1-methylimidazol-4-yl)-5-vinyl-phenyl]-N-[[4-(trifluoromethyl)phenyl]methyl]carbamate

tert-Butyl N-[2-(1-methylimidazol-4-yl)-5-vinyl-phenyl]-N-[[4-(trifluoromethyl)phenyl]methyl]carbamate (169 mg, 358.33 μmol, 97 %, 1 eq.) and dibromomethanone oxime (109.02 mg, 537.49 μmol, 1.5 eq.) in EtOAc (3 mL) was added NaHCO ₃ (301.02 mg, 3.58 mmol, 10 eq.). Ta. The mixture was stirred at 25°C for 4 hours. The reaction mixture was quenched by adding water (50 mL) and extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine (10 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give tert-butyl N-[5-(3-bromo-4,5-dihydroisoxazole-). 5-yl)-2-(1-methylimidazol-4-yl)phenyl]-N-[[4-(trifluoromethyl)phenyl]methyl]carbamate (248 mg, crude) was obtained as a yellow solid, which was further It was used in the next step without purification. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.10 (s, 1H), 7.64 (s, 2H), 7.60 (d, J = 8.3 Hz, 2H), 7.51- 7.47 (m, 1H), 6.95-6.82 (m, 2H), 6.68 (s, 1H), 5.54 (d, J=8.8 Hz, 1H), 5.30 -5.08 (m, 2H), 3.71 (s, 3H), 3.59-3.45 (m, 2H), 1.54 (s, 9H).

ｔｅｒｔ－ブチル Ｎ－［５－（３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル）－２－（１－メチルイミダゾール－４－イル）フェニル］－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］カルバマート（２４８ｍｇ、４２８．０２μｍｏｌ、１当量）の、ＤＣＭ（３ｍＬ）溶液に、ＴＦＡ（１８４．８０ｍｇ、１．６２ｍｍｏｌ、１２０μＬ、３．７９当量）を加えた。混合物を２５℃で１時間撹拌した。水（１００ｍＬ）を添加することにより反応混合物をクエンチし、ＥｔＯＡｃで抽出した（６０ｍＬ×３）。合わせた有機層をブライン（２０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、フラッシュシリカゲルクロマトグラフィー（ｆＰＥ／ＥｔＯＡｃ＝１００／１から３／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝３／１、Ｒ_ｆ＝０．０５）により精製して化合物を得、これをＳＦＣ（カラム：ＤＡＩＣＥＬ ＣＨＩＲＡＬＰＡＫ ＡＤ（２５０ｍｍ＊３０ｍｍ、１０μｍ）；移動相：［０．１％のＮＨ_３Ｈ_２Ｏ ＥＴＯＨ］；Ｂ％：５０％－５０％）により分離して、ピーク１及びピーク２を得た。ピーク１を、分取ＨＰＬＣ（カラム：Ａｇｅｌａ ＤｕｒａＳｈｅｌｌ Ｃ１８ １５０＊２５ｍｍ＊５μｍ；移動相：［水（０．０５％のＮＨ_３Ｈ_２Ｏ＋１０ｍＭ ＮＨ_４ＨＣＯ_３）－ＡＣＮ］；Ｂ％：５２％－８２％、１０分）により精製して凍結乾燥し、（Ｓ）－５－（３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル）－２－（１－メチル－１Ｈ－イミダゾール－４－イル）－Ｎ－（４－（トリフルオロメチル）ベンジル）アニリン（１０．４４ｍｇ、２１．３５μｍｏｌ、収率５．０％、純度９８．５％、ＳＦＣ：Ｒ_ｔ＝２．１４３、ｅｅ＝９８．２％、［α］^３２．０ _Ｄ＝＋１８０．０（ＭｅＯＨ、ｃ＝０．０２ｇ／１００ｍＬ））を白色固体として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．５２（ｓ，１Ｈ），７．６０－７．５６（ｍ，２Ｈ），７．５４－７．５０（ｍ，２Ｈ），７．４７（ｓ，１Ｈ），７．３９（ｄ，Ｊ＝７．８ Ｈｚ，１Ｈ），７．１８（ｓ，１Ｈ），６．６１（ｄ，Ｊ＝７．８ Ｈｚ，１Ｈ），６．４７（ｓ，１Ｈ），５．５７－５．４７（ｍ，１Ｈ），４．５５（ｓ，２Ｈ），３．７７（ｓ，３Ｈ），３．５０（ｄｄ，Ｊ＝１０．９，１７．２ Ｈｚ，１Ｈ），３．０７（ｄｄ，Ｊ＝８．９，１７．２ Ｈｚ，１Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４７８．８，４８０．８［Ｍ＋Ｈ］^＋．ピーク２を、分取ＨＰＬＣ（カラム：Ａｇｅｌａ ＤｕｒａＳｈｅｌｌ Ｃ１８ １５０＊２５ｍｍ＊５μｍ；移動相：［水（０．０５％のＮＨ_３Ｈ_２Ｏ＋１０ｍＭ ＮＨ_４ＨＣＯ_３）－ＡＣＮ］；Ｂ％：５１％－８１％、１０分）により精製して凍結乾燥し、（Ｒ）－５－（３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル）－２－（１－メチル－１Ｈ－イミダゾール－４－イル）－Ｎ－（４－（トリフルオロメチル）ベンジル）アニリン（９．５ｍｇ、１８．９９μｍｏｌ、収率４．４％、純度９５．９％、ＳＦＣ：Ｒ_ｔ＝２．４８２、ｅｅ＝９８．８％、［α］^３２．０ _Ｄ＝－１８７．５（ＭｅＯＨ、ｃ＝０．０１６ｇ／１００ｍＬ））を白色固体として得た。^１Ｈ ＮＭＲ （４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．５２（ｓ，１Ｈ），７．６０－７．５６（ｍ，２Ｈ），７．５３－７．５０（ｍ，２Ｈ），７．４７（ｓ，１Ｈ），７．３９（ｄ，Ｊ＝７．８ Ｈｚ，１Ｈ），７．１８（ｄ，Ｊ＝１．２ Ｈｚ，１Ｈ），６．６１（ｄ，Ｊ＝７．８ Ｈｚ，１Ｈ），６．４７（ｓ，１Ｈ），５．５２（ｄｄ，Ｊ＝８．９，１０．９ Ｈｚ，１Ｈ），４．５５（ｓ，２Ｈ），３．７７（ｓ，３Ｈ），３．５０（ｄｄ，Ｊ＝１０．９，１７．２ Ｈｚ，１Ｈ），３．０７（ｄｄ，Ｊ＝８．８，１７．１ Ｈｚ，１Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４７９．０，４８１．０［Ｍ＋Ｈ］^＋． Step 6: (S)-5-(3-bromo-4,5-dihydroisoxazol-5-yl)-2-(1-methyl-1H-imidazol-4-yl)-N-(4-(tri fluoromethyl)benzyl)aniline, and (R)-5-(3-bromo-4,5-dihydroisoxazol-5-yl)-2-(1-methyl-1H-imidazol-4-yl)-N- (4-(trifluoromethyl)benzyl)aniline

tert-Butyl N-[5-(3-bromo-4,5-dihydroisoxazol-5-yl)-2-(1-methylimidazol-4-yl)phenyl]-N-[[4-(trifluoro To a solution of methyl)phenyl]methyl]carbamate (248 mg, 428.02 μmol, 1 eq.) in DCM (3 mL) was added TFA (184.80 mg, 1.62 mmol, 120 μL, 3.79 eq.). The mixture was stirred at 25°C for 1 hour. The reaction mixture was quenched by adding water (100 mL) and extracted with EtOAc (60 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give a residue, which was subjected to flash silica gel chromatography (fPE/EtOAc = 100/1 to 3/1, TLC: PE/EtOAc=3/1, R _f =0.05) to obtain a compound, which was purified by SFC (column: DAICEL CHIRALPAK AD (250 mm * 30 mm, 10 μm); mobile phase: [0.1% NH ₃ H ₂ O ETOH]; B%: 50%-50%) to obtain peak 1 and peak 2. Peak 1 was analyzed by preparative HPLC (column: Agela DuraShell C18 150*25mm*5μm; mobile phase: [water (0.05% NH ₃ H ₂ O + 10mM NH ₄ HCO ₃ )-ACN]; B%: 52%- (82%, 10 min) and lyophilized to give (S)-5-(3-bromo-4,5-dihydroisoxazol-5-yl)-2-(1-methyl-1H-imidazol-4 -yl)-N-(4-(trifluoromethyl)benzyl)aniline (10.44 mg, 21.35 μmol, yield 5.0%, purity 98.5%, SFC: R _t =2.143, ee= 98.2%, [α] ^32.0 _D = +180.0 (MeOH, c = 0.02 g/100 mL)) was obtained as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.52 (s, 1H), 7.60-7.56 (m, 2H), 7.54-7.50 (m, 2H), 7.47 (s, 1H), 7.39 (d, J = 7.8 Hz, 1H), 7.18 (s, 1H), 6.61 (d, J = 7.8 Hz, 1H), 6.47 (s, 1H), 5.57-5.47 (m, 1H), 4.55 (s, 2H), 3.77 (s, 3H), 3.50 (dd, J=10.9, 17 .2 Hz, 1H), 3.07 (dd, J=8.9, 17.2 Hz, 1H); ES-LCMS m/z 478.8, 480.8 [M+H] ⁺ . Peak 2 was purified by preparative HPLC (column: Agela DuraShell C18 150*25mm*5μm; mobile phase: [water (0.05% NH ₃ H ₂ O + 10mM NH ₄ HCO ₃ )-ACN]; B%: 51%- (81%, 10 min) and lyophilized to give (R)-5-(3-bromo-4,5-dihydroisoxazol-5-yl)-2-(1-methyl-1H-imidazol-4 -yl)-N-(4-(trifluoromethyl)benzyl)aniline (9.5 mg, 18.99 μmol, yield 4.4%, purity 95.9%, SFC: R _t =2.482, ee= 98.8%, [α] ^32.0 _D = -187.5 (MeOH, c = 0.016 g/100 mL)) was obtained as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.52 (s, 1H), 7.60-7.56 (m, 2H), 7.53-7.50 (m, 2H), 7.47 (s, 1H), 7.39 (d, J = 7.8 Hz, 1H), 7.18 (d, J = 1.2 Hz, 1H), 6.61 (d, J = 7.8 Hz) , 1H), 6.47 (s, 1H), 5.52 (dd, J=8.9, 10.9 Hz, 1H), 4.55 (s, 2H), 3.77 (s, 3H) , 3.50 (dd, J = 10.9, 17.2 Hz, 1H), 3.07 (dd, J = 8.8, 17.1 Hz, 1H); ES-LCMS m/z 479.0 , 481.0 [M+H] ⁺ .

５－（トリフルオロメチル）ピリジン－２－アミン（１．８４ｇ、１１．３６ｍｍｏｌ、１当量）のＴＨＦ（３０ｍＬ）溶液に、ＮａＨ（１．３６ｇ、３４．０９ｍｍｏｌ、６０％、３当量）を０℃で加えた。混合物を３０分間撹拌した。２－ブロモ－１－フルオロ－４－ニトロ－ベンゼン（２．５ｇ、１１．２６ｍｍｏｌ、１当量）を０℃で加え、混合物を２５℃で１２時間撹拌した。反応混合物を水（５０ｍＬ）でクエンチし、ＥｔＯＡｃで抽出した（５０ｍＬ×３）。合わせた有機層をＮａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、これをフラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１００／１から１０／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝５／１、Ｒ_ｆ＝０．６５）により精製し、Ｎ－（２－ブロモ－４－ニトロ－フェニル）－５－（トリフルオロメチル）ピリジン－２－アミン（８５０ｍｇ、２．００ｍｍｏｌ、収率１７．６％、純度８５．０％）を黄色固体として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．７６（ｄ，Ｊ＝９．３ Ｈｚ，１Ｈ），８．６３（ｓ，１Ｈ），８．５２（ｄ，Ｊ＝２．４ Ｈｚ，１Ｈ），８．２４（ｄｄ，Ｊ＝２．４，９．３ Ｈｚ，１Ｈ），７．８７（ｄｄ，Ｊ＝２．２，８．８ Ｈｚ，１Ｈ），７．５４ （ｓ，１Ｈ），７．００（ｄ，Ｊ＝８．８ Ｈｚ，１Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ３６３．９［Ｍ＋Ｈ］^＋． I-14
Step 1: N-(2-bromo-4-nitrophenyl)-5-(trifluoromethyl)pyridin-2-amine

To a solution of 5-(trifluoromethyl)pyridin-2-amine (1.84 g, 11.36 mmol, 1 eq.) in THF (30 mL) was added 0% NaH (1.36 g, 34.09 mmol, 60%, 3 eq.). Added at °C. The mixture was stirred for 30 minutes. 2-bromo-1-fluoro-4-nitro-benzene (2.5 g, 11.26 mmol, 1 eq.) was added at 0° C. and the mixture was stirred at 25° C. for 12 hours. The reaction mixture was quenched with water (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was subjected to flash silica gel chromatography (PE/EtOAc = 100/1 to 10/1, TLC: PE/EtOAc = 5/ 1, R _f =0.65) to give N-(2-bromo-4-nitro-phenyl)-5-(trifluoromethyl)pyridin-2-amine (850 mg, 2.00 mmol, yield 17. 6%, purity 85.0%) was obtained as a yellow solid. ^1H NMR (400 MHz, _CDCl3 ) δ ppm 8.76 (d, J = 9.3 Hz, 1H), 8.63 (s, 1H), 8.52 (d, J = 2.4 Hz, 1H), 8.24 (dd, J=2.4, 9.3 Hz, 1H), 7.87 (dd, J=2.2, 8.8 Hz, 1H), 7.54 (s, 1H ), 7.00 (d, J=8.8 Hz, 1H); ES-LCMS m/z 363.9 [M+H] ⁺ .

Ｎ－（２－ブロモ－４－ニトロ－フェニル）－５－（トリフルオロメチル）ピリジン－２－アミン（７００ｍｇ、１．９３ｍｍｏｌ、１当量）及びトリブチル－（１－メチルイミダゾール－４－イル）スタンナン（１．５８ｇ、３．８７ｍｍｏｌ、９１％、２当量）の、ＤＭＦ（１５ｍＬ）溶液に、Ｐｄ（ｄｐｐｆ）Ｃｌ_２（７０．７３ｍｇ、９６．６６μｍｏｌ、０．０５当量）をＮ_２雰囲気下で加えた。混合物を、Ｎ_２雰囲気下、１３０℃で１２時間撹拌した。反応混合物を水（５０ｍＬ）とＥｔＯＡｃ（１００ｍＬ×３）とに分配した。有機相を分離し、ブライン（５０ｍＬ）で洗浄してＮａ_２ＳＯ_４で乾燥させ、濾過し、減圧濃縮して残渣を得、これをシリカゲルカラムクロマトグラフィー（純粋なＰＥから、ＰＥ／ＥｔＯＡｃ＝０／１まで、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝０／１、Ｒ_ｆ＝０．３４）により精製し、２－（１－メチルイミダゾール－４－イル）－Ｎ^１－［５－（トリフルオロメチル）－２－ピリジル］ベンゼン－１，４－ジアミン（３００ｍｇ、６４８．０４μｍｏｌ、収率３３．５％、純度７２．１％）を黄色油として得た。^１Ｈ ＮＭＲ （４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ９．４１（ｓ，１Ｈ），８．４０（ｓ，１Ｈ），７．７２（ｄ，Ｊ＝８．６ Ｈｚ，１Ｈ），７．５９－７．４３（ｍ，２Ｈ），７．２７（ｓ，１Ｈ），７．１８－７．０１（ｍ，２Ｈ），６．７４－６．５７（ｍ，２Ｈ），３．７１（ｓ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ３３４．３［Ｍ＋Ｈ］^＋． Step 2: 2-(1-methyl-1H-imidazol-4-yl)-N ¹ -(5-(trifluoromethyl)pyridin-2-yl)benzene-1,4-diamine

N-(2-bromo-4-nitro-phenyl)-5-(trifluoromethyl)pyridin-2-amine (700 mg, 1.93 mmol, 1 eq) and tributyl-(1-methylimidazol-4-yl)stannane (1.58 g, 3.87 mmol, 91%, 2 eq.) in DMF (15 mL) was added Pd(dppf) _Cl2 (70.73 mg, 96.66 μmol, 0.05 eq.) under _N2 atmosphere. added. The mixture was stirred at 130 °C for 12 h under _N2 atmosphere. The reaction mixture was partitioned between water (50 mL) and EtOAc (100 mL x 3). The organic phase was separated, washed with brine (50 mL), dried over _Na2SO4 , filtered, and concentrated _under reduced pressure to give a residue, which was purified by silica gel column chromatography (from pure PE, PE/EtOAc=0 2-(1-methylimidazol-4-yl)-N ¹ -[ _5- (trifluoromethyl)- 2-Pyridyl]benzene-1,4-diamine (300 mg, 648.04 μmol, 33.5% yield, 72.1% purity) was obtained as a yellow oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.41 (s, 1H), 8.40 (s, 1H), 7.72 (d, J = 8.6 Hz, 1H), 7.59- 7.43 (m, 2H), 7.27 (s, 1H), 7.18-7.01 (m, 2H), 6.74-6.57 (m, 2H), 3.71 (s, 3H); ES-LCMS m/z 334.3 [M+H] ⁺ .

