KR20230131189A - TEAD inhibitors and uses thereof - Google Patents

Abstract

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

Description

TEAD inhibitors and uses thereof

sequence list

This application contains a sequence listing that has been submitted electronically in ASCII format, the entire contents of which are incorporated herein by reference. The ASCII copy created on November 30, 2021 is named 187446_SL.txt and is 23,551 bytes in size.

Technical field of invention

The present invention relates to compounds and methods useful for inhibition of Transcriptional Enhancer Associate Domain (TEAD). The invention also provides pharmaceutically acceptable compositions comprising compounds of the invention and methods of using such compositions in the treatment of various diseases, disorders and conditions as described herein.

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

The Hippo signaling cascade is an important pathway for cancer biosynthesis and tumor maintenance. The Hippo pathway is significantly mutated across many cancer indications through loss of function mutations in genes such as NF2. These pro-tumor mutations are constitutive of the downstream transcriptional coactivators YAP and TAZ, which induce the expression of many pro-survival and proliferative genes through essential interactions with members of the TEAD protein family. It leads to activation. Additionally, this derepressed transcriptional program leads to enhanced immune suppression in the tumor microenvironment. As described herein, to target this oncogenic pathway, novel small molecule inhibitors were identified that downregulate YAP- and TAZ-dependent transcription by selectively binding to TEAD and interfering with its interaction with YAP and TAZ. As demonstrated herein, these TEAD inhibitors prevent TEAD palmitoylation, which is important for the interaction between YAP and TEAD. Additionally, the TEAD inhibitors described herein inhibit YAP-dependent in vitro proliferation (i.e., Hippo pathway-deficient cancer cell lines) but not 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. Follow-up experiments in vivo demonstrate that the TEAD inhibitors described herein downregulate YAP-dependent genes in human tumor xenografts following oral administration. Additionally, the TEAD inhibitors described herein demonstrate 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 discovered that the compounds of the present invention, and pharmaceutically acceptable compositions thereof, are effective as TEAD inhibitors. In one aspect, the invention provides a compound of formula ( I) :

In the above formula, each variable is independently as defined herein and as described in the embodiments herein.

The compounds of the present invention, and pharmaceutically acceptable salts and compositions thereof, are useful for treating a variety of diseases, disorders or conditions associated with TEAD. Such diseases, disorders or conditions include cell proliferative disorders (eg, cancer as described herein).

Figure 1 shows a schematic diagram of Hippo pathway signaling.

1. General Description of Certain Embodiments of the Invention:

The compounds of the present invention, and pharmaceutical salts and compositions thereof, are useful as inhibitors of TEAD. Without wishing to be bound by any particular theory, it is believed that the compounds of the present invention, and pharmaceutical compositions thereof, inhibit the activity of TEAD, thereby treating diseases, disorders or conditions associated with TEAD, such as cancer.

In one aspect, the invention provides a compound of formula ( I) :

In the above equation,

L ¹ is a covalent bond, or a C _1-6 divalent straight or branched hydrocarbon chain, wherein 1, 2 or 3 methylene units of the chain are -N(R)-, -O-, or -C(O )-, independently and optionally replaced by;

Ring A is is selected from, each of which is optionally substituted;

Ring B is

is selected from;

Each R ^w is independently is selected from;

Each R ² is independently -OR, -C(O)NR ₂ , optionally substituted -C _1-6 aliphatic, is selected from;

Each Y is independently N or CR ⁵ ;

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 an optionally substituted 5- having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. It is a 6-membered heteroaryl;

Each m is independently 0, 1, or 2;

p is 0, 1, or 2;

Each R is independently selected from H, optionally substituted -C _1-6 aliphatic, optionally substituted 3-8 membered saturated or partially unsaturated monocyclic carbocyclyl, or nitrogen, oxygen or sulfur. It is an optionally substituted 3-8 membered saturated or partially unsaturated monocyclic heterocyclyl having 2 heteroatoms.

2. Compounds and definitions:

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

As used herein, the term “aliphatic” or “aliphatic group” refers to a substituted or unsubstituted hydrocarbon chain, straight (i.e., unbranched) or branched, either fully saturated or containing one or more units of unsaturation, or A monocyclic, non-aromatic (also referred to herein as “carbocycle,” “cycloaliphatic,” or “cycloalkyl”), fully saturated or containing one or more units of unsaturation, but having a single point of attachment to the remainder of the molecule. It means hydrocarbon or bicyclic hydrocarbon. Unless otherwise specified, aliphatic groups contain 1 to 6 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1 to 5 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1 to 4 aliphatic carbon atoms. In another embodiment, the aliphatic group contains 1 to 3 aliphatic carbon atoms, and in another embodiment, the aliphatic group contains 1 to 2 aliphatic carbon atoms. In some embodiments, an “alicyclic” (or “carbocycle” or “cycloalkyl”) is a monocyclic, non-aromatic, that is fully saturated or contains one or more units of unsaturation, but has a single point of attachment to the remainder of the molecule. C ₃ -C ₆ refers to hydrocarbons. Suitable aliphatic groups include linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups, and hybrids thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl. However, it is not limited to these.

As used herein, the term "bicyclic ring" or "bicyclic ring system" refers to any bicyclic ring system, i.e., saturated or having one or more unsaturated units common between two rings of the ring system. refers to a carbocyclic or heterocyclic group having one or more atoms of Accordingly, the term includes any acceptable ring fusion, 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 bicycle. These heteroatoms may be present in ring junctions and may be optionally substituted, and include nitrogen (including N-oxides), oxygen, sulfur (including oxidized forms such as sulfones and sulfonates), and phosphorus (including oxidized forms such as phosphates). ), boron, etc. In some embodiments, a bicyclic group has 7-12 ring members and 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. As used herein, the term “bridged bicyclic” refers to any saturated or partially unsaturated bicyclic ring system having at least one bridge, either carbocyclic or heterocyclic. As defined by IUPAC, a “bridge” is an unbranched chain of atoms, or an atom or valence bond connecting two crosslinkheads, where a “bridgehead” is a ring bonded to three or more backbone atoms (excluding hydrogen). It is an arbitrary skeletal atom of the system. In some embodiments, the bridged bicyclic group has 7-12 ring members and 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Such bridged bicyclic groups are well known in the art and include those groups specified 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 are optionally substituted with one or more substituents as specified for aliphatic groups. Additionally or alternatively, any substitutable nitrogen of the bridged bicyclic group is optionally substituted. Exemplary bicyclic rings include:

.

Exemplary cross-linked bicyclics include:

.

The term “lower alkyl” refers to a C _1-4 straight or branched alkyl group. Exemplary lower alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and tert- butyl.

The term “lower haloalkyl” refers to a C _1-4 straight or branched alkyl group substituted with one or more halogen atoms.

The term "heteroatom" refers to oxygen, sulfur, nitrogen, phosphorus or silicon (any oxidized form of nitrogen, sulfur, phosphorus or silicon; any quaternized form of a basic nitrogen; or a replaceable nitrogen of a heterocyclic ring, e.g. For example, (as in 3,4-dihydro-2H-pyrrolyl) means one or more of N, NH (as in pyrrolidinyl), NR ⁺ (as in N-substituted pyrrolidinyl).

As used herein, the term “unsaturated” 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” refers to a straight or branched divalent alkylene chain as defined herein. , alkenylene, and alkynylene chain.

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 to 6, 1 to 4, 1 to 3, 1 to 2, or 2 to 3. am. A substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced by a substituent. Suitable substituents include those described below for substituted aliphatic groups.

The term “alkenylene” refers to a divalent alkenyl group. Substituted alkenylene chains are polymethylene groups containing at least one double bond in which one or more hydrogen atoms are replaced by a substituent. Suitable substituents include those described below for substituted aliphatic groups.

As used herein, the term “cyclopropylenyl” has the following structure: 2 refers to a cyclopropyl group.

The term “halogen” means F, Cl, Br or I.

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

The terms "heteroaryl" and "heteroar-", used alone or as part of a larger moiety, such as "heteroaralkyl" or "heteroaralkoxy", refer to a group of 5-10 ring atoms, preferably having 5, 6 or 9 ring atoms; Has 6, 10, or 14 π electrons shared in a cyclic configuration; Refers to a group having 1-5 heteroatoms in addition to carbon atoms. The term “heteroatom” refers to nitrogen, oxygen, or sulfur and includes nitrogen or sulfur in any oxidized form, and basic nitrogen in any quaternized form. Heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, and pyrazolyl. Includes diyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. As used herein, the terms "heteroaryl" and "heteroar-" also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, wherein the radical or point of attachment is a heterocyclic ring. It is on an aromatic ring. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthala. Zinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido [2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl groups can be mono- or bicyclic. The term “heteroaryl” may be used interchangeably with the terms “heteroaryl ring,” “heteroaryl group,” or “heteroaromatic,” any of which terms include rings that are optionally substituted. The term “heteroaralkyl” refers to an alkyl group substituted with heteroaryl, where the alkyl and heteroaryl moieties are independently and optionally substituted.

As used herein, the terms “heterocycle,” “heterocyclyl,” “heterocyclic radical,” and “heterocyclic ring” are used interchangeably and are saturated or partially unsaturated and contain additional carbon atoms. refers to a stable 5-7 membered monocyclic or 7-10 membered bicyclic heterocyclic moiety having one or more, preferably 1-4 heteroatoms as defined above. When used in relation to the ring atoms of a heterocycle, the term “nitrogen” includes substituted nitrogen. For example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur, or nitrogen, nitrogen is N (as in 3,4-dihydro-2H-pyrrolyl), (pyrrolyl NH (as in dinyl), or ⁺ NR (as in N-substituted pyrrolidinyl).

Heterocyclic rings can be attached to any heteroatom or carbon atom pendant thereof to create a stable structure, and any ring atom can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, deca Includes hydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms “heterocycle”, “heterocyclyl”, “heterocyclyl ring”, “heterocyclic group”, “heterocyclic moiety” and “heterocyclic radical” are used interchangeably herein; Also included are groups in which the heterocyclyl ring is fused to one or more aryl, heteroaryl or cycloaliphatic rings, such as indolinyl, 3H - indolyl, chromanyl, phenanthridinyl or tetrahydroquinolinyl. Heterocyclyl groups can be mono- or bicyclic. The term “heterocyclylalkyl” refers to an alkyl group substituted with heterocyclyl, wherein the alkyl and heterocyclyl moieties are independently and optionally substituted.

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

As described herein, the compounds of the invention may contain “optionally substituted” moieties. In general, the term "substituted", whether preceded by the term "optionally" or not, means that one or more hydrogens of the specified moiety are replaced by a suitable substituent. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position on the group, and more than one position in any given structure may be substituted with more than one substituent selected from the designated group. Where possible, the substituents may be the same or different at all positions. Combinations of substituents contemplated by the present invention are preferably those that form stable or chemically feasible compounds. As used herein, the term “stable” means that a compound does not change substantially when subjected to conditions that permit its production, detection, and, in certain embodiments, recovery, purification, and use for one or more of the purposes disclosed herein. Refers to compounds that do not work.

Each optional substituent on a replaceable carbon is halogen; -(CH ₂ ) _0-4 R°; -(CH ₂ ) _0-4 OR°; -O(CH ₂ ) _0-4 R ^o , -O-(CH ₂ ) _0-4 C(O)OR°; -(CH ₂ ) _0-4 CH(OR°) ₂ ; -(CH ₂ ) _0-4 SR°; -(CH ₂ ) _0-4 Ph which may be substituted by R°; -(CH ₂ ) _0-4 O(CH ₂ ) _0-1 Ph which may be substituted by R°; -CH=CHPh, which can be replaced by R°; -(CH ₂ ) _0-4 O(CH ₂ ) _0-1 -pyridyl which may be substituted by R°; -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) _2R °; -P(O)R° ₂ ; -OP(O)R° ₂ ; -OP(O)(OR°) ₂ ; SiR° ₃ ; -(C _1-4 straight or branched alkylene)ON(R°) ₂ ; or -(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 -(5-6 membered heteroaryl ring), or 5-6 membered heteroaryl ring. an aryl ring that is saturated, partially unsaturated, or has 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or notwithstanding the above definition, two independently occurring R°, together with their intervening atom(s), may be a 3-12 membered group having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Forms a saturated, partially unsaturated, or aryl mono- or bicyclic ring, which may be substituted by a divalent substituent on the saturated carbon atom of R° selected from =O and =S; Each R° is halogen, -(CH ₂ ) _0-2 R ^ㆍ , -(haloR ^ㆍ ), -(CH ₂ ) _0-2 OH, -(CH ₂ ) _0-2 OR ^ㆍ , -(CH ₂ ) _0-2 CH(OR ^ㆍ ) ₂ ; -O(haloR ^ㆍ ), -CN, -N ₃ , -(CH ₂ ) _0-2 C(O)R ^ㆍ , -(CH ₂ ) _0-2 C(O)OH, -(CH ₂ ) _{0 -2} C(O)OR ^ㆍ , -(CH ₂ ) _0-2 SR ^ㆍ , -(CH ₂ ) _0-2 SH, -(CH ₂ ) _0-2 NH ₂ , -(CH ₂ ) _0-2 NHR ^ㆍ , -(CH ₂ ) _0-2 NR ^ㆍ ₂ , -NO ₂ , -SiR ^ㆍ ₃ , -OSiR ^ㆍ ₃ , -C(O)SR ^ㆍ _, -(C _1-4 straight or branched alkylene)C (O)OR ^ㆍ , or -SSR ^ㆍ is optionally substituted with a monovalent substituent independently selected from.

Each R ^· is C _1-4 aliphatic, -CH ₂ Ph, -O(CH ₂ ) _0-1 Ph, or a 5-6 membered saturated group having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. , partially unsaturated, or an aryl ring, wherein each R ^· is unsubstituted or, if preceded by halo, substituted only by one or more halogens; or wherein optional substituents on the saturated carbon are =O, =S, =NNR ^* ₂ , =NNHC(O)R ^* , =NNHC(O)OR ^* , =NNHS(O) ₂ R ^* , =NR ^* , = a divalent substituent independently selected from NOR ^* , -O(C(R ^* ₂ )) _2-3 O-, or -S(C(R ^* ₂ )) _2-3 S-, or "optionally substituted" The divalent substituent bonded to the adjacent substitutable carbon of the group is -O(CR ^* ₂ ) _2-3 O-, wherein each independently occurring R ^* is hydrogen, C _1-6 aliphatic, or unsubstituted 5-6 membered saturated, partially unsaturated, or aryl having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. is selected from a ring.

If R ^* is C _1-6 aliphatic, R ^* is halogen, -R ^ㆍ , -(haloR ^ㆍ ), -OH, -OR ^ㆍ , -O(haloR ^ㆍ ), -CN, -C(O)OH , -C(O)OR ^ㆍ , -NH ₂ , -NHR ^ㆍ , -NR ^ㆍ ₂ , or -NO ₂ , where each R ^ㆍ is C _1-4 aliphatic, -CH ₂ Ph, -O (CH ₂ ) is independently selected from _0-1 Ph, or a 5-6 membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, wherein each R ^· is If not substituted, or if preceded by halo, it is substituted only with one or more halogens.

Optional substituents on the replaceable nitrogen are 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 ^† and; wherein each R ^† is independently hydrogen, C _1-6 aliphatic, unsubstituted -OPh, or 5-6 membered unsubstituted saturated with 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, Partially unsaturated, or an aryl ring, or two independently occurring R ^† , together with their intervening atom(s), are an unsubstituted 3- group having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Forms a 12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring; Here, if R ^† is C _1-6 aliphatic, R ^† is halogen, -R ^ㆍ , -(haloR ^ㆍ ), -OH, -OR ^ㆍ , -O(haloR ^ㆍ ), -CN, -C(O) optionally substituted with OH, -C(O)OR ^ㆍ , -NH ₂ , -NHR ^ㆍ , -NR ^ㆍ ₂ , or -NO ₂ , where each R ^ㆍ is C _1-4 aliphatic, -CH ₂ Ph, - O(CH ₂ ) _0-1 Ph, or a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, where each R ^· is unsubstituted Or, if halo precedes it, it is replaced only by one or more halogens.

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

Salts derived from suitable bases include alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Additional pharmaceutically acceptable salts may be prepared using non-toxic ammoniums, quaternary ammoniums, and counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates, as appropriate. Contains the amine cation formed.

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

As used herein, the term “inhibitor” or “TEAD inhibitor” or “TEAD antagonist” is defined as a compound that binds and/or inhibits TEAD with measurable affinity. 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 about 15%, or at least greater than the signal measured using a negative control under comparable conditions. About 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least About 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or At least about 100% lower. The potency of an inhibitor is usually defined by its IC ₅₀ value (the concentration required to inhibit 50% of the agonist response, or half the maximum inhibitory concentration). The lower the IC ₅₀ value, the greater the potency of the antagonist, and the lower the concentration required to inhibit the 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. have

As used herein, the terms "measurable affinity" and "measurably inhibit" refer to a sample comprising a compound of the invention, or a composition thereof, and a TEAD, and to a TEAD in the absence of the compound, or composition thereof. means a measurable change or inhibition in TEAD activity between equivalent samples comprising.

3. Description of Exemplary Embodiments:

In one aspect, the invention provides a compound of formula ( I) :

In the above equation,

L ¹ is a covalent bond, or a C _1-6 divalent straight or branched hydrocarbon chain, wherein 1, 2 or 3 methylene units of the chain are -N(R)-, -O-, or -C(O )-, independently and optionally replaced by;

Ring A is is selected from, each of which is optionally substituted;

Ring B is is selected from;

Each R ^w is independently is selected from;

Each R ² is independently -OR, -C(O)NR ₂ , optionally substituted -C _1-6 aliphatic, is selected from;

Each Y is independently N or CR ⁵ ;

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 an optionally substituted 5- having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. It is a 6-membered heteroaryl;

Each m is independently 0, 1, or 2;

p is 0, 1, or 2;

Each R is independently selected from H, optionally substituted -C _1-6 aliphatic, optionally substituted 3-8 membered saturated or partially unsaturated monocyclic carbocyclyl, or nitrogen, oxygen or sulfur. It is an optionally substituted 3-8 membered saturated or partially unsaturated monocyclic heterocyclyl having 2 heteroatoms.

As generally defined above, L ¹ is a covalent bond, or a C _1-6 divalent straight or branched hydrocarbon chain, wherein 1, 2 or 3 methylene units of the chain are -N(R)-, - is independently and optionally replaced by 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, wherein 1, 2, or 3 methylene units of the chain are independently and optionally replaced with -N(R)-.

In some embodiments, L ¹ is a C _1-6 divalent straight or branched hydrocarbon chain, wherein 1, 2, or 3 methylene units of the chain are independently and optionally replaced with -O-.

In some embodiments, L ¹ is a C _1-6 divalent straight or branched hydrocarbon chain, wherein 1, 2, or 3 methylene units of the chain are independently and optionally replaced with -C(O)-.

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 am. In some embodiments, L ¹ is am. In some embodiments, L ¹ is am. In some embodiments, L ¹ is am. In some embodiments, L ¹ is -CH=CH-. In some embodiments, L ¹ is am. In some embodiments, L ¹ is am. In some embodiments, L ¹ is -NH-C(O)-.

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

As generally defined above, Ring A is is selected from, each of which is optionally substituted.

In some embodiments, Ring A is optionally substituted am.

In some embodiments, Ring A is optionally substituted am.

In some embodiments, Ring A is optionally substituted am.

In some embodiments, Ring A is optionally substituted am.

In some embodiments, Ring A is optionally substituted am.

In some embodiments, Ring A is optionally substituted am.

In some embodiments, Ring A is optionally substituted am.

In some embodiments, Ring A is optionally substituted am.

In some embodiments, Ring A is optionally substituted am.

In some embodiments, Ring A is optionally substituted am.

In some embodiments, Ring A is optionally substituted am.

In some embodiments, Ring A is optionally substituted am.

In some embodiments, Ring A is

is selected from, where 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, wherein each R is independently as defined herein and as described in the embodiments herein.

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 1-6 halogens, 3-8 membered saturated or partial, optionally substituted with 1-6 halogens. unsaturated monocyclic carbocyclyl, or 3-8 membered saturated or partially unsaturated monocyclic heteroatoms having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, optionally substituted with 1-6 halogens. It's a cycle reel.

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, or am.

In some embodiments, each R ¹ is independently selected from those shown 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 , alone and in combination, is as defined above and described in the embodiments herein.

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

In some embodiments, Ring A is

am.

In some embodiments, Ring A is is selected from

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

As generally defined above, ring B is wherein each of R ² , R ³ , R ^w , p and R ⁴ , alone and in combination, is as defined herein and described in the embodiments herein.

In some embodiments, Ring B is and wherein each of R ² and R ^w , alone and in combination, is as defined herein and described in the embodiments herein.

In some embodiments, Ring B is and wherein each of R ⁴ and R ^w , alone and in combination, is as defined herein and described in the embodiments herein.

In some embodiments, Ring B is and wherein each of R ² and R ^w , alone and in combination, is as defined herein and described in the embodiments herein.

In some embodiments, Ring B is and wherein each of R ² and R ^w , alone and in combination, is as defined herein and described in the embodiments herein.

In some embodiments, Ring B is and wherein each of R ⁴ and R ^{w ,} alone and in combination, is as defined herein and described in the embodiments herein.

In some embodiments, Ring B is and wherein each of R ² and R ^w , alone and in combination, is as defined herein and described in the embodiments herein.

In some embodiments, Ring B is and wherein each of R ³ and p, alone and in combination, is as defined herein and described in the embodiments herein. In some embodiments, Ring B is am.

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 shown in Table 1 below.

As generally defined above, R ^w is is selected from

In some embodiments, R ^w is am. In some embodiments, R ^w is am. In some embodiments, R ^w is am. In some embodiments, R ^w is am. In some embodiments, R ^w is am. In some embodiments, R ^w is am. In some embodiments, R ^w is am.

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

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

In some embodiments, R ² is -OR. In some embodiments, R ² is -C(O)NR ₂ . In some embodiments, R ² is optionally substituted -C _1-6 aliphatic. In some embodiments, R ² is am. In some embodiments, R ² is am. In some embodiments, R ² is am. In some embodiments, R ² is am. In some embodiments, R ² is am. In some embodiments, R ² is am. In some embodiments, R ² is am.

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

In some embodiments, R ² is -CH ₃ , -CH ₂ CH ₃ ,

is selected from

In some embodiments, R ² is is selected from

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

As generally defined above, each Y is independently N or CR ⁵ .

In some embodiments, Y is N. In some embodiments, Y is CR ⁵ . In some embodiments, Y is CH.

In some embodiments, both Ys are N. In some embodiments, both Y is CR ⁵ . In some embodiments, one Y is N and the other Y is CR ⁵ . In some embodiments, both Y are CH. In some embodiments, one Y is N and the other Y is CH.

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

As generally defined above, each R ³ is independently H, -C(O)R, or optionally substituted -C _1-6 aliphatic, wherein R is defined herein and described in the embodiments herein. It's the same as what happened.

In some embodiments, R ³ is H.

In some embodiments, R ³ is -C(O)R.

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

In some embodiments, R ³ is H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH ₃ , -C(O)CH ₃ , and is selected from

In some embodiments, R ³ is selected from those shown 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 is -C _1-6 aliphatic, wherein 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 optionally substituted -C _1-6 aliphatic.

In some embodiments, R ⁴ is is selected from

In some embodiments, R ⁴ is is selected from

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

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

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, and sulfur.

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

is selected from

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

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

As generally defined above, each m is independently 0, 1, or 2.

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 shown in Table 1 below.

As generally defined above, p is 0, 1, or 2.

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 shown in Table 1 below.

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

In some embodiments, R is H.

In some embodiments, R is -C _1-6 aliphatic, optionally substituted. 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 -CH ₃ . In some embodiments, R is -CH ₂ CH ₃ . In some embodiments, R is -CF ₃ . In some embodiments, R is -CHF ₂ .

In some embodiments, R is an optionally substituted 3-8 membered saturated or partially unsaturated monocyclic carbocyclyl. In some embodiments, R is unsubstituted 3, 4, 5, 6, 7 or 8 membered saturated or partially unsaturated monocyclic carbocyclyl. In some embodiments, R is 3, 4, 5, 6, 7 or 8 substituted 1, 2, 3, 4, 5 or 6 times with -halogen, -CN, -NO ₂ , or -C _1-6 aliphatic. a circularly saturated or partially unsaturated monocyclic carbocyclyl, wherein the -C _1-6 aliphatic is optionally substituted 1, 2, 3, 4, 5 or 6 times with -halogen, -CN, or -NO ₂ . In some embodiments, R is 3, 4, 5, 6, 7 or 8 membered saturated or partially unsaturated monocyclic carbocyclyl substituted 1, 2, 3, 4, 5 or 6 times with halogen. In some embodiments, R is a 3, 4, 5, 6, 7 or 8 membered saturated or partially unsaturated monocyclic carbocycle that is 1, 2, 3, 4, 5 or 6 times substituted with -C _1-6 aliphatic. Ryl, wherein the C _1-6 aliphatic is optionally substituted 1, 2, 3, 4, 5 or 6 times with -halogen. In some embodiments, R is 3, 4, 5, 6, 7 or 8 membered saturated or partially unsaturated monocyclic carbocyclyl substituted 1, 2, 3, 4, 5 or 6 times with -F. In some embodiments, R is a 3, 4, 5, 6, 7 or 8 membered saturated or partially unsaturated monocyclic carbocycle that is 1, 2, 3, 4, 5 or 6 times substituted with -C _1-6 aliphatic. reel, wherein the -C _1-6 aliphatic is optionally substituted 1, 2, 3, 4, 5 or 6 times with -F.

In some embodiments, R is an optionally substituted 3-8 membered saturated or partially unsaturated monocyclic heterocyclyl having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, R is an unsubstituted 3, 4, 5, 6, 7 or 8 membered saturated or partially unsaturated monocyclic heterocyclyl having 1-2 heteroatoms independently selected from nitrogen, oxygen or sulfur. am. In some embodiments, R is a 3, 4, 5, 6, 7 or 8 membered saturated or partially unsaturated monocyclic heterocyclyl having 1-2 heteroatoms independently selected from nitrogen, oxygen or sulfur, which -halogen, -CN, -NO ₂ , or -C _1-6 aliphatic substituted 1, 2, 3, 4, 5 or 6 times, wherein the -C _1-6 aliphatic is -halogen, -CN or -NO ₂ is optionally substituted 1, 2, 3, 4, 5 or 6 times. In some embodiments, R is -3, 4, 5, 6 having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, substituted 1, 2, 3, 4, 5, or 6 times with halogen. , is a 7- or 8-membered saturated or partially unsaturated monocyclic heterocyclyl. In some embodiments, R is 3, 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, substituted 1, 2, 3, 4, 5, or 6 times with -C _1-6 aliphatic. , a 5, 6, 7 or 8 membered saturated or partially unsaturated monocyclic heterocyclyl, wherein the C _1-6 aliphatic is optionally substituted 1, 2, 3, 4, 5 or 6 times with -halogen. In some embodiments, R is 3, 4, 5, 6 having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and substituted 1, 2, 3, 4, 5, or 6 times with -F. , is a 7- or 8-membered saturated or partially unsaturated monocyclic heterocyclyl. In some embodiments, R is 3, 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, substituted 1, 2, 3, 4, 5, or 6 times with -C _1-6 aliphatic. , 5, 6, 7 or 8 membered saturated or partially unsaturated monocyclic heterocyclyl, wherein the -C _1-6 aliphatic is optionally substituted 1, 2, 3, 4, 5 or 6 times with -F. .

In some embodiments, R is

is selected from

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

In some embodiments, the invention provides a compound of the formula:

wherein each of R ¹ , L ¹ , R ^w , Y, m, n, and R ⁵ , alone and in combination, is as defined above and as described in the embodiments herein.

In some embodiments, the invention provides a compound of the formula:

wherein each of R ¹ , L ¹ , R ^w , Y, n, and R ⁵ , alone and in combination, is as defined above and as described in the embodiments herein.

In some embodiments, the invention provides a compound of the formula:

wherein each of R ¹ , L ¹ , R ^w , n, and R ⁵ , alone and in combination, is as defined above and as described in the embodiments herein.

In some embodiments, the invention provides a compound of the formula:

wherein each of R ¹ , R ^w , L ¹ , and R ⁵ , alone and in combination, is as defined above and as described in the embodiments herein.

In some embodiments, the invention provides a compound of the formula:

wherein each of R, R ¹ , L ¹ , R ^w , and n, alone and in combination, is as defined above and as described in the embodiments herein.

In some embodiments, the invention provides a compound of the formula:

wherein each of R, R ¹ , L ¹ , R ^w , and n, alone and in combination, is as defined above and as described in the embodiments herein.

In some embodiments, the invention provides a compound of the formula:

wherein each of R ¹ , L ¹ , R ^w , and n, alone and in combination, is as defined above and as described in the embodiments herein.

In some embodiments, the invention provides a compound of the formula:

wherein each of R ¹ , L ¹ , R ^w , Y, m, n and R ⁵ , alone and in combination, is as defined above and as described in the embodiments herein.

In some embodiments, the invention provides a compound of the formula:

wherein each of R ¹ , L ¹ , R ^w , Y, n, and R ⁵ , alone and in combination, is as defined above and as described in the embodiments herein.

In some embodiments, the invention provides a compound of the formula:

wherein each of R ¹ , L ¹ , R ^w , n, and R ⁵ , alone and in combination, is as defined above and as described in the embodiments herein.

In some embodiments, the invention provides a compound of the formula:

wherein each of R ¹ , L ¹ , R ^w , and R ⁵ , alone and in combination, is as defined above and as described in the embodiments herein.

Without wishing to be bound by any particular theory, the compounds provided herein may modify amino acid residues (e.g., cysteine, lysine, histidine, or covalently modified) present in the binding pocket of a target protein, e.g., TEAD. By covalently binding to other residues, it can irreversibly inhibit the protein. In some embodiments, the warhead group can covalently bind to cysteine. In some embodiments, the warhead group can covalently bind serine. In some embodiments, the warhead group can covalently bind lysine. In some embodiments, the warhead group may covalently bind to Cys359 of hTEAD1, Cys405 of hTEAD1, Cys380 of hTEAD2, Cys368 of hTEAD3, and/or Cys367 of hTEAD4. In some embodiments, the warhead group may covalently bind to Ser356 of hTEAD1, Ser345 and/or Ser377 of hTEAD2, Ser365 of hTEAD3, and/or Ser364 of hTEAD4. In some embodiments, the warhead group may covalently bind to Lys336 of hTEAD1, Lys357 of hTEAD2, Lys345 of hTEAD3, and/or Lys344 of hTEAD4. Representative reference amino acid sequences of human TEAD1, human TEAD2, human TEAD3, and human TEAD4 are UniProt KB ID P28347-1 (SEQ ID NO: 1), UniProtKB ID Q15562 (SEQ ID NO: 2), and UniProtKB ID Q99594 (SEQ ID NO: 3), respectively. ), and UniProtKB ID Q15561 (SEQ ID NO: 4). “Targeting Transcriptional Enhanced Associate Domains (TEADs),” J. Med, the entire contents of which are incorporated herein by reference. Chem. A sequence alignment of the TEAD coactivator binding domains shown in Table 1 of 2018 , 61, 5057-5072 is given below.

Exemplary compounds of the invention are specified in Table 1 below.

Exemplary Compounds

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

In some embodiments, the compounds of the invention are not compounds selected from:

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

4. Uses, formulation and administration:

Pharmaceutically acceptable composition

According to another embodiment, the present invention provides a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant or vehicle. The amount of compound in the compositions of the invention is an amount effective to measurably inhibit TEAD, or variants or mutants thereof, in a biological sample or in a patient. In certain embodiments, the amount of compound in the compositions of the invention is an amount effective to measurably inhibit TEAD, or variants or mutants thereof, in a biological sample or in a patient. In certain embodiments, the compositions of the invention are formulated for administration to a patient in need of such compositions. In some embodiments, the compositions of the invention are formulated for oral administration to a patient.

As used herein, the term “patient” or “subject” 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 destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or vehicles that can be used in the compositions of the invention include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphate, glycine, sorbic acid, potassium sorbate. , mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulosic substances. , polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and wool fat.

“Pharmaceutically acceptable derivative” means any non-toxic salt of a compound of the invention, which, when administered to a recipient, may directly or indirectly provide the compound of the invention or an inhibitory active metabolite or residue thereof; means ester, salt of ester or other derivative.

As used herein, the term “inhibitory active metabolite or residue thereof” means that the metabolite or residue thereof is also an inhibitor of TEAD, or a variant or mutant thereof.

Compositions of the invention can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, bucally, vaginally or via an implanted reservoir. As used herein, the term “parenteral” includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. Preferably, the composition is administered orally, intraperitoneally or intravenously. Sterile injectable forms of the compositions of the present invention may be aqueous or oleaginous suspensions. 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 non-toxic, parenterally acceptable diluent or solvent, for example, a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be used are water, Ringer's solution and isotonic sodium chloride solution. Additionally, sterile, fixed oils are commonly used as solvents or suspending media.

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

Pharmaceutically acceptable compositions of the invention may be administered orally in any orally acceptable dosage form, including but not limited to capsules, tablets, aqueous suspensions, or solutions. For oral tablets, commonly used carriers include lactose and corn starch. Lubricants such as magnesium stearate are also typically added. For oral administration in capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredients are combined with emulsifying agents and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.

Alternatively, the pharmaceutically acceptable compositions of the present invention may be administered in the form of suppositories for rectal administration. Since they are solid at room temperature but liquid at rectal temperature, they can be prepared by mixing the formulation with an appropriate non-irritating excipient that dissolves in the rectum and releases the drug. These substances include cocoa butter, beeswax, and polyethylene glycol.

Pharmaceutically acceptable compositions of the invention may also be administered topically, especially when the target of treatment involves areas or organs readily accessible by topical application, including diseases of the eyes, skin, or lower intestinal tract. It can be. Suitable topical formulations are easily prepared for each of these areas or organs.

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

For topical application, the provided pharmaceutically acceptable compositions may be formulated into a suitable ointment containing the active ingredient suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of the present invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsifying wax, and water. Alternatively, provided pharmaceutically acceptable compositions may be formulated as suitable lotions or creams containing the active ingredients 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 are as a micronized suspension in isotonic, pH adjusted sterile saline, with or without a preservative such as benzylalkonium chloride, or preferably at an isotonic pH. It can be formulated as a solution in conditioned sterile saline. Alternatively, for ophthalmic use, the pharmaceutically acceptable composition can be formulated as an ointment, such as petrolatum.

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

Most preferably, the pharmaceutically acceptable compositions of the invention are formulated for oral administration. These formulations may be administered with or without food. In some embodiments, pharmaceutically acceptable compositions of the invention are administered without food. In another embodiment, the pharmaceutically acceptable compositions of the invention are administered with food.

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

Additionally, the specific dosage and treatment regimen for any particular patient will depend on the activity of the particular compound used, age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the treating physician's It should be understood that the determination and treatment will depend on a variety of factors, including the severity of the particular condition being treated. The amount of a compound of the invention in a composition also depends 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 key regulator of cell proliferation, death, and differentiation. In some embodiments, a key function of the Hippo signaling pathway is the transcriptional coactivator Yes-associated protein (YAP) and its paralogue, transcriptional coactivator with PDZ-binding motif (TAZ; also known as WWTR1). ) is adjusted negatively. The Hippo kinase cascade promotes the cytoplasmic retention and degradation of YAP/TAZ, thereby phosphorylating and inhibiting YAP/TAZ, thereby inhibiting the growth-promoting function regulated under YAP/TAZ control. In the unphosphorylated/dephosphorylated state, YAP, also known as YAP1 or YAP65, is transported with TAZ to the nucleus, where it interacts with transcription factors of the TEAD family, upregulating genes that promote proliferation and migration and inhibit apoptosis. . In some cases, uncontrolled upregulation of these genes involved in proliferation, migration, and anti-apoptosis leads to cancer development. In some cases, overexpression of YAP/TAZ is associated with cancer.

Additional key members of the Hippo signaling pathway are the serine/threonine kinases MST1/2 (homologs of Hippo/Hpo in Drosophila), Lats1/2 (homologs of Warts/Wts) and their adapter protein Sav1 (homologs of Salvador/Sav), respectively. and Mob (MOBKL1A and MOBKL1B; homologues of Mats). Typically, MST1/2 kinases form a complex with the scaffold protein Sav1, which in turn phosphorylates and activates Lats1/2 kinases. Lats1/2 is also activated by the scaffold protein Mob. Afterwards, activated Lats1/2 phosphorylates and inactivates YAP or its paralog TAZ. Phosphorylation of YAP/TAZ leads to nuclear export, cytoplasmic retention, 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 is phosphorylated at amino acid position S127. Phosphorylation of YAP S127 promotes 14-3-3 protein binding and results in cytoplasmic sequestration of YAP. Mutation of YAP at position S127 disrupts its interaction with 14-3-3 and subsequently promotes nuclear translocation.

Additional phosphorylation occurs at amino acid position S381 in YAP. Phosphorylation of YAP at position S381 and on the corresponding site of TAZ primes both proteins for further phosphorylation events by CK1δ/ε at the degradation motif, which then signals for interaction with the β-TRCP E3 ubiquitin ligase. leads to 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 is phosphorylated at amino acid position S89. Phosphorylation of TAZ S89 promotes 14-3-3 protein binding and results in cytoplasmic sequestration of TAZ. Mutation of TAZ at position S89 disrupts its interaction with 14-3-3 and subsequently promotes nuclear translocation.

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) includes F-box family member proteins (e.g., Cdc4), Skp1, cross-linking proteins, and RBX1 and interacts with E2-ubiquitin conjugation enzymes. It is a multiprotein E3 ubiquitin ligase complex containing a small RING Finger domain. In some cases, the F-box family includes more than 40 members, where exemplary members include F-box/WD repeat-containing protein 1A (FBXW1A, βTrCP1, Fbxwl, hsSlimb, plkappaBalpha-E3 receptor subunit) and S -Includes phase kinase-related protein 2 (SKP2). In some embodiments, the SCF complex (e.g., SCF ^βTrCP1 ) interacts with the E1 ubiquitin-activating enzyme and the E2 ubiquitin-conjugating enzyme to catalyze the transfer of ubiquitin to the YAP/TAZ substrate. Exemplary E1 ubiquitin-activating enzymes include those encoded by the following genes: UBA1, UBA2, UBA3, UBA5, UBA5, UBA7, ATG7, NAE1 and SAE1. Exemplary E2 ubiquitin-conjugating enzymes include those encoded by the following genes: UBE2A, UBE2B, UBE2C, UBE2D1, UBE2D2, UBE2D3, UBE2E1, UBE2E2, UBE2E3, UBE2F, UBE2G1, UBE2G2, UBE2H, UBE2I, UBE2J1, UBE2J2, UBE2K, UBE2L3, UBE2L6, UBE2M, UBE2N, UBE20, UBE2Q1, UBE2Q2, UBE2R1, UBE2R2, UBE2S, UBE2T, UBE2U, UBE2V1, UBE2V2, UBE2Z, ATG2, BIRC5 and UFC1. In some embodiments, ubiquitinated YAP/TAZ undergoes further degradation via the 26S proteasome.

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

TEAD and YAP/TAZ interaction

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

Proteomic and biochemical studies have shown that TEAD (TEA domain) transcription factors are palmitoylated at evolutionarily conserved cysteine residues. To test whether mutations affect TEAD1 palmitoylation, three cysteine residues (C53S, C327S, and C359S) were found to be evolutionarily conserved in human TEAD1 and mutated to serines. The C359S mutant showed the greatest loss of palmitoylation, while C327S and C53S also showed reduced palmitoylation. These results suggest that C359 plays an important role in TEAD1 palmitoylation. Additionally, a combination mutation of all three cysteine residues, C53/327/359S (3CS), completely abolished TEAD1 palmitoylation, indicating that these residues are involved in TEAD1 palmitoylation. It was shown that TEAD undergoes PAT-independent autopalmitoylation under physiological concentrations of palmitoy l-CoA. Autopalmitoylation also plays an important role in regulating TEAD-YAP binding 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 regulating the Hippo pathway transcription complex.

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

In some embodiments, the compounds described herein irreversibly inhibit TEAD transcription factors. 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, the compounds described herein covalently bind to a TEAD transcription factor (e.g., TEAD1, TEAD2, TEAD3, or TEAD4). In some embodiments, the compounds described herein irreversibly inhibit the activity of a TEAD transcription factor (e.g., TEAD1, TEAD2, TEAD3, or TEAD4). In some embodiments, the compounds described herein covalently inhibit the activity of a TEAD transcription factor (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4).

In some embodiments, compounds disclosed herein bind TEAD1 and interfere with or inhibit the interaction between YAP and TEAD1. In some embodiments, compounds disclosed herein bind TEAD2 and interfere with or inhibit the interaction between YAP and TEAD2. In some embodiments, compounds disclosed herein bind TEAD3 and interfere with or inhibit the interaction between YAP and TEAD3. In some embodiments, compounds disclosed herein bind TEAD4 and interfere with or inhibit the interaction between YAP and TEAD4.

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

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

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

In some embodiments, the compounds disclosed herein bind TEAD2 at C380 and interfere with or inhibit the interaction between YAP and TEAD2.

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

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

In some embodiments, the compounds disclosed herein bind TEAD3 at C371 and interfere with or inhibit the interaction between YAP and TEAD3. In some embodiments, the compounds disclosed herein bind TEAD3 at C368 and interfere with or inhibit the interaction between YAP and TEAD3. In some embodiments, the compounds disclosed herein bind TEAD3 at C371 and C368 and interfere with or inhibit the interaction between YAP and TEAD3.

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

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

In some embodiments, compounds disclosed herein bind TEAD4 at C367 and interfere with or inhibit the interaction between YAP and TEAD4.

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

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

YAP/TAZ regulation mediated by G-protein/GPCR

In some embodiments, the Hippo pathway is regulated by proteins of the G protein coupled receptor (GPCR) and G protein (also known as guanine nucleotide binding protein) families. G proteins are molecular switches that transmit extracellular stimuli to cells through 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 consists of alpha (G _α ), beta (G _β ), and gamma (G _γ ) subunits. In some cases, there are several classes of G _α subunits: G _q/11 α, G _12/13 α, G _i/o α (G inhibitory, G other), and G _s α (G stimulatory).

In some cases, Giα (G repressive), 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 excitatory) bound GPCRs inhibit YAP/TAZ activity, leading to YAP/TAZ degradation.

In some cases, G _i α (G repressive), G _o α (G other), G _q/11 α, and G _12/13 α bound GPCRs activate YAP/TAZ through inhibition of Lats1/2 activity. do. In contrast, G _s α induces Lats1/2 activity in some embodiments, promoting YAP/TAZ degradation.

G _q family

G _q α (also known as G _q/11 protein) participates in the inositol triphosphate (IP ₃ ) signal transduction pathway and calcium (Ca ²⁺ ) release from intracellular stores through activation of phospholipase C (PLC). do. Activated PLC hydrolyzes phosphatidylinositol 4,5-bisphosphate (PIP ₂ ) to diacylglycerol (DAG) and IP ₃ . In some cases, IP ₃ then diffuses through the cytoplasm into the ER, or in the case of muscle cells, the sarcoplasmic reticulum (SR), where it binds to the inositol trisphosphate receptor (InsP3R), a Ca ²⁺ channel. In some cases, this binding causes the opening of Ca ²⁺ channels, increasing the release of Ca ²⁺ into the cytoplasm.

In some embodiments, the GPCR that interacts with G _q α is 5-hydroxytryptamine receptor (5-HT receptor) types 5-HT ₂ and 5-HT ₃ ; alpha-1 adrenergic receptor; Vasopressin type 1 receptors 1A and 1B; Angiotensin II receptor type 1; calcitonin receptor; histamine H1 receptor; Metabotropic glutamate receptors, group I; muscarinic receptors M ₁ , M ₃ , and M ₅ ; and trace amounts of 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 . G _q protein is encoded by GNAQ. G _q/11 is encoded by GNA11. G _q/14 is encoded by GNA14. G _q/15 is encoded by GNA15.

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

In some cases, mutations or variants of the G _q α gene have been associated with congenital diseases. In some cases, mutations in G _q α have been observed in congenital diseases such as Port-Wine Stain and/or Sturge-Weber Syndrome. In some cases, about 92% of flame nevi cases have mutations in GNAQ. In some cases, about 88% of Sturge-Weber syndrome cases have mutations in GNAQ.

G _12/13 family

G _12/13 α coordinates actin cytoskeleton remodeling in cells and regulates cellular processes through guanine nucleotide exchange factors (GEFs). GEFs participate in the activation of small GTPases, which act as molecular switches in various intracellular signaling pathways. Examples of small GTPases include the Ras-related GTPase superfamily (e.g., the Rho family, such as Cdc42), which are involved in cell differentiation, proliferation, cytoskeletal organization, vesicle trafficking, and nuclear transport.

In some embodiments, GPCRs that interact with G _12/13 α include purinergic receptors (eg, P2Y ₁ , P2Y ₂ , P2Y ₄ , P2Y ₆ ); Muscarinic acetylcholine receptors M1 and M3; Receptors for thrombin [protease-activated receptor (PAR)-1, PAR-2]; thromboxane (TXA2); sphingosine 1-phosphate (eg, S1P ₂ , S1P ₃ , S1P ₄ and S1P ₅ ); lysophosphatidic acid (eg, 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; BB ₂ bombesin receptor; B ₂ bradykinin receptor; Calcium-sensing receptor; KSHV-ORF74 chemokine receptor; NK ₁ 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, which are 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 protein) produces adenosine triphosphate (ATP) through inhibition of adenylate cyclase activity, which converts ATP to cAMP. ) inhibits the production of 3', 5'-cyclic AMP (cAMP).