２－（１－メチルイミダゾール－４－イル）－Ｎ^１－［５－（トリフルオロメチル）－２－ピリジル］ベンゼン－１，４－ジアミン（２３０ｍｇ、４９６．８３μｍｏｌ、７２％、１当量）及びＥｔ_３Ｎ（１５０．８２ｍｇ、１．４９ｍｍｏｌ、２０７．４６μＬ、３当量）の、ＤＣＭ（３ｍＬ）溶液に、プロパ－２－エノイルクロリド（５８．４６ｍｇ、６４５．８８μｍｏｌ、５２．６６μＬ、１．３当量）をＮ_２雰囲気下、０℃で加えた。混合物を２５℃で２時間撹拌した。反応混合物を水（３０ｍＬ）とＥｔＯＡｃ（５０ｍＬ×３）とに分配した。有機相を分離し、ブライン（３０ｍＬ）で洗浄してＮａ_２ＳＯ_４で乾燥させ、濾過し、減圧濃縮して残渣を得、これを分取ＨＰＬＣ（カラム：Ａｇｅｌａ ＤｕｒａＳｈｅｌｌ Ｃ１８ １５０＊２５ｍｍ＊５μｍ；移動相：［水（０．０５％のＮＨ_３Ｈ_２Ｏ＋１０ｍＭのＮＨ_４ＨＣＯ_３）－ＡＣＮ］；Ｂ％：３８％－６８％、１０分）により精製して、Ｎ－［３－（１－メチルイミダゾール－４－イル）－４－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］フェニル］プロパ－２－エナミド（８５．１５ｍｇ、２１５．４３μｍｏｌ、収率４３．４％、純度９８．２％）を白色固体として得た。^１Ｈ ＮＭＲ（５００ ＭＨｚ， ＣＤＣｌ_３）δ ｐｐｍ １１．３０（ｓ，１Ｈ），８．４４（ｓ，１Ｈ），８．３８（ｄ，Ｊ＝８．９ Ｈｚ，１Ｈ），８．１７（ｄ，Ｊ＝２．０ Ｈｚ，１Ｈ），７．７４（ｓ，１Ｈ），７．６０（ｄｄ，Ｊ＝２．０，８．９ Ｈｚ，１Ｈ），７．４７（ｓ，１Ｈ），７．１８（ｓ，２Ｈ），６．８２（ｄ，Ｊ＝８．７ Ｈｚ，１Ｈ），６．４９－６．４０（ｍ，１Ｈ），６．３５－６．２４（ｍ，１Ｈ），５．７５（ｄ，Ｊ＝１０．４ Ｈｚ，１Ｈ），３．６８（ｓ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ３８８．２［Ｍ＋Ｈ］^＋． Step 3: N-(3-(1-methyl-1H-imidazol-4-yl)-4-((5-(trifluoromethyl)pyridin-2-yl)amino)phenyl)acrylamide

2-(1-methylimidazol-4-yl)-N ¹ -[5-(trifluoromethyl)-2-pyridyl]benzene-1,4-diamine (230 mg, 496.83 μmol, 72%, 1 equivalent) and To a solution of Et ₃ N (150.82 mg, 1.49 mmol, 207.46 μL, 3 eq.) in DCM (3 mL) was added prop-2-enoyl chloride (58.46 mg, 645.88 μmol, 52.66 μL, 1. (3 eq.) was added at 0 °C under _N2 atmosphere. The mixture was stirred at 25°C for 2 hours. The reaction mixture was partitioned between water (30 mL) and EtOAc (50 mL x 3). The organic phase was separated, washed with brine (30 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Agela DuraShell C18 150*25 mm*5 μm; Mobile phase: [Water (0.05% NH ₃ H ₂ O + 10 mM NH ₄ HCO ₃ )-ACN]; B%: 38%-68%, 10 min) to purify N-[3-(1 -methylimidazol-4-yl)-4-[[5-(trifluoromethyl)-2-pyridyl]amino]phenyl]prop-2-enamide (85.15 mg, 215.43 μmol, yield 43.4%, (purity 98.2%) was obtained as a white solid. ^1H NMR (500 MHz, _CDCl3 ) δ ppm 11.30 (s, 1H), 8.44 (s, 1H), 8.38 (d, J = 8.9 Hz, 1H), 8.17 ( d, J=2.0 Hz, 1H), 7.74 (s, 1H), 7.60 (dd, J=2.0, 8.9 Hz, 1H), 7.47 (s, 1H), 7.18 (s, 2H), 6.82 (d, J=8.7 Hz, 1H), 6.49-6.40 (m, 1H), 6.35-6.24 (m, 1H) , 5.75 (d, J=10.4 Hz, 1H), 3.68 (s, 3H); ES-LCMS m/z 388.2 [M+H] ⁺ .

工程１：３－［（５Ｓ）－３－クロロ－４，５－ジヒドロイソオキサゾール－５－イル］－Ｎ－メチル－４－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］ベンゼンスルホンアミド

３－（３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル）－Ｎ－メチル－４－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］ベンゼンスルホンアミド（９００ｍｇ、７２３．３５μｍｏｌ、純度３８．５％、１当量）の１，４－ジオキサン（１０ｍＬ）溶液に、塩酸（２ｍＬ）を加えた。混合物を４０℃で１２時間撹拌した。飽和ＮａＨＣＯ_３（１００ｍＬ）を添加することにより反応混合物をクエンチし、ＥｔＯＡｃで抽出した（５０ｍＬ×３）。合わせた有機層をブライン（５０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、これを分取ＨＰＬＣ（カラム：Ａｇｅｌａ ＤｕｒａＳｈｅｌｌ Ｃ_１８ １５０＊２５ｍｍ＊５μｍ；移動相：［水（０．０５％のＮＨ_３・Ｈ_２Ｏ＋１０ｍＭのＮＨ_４ＨＣＯ_３）－ＡＣＮ］；Ｂ％：４０％－７０％、１０分）により精製し、次いで凍結乾燥して混合物を得、これをキラルＳＦＣカラム（カラム：カラム：ＤＡＩＣＥＬ ＣＨＩＲＡＬＰＡＫ ＡＳ（２５０ｍｍ＊３０ｍｍ、１０μｍ）；移動相：［０．１％のＮＨ_３・Ｈ_２ＯのＥｔＯＨ］；Ｂ％：２５％－２０％）により分離して、ピーク１及びピーク２を得た。ピーク１を減圧濃縮して残渣を得、これをＭｅＣＮ（１０ｍＬ）及びＨ_２Ｏ（２０ｍＬ）に溶解させた後、凍結乾燥させ、３－［（５Ｓ）－３－クロロ－４，５－ジヒドロイソオキサゾール－５－イル］－Ｎ－メチル－４－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］ベンゼンスルホンアミド（４８．３６ｍｇ、１１１．２２μｍｏｌ、収率１５．４％、純度１００．０％、ＳＦＣ：Ｒ_ｔ＝１．３８２、ｅｅ＝９９．８４％、［α］^３１．４ _Ｄ＝－４６．１５（ＣＨ_３ＯＨ、ｃ＝０．０５２ｇ／１００ｍＬ）を白色固体として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．５１（ｓ，１Ｈ），８．１９（ｄ，Ｊ＝８．８ Ｈｚ，１Ｈ），７．８９（ｄｄ，Ｊ＝２．１，８．７ Ｈｚ，１Ｈ），７．８２－７．７７（ｍ，２Ｈ），７．４２（ｓ，１Ｈ），６．８２（ｄ，Ｊ＝８．８ Ｈｚ，１Ｈ），５．８８（ｔ，Ｊ＝１１．１ Ｈｚ，１Ｈ），４．４６－４．３３（ｍ，１Ｈ），３．５６－３．３７（ｍ，２Ｈ），２．７１（ｄ，Ｊ＝５．１ Ｈｚ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４３５．１，４３７．１［Ｍ＋Ｈ］^＋． I-15 and I-16 (isomers of I-27)

Step 1: 3-[(5S)-3-chloro-4,5-dihydroisoxazol-5-yl]-N-methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]benzene Sulfonamide

3-(3-bromo-4,5-dihydroisoxazol-5-yl)-N-methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]benzenesulfonamide (900 mg, 723. Hydrochloric acid (2 mL) was added to a solution of 35 μmol, purity 38.5%, 1 equivalent) in 1,4-dioxane (10 mL). The mixture was stirred at 40°C for 12 hours. The reaction mixture was quenched by adding saturated NaHCO ₃ (100 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Agela DuraShell C ₁₈ 150*25 mm*5 μm; Phase: [Water (0.05% _NH3H2O + 10mM _NH4HCO3 )-ACN]; B%: 40%-70%, 10 _min ) and then lyophilized to obtain _the mixture. , this was applied to a chiral SFC column (Column: Column: DAICEL CHIRALPAK AS (250 mm * 30 mm, 10 μm); Mobile phase: [0.1% NH ₃ H ₂ O in EtOH]; B%: 25%-20%) Peak 1 and peak 2 were obtained by separation. Peak 1 was concentrated under reduced pressure to obtain a residue, which was dissolved in MeCN (10 mL) and H ₂ O (20 mL) and then lyophilized to give 3-[(5S)-3-chloro-4,5-dihydro isoxazol-5-yl]-N-methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]benzenesulfonamide (48.36 mg, 111.22 μmol, yield 15.4%, purity 100.0%, SFC: R _t = 1.382, ee = 99.84%, [α] ^31.4 _D = -46.15 (CH ₃ OH, c = 0.052 g/100 mL) as a white solid. ^1H NMR (400 MHz, _CDCl3 ) δ ppm 8.51 (s, 1H), 8.19 (d, J=8.8 Hz, 1H), 7.89 (dd, J=2. 1,8.7 Hz, 1H), 7.82-7.77 (m, 2H), 7.42 (s, 1H), 6.82 (d, J=8.8 Hz, 1H), 5. 88 (t, J=11.1 Hz, 1H), 4.46-4.33 (m, 1H), 3.56-3.37 (m, 2H), 2.71 (d, J=5. 1 Hz, 3H); ES-LCMS m/z 435.1, 437.1 [M+H] ⁺ .

５－ブロモ－６－クロロ－ピリジン－３－スルホニルクロリド（１．０ｇ、３．４４ ｍｍｏｌ、１当量）及びＥｔ_３Ｎ（６９５．５８ｍｇ、６．８７ｍｍｏｌ、９５６．７８μＬ、２当量）の、ＴＨＦ（１０ｍＬ）溶液に、１－（４－メトキシフェニル）－Ｎ－メチル－メタンアミン（５７１．６７ｍｇ、３．７８ｍｍｏｌ、１．１当量）を加えた。混合物を－３０℃で１時間撹拌した。ＴＬＣ（ＰＥ／ＥｔＯＡｃ＝３／１、Ｒ_ｆ＝０．５６）は、出発物質が完全に消費され、新しいスポットが１つ形成されたことを示した。混合物を水（５０ｍＬ）に注ぎ、沈殿した固体を濾過して乾燥させ、５－ブロモ－６－クロロ－Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－ピリジン－３－スルホンアミド（１．１ｇ、２．４９ｍｍｏｌ、収率７２．６％、純度９２．０％）を淡黄色固体として得、これをさらに精製することなく、次工程で使用した。^１Ｈ ＮＭＲ（５００ ＭＨｚ， ＣＤＣｌ_３）δ ｐｐｍ ８．７１（ｄ，Ｊ＝２．１ Ｈｚ，１Ｈ），８．２３（ｄ，Ｊ＝２．１ Ｈｚ，１Ｈ），７．２１（ｄ，Ｊ＝８．７ Ｈｚ，２Ｈ），６．９０－６．８５（ｍ，２Ｈ），４．１８（ｓ，２Ｈ），３．８１（ｓ，３Ｈ），２．７０（ｓ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４０５．０，４０７．０［Ｍ＋Ｈ］^＋． I-17 and I-18 (isomers of I-28)
Step 1: 5-bromo-6-chloro-N-[(4-methoxyphenyl)methyl]-N-methyl-pyridine-3-sulfonamide

5-bromo-6-chloro-pyridine-3-sulfonyl chloride (1.0 g, 3.44 mmol, 1 eq.) and Et ₃ N (695.58 mg, 6.87 mmol, 956.78 μL, 2 eq.) in THF (10 mL) solution was added 1-(4-methoxyphenyl)-N-methyl-methanamine (571.67 mg, 3.78 mmol, 1.1 eq.). The mixture was stirred at -30°C for 1 hour. TLC (PE/EtOAc=3/1, R _f =0.56) showed complete consumption of starting material and formation of one new spot. The mixture was poured into water (50 mL) and the precipitated solid was filtered and dried to give 5-bromo-6-chloro-N-[(4-methoxyphenyl)methyl]-N-methyl-pyridine-3-sulfonamide ( 1.1 g, 2.49 mmol, 72.6% yield, 92.0% purity) was obtained as a pale yellow solid, which was used in the next step without further purification. ^1H NMR (500 MHz, _CDCl3 ) δ ppm 8.71 (d, J=2.1 Hz, 1H), 8.23 (d, J=2.1 Hz, 1H), 7.21 (d, J = 8.7 Hz, 2H), 6.90-6.85 (m, 2H), 4.18 (s, 2H), 3.81 (s, 3H), 2.70 (s, 3H); ES-LCMS m/z 405.0, 407.0 [M+H] ⁺ .

５－ブロモ－６－クロロ－Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－ピリジン－３－スルホンアミド（９５０ｍｇ、２．１５ｍｍｏｌ、純度９２％、１当量）のＴＨＦ（５ｍＬ）溶液に、ＮＨ_３・Ｈ_２Ｏ（１．４８ｍＬ、純度２８％、５当量）を加えた。混合物をマイクロ波の下、１００℃で１２時間撹拌した。ＴＬＣ（ＰＥ／ＥｔＯＡｃ＝１／１、Ｒ_ｆ＝０．３１）は、出発物質が完全に消費され、新しいスポットが１つ形成されたことを示した。溶媒を除去し、６－アミノ－５－ブロモ－Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－ピリジン－３－スルホンアミド（８６０ｍｇ、２．１２ｍｍｏｌ、収率９８．２％、純度９５．０％）を黄色固体として得、これをさらに精製することなく、次工程で使用した。^１Ｈ ＮＭＲ （５００ ＭＨｚ，ＤＭＳＯ－ｄ_６）δ ｐｐｍ ８．３３（ｄ，Ｊ＝２．０ Ｈｚ，１Ｈ），７．９９（ｄ，Ｊ＝２．０ Ｈｚ，１Ｈ），７．３１（ｂｒ ｓ，２Ｈ），７．２３（ｄ，Ｊ＝８．４ Ｈｚ，３Ｈ），６．９２（ｄ，Ｊ＝８．５ Ｈｚ，２Ｈ），３．７５（ｓ，３Ｈ）． Step 2: 6-amino-5-bromo-N-[(4-methoxyphenyl)methyl]-N-methyl-pyridine-3-sulfonamide

A solution of 5-bromo-6-chloro-N-[(4-methoxyphenyl)methyl]-N-methyl-pyridine-3-sulfonamide (950 mg, 2.15 mmol, 92% purity, 1 eq.) in THF (5 mL) To the solution was added NH ₃ .H ₂ O (1.48 mL, 28% purity, 5 eq.). The mixture was stirred at 100° C. for 12 hours under microwave. TLC (PE/EtOAc=1/1, R _f =0.31) showed complete consumption of starting material and formation of one new spot. The solvent was removed and 6-amino-5-bromo-N-[(4-methoxyphenyl)methyl]-N-methyl-pyridine-3-sulfonamide (860 mg, 2.12 mmol, yield 98.2%, purity 95.0%) as a yellow solid, which was used in the next step without further purification. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.33 (d, J = 2.0 Hz, 1H), 7.99 (d, J = 2.0 Hz, 1H), 7.31 ( br s, 2H), 7.23 (d, J=8.4 Hz, 3H), 6.92 (d, J=8.5 Hz, 2H), 3.75 (s, 3H).