In some embodiments, the GPCR that interacts with G _iα is 5-hydroxytryptamine receptor (5-HT receptor) types 5-HT ₁ and 5-HT ₅ ; muscarinic acetylcholine receptors such as M ₂ and M ₄ ; adenosine receptors such as A ₁ and A ₃ ; adrenergic receptors such as α _2A , α _2B , and α _2C ; apelin receptor; calcium sensing receptor; cannabinoid receptors CB1 and CB2; Chemokine CXCR4 receptor; dopamine D ₂ , D ₃ , and D ₄ ; GABA _B receptor; such as metabotropic glutamate receptor 2 (mGluR2), metabotropic glutamate receptor 3 (mGluR3), metabotropic glutamate receptor 4 (mGluR4), metabotropic glutamate receptor 6 (mGluR6), metabotropic glutamate receptor 7 (mGluR7), and metabotropic glutamate receptor 8 (mGluR8). glutamate receptor; histamine receptors such as H ₃ and H ₄ 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; Opioid receptors such as δ, κ, μ, and nociceptin receptors; Prostaglandin receptors such as prostaglandin E receptor 1 (EP ₁ ), prostaglandin E receptor 3 (EP ₃ ), prostaglandin F receptor (FP), and thromboxane receptor (TP); 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 encrypted by GNAT3. G _z is encrypted by GNAZ.

G _s family

G _s α (also known as G excitatory, G _s alpha subunit, or G _s protein) is an adenylate conjugate that converts adenosine triphosphate (ATP) to 3',5'-cyclic AMP (cAMP) and pyrophosphate. Activates the cAMP-dependent pathway through activation of clase. In some embodiments, the GPCR that interacts with G _s α is 5-hydroxytryptamine receptor (5-HT receptor) types 5-HT ₄ , 5-HT ₆ , and 5-HT ₇ ; Adrenocorticotropic 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 receptors β ₁ , β ₂ , and β ₃ ; calcitonin receptor; calcitonin gene-related peptide receptor; corticotropin-releasing hormone receptor; Dopamine receptor D1-like family receptors such as D ₁ and D ₅ ; follicle-stimulating hormone receptor (FSH-receptor); gastric inhibitory polypeptide receptor; glucagon receptor; histamine H ₂ receptor; Luteinizing hormone/chorionic gonadotropin receptor; melanocortin receptors such as MC1R, MC2R, MC3R, MC4R, and MC5R; parathyroid hormone receptor 1; Prostaglandin receptor types D ₂ and I ₂ ; secretin receptor; thyrotropin receptor; Trace amine-related receptor 1; and box jelly fish opsin.

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

Additional regulators of the Hippo signaling network

In some embodiments, an 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 contains transmembrane CRB proteins that assemble a multi-protein complex that functions in cell polarity. In some cases, the CRB complex recruits members of the angiomotin (AMOT) family of adapter proteins that interact with Hippo pathway components. In some cases, studies have shown that AMOT binds directly to YAP, promotes YAP phosphorylation and inhibits its nuclear localization.

In some cases, additional regulators of the Hippo signaling pathway include regulators of the MST kinase family. MST kinase monitors actin cytoskeleton integrity. In some cases, the regulators 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) inhibits YAP nuclear localization and activity through regulation of MST activity. In some embodiments, the E-cad-related protein α-catenin regulates YAP by sequestering the YAP/14-3-3 complex in the cytoplasm. In another example, Ajuba protein family members interact with Lats1/2 kinase activity, preventing inactivation of YAP/TAZ.

In some embodiments, additional proteins that directly or indirectly interact with YAP/TAZ include Merlin, protocadherin fat 1, MASK1/2, HIPK2, PTPN14, RASSF, PP2A, salt 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 coactivator/Yes-associated protein transcriptional coactivator (TAZ/YAP) with a PDZ binding motif. In some embodiments, the compounds described herein increase phosphorylation of transcriptional coactivator/Yes-associated protein transcriptional coactivator (TAZ/YAP) with a PDZ binding motif, or increase the phosphorylation of a transcriptional coactivator with a PDZ binding motif. /Reduces dephosphorylation of Yes-Associated Protein Transcriptional Coactivator (TAZ/YAP). In some embodiments, the compound increases ubiquitination of a transcriptional coactivator/Yes-associated protein transcriptional coactivator with a PDZ binding motif (TAZ/YAP), or a transcriptional coactivator/Yes with a PDZ binding motif. -Reduces deubiquitination of transcriptional coactivator-related proteins (TAZ/YAP).

In some embodiments, the compounds disclosed herein are inhibitors of one or more proteins involved in or involved in the Hippo pathway. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of a G-protein and/or a GPCR coupled thereto. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of a G-protein. In some embodiments, inhibitors of the Hippo pathway include G q α family proteins such as G _q , G _q/11 , G _q/14 , and G _{q /15} ; G _12/13 α family proteins such as G ₁₂ and G ₁₃ ; or inhibitors of G _iα family proteins such as 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 _p . 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 G ₁₂ . In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of G ₁₃ . In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of _Gi α1. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of _Gi α2. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of _Gi α3. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of _Gi α4. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of _Go α. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of G _t . 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 G _z .

In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of the core protein of the Hippo pathway. 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 proteins involved in the ubiquitination and proteasome degradation pathways. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of a proteasome degradation pathway protein (e.g., the 26S proteasome).

In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of a protein of the Ras superfamily of proteins. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of a 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, where family members participate in diverse and important cellular processes such as gene transcription, cell-cell adhesion, and cell cycle progression. Cdc42 is involved in cell growth and polarity, and in some cases Cdc42 is activated by guanine nucleotide exchange factor (GEF). In some cases, inhibitors of Cdc42 are compounds disclosed herein.

In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of a deubiquitination enzyme. In some embodiments, the inhibitor of the Hippo pathway is an inhibitor of a cysteine protease or metalloprotease. 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) superfamily of cysteine proteases. In some embodiments, the 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, for treating one or more disorders, diseases and/or conditions, wherein the disorders, diseases or conditions include a cell proliferative disorder. , but is not limited to this.

The activity of compounds used in the invention that are inhibitors of TEADs (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4), or variants or mutants thereof, can be assayed in vitro, in vivo, or in cell lines. there is. In vitro assays include assays that determine inhibition of TEADs (e.g., TEAD1, TEAD2, TEAD3 and/or TEAD4), or variants or mutants thereof. Alternating in vitro assays quantify the ability of an inhibitor to bind to a TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4), or a variant or mutant thereof. Detailed conditions for assaying the compounds used in the invention as inhibitors of TEADs (e.g., TEAD1, TEAD2, TEAD3 and/or TEAD4), or variants or mutants thereof, are set forth in the Examples below. See, for example, Examples 2 and 5.

As used herein, the terms “treatment,” “treat,” and “treating” refer to reversing, alleviating, delaying the onset, or inhibiting the progression of a disease or disorder, or one or more symptoms thereof, as described herein. do. In some embodiments, treatment may be administered after one or more symptoms have occurred. In other embodiments, treatment may be administered before symptoms appear. For example, treatment may be administered to susceptible individuals before symptoms appear (e.g., in light of history of symptoms and/or in light of genetic or other susceptibility factors). Treatment may also be continued even after symptoms have resolved, for example, to prevent or delay recurrence.

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

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 a variant or mutation thereof. means any disease or other adverse condition in which the body is known to play a role. Accordingly, another aspect or embodiment of the invention is to treat or reduce the severity of one or more diseases in which TEADs (e.g., TEAD1, TEAD2, TEAD3 and/or TEAD4), or variants or mutants thereof, are known to play a role. It's about.

As used herein, the term “therapeutically effective amount” refers to reducing the biological activity of a TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4), or a variant or mutant thereof, in a biological sample or in a patient. or attenuating, providing a therapeutic benefit in the treatment of a condition, or delaying or minimizing one or more symptoms associated with the condition. In some embodiments, a “therapeutically effective amount” refers to a measurable binding or signaling activity of a TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4), or a variant or mutant thereof, or any TEAD-mediated activity. Refers to the amount of TEAD inhibitor or pharmaceutically acceptable salt thereof that reduces. The term “therapeutically effective amount” may, in some embodiments, include amounts that improve overall therapy, reduce or avoid symptoms, signs or causes of a condition, and/or enhance the therapeutic efficacy of another therapeutic agent. . In certain embodiments, a therapeutically effective amount is an amount sufficient to inhibit TEAD transcription factors. In certain embodiments, a therapeutically effective amount is an amount sufficient to treat the proliferative disease.

In some aspects and embodiments, an increased TEAD (e.g., TEAD1, TEAD2, Treating a disease or disorder characterized by or associated with TEAD3 and/or TEAD4) expression and/or increased TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4) activity, or one or more symptoms of the disease or disorder Provided herein are methods of reducing severity, delaying onset, or inhibiting progression. In some aspects and embodiments, TEADs (e.g., TEAD1, TEAD2, TEAD3, and /or Provided herein are methods of treating, reducing the severity, delaying the onset, or inhibiting the progression of a disease or disorder, or one or more symptoms of a disease or disorder, in which inhibition or antagonism of TEAD4) activity is beneficial. In some aspects and embodiments, diseases or disorders in which inhibition or antagonism of the Hippo pathway is beneficial, comprising administering a therapeutically effective compound of the invention, or a pharmaceutically acceptable composition thereof, to a patient in need thereof. Provided herein are methods of treating, reducing the severity, delaying the onset, or inhibiting the progression of one or more symptoms of a disease or disorder.

In some aspects and embodiments, the invention includes, but is not limited to, cell proliferative disorders, comprising administering a TEAD inhibitor compound, or pharmaceutical salt or composition thereof, as described herein, to a patient in need thereof. Provided are methods of treating one or more, but not limited to, disorders, diseases and/or conditions. In some embodiments, the cell proliferative disorder is cancer. In some embodiments, the cancer exhibits increased TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4) expression and/or increased TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4) activity. It is characterized by

As used herein, the terms “increased,” “elevated,” or “enhanced” are used interchangeably and include any measurable increase in biological function, and/or biological activity and/or concentration. . For example, an increase in function, activity, or concentration is at least about 10%, 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%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97% , about 98%, about 99%, about 100%, about 2 times, about 3 times, about 4 times, about 5 times, about 6 times, about 7 times, about 8 times, about 9 times, about 10 times, about It can be 20 times, about 25 times, about 50 times, about 100 times or more.

As used herein, the terms “increased expression” and/or “increased activity” of a substance such as a TEAD in a sample or cancer or patient refers to a disease or disorder (as determined by techniques known in the art). e.g., about 5%, about 10%, about 15%, about 20%, about 5%, about 10%, 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%, about 80%, about 85% , about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, about 100%, about 2 times, about 3 times, about 4 times, about 5 times, about 6 times, about It refers to an increase in the amount of a substance such as TEAD by 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. Expression and/or activity of TEAD was 1 standard deviation, 2 standard deviations, 3 standard deviations relative to the mean (average) or median of TEAD in a control group of samples or a baseline group of samples or a retrospective analysis of patient samples. A subject may also be determined to have “increased expression” or “increased activity” of TEAD if it increases by more than 4 standard deviations, 5 standard deviations, or more. As practiced in the art, such control or baseline expression levels can be predetermined, measured, or obtained from a database of such control samples prior to measurement in the sample or cancer or subject.

As used herein, “proliferative disease” refers to a disease that results from abnormal growth or expansion by proliferation of cells (Walker, Cambridge Dictionary of Biology, Cambridge University Press: Cambridge, UK, 1990). Proliferative diseases may be associated with: 1) pathological proliferation of normally quiescent cells; 2) pathological migration of cells from their normal location (e.g., metastasis of neoplastic cells); 3) pathological expression of proteolytic enzymes such as matrix metalloproteinases (e.g. collagenase, gelatinase and elastase); or 4) pathological angiogenesis, such as in proliferative retinopathy and tumor metastases. Exemplary proliferative diseases include cancer (i.e., “malignant neoplasms”), benign neoplasms, angiogenesis, inflammatory diseases, and autoimmune diseases.

cancer

Cancers or proliferative disorders or tumors to be treated using the compounds and methods and uses described herein include hematological cancer, lymphoma, myeloma, leukemia, neurological cancer, skin cancer, breast cancer, prostate cancer, colorectal cancer, lung cancer, head and neck cancer, and gastrointestinal cancer. , liver cancer, pancreatic cancer, genitourinary cancer, bone cancer, kidney cancer, and vascular cancer, but are not limited to these.

In some embodiments of the methods and uses described herein, the cancer is mediated by activation of transcriptional coactivator/Yes-associated protein transcriptional coactivator (TAZ/YAP) with a PDZ binding motif. In some embodiments of the methods and uses described herein, cancer is mediated by modulation of the interaction of YAP/TAZ with TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4). In some embodiments of the methods and uses described herein, the cancer expresses increased TEAD (e.g., TEAD1, TEAD2, TEAD3, and/or TEAD4) and/or has increased TEAD (e.g., TEAD1, 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 the 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 specific percentage of cells in a sample, such as a tumor sample, that have a detectable amount of genetic alteration in one or more Hippo pathway genes. refers to As used herein, genetic alterations in genes, such as Hippo pathway genes, include, for example, loss-of-function mutations in the gene (including, for example, frameshifts, nonsense mutations, and splicing mutations), in gene copy number. (including, for example, copy gain, amplification, copy loss, or deletion), or a fusion of a gene with another gene, for example, a TAZ-CAMTA1 fusion or a YAP1-TFE3 fusion. In some embodiments, the genetic alteration in a Hippo pathway gene occurs in about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35% of cells, such as tumor cells, in the sample. %, 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% have at least about 3 copies of the genetically modified Hippo pathway gene, at least about 4 copies of the genetically modified Hippo pathway gene, At least about 5 copies of a genetically modified Hippo pathway gene, at least about 6 copies of a genetically modified Hippo pathway gene, at least about 7 copies of a genetically modified Hippo pathway gene, at least about about 7 copies of a genetically modified Hippo pathway gene 8 copies, at least about 9 copies of a genetically modified Hippo pathway gene, at least about 10 copies of a genetically modified Hippo pathway gene, at least about 11 copies of a genetically modified Hippo pathway gene, genetically modified At least about 12 copies of a Hippo pathway gene, at least about 9 copies of a genetically modified Hippo pathway gene, at least about 10 copies of a genetically modified Hippo pathway gene, at least about 11 copies of a genetically modified Hippo pathway gene duplicates, at least about 13 copies of a genetically modified Hippo pathway gene, at least about 14 copies of a genetically modified Hippo pathway gene, at least about 15 copies of a genetically modified Hippo pathway gene, at least about 15 copies of a genetically modified Hippo pathway gene refers to having at least about 20 copies of a gene, or more. In some embodiments, a genetic alteration in a Hippo pathway gene refers to such that about 10% of the tumor cells in the sample have at least about 15 copies of a Hippo pathway gene that are genetically altered. In some embodiments, a genetic alteration in a Hippo pathway gene refers to such that about 40% of the tumor cells in the sample have at least about 4 copies of a Hippo pathway gene that are genetically altered. In some embodiments, a genetic alteration in a Hippo pathway gene refers to such that about 10% of the tumor cells in the sample have at least about 4 copies of a Hippo pathway gene that are genetically altered. In some embodiments, the Hippo pathway gene is NF2. In some embodiments, the genetic alteration in one or more Hippo pathway genes is an NF2 deletion. In some embodiments, NF2 deletion refers to an NF2 loss-of-function mutation. In some embodiments, NF2 deletion refers to loss or deletion of NF2 replication. In some embodiments, NF2 deletion refers to no or very low NF2 mRNA expression. In some embodiments, the Hippo pathway gene is YAP1. In some embodiments, the genetic alteration in one or more Hippo pathway genes is YAP1 amplification. In some embodiments, the genetic alteration 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 alteration in one or more Hippo pathway genes is TAZ amplification. In some embodiments, the genetic alteration 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 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 mutant Gα-protein is selected from G12, G13, Gq, G11, Gi, Go, and Gs. In some embodiments, the mutant Gα-protein is G12. In some embodiments, the mutant Gα-protein is G13. In some embodiments, the mutant Gα-protein is Gq. In some embodiments, the mutant Gα-protein is G11. In some embodiments, the mutant Gα-protein is Gi. In some embodiments, the mutant Gα-protein is Go. In some embodiments, the mutant Gα-protein is Gs.

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

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, 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, at least about 3 years, at least It represents progression-free survival of 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, 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, at least about 18 months, at least Indicates an overall survival of 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, patients treated using the methods or uses described herein have at least about 15%, at least about 20%, at least about 25%, at least about 30%, about 35%, about 40%, about 45%. , an objective response rate of about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%. response rate (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 mesothelioma. In some embodiments, the cancer is lung cancer, thyroid cancer, ovarian cancer, colorectal cancer, prostate cancer, pancreatic cancer, esophageal cancer, liver cancer, breast cancer, skin cancer, or mesothelioma, sarcoma, or epithelioid hemangioendothelioma (EHE). In some embodiments, the cancer is mesothelioma, such as malignant mesothelioma. In some embodiments, the cancer is EHE.

In some embodiments, the cancer includes, but is not limited to, leukemia (e.g., acute leukemia, acute lymphoblastic leukemia, acute myelocytic leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myelocytic leukemia, chronic lymphocytic leukemia), polycythemia vera, lymphoma (e.g., Hodgkin's disease or non-Hodgkin's disease), Waldenstrom's macroglobulinemia, multiple myeloma, heavy chain disease, and Solid tumors such as sarcomas and carcinomas (e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendothelioma, synovium, 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 carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchial carcinoma. , renal cell carcinoma, liver tumor, cholangiocarcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterine cancer, testicular cancer, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, etc. Glioblastoma type (GBM, also known as glioblastoma), medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma, melanoma, neuroblastoma, and retinoblastoma) Includes.

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

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

In another embodiment, the cancer includes, but is not limited to, mesothelioma, hepatobiliary (liver and bile duct), bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or intraocular melanoma, ovarian cancer, colon cancer, rectal cancer, anal cancer, stomach cancer, gastrointestinal cancer. Cancer (stomach, colorectal and duodenum), uterine cancer, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, 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 myelogenous leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, non-Hodgkin's lymphoma, spinal cord tumor, brainstem neuroblastoma. glioma, pituitary adenoma, adrenocortical cancer, gallbladder cancer, multiple myeloma, cholangiocarcinoma, fibrosarcoma, neuroblastoma, retinoblastoma, or a combination of one or more of the foregoing.

In some embodiments, the cancer is hepatocellular carcinoma, ovarian cancer, ovarian epithelial cancer, or fallopian tube cancer; Papillary serous cystadenocarcinoma or uterine papillary serous carcinoma (UPSC); prostate cancer; testicular cancer; Gallbladder cancer; hepatobiliary carcinoma; Soft tissue and bone synovial sarcoma; rhabdomyosarcoma; osteosarcoma; chondrosarcoma; Ewing's sarcoma; Anaplastic thyroid cancer; Adrenocortical adenoma; pancreatic cancer; pancreatic ductal carcinoma 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 associated malignant peripheral nerve sheath tumor (MPNST); Waldenstrom's macroglobulinemia; or medulloblastoma.

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

In some embodiments, the cancer is a solid tumor, such as a sarcoma, carcinoma, or lymphoma. Solid tumors typically contain abnormal masses of tissue that do not typically 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; rectal cancer; anal cancer; lung cancer, such as non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC); Ovarian cancer, ovarian epithelial cancer, ovarian carcinoma, or fallopian tube cancer; Papillary serous cystadenocarcinoma or uterine papillary serous carcinoma (UPSC); prostate cancer; testicular cancer; Gallbladder cancer; hepatobiliary carcinoma; Soft tissue and bone synovial sarcoma; rhabdomyosarcoma; osteosarcoma; chondrosarcoma; Ewing's sarcoma; Anaplastic thyroid cancer; Adrenocortical carcinoma; pancreatic cancer; pancreatic ductal carcinoma 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 associated malignant peripheral nerve sheath tumor (MPNST); Waldenstrom's macroglobulinemia; or medulloblastoma.

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, ovarian epithelial cancer, ovarian carcinoma, fallopian tube cancer, papillary serous Cystadenocarcinoma, uterine papillary serous carcinoma (UPSC), hepatobiliary carcinoma, soft tissue and bone synovial sarcoma, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, anaplastic thyroid cancer, adrenocortical carcinoma, pancreatic cancer, pancreatic ductal carcinoma, pancreatic adenocarcinoma, glioma. , brain cancer, neurofibromatosis-1 associated malignant peripheral nerve sheath tumor (MPNST), Waldenstrom's macroglobulinemia, or medulloblastoma.

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

In some embodiments, the cancer is caused by human T-cell leukemia virus type I (HTLV-I) and is characterized by human immunodeficiency, a highly aggressive form of CD4+ T cell leukemia characterized by clonal integration of HTLV-I in leukemic cells. Virus-related cancers, including HIV-related solid tumors, human papillomavirus (HPV)-16 positive refractory solid tumors, and adult T-cell leukemia (https://clinicaltrials.gov/ct2/show/study/NCT02631746) as well as virus-related tumors in gastric cancer, nasopharyngeal carcinoma, 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; See also 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 according to the methods of the present invention can be administered using any amount and any route of administration effective for treating or lessening the severity of cancer or tumor. The exact amount required will vary from subject to subject depending on the subject's species, age and general condition, the severity of the disease or condition, the particular agent, its mode of administration, etc. The compounds of the present invention are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. As used herein, the expression “dosage unit form” refers to a physically discrete unit of agent suitable for the patient to be treated. However, it will be understood that the total daily usage amount of the compounds and compositions of the present invention will be determined 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 a variety of factors, including the disorder being treated and the severity of the disorder; the activity of the specific compound used; the specific composition used; The patient's age, weight, general health, gender and diet; time of administration, route of administration, and/or rate of excretion of the particular compound used; duration of treatment; Drugs used in conjunction with or simultaneously with the specific compound used; and similar factors well known in the medical 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 present invention can be administered to humans and other animals orally, rectally, parenterally, intracisternally, vaginally, intraperitoneally, or topically (as a powder), depending on the severity of the disease or disorder being treated. , as an ointment or drops), bucally, orally or as a nasal spray, etc. In certain embodiments, the compounds of the invention are administered to the subject at about 0.01 mg/kg to about 50 mg/kg, and preferably at about 1 mg/kg to about 25 mg/kg, at least once per day to achieve the desired therapeutic effect. It may be administered orally or parenterally at dosage levels of body weight/day.

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 can be mixed, for example, with water or other solvents, solubilizers and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3- Butylene glycol, dimethylformamide, oils (especially cotton oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan. , and mixtures thereof. In addition to inert diluents, oral compositions may also 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 the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a non-toxic, parenterally acceptable diluent or solvent, for example, a solution in 1,3-butane diol. Among the acceptable vehicles and solvents that may be used are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. Additionally, sterile, fixed oils are commonly used as solvents or suspending media. For this purpose any mild fixed oil may be used including synthetic mono or diglycerides. Additionally, fatty acids such as oleic acid are used in the preparation of injectables.

Injectable formulations may be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use. there is.