６－アミノ－５－ブロモ－Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－ピリジン－３－スルホンアミド（５１０ｍｇ、１．２５ｍｍｏｌ、純度９５％、１当量）及び１－ヨード－４－（トリフルオロメチル）ベンゼン（４０９．４２ｍｇ、１．５１ｍｍｏｌ、２２１．３１μＬ、１．２当量）の、アニソール（２０ｍＬ）溶液に、Ｐｄ（ＯＡｃ）_２（４２．２４ｍｇ、１８８．１５μｍｏｌ、０．１５当量）、キサントホス（７２．５８ｍｇ、１２５．４３μｍｏｌ、０．１当量）、及びＣｓ_２ＣＯ_３（６１３．０２ｍｇ、１．８８ｍｍｏｌ、１．５当量）を加えた。混合物を、Ｎ_２雰囲気下、１３０℃で１６時間撹拌した。溶媒を除去し、残渣をＥｔＯＡｃ（３０ｍＬ）を処理した。混合物を濾過し、濾液を濃縮して残渣を得、これをフラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１／０から３／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝３／１、Ｒｆ＝０．５７）により精製して、５－ブロモ－Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－６－［４－（トリフルオロメチル）アニリノ］ピリジン－３－スルホンアミド（５８７ｍｇ、１．０５ｍｍｏｌ、収率８３．８％、純度９５．０％）を黄色固体として得た。^１Ｈ ＮＭＲ（５００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．６１（ｄ，Ｊ＝１．８ Ｈｚ，１Ｈ），８．１２（ｄ，Ｊ＝１．８ Ｈｚ，１Ｈ），７．８１（ｄ，Ｊ＝８．５Ｈｚ，２Ｈ），７．６４（ｄ，Ｊ＝８．４ Ｈｚ，２Ｈ），７．５１（ｓ，１Ｈ），７．２３（ｄ，Ｊ＝８．５ Ｈｚ，２Ｈ），６．８８（ｄ，Ｊ＝８．５Ｈｚ，２Ｈ），４．１４（ｓ，２Ｈ），３．８１（ｓ，３Ｈ），２．６５（ｓ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ５３０．０，５３２．０［Ｍ＋Ｈ］^＋． Step 3: 5-bromo-N-[(4-methoxyphenyl)methyl]-N-methyl-6-[4-(trifluoromethyl)anilino]pyridine-3-sulfonamide

6-Amino-5-bromo-N-[(4-methoxyphenyl)methyl]-N-methyl-pyridine-3-sulfonamide (510 mg, 1.25 mmol, 95% purity, 1 equivalent) and 1-iodo-4 A solution of -(trifluoromethyl)benzene (409.42 mg, 1.51 mmol, 221.31 μL, 1.2 eq.) in anisole (20 mL) was added Pd(OAc) ₂ (42.24 mg, 188.15 μmol, 0.5 μmol). 15 eq.), xantophos (72.58 mg, 125.43 μmol, 0.1 eq.), and Cs ₂ CO ₃ (613.02 mg, 1.88 mmol, 1.5 eq.) were added. The mixture was stirred at 130 °C for 16 h under _N2 atmosphere. The solvent was removed and the residue was treated with EtOAc (30 mL). The mixture was filtered and the filtrate was concentrated to give a residue, which was purified by flash silica gel chromatography (PE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=3/1, Rf=0.57). Purified to give 5-bromo-N-[(4-methoxyphenyl)methyl]-N-methyl-6-[4-(trifluoromethyl)anilino]pyridine-3-sulfonamide (587 mg, 1.05 mmol, yield 83.8%, purity 95.0%) was obtained as a yellow solid. ^1H NMR (500 MHz, _CDCl3 ) δ ppm 8.61 (d, J=1.8 Hz, 1H), 8.12 (d, J=1.8 Hz, 1H), 7.81 (d, J = 8.5 Hz, 2H), 7.64 (d, J = 8.4 Hz, 2H), 7.51 (s, 1H), 7.23 (d, J = 8.5 Hz, 2H), 6.88 (d, J=8.5Hz, 2H), 4.14 (s, 2H), 3.81 (s, 3H), 2.65 (s, 3H); ES-LCMS m/z 530. 0,532.0 [M+H] ⁺ .

５－ブロモ－Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－６－［４－（トリフルオロメチル）アニリノ］ピリジン－３－スルホンアミド（５８７ｍｇ、１．０５ｍｍｏｌ、純度９５％、１当量）、及び４，４，５，５－テトラメチル－２－ビニル－１，３，２－ジオキサボロラン（３２３．８８ｍｇ、２．１０ｍｍｏｌ、３５６．７０μＬ、２当量）の、１，４－ジオキサン（１８ｍＬ）及びＨ_２Ｏ（３ｍＬ）との溶液に、Ｐｄ（ｄｐｐｆ）Ｃｌ_２（７６．９４ｍｇ、１０５．１５μｍｏｌ、０．１当量）及びＣｓ_２ＣＯ_３（６８５．１７ｍｇ、２．１０ｍｍｏｌ、２当量）を加えた。混合物を、Ｎ_２雰囲気下、９０℃で１６時間撹拌した。溶媒を除去し、残渣をＥｔＯＡｃ（２０ｍＬ）を処理した。混合物を濾過し、濾液を濃縮して残渣を得、これをフラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１／０から３／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝３／１、Ｒ_ｆ＝０．４９）により精製して、Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－６－［４－（トリフルオロメチル）アニリノ］－５－ビニル－ピリジン－３－スルホンアミド（３５０ｍｇ、７３２．９９μｍｏｌ、収率６９．７％、純度１００．０％）を黄色固体として得た。^１Ｈ ＮＭＲ（５００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．６２（ｄ，Ｊ＝２．３ Ｈｚ，１Ｈ），７．８９（ｄ，Ｊ＝２．３ Ｈｚ，１Ｈ），７．７５（ｄ，Ｊ＝８．５ Ｈｚ，２Ｈ），７．６１（ｄ，Ｊ＝８．５ Ｈｚ，２Ｈ），７．２３（ｄ，Ｊ＝８．５ Ｈｚ，２Ｈ），６．８７（ｄ，Ｊ＝８．５ Ｈｚ，３Ｈ），６．７５（ｄｄ，Ｊ＝１１．１，１７．３ Ｈｚ，１Ｈ），５．８４（ｄ，Ｊ＝１７．２ Ｈｚ，１Ｈ），５．７１（ｄ，Ｊ＝１１．１ Ｈｚ，１Ｈ），４．１３（ｓ，２Ｈ），３．８０（ｓ，３Ｈ），２．６３（ｓ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４７８．２［Ｍ＋Ｈ］^＋． Step 4: N-[(4-methoxyphenyl)methyl]-N-methyl-6-[4-(trifluoromethyl)anilino]-5-vinyl-pyridine-3-sulfonamide

5-Bromo-N-[(4-methoxyphenyl)methyl]-N-methyl-6-[4-(trifluoromethyl)anilino]pyridine-3-sulfonamide (587 mg, 1.05 mmol, purity 95%, 1 and 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (323.88 mg, 2.10 mmol, 356.70 μL, 2 eq.) in 1,4-dioxane ( Pd(dppf)Cl ₂ (76.94 mg, 105.15 μmol, 0.1 eq.) and Cs ₂ CO ₃ (685.17 mg, 2.10 mmol, 2 eq.) in a solution with 18 mL) and H ₂ O (3 mL). ) was added. The mixture was stirred at 90 °C for 16 h under _N2 atmosphere. The solvent was removed and the residue was treated with EtOAc (20 mL). The mixture was filtered and the filtrate was concentrated to give a residue, which was subjected to flash silica gel chromatography (PE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=3/1, R _f =0.49). to give N-[(4-methoxyphenyl)methyl]-N-methyl-6-[4-(trifluoromethyl)anilino]-5-vinyl-pyridine-3-sulfonamide (350 mg, 732.99 μmol , yield 69.7%, purity 100.0%) was obtained as a yellow solid. ^1H NMR (500 MHz, _CDCl3 ) δ ppm 8.62 (d, J=2.3 Hz, 1H), 7.89 (d, J=2.3 Hz, 1H), 7.75 (d, J = 8.5 Hz, 2H), 7.61 (d, J = 8.5 Hz, 2H), 7.23 (d, J = 8.5 Hz, 2H), 6.87 (d, J = 8.5 Hz, 3H), 6.75 (dd, J=11.1, 17.3 Hz, 1H), 5.84 (d, J=17.2 Hz, 1H), 5.71 (d, J=11.1 Hz, 1H), 4.13 (s, 2H), 3.80 (s, 3H), 2.63 (s, 3H); ES-LCMS m/z 478.2 [M+H ^] ．．

Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－６－［４－（トリフルオロメチル）アニリノ］－５－ビニル－ピリジン－３－スルホンアミド（３００ｍｇ、６２８．２７μｍｏｌ、純度１００％、１当量）のＤＣＭ（５ｍＬ）溶液に、ＴＦＡ（１．５ｍＬ）を加えた。混合物を２５℃で１６時間撹拌した。ＴＬＣ（ＰＥ／ＥｔＯＡｃ＝１／１、Ｒ_ｆ＝０．５８）は、出発物質が完全に消費され、新しいスポットが１つ形成されたことを示した。混合物を濃縮してＮ－メチル－６－［４－（トリフルオロメチル）アニリノ］－５－ビニル－ピリジン－３－スルホンアミド（３３０ｍｇ、６１６．０７μｍｏｌ、収率９８．１％、純度８８．０％、ＴＦＡ）をオフホワイト固体として得、これをさらに精製することなく、次工程で使用した。^１Ｈ ＮＭＲ （５００ ＭＨｚ，ＣＤ_３ＯＤ）δ ｐｐｍ ８．５０（ｄ，Ｊ＝２．３ Ｈｚ，１Ｈ），８．０９（ｄ，Ｊ＝２．１ Ｈｚ，１Ｈ），７．８５（ｄ，Ｊ＝８．５ Ｈｚ，２Ｈ），７．６２（ｄ，Ｊ＝８．５ Ｈｚ，２Ｈ），７．０５（ｄｄ，Ｊ＝１０．９，１７．２ Ｈｚ，１Ｈ），５．９１（ｄ，Ｊ＝１７．１ Ｈｚ，１Ｈ），５．６２（ｄ，Ｊ＝１１．０Ｈｚ，１Ｈ），２．５９（ｓ，３Ｈ）． Step 5: N-methyl-6-[4-(trifluoromethyl)anilino]-5-vinyl-pyridine-3-sulfonamide

N-[(4-methoxyphenyl)methyl]-N-methyl-6-[4-(trifluoromethyl)anilino]-5-vinyl-pyridine-3-sulfonamide (300 mg, 628.27 μmol, purity 100%, To a solution of 1 eq.) in DCM (5 mL) was added TFA (1.5 mL). The mixture was stirred at 25°C for 16 hours. TLC (PE/EtOAc=1/1, R _f =0.58) showed complete consumption of starting material and formation of one new spot. The mixture was concentrated to give N-methyl-6-[4-(trifluoromethyl)anilino]-5-vinyl-pyridine-3-sulfonamide (330 mg, 616.07 μmol, yield 98.1%, purity 88.0 %, TFA) as an off-white solid, which was used in the next step without further purification. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 8.50 (d, J = 2.3 Hz, 1H), 8.09 (d, J = 2.1 Hz, 1H), 7.85 (d , J=8.5 Hz, 2H), 7.62 (d, J=8.5 Hz, 2H), 7.05 (dd, J=10.9, 17.2 Hz, 1H), 5.91 (d, J=17.1 Hz, 1H), 5.62 (d, J=11.0Hz, 1H), 2.59 (s, 3H).

Ｎ－メチル－６－［４－（トリフルオロメチル）アニリノ］－５－ビニル－ピリジン－３－スルホンアミド（３３０ｍｇ、６１６．０７μｍｏｌ、純度８８％、１当量、ＴＦＡ）及びジブロモメタノンオキシム（２４９．９２ｍｇ、１．２３ｍｍｏｌ、２当量）の、ＥｔＯＡｃ（２０ｍＬ）溶液に、ＮａＨＣＯ_３（５１７．５４ｍｇ、６．１６ｍｍｏｌ、１０当量）を加えた。混合物を２５℃で３時間撹拌した。ＴＬＣ（ＰＥ／ＥｔＯＡｃ＝１／１、Ｒ_ｆ＝０．７８）は、出発物質が完全に消費され、新しいスポットが１つ形成されたことを示した。混合物を濾過し、濾液を濃縮して残渣を得、これをフラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１／０から２／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝１／１、Ｒ_ｆ＝０．７８）により精製して、５－（３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル）－Ｎ－メチル－６－［４－（トリフルオロメチル）アニリノ］ピリジン－３－スルホンアミド（２９０ｍｇ、５７４．８３μｍｏｌ、収率９３．３％、純度９５．０％）を黄色固体として得た。^１Ｈ ＮＭＲ （４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．７１（ｄ，Ｊ＝２．２ Ｈｚ，１Ｈ），７．８５（ｄ，Ｊ＝２．０ Ｈｚ，１Ｈ），７．７４－７．６６（ｍ，２Ｈ），７．６４－７．５７（ｍ，２Ｈ），５．７７（ｔ，Ｊ＝１１．２ Ｈｚ，１Ｈ），４．３９（ｑ，Ｊ＝５．４ Ｈｚ，１Ｈ），４．１２（ｑ，Ｊ＝７．１ Ｈｚ，１Ｈ），３．５６（ｄｄ，Ｊ＝１．８，１１．４ Ｈｚ，２Ｈ），２．７１（ｄ，Ｊ＝５．４ Ｈｚ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４７９．０，４８１．０［Ｍ＋Ｈ］^＋． Step 6: 5-(3-bromo-4,5-dihydroisoxazol-5-yl)-N-methyl-6-[4-(trifluoromethyl)anilino]pyridine-3-sulfonamide

N-Methyl-6-[4-(trifluoromethyl)anilino]-5-vinyl-pyridine-3-sulfonamide (330 mg, 616.07 μmol, 88% purity, 1 eq., TFA) and dibromomethanone oxime (249 To a solution of .92 mg, 1.23 mmol, 2 eq.) in EtOAc (20 mL) was added _NaHCO3 (517.54 mg, 6.16 mmol, 10 eq.). The mixture was stirred at 25°C for 3 hours. TLC (PE/EtOAc=1/1, R _f =0.78) showed complete consumption of starting material and formation of one new spot. The mixture was filtered and the filtrate was concentrated to give a residue, which was subjected to flash silica gel chromatography (PE/EtOAc=1/0 to 2/1, TLC: PE/EtOAc=1/1, R _f =0.78). to give 5-(3-bromo-4,5-dihydroisoxazol-5-yl)-N-methyl-6-[4-(trifluoromethyl)anilino]pyridine-3-sulfonamide (290 mg, 574.83 μmol, yield 93.3%, purity 95.0%) was obtained as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.71 (d, J=2.2 Hz, 1H), 7.85 (d, J=2.0 Hz, 1H), 7.74-7. 66 (m, 2H), 7.64-7.57 (m, 2H), 5.77 (t, J = 11.2 Hz, 1H), 4.39 (q, J = 5.4 Hz, 1H ), 4.12 (q, J = 7.1 Hz, 1H), 3.56 (dd, J = 1.8, 11.4 Hz, 2H), 2.71 (d, J = 5.4 Hz) , 3H); ES-LCMS m/z 479.0, 481.0 [M+H] ⁺ .

５－（３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル）－Ｎ－メチル－６－［４－（トリフルオロメチル）アニリノ］ピリジン－３－スルホンアミド（１００ｍｇ、１９８．２２μｍｏｌ、純度９５％、１当量）をキラルＳＦＣ（カラム：ＤＡＩＣＥＬ ＣＨＩＲＡＬＰＡＫ ＡＤ（２５０ｍｍ＊３０ｍｍ、１０ｕｍ）；移動相：［０．１％のＮＨ_３Ｈ_２Ｏ ＥｔＯＨ］；Ｂ％：３０％－３０％）により分離し、ピーク１及びピーク２を得た。ピーク１を減圧濃縮して残渣を得、これをＭｅＣＮ（１０ｍＬ）及びＨ_２Ｏ（２０ｍＬ）に溶解させ、（Ｓ）－５－（３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル）－Ｎ－メチル－６－（（４－（トリフルオロメチル）フェニル）アミノ）ピリジン－３－スルホンアミド（３２．７９ｍｇ、６８．４２μｍｏｌ、収率３４．５％、純度１００．０％、ＳＦＣ：Ｒ_ｔ＝１．３７２、ｅｅ＝１００％、［α］^２８．６ _Ｄ＝－５．００（ＭｅＯＨ、ｃ＝０．０８ｇ／１００ｍＬ）を白色固体として得た。^１Ｈ ＮＭＲ（５００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．７１（ｄ，Ｊ＝２．１ Ｈｚ，１Ｈ），７．８５（ｄ，Ｊ＝２．１ Ｈｚ，１Ｈ），７．７２－７．６５（ｍ，３Ｈ），７．６４－７．５８（ｍ，２Ｈ），５．７７（ｔ，Ｊ＝１１．２ Ｈｚ，１Ｈ），４．４４（ｑ，Ｊ＝５．２ Ｈｚ，１Ｈ），３．６２－３．５１（ｍ，２Ｈ），２．７１（ｄ，Ｊ＝５．３ Ｈｚ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４７８．９，４８０．９［Ｍ＋Ｈ］^＋．また、ピーク２を減圧濃縮して残渣を得、これをＭｅＣＮ（１０ｍＬ）及びＨ_２Ｏ（２０ｍＬ）に溶解させて凍結乾燥させ、（Ｒ）－５－（３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル）－Ｎ－メチル－６－（（４－（トリフルオロメチル）フェニル）アミノ）ピリジン－３－スルホンアミド（３１．６４ｍｇ、６６．０２μｍｏｌ、収率３３．３％、純度１００．０％、ＳＦＣ：Ｒ_ｔ＝１．５９６、ｅｅ＝１００％、［α］^２８．８ _Ｄ＝＋６．６７（ＭｅＯＨ、ｃ＝０．０９ｇ／１００ｍＬ）を白色固体として得た。^１Ｈ ＮＭＲ （５００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．７０（ｄ，Ｊ＝２．３ Ｈｚ，１Ｈ），７．８６（ｄ，Ｊ＝２．１ Ｈｚ，１Ｈ），７．７２－７．６５（ｍ，３Ｈ），７．６３－７．５９（ｍ，２Ｈ），５．７７（ｔ，Ｊ＝１１．１ Ｈｚ，１Ｈ），４．４８（ｑ，Ｊ＝５．３ Ｈｚ，１Ｈ），３．６２－３．５１（ｍ，２Ｈ），２．７０（ｄ，Ｊ＝５．３ Ｈｚ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４７９．０，４８１．０［Ｍ＋Ｈ］^＋． Step 7: (S)-5-(3-bromo-4,5-dihydroisoxazol-5-yl)-N-methyl-6-((4-(trifluoromethyl)phenyl)amino)pyridine-3- Sulfonamide

5-(3-bromo-4,5-dihydroisoxazol-5-yl)-N-methyl-6-[4-(trifluoromethyl)anilino]pyridine-3-sulfonamide (100 mg, 198.22 μmol, purity 95%, 1 eq.) by chiral SFC (Column: DAICEL CHIRALPAK AD (250mm*30mm, 10um); Mobile phase: [0.1% _{NH3H2O EtOH} ]; B%: 30%-30%). Separation yielded peak 1 and peak 2. Peak 1 was concentrated under reduced pressure to give a residue, which was dissolved in MeCN (10 mL) and H ₂ O (20 mL) to give (S)-5-(3-bromo-4,5-dihydroisoxazol-5-yl). )-N-methyl-6-((4-(trifluoromethyl)phenyl)amino)pyridine-3-sulfonamide (32.79 mg, 68.42 μmol, yield 34.5%, purity 100.0%, SFC : R _t = 1.372, ee = 100%, [α] ^28.6 _D = -5.00 (MeOH, c = 0.08 g/100 mL) was obtained as a white solid. ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 8.71 (d, J = 2.1 Hz, 1H), 7.85 (d, J = 2.1 Hz, 1H), 7.72-7.65 (m, 3H), 7.64-7.58 (m, 2H), 5.77 (t, J=11.2 Hz, 1H), 4.44 (q, J=5.2 Hz, 1H), 3.62-3 .51 (m, 2H), 2.71 (d, J = 5.3 Hz, 3H); ES-LCMS m/z 478.9, 480.9 [M+H] ⁺ .In addition, peak 2 was concentrated under reduced pressure. to obtain a residue, which was dissolved in MeCN (10 mL) and H ₂ O (20 mL) and lyophilized to give (R)-5-(3-bromo-4,5-dihydroisoxazol-5-yl)- N-Methyl-6-((4-(trifluoromethyl)phenyl)amino)pyridine-3-sulfonamide (31.64 mg, 66.02 μmol, yield 33.3%, purity 100.0%, SFC:R _t = 1.596, ee = 100%, [α] ^28.8 _D = +6.67 (MeOH, c = 0.09 g/100 mL) was obtained as a white solid. ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 8.70 (d, J=2.3 Hz, 1H), 7.86 (d, J=2.1 Hz, 1H), 7.72-7.65 (m, 3H), 7.63 -7.59 (m, 2H), 5.77 (t, J = 11.1 Hz, 1H), 4.48 (q, J = 5.3 Hz, 1H), 3.62-3.51 ( m, 2H), 2.70 (d, J=5.3 Hz, 3H); ES-LCMS m/z 479.0, 481.0 [M+H] ⁺ .

４－ブロモ－２－ヨード－アニリン（２ｇ、６．７１ｍｍｏｌ、１当量）及びトリブチル－（１－メチルイミダゾール－４－イル）スタンナン（２．７２ｇ、６．７１ｍｍｏｌ、９１．５％、１当量）の、ＤＭＦ（２０ｍＬ）との混合物に、Ｐｄ（ｄｐｐｆ）Ｃｌ_２（４９１．２２ｍｇ、６７１．３２μｍｏｌ、０．１当量）を加えた。混合物を、Ｎ_２雰囲気下、１３０℃で１２時間撹拌した。混合物を水（１００ｍＬ）で希釈し、ＥｔＯＡｃで抽出した（１００ｍＬ×３）。合わせた有機相をブライン（５０ｍＬ）で洗浄し、無水Ｎａ_２ＳＯ_４で乾燥させて濾過し、減圧濃縮して残渣を得、フラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１００／１から０／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝０／１、Ｒ_ｆ＝０．３０）により精製して、４－ブロモ－２－（１－メチルイミダゾール－４－イル）アニリン（１ｇ、３．２５ｍｍｏｌ、収率４８．５％、純度８２．０％）を黄色油として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ７．４７－７．４２（ｍ，２Ｈ），７．１３－７．０６（ｍ，２Ｈ），６．５８（ｄ，Ｊ＝８．６ Ｈｚ，１Ｈ），５．７２－５．３４（ｍ，２Ｈ），３．７２（ｓ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ２５２．０，２５４．０［Ｍ＋Ｈ］^＋． I-19 and I-20 (isomers of I-29)
Step 1: 4-bromo-2-(1-methylimidazol-4-yl)aniline

4-Bromo-2-iodo-aniline (2 g, 6.71 mmol, 1 eq.) and tributyl-(1-methylimidazol-4-yl)stannane (2.72 g, 6.71 mmol, 91.5%, 1 eq.) Pd(dppf)Cl 2 (491.22 mg, 671.32 μmol, 0.1 eq.) was added to a mixture of Pd(dppf)Cl ₂ (491.22 mg, 671.32 μmol, 0.1 eq.) with DMF (20 mL). The mixture was stirred at 130 °C for 12 h under _N2 atmosphere. The mixture was diluted with water (100 mL) and extracted with EtOAc (100 mL x 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous Na ₂ SO ₄ , filtered, concentrated under reduced pressure to give a residue, and subjected to flash silica gel chromatography (PE/EtOAc = 100/1 to 0/1, TLC: PE/EtOAc=0/1, R _f =0.30) to give 4-bromo-2-(1-methylimidazol-4-yl)aniline (1 g, 3.25 mmol, yield 48. 5%, purity 82.0%) was obtained as a yellow oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.47-7.42 (m, 2H), 7.13-7.06 (m, 2H), 6.58 (d, J = 8.6 Hz , 1H), 5.72-5.34 (m, 2H), 3.72 (s, 3H); ES-LCMS m/z 252.0, 254.0 [M+H] ⁺ .

４－ブロモ－２－（１－メチルイミダゾール－４－イル）アニリン（１ｇ、３．２５ｍｍｏｌ、８２．０％、１当量）及び４，４，５，５－テトラメチル－２－ビニル－１，３，２－ジオキサボロラン（１．００ｇ、６．５１ｍｍｏｌ、１．１０ｍＬ、２当量）の、１，４－ジオキサン（３０ｍＬ）及びＨ_２Ｏ（６ｍＬ）に、Ｐｄ（ｄｐｐｆ）Ｃｌ_２（２３７．９９ｍｇ、３２５．２５μｍｏｌ、０．１当量）及びＣｓ_２ＣＯ_３（３．１８ｇ、９．７６ｍｍｏｌ、３当量）を加えた。混合物を、Ｎ_２雰囲気下、１００℃で２時間撹拌した。混合物を水（１００ｍＬ）で希釈し、ＥｔＯＡｃで抽出した（１００ｍＬ×３）。合わせた有機相をブライン（５０ｍＬ）で洗浄し、無水Ｎａ_２ＳＯ_４で乾燥させて濾過し、減圧濃縮して残渣を得、フラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１００／１から０／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝０／１、Ｒ_ｆ＝０．３５）により精製して、２－（１－メチルイミダゾール－４－イル）－４－ビニル－アニリン（５００ｍｇ、２．１６ｍｍｏｌ、収率６６．４％、純度８６．０％）を褐色油として得た。^１Ｈ ＮＭＲ （４００ ＭＨｚ，ＤＭＳＯ－ｄ_６）δ ｐｐｍ ７．６８（ｓ，１Ｈ），７．５６（ｄ，Ｊ＝１．２ Ｈｚ，１Ｈ），７．４６（ｄ，Ｊ＝２．０ Ｈｚ，１Ｈ），７．０６（ｄｄ，Ｊ＝２．０，８．２ Ｈｚ，１Ｈ），６．６２（ｄ，Ｊ＝８．２ Ｈｚ，１Ｈ），６．５５（ｄｄ，Ｊ＝１１．０，１７．６ Ｈｚ，１Ｈ），６．４３（ｓ，２Ｈ），５．５３（ｄｄ，Ｊ＝１．２，１７．６ Ｈｚ，１Ｈ），４．９４（ｄｄ，Ｊ＝１．０，１０．８ Ｈｚ，１Ｈ），３．７０（ｓ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ２００．３［Ｍ＋Ｈ］^＋． Step 2: 2-(1-methylimidazol-4-yl)-4-vinyl-aniline

4-bromo-2-(1-methylimidazol-4-yl)aniline (1 g, 3.25 mmol, 82.0%, 1 equivalent) and 4,4,5,5-tetramethyl-2-vinyl-1, 3,2-dioxaborolane (1.00 g, 6.51 mmol, 1.10 mL, 2 eq.) in 1,4-dioxane (30 mL) and H ₂ O (6 mL) was dissolved in Pd(dppf)Cl ₂ (237.99 mg). , 325.25 μmol, 0.1 eq) and Cs ₂ CO ₃ (3.18 g, 9.76 mmol, 3 eq) were added. The mixture was stirred at 100 °C for 2 h under _N2 atmosphere. The mixture was diluted with water (100 mL) and extracted with EtOAc (100 mL x 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous Na ₂ SO ₄ , filtered, concentrated under reduced pressure to give a residue, and subjected to flash silica gel chromatography (PE/EtOAc = 100/1 to 0/1, TLC: PE/EtOAc=0/1, R _f =0.35) to give 2-(1-methylimidazol-4-yl)-4-vinyl-aniline (500 mg, 2.16 mmol, yield 66 .4%, purity 86.0%) was obtained as a brown oil. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 7.68 (s, 1H), 7.56 (d, J = 1.2 Hz, 1H), 7.46 (d, J = 2.0 Hz, 1H), 7.06 (dd, J=2.0, 8.2 Hz, 1H), 6.62 (d, J=8.2 Hz, 1H), 6.55 (dd, J=11 .0, 17.6 Hz, 1H), 6.43 (s, 2H), 5.53 (dd, J=1.2, 17.6 Hz, 1H), 4.94 (dd, J=1. 0, 10.8 Hz, 1H), 3.70 (s, 3H); ES-LCMS m/z 200.3 [M+H] ⁺ .

２－（１－メチルイミダゾール－４－イル）－４－ビニル－アニリン（５００ｍｇ、２．１６ｍｍｏｌ、８６．０％、１当量）のＥｔＯＡｃ（１０ｍＬ）溶液に、ＮａＨＣＯ_３（１．８１ｇ、２１．５８ｍｍｏｌ、８３９．３５μＬ、１０当量）及びジブロモメタノンオキシム（６５６．５９ｍｇ、３．２４ｍｍｏｌ、１．５当量）を加えた。混合物を２５℃で１２時間撹拌した。反応混合物を濾過し、減圧濃縮して残渣を得、これを分取ＴＬＣ（ＰＥ／ＥｔＯＡｃ＝０／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝０／１、Ｒ_ｆ＝０．２０）により精製して、４－（３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル）－２－（１メチルイミダゾール－４－イル）アニリン（１５０ｍｇ、４３４．３５μｍｏｌ、収率２０．１％、純度９３．０％）を黄色油として得た。^１Ｈ ＮＭＲ （５００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ７．４７（ｓ，１Ｈ），７．３４（ｄ，Ｊ＝１．８ Ｈｚ，１Ｈ），７．１７（ｓ，１Ｈ），７．０１－６．９９（ｍ，１Ｈ），６．７１（ｄ，Ｊ＝８．２ Ｈｚ，１Ｈ），５．５７（ｔ，Ｊ＝１０．２ Ｈｚ，２Ｈ），３．８０－３．７１（ｍ，３Ｈ），３．５１（ｄｄ，Ｊ＝１０．８，１７．３ Ｈｚ，１Ｈ），３．２３（ｄｄ，Ｊ＝９．８，１７．３ Ｈｚ，１Ｈ），２．９８－１．９９（ｍ，１Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ３２１．１，３２３．１［Ｍ＋Ｈ］^＋． Step 3: 4-(3-bromo-4,5-dihydroisoxazol-5-yl)-2-(1methylimidazol-4-yl)aniline

To a solution of 2-(1-methylimidazol-4-yl)-4-vinyl-aniline (500 mg, 2.16 mmol, 86.0%, 1 eq.) in EtOAc (10 mL) was added NaHCO ₃ (1.81 g, 21. 58 mmol, 839.35 μL, 10 eq.) and dibromomethanone oxime (656.59 mg, 3.24 mmol, 1.5 eq.) were added. The mixture was stirred at 25°C for 12 hours. The reaction mixture was filtered and concentrated under reduced pressure to give a residue, which was purified by preparative TLC (PE/EtOAc=0/1, TLC:PE/EtOAc=0/1, R _f =0.20). 4-(3-bromo-4,5-dihydroisoxazol-5-yl)-2-(1methylimidazol-4-yl)aniline (150 mg, 434.35 μmol, yield 20.1%, purity 93.0 %) as a yellow oil. ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 7.47 (s, 1H), 7.34 (d, J = 1.8 Hz, 1H), 7.17 (s, 1H), 7.01- 6.99 (m, 1H), 6.71 (d, J = 8.2 Hz, 1H), 5.57 (t, J = 10.2 Hz, 2H), 3.80-3.71 (m , 3H), 3.51 (dd, J=10.8, 17.3 Hz, 1H), 3.23 (dd, J=9.8, 17.3 Hz, 1H), 2.98-1. 99 (m, 1H); ES-LCMS m/z 321.1, 323.1 [M+H] ⁺ .