To prolong the effect of the compounds of the invention, it is usually desirable to slow the absorption of the compounds from subcutaneous or intramuscular injection. This can be accomplished using liquid suspensions of crystalline or amorphous materials with low water solubility. The rate of absorption of the compound then depends on its dissolution rate, which in turn may depend on crystal size and crystal shape. Alternatively, delayed absorption of a parenterally administered compound form is achieved by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming a microcapsule matrix of the compound in a biodegradable polymer such as polylactide-polyglycolide. Depending on the ratio of compound to polymer and the nature of the particular polymer used, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Injectable depot formulations are also prepared by entrapment of compounds in liposomes or microemulsions that are compatible with body tissues.

Compositions for rectal or vaginal administration preferably contain a suitable non-irritating excipient, such as cocoa butter, polyethylene glycol or suppository wax, which is solid at ambient temperature but liquid at body temperature and melts in the rectal or vaginal cavity to release the active compound. It is a suppository that can be manufactured by mixing with a carrier.

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

Solid compositions of a similar type can also be used as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol, etc. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared using coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulation art. These may optionally contain opacifying agents and may be compositions that release the active ingredient(s) alone, preferentially 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. Solid compositions of a similar type can also be used as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols, etc.

The active compounds may also be in micro-encapsulated form with one or more excipients as mentioned above. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared using coatings and shells such as enteric coatings, controlled release coatings and other coatings well known in the pharmaceutical formulation art. In these solid dosage forms, the active compound may be mixed with at least one inert diluent such as sucrose, lactose or starch. These dosage forms may also contain additional substances other than inert diluents, such as tabletting lubricants, and other tableting aids such as magnesium stearate and microcrystalline cellulose, as is common practice. For capsules, tablets and pills, the dosage form may also include buffering agents. These may optionally contain opacifying agents and may also be compositions that release the active ingredient(s) only, or preferentially, 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 the invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any necessary preservatives or buffers that may be required. Ophthalmic formulations, ear drops, and eye drops are also considered to be within the scope of the present invention. Additionally, the present invention contemplates the use of transdermal patches, which have the additional advantage of providing controlled delivery of the compound to the body. These dosage forms can be made by dissolving or dispensing the compound in an appropriate medium. Absorption enhancers can also be used to increase the flux of compounds across the skin. The rate can be controlled by providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.

Concomitant administration with one or more other therapeutic agent(s)

Depending on the particular condition or disease being treated, additional therapeutic agents that are typically administered to treat that condition may also be present in the compositions of the invention. As used herein, additional therapeutic agents that are generally administered to treat a particular disease or condition are said to be "appropriate for the disease or condition being treated."

In some embodiments, the invention provides an effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof, administered to a patient in need thereof, and an effective amount of one or more additional therapeutic agents, such as those described herein, simultaneously or sequentially. Provided is a method of treating the disclosed disease or condition, comprising co-administration. In some embodiments, the method comprises co-administering one additional therapeutic agent. In some embodiments, the method comprises coadministering two additional therapeutic agents. In some embodiments, combinations of disclosed compounds with an additional therapeutic agent or therapeutic agents act synergistically.

The compounds of the invention may also be used in conjunction with known therapeutic procedures, such as administration of hormones or radiation. In certain embodiments, provided compounds are used as radiosensitizers, particularly for the treatment of tumors that exhibit poor sensitivity to radiotherapy.

The compounds of the present invention may be administered alone or in combination with one or more other therapeutic compounds, and possible combination therapy may take the form of fixed combinations, or combined administration of a fixed combination with one or more other therapeutic compounds, or of the present invention. Administration of the compound and one or more other therapeutic compounds may be staggered or provided independently of one another. The compounds of the invention can be administered separately or additionally for tumor treatment, especially in combination with chemotherapy, radiotherapy, immunotherapy, phototherapy, surgical intervention, or combinations thereof. As explained above, long-term therapy is equally possible as is adjuvant therapy in the context of other treatment strategies. Other possible treatments are therapy to maintain the patient's condition after tumor regression, or even chemotherapy, for example, in patients at risk.

One or more other therapeutic agent(s) may be administered separately from the compound or composition of the invention as part of a multiple dose regimen. Alternatively, one or more other therapeutic agent(s) may be part of a single dosage form, mixed together with the compound of the invention in a single composition. When administered in a multiple dose regimen, one or more other therapeutic agent(s) and a compound or composition of the invention may be administered simultaneously, sequentially or within a time period of each other, e.g., 1, 2, 3, 4, 5, 6 from each other. , 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 18, 20, 21, 22, 23, or 24 hours. In some embodiments, one or more other therapeutic agent(s) and a compound or composition of the invention are administered in a multiple dose regimen within an interval of more than 24 hours.

As used herein, the terms “combination,” “combined,” and related terms refer to simultaneous or sequential administration of therapeutic agents according to the invention. For example, the compounds of the invention can be administered in separate unit dosage forms or in single unit dosage form together with one or more other therapeutic agent(s) simultaneously or sequentially. Accordingly, the present 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.

The amounts of a compound of the invention and one or more other therapeutic agent(s) (in such compositions comprising additional therapeutic agents as described above) that can be combined with a carrier material to produce a single dosage form will vary depending on the host treated and the particular mode of administration. It will vary depending on. Preferably, the composition of the present invention should be formulated so that a dosage of 0.01 - 100 mg/kg body weight/day of the compound of the present invention can be administered.

In such compositions comprising 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 the amount required in monotherapy using that therapeutic agent alone. In such compositions, dosages of 0.01 - 1,000 μg/kg body weight/day of one or more other therapeutic agent(s) may be administered.

The amount of one or more other therapeutic agents present in the compositions of the present invention may be less 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 other therapeutic agent(s) in the presently disclosed compositions will range from about 50% to 100% of the amount normally present in a composition comprising that therapeutic agent as the only therapeutically active agent. In some embodiments, one or more other therapeutic agent(s) is administered in about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, or about 80% of the amount typically administered for that therapeutic agent. , administered at a dosage of about 85%, about 90%, or about 95%. As used herein, the phrase “normally administered” refers to the amount of an FDA-approved therapeutic agent approved for administration, per FDA label insert.

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

Exemplary Other Remedies

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®, Spectrum Pharmaceuticals); Entinostat (SNDX-275, Syndax Pharmaceuticals) (NCT00866333); and chidamide (EPIDAZA®, HBI-8000, Chipscreen Biosciences, China).

In some embodiments, the one or more other therapeutic agents are CDK inhibitors, such as CDK4/CDK6 inhibitors. In some embodiments, the CDK 4/6 inhibitor is palbociclib (IBRANCE®, Pfizer); ribociclib (KISQALI®, Novartis); Abemaciclib (Ly2835219, Eli Lilly); and trilaciclib (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); Copanlisib (BAY806946, Bayer); duvelisib (formerly IPI-145, Infinity Pharmaceuticals); PQR309 (Piqur Therapeutics, Switzerland); and TGR1202 (formerly RP5230, TG Therapeutics).

In some embodiments, the one or more other therapeutic agents are platinum-based therapeutic agents, also referred to as platin. Platin causes cross-linking of DNA, primarily in rapidly reproducing cells, such as cancer cells, thereby inhibiting DNA repair and/or DNA synthesis.

In some embodiments, the platinum-based therapeutic agent is cisplatin (PLATINOL®, Bristol-Myers Squibb); carboplatin (PARAPLATIN®, Bristol-Myers Squibb; also Teva; Pfizer); Oxaliplatin (ELOXITIN® Sanofi-Aventis); nedaplatin (AQUPLA®, Shionogi), picoplatin (Poniard Pharmaceuticals); and satraplatin (JM-216, Agennix).

In some embodiments, one or more other therapeutic agents are taxane compounds that cause destruction of microtubules essential for cell division. In some embodiments, the taxane compound is paclitaxel (TAXOL®, Bristol-Myers Squibb), docetaxel (TAXOTERE®, Sanofi-Aventis; DOCEFREZ®, Sun Pharmaceutical), albumin-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 therapeutic agents that will interfere with normal DNA synthesis, protein synthesis, cell replication, or otherwise inhibit rapidly proliferating cells.

In some embodiments, the nucleoside inhibitor is trabectedin (guanidine alkylating agent, YONDELIS®, Janssen Oncology), mechlorethamine (alkylating agent, VALCHLOR®, Aktelion Pharmaceuticals); Vincristine (ONCOVIN®, Eli Lilly; VINCASAR®, Teva Pharmaceuticals; MARQIBO®, Talon Therapeutics); Temozolomide (TEMODAR®, Merck, a prodrug for the alkylating agent 5-(3-methyltriazen-1-yl)-imidazole-4-carboxamide (MTIC)); cytarabine injection (ara-C, an anti-metabolic cytidine analog, Pfizer); Lomustine (alkylating agent, CEENU®, Bristol-Myers Squibb; GLEOSTINE®, NextSource Biotechnology); Azacytidine (pyrimidine nucleoside analog of cytidine, VIDAZA®, Celgene); Omacetaxin mephesuccinate (cephalotaxin ester) (protein synthesis inhibitor, SYNRIBO®; Teva Pharmaceuticals); Asparaginase Erwinia chrysanthemi (asparagine depleting enzyme, ELSPAR®, Lundbeck; ERWINAZE®, EUSA Pharma); Eribulin mesylate (microtubule inhibitor, tubulin-based antimitotic agent, HALAVEN®, Eisai); cabazitaxel (microtubule inhibitor, tubulin-based antimitotic agent, JEVTANA®, Sanofi-Aventis); capacetrin (thymidylate synthase inhibitor, XELODA®, Genentech); Bendamustine (bifunctional mechlorethamine derivative, believed to form interstrand DNA cross-links, TREANDA®, Cephalon/Teva); Ixabepilone (semisynthetic analog of epothilone B, microtubule inhibitor, tubulin-based antimitotic agent, IXEMPRA®, Bristol-Myers Squibb); nelarabine (prodrug of a deoxyguanosine analogue, inhibitor of nucleoside metabolism, ARRANON®, Novartis); Chlorapavine (prodrug of ribonucleotide reductase inhibitor, competitive inhibitor of deoxycytidine, CLOLAR®, Sanofi-Aventis); and trifluridine and tipiracil (thymidine-based nucleoside analog and thymidine phosphorylase inhibitor, LONSURF®, Taiho Oncology).

In some embodiments, the one or more other therapeutic agents are a kinase inhibitor or VEGF-R antagonist. In some embodiments, the one or more other therapeutic agents are MEK inhibitors. As used herein, “MEK inhibitor” refers to any inhibitor or blocker or antagonist that binds to and/or inhibits the mitogen activated protein kinase enzymes MEK1 and/or MEK2. In some embodiments, the MEK inhibitor is selected from those described in Cheng et al., “Current Development Status of MEK Inhibitors,” Molecules 2017 , 22, 1551, the entire contents of which are incorporated herein by reference. In certain embodiments, the MEK inhibitor is binimetinib (MEK162, ARRY-438162, ARRAY BIOPHARMA INC.), cobimetinib (COTELLIC®, Exelexis/Genentech/ROChe), lepametinib (BAY 86-9766, RDEA119; Bayer AG), selumetinib (AZD6244, ARRY-142886; ASTRAZENECA), trametinib (MEKINIST®, Novartis), mirdametinib (PD-0325901, Spring Works Therapeutics), pimasertib (AS703026, MSC1936369B, Merck KGaA) or pharmaceutically acceptable salts and/or solvates of any of the foregoing. In certain embodiments, the second anti-cancer agent is binimetinib, cobimetinib, selumetinib, trametinib, mirdametinib, pimasertib, or a pharmaceutically acceptable salt and/or solvent of any of the foregoing. It's cargo. Other examples of MEK inhibitors for use as other therapeutic agents in the methods and uses described herein include E6201 (Eisai Co Ltd./Strategia Theraputics), GDC-0623 (RG 7421, Genentech, Inc.), CH5126766 (RO5126766, Chugai 232 Pharmaceuticals) 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 anticancer agents in the methods and uses described herein include WO2005/121142, WO2014/169843, WO2016/035008, WO2016/168704, WO2020/125747, WO2021/142144, WO2021/142345, WO2021 The entire contents of each of these patent documents, including but not limited to those described in /149776, are incorporated herein by reference.

In some embodiments, the one or more other therapeutic agents are EGFR inhibitors. As used herein, “EGFR inhibitor” refers to any inhibitor or blocker or antagonist that binds to and/or inhibits the epidermal growth factor receptor (EGFR). In some embodiments, the EGFR inhibitor is selected from those described in Ayati et al., "A review on progression of epidermal growth factor receptor (EGFR) inhibitors as an efficient approach in cancer targeted therapy," Bioorganic Chemistry 2020 , 99: 103811, The entire contents of the above document are incorporated herein by reference. In some embodiments, the EGFR inhibitor is selected from cetuximab, necitumumab, panitumumab, zalutumumab, 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 zalutumumab. 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 TKIs refer to reversible EGFR inhibitors, such as gefitinib and erlotinib, which are effective in the first-line treatment of NSCLC with EGFR activating mutations, such as deletions in exon 19 and exon 21 L858R mutation.

In some embodiments, the EGFR inhibitor is a “second generation EGFR tyrosine kinase inhibitor” (second generation TKI). Second generation TKIs refer to covalent irreversible EGFR inhibitors, such as afatinib and dacomitib, which are effective in the first-line treatment of NSCLC with EGFR activating mutations, such as deletions in exon 19 and exon 21 L858R mutation.

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

In some embodiments, one or more other therapeutic agents are selected from approved VEGF inhibitors and kinase inhibitors useful in the invention, including: bevacizumab (AVASTIN®, Genentech/ROChe), an anti-VEGF monoclonal antibody; Ramucirumab (CYRAMZA®, Eli Lilly), an anti-VEGFR-2 antibody, and Zip-aflibercept (ZALTRAP®; Regeneron/Sanofi), also known as the VEGF Trap. 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 such as cobimetanib (COTELLIC®, Exelexis/Genentech/ROChe); trametinib (MEKINIST®, Novartis); Bcr-Abl tyrosine kinase inhibitors such as imatinib (GLEEVEC®, Novartis); nilotinib (TASIGNA®, Novartis); dasatinib (SPRYCEL®, BristolMyersSquibb); bosutinib (BOSULIF®, Pfizer); and ponatinib (INCLUSIG®, Ariad Pharmaceuticals); Her2 and EGFR inhibitors such as gefitinib (IRESSA®, AstraZeneca); erlotinib (TARCEEVA®, Genentech/ROChe/Astellas); lapatinib (TYKERB®, Novartis); Afatinib (GILOTRIF®, Boehringer Ingelheim); Osimertinib (targets activated EGFR, TAGRISSO®, AstraZeneca); and brigatinib (ALUNBRIG®, Ariad Pharmaceuticals); c-Met and VEGFR2 inhibitors such as cabozantinib (COMETRIQ®, Exelexis); and multikinase inhibitors such as sunitinib (SUTENT®, Pfizer); pazopanib (VOTRIENT®, Novartis); ALK inhibitors such as crizotinib (XALKORI®, Pfizer); ceritinib (ZYKADIA®, Novartis); and alectinib (ALECENZa®, Genentech/ROChe); Bruton's tyrosine kinase inhibitors such as ibrutinib (IMBRUVICA®, Pharmacyclics/Janssen); and Flt3 receptor inhibitors such as midostaurin (RYDAPT®, Novartis).

Other kinase inhibitors and VEGF-R antagonists that are in development and that may be used in the present invention include tivozanib (Aveo Pharmaecuticals); Batalanib (Bayer/Novartis); lusitanib (Clovis Oncology); Dovitinib (TKI258, Novartis); Ciauanib (Chipscreen Biosciences); CEP-11981 (Cephalon); linipanib (Abbott Laboratories); neratinib (HKI-272, Puma Biotechnology); radotinib (SUPECT®, IY5511, Il-Yang Pharmaceuticals, S. Korea); ruxolitinib (JAKAFI®, Incyte Corporation); PTC299 (PTC Therapeutics); CP-547,632 (Pfizer); foretinib (Exelexis, GlaxoSmithKline); Includes quizatinib (Daiichi Sankyo) and motesanib (Amgen/Takeda).

In some embodiments, one or more other therapeutic agents are mTOR inhibitors that inhibit cell proliferation, angiogenesis, and glucose uptake. In some embodiments, the mTOR inhibitor is 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).

In some embodiments, the one or more other therapeutic agents are growth factor antagonists, such as platelet-derived growth factor (PDGF), or an antagonist 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 (targeting activated EGFR, TAGRISSO®, AstraZeneca).

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

In some embodiments, the one or more other therapeutic agents are antagonists of the hedgehog pathway. Approved Hedgehog pathway inhibitors that may be used in the present invention include sonidegib (ODOMZO®, Sun Pharmaceuticals), both for the treatment of basal cell carcinoma; and vismodegib (ERIVEDGE®, 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 under investigation that may be 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 under investigation that may be used in the present invention include AG120 (Celgene; NCT02677922); AG221 (Celgene, NCT02677922; NCT02577406); BAY1436032 (Bayer, NCT02746081); Includes IDH305 (Novartis, NCT02987010).

In some embodiments, the one or more other therapeutic agents are arginase inhibitors. An investigational arginase inhibitor that may be used in the present invention is AEB1102 (a pegylated recombinant arginase, Aeglea Biotherapeutics ); and CB-1158 (Calithera Biosciences).

In one embodiment, the one or more other therapeutic agents are glutaminase inhibitors. Glutaminase inhibitors under investigation that may 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, i.e., proteins expressed on the cell surface of tumor cells. Approved antibodies that bind tumor antigens that may 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 (anti-glycolipid GD2, UNITUXIN®, United Therapeutics); Trastuzumab (anti-HER2, HERCEPTIN®, Genentech); Ado-trastuzumab emtansine (anti-HER2 fused to emtansine, KADCYLA®, Genentech); and Pertuzumab (anti-HER2, PERJETA®, Genentech); and brentuximab vedotin (anti-CD30-drug conjugate, ADCETRIS®, Seattle Genetics).

In some embodiments, one or more other therapeutic agents are topoisomerase inhibitors. Approved topoisomerase inhibitors useful in the present invention include irinotecan (ONIVYDE®, Merrimack Pharmaceuticals); Includes topotecan (HYCAMTIN®, GlaxoSmithKline). Topoisomerase inhibitors under investigation that may be used 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 agents that can be used in the present invention include VENCLEXTA® (AbbVie/Genentech); and blinatumomab (BLINCYTO®, Amgen). Other therapeutic agents targeting anti-apoptotic proteins that have been tested in clinical trials and may 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/Medivation); Approved androgen synthesis inhibitors include abiraterone (ZYTIGA®, Centocor/Ortho); Contains an approved antagonist of the gonadotropin-releasing hormone (GnRH) receptor (Degaralix, FIRMAGON®, Ferring Pharmaceuticals).

In some embodiments, one or more other therapeutic agents are selective estrogen receptor modulators (SERMs) that interfere with the synthesis or activity of estrogen. 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. Approved therapeutics that inhibit bone resorption bind to RANKL, preventing its binding to its receptor RANK, its precursor, and osteoclast-like giant cells found on the surface of osteoclasts, thereby preventing solid tumors with bone metastases. denosumab (XGEVA®, Amgen), an antibody that mediates bone pathology. Other approved treatments 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 between the two primary p53 inhibitory proteins, MDMX and MDM2. Inhibitors of p53 inhibitory proteins under investigation that may be used in the present invention include ALRN-6924 (Aileron), a staple peptide that binds equally to MDMX and MDM2 and interferes with the interaction of p53 with MDMX and MDM2. ALRN-6924 is currently being evaluated in clinical trials for the treatment of AML, advanced myelodysplastic syndrome (MDS), and peripheral T-cell lymphoma (PTCL) (NCT02909972; NCT02264613).

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

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

In some embodiments, the one or more other therapeutic agents are antiproliferative compounds. These antiproliferative compounds include aromatase inhibitors; antiestrogens; Topoisomerase I inhibitor; Topoisomerase II inhibitor; microtubule-activating compounds; alkylated compounds; histone deacetylase inhibitors; Compounds that induce cell differentiation processes; cyclooxygenase inhibitors; MMP inhibitors; mTOR inhibitor; anti-tumor anti-metabolites; platinum compounds; Compounds that target/reduce protein or lipid kinase activity and additional anti-angiogenic compounds; Compounds that target, reduce or inhibit the activity of protein or lipid phosphatases; gonadorelin agonist; anti-androgen; methionine aminopeptidase inhibitor; Matrix metalloproteinase inhibitors; bisphosphonate; biological response modifier; antiproliferative antibodies; heparanase inhibitor; Inhibitors of Ras oncogenic isoforms; telomerase inhibitor; proteasome inhibitors; Compounds used in the treatment of blood cancer; Compounds that target, reduce or inhibit the activity of Flt-3; Hsp90 inhibitors such as 17-AAG (17-allylaminogeldanamycin, NSC330507), 17-DMAG (17-dimethylaminoethylamino-17-demethoxy-geldanamycin, NSC707545), IPI-504, CNF1010 from Conforma Therapeutics , CNF2024, CNF1010; temozolomide (TEMODAL ^® ); Kinesin spindle protein inhibitors such as SB715992 or SB743921 from GlaxoSmithKline, or pentamidine/chlorpromazine from CombinatoRx; MEK inhibitors include, but are not limited to, ARRY142886 from Array BioPharma, AZd ₆ 244 from AstraZeneca, PD181461 from Pfizer, and leucovorin.

As used herein, the term “aromatase inhibitor” refers to a compound that inhibits estrogen production, e.g., the conversion of the substrates androstenedione and testosterone to estrone and estradiol, respectively. The term refers to steroids, especially atamestane, exemestane and formestane, and especially non-steroids, especially aminoglutethimide, rogletimide, pyridoglutethimide, trilostane, testolactone, ketoconazole, boro Includes, but is not limited to, zole, fadrozole, anastrozole, and letrozole. Exemestane is marketed under the brand name AROMASIN™. Formestane is marketed under the trade name LENTARON™. Fadrozole is marketed under the brand name AFEMA™. Anastrozole is marketed under the brand name ARIMIDEX™. Letrozole is marketed under the trade names FEMARA™ or FEMAr™. Aminoglutethimide is marketed under the brand name ORIMETEN™. Combinations of the invention comprising a chemotherapeutic agent that is an aromatase inhibitor are particularly useful in the treatment of hormone receptor positive tumors, such as breast tumors.

As used herein, the term “antiestrogens” refers to compounds that antagonize the effects of estrogen at the estrogen receptor level. The term includes, but is not limited to, tamoxifen, fulvestrant, raloxifene and raloxifene hydrochloride. Tamoxifen is marketed under the brand name NOLVADEX™. Raloxifene hydrochloride is marketed under the trade name EVISTA™. Fulvestrant may be administered under the brand name FASLODEX™. Fulvestrant may be administered under the brand name FASLODEX™. Combinations of the invention comprising a chemotherapy agent that is an antiestrogen 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 that can inhibit the biological effects of androgen hormones, including but not limited to bicalutamide (CASODEX™). As used herein, the term “gonadorelin agonist” includes, but is not limited to, abarelix, goserelin, and goserelin acetate. Goserelin may be administered under the brand name ZOLADEX™.