４－（３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル）－２－（１メチルイミダゾール－４－イル）アニリン（１２０ｍｇ、３４７．４８μｍｏｌ、９３．０％、１当量）のＴＨＦ（５ｍＬ）溶液に、ＤＩＥＡ（１３４．７３ｍｇ、１．０４ｍｍｏｌ、１８１．５７μＬ、３当量）及び１－（ブロモメチル）－４－（トリフルオロメチル）ベンゼン（１６６．１２ｍｇ、６９４．９５μｍｏｌ、１０７．１７μＬ、２当量）を加えた。混合物を２５℃で１２時間撹拌した。混合物を水（３０ｍＬ）で希釈し、ＥｔＯＡｃで抽出した（３０ｍＬ×３）。合わせた有機相をブライン（２０ｍＬ）で洗浄し、無水Ｎａ_２ＳＯ_４で乾燥させて濾過し、減圧濃縮して残渣を得、これを分取ＨＰＬＣ（カラム：Ｂｏｓｔｏｎ Ｐｒｉｍｅ Ｃ１８ １５０＊３０ｍｍ＊５μｍ；移動相：［水（０．０５％のＮＨ_３・Ｈ_２Ｏ＋１０ｍＭのＮＨ_４ＨＣＯ_３）－ＡＣＮ］；Ｂ％：６０％－９０％、１０分）により精製して、凍結乾燥させて生成物を得た。生成物をＳＦＣ（カラム：ＤＡＩＣＥＬ ＣＨＩＲＡＬＰＡＫ ＩＧ （２５０ｍｍ＊５０ｍｍ、１０μｍ）；移動相：［０．１％のＮＨ_３・Ｈ_２Ｏ ＥｔＯＨ］；Ｂ％：６０％－６０％）により分離して、ピーク１及びピーク２を得た。ピーク１を減圧濃縮して残渣を得、これをＭｅＣＮ（２０ｍＬ）及びＨ_２Ｏ（４０ｍＬ）に溶解させ、凍結乾燥させて、４－［（５Ｓ）－３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル］－２－（１－メチルイミダゾール－４－イル）－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］アニリン（２４．６１ｍｇ、５０．７２μｍｏｌ、収率１４．６％、純度９８．８％、ＳＦＣ：Ｒ_ｔ＝２．２４８、ｅｅ＝１００％、［α］^２６．８ _Ｄ＝＋１４０．０００（ＭｅＯＨ、ｃ＝０．１８０ｇ／１００ｍＬ））を白色固体として得た。^１Ｈ ＮＭＲ （５００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．５２（ｓ，１Ｈ），７．５７（ｄ，Ｊ＝８．１ Ｈｚ，２Ｈ），７．４９（ｄ，Ｊ＝１０．２ Ｈｚ，３Ｈ），７．３９（ｄ，Ｊ＝２．０ Ｈｚ，１Ｈ），７．２３（ｓ，１Ｈ），７．００（ｄｄ，Ｊ＝２．１，８．５ Ｈｚ，１Ｈ），６．４９（ｄ，Ｊ＝８．４ Ｈｚ，１Ｈ），５．５７（ｔ，Ｊ＝１０．２ Ｈｚ，１Ｈ），４．５６（ｓ，２Ｈ），３．７７（ｓ，３Ｈ），３．５０（ｄｄ，Ｊ＝１０．７，１７．４ Ｈｚ，１Ｈ），３．２３（ｄｄ，Ｊ＝９．８，１７．３ Ｈｚ，１Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４７８．９，４８０．９［Ｍ＋Ｈ］^＋．ピーク２を減圧濃縮して残渣を得、これをＭｅＣＮ（２０ｍＬ）及びＨ_２Ｏ（４０ｍＬ）に溶解させ、凍結乾燥させ、４－［（５Ｒ）－３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル］－２－（１－メチルイミダゾール－４－イル）－Ｎ－［［４－（トリフルオロメチル）フェニル］メチル］アニリン（２４．５８ｍｇ、５０．６１μｍｏｌ、収率１４．６％、純度９８．７％、ＳＦＣ：Ｒ_ｔ＝３．３０１、ｅｅ＝１００％、［α］^２６．８ _Ｄ＝－１９８．７１（ＭｅＯＨ、ｃ＝０．１５５ｇ／１００ｍＬ））を白色固体として得た。^１Ｈ ＮＭＲ （５００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．５４（ｓ，１Ｈ），７．５８－７．５５（ｍ，２Ｈ），７．４９（ｄ，Ｊ＝９．２ Ｈｚ，３Ｈ），７．３９（ｄ，Ｊ＝２．０ Ｈｚ，１Ｈ），７．２３（ｓ，１Ｈ），７．００（ｄｄ，Ｊ＝２．０，８．４ Ｈｚ，１Ｈ），６．４９（ｄ，Ｊ＝８．５ Ｈｚ，１Ｈ），５．５７（ｔ，Ｊ＝１０．３ Ｈｚ，１Ｈ），４．５６（ｓ，２Ｈ），３．７７（ｓ，３Ｈ），３．５０（ｄｄ，Ｊ＝１０．８，１７．３ Ｈｚ，１Ｈ），３．２３（ｄｄ，Ｊ＝９．８，１７．３ Ｈｚ，１Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４７８．９，４８０．９［Ｍ＋Ｈ］^＋． Step 4: 4-[(5S)-3-bromo-4,5-dihydroisoxazol-5-yl]-2-(1-methylimidazol-4-yl)-N-[[4-(trifluoromethyl ) phenyl]methyl]aniline, and 4-[(5R)-3-bromo-4,5-dihydroisoxazol-5-yl]-2-(1-methylimidazol-4-yl)-N-[[ 4-(trifluoromethyl)phenyl]methyl]aniline

4-(3-Bromo-4,5-dihydroisoxazol-5-yl)-2-(1methylimidazol-4-yl)aniline (120 mg, 347.48 μmol, 93.0%, 1 eq.) in THF ( 5 mL) solution, DIEA (134.73 mg, 1.04 mmol, 181.57 μL, 3 eq.) and 1-(bromomethyl)-4-(trifluoromethyl)benzene (166.12 mg, 694.95 μmol, 107.17 μL, 2 equivalents) were added. The mixture was stirred at 25°C for 12 hours. The mixture was diluted with water (30 mL) and extracted with EtOAc (3 x 30 mL). The combined organic phases were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 μm; Mobile phase: [Water (0.05% _NH3.H2O +10mM _{NH4HCO3 )} -ACN]; B%: 60%-90%, 10 min) and lyophilized to give _the product. I got it. The products were separated by SFC (column: DAICEL CHIRALPAK IG (250mm*50mm, 10μm); mobile phase: [0.1% _{NH3.H2O EtOH} ]; B%: 60%-60%), Peak 1 and peak 2 were obtained. Peak 1 was concentrated in vacuo to give a residue, which was dissolved in MeCN (20 mL) and H ₂ O (40 mL) and lyophilized to give 4-[(5S)-3-bromo-4,5-dihydroiso oxazol-5-yl]-2-(1-methylimidazol-4-yl)-N-[[4-(trifluoromethyl)phenyl]methyl]aniline (24.61 mg, 50.72 μmol, yield 14.6 %, purity 98.8%, SFC: R _t = 2.248, ee = 100%, [α] ^26.8 _D = +140.000 (MeOH, c = 0.180 g/100 mL)) was obtained as a white solid. Ta. ^1H NMR (500 MHz, _CDCl3 ) δ ppm 8.52 (s, 1H), 7.57 (d, J = 8.1 Hz, 2H), 7.49 (d, J = 10.2 Hz, 3H), 7.39 (d, J=2.0 Hz, 1H), 7.23 (s, 1H), 7.00 (dd, J=2.1, 8.5 Hz, 1H), 6. 49 (d, J=8.4 Hz, 1H), 5.57 (t, J=10.2 Hz, 1H), 4.56 (s, 2H), 3.77 (s, 3H), 3. 50 (dd, J = 10.7, 17.4 Hz, 1H), 3.23 (dd, J = 9.8, 17.3 Hz, 1H); ES-LCMS m/z 478.9, 480. 9 [M+H] ⁺ . Peak 2 was concentrated in vacuo to give a residue, which was dissolved in MeCN (20 mL) and H ₂ O (40 mL) and lyophilized to give 4-[(5R)-3-bromo-4,5-dihydroisoxazole. -5-yl]-2-(1-methylimidazol-4-yl)-N-[[4-(trifluoromethyl)phenyl]methyl]aniline (24.58 mg, 50.61 μmol, yield 14.6% , purity 98.7%, SFC: R _t = 3.301, ee = 100%, [α] ^26.8 _D = -198.71 (MeOH, c = 0.155 g/100 mL)) was obtained as a white solid. Ta. ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 8.54 (s, 1H), 7.58-7.55 (m, 2H), 7.49 (d, J=9.2 Hz, 3H), 7.39 (d, J = 2.0 Hz, 1H), 7.23 (s, 1H), 7.00 (dd, J = 2.0, 8.4 Hz, 1H), 6.49 (d , J = 8.5 Hz, 1H), 5.57 (t, J = 10.3 Hz, 1H), 4.56 (s, 2H), 3.77 (s, 3H), 3.50 (dd , J=10.8, 17.3 Hz, 1H), 3.23 (dd, J=9.8, 17.3 Hz, 1H); ES-LCMS m/z 478.9, 480.9 [M+H ] ^＋ ．

５－（３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル）－Ｎ－メチル－６－［４－（トリフルオロメチル）アニリノ］ピリジン－３－スルホンアミド（１８５ｍｇ、３６６．７０μｍｏｌ、純度９５％、１当量）の、１，４－ジオキサン（１０ｍＬ）溶液に、ＨＣｌ（４Ｍ、０．５ｍＬ）を加えた。混合物を４０℃で１６時間撹拌した。溶媒を除去し、残渣を水（１０ｍＬ）で処理して、飽和ＮａＨＣＯ_３水溶液でｐＨ８に調節し、ＥｔＯＡｃで抽出した（２０ｍＬ×２）。合わせた有機層をブライン（２０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、分取ＴＬＣ（ＰＥ／ＥｔＯＡｃ＝１／１、Ｒ_ｆ＝０．７１）により精製して生成物を得、これをキラルＳＦＣ（カラム：ＤＡＩＣＥＬ ＣＨＩＲＡＬＰＡＫ ＡＤ（２５０ｍｍ＊３０ｍｍ、１０ｕｍ）；移動相：［０．１％ＮＨ_３Ｈ_２Ｏ ＥｔＯＨ］；Ｂ％：３５％－３５％）により分離して、ピーク１及びピーク２を得た。ピーク１を減圧濃縮して残渣を得、これをＭｅＣＮ（１０ｍＬ）及びＨ_２Ｏ（２０ｍＬ）に溶解させ、凍結乾燥させ、（Ｓ）－５－（３－クロロ－４，５－ジヒドロイソオキサゾール－５－イル）－Ｎ－メチル－６－（（４－（トリフルオロメチル）フェニル）アミノ）ピリジン－３－スルホンアミド（４３．６５ｍｇ、９７．０９μｍｏｌ、収率２６．５％、純度９６．７％、ＳＦＣ：Ｒ_ｔ＝１．２７９、ｅｅ＝９９．４％、［α］^２４．４ _Ｄ＝－２４．２４（ＭｅＯＨ、ｃ＝０．０８２５ｇ／１００ｍＬ）を白色固体として得た。^１Ｈ ＮＭＲ （５００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．７１（ｄ，Ｊ＝２．４ Ｈｚ，１Ｈ），７．８５（ｄ，Ｊ＝２．３ Ｈｚ，１Ｈ），７．７０－７．６８（ｍ，３Ｈ），７．６４－７．５８（ｍ，２Ｈ），５．８４（ｔ，Ｊ＝１１．２ Ｈｚ，１Ｈ），４．４１（ｂｒ ｓ，１Ｈ），３．６２－３．４１（ｍ，２Ｈ），２．７１（ｄ，Ｊ＝５．３Ｈｚ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４３５．０［Ｍ＋Ｈ］^＋． I-21 and I-22 (isomers of I-30)
Step 1: (S)-5-(3-chloro-4,5-dihydroisoxazol-5-yl)-N-methyl-6-((4-(trifluoromethyl)phenyl)amino)pyridine-3- Sulfonamide

5-(3-bromo-4,5-dihydroisoxazol-5-yl)-N-methyl-6-[4-(trifluoromethyl)anilino]pyridine-3-sulfonamide (185 mg, 366.70 μmol, purity To a solution of 95% (1 eq.) in 1,4-dioxane (10 mL) was added HCl (4M, 0.5 mL). The mixture was stirred at 40°C for 16 hours. The solvent was removed and the residue was treated with water (10 mL), adjusted to pH 8 with saturated aqueous NaHCO ₃ and extracted with EtOAc (2×20 mL). The combined organic layers were washed with brine (20 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give a residue, which was subjected to preparative TLC (PE/EtOAc=1/1, R _f =0.71). The product was purified by chiral SFC (column: DAICEL CHIRALPAK AD (250mm*30mm, 10um); mobile phase: [0.1% NH ₃ H ₂ O EtOH]; B%: 35%-35 %) to obtain peak 1 and peak 2. Peak 1 was concentrated in vacuo to give a residue, which was dissolved in MeCN (10 mL) and H ₂ O (20 mL) and lyophilized to give (S)-5-(3-chloro-4,5-dihydroisoxazole). -5-yl)-N-methyl-6-((4-(trifluoromethyl)phenyl)amino)pyridine-3-sulfonamide (43.65 mg, 97.09 μmol, yield 26.5%, purity 96. 7%, SFC: R _t = 1.279, ee = 99.4%, [α] ^24.4 _D = -24.24 (MeOH, c = 0.0825 g/100 mL) was obtained as a white solid. ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 8.71 (d, J = 2.4 Hz, 1H), 7.85 (d, J = 2.3 Hz, 1H), 7.70-7.68 (m, 3H), 7.64-7.58 (m, 2H), 5.84 (t, J=11.2 Hz, 1H), 4.41 (br s, 1H), 3.62-3 .41 (m, 2H), 2.71 (d, J=5.3Hz, 3H); ES-LCMS m/z 435.0 [M+H] ⁺ .

５－（トリフルオロメチル）ピリジン－２－アミン（９１．１１ｍｇ、５６２．００μｍｏｌ、１．５当量）のＤＭＦ（３ｍＬ）溶液に、ＮａＨ（５９．９４ｍｇ、１．５０ｍｍｏｌ、純度６０％、４当量）を加え、混合物を０℃で０．５時間撹拌した。５－ブロモ－６－クロロ－Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－ピリジン－３－スルホンアミド（１６０ｍｇ、３７４．６７μｍｏｌ、純度９５％、１当量）を加え、混合物を２５℃で３時間撹拌した。反応混合物をＨ_２Ｏ（２０ｍＬ）で希釈し、ＥｔＯＡｃで抽出した（４０ｍＬ×３）。合わせた有機相をブライン（２０ｍＬ）で洗浄し、無水Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得た。残渣にＭｅＯＨ（５ｍＬ）を加え、混合物を２５℃で２時間撹拌した。スラリーを濾過し、ケークをＭｅＯＨ（３ｍＬ×２）で洗い流した。固体を収集し、真空乾燥させて、５－ブロモ－Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－６－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］ピリジン－３－スルホンアミド（１６０ｍｇ、３０１．１２μｍｏｌ、収率８０．８％、純度１００．０％）を白色固体として得た。^１Ｈ ＮＭＲ （４００ ＭＨｚ，ＤＭＳＯ－ｄ_６）δ ｐｐｍ ９．２１（ｓ，１Ｈ），８．７２（ｓ，１Ｈ），８．６８（ｄ，Ｊ＝２．２ Ｈｚ，１Ｈ），８．４１（ｄ，Ｊ＝２．２ Ｈｚ，１Ｈ），８．２８－８．１９（ｍ，２Ｈ），７．２４（ｄ，Ｊ＝８．６ Ｈｚ，２Ｈ），６．９３（ｄ，Ｊ＝８．６ Ｈｚ，２Ｈ），４．１６（ｓ，２Ｈ），３．７４（ｓ，３Ｈ），２．６０（ｓ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ５３３．０［Ｍ＋Ｈ］^＋． I-23 and I-24 (isomers of I-31)
Step 1: 5-bromo-N-[(4-methoxyphenyl)methyl]-N-methyl-6-[[5-(trifluoromethyl)-2-pyridyl]amino]pyridine-3-sulfonamide

To a solution of 5-(trifluoromethyl)pyridin-2-amine (91.11 mg, 562.00 μmol, 1.5 eq.) in DMF (3 mL) was added NaH (59.94 mg, 1.50 mmol, purity 60%, 4 eq. ) was added and the mixture was stirred at 0° C. for 0.5 h. 5-Bromo-6-chloro-N-[(4-methoxyphenyl)methyl]-N-methyl-pyridine-3-sulfonamide (160 mg, 374.67 μmol, 95% purity, 1 eq.) was added and the mixture was Stirred at ℃ for 3 hours. The reaction mixture was diluted with H ₂ O (20 mL) and extracted with EtOAc (40 mL x 3). The combined organic phases were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo to give a residue. MeOH (5 mL) was added to the residue and the mixture was stirred at 25° C. for 2 hours. The slurry was filtered and the cake was washed with MeOH (3 mL x 2). The solid was collected and dried under vacuum to give 5-bromo-N-[(4-methoxyphenyl)methyl]-N-methyl-6-[[5-(trifluoromethyl)-2-pyridyl]amino]pyridine- 3-sulfonamide (160 mg, 301.12 μmol, yield 80.8%, purity 100.0%) was obtained as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 9.21 (s, 1H), 8.72 (s, 1H), 8.68 (d, J=2.2 Hz, 1H), 8. 41 (d, J = 2.2 Hz, 1H), 8.28-8.19 (m, 2H), 7.24 (d, J = 8.6 Hz, 2H), 6.93 (d, J =8.6 Hz, 2H), 4.16 (s, 2H), 3.74 (s, 3H), 2.60 (s, 3H); ES-LCMS m/z 533.0 [M+H] ⁺ .

５－ブロモ－Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－６－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］ピリジン－３－スルホンアミド（２６０ｍｇ、４８９．３２μｍｏｌ、純度１００％、１当量）の、１，４－ジオキサン（６ｍＬ）及びＨ_２Ｏ（１ｍＬ）の溶液に、４，４，５，５－テトラメチル－２－ビニル－１，３，２－ジオキサボロラン（３０１．４５ｍｇ、１．９６ｍｍｏｌ、３３１．９９μＬ、４当量）、Ｐｄ（ｄｐｐｆ）Ｃｌ_２（３５．８０ｍｇ、４８．９３μｍｏｌ、０．１当量）、及びＣｓ_２ＣＯ_３（３１８．８６ｍｇ、９７８．６４μｍｏｌ、２当量）を加えた。混合物を、Ｎ_２雰囲気下、９０℃で１２時間撹拌した。反応混合物をＨ_２Ｏ（２０ｍＬ）で希釈し、ＥｔＯＡｃで抽出した（４０ｍＬ×３）。合わせた有機相をブライン（２０ｍＬ）で洗浄し、無水Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、これをフラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１／０から５／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝５／１、Ｒ_ｆ＝０．３５）により精製し、Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－６－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］－５－ビニル－ピリジン－３－スルホンアミド（２００ｍｇ、４０９．６２μｍｏｌ、収率８３．７％、純度９８．０％）を黄色固体として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．６９（ｄ，Ｊ＝２．３ Ｈｚ，１Ｈ），８．６４（ｄ，Ｊ＝８．８Ｈｚ，１Ｈ），８．５５（ｓ，１Ｈ），８．０１－７．９１（ｍ，２Ｈ），７．７９（ｓ，１Ｈ），７．２４（ｄ，Ｊ＝８．８ Ｈｚ，２Ｈ），６．９５－６．８６（ｍ，２Ｈ），６．８２（ｄｄ，Ｊ＝１１．０，１７．３ Ｈｚ，１Ｈ），５．８６（ｄ，Ｊ＝１７．３ Ｈｚ，１Ｈ），５．７４（ｄ，Ｊ＝１１．０ Ｈｚ，１Ｈ），４．１６（ｓ，２Ｈ），３．８１（ｓ，３Ｈ），２．６６（ｓ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４７９．６［Ｍ＋Ｈ］^＋． Step 2: N-[(4-methoxyphenyl)methyl]-N-methyl-6-[[5-(trifluoromethyl)-2-pyridyl]amino]-5-vinyl-pyridine-3-sulfonamide

5-bromo-N-[(4-methoxyphenyl)methyl]-N-methyl-6-[[5-(trifluoromethyl)-2-pyridyl]amino]pyridine-3-sulfonamide (260 mg, 489.32 μmol , 100% pure, 1 eq.) in 1,4-dioxane (6 mL) and H ₂ O (1 mL). Dioxaborolane (301.45 mg, 1.96 mmol, 331.99 μL, 4 eq), Pd(dppf) _Cl2 (35.80 mg, 48.93 μmol, 0.1 eq), and _Cs2CO3 ( _318.86 mg, 978 .64 μmol, 2 equivalents) was added. The mixture was stirred at 90 °C for 12 h under _N2 atmosphere. The reaction mixture was diluted with H ₂ O (20 mL) and extracted with EtOAc (40 mL x 3). The combined organic phases were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was subjected to flash silica gel chromatography (PE/EtOAc = 1/0 to 5/1). , TLC: PE/EtOAc=5/1, R _f =0.35) and purified by N-[(4-methoxyphenyl)methyl]-N-methyl-6-[[5-(trifluoromethyl)- 2-pyridyl]amino]-5-vinyl-pyridine-3-sulfonamide (200 mg, 409.62 μmol, yield 83.7%, purity 98.0%) was obtained as a yellow solid. ^1H NMR (400 MHz, _CDCl3 ) δ ppm 8.69 (d, J = 2.3 Hz, 1H), 8.64 (d, J = 8.8Hz, 1H), 8.55 (s, 1H) ), 8.01-7.91 (m, 2H), 7.79 (s, 1H), 7.24 (d, J=8.8 Hz, 2H), 6.95-6.86 (m, 2H), 6.82 (dd, J = 11.0, 17.3 Hz, 1H), 5.86 (d, J = 17.3 Hz, 1H), 5.74 (d, J = 11.0 Hz, 1H), 4.16 (s, 2H), 3.81 (s, 3H), 2.66 (s, 3H); ES-LCMS m/z 479.6 [M+H] ⁺ .

Ｎ－［（４－メトキシフェニル）メチル］－Ｎ－メチル－６－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］－５－ビニル－ピリジン－３－スルホンアミド（２００ｍｇ、４０９．６２μｍｏｌ、純度９８％、１当量）のＤＣＭ（３ｍＬ）溶液に、ＴＦＡ（１．５１ｇ、１３．２４ｍｍｏｌ、９８０．００μＬ、３２．３１当量）を加えた。混合物を２５℃で３時間撹拌した。溶媒を除去し、Ｎ－メチル－６－［［５－（トリフルオロメチル）－２－ピリジル］アミノ］－５－ビニル－ピリジン－３－スルホンアミド（１４０ｍｇ、粗）を黄色固体として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ８．７１（ｄ，Ｊ＝２．０ Ｈｚ，１Ｈ），８．５２（ｓ，１Ｈ），８．３７（ｄ，Ｊ＝８．８ Ｈｚ，１Ｈ），８．２８（ｓ，１Ｈ），８．１９（ｄ，Ｊ＝７．１ Ｈｚ，１Ｈ），６．７５（ｓ，１Ｈ），６．６１（ｓ，１Ｈ），５．９４（ｄ，Ｊ＝１６．９ Ｈｚ，１Ｈ），５．７８（ｄ，Ｊ＝１１．０ Ｈｚ，１Ｈ），３．９９（ｓ，３Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ３５９．２［Ｍ＋Ｈ］^＋． Step 3: N-methyl-6-[[5-(trifluoromethyl)-2-pyridyl]amino]-5-vinyl-pyridine-3-sulfonamide

N-[(4-methoxyphenyl)methyl]-N-methyl-6-[[5-(trifluoromethyl)-2-pyridyl]amino]-5-vinyl-pyridine-3-sulfonamide (200 mg, 409. To a solution of 62 μmol, 98% purity, 1 eq.) in DCM (3 mL) was added TFA (1.51 g, 13.24 mmol, 980.00 μL, 32.31 eq.). The mixture was stirred at 25°C for 3 hours. The solvent was removed to give N-methyl-6-[[5-(trifluoromethyl)-2-pyridyl]amino]-5-vinyl-pyridine-3-sulfonamide (140 mg, crude) as a yellow solid. ^1H NMR (400 MHz, _CDCl3 ) δ ppm 8.71 (d, J=2.0 Hz, 1H), 8.52 (s, 1H), 8.37 (d, J=8.8 Hz, 1H), 8.28 (s, 1H), 8.19 (d, J = 7.1 Hz, 1H), 6.75 (s, 1H), 6.61 (s, 1H), 5.94 ( d, J = 16.9 Hz, 1H), 5.78 (d, J = 11.0 Hz, 1H), 3.99 (s, 3H); ES-LCMS m/z 359.2 [M+H] ⁺ ．．

Step 4: 5-[(5S)-3-bromo-4,5-dihydroisoxazol-5-yl]-N-methyl-6-[[5-(trifluoromethyl)-2-pyridyl]amino]pyridine -3-sulfonamide, and 5-[(5R)-3-bromo-4,5-dihydroisoxazol-5-yl]-N-methyl-6-[[5-(trifluoromethyl)-2-pyridyl ]amino]pyridine-3-sulfonamide
N-Methyl-6-[[5-(trifluoromethyl)-2-pyridyl]amino]-5-vinyl-pyridine-3-sulfonamide (140 mg, 390.69 μmol, 1 eq.) in EtOAc (10 mL). , NaHCO ₃ (328.22 mg, 3.91 mmol, 151.95 μL, 10 eq.), and dibromomethanone oxime (158.49 mg, 781.38 μmol, 2 eq.) were added. The mixture was stirred at 25°C for 6 hours. The mixture was filtered and the filtrate was concentrated to give a residue, which was subjected to flash silica gel chromatography (PE/EtOAc=1/0 to 1/1, TLC: PE/EtOAc=1/1, R _f =0.32). The product was purified by chiral SFC (column: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 um); mobile phase: [0.1% NH ₃ H ₂ O MeOH]; B%: 55%- 55%) to obtain peak 1 and peak 2. Peak 2 was concentrated under reduced pressure to give 5-[(5R)-3-bromo-4,5-dihydroisoxazol-5-yl]-N-methyl-6-[[5-(trifluoromethyl)-2-pyridyl ]Amino]pyridine-3-sulfonamide (39.4 mg, 81.48 μmol, yield 20.9%, purity 99.3%, SFC: R _t = 4.427, ee = 99.9%, [α] ^24.5 _D = +39.22 (MeOH, c = 0.051 g/100 mL) was obtained as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.73 (s, 1H), 8.68-8.55 (m, 2H), 8.14-8.07 (m, 1H), 8.06-7.98 (m, 2H), 7.63 (q, J = 4.8 Hz, 1H), 6.19 (dd, J=7.9, 11.0 Hz, 1H), 3.92 (dd, J=11.0, 17.5 Hz, 1H), 3.43-3 .39 (m, 1H), 2.45 (d, J=4.9 Hz, 3H); ES-LCMS m/z 480.1 [M+H] ⁺ .

４－（トリフルオロメチル）シクロヘキサンアミン（７４８．２４ｍｇ、４．４８ｍｍｏｌ、１．５当量）のＤＭＳＯ（２５ｍＬ）溶液に、３－ブロモ－４－フルオロ－Ｎ－メチル－ベンゼンスルホンアミド（８００ｍｇ、２．９８ｍｍｏｌ、純度１００．０％、１当量）を加えた。反応混合物を、Ｎ_２雰囲気下、１４０℃で３時間撹拌した。反応混合物をＨ_２Ｏ（３０ｍＬ）で希釈し、ＥｔＯＡｃで抽出した（６０ｍＬ×３）。合わせた有機層をブライン（５０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥さ、濾過し減圧濃縮して残渣を得、これをフラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１００／１から３／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝３／１、Ｒ_ｆ＝０．３６）により精製して粗生成物を得、これを、分取ＨＰＬＣ（カラム：Ｂｏｓｔｏｎ Ｇｒｅｅｎ ＯＤＳ １５０＊３０ｍｍ＊５μｍ；移動相：［水（０．０５％のＨＣｌ）－ＡＣＮ］；Ｂ％：５３％－８３％、１０分）によりさらに精製して、３－ブロモ－Ｎ－メチル－４－［［４－（トリフルオロメチル）シクロヘキシル］アミノ］ベンゼンスルホンアミド（２５０ｍｇ、５７１．９２μｍｏｌ、収率１９．２％、純度９５．０％）を、白色固体として得た。^１Ｈ ＮＭＲ（５００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ７．９２（ｄ，Ｊ＝２．１ Ｈｚ，１Ｈ），７．６５（ｄｄ，Ｊ＝２．１，８．７ Ｈｚ，１Ｈ），６．６５（ｄ，Ｊ＝８．７ Ｈｚ，１Ｈ），４．６８（ｓ，１Ｈ），４．２０（ｓ，１Ｈ），３．３８－３．３４（ｍ，１Ｈ），２．７１－２．５８（ｍ，３Ｈ），２．２７（ｄ，Ｊ＝１１．０ Ｈｚ，２Ｈ），２．１５－２．０３（ｍ，３Ｈ），１．５５－１．４７（ｍ，２Ｈ），１．３４－１．２５（ｍ，２Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４１５．２，４１７．２［Ｍ＋Ｈ］^＋． I-33 and I-34 (isomers of I-32)
Step 1: 3-bromo-N-methyl-4-[[4-(trifluoromethyl)cyclohexyl]amino]benzenesulfonamide

To a solution of 4-(trifluoromethyl)cyclohexaneamine (748.24 mg, 4.48 mmol, 1.5 eq.) in DMSO (25 mL) was added 3-bromo-4-fluoro-N-methyl-benzenesulfonamide (800 mg, 2 .98 mmol, purity 100.0%, 1 equivalent) was added. The reaction mixture was stirred at 140 °C for 3 h under _N2 atmosphere. The reaction mixture was diluted with H ₂ O (30 mL) and extracted with EtOAc (60 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give a residue, which was subjected to flash silica gel chromatography (PE/EtOAc = 100/1 to 3/1; TLC: PE/EtOAc=3/1, R _f =0.36) to obtain a crude product, which was purified by preparative HPLC (column: Boston Green ODS 150*30mm*5μm; mobile phase: [water (0.05% HCl)-ACN]; B%: 53%-83%, 10 min) to produce 3-bromo-N-methyl-4-[[4-(trifluoromethyl)cyclohexyl] ]Amino]benzenesulfonamide (250 mg, 571.92 μmol, yield 19.2%, purity 95.0%) was obtained as a white solid. ^1H NMR (500 MHz, _CDCl3 ) δ ppm 7.92 (d, J=2.1 Hz, 1H), 7.65 (dd, J=2.1, 8.7 Hz, 1H), 6. 65 (d, J=8.7 Hz, 1H), 4.68 (s, 1H), 4.20 (s, 1H), 3.38-3.34 (m, 1H), 2.71-2 .58 (m, 3H), 2.27 (d, J=11.0 Hz, 2H), 2.15-2.03 (m, 3H), 1.55-1.47 (m, 2H), 1.34-1.25 (m, 2H); ES-LCMS m/z 415.2, 417.2 [M+H] ⁺ .

３－ブロモ－Ｎ－メチル－４－［［４－（トリフルオロメチル）シクロヘキシル］アミノ］ベンゼンスルホンアミド（２１０ｍｇ、４４１．６４μｍｏｌ、１当量、ＨＣｌ）及び４，４，５，５－テトラメチル－２－ビニル－１，３，２－ジオキサボロラン（１０２．０３ｍｇ、６６２．４５μｍｏｌ、１１２．３６μＬ、１．５当量）の、１，４－ジオキサン（６ｍＬ）及びＨ_２Ｏ（２ｍＬ）の溶液に、Ｐｄ（ｄｐｐｆ）Ｃｌ_２（３２．３１ｍｇ、４４．１６μｍｏｌ、０．１当量）及びＣｓ_２ＣＯ_３（３５９．７３ｍｇ、１．１０ｍｍｏｌ、２．５当量）を加えた。混合物を、Ｎ_２雰囲気下、１００℃で２時間撹拌した。混合物を水（３０ｍＬ）で希釈し、ＥｔＯＡｃで抽出した（３０ｍＬ×３）。有機層をブライン（３０ｍＬ×２）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、これをフラッシュシリカゲルクロマトグラフィーＰＥ／ＥｔＯＡｃ＝１００／１から１／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝１／１、Ｒ_ｆ＝０．６０）により精製して、Ｎ－メチル－４－［［４－（トリフルオロメチル）シクロヘキシル］アミノ］－３－ビニル－ベンゼンスルホンアミド（１６０ｍｇ、４３５．７５μｍｏｌ、収率９８．７％、純度９８．７％）を得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δｐｐｍ ７．７２－７．５６（ｍ，２Ｈ），６．７１－６．５４（ｍ，２Ｈ），５．６７（ｄ，Ｊ＝１７．４ Ｈｚ，１Ｈ），５．４４（ｄ，Ｊ＝１１．０ Ｈｚ，１Ｈ），４．２０－４．１４（ｍ，１Ｈ），４．１２（ｄ，Ｊ＝７．１ Ｈｚ，１Ｈ），３．３５（ｍ，Ｊ＝４．０，７．４，１１．２ Ｈｚ，１Ｈ），２．６４（ｄ，Ｊ＝５．４ Ｈｚ，３Ｈ），２．２６（ｄ，Ｊ＝１２．０ Ｈｚ，２Ｈ），２．０８（ｓ，１Ｈ），２．０４（ｓ，２Ｈ），１．５４－１．４２（ｍ，２Ｈ），１．２６－１．２２（ｍ，２Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ３６３．２［Ｍ＋Ｈ］^＋． Step 2: N-methyl-4-[[4-(trifluoromethyl)cyclohexyl]amino]-3-vinyl-benzenesulfonamide

3-Bromo-N-methyl-4-[[4-(trifluoromethyl)cyclohexyl]amino]benzenesulfonamide (210 mg, 441.64 μmol, 1 eq., HCl) and 4,4,5,5-tetramethyl- 2-vinyl-1,3,2-dioxaborolane (102.03 mg, 662.45 μmol, 112.36 μL, 1.5 eq.) in a solution of 1,4-dioxane (6 mL) and H ₂ O (2 mL). Pd(dppf) _Cl2 (32.31 _mg , 44.16 [mu]mol, 0.1 eq.) and _Cs2CO3 (359.73 mg, 1.10 mmol, 2.5 eq.) were added. The mixture was stirred at 100 °C for 2 h under _N2 atmosphere. The mixture was diluted with water (30 mL) and extracted with EtOAc (3 x 30 mL). The organic layer was washed with brine (30 mL x 2), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was subjected to flash silica gel chromatography PE/EtOAc = 100/1 to 1/1, TLC. : PE/EtOAc=1/1, R _f =0.60) to give N-methyl-4-[[4-(trifluoromethyl)cyclohexyl]amino]-3-vinyl-benzenesulfonamide (160 mg , 435.75 μmol, yield 98.7%, purity 98.7%). ¹ H NMR (400 MHz, CDCl ₃ ) δppm 7.72-7.56 (m, 2H), 6.71-6.54 (m, 2H), 5.67 (d, J = 17.4 Hz, 1H), 5.44 (d, J=11.0 Hz, 1H), 4.20-4.14 (m, 1H), 4.12 (d, J=7.1 Hz, 1H), 3. 35 (m, J = 4.0, 7.4, 11.2 Hz, 1H), 2.64 (d, J = 5.4 Hz, 3H), 2.26 (d, J = 12.0 Hz) , 2H), 2.08 (s, 1H), 2.04 (s, 2H), 1.54-1.42 (m, 2H), 1.26-1.22 (m, 2H); ES- LCMS m/z 363.2 [M+H] ⁺ .