As used herein, the term "topoisomerase I inhibitor" refers to topotecan, gimatecan, irinotecan, camptothecan and its analogs, 9-nitrocamptothecin and the macromolecular camptothecin conjugate PNU- Includes, but is not limited to, 166148. Irinotecan can be administered, eg, in the form as it is commercially available, eg under the brand name CAMPTOSAR™. Topotecan is marketed under the brand 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, anthraquinones. Includes, but is not limited to, mitoxantrone and rosoxantrone, and the podophyllotoxins etoposide and teniposide. Etoposide is marketed under the brand name ETOPOPHOS™. Teniposide is marketed under the trade name VM 26-Bristol, and doxorubicin is marketed under the trade names ACRIBLASTIN™ or ADRIAMYCIN™. Epirubicin is marketed under the brand name FARMORUBICIN™. Idarubicin is marketed under the brand name ZAVEDOS™. Mitoxantrone is marketed under the brand name NOVANTRON™.

The term “microtubule activator” includes taxanes such as paclitaxel and docetaxel; Vinca alkaloids such as vinblastine or vinblastine sulfate, vincristine or vincristine sulfate, and vinorelbine; discordermolide; It relates to microtubule stabilizing, microtubule destabilizing compounds and microtubule polymerization inhibitors, including but not limited to coccicin and epothilone and their derivatives. Paclitaxel is marketed under the brand name TAXOL™. Docetaxel is marketed under the brand name TAXOTERE™. Vinblastine sulfate is marketed under the brand name VINBLASTIN R.P™. Vincristine sulfate is marketed 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 Gliadel). Cyclophosphamide is marketed under the trade name CYCLOSTIN™. Ifosfamide is marketed under the brand name HOLOXAN™.

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

The term “antitumor antimetabolite” refers to DNA demethylating compounds such as 5-fluorouracil or 5-FU, capecitabine, gemcitabine, 5-azacytidine and decitabine, methotrexate and edatrexate, and pemetrexate. These include, but are not limited to, folic acid antagonists such as seeds. Capecitabine is marketed under the brand name XELODA™. Gemcitabine is marketed under the brand name GEMZAR™.

As used herein, the term “platin compounds” includes, but is not limited to, carboplatin, cis-platin, cisplatinum, and oxaliplatin. Carboplatin can be administered, for example, in the form as it is commercially available, e.g. under the brand name CARBOPLAT™. Oxaliplatin can be administered, for example, in the form as it is commercially available, e.g. under the brand name ELOXATIN™.

As used herein, the term "protein or lipid kinase activity; or a compound that targets and/or reduces protein or lipid phosphatase activity, or an additional anti-angiogenic compound" refers to protein tyrosine kinase and/or serine and/or Threonine kinase inhibitors or lipid kinase inhibitors, such as a) compounds that target, reduce or inhibit the activity of platelet-derived growth factor-receptor (PDGFR), such as compounds that target, reduce or inhibit the activity of PDGFR, Compounds that particularly inhibit the PDGFR receptor, such as N-phenyl-2-pyrimidin-amine derivatives such as imatinib, SU101, SU6668 and GFB-111; b) compounds that target, reduce or inhibit the activity of fibroblast growth factor-receptor (FGFR); c) Compounds targeting, reducing or inhibiting the activity of insulin-like growth factor receptor I (IGF-IR), such as compounds targeting, reducing or inhibiting the activity of IGF-IR, especially the IGF-I receptor Compounds that inhibit the kinase activity of, or antibodies targeting the extracellular domain of the IGF-I receptor or growth factor thereof; d) compounds that target, reduce or inhibit the activity of the Trk receptor tyrosine kinase family, or ephrin B4 inhibitors; e) compounds targeting, reducing or inhibiting the activity of the AxI receptor tyrosine kinase family; f) compounds that target, reduce or inhibit the activity of Ret receptor tyrosine kinase; g) compounds that target, reduce or inhibit the activity of the Kit/SCFR receptor tyrosine kinase, such as imatinib; h) Compounds that target, reduce or inhibit the activity of the C-kit receptor tyrosine kinase, which is part of the PDGFR family, such as compounds that target, reduce or inhibit the activity of the c-Kit receptor tyrosine kinase family, especially c -Compounds that inhibit Kit receptors, such as imatinib; i) Compounds that target and reduce or inhibit the activity of c-Abl family members, their gene fusion products (e.g., BCR-Abl kinase) and mutants, such as c-Abl family members and their gene fusions Compounds that target and reduce or inhibit the activity of the product, such as N-phenyl-2-pyrimidin-amine derivatives such as imatinib or nilotinib (AMN107); PD180970; AG957; NSC 680410; PD173955 by ParkeDavis; or dasatinib (BMS-354825); j) Protein Kinase C (PKC) and members of the Raf family of serine/threonine kinases, MEK, SRC, JAK/pan-JAK, FAK, PDK1, PKB/Akt, Ras/MAPK, PI3K, SYK, TYK2, BTK and TEC. Compounds that target, reduce or inhibit the activity of members of the cyclin dependent kinase family (CDK), including members of the family and/or staurosporine derivatives such as midostaurin; Examples of additional compounds include UCN-01, safingol, BAY 43-9006, bryostatin 1, perifosine; Ilmofosin; RO 318220 and RO 320432; GO 6976; Isis 3521; LY333531/LY379196; isokinoline compounds; FTI; Contains PD184352 or QAN697 (P13K inhibitor) or AT7519 (CDK inhibitor); k) Compounds that target, reduce or inhibit the activity of protein-tyrosine kinase inhibitors, such as imatinib mesylate (GLEEVEC™) or Lephostin, such as Tyrphostin A23/RG-50810; AG 99; Tyrphostin AG 213; Tyrpostin AG 1748; Tyrphostin AG 490; Tyrphostin B44; Tyrphostin B44(+) enantiomer; Tyrphostin AG 555; AG494; Tyrphostin AG 556, AG957 and adaphostin (4-{[(2,5-dihydroxyphenyl)methyl]amino}-benzoic acid adamantyl ester; NSC 680410, including adaphostin); l) Compounds targeting, reducing or inhibiting the activity of the epidermal growth factor family of receptor tyrosine kinases (EGFR ₁ , ErbB2, ErbB3, ErbB4 as homodimers or heterodimers) and their mutants, such as epidermal growth Compounds that target, reduce or inhibit the activity of the factor receptor family include, in particular, members of the EGF receptor tyrosine kinase family, such as compounds, proteins or compounds that inhibit EGF receptors, ErbB2, ErbB3 and ErbB4, or bind to EGF or EGF-related ligands. Antibodies, 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 the c-Met receptor, such as compounds that target, reduce or inhibit the activity of the c-Met receptor, especially compounds that inhibit the kinase activity of the c-Met receptor , or antibodies targeting the extracellular domain of c-Met or binding to HGF, n) PRT-062070, SB-1578, baricitinib, pacritinib, momelotinib, VX-509, AZD-1480, TG -101348, targeting and reducing the kinase activity of one or more JAK family members (JAK1/JAK2/JAK3/TYK2 and/or pan-JAK), including but not limited to tofacitinib, and ruxolitinib. Compounds that induce or inhibit; o) ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, Buparlisib, Pictrellisib, PF-4691502, BYL-719, Doctorisib, XL-147, Compounds that target, reduce or inhibit the kinase activity of PI3 kinase (PI3K), including but not limited to XL-765 and idelalisib; and; and q) signaling effects of the hedgehog protein (Hh) or smooth receptor (SMO) pathways, including but not limited to cyclopamine, vismodegib, itraconazole, erysmodegib, and IPI-926 (saridegib). Includes, but is not limited to, compounds that target, reduce or inhibit .

As used herein, the term “PI3K inhibitor” includes 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. , but is not limited to this. Examples of PI3K inhibitors useful in the present invention include ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, Bupalisib, Pictrellisib, PF-4691502, BYL-719, Doc. Includes, but is not limited to, Toryship, XL-147, XL-765, and Idelariship.

As used herein, the term "Bcl-2 inhibitor" includes ABT-199, ABT-731, ABT-199, ABT-731, ABT-737, apogosypol, Ascenta's pan-Bcl-2 inhibitor, curcumin ( and analogues thereof), dual Bcl-2/Bcl-xL inhibitor (Infinity Pharmaceuticals/Novartis Pharmaceuticals), Genasense (G3139), HA14-1 (and analogues thereof; see WO2008118802), navitoclax (and analogues thereof, US7390799) Reference), NH-1 (Shenayng Pharmaceutical University), Obatoclax (and its analogues, see WO2004106328), S-001 (Gloria Pharmaceuticals), TW series compounds (Univ. of Michigan), and venetoclax , but is not limited to these. 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 with inhibitory activity against Bruton's tyrosine kinase (BTK), including but not limited to AVL-292 and ibrutinib. It doesn't work.

As used herein, the term “SYK inhibitor” refers to spleen tyrosine kinase (SYK) inhibitors, including but not limited to PRT-062070, R-343, R-333, Excellair, PRT-062607, and fostamatinib. Including, but not limited to, compounds having inhibitory activity against.

Additional examples of BTK inhibitory compounds, and conditions treatable by such compounds in conjunction with the compounds of the present invention, can be found in WO2008039218 and WO2011090760, the entire patent documents being incorporated herein by reference.

Additional examples of SYK inhibitory compounds, and conditions treatable by such compounds in conjunction with the compounds of the present invention, can be found in WO2003063794, WO2005007623, and WO2006078846, the entire patent documents being incorporated herein by reference.

Additional examples of PI3K inhibitory compounds and conditions treatable by such compounds in combination with the compounds of the present invention include WO2004019973, WO2004089925, WO2007016176, US8138347, WO2002088112, WO2007084786, WO2007129161, WO2006122 806, WO 2005113554, and WO2007044729, above. The entire patent document is incorporated herein by reference.

Additional examples of JAK inhibitory compounds, and conditions treatable by such compounds in conjunction with the compounds of the invention, can be found in WO2009114512, WO2008109943, WO2007053452, WO2000142246, and WO2007070514, the entire patent document being incorporated herein by reference. .

Additional anti-angiogenic compounds include compounds that have alternative mechanisms for their activity, e.g., those not associated with protein or lipid kinase inhibition, such as thalidomide (THALOMID™) and TNP-470. .

Examples of proteasome inhibitors useful for use with the compounds of the invention include bortezomib, disulfiram, epigallocatechin-3-gallate (EGCG), salinosporamide A, carfilzomib, ONX-0912. , CEP-18770, and 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 derivatives thereof.

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 includes Cox-2 inhibitors, 5-alkyl substituted 2-arylaminophenylacetic acids and derivatives such as CELEBREX™, rofecoxib (VIOXX™), etoricoxib, valdecoxib, or 5-alkyl-2-arylaminophenylacetic acid, such as 5-methyl-2-(2'-chloro-6'-fluoroanilino)phenyl acetic acid, including lumiracoxib However, it is not limited to these.

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. No. Etridonic acid is marketed under the trade name DIDRONEL™. Clodronic acid is marketed under the trade name BONEFOS™. Tiludronic acid is marketed under the trade name Skelid™. Pamidronic acid is marketed under the trade name AREDIA™. Alendronic acid is marketed under the brand name FOSAMAX™. Ibandronic acid is marketed under the trade name BONDRANAT™. Risedronic acid is marketed under the brand name ACTONEL™. Zoledronic acid is marketed under the brand name ZOMETA™. 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 ABT578. will be.

As used herein, the term “heparanase inhibitor” refers to a compound that targets, reduces or inhibits heparin sulfate degradation. The term includes, but is not limited to, PI-88. As used herein, the term “biological response modifier” refers to 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 a compound that targets, reduces or inhibits the oncogenic activity of Ras; Refers to a “farnesyl transferase inhibitor”, for example, 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 those that inhibit the telomerase receptor, particularly telomestatin.

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

As used herein, the term “telomerase inhibitor” refers to a compound that targets, reduces or inhibits the activity of the proteasome. Compounds that target, reduce or inhibit the activity of the proteasome include, but are not limited to, bortezomib (VELCADE™) and MLN 341.

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

As used herein, the term “compounds used in the treatment of hematologic malignancies” includes FMS-like tyrosine kinase inhibitors, which are compounds that target and reduce or inhibit the activity of the FMS-like tyrosine kinase receptor (Flt-3R); interferon, 1-β-D-arabinofuransylcytosine (ara-c) and bisulfan; and ALK inhibitors, which are compounds that target, reduce or inhibit anaplastic lymphoma kinase.

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, such as PKC412, midostaurin, star Urosporine derivatives, SU11248 and MLN518.

As used herein, the term “HSP90 inhibitor” refers to a compound that targets, reduces or inhibits the intrinsic ATPase activity of HSP90; Includes, but is not limited to, compounds that degrade, target, reduce or inhibit HSP90 client proteins through the ubiquitin proteosome pathway. Compounds that target, reduce or inhibit the intrinsic ATPase activity of HSP90 include compounds, proteins or antibodies that specifically inhibit the ATPase activity of HSP90, such as 17-allylamino, 17-demethoxygeldanamycin (17AAG), Geldana Mycin derivatives; Other geldanamycin-related compounds; It is a radicicol and HDAC inhibitor.

As used herein, the term “antiproliferative antibody” refers to TRAstuzumab (HERCEPTIN™), Trastuzumab-DM1, Erbitux, Bevacizumab (AVASTIN™), Rituximab ( ^RITUXAN® ), PRO64553 (anti-CD40) and 2C4 antibodies. Antibody refers to intact monoclonal antibodies, polyclonal antibodies, multispecific antibodies formed from at least two intact antibodies, and antibody fragments that 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, especially those used in the treatment of AML. In particular, the compounds of the invention may be used in combination with, for example, farnesyl transferase inhibitors and/or other drugs useful in the treatment of AML, such as daunorubicin, adriamycin, Ara-C, VP-16, teniposide, mitoxantrone, Can be administered with idarubicin, carboplatinum, and PKC412.

Other anti-leukemia compounds include, for example, the pyrimidine analog, Ara-C, which is a 2-alpha-hydroxy ribose (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 suberoylanilide hydroxamic acid (SAHA), inhibit the activity of enzymes known as histone deacetylators. Specific HDAC inhibitors include MS275, SAHA, FK228 (formerly FR901228), Trichostatin A, and N-hydroxy-3-[4-[[[2-(2-methyl-1H-indol-3-yl)-ethyl] -amino]methyl]phenyl]-2E-2-propenamide, or a pharmaceutically acceptable salt thereof and N-hydroxy-3-[4-[(2-hydroxyethyl){2-(1H-indole -3-yl)ethyl]-amino]methyl]phenyl]-2E-2-propenamide, or pharmaceutically acceptable salts thereof, especially the lactate salt, including but not limited to the compounds disclosed in US 6,552,065. Includes. As used herein, somatostatin receptor antagonist refers to compounds that target, treat or inhibit the somatostatin receptor, such as octreotide and SOM230. Tumor cell damaging approaches refer to approaches such as ionizing radiation. The term "ionizing radiation", as mentioned above and below, means ionizing radiation that occurs as electromagnetic waves (such as X-rays and gamma rays) or particles (such as alpha and beta particles). Ionizing radiation is provided in, but is not limited to, radiation therapy and is known in the art. Hellman, Principles of Radiation Therapy, Cancer, in Principles and Practice of Oncology, Devita et al., Eds., 4 ^th Edition, Vol. 1, pp. 248-275 (1993).

EDG binders and ribonucleotide reductase inhibitors are also included. As used herein, the term “EDG binder” refers to a class of immunosuppressive agents that modulate lymphocyte recycling, such as FTY720. The term "ribonucleotide reductase inhibitor" refers to fludarabine and/or cytosine arabinoside (ara-C), 6-thioguanine, 5-fluorouracil, cladribine, 6-mercaptopurine (particularly for ALL). together with ara-C) and/or 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; or anti-VEGF antibodies such as rhuMAb and RHUFab or anti-VEGF receptor antibodies, VEGF aptamers such as Macugon; Also included are such compounds, proteins or monoclonal antibodies of VEGF such as FLT-4 inhibitors, FLT-3 inhibitors, VEGFR-2 IgGI antibodies, Anziozyme (RPI 4610) and bevacizumab (AVASTIN™).

As used herein, photodynamic therapy refers to therapy that uses certain chemicals known as photosensitive compounds to treat or prevent cancer. Examples of photodynamic therapy include treatment with compounds such as VISUDYNE™ and porphymer sodium.

As used herein, anti-angiogenic steroids include, for example, anecortabe, triamcinolone, hydrocortisone, 11-α-epihydrocortisol, cortexolone, 17α-hydroxyprogesterone, corticosterone, desoxycortisone. Refers to compounds that block or inhibit angiogenesis, such as costerone, testosterone, estrone, and dexamethasone.

Corticosteroids containing implants refer to compounds such as fluocinolone and dexamethasone.

Other chemotherapy compounds include plant alkaloids, hormonal compounds, and antagonists; Biological response modifiers, preferably lymphokines or interferons; Antisense oligonucleotides or oligonucleotide derivatives; shRNA or siRNA; or miscellaneous compounds, or compounds with different or unknown mechanisms of action.

The structures of the active compounds identified by code number, generic name or trade name may be taken from the actual version of the standard compendium "The Merck Index" or from databases, such as international patents (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-oncology 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 together with a compound of the invention has a synergistic effect in the treatment of cancer.

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

In some embodiments, the immuno-oncological agent is (i) an agonist of stimulatory (including co-stimulatory) receptors or (ii) an antagonist of inhibitory (including co-inhibitory) signals on T cells, where both (i) and (ii) Amplifies 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 co-stimulatory or co-inhibitory receptors is B7-1, B7-2, B7-H1 (PD-L1), B7-DC (PD-L2), and B7-H2 (ICOS). -L), B7-H3, B7-H4, B7-H5 (VISTA), and B7-H6. Another family of membrane-bound ligands that bind to co-stimulatory or co-inhibitory receptors are CD40 and CD40L, OX-40, OX-40L, CD70, CD27L, CD30, CD30L, 4-1BBL, CD137(4-1BB), and 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, (cognate) A member of the TNF family of molecules that binds to members of the TNF receptor family.

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

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

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

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; WO14/ 036357) Agents that inhibit or deplete macrophages or monocytes, including but not limited to CSF-1R antagonists, such as CSF-1R antagonist antibodies.

In some embodiments, the immuno-oncology agent is an agonistic agent that ligates positive costimulatory receptors, a blocker that attenuates signaling through inhibitory receptors, an antagonist, and one or more agents that systemically increase the frequency of anti-tumor T cells. , conquering distinct immunosuppressive pathways within the tumor microenvironment (e.g., blocking inhibitory receptor binding (e.g., PD-L1/PD-1 interaction), depleting or suppressing Tregs ( (e.g., using anti-CD25 monoclonal antibodies (e.g., daclizumab) or by anti-CD25 bead depletion in vitro), inhibiting metabolic enzymes such as IDO, or reversing/reversing T cell energy or exhaustion. agents that prevent) and 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 antagonistic CTLA-4 antibody. In some embodiments, the antagonistic CTLA-4 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 death-1 (PD-1) receptor and inhibits PD-1 activity. In some embodiments, the PD-1 antagonist is an antagonistic PD-1 antibody. In some embodiments, the antagonistic PD-1 antibody is OPDIVO (nivolumab), KEYTRUDA (pembrolizumab), or MEDI-0680 (AMP-514; WO2012/145493). In some embodiments, the immuno-oncology agent is pidilizumab (CT-011). In some embodiments, the immuno-oncology agent is a recombinant protein consisting of the extracellular domain of PD-L2 (B7-DC) fused to the Fc portion of IgG1, called AMP-224.

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

In some embodiments, the immuno-oncology agent is a LAG-3 antagonist. In some embodiments, the LAG-3 antagonist is an antagonistic LAG-3 antibody. In some embodiments, the 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 a functional CD137 antibody. In some embodiments, the CD137 antibody is urelumab or PF-05082566 (WO12/32433).

In some embodiments, the immuno-oncology agent is a GITR agonist. In some embodiments, the GITR agonist is a functional CITR 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-oncological 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 (Genentech/ROChe); PF-06840003 (Pfizer); BMS:F001287 (Bristol-Myers Squibb); Phy906/KD108 (PhytOCeutica); An enzyme that breaks down kynurenine (Kynase, Ikena Oncology, formerly known as Kyn Therapeutics); and NLG-919 (WO09/73620, WO009/1156652, WO11/56652, WO12/142237).

In some embodiments, the immuno-oncology agent is an OX40 agonist. In some embodiments, the OX40 agonist is a functional 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 antagonistic 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 a functional CD40 antibody. In some embodiments, the immuno-oncology agent is a CD40 antagonist. In some embodiments, the CD40 antagonist is an antagonistic CD40 antibody. In some embodiments, the CD40 antibody is rucatumumab or dacetuzumab.

In some embodiments, the immuno-oncology agent is a CD27 agonist. In some embodiments, the CD27 agonist is a functional CD27 antibody. In some embodiments, the CD27 antibody is barilumab.

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

In some embodiments, the immuno-oncology agent is abagovomab, adecatumumab, aputuzumab, alemtuzumab, anatumomab mapenatox, apolizumab, atezolimab, avelumab, blinatumomab, BMS-936559 , catumaxomab, durvalumab, epacadostat, epratuzumab, indoximod, inotuzumab ozogamicin, intelumumab, ipilimumab, isatuximab, lambrolizumab, MED14736, MPDL3280A, Nivolumab, obinutuzumab, ocaratuzumab, ofatumumab, olatumab, pembrolizumab, pidilizumab, rituximab, ticilimumab, samalizumab, or tremelimumab.

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

In some embodiments, the immunomodulatory therapeutic agent specifically induces apoptosis of tumor cells. Approved immunomodulatory treatments that can be used in the present invention include pomalidomide (POMALYST®, Celgene); lenalidomide (REVLIMID®, Celgene); Includes ingenol mebutate (PICATO®, LEO Pharma).

In some embodiments, the immuno-oncology agent is a cancer vaccine. In some embodiments, the cancer vaccine is sipuleucel-T (PROVENGE®, Dendreon/Valeant Pharmaceuticals), which has been approved for the treatment of asymptomatic, or minimally symptomatic, metastatic castration-resistant (hormone-refractory) prostate cancer; and talimogene laherparepvec (IMLYGIC®, BioVex/Amgen, formerly T-VEC), a genetically modified oncolytic viral therapy approved for the treatment of unresectable cutaneous, subcutaneous, and lymph node lesions in melanoma. is selected from In some embodiments, the immuno-oncology agent is an oncolytic viral therapy, such as Peck, a thymidine kinase-(TK-)-deficient vaccinia virus engineered to express GM-CSF for hepatocellular carcinoma (NCT02562755) and melanoma (NCT00429312). pexastimogene devacirepvec (PexaVec/JX-594, SillaJen/formerly Jennerex Biotherapeutics); Colorectal cancer (NCT01622543), prostate cancer (NCT01619813); Head and neck squamous cell carcinoma (NCT01166542); Pancreatic adenocarcinoma (NCT00998322); and REOLYSIN®, Oncolytics Biotech, a variant of a respiratory intestinal reovirus that does not replicate in non-RAS-activated cells in a number of cancers, including non-small cell lung cancer (NSCLC) (NCT 00861627); In metastatic or advanced epithelial tumors, such as ovarian cancer (NCT02028117), colorectal cancer, bladder cancer, head and neck squamous cell carcinoma, and salivary gland cancer (NCT02636036), adenoviruses engineered to express antibody fragments specific for full-length CD80 and T cell receptor CD3 proteins. In Enadenotushirev (NG-348, PsiOxus, formerly known as ColoAd1); Melanoma (NCT03003676); and ONCOS-102 (Targovax/formerly Oncos), an adenovirus engineered to express GM-CSF in peritoneal disease, colorectal or ovarian cancer (NCT02963831); Peritoneal carcinomatosis (NCT01443260); Beta-galactosidase (beta-gal)/beta-glucoronidase or beta-gal/human sodium iodide symporter (hNIS), respectively, which have been studied in fallopian tube and ovarian cancer (NCT 02759588) GL-ONC1 (GLV-1h68/GLV-1h153, Genelux GmbH), a vaccinia virus engineered to express . or CG0070 (Cold Genesys), an adenovirus engineered to express GM-CSF in bladder cancer (NCT02365818).