Ｎ－メチル－４－［［４－（トリフルオロメチル）シクロヘキシル］アミノ］－３－ビニル－ベンゼンスルホンアミド（１１０ｍｇ、２９７．４５μｍｏｌ、１当量）の、ＥｔＯＡｃ（５ｍＬ）溶液に、ジブロモメタノンオキシム（９０．５０ｍｇ、４４６．１８μｍｏｌ、１．５当量）及びＮａＨＣＯ_３（２４９．８８ｍｇ、２．９７ｍｍｏｌ、１０当量）を加えた。混合物を４０℃で１２時間撹拌した。ＴＬＣ（ＰＥ／ＥｔＯＡｃ＝１／１、Ｒ_ｆ＝０．７２）は、出発物質が完全に消費され、新しいスポットが検出されたことを示した。混合物を水（５０ｍＬ）で希釈し、ＥｔＯＡｃで抽出した（５０ｍＬ×３）。有機層をブライン（５０ｍＬ×２）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し減圧濃縮して残渣を得、これをフラッシュシリカゲルクロマトグラフィー（ＰＥ／ＥｔＯＡｃ＝１００／１から１／１、ＴＬＣ：ＰＥ／ＥｔＯＡｃ＝１／１、Ｒ_ｆ＝０．６０）により、及び、分取ＳＦＣ（カラム：ＤＡＩＣＥＬ ＣＨＩＲＡＬＰＡＫ ＡＤ（２５０ｍｍ＊３０ｍｍ、１０ｕｍ）；移動相：［０．１％のＮＨ_３・Ｈ_２Ｏ ＥＴＯＨ］；Ｂ％：２５％－２５％）により精製し、ピーク１及びピーク２を得た。ピーク１を減圧濃縮して残渣を得、これをＭｅＣＮ（２０ｍＬ）及びＨ_２Ｏ（２０ｍＬ）に溶解させ、凍結乾燥させ、３－［（５Ｓ）－３－ブロモ－４，５－ジヒドロイソオキサゾール－５－イル］－Ｎ－メチル－４－［［４－（トリフルオロメチル）シクロヘキシル］アミノ］ベンゼンスルホンアミド（６０ｍｇ、１２２．４０μｍｏｌ、収率４１．１％、純度９８．８％、ＳＦＣ：Ｒ_ｔ＝１．４１４、ｅｅ＝１００％、［α］^２４．４ _Ｄ＝－２３．０７（ＭｅＯＨ、ｃ＝０．０２６ｇ／１００ｍＬ））を黄色固体として得た。^１Ｈ ＮＭＲ（４００ ＭＨｚ，ＣＤＣｌ_３）δ ｐｐｍ ７．７１（ｄ，Ｊ＝８．６ Ｈｚ，１Ｈ），７．５５（ｓ，１Ｈ），６．７５（ｄ，Ｊ＝８．８ Ｈｚ，１Ｈ），５．６６（ｔ，Ｊ＝１１．２ Ｈｚ，１Ｈ），４．６０（ｄ，Ｊ＝７．１ Ｈｚ，１Ｈ），４．２２（ｑ，Ｊ＝４．９ Ｈｚ，１Ｈ），３．５５－３．４０（ｍ，２Ｈ），３．３８－３．２９（ｍ，１Ｈ），２．６６－２．６１（ｍ，３Ｈ），２．２４（ｓ，２Ｈ），２．０７（ｄ，Ｊ＝１２．５ Ｈｚ，３Ｈ），１．５３－１．４４（ｍ，２Ｈ），１．２９－１．１７（ｍ，２Ｈ）；ＥＳ－ＬＣＭＳ ｍ／ｚ ４８６．０［Ｍ＋Ｈ］^＋． Step 3: 3-[(5S)-3-bromo-4,5-dihydroisoxazol-5-yl]-N-methyl-4-[[4-(trifluoromethyl)cyclohexyl]amino]benzenesulfonamide

A solution of N-methyl-4-[[4-(trifluoromethyl)cyclohexyl]amino]-3-vinyl-benzenesulfonamide (110 mg, 297.45 μmol, 1 eq.) in EtOAc (5 mL) was added with dibromomethanone oxime. (90.50 mg, 446.18 μmol, 1.5 eq.) and NaHCO ₃ (249.88 mg, 2.97 mmol, 10 eq.) were added. The mixture was stirred at 40°C for 12 hours. TLC (PE/EtOAc=1/1, R _f =0.72) showed that the starting material was completely consumed and new spots were detected. The mixture was diluted with water (50 mL) and extracted with EtOAc (50 mL x 3). The organic layer was washed with brine (50 mL x 2), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was subjected to flash silica gel chromatography (PE/EtOAc = 100/1 to 1/1; TLC: PE/EtOAc=1/1, R _f =0.60) and preparative SFC (column: DAICEL CHIRALPAK AD (250 mm * 30 mm, 10 um); mobile phase: [0.1% _NH3 . H ₂ O ETOH]; B%: 25%-25%) to obtain peak 1 and peak 2. Peak 1 was concentrated in vacuo to give a residue, which was dissolved in MeCN (20 mL) and H ₂ O (20 mL) and lyophilized to give 3-[(5S)-3-bromo-4,5-dihydroisoxazole. -5-yl]-N-methyl-4-[[4-(trifluoromethyl)cyclohexyl]amino]benzenesulfonamide (60 mg, 122.40 μmol, yield 41.1%, purity 98.8%, SFC: R _t =1.414, ee = 100%, [α] ^24.4 _D = -23.07 (MeOH, c = 0.026 g/100 mL)) was obtained as a yellow solid. ^1H NMR (400 MHz, _CDCl3 ) δ ppm 7.71 (d, J=8.6 Hz, 1H), 7.55 (s, 1H), 6.75 (d, J=8.8 Hz, 1H), 5.66 (t, J=11.2 Hz, 1H), 4.60 (d, J=7.1 Hz, 1H), 4.22 (q, J=4.9 Hz, 1H) , 3.55-3.40 (m, 2H), 3.38-3.29 (m, 1H), 2.66-2.61 (m, 3H), 2.24 (s, 2H), 2 .07 (d, J = 12.5 Hz, 3H), 1.53-1.44 (m, 2H), 1.29-1.17 (m, 2H); ES-LCMS m/z 486.0 [M+H] ⁺ .

Example 2. TEAD Inhibition Assay TEAD inhibition can be assayed using the Hippo pathway TEAD reporter-MCF7 cell line (BPS Bioscience, catalog #: 60618).

Background The Hippo pathway controls cell proliferation and cell death. The Hippo pathway is activated by high cell density and cell stress, arresting cell proliferation and inducing apoptosis. The mammalian Hippo pathway includes MST and LATS kinases. When the Hippo pathway is activated, MST kinase phosphorylates LATS kinase, which phosphorylates the transcriptional coactivators YAP and TAZ. Unphosphorylated YAP and TAZ can translocate to the nucleus and interact with TEAD/TEF transcription factors to initiate cell cycle-promoting gene transcription. However, upon phosphorylation, YAP and TAZ are recruited from the nucleus to the cytosol, thereby terminating YAP and TAZ-dependent gene transcription. Dysfunction of the Hippo pathway is frequently detected in human cancers, and downregulation of the Hippo pathway correlates with the aggressive nature of cancer cells and poor prognosis.

Description The TEAD reporter-MCF7 cell line contains the firefly luciferase gene under the control of a TEAD-responsive element that is stably integrated into the human breast cancer cell line, MCF7. Within the cell, unphosphorylated basal YAP/TAZ remains in the nucleus and induces constitutive expression of the luciferase reporter. Cell lines have been tested for inhibiting expression of luciferase reporters by activators of the Hippo pathway.
Applications • Monitor Hippo pathway activity.
- Screen for activators or inhibitors of the Hippo pathway.

Format Each vial contains approximately 1.5X106 cells in 1 mL of 10% DMSO.
storage

Store in liquid nitrogen immediately after receiving.

General culture conditions Medium 1 (BPS Bioscience #60187) + 10 μg/ml insulin (Sigma-Aldrich # I0516): 10% FBS (Invitrogen #26140-079), 1% non-essential amino acids (Hyclone #SH30238.01) , MEM medium (Hyclone #SH30024.0) supplemented with 1 mM sodium pyruvate (Hyclone #SH30239.01), 1% penicillin/streptomycin (Hyclone SV30010.01), +10 μg/mL insulin (Sigma-Aldrich # I0516) 1 )

Growth Medium 1B (BPS Bioscience #79531) + 10 μg/mL Insulin (Sigma-Aldrich # I0516): Medium 1 (BPS Catalog #60187) + 10 μg/mL Insulin (Sigma-Aldrich # I0516), and 400 μg/mL Geneticin ( Invitrogen #11811031).

Cells should be grown at 37° C., 5% CO ₂ using Growth Medium 1B containing 10 μg/mL insulin. Depending on the _NaHCO3 concentration of the basal medium, it may be necessary to adjust the rate of _CO2 in the incubator.

Immediately thaw the frozen cells from liquid nitrogen in a 37°C water bath, transfer to a tube containing 10 mL of Thawing Medium 1 + Insulin (without geneticin), spin down the cells, and place the cells in prewarmed Thawing Medium 1 + Insulin (free of geneticin). It is recommended to resuspend cells (without geneticin), transfer the resuspended cells to a T25 flask, and culture overnight at 37° C. in a CO ₂ incubator. The next day, the medium is replaced with fresh thawing medium 1+insulin (no geneticin) and growth culture is continued at 37° C. in a CO ₂ incubator until cells are ready to divide. At the first passage, switch to growth medium 1B + 10 μg/mL insulin (contains thawing medium 1, insulin, and geneticin). Cells need to be allowed to divide before reaching full confluence.

To passage the cells, wash them with phosphate buffered saline (PBS) and detach the cells from the culture vessel with 0.25% trypsin/EDTA. Add growth medium 1B + 10 μg/mL insulin (containing thawing medium 1, insulin, and geneticin), transfer to a tube, spin down the cells, then resuspend the cells and aliquot the appropriate cell suspension. Seed into new culture containers. Subculture distribution: 1:5 to 1:10 per week.

To freeze the cells, wash them with phosphate buffered saline (PBS) and detach the cells from the culture vessel with trypsin/EDTA. Add growth medium 1B + 10 μg/mL insulin (containing thawing medium 1, insulin, and geneticin), transfer to tube, spin down cells, and resuspend in freezing medium (10% DMSO + 90% FBS). Place it at -80°C overnight, and the next day, place it in liquid nitrogen. Alternatively, the vial can be placed directly into liquid nitrogen.

Functional Validation and Assay Performance The following assay is designed for a 96-well format. To perform assays in different tissue culture formats, cell numbers and reagent volumes need to be adjusted appropriately.
Materials required for cell culture but not supplied: Thawing medium 1 (BPS Bioscience #60187) + 10 μg/mL insulin Growth medium 1B (BPS Bioscience #79531) + 10 μg/mL insulin Bovine pancreas-derived insulin solution (Sigma- Aldrich #: I0516)
Materials needed but not supplied for cell assays: H ₂ O ₂ : activator of the Hippo pathway (activates MST kinase)
- Insulin Assay Medium: Thawing Medium 1 (BPS Cat. #60187) + 10 μg/mL Insulin Bovine Pancreas Derived Insulin Solution (Sigma-Aldrich Cat. #: I0516)
- Okadaic acid (BPS bioscience #27047): activator of the Hippo pathway (activates MST kinase). Prepare a 10 mM stock solution of DMSO.
- 96-well tissue culture plate or 96-well tissue culture-treated white clear bottom assay plate - ONE-Step™ Luciferase Assay System (BPS, Catalog #60690)
・Luminance meter

Mycoplasma Testing Cell lines were screened using the PCR-based VenorGeM Mycoplasma Detection Kit (Sigma-Aldrich) to confirm the absence of Mycoplasma species.
Inhibition of TEAD Reporter Activity by Activators of the Hippo Pathway in TEAD Reporter-MCF7 Cells 1) Harvest TEAD Reporter-MCF7 cells from the culture in growth medium and place them in 45 μL of assay medium with a white transparent bottom. Cells are seeded in 96-well microplates at a density of 35,000 cells per well.
2) Incubate cells at 37°C overnight in a _CO2 incubator.
3) Dilute the stock solution of activator (H ₂ O ₂ or okadaic acid) in assay medium. Add 5 μL of diluted activator to the wells. The final concentration of DMSO in the assay medium is 0.1%.
4) Add 5 μL of assay medium (same concentration of DMSO without activator) to control wells.
5) Add 50 μL of assay medium (containing DMSO) to cell-free control wells (to measure background luminescence).
6) Set each process in at least three ways.
7) Incubate cells at 37° C. for 5-6 hours in a CO ₂ incubator.
8) Perform the luciferase assay using the ONE-Step™ Luciferase Assay System according to the procedure shown: Add 100 μL of ONE-Step™ Luciferase Reagent per well and shake for approximately 15 minutes at room temperature. Measure luminescence using a luminometer.
9) Data Analysis: Subtract the average background luminescence (cell-free control wells) from the luminescence readings of all wells to obtain background-subtracted luminescence.

Certain compounds were tested in the TEAD reporter assay as well as H226 and H28. The data are shown in Table 2 below. A: EC50<0.1uM; B: 0.1uM≦EC50≦0.5uM; C: EC50>0.5uM.

Example 3: Pharmacokinetic studies in mice Formulated compounds are administered intravenously or orally to BALB/c mice by gavage. Typically, blood is drawn at 0.167, 0.5, 1, 2, 4, 6, 12, and 24 hours after administration, processed by centrifugation into plasma, and stored at -80°C until analysis. Internal standards are added to each sample before protein precipitation with acetonitrile or TCA. The precipitate is filtered through a filter plate and the sample is analyzed by LC/MS/MS. A standard curve is prepared in plasma, typically from 1.0 ng/mL to 3000 ng/mL, and processed in the same manner as the samples. Typically, the same analysis was performed on a suitable LC/MS/MS system adapted to an analytical UPLC column, and compounds were analyzed in 30-95% 0.1% formic acid (v/v) in ACN:0. Elute the analytical column with a gradient of .1% formic acid (v/v). Test compound and internal standard mass spectrometry detection is performed on the MRM in positive mode. The pharmacokinetics of each compound is analyzed by Phoenix WinNonlin software (Pharsight, St. Louis, MO) in a non-compartmental analysis.

Example 4. CTGF Data Analysis NU/NU nude female mice are obtained from Charles River Laboratories and injected subcutaneously with NCI-H226 (ATCC) human mesothelioma cells. Once tumors have grown to an average size of 350-400 mm ³ , mice are randomized into each treatment group. Mice bearing NCI-H226 tumors are treated with vehicle (5% DMSO/95% PEG 400) or TEAD inhibitor by oral gavage for a total of 3 doses. Four hours after the third dose, mice are euthanized and tumors are collected for RNA isolation for pharmacokinetic (PD) analysis.

RNA is extracted from tumors using QIAZOL (Qiagen) cell lysis reagent and the tissue is then homogenized for 10 minutes using a TissueLyser II (Qiagen). Once the sample disruption and digestion is complete, chloroform is added to each sample and the homogenate is separated into aqueous and organic phases by centrifugation.

RNA is then isolated from the sample using the KingFisher Flex automated extraction system and the MagMAX mirvana total RNA isolation kit. For RNA extraction, follow the manufacturer's recommended procedures for high-throughput isolation of RNA from tissue samples.

The expression of the YAP/TEAD regulatory gene, CCN2 encoding CTFG (connective tissue growth factor), and the housekeeping gene human glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was determined using TaqMan gene expression Master Mix and Taqman probe. Quantified by qRT-PCR analysis. CTGF and GAPDH cycle threshold (Ct) values are determined for tumor cDNA samples and CTGF expression is normalized to GAPDH as an internal control.

Relative CTGF mRNA expression levels for each treatment group from tumor tissue are normalized to the vehicle control group. For comparisons between vehicle control and TEAD inhibitor treated groups, independent sample t-tests are used for statistical analysis.

Example 5. Anti-Proliferation Assay Individual cell lines are grown in culture according to the supplier's instructions and seeded into 96-well plates at a density that ensures logarithmic growth for 72-96 hours. TEAD inhibitor compounds are administered to cells at a maximum concentration of 10 μ m followed by 10 point 3-fold serial dilutions. After 72-96 hours, proliferation is quantified using Cell TiterGlo™ (Promega, Inc.) and compared to vehicle controls. IC50 and EC50 values are generated using Prism or XLFit curve fitting software.