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

In some embodiments, the immuno-oncology agent is a T-cell engineered to express a chimeric antigen receptor, or CAR. T cells engineered to express these chimeric antigen receptors are referred to as CAR-T cells.

A single antibody derived from a monoclonal antibody specific for a cell surface antigen fused to the endodomain, the functional end of the T cell receptor (TCR), such as the CD3-zeta signaling domain from the TCR, capable of generating an activation signal in T lymphocytes. A CAR consisting of a binding domain that can be derived from a natural ligand, a chain variable fragment (scFv), has been constructed. Upon antigen binding, these CARs connect to endogenous signaling pathways in the effector cell and generate activation signals similar to those initiated by the TCR complex.

For example, in some embodiments CAR-T cells are extracellular with an antigen binding domain (e.g., a domain that binds CD19) fused to the intracellular signaling domain of the T cell antigen receptor complex zeta chain (e.g., CD3 zeta). One of those described in US Pat. No. 8,906,682 (June et al.; incorporated herein by reference in its entirety) discloses CAR-T cells engineered to contain the domain. When expressed in T cells, CARs can redirect antigen recognition based on antigen binding specificity. In the case of CD19, the antigen is expressed on malignant B cells. More than 200 clinical trials are currently underway using CAR-T in a variety 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 receptorγ (RORγt). RORγt is a transcription factor that 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. am. In some embodiments, the activator of RORγt is LYC-55716 (Lycera), which is currently being evaluated in a clinical trial for the treatment of solid tumors (NCT02929862).

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

Other immuno-oncology agents that may be used in the present invention include urelumab (BMS-663513, Bristol-Myers Squibb), an anti-CD137 monoclonal antibody; barilumab (CDX-1127, Celldex Therapeutics), an anti-CD27 monoclonal antibody; BMS-986178, an anti-OX40 monoclonal antibody (Bristol-Myers Squibb); ririlumab (IPH2102/BMS-986015, Innate Pharma, Bristol-Myers Squibb), an anti-KIR monoclonal antibody; Monalizumab (IPH2201, Innate Pharma, AstraZeneca), an anti-NKG2A monoclonal antibody; Andecaliximab (GS-5745, Gilead Sciences), an anti-MMP9 antibody; Includes MK-4166 (Merck & Co.), an anti-GITR monoclonal antibody.

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

In some embodiments, the immunostimulatory therapeutic agent is recombinant human interleukin 15 (rhIL-15). rhIL-15 has been tested clinically 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, the IL-15 based immunotherapeutic agent comprises the soluble IL-15 binding protein IL-15, tested in a phase 1 clinical trial for melanoma, renal cell carcinoma, non-small cell lung cancer, and head and neck squamous cell carcinoma (NCT02452268). It is heterodimeric IL-15 (hetIL-15, Novartis/Admune), a fusion complex consisting of a synthetic form of endogenous IL-15 complexed to the receptor alpha chain (IL15:sIL-15RA). 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 as described in Jerry L. Adams et al., “Big opportunities for small molecules in immuno-oncology,” Cancer Therapy 2015 , Vol. 14, pages 603-622, the entire contents of which are incorporated herein by reference. In some embodiments, the immuno-oncology agent is selected from the examples set forth in Table 1 of Jerry L. Adams et al. In some embodiments, the immuno-oncology agent is a small molecule that targets an immuno-oncology agent target selected from those listed in Table 2 of Jerry L. Adams et al. In some embodiments, the immuno-oncology agent is a small molecule agent selected from those listed in Table 2 of Jerry L. Adams et al.

In some embodiments, the immuno-oncology agent is an immuno-oncology agent as described in Peter L. Toogood, “Small molecule immuno-oncology therapeutic agents,” Bioorganic & Medicinal Chemistry Letters 2018 , Vol. 28, pages 319-329, the entire contents of which are incorporated herein by reference. In some embodiments, the immuno-oncological agent is an agent that targets a pathway as described in Peter L. Toogood.

In some embodiments, the immuno-oncology agent is selected from those described in Sandra L. Ross et al., "Bispecific T cell engager (BiTE®) antibody constructs can mediate bystander tumor cell killing", PLoS ONE 12(8): e0183390, The entire contents of the above document are incorporated herein by reference. 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 to release cytokines that induce upregulation of intercellular adhesion molecule 1 (ICAM-1) and FAS on bystander cells. . In some embodiments, the bispecific T cell engager (BiTE®) antibody construct activates T cells leading to bystander cell lysis. In some embodiments, the bystander cells are in a solid tumor. In some embodiments, the bystander cells that are lysed are adjacent to BiTE® activated T cells. In some embodiments, bystander cells comprise tumor associated antigen (TAA) negative cancer cells. In some embodiments, bystander cells comprise EGFR-negative cancer cells. In some embodiments, the immuno-oncology agent is an antibody that blocks the PD-L1/PD1 axis and/or CTLA4. In some embodiments, the immuno-oncological agent is tumor-infiltrating T cells expanded ex vivo. In some embodiments, the immuno-oncology agent is a bispecific antibody construct or chimeric antigen receptor (CAR) that directly engages 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 as 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 antitumor immune subversion is known as “T cell exhaustion”, which occurs due to chronic exposure to antigens that result in upregulation of inhibitory receptors. These inhibitory receptors act 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 usually referred to as checkpoint regulators. They are extracellular information that dictates whether cell cycle progression and other intracellular signaling processes should proceed. It acts as a molecule “gatekeeper” that allows .

In some embodiments, the inhibitor of an immune checkpoint is an antibody against PD-1. PD-1 binds to the programmed cell death 1 receptor (PD-1), preventing the receptor from binding to the inhibitory ligand PDL-1, overriding the tumor's ability to suppress host anti-tumor immune responses.

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, CHK 1 , CHK2, A2aR, B-7 family ligands, or combinations 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, CHK 1 , CHK2, A2aR, B-7 family ligands, or combinations thereof. In one aspect, the checkpoint inhibitor is an immunostimulatory agent, T cell growth factor, interleukin, antibody, vaccine, or combination thereof. In a further aspect, the interleukin is IL-7 or IL-15. In certain aspects, 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 inhibitory pathways of the immune system in a statistically significant manner. Such inhibitors may include small molecule inhibitors, or may include antibodies or antigen-binding fragments thereof that bind to and block or inhibit an immune checkpoint receptor, or antibodies that bind to and block or inhibit an immune checkpoint receptor ligand. . Exemplary checkpoint molecules that can be targeted for blocking 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 referred to as BY55), CGEN-15049, CHK 1 and CHK2 kinases, A2aR, and various B-7 Includes, but is not limited to, family ligands. B7 family ligands include, but are limited to, B7-1, B7-2, B7-DC, B7-H1, B7-H2, B7-H3, B7-H4, B7-H5, B7-H6, and B7-H7 It doesn't work. Checkpoint inhibitors are antibodies that bind to and block or inhibit the activity of one or more of CTLA-4, PDL1, PDL2, PD1, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD 160, and CGEN-15049. , or an antigen-binding fragment thereof, another binding protein, biological therapeutic, or small molecule. Exemplary immune checkpoint inhibitors include tremelimumab (CTLA-4 blocking antibody), anti-OX40, PD-L1 monoclonal antibody (anti-B7-H1; MEDI4736), MK-3475 (PD-1 blocker), nivolumab ( anti-PD1 antibody), CT-011 (anti-PD1 antibody), BY55 monoclonal antibody, AMP224 (anti-PDL1 antibody), BMS-936559 (anti-PDL1 antibody), MPLDL3280A (anti-PDL1 antibody), MSB0010718C (anti-PDL1 antibody) PDL1 antibody), and ipilimumab (anti-CTLA-4 checkpoint inhibitor). Checkpoint protein ligands include, but are not limited to, PD-L1, PD-L2, B7-H3, B7-H4, CD28, CD86, and TIM-3.

In certain embodiments, the immune checkpoint inhibitor is selected from PD-1 antagonists, PD-L1 antagonists, and CTLA-4 antagonists. In some embodiments, the checkpoint inhibitor is selected from the group consisting of nivolumab (OPDIVO®), ipilimumab (YERVOY®), and pembrolizumab (KEYTRUDA®). In some embodiments, the checkpoint inhibitor is nivolumab (anti-PD-1 antibody, OPDIVO®, Bristol-Myers Squibb); Pembrolizumab (anti-PD-1 antibody, KEYTRUDA®, Merck); Ipilimumab (anti-CTLA-4 antibody, YERVOY®, Bristol-Myers Squibb); durvalumab (anti-PD-L1 antibody, IMFINZI®, AstraZeneca); and atezolizumab (anti-PD-L1 antibody, TECENTRIQ®, Genentech).

In some embodiments, the checkpoint inhibitor is lambrolizumab (MK-3475), nivolumab (BMS-936558), pidilizumab (CT-011), AMP-224, MDX-1105, MEDI4736, MPDL3280A, BMS-936559. , ipilimumab, ririlumab, IPH2101, pembrolizumab (KEYTRUDA®), and tremelimumab.

In some embodiments, the immune checkpoint inhibitor is selected from the group consisting of basal cell carcinoma (NCT03132636) NSCLC (NCT03088540); Cutaneous squamous cell carcinoma (NCT02760498); Lymphoma (NCT02651662); and REGN2810 (Regeneron), an anti-PD-1 antibody tested in patients with melanoma (NCT03002376); In clinical trials for diffuse large B-cell lymphoma and multiple myeloma, pidilizumab (CureTech), also known as CT-011, an antibody that binds to PD-1; Avelumab (BAVENCIO®, Pfizer), a fully human IgG1 anti-PD-L1 antibody, in clinical trials for non-small cell lung cancer, Merkel cell carcinoma, mesothelioma, solid tumors, kidney cancer, ovarian cancer, bladder cancer, head and neck cancer, and gastric cancer. /Merck KGaA), also known as MSB0010718C); or PDR001 (Novartis), an inhibitory antibody that binds to PD-1, in clinical trials for non-small cell lung cancer, melanoma, triple negative breast cancer, and advanced or metastatic solid tumors. Tremelimumab (CP-675,206; Astrazeneca) is used to treat mesothelioma, colorectal cancer, kidney cancer, breast cancer, lung cancer and non-small cell lung cancer, pancreatic adenocarcinoma, pancreatic cancer, germ cell cancer, squamous cell carcinoma of the head and neck, hepatocellular carcinoma, prostate cancer, and uterus. Clinical trials for multiple indications, including endometrial cancer, liver metastases, liver cancer, large B-cell lymphoma, ovarian cancer, cervical cancer, metastatic anaplastic thyroid cancer, urothelial cancer, fallopian tube cancer, multiple myeloma, bladder cancer, soft tissue sarcoma, and melanoma. It is a fully human monoclonal antibody against CTLA-4 studied in trials. AGEN-1884 (Agenus) is an anti-CTLA4 antibody being investigated in a phase 1 clinical trial for 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 investigated in solid tumors (NCT02817633). LY3321367 (Eli Lilly) is an anti-TIM-3 antibody being investigated in solid tumors (NCT03099109). MBG453 (Novartis) is an anti-TIM-3 antibody being investigated 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 immune receptor on certain T cells and NK cells. TIGIT inhibitors that can be used in the present invention include BMS-986207 (Bristol-Myers Squibb), an 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. The anti-LAG-3 antibody, BMS-986016 (Bristol-Myers Squibb), is being studied in glioblastoma and gliosarcoma (NCT02658981). REGN3767 (Regeneron) is also an anti-LAG-3 antibody, which is being studied in malignancies (NCT03005782). IMP321 (Immutep S.A.) was used in melanoma (NCT02676869); Adenocarcinoma (NCT02614833); and the LAG-3-Ig fusion protein being studied in 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 agonistic anti-OX40 antibody in metastatic renal cancer (NCT03092856) and advanced cancers and neoplasms (NCT02554812; NCT05082566); GSK3174998 (Merck), an agonistic anti-OX40 antibody, in a phase 1 cancer trial (NCT02528357); MEDI0562 (Medimmune/AstraZeneca), an agonistic anti-OX40 antibody in advanced solid tumors (NCT02318394 and NCT02705482); MEDI6469, an agonistic anti-OX40 antibody (Medimmune/AstraZeneca) in patients with colorectal cancer (NCT02559024), breast cancer (NCT01862900), head and neck cancer (NCT02274155), and metastatic prostate cancer (NCT01303705); and BMS-986178 (Bristol-Myers Squibb), an agonistic anti-OX40 antibody in advanced cancer (NCT02737475).

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

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

Checkpoint inhibitors that can be used in the present invention include glucocorticoid-induced tumor necrosis factor receptor (GITR) agonists. GITR agonists being investigated 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 agonistic anti-GITR antibody in solid tumors and lymphomas (NCT 02740270); INCAGN01876 (Incyte/Agenus), an agonistic anti-GITR antibody in advanced cancer (NCT02697591 and NCT03126110); MK-4166 (Merck), an agonistic anti-GITR antibody in solid tumors (NCT02132754); and MEDI1873 (Medimmune/AstraZeneca), a functional hexameric GITR-ligand molecule with a human IgG1 Fc domain in advanced solid tumors (NCT02583165).

Checkpoint inhibitors that may be used in the present invention include inducible T cell costimulator (ICOS, also known as CD278) agonists. ICOS agents being investigated in clinical trials include MEDI-570 (Medimmune), an agonistic anti-ICOS antibody in lymphoma (NCT02520791); GSK3359609 (Merck), an agonistic anti-ICOS antibody, in phase 1 (NCT02723955); and JTX-2011 (Jounce Therapeutics), an agonistic anti-ICOS antibody in phase 1 (NCT02904226).

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

Checkpoint inhibitors that can be used in the present invention include CD47 inhibitors of the interaction between CD47 and signal regulatory protein alpha (SIRPa). CD47/SIRPa inhibitors being studied in clinical trials include ALX-148 (Alexo Therapeutics), an antagonistic variant of (SIRPa) that binds CD47 and prevents CD47/SIRPa-mediated signaling in phase 1 (NCT03013218); It is a soluble recombinant fusion protein created by linking the N-terminal CD47 binding domain of SIRPa and the Fc domain of human IgG1, and serves to bind to human CD47 and prevent it from transmitting its “do not eat” signal to macrophages. TTI-621 (SIRPa-Fc, Trillium Therapeutics), in clinical trials in phase 1 (NCT02890368 and NCT02663518); CC-90002 (Celgene), an anti-CD47 antibody in leukemia (NCT02641002); and Hu5F9-G4 (Forty Seven, Inc.) in colorectal neoplasms 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 investigated in clinical trials include MEDI9447 (Medimmune), an anti-CD73 antibody in solid tumors (NCT02503774); and BMS-986179 (Bristol-Myers Squibb), an anti-CD73 antibody in solid tumors (NCT02754141).

Checkpoint inhibitors that may be used in the present invention include agonists of the stimulator of interferon gene protein (STING, also known as transmembrane protein 173, or TMEM173). Agonists of STING being studied in clinical trials include MK-1454 (Merck), a synthetic cyclic dinucleotide functional in lymphoma (NCT03010176); and ADU-S100 (MIW815, Aduro Biotech/Novartis), a functional synthetic cyclic dinucleotide in phase 1 (NCT02675439 and NCT03172936).

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

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

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

example

The following examples are intended to illustrate the invention and should not be construed as limiting its scope. Unless otherwise stated, one or more tautomeric forms of the compounds of the examples described below can be prepared and/or isolated in situ. All tautomeric forms of the compounds of the examples described below should be considered disclosed. Temperatures are given 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 structures of the final products, intermediates and starting materials are confirmed by standard analytical methods, such as microanalysis and spectroscopic characterization, such as MS, IR, NMR. The abbreviations used are those that are conventional in the art.

All starting materials, building blocks, reagents, acids, bases, dehydrating agents, solvents, and catalysts used to synthesize the compounds of the present invention are commercially available or can be produced by organic synthesis methods known to those skilled in the art. Additionally, the compounds of the present invention can be produced by organic synthesis methods known to those skilled in the art, as shown in the examples below.

Example 1: Synthesis of Exemplary Compounds

Certain exemplary compounds are prepared according to the scheme below.

I-1

Step 1 : [3-(trifluoromethyl)-1-bicyclo[1.1.1]fentanyl]methanol

3-(trifluoromethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (200 mg, 1.11 mmol) in a solution of LiAlH ₄ (84.28 mg, 2.22 mmol, 2 eq) in THF (5 mL) at 0°C. , 1 eq) was added and stirred at 25°C for 2 hours. TLC (PE/EtOAc = 1/1, R _f = 0.60) showed that the starting material was completely consumed and one new spot was formed. 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]fentanyl]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. ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 3.67 (s, 2H), 1.92 (s, 6H).

Step 2 : [3-(trifluoromethyl)-1-bicyclo[1.1.1]fentanyl]methyl methanesulfonate

[3-(trifluoromethyl)-1-bicyclo[1.1.1]fentanyl]methanol (180 mg, 975.08 μmol, 90% purity, 1 eq) and Et ₃ N ( MsCl (170 mg, 1.48 mmol, 114.86 μL, 1.52 eq) was added dropwise to a solution of 197.33 mg, 1.95 mmol, 271.44 μL, 2 eq), and the mixture was stirred at 25°C for 2 hours. TLC (PE/EtOAc = 1/1, R _f = 0.67) showed that the starting material was completely consumed and one new spot was formed. The mixture was quenched with saturated aqueous NaHCO ₃ (10 mL) and extracted using DCM (15 mLx 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]fentanyl]methyl methanesulfonate (230 mg, 894.65 μmol, 91.8 μmol). % yield, 95.0% purity) was obtained as a colorless gum, which was used in the next step without further purification. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 4.25 (s, 2H), 3.02 (s, 3H), 2.02 (s, 6H).

Step 3 : N -[(1 S ,5 R )-3-[[3-(trifluoromethyl)-1-bicyclo[1.1.1]fentanyl]methyl]-3-azabicyclo[3.1.0 ]hexan-6-yl]prop-2-enamide

N -[(1 S ,5 R )-3-azabicyclo[3.1.0]hexan-6-yl]prop-2-enamide (120 mg, 360.61 μmol, 80% purity) in ACN (3 mL) , 1 eq, TFA) and [3-(trifluoromethyl)-1-bicyclo[1.1.1]fentanyl]methyl methanesulfonate (92.71 mg, 360.61 μmol, 95% purity, 1 eq). 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 -78%, 10 minutes) and then lyophilized to form N -[(1 S ,5 R )-3-[[3-(trifluoromethyl)-1-bicyclo[1.1.1]fentanyl]methyl. ]-3-azabicyclo[3.1.0]hexan-6-yl]prop-2-enamide (18.28 mg, 60.87 μmol, 16.9% yield, 100.0% purity) was obtained as a white solid. ¹ H 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 = 10.2 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] ⁺ .

I-8

Step 1: [4-(trifluoromethyl)-1-bicyclo[2.2.2]octanyl]methanol

LiAlH ₄ (102.49 mg, 2.70 mmol) in a solution of 4-(trifluoromethyl)bicyclo[2.2.2]octane-1-carboxylic acid (300 mg, 1.35 mmol, 1 eq) in THF (5 mL) at 25°C. , 2 eq) was added. The mixture was stirred at 25°C for 1 hour. TLC (PE/EtOAc = 3/1, R _f = 0.59) showed that the starting material was completely consumed and one new spot was formed. 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 mmol, 94.6% yield, 95.0% purity) was obtained as a colorless gum, 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).

Step 2 : 4-(trifluoromethyl)bicyclo[2.2.2]octane-1-carbaldehyde

PCC (98.35 mg) was added to a solution of [4-(trifluoromethyl)-1-bicyclo[2.2.2]octanyl]methanol (50 mg, 228.12 μmol, 95% purity, 1 eq) in DCM (3 mL). , 456.25 μmol, 2 eq) was added. The mixture was stirred at 25°C for 1 hour. TLC (PE/EtOAc = 3/1, R _f = 0.75) showed that the starting material was completely consumed and one new spot was formed. The solvent was removed to give 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 give 4-(trifluorocarbon Methyl)bicyclo[2.2.2]octane-1-carbaldehyde (50 mg, 223.08 μmol, 97.8% yield, 92.0% purity) 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).

Step 3: N -[(One R ,5 S )-3-[[4-(trifluoromethyl)-1-bicyclo[2.2.2]octanyl]methyl]-3-azabicyclo[3.1.0]hexan-6-yl]prop-2 -Enamide

N -[(1 S ,5 R )-3-azabicyclo[3.1.0]hexan-6-yl]prop-2-enamide (120 mg, 360.61 μmol, 80% purity) in MeOH (10 mL) , 1 eq, TFA) and 4-(trifluoromethyl)bicyclo[2.2.2]octane-1-carbaldehyde (47.43 mg, 211.62 mg) in a solution of TEA (36.49 mg, 360.61 μmol, 50.19 μL, 1 eq). μmol, 92% purity) 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 * 25 mm * 5 um; mobile phase: [water (0.05% NH ₃ H ₂ O + 10 mM NH ₄ HCO ₃ )- ACN]; B%: 51%-81%, 10 minutes) and then lyophilized to form N -[(1 S ,5 R )-3-[[4-(trifluoromethyl)-1-bi. Cyclo[2.2.2]octanyl]methyl]-3-azabicyclo[3.1.0]hexan-6-yl]prop-2-enamide (24.64 mg, 71.96 μmol, 20.0% yield, 100.0% purity) was obtained as a white solid. ^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 = 1.7 Hz, 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] ⁺ .

I-9

Step 1 : tert -Butyl (2-bromo-4-nitrophenyl)(4-(trifluoromethyl)benzyl)carbamate

To 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 (from PE/EtOAc = 1/0 to 10/1, TLC: PE/EtOAc = 10/1, R _f = 0.51) to give tert -Butyl N -(2-bromo-4-nitro-phenyl)- N -[[4-(trifluoromethyl)phenyl]methyl]carbamate (1.15 g, 2.40 mmol, 89.9% yield, 98.7% purity) was obtained as a white solid. ^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] ⁺ .

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 in EtOH (5 mL) and H ₂ O (5 mL) (1.11 g, 2.31 mmol, 98.7%, 1 eq) was 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 using EtOAc (30 mL x 3). The combined organic layers were washed with brine (10 mL), dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure 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 used in the next step without further purification. ^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 3 : tert -Butyl (4-acrylamido-2-bromophenyl)(4-(trifluoromethyl)benzyl)carbamate

tert -Butyl N- (4-amino-2-bromo-phenyl) -N -[[4-(trifluoromethyl)phenyl]methyl]carbamate (250 mg, 539.00 μmol, 96%) in DCM (10 mL) 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) were added to the solution (% purity, 1 eq). The mixture was stirred at 25°C for 1 hour. The reaction mixture was quenched by adding water (50 mL) and extracted using DCM (50 mL x 3). The combined organic layers were washed with brine (10 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue which was purified by flash silica gel chromatography (from PE/EtOAc = 1/0 to 3/1, TLC: PE/EtOAc = 3/1, R _f = 0.43), tert -butyl N -[2-bromo-4-(prop-2-enoylamino)phenyl] -N -[[4-(trifluoromethyl)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] ⁺ .

Step 4 : tert -Butyl (4-acrylamido-2-(1-methyl-1 H -imidazol-4-yl)phenyl)(4-(trifluoromethyl)benzyl)carbamate

tert -Butyl N- [2-bromo-4-(prop-2-enoylamino)phenyl] -N -[[4-(trifluoromethyl)phenyl]methyl]carbamate in DMF (7 mL) Pd(dppf) in a solution of (200 mg, 400.54 μmol, 100%, 1 eq) and tributyl-(1-methylimidazol-4-yl)stanane (234.73 mg, 600.82 μmol, 95%, 1.5 eq). )Cl ₂ (29.31 mg, 40.05 μmol, 0.1 eq) was added. The mixture was stirred at 130° C. for 1 hour under N ₂ atmosphere. The reaction mixture was quenched by addition of water (50 mL) and extracted using EtOAc (50 mLx3). 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 purified by flash silica gel chromatography (from PE/EtOAc = 1/0 to 4/5, TLC: PE/EtOAc = 3/1, R _f = 0.20) to give tert -butyl N -[2-(1-methylimidazol-4-yl)-4-(prop- 2-enoylamino)phenyl]- N -[[4-(trifluoromethyl)phenyl]methyl]carbamate (100 mg, 199.80 μmol, 49.8% yield, 100.0% purity) was obtained as a yellow solid. 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] ⁺ .

Step 5 : N -(3-(1-methyl-1 H -imidazol-4-yl)-4-((4-(trifluoromethyl)benzyl)amino)phenyl)acrylamide

tert-Butyl N -[2-(1-methylimidazol-4-yl)-4-(prop-2-enoylamino)phenyl]- N -[[4-(tri) in DCM (3 mL) To a solution of fluoromethyl)phenyl]methyl]carbamate (100 mg, 199.80 μmol, 100%, 1 eq) 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 addition of water (30 mL) and extracted using EtOAc (30 mLx3). The combined organic layers were washed with brine (10 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue which was subjected to preparative HPLC (column: Agela DuraShell C18 150*25 mm*5 μm; mobile phase: purified with [water (0.05% NH ₃ H ₂ O+10 mM NH ₄ HCO ₃ )-ACN]; B%: 40%-70%, 10 min) and then lyophilized to form N -[ 3-(1-methylimidazol-4-yl)-4-[[4-(trifluoromethyl)phenyl]methylamino]phenyl]prop-2-enamide (10.48 mg, 26.17 μmol, 13.1% Yield, 100.0% purity) 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] ⁺ .

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) in 1,4-dioxane (10 mL) and H ₂ O (2 mL) Romethyl)-2-pyridyl]amino]benzenesulfonamide (600 mg, 1.13 mmol, 1 eq) and 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane To a solution of (348.48 mg, 2.26 mmol, 383.79 μL, 2 eq), Cs ₂ CO ₃ (737.21 mg, 2.26 mmol, 2 eq) and Pd(dppf)Cl ₂ (82.78 mg, 113.13 μmol, 0.1 eq) were added. . The mixture was stirred at 100° C. for 12 hours under N ₂ atmosphere. The reaction mixture was quenched by addition of water (50 mL) and extracted using EtOAc (50 mLx3). The combined organic layers were washed with brine (30 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue which was purified by flash silica gel chromatography (from 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, 91.6% yield, 98.9% purity) was obtained as a white solid. ^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] ⁺ .

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-benzenesulfone in DCM (2 mL) To a solution of amide (500 mg, 1.04 mmol, 98.9% purity, 1 eq) 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 addition of water (50 mL) and extracted using EtOAc (30 mLx3). 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 purified by flash silica gel chromatography (from PE/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, 77.7% yield, 95.9% purity) was obtained as a colorless oil. ^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 3 : 3-[(5S)-3-bromo-4,5-dihydroisoxazol-5-yl]- N -methyl-4-[[5-(trifluoromethyl)-2-pyridyl ]amino]benzenesulfonamide and 3-[(5R)-3-bromo-4,5-dihydroisoxazol-5-yl]- N -methyl-4-[[5-(trifluoromethyl)- 2-pyridyl]amino]benzenesulfonamide

N -methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]-3-vinyl-benzenesulfonamide (270 mg, 724.58 μmol, 95.9%, 1 eq) in EtOAc (10 mL) ) and NaHCO ₃ (608.70 mg, 7.25 mmol, 281.80 μL, 10 eq) was added to a stirred solution of dibromomethanone oxime (293.94 mg, 1.45 mmol, 2 eq). The reaction mixture was stirred at 25°C for 2 hours. The reaction mixture was quenched by adding water (50 mL) and extracted using 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 give 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%: purified with 40%-70%, 10 min) and then lyophilized to obtain a mixture This was carried out on a chiral SFC column: DAICEL CHIRALPAK IG (250 mm*30 mm, 10 um); Mobile phase: [0.1% NH ₃ ·H ₂ O MEOH]; B%: 40%-40%) to obtain 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) and lyophilized to give 3-[(5 S )-3-bromo-4,5- dihydroisoxazol-5-yl]- N -methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]benzenesulfonamide (44.51 mg, 92.87 μmol, 12.8% yield, 100.0% Purity, SFC: R _t = 1.658, ee = 100%, [α] ^31.4 _D = -44.4 (CH ₃ OH, c = 0.054 g/100 mL)) was obtained as a yellow solid. ^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] ⁺ . 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-[(5 R )-3-bromo-4,5- dihydroisoxazol-5-yl]- N -methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]benzenesulfonamide (48.81 mg, 101.84 μmol, 14.1% yield, 100.0% Purity, SFC: R _t = 1.982, ee = 100%, [α] ^31.4 _D = + 40.0 (CH ₃ OH, c = 0.050 g/100 mL)) was obtained as a yellow solid. ^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] ⁺ .

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 (25 mL) ) AcOH (50.39 mg, 839.16 μmol, 47.99 uL, 0.1 eq) was added to the solution. The mixture was stirred at 60°C for 4 hours. NaBH ₃ CN (2.64 g, 41.96 mmol, 5 eq) was added at 25°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 using EtOAc (100 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 give a residue which was purified by flash silica gel chromatography (from 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, 43.7% yield, 89.0% purity) was obtained as a yellow oil. ¹ H 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] ⁺ .

Step 2 : tert -Butyl N -(5-bromo-2-iodo-phenyl)- N -[[4-(trifluoromethyl)phenyl]methyl]carbamate

DMAP in a solution of 5-bromo-2-iodo- N -[[4-(trifluoromethyl)phenyl]methyl]aniline (1.88 g, 3.67 mmol, 89%, 1 eq) in THF (20 mL). (448.24 mg, 3.67 mmol, 1 eq) and Boc ₂ O (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 (from PE/EtOAc = 1/0 to 10/1, TLC: PE/EtOAc = 10/1, R _f = 0.8) to give 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, 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 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, 96.3% purity, 1 eq), tributyl-(1-methylimidazol-4-yl)stanane (662.53 mg, 1.73 mmol, 97% purity, 1 eq) and Pd(dppf)Cl ₂ (126.70 mg, The mixture (173.15 μmol, 0.1 eq) was degassed and purged three times with N ₂ and the mixture was stirred at 120° C. for 4 hours under N ₂ atmosphere. The reaction mixture was quenched by adding water (100 mL) and extracted using 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 purified by flash silica gel chromatography (from PE/EtOAc = 1/0 to 3/1, TLC: PE/EtOAc = 3/1, R _f = 0.40) to give tert -butyl N -[5-bromo-2-(1-methylimidazol-4-yl)phenyl] - N -[[4-(trifluoromethyl)phenyl]methyl]carbamate (530 mg, 934.66 μmol, 54.0% yield, 90.0% purity) was obtained as a dark brown oil. ^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 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 -[ in 1,4-dioxane (4.5 mL) and H ₂ O (1.5 mL) [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) The mixture was degassed and purged three times with N ₂ and the mixture was stirred at 100° C. for 12 hours under N ₂ atmosphere. The reaction mixture was quenched by adding water (50 mL) and extracted using EtOAc (30 mL x 3). The combined organic layers were washed with brine (10 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue which was purified by flash silica gel chromatography (from PE/EtOAc = 1/0 to 3/1, TLC: PE/EtOAc = 3/1, R _f = 0.40) to give tert -butyl N -[2-(1-methylimidazol-4-yl)-5-vinyl-phenyl]- N -[[4-(trifluoromethyl)phenyl]methyl]carbamate (219 mg, 464.34 μmol, 49.7% yield, 97.0% purity) was obtained as a yellow oil. ^1H NMR (400 MHz, CDCl ₃ ) 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] ⁺ .

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] in EtOAc (3 mL) NaHCO ₃ (301.02 mg, 3.58 mmol, 10 eq) was added to a solution of carbamate (169 mg, 358.33 μmol, 97%, 1 eq) and dibromomethanone oxime (109.02 mg, 537.49 μmol, 1.5 eq). . The mixture was stirred at 25°C for 4 hours. The reaction mixture was quenched by addition of water (50 mL) and extracted using EtOAc (30 mLx3). The combined organic layers were washed with brine (10 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure 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 yellow. Obtained as a solid, which was used in the next step without further purification. ^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 6 : ( S )-5-(3-Bromo-4,5-dihydroisoxazol-5-yl)-2-(1-methyl-1H-imidazol-4-yl)-N- ( 4 -(trifluoromethyl)benzyl)aniline and ( R )-5-(3-bromo-4,5-dihydroisoxazol-5-yl)-2-(1-methyl-1H-imidazole-4 -yl)- N -(4-(trifluoromethyl)benzyl)aniline

tert-Butyl N -[5-(3-bromo-4,5-dihydroisoxazol-5-yl)-2-(1-methylimidazol-4-yl)phenyl] in DCM (3 mL) - To a solution of N -[[4-(trifluoromethyl)phenyl]methyl]carbamate (248 mg, 428.02 μmol, 1 eq) 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 addition of water (100 mL) and extracted using EtOAc (60 mLx3). 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 purified by flash silica gel chromatography (in PE/EtOAc = 100/1 to 3). /1, TLC: PE/EtOAc = 3/1, R _f = 0.05) to obtain the 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 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%: 52% -82%, 10 minutes) and lyophilized to obtain ( S )-5-(3-bromo-4,5-dihydroisoxazol-5-yl)-2-(1-methyl-1H-imidazole. -4-yl)- N -(4-(trifluoromethyl)benzyl)aniline (10.44 mg, 21.35 μmol, 5.0% yield, 98.5% purity, 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. ^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] ⁺ . Peak 2 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%: 51% -81%, 10 minutes) and lyophilized to obtain ( R )-5-(3-bromo-4,5-dihydroisoxazol-5-yl)-2-(1-methyl-1H-imidazole. -4-yl)- N -(4-(trifluoromethyl)benzyl)aniline (9.5 mg, 18.99 μmol, 4.4% yield, 95.9% purity, SFC: R _t = 2.482, ee = 98.8%, ee = 98.8 %, [α] ^32.0 _D = -187.5 (MeOH, c = 0.016 g/100 mL)) was obtained as a white solid. ^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] ⁺ .

I-14

Step 1 : N -(2-bromo-4-nitrophenyl)-5-(trifluoromethyl)pyridin-2-amine

NaH (1.36 g, 34.09 mmol, 60%, 3 eq) was added to a solution of 5-(trifluoromethyl)pyridin-2-amine (1.84 g, 11.36 mmol, 1 eq) in THF (30 mL) at 0°C. Added. The mixture was stirred for 30 minutes. 2-Bromo-1-fluoro-4-nitro-benzene (2.5 g, 11.36 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 using 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 purified by flash silica gel chromatography (from PE/EtOAc = 100/1 to 10/1, TLC: PE/EtOAc = 5/1, R _f = 0.65) to obtain N -(2-bromo-4-nitro-phenyl)-5-(trifluoromethyl)pyridin-2-amine (850 mg, 2.00 mmol, 17.6% Yield, 85.0% purity) was obtained as a yellow solid. ¹ H 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] ⁺ .

Step 2 : 2-(1-methyl-1 H -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 trifluoromethylamine in DMF (15 mL) under N ₂ atmosphere. Pd(dppf)Cl ₂ (70.73 mg, 96.66 μmol, 0.05 eq) was added to a solution of butyl-(1-methylimidazol-4-yl)stanane (1.58 g, 3.87 mmol, 91%, 2 eq). did. The mixture was stirred at 130° C. for 12 hours under N ₂ 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 Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue which was purified by silica gel column chromatography (pure PE to PE/EtOAc). = 0/1, TLC: PE/EtOAc = 0/1, R _f = 0.34) to give 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. ^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 3 : N -(3-(1-methyl-1 H -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 in DCM (3 mL) at 0° C. under N ₂ atmosphere, Prop-2-enoyl chloride (58.46 mg, 645.88 μmol) in a solution of 4-diamine (230 mg, 496.83 μmol, 72%, 1 eq) and Et ₃ N (150.82 mg, 1.49 mmol, 207.46 μL, 3 eq). , 52.66 μL, 1.3 eq) was added. 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 give 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) purified to N -[3-( 1-methylimidazol-4-yl)-4-[[5-(trifluoromethyl)-2-pyridyl]amino]phenyl]prop-2-enamide (85.15 mg, 215.43 μmol, 43.4% Yield, 98.2% purity) was obtained as a white solid. ^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] ⁺ .

I-15 and I-16 (isomers of I-27)

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

3-(3-bromo-4,5-dihydroisoxazol-5-yl) -N -methyl-4-[[5-(trifluoromethyl)- in 1,4-dioxane (10 mL) To a solution of 2-pyridyl]amino]benzenesulfonamide (900 mg, 723.35 μmol, 38.5% purity, 1 eq) was added aqueous HCl (2 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 using EAOAc (50 mLx 3). The combined organic layers were washed with brine (50 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue which was subjected to preparative HPLC (column: Agela DuraShell C ₁₈ 150*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 obtain the mixture. This was obtained and purified by chiral SFC (column: DAICEL CHIRALPAK AS (250 mm*30 mm, 10 μm); mobile phase: [0.1% NH ₃ ·H ₂ O EtOH]; B%: 25%-20%) Separation gave peak 1 and peak 2. 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-[(5 S )-3-chloro-4,5- dihydroisoxazol-5-yl]- N -methyl-4-[[5-(trifluoromethyl)-2-pyridyl]amino]benzenesulfonamide (48.36 mg, 111.22 μmol, 15.4% yield, 100.0% Purity, SFC: R _t = 1.382, ee = 99.84%, [α] ^31.4 _D = - 46.15 (CH ₃ OH, c = 0.052 g/100 mL)) was obtained as a white solid. ^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-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). 1-(4-Methoxyphenyl)- N -methyl-methanamine (571.67 mg, 3.78 mmol, 1.1 eq) was added to the solution. The mixture was stirred at -30°C for 1 hour. TLC (PE/EtOAc = 3/1, R _f = 0.56) showed that the starting material was completely consumed and one new spot was formed. 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. ¹ H 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] ⁺ .

Step 2 : 6-Amino-5-bromo- N -[(4-methoxyphenyl)methyl] -N -methyl-pyridine-3-sulfonamide

5-Bromo-6-chloro- N -[(4-methoxyphenyl)methyl]- N -methyl-pyridine-3-sulfonamide (950 mg, 2.15 mmol, 92% purity, 1) in THF (5 mL) To the solution of eq) was added NH ₃ .H ₂ O (1.48 mL, 28% purity, 5 eq). The mixture was stirred at 100° C. under microwave for 12 hours. TLC (PE/EtOAc = 1/1, R _f = 0.31) showed that the starting material was completely consumed and one new spot was formed. The solvent was removed to obtain 6-amino-5-bromo- N -[(4-methoxyphenyl)methyl]- N -methyl-pyridine-3-sulfonamide (860 mg, 2.12 mmol, 98.2% yield, 95.0% purity). ) was obtained as a yellow solid, which was used in the next step without further purification. ^1H NMR (500 MHz, DMSO- d6 ) 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 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% pure, in anisole (20 mL) Pd(OAc) ₂ (42.24 mg, 188.15 μmol, 0.15 eq) in a solution of 1 eq) and 1-iodo-4-(trifluoromethyl)benzene (409.42 mg, 1.51 mmol, 221.31 μL, 1.2 eq); Xantphos (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 hours under N ₂ 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 (from PE/EtOAc = 1/0 to 3/1, TLC: PE/EtOAc = 3/1, Rf = 0.57). Purified to obtain 5-bromo- N -[(4-methoxyphenyl)methyl]- N -methyl-6-[4-(trifluoromethyl)anilino]pyridine-3-sulfonamide (587 mg, 1.05 mg) mmol, 83.8% yield, 95.0% purity) was obtained as a yellow solid. ^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 = 8.5 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 = 8.5 Hz, 2H), 4.14 (s, 2H), 3.81 ( s, 3H), 2.65 (s, 3H); ES-LCMS m/z 530.0, 532.0 [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-(trifluoro) in 1,4-dioxane (18 mL) and H ₂ O (3 mL) methyl)anilino]pyridine-3-sulfonamide (587 mg, 1.05 mmol, 95% purity, 1 eq) and 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxabo Add 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) to a solution of rolan (323.88 mg, 2.10 mmol, 356.70 μL, 2 eq). did. The mixture was stirred at 90°C for 16 hours under N ₂ 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 purified by flash silica gel chromatography (from PE/EtOAc = 1/0 to 3/1, TLC: PE/EtOAc = 3/1, R _f = 0.49 ) purified by N -[(4-methoxyphenyl)methyl]- N -methyl-6-[4-(trifluoromethyl)anilino]-5-vinyl-pyridine-3-sulfonamide (350 mg, 732.99 μmol, 69.7% yield, 100.0% purity) was obtained as a yellow solid. ^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 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) in DCM (5 mL) mg, 628.27 μmol, 100% purity, 1 eq) 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 that the starting material was completely consumed and one new spot was formed. The mixture was concentrated to obtain N -methyl-6-[4-(trifluoromethyl)anilino]-5-vinyl-pyridine-3-sulfonamide (330 mg, 616.07 μmol, 98.1% yield, 88.0% purity, TFA). was obtained 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 = 11.0 Hz, 1H), 2.59 (s, 3H).

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) in EtOAc (20 mL) ) and dibromomethanone oxime (249.92 mg, 1.23 mmol, 2 eq) were added NaHCO ₃ (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 that the starting material was completely consumed and one new spot was formed. The mixture was filtered and the filtrate was concentrated to give a residue which was purified by flash silica gel chromatography (from PE/EtOAc = 1/0 to 2/1, TLC: PE/EtOAc = 1/1, R _f = 0.78 ) Purified with 5-(3-bromo-4,5-dihydroisoxazol-5-yl)- N -methyl-6-[4-(trifluoromethyl)anilino]pyridine-3-sulfonamide (290 mg, 574.83 μmol, 93.3% yield, 95.0% purity) was obtained as a yellow solid. ^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 7 : ( S )-5-(3-Bromo-4,5-dihydroisoxazol-5-yl)-N-methyl-6-((4-(trifluoromethyl)phenyl)amino)pyridine -3-sulfonamide

Compound 5-(3-bromo-4,5-dihydroisoxazol-5-yl)- N -methyl-6-[4-(trifluoromethyl)anilino]pyridine-3-sulfonamide (100 mg , 198.22 μmol, 95% purity, 1 eq) was purified by chiral SFC (column: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 um); mobile phase: [0.1% NH ₃ H ₂ O EtOH]; B%: 30% -30%) to obtain peak 1 and peak 2. 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 lyophilized to give ( S )-5-(3-bromo-4,5-di Hydroisoxazol-5-yl)- N -methyl-6-((4-(trifluoromethyl)phenyl)amino)pyridine-3-sulfonamide (32.79 mg, 68.42 μmol, 34.5% yield, 100.0% purity, 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 ] ⁺ . 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 ( 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, 33.3% yield, 100.0 % Purity, 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] ⁺ .

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)stanane (2.72 g, 6.71 g) in DMF (20 mL) mmol, 91.5%, 1 eq) was added to the mixture of Pd(dppf)Cl ₂ (491.22 mg, 671.32 μmol, 0.1 eq). The mixture was stirred at 130° C. for 12 hours under N ₂ atmosphere. The mixture was diluted with water (100 mL) and extracted using EtOAc (100 mL x 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated in vacuo to give a residue which was purified by flash silica gel chromatography (from PE/EtOAc = 100/1). to 0/1, TLC: PE/EtOAc = 0/1, R _f = 0.30) to obtain 4-bromo-2-(1-methylimidazol-4-yl)aniline (1 g, 3.25 mmol, 48.5% yield, 82.0% purity) 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] ⁺ .