Example 6. In Vivo Efficacy Study of NCI-H226 In Vivo Inhibition of Tumor Growth 6-8 week old nu/nu nude mice (CRL) were inoculated subcutaneously on the right flank with 5 × ¹⁰ NCI-H226 human mesothelioma tumor cells. do. Monitor tumor growth twice a week using calipers and calculate mean tumor volume (MTV) using the formula V=W2×L/2.

Once MTV reached approximately 150-200 ^mm3 , animals were randomized into treatment groups (n = 8-10/group) and treated with either vehicle (5% DMSO + 95% PEG 400) or TEAD inhibitor. Administer orally (PO) on a once daily (QD) schedule for ~40 days.

Randomization and treatment begin on day 0, and percent tumor growth inhibition is calculated on the last day of the study (the day the control MTV reaches the maximum allowed tumor volume) and the following calculations are performed.

%TGI = 100 - [MTV treated/MTV control] x 100

Tumor growth and weight changes are measured twice weekly.

For comparisons between vehicle control and TEAD inhibitor treated groups, independent sample t-tests are used for statistical analysis.

MSTO-211H In Vivo Efficacy Study 6-8 week old SCID mice (CRL) are inoculated subcutaneously on the right flank with 5 x ¹⁰ MSTO-211H human mesothelioma tumor cells. Monitor tumor growth twice a week using calipers and calculate mean tumor volume (MTV) using the formula V=W2×L/2.

Once MTV reached approximately 150-200 ^mm3 , animals were randomized into treatment groups (n = 6-8/group) and treated with either vehicle (5% DMSO + 95% PEG 400) or TEAD inhibitor for 22 hours. Administer orally (PO) on a once daily (QD) schedule for ~25 days.

Randomization and treatment begin on day 0, and percent tumor growth inhibition is calculated on the last day of the study (the day the control MTV reaches the maximum allowed tumor volume) and the following calculations are performed.

%TGI = 100 - [MTV treated/MTV control] x 100

Tumor growth and weight changes are measured twice weekly.

For comparisons between vehicle control and TEAD inhibitor treated groups, independent sample t-tests are used for statistical analysis.

Example 7. TEAD Selectivity Assays The TEAD target selectivity profile of the TEAD inhibitor compounds described herein is based on TEAD isoforms or variants, such as human TEAD1 (UniProt KB ID P28347-1 (SEQ ID NO: 1)), human TEAD2 ( To monitor the interaction of UniProtKB ID Q15562 (SEQ ID NO: 2)), human TEAD3 (UniProtKB ID Q99594 (SEQ ID NO: 3)), human TEAD4 (UniProtKB ID Q15561 (SEQ ID NO: 4)), and YAP1 or TAZ can be measured by any of the exemplary assays provided herein designed to. Although co-immunoprecipitation techniques can be used to monitor protein-protein interactions, increasing throughput is difficult based on the basic technology required. Therefore, using alternative but complementary assays, different TEAD isoforms or variants, e.g. ), human TEAD3 (UniProtKB ID Q99594 (SEQ ID NO: 3)), and human TEAD4 (UniProtKB ID Q15561 (SEQ ID NO: 4)), and YAP1 (or TAZ) are monitored.

The first exemplary assay is an in vitro biochemical fluorescence polarization assay using recombinantly expressed and purified YAP binding domains of individual TEAD isoforms and fluorescently labeled peptides derived from the primary sequence of YAP1 ( Bum-Erdene et al., Cell Chem Biol. 2019 Mar 21;26(3):378-389.e13, the entire contents of which are incorporated herein by reference). Fluorescent peptides and potency are measured by incubating compounds with individual TEAD isoform proteins and quantifying peptide displacement.

A second exemplary assay is a cell-based assay using a split luciferase reporter system (Hall et al., ACS Chem. Biol. 2012, 7, 11, 1848-1857, the entire contents of which are incorporated herein by reference. ). Briefly, the YAP-binding domain of each TEAD isoform is transiently co-expressed in HEK293 cells with the TEAD-binding domain or either YAP1 or TAZ, and the proximity of the two chimeric gene fusion products is monitored by luciferase activity (Nouri et al. Cancers (Basel). 2019 Oct 19;11(10), the entire contents of which are incorporated herein by reference). Compounds that interfere with the interaction of TEAD isoforms with YAP1 (or TAZ) reduce the resulting luciferase activity compared to vehicle-treated controls. In a process similar to fluorescence polarization assays, these chimeric gene fusions are recombinantly expressed in bacterial or insect cells and used as in vitro biochemical assays with luciferase readouts similar to cell-based assays.

Another exemplary assay is described by Karatas et al., the entire contents of which are incorporated herein by reference. , “Discovery of Covalent Inhibitors Targeting the Transcriptional Enhanced Associate Domain (TEAD) Central Pocket,” J. Med. Chem. A thiol conjugation assay that monitors the prevention of covalent binding of a fluorescent initiation probe to a cysteine within the central pocket of a TEAD isoform by a small molecule, as described in 2020. Briefly, the thiol-reactive profluorescent probe N-(4-(7-diethylamino-4-methylcoumarin-3-yl)phenyl)maleimide (CPM) is used. Fluorescence at CPM is quenched by maleimide substitution on the phenyl group, which modulates the resonance between the coumarin carbonyl and the 7-amino group. However, upon reaction with thiols, CPM fluorescence increases dramatically. CPM can be used to inhibit TEAD because reaction of free cysteine residues in the TEAD central pocket generates a fluorescent signal, and small molecules that bind with measurable potency to the TEAD central pocket prevent covalent labeling of cysteine by CPM. Can be investigated. Therefore, in the thiol conjugation assay, fluorescence is measured by adding freshly prepared TEAD protein solution and incubating with test compound in assay buffer at room temperature, followed by addition of CPM solution (Ex/Em: 380 /470nm). Any test compound that binds to the TEAD central pocket with measurable potency will exhibit less fluorescence or inhibition of fluorescence compared to a compound that does not bind to the TEAD central pocket.

Example 8. Inhibition of Malignant Mesothelioma Tumor Cell Growth The tumor cell growth inhibition activity of the TEAD inhibitors described herein is investigated in the NCI-H2052 mesothelioma cell line harboring the NF2 mutation. The cell line is selected based, in part, on its mutational status and the ability of siRNA directed against YAP, TAZ, or TEAD1-TEAD4 to inhibit cell proliferation. The nuclear localization of YAP at confluence is also taken into account. 10,000 cells/well are placed in 96-well black plates, including clear flat-bottomed TC-treated imaging plates, in regular medium containing serum and replaced the next day with starvation medium containing 1% serum. After one day of growth in starvation medium, cells are incubated with TEAD inhibitor compound. The starting concentration is 30 μM and serial dilutions in DMSO and medium are made to 0.1 μM to achieve a final DMSO concentration of 0.5%. Cells are then allowed to grow for 3 days, after which EdU (Invitrogen, Molecular Probe)) is added to each well at a final concentration of 10 mM and cells are returned to the incubator for an additional 24 hours. Remove the starvation medium and add 100 μL of PFA 4% containing Hoechst dye to each well to fix the cells. The cells are then permeabilized by incubating the plates for 15 minutes at room temperature, washing twice with PBS, and adding 100 μL per well of Triton-100 containing 0.3% BSA. After 20 minutes, cells are washed with PBS and EdU detection is performed according to the manufacturer's instructions. Image acquisition is performed using, for example, ImageXpress Micro and analyzed using MetaXpress software (Molecular Device).

While a number of embodiments of the invention have been described, it will be obvious that the basic examples may be modified to provide other embodiments utilizing the compounds and methods of the invention. It is therefore to be understood that the scope of the invention is to be defined by the specification and appended claims, rather than by the specific embodiments set forth in the examples.

Claims (53)

A compound of formula I,
or a pharmaceutically acceptable salt thereof.
[In the formula,
L ¹ is a covalent bond or a C _1-6 divalent straight or branched hydrocarbon chain (where 1, 2, or 3 methylene units of the chain are independently -N(R )-, -O-, or -C(O)-),
Ring A may each be substituted,
selected from
Ring B is
selected from
Each ^RW is independently

selected from
Each R ² is independently -OR, -C(O)NR ₂ , optionally substituted -C _1-6 aliphatic,
selected from
each Y is independently N or ^CR5 ;
each R ³ is independently H or an optionally substituted -C _1-6 aliphatic;
Each R ⁴ is independently -S(O) ₂ NR ₂ , -S(O) ₂ R, -C(O)NR ₂ , -C(O)R, or optionally substituted -C _{1 -6} aliphatic,
Each R ⁵ is independently R, -CN, -C(O)R, -C(O)NR ₂ , or 1 to 2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. an optionally substituted 5- to 6-membered heteroaryl having
each m is independently 0, 1, or 2;
p is 0, 1, or 2;
Each R is independently H, optionally substituted -C _1-6 aliphatic, optionally substituted 3- to 8-membered saturated or partially unsaturated monocyclic carbocyclyl, or nitrogen, oxygen or an optionally substituted 3- to 8-membered saturated or partially unsaturated monocyclic heterocyclyl having 1 to 2 heteroatoms independently selected from sulfur. ] Claim: L ¹ is a C _1-6 divalent straight or branched hydrocarbon chain, wherein 1, 2 or 3 methylene units of said chain are substituted with -N(R)- Item 1. The compound according to item 1, or a pharmaceutically acceptable salt thereof. Ring A may be substituted
The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof. Ring A may be substituted

The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof. Ring A may be substituted

The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof. Ring A may be substituted

The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof. Ring B is
The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof. Each R ^w is independently

A compound according to any one of claims 1 to 7, selected from: or a pharmaceutically acceptable salt thereof. A compound according to claim 1, which is a compound of the formula:
or a pharmaceutically acceptable salt thereof.
[wherein each R ¹ is independently R, halogen, -CN, -C(O)R, -C(O)NR ₂ , -OR, -SR, -S(O) ₂ NR ₂ , or -S(O) ₂ R, and each n is independently 0, 1, 2, or 3. ] A compound according to claim 1, which is a compound of the formula:
or a pharmaceutically acceptable salt thereof.
[wherein each R ¹ is independently R, halogen, -CN, -C(O)R, -C(O)NR ₂ , -OR, -SR, -S(O) ₂ NR ₂ , or -S(O) ₂ R, and each n is independently 0, 1, 2, or 3. ] A compound according to claim 1, which is a compound of the formula:
or a pharmaceutically acceptable salt thereof.
[wherein each R ¹ is independently R, halogen, -CN, -C(O)R, -C(O)NR ₂ , -OR, -SR, -S(O) ₂ NR ₂ , or -S(O) ₂ R, and each n is independently 0, 1, 2, or 3. ] A compound according to claim 1, which is a compound of the formula:
or a pharmaceutically acceptable salt thereof.
[wherein each R ¹ is independently R, halogen, -CN, -C(O)R, -C(O)NR ₂ , -OR, -SR, -S(O) ₂ NR ₂ , or -S(O) ₂ R. ] A compound according to claim 1, which is a compound of the formula:

or a pharmaceutically acceptable salt thereof.
[wherein each R ¹ is independently R, halogen, -CN, -C(O)R, -C(O)NR ₂ , -OR, -SR, -S(O) ₂ NR ₂ , or -S(O) ₂ R, and each n is independently 0, 1, 2, or 3. ] A compound according to claim 1, which is a compound of the formula:

or a pharmaceutically acceptable salt thereof.
[wherein each R ¹ is independently R, halogen, -CN, -C(O)R, -C(O)NR ₂ , -OR, -SR, -S(O) ₂ NR ₂ , or -S(O) ₂ R, and each n is independently 0, 1, 2, or 3. ] A compound according to claim 1, which is a compound of the formula:
or a pharmaceutically acceptable salt thereof.
[wherein each R ¹ is independently R, halogen, -CN, -C(O)R, -C(O)NR ₂ , -OR, -SR, -S(O) ₂ NR ₂ , or -S(O) ₂ R, and each n is independently 0, 1, 2, or 3. ] A compound according to any one of the preceding claims selected from the compounds of Table 1, or a pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising a compound according to any one of the preceding claims, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle. 18. A method for treating cancer in a patient, the method comprising: administering a compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 17 to said patient. said method, comprising administering to said person. 19. The method of claim 18, wherein the cancer is associated with increased TEAD expression. 20. The method of claim 18 or 19, wherein the cancer is associated with increased TEAD activity. 18. A method for inhibiting cancer progression in a patient, the method comprising administering to the patient a compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a medicament according to claim 17. The method comprising administering the composition. 22. The method of claim 21, wherein the cancer is associated with increased TEAD expression. 23. The method of claim 21 or 22, wherein the cancer is associated with increased TEAD activity. 17. A method of treating a patient with a disease or disorder associated with increased expression of TEAD, comprising administering to a patient in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 16. or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 17. 17. A method of treating a patient with a disease or disorder associated with increased TEAD activity, comprising administering to a patient in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 16. or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 17. 17. A method of treating a disease or disorder in which inhibition of TEAD activity is beneficial, comprising administering to a patient a therapeutically effective amount of a compound according to any one of claims 1 to 16, or a compound thereof. 18. The method comprising administering a pharmaceutically acceptable salt or a pharmaceutical composition according to claim 17. 17. A method of treating a disease or disorder in which inhibition of the Hippo pathway would be beneficial, comprising administering to a patient in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 16, or 18. The method comprising administering a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 17. 28. A method according to any one of claims 24 to 27, wherein the disease or disorder is a cell proliferative disorder. 29. The method of claim 28, wherein the cell proliferative disorder is cancer. The method according to any one of claims 18 to 23 and 29, wherein the cancer is a cancer in which YAP is localized in the nucleus of the cancer cell. 26. The method of any one of claims 19, 20, and 22-25, wherein the increased TEAD expression or TEAD activity is an increased TEAD1 expression or TEAD1 activity. 26. The method of any one of claims 19, 20, and 22-25, wherein the increased TEAD expression or TEAD activity is an increased TEAD2 expression or TEAD2 activity. 26. The method of any one of claims 19, 20, and 22-25, wherein the increased TEAD expression or TEAD activity is an increased TEAD3 expression or TEAD3 activity. 26. The method of any one of claims 19, 20, and 22-25, wherein the increased TEAD expression or TEAD activity is an increased TEAD4 expression or TEAD4 activity. The increase in TEAD expression or TEAD activity is an increase in TEAD1 expression or TEAD1 activity; an increase in TEAD2 expression or TEAD2 activity; an increase in TEAD3 expression or TEAD3 activity; an increase in TEAD4 expression. , or an increase in TEAD4 activity; or a combination of any of these. Use of a compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 17, for treating cancer in a patient. 37. The use according to claim 36, wherein said cancer is associated with increased expression of TEAD. 38. The use according to claim 36 or 37, wherein the cancer is associated with increased TEAD activity. Use of a compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 17, for inhibiting the progression of cancer in a patient. 40. The use according to claim 39, wherein the cancer is associated with increased TEAD expression. 41. Use according to claim 39 or 40, wherein the cancer is associated with increased TEAD activity. A compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 17, for treating a disease or disorder associated with increased expression of TEAD. use of things. A compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 17, for treating a disease or disorder associated with increased TEAD activity. use of things. A compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a medicament according to claim 17, for treating a disease or disorder in which inhibition of TEAD activity is beneficial. Use of the composition. A compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 17, for treating a disease or disorder in which Hippo pathway inhibition is beneficial. use of things. Use according to any one of claims 42 to 45, wherein the disease or disorder is a cell proliferative disorder. 47. The use according to claim 46, wherein said cell proliferative disorder is cancer. The use according to any one of claims 36 to 41 and 47, wherein the cancer is a cancer in which YAP is localized in the nucleus of the cancer cell. 44. The use according to any one of claims 37, 38, and 40-43, wherein the increased TEAD expression or TEAD activity is an increased TEAD1 expression or TEAD1 activity. 44. The use according to any one of claims 37, 38, and 40-43, wherein the increased TEAD expression or TEAD activity is an increased TEAD2 expression or TEAD2 activity. 44. The use according to any one of claims 37, 38, and 40-43, wherein said increased TEAD expression or increased TEAD activity is increased TEAD3 expression or increased TEAD3 activity. 44. The use according to any one of claims 37, 38, and 40-43, wherein the increased TEAD expression or TEAD activity is an increased TEAD4 expression or TEAD4 activity. The increase in TEAD expression or TEAD activity is an increase in TEAD1 expression or TEAD1 activity; an increase in TEAD2 expression or TEAD2 activity; an increase in TEAD3 expression or TEAD3 activity; an increase in TEAD4 expression. or an increase in TEAD4 activity; or a combination of any of these.