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) in 1,4-dioxane (30 mL) and H ₂ O (6 mL) eq) and 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (1.00 g, 6.51 mmol, 1.10 mL, ₂ eq). (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 hours under N ₂ atmosphere. The mixture was diluted with water (100 mL) and extracted using EtOAc (100 mL x 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated in vacuo to give a residue which was purified by flash silica gel chromatography (in PE/EtOAc = 100/1). to 0/1, TLC: PE/EtOAc = 0/1, R _f = 0.35) to obtain 2-(1-methylimidazol-4-yl)-4-vinyl-aniline (500 mg, 2.16 mmol, 66.4% yield, 86.0% purity) was obtained as a brown oil. ^1H NMR (400 MHz, DMSO- d6 ) 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 3 : 4-(3-Bromo-4,5-dihydroisoxazol-5-yl)-2-(1-methylimidazol-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/EtOAcR = 0/1, R _f = 0.20) to give 4-( 3-Bromo-4,5-dihydroisoxazol-5-yl)-2-(1-methylimidazol-4-yl)aniline (150 mg, 434.35 μmol, 20.1% yield, 93.0% purity) Obtained as a yellow oil. ^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 4 : 4-[(5 S )-3-bromo-4,5-dihydroisoxazol-5-yl]-2-(1-methylimidazol-4-yl) -N -[[4 -(trifluoromethyl)phenyl]methyl]aniline and 4-[(5 R )-3-bromo-4,5-dihydroisoxazol-5-yl]-2-(1-methylimidazole- 4-yl)- N -[[4-(trifluoromethyl)phenyl]methyl]aniline

4-(3-bromo-4,5-dihydroisoxazol-5-yl)-2-(1-methylimidazol-4-yl)aniline (120 mg, 347.48 μmol, 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) in a solution of 93.0%, 1 eq). , 2 eq) was added. The mixture was stirred at 25°C for 12 hours. The mixture was diluted with water (30 mL) and extracted using EtOAc (30 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 which was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm* 5 μm; Mobile phase: [Water (0.05% NH ₃ · H ₂ O+10 mM NH ₄ HCO ₃ )-ACN]; B%: 60%-90%, 10 min) and then lyophilized to obtain the product. Obtained. The product was separated by SFC (column: DAICEL CHIRALPAK IG (250 mm * 50 mm, 10 μm); mobile phase: [0.1% NH ₃ ·H ₂ O EtOH]; B%: 60%-60%) to obtain peaks 1 and Peak 2 was obtained. Peak 1 was concentrated under reduced pressure to give a residue, which was dissolved in MeCN (20 mL) and H ₂ O (40 mL) and lyophilized to give 4-[(5 S )-3-bromo-4,5- dihydroisoxazol-5-yl]-2-(1-methylimidazol-4-yl)- N -[[4-(trifluoromethyl)phenyl]methyl]aniline (24.61 mg, 50.72 μmol, 14.6 % yield, 98.8% purity, 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. ^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] ⁺ . Peak 2 was concentrated under reduced pressure to give a residue, which was dissolved in MeCN (20 mL) and H ₂ O (40 mL) and lyophilized to give 4-[(5 R )-3-bromo-4,5- dihydroisoxazol-5-yl]-2-(1-methylimidazol-4-yl)- N -[[4-(trifluoromethyl)phenyl]methyl]aniline (24.58 mg, 50.61 μmol, 14.6 % yield, 98.7% purity, 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. ^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] ⁺ .

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 in 1,4-dioxane (10 mL) ] To a solution of pyridine-3-sulfonamide (185 mg, 366.70 μmol, 95% purity, 1 eq), HCl (4 M, 0.5 mL) was added. 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 (20 mL x 2). 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 analyzed by preparative TLC (PE/EtOAc = 1/1, R _f = 0.71) to obtain the product, which was purified by chiral SFC (column: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 um); mobile phase: [0.1% NH ₃ H ₂ O EtOH]; B%: 35%- 35%) to obtain 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) and lyophilized to give ( S )-5-(3-chloro-4,5-dihydroyl Soxazol-5-yl)- N -methyl-6-((4-(trifluoromethyl)phenyl)amino)pyridine-3-sulfonamide (43.65 mg, 97.09 μmol, 26.5% yield, 96.7% purity, 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 = 5.3 Hz, 3H); ES-LCMS m/z 435.0 [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-sulfone amides

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, 60% purity, 4 eq) , the mixture was stirred at 0°C for 0.5 hours. Add 5-bromo-6-chloro- N -[(4-methoxyphenyl)methyl]- N -methyl-pyridine-3-sulfonamide (160 mg, 374.67 μmol, 95% purity, 1 eq), The mixture was stirred at 25°C for 3 hours. The reaction mixture was diluted with H ₂ O (20 mL) and extracted using 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. 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 rinsed with MeOH (3 mL x 2). The solid was collected, dried in vacuo, and 5-bromo- N -[(4-methoxyphenyl)methyl] -N -methyl-6-[[5-(trifluoromethyl)-2-pyridyl]amino. ]Pyridine-3-sulfonamide (160 mg, 301.12 μmol, 80.8% yield, 100.0% purity) was obtained as a white solid. ^1H NMR (400 MHz, DMSO- d6 ) 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] ⁺ .

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) in 1,4-dioxane (6 mL) and H ₂ O (1 mL) 4,4,5,5-tetramethyl-2-vinyl-1 in a solution of romethyl)-2-pyridyl]amino]pyridine-3-sulfonamide (260 mg, 489.32 μmol, 100% purity, 1 eq) ,3,2-dioxaborolane (301.45 mg, 1.96 mmol, 331.99 μL, 4 eq), Pd(dppf)Cl ₂ (35.80 mg, 48.93 μmol, 0.1 eq) and Cs ₂ CO ₃ (318.86 mg, 978.64 μmol) , 2 eq) was added. The mixture was stirred at 90° C. for 12 hours under N ₂ atmosphere. The reaction mixture was diluted with H ₂ O (20 mL) and extracted using 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 purified by flash silica gel chromatography (from PE/EtOAc = 1/0). to 5/1, TLC: PE/EtOAc = 5/1, R _f = 0.35) to give N -[(4-methoxyphenyl)methyl]- N -methyl-6-[[5-(trifluoro Methyl)-2-pyridyl]amino]-5-vinyl-pyridine-3-sulfonamide (200 mg, 409.62 μmol, 83.7% yield, 98.0% purity) was obtained as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 8.69 (d, J = 2.3 Hz, 1H), 8.64 (d, J = 8.8 Hz, 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 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- in DCM (3 mL) To a solution of 3-sulfonamide (200 mg, 409.62 μmol, 98% purity, 1 eq) 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 obtain N -methyl-6-[[5-(trifluoromethyl)-2-pyridyl]amino]-5-vinyl-pyridine-3-sulfonamide (140 mg, crude) as a yellow solid. Obtained. ^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 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-(trifluoro methyl)-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 to the solution. 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 purified by flash silica gel chromatography (from PE/EtOAc = 1/0 to 1/1, TLC: PE/EtOAc = 1/1, R _f = 0.32 ) to obtain the product, which 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-[(5 R )-3-bromo-4,5-dihydroisoxazol-5-yl] -N -methyl-6-[[5-(trifluoromethyl) -2-pyridyl]amino]pyridine-3-sulfonamide (39.4 mg, 81.48 μmol, 20.9% yield, 99.3% purity, 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] ⁺ .

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)cyclohexanamine (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, 100.0% purity, 1 eq) was added. The reaction mixture was stirred at 140°C for 3 hours under N ₂ atmosphere. The reaction mixture was diluted with H ₂ O (30 mL) and extracted using EtOAc (60 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 give a residue which was purified by flash silica gel chromatography (from PE/EtOAc = 100/1 to 3/1, TLC: PE/EtOAc = 3/1, R _f = 0.36) to obtain the crude product, which was subjected to preparative HPLC (column: Boston Green ODS 150*30 mm*5 μm; mobile phase: [Water (0.05% HCl)-ACN]; B%: 53%-83%, 10 min) to obtain 3-bromo- N -methyl-4-[[4-(trifluoromethyl)cyclo Hexyl]amino]benzenesulfonamide (250 mg, 571.92 μmol, 19.2% yield, 95.0% purity) was obtained as a white solid. ¹ H 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] ⁺ .

Step 2 : N -methyl-4-[[4-(trifluoromethyl)cyclohexyl]amino]-3-vinyl-benzenesulfonamide

3-Bromo- N -methyl-4-[[4-(trifluoromethyl)cyclohexyl]amino]benzenesulfonamide (210) in 1,4-dioxane (6 mL) and H ₂ O (2 mL) 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) Pd(dppf)Cl ₂ (32.31 mg, 44.16 μmol, 0.1 eq) and Cs ₂ CO ₃ (359.73 mg, 1.10 mmol, 2.5 eq) were added to the solution. The mixture was stirred at 100° C. for 2 hours under N ₂ atmosphere. The mixture was diluted with water (30 mL) and extracted using EtOAc (30 mL x 3). 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 purified by flash silica gel chromatography (from 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-benzene. Sulfonamide (160 mg, 435.75 μmol, 98.7% yield, 98.7% purity) was obtained as a yellow oil. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 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 3 : 3-[(5 S )-3-bromo-4,5-dihydroisoxazol-5-yl] -N -methyl-4-[[4-(trifluoromethyl)cyclohexyl]amino ]Benzenesulfonamide

To a solution of N -methyl-4-[[4-(trifluoromethyl)cyclohexyl]amino]-3-vinyl-benzenesulfonamide (110 mg, 297.45 μmol, 1 eq) in EtOAc (5 mL) dibro. Momethanone 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 one new spot was formed. The mixture was diluted with water (50 mL) and extracted using EtOAc (50 mL 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 purified by flash silica gel chromatography (from 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% NH _{3 •} H ₂ O ETOH]; B%: 25%-25%) to obtain peak 1 and peak 2. Peak 1 was concentrated under reduced pressure to give a residue, which was dissolved in MeCN (20 mL) and H ₂ O (20 mL) and lyophilized to give 3-[(5 S )-3-bromo-4,5- dihydroisoxazol-5-yl]- N -methyl-4-[[4-(trifluoromethyl)cyclohexyl]amino]benzenesulfonamide (60 mg, 122.40 μmol, 41.1% yield, 98.8% purity, 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, 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] ⁺ .

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 regulates cell proliferation and apoptosis. It is activated by high cell density and cellular stress, stopping cell proliferation and inducing apoptosis. The mammalian Hippo pathway includes MST kinase and LATS kinase. When the Hippo pathway is activated, MST kinase phosphorylates LATS kinase, which in turn phosphorylates the transcriptional coactivators YAP and TAZ. Unphosphorylated YAP and TAZ can translocate to the nucleus and interact with TEAD/TEF transcription factors, turning on cell cycle-promoting gene transcription. However, when phosphorylated, YAP and TAZ are mobilized from the nucleus to the cytoplasm, whereby YAP and TAZ dependent gene transcription is turned off. Dysfunction of the Hippo pathway is frequently detected in human cancer, and its downregulation is correlated with the aggressive nature of cancer cells and poor prognosis.

explanation

TEAD Reporter - The MCF7 cell line contains the firefly luciferase gene under the control of a TEAD response element stably integrated into the human breast cancer cell line MCF7. Inside the cell, basal unphosphorylated YAP/TAZ remains in the nucleus and induces constitutive expression of the luciferase reporter. The cell lines are validated for inhibition of luciferase reporter expression by activators of the Hippo pathway.

apply

Monitor Hippo pathway activity.

Select activators or inhibitors of the Hippo pathway.

form

Each vial contains ~1.5 X 106 cells in 1 ml of 10% DMSO.

save

Upon receipt, store in liquid nitrogen.

General culture conditions

Thaw 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), 1 mM MEM medium (Hyclone #SH30024.01) supplemented with sodium pyruvate (Hyclone #SH30239.01), 1% penicillin/streptomycin (Hyclone SV30010.01), and 10 μg/ml insulin (Sigma-Aldrich #I0516).

Growth medium 1B (BPS Bioscience #79531) + 10 μg/ml insulin (Sigma-Aldrich #I0516): Thaw medium 1 (BPS Cat. #60187) + 10 μg/ml insulin (Sigma-Aldrich #I0516), and Geneticin (Invitrogen #11811031) at 400 μg/ml.

Cells should be grown at 37°C with 5% CO ₂ using growth medium 1B containing 10 μg/ml insulin. It may be necessary to adjust the percentage of CO ₂ in the incubator depending on the level of NaHCO ₃ in the basal medium.

To thaw the cells, rapidly 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 It is recommended to resuspend in pre-warmed Thawing Medium 1 + Insulin (without Geneticin), transfer the resuspended cells to a T25 flask, and incubate in a CO ₂ incubator at 37°C overnight. The next day, replace the medium with fresh thawed medium 1 + insulin (no Geneticin) and continue to grow the medium in a CO ₂ incubator at 37°C until the cells are ready to divide. In the first passage, exchange growth medium 1B + 10 μg/ml insulin (thawing medium 1, containing insulin and geneticin). Cells must divide before reaching full confluence.

To passage cells, rinse the cells with phosphate-buffered saline (PBS) and detach the cells from the culture vessel using 0.25% trypsin/EDTA. Add growth medium 1B + 10 μg/ml insulin (thaw medium 1, containing insulin and geneticin), transfer to a tube, spin down the cells, then resuspend the cells, and add appropriate aliquots of the cell suspension. Inoculate a new culture vessel. Weekly subculture ratio: 1:5 to 1:10.

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

Functional verification and verification performance

The following assay is designed in 96 well format. To perform assays in various tissue culture formats, cell numbers and reagent volumes must be adjusted appropriately.

Substances needed for cell culture but not supplied

해동 배지 1(BPS Bioscience #60187) + 10 μg/ml의 인슐린

Thawing medium 1 (BPS Bioscience #60187) + 10 μg/ml insulin

성장 배지 1B(BPS Bioscience #79531) + 10 μg/ml의 인슐린

Growth medium 1B (BPS Bioscience #79531) + 10 μg/ml insulin

소 췌장으로부터의 인슐린 용액(Sigma-Aldrich #: I0516)

Insulin solution from bovine pancreas (Sigma-Aldrich #: I0516)

Materials required for cell assays but not supplied

H₂O₂: Hippo 경로의 활성화제(MST 키나제 활성화)

H ₂ O ₂ : Activator of Hippo pathway (MST Kinase activation)

인슐린

insulin

검정 배지: 해동 배지 1(BPS. Cat. #: 60187) + 10 μg/ml의 인슐린

Assay medium: Thaw medium 1 (BPS. Cat. #: 60187) + 10 μg/ml insulin

소 췌장으로부터의 인슐린 용액(Sigma-Aldrich Cat. #: I0516)

Insulin solution from bovine pancreas (Sigma-Aldrich Cat. #: I0516)

오카다산(BPS bioscience #27047): Hippo 경로의 활성화제(MST 키나제 활성화). DMSO에서 10 mM 원액을 제조한다.

Okadasan (BPS bioscience #27047): Activator of the Hippo pathway (MST kinase activation). Prepare a 10 mM stock solution in DMSO.

96-웰 조직 배양 플레이트 또는 96-웰 조직 배양 처리된 백색의 투명 바닥 검정 플레이트

96-well tissue culture plate or 96-well tissue culture treated white, clear bottom assay plate

ONE-Step™ Luciferase Assay System(BPS, Cat. #60690)

ONE-Step™ Luciferase Assay System (BPS, Cat. #60690)

조도계

light meter

Mycoplasma Testing

To confirm the absence of Mycoplasma species, cell lines were screened using the PCR-based VenorGeM Mycoplasma Detection kit (Sigma-Aldrich).

TEAD reporter - Inhibition of TEAD reporter activity by Hippo pathway activator in MCF7 cells

1) Harvest TEAD reporter - MCF7 cells from the culture in growth medium, and inoculate the cells at a density of 35,000 cells per well into a white, transparent bottom 96-well microplate in 45 μl of assay medium.

2) Incubate the cells in a CO ₂ incubator at 37°C overnight.

3) Dilute the activator (H ₂ O ₂ or Okadasan) stock in assay medium. Add 5 μl of diluted activator to the well. The final concentration of DMSO in the assay medium is 0.1%.

4) Add 5 μl of assay medium containing the same concentration of DMSO without activator to the control wells.

5) Add 50 μl of assay medium containing DMSO to the cell-free control wells (to determine background luminescence).

6) Set to perform each treatment at least three times.

7) Incubate the cells in a CO ₂ incubator at 37°C for 5-6 hours.

8) Perform luciferase assay using the ONE-Step™ Luciferase Assay System according to the protocol provided below: Add 100 μl of ONE-Step™ Luciferase reagent per well and shake for ~15 minutes at room temperature. Measure luminescence using a illuminometer.

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 and in H226 and H28. The data is listed in Table 2 below. A: EC50 <0.1 uM; B : 0.1 uM ≤ EC50 ≤ 0.5 uM; C : EC50 > 0.5 uM.

In vitro data for certain exemplary compounds.

Example 3: Mouse pharmacokinetic study

Formulated compounds are administered intravenously or orally via gavage to BALB/c mice. Typically, blood is collected at 0.167, 0.5, 1, 2, 4, 6, 12, and 24 hours post-dose, processed into plasma by centrifugation, and stored at -80°C until analysis. An internal standard is added to each sample prior to protein precipitation using acetonitrile or TCA. The precipitate is filtered through a filter plate and the sample is analyzed by LC/MS/MS. Typically, a standard curve is prepared in plasma from 1.0 ng/mL to 3000 ng/mL and processed in the same manner as the samples. Sample analysis is typically performed using a suitable column equipped with an analytical UPLC column and provided with compounds eluted from the analytical column in a gradient from 30-95% 0.1% formic acid (v/v) in ACN:0.1% formic acid (v/v) in water. Performed on an LC/MS/MS system. Mass spectrometric detection of test compounds and internal standards is performed by MRM in positive mode. The pharmacokinetics of each compound were analyzed using Phoenix WinNonlin software (Pharsight, St. Louis, Mo) using noncompartmental analysis.

Example 4. CTGF data analysis

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

RNA is extracted from tumors using QIAZOL (Qiagen) lysis reagent, and then tissue is homogenized for 10 minutes using TissueLyser II (Qiagen). Once sample disruption and digestion are 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 MagMAX Mirvana Total RNA Isolation Kit. For RNA extraction, follow the manufacturer's recommended protocol for high-throughput isolation of RNA from tissue samples.

Expression of YAP/TEAD-regulated genes, CCN2 , encoding connective tissue growth factor (CTFG), and the housekeeping gene human glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were determined using TaqMan Gene Expression Master Mix and TaqMan probes. Quantify by qRT-PCR analysis. CTGF and GAPDH cycle threshold (Ct) values are determined for tumor cDNA samples, and CTGF expression is normalized to the internal control, GAPDH.

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

Example 5. Antiproliferation Assay

Individual cell lines are grown in media according to supplier instructions and seeded in 96-well plates at a density that ensures logarithmic growth for 72-96 hours. TEAD inhibitor compounds are administered to cells at a peak concentration of 10 μM, followed by 10-point three-fold serial dilutions. After 72-96 hours, proliferation is quantified using Cell TITERGLO ^™ (Promega, Inc.) and compared to vehicle control. Generate IC50 and EC50 values using Prism or XLFit curve fitting software.

Example 6. Inhibition of tumor growth in vivo

NCI-H226 In Vivo Efficacy Study

6-8 week old nu/nu nude mice (CRL) are inoculated subcutaneously with ^5x106 NCI-H226 human mesothelioma tumor cells on the right flank. Tumor growth is monitored twice per week using vernier calipers, and the mean tumor volume (MTV) is calculated using the formula V = W2 x L/2.

Once MTV reached approximately 150-200 mm ³ , animals were randomized into treatment groups (n = 8-10/group) and incubated with vehicle (5% DMSO + 95% PEG 400) or TEAD inhibitor for 27-40 days. Administered orally (PO) on a once-daily (QD) schedule.

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

%TGI= 100 - [MTV Processed/MTV Contrast] x 100

Tumor growth and body weight changes are measured twice per week.

For comparisons between vehicle control and TEAD inhibitor treatment groups, independent samples t-test is used for statistical analysis.

MSTO-211H in vivo efficacy study

6-8 week old SCID mice (CRL) are inoculated subcutaneously with 5 x 10 ⁶ MSTO-211H human mesothelioma tumor cells in the right flank. Tumor growth is monitored twice per week using vernier calipers, and the mean tumor volume (MTV) is calculated using the formula V = W2 x L/2.

When MTV reached approximately 150-200 mm ³ , animals were randomized into treatment groups (n = 6-8/group) and treated with vehicle (5% DMSO + 95% PEG 400) or TEAD inhibitor for 22-25 days. Administered orally (PO) once daily (QD).

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

%TGI= 100 - [MTV Processed/MTV Contrast] x 100

Tumor growth and body weight changes are measured twice per week.

For comparisons between vehicle control and TEAD inhibitor treatment groups, independent samples t-test is used for statistical analysis.

Example 7. TEAD selectivity assay

The TEAD targeting selectivity profile of the TEAD inhibitor compounds described herein can be compared to TEAD isoforms or variants, 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 any examples provided herein designed to monitor the interaction of YAP1 or TAZ. Co-immunoprecipitation techniques can be used to monitor protein-protein interactions, but it is difficult to increase the throughput based on the basic methodology required. Therefore, alternative but complementary assays can be used to detect different TEAD isoforms. Types or variants, 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 YAP1 (or TAZ).

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). Compounds are incubated with individual TEAD isoform proteins, fluorescent peptides, and potency is determined by quantifying the displacement of the peptides.

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, incorporated herein by reference in its entirety). . Briefly, the YAP-binding domain of each TEAD isoform is transiently co-expressed with the TEAD-binding domain, or YAP1 or TAZ, in HEK293 cells, 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 hereby incorporated 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 the fluorescence polarization assay, these chimeric gene fusions are recombinantly expressed in bacterial or insect cells and used as in vitro biochemical assays using a luciferase readout similar to cell-based assays.

Another exemplary assay is Karatas et al., “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 turn-on probe to a cysteine in the central pocket of a TEAD isoform by a small molecule, as described in 2020, the entire text of which is incorporated herein by reference. is incorporated into. Briefly, a thiol-reactive pro-fluorescent probe, N-(4-(7-diethylamino-4-methylcoumarin-3-yl)phenyl)maleimide (CPM) is used. The fluorescence in CPM is quenched due to the maleimide substitution on the phenyl group, which modulates the resonance between the coumarin carbonyl and the 7-amino group. However, when reacted with thiol, CPM fluorescence increases significantly. CPM can be used to probe TEAD inhibition because the reaction of free cysteine residues within the TEAD central pocket generates a fluorescent signal, and small molecules that bind to the TEAD central pocket with significant efficacy prevent covalent labeling of the cysteine by CPM. . Therefore, in the thiol conjugation assay, freshly prepared TEAD protein solution is added and incubated with test compounds in assay buffer at room temperature, then CPM solution is added and fluorescence is measured (Ex/Em: 380/470 nm) . Any test compound that binds to the TEAD central pocket with significant potency exhibits lower fluorescence or fluorescence inhibition compared to compounds that do not bind to the TEAD central pocket.

Example 8. Inhibition of Malignant Mesothelioma Tumor Cell Growth

The tumor cell growth inhibitory activity of the TEAD inhibitors described herein is assessed in the NCI-H2052 mesothelioma cell line with an NF2 mutation. These cell lines are selected based, in part, on their mutation status and the ability of siRNA directed against YAP, TAZ, or TEAD1-TEAD4 to inhibit cell proliferation. We also consider the nuclear localization of YAP upon confluence. 10,000 cells/well are plated in 96-well black plates with clear, flat-bottomed TC-treated imaging plates in regular medium with serum, which is replaced the next day with starvation medium containing 1% serum. . After growing in starvation medium for 1 day, cells are incubated with TEAD inhibitor compounds. The starting concentration is 30 μM, and serial dilutions in DMSO and medium are performed up to 0.1 μM to achieve a final DMSO concentration of 0.5%. Cells are then grown for 3 days, then EdU (Invitrogen, Molecular Probe) is added to each well to 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. Afterwards, the plate was incubated at room temperature for 15 minutes, washed twice with PBS, and 100 μl of Triton-100 containing 0.3% BSA was added per well to permeabilize the cells. After 20 min, wash the cells with PBS and perform EdU detection according to the manufacturer's instructions. For example, image acquisition is performed using ImageXpress Micro and analysis is performed using MetaXpress software (Molecular Device).

Although a number of embodiments of the invention have been described, it will be apparent that the basic example may be modified to provide other embodiments using the compounds and methods of the invention. Accordingly, it will be appreciated that the scope of the invention should be defined by the application and appended claims, and not by the specific embodiments presented by way of example.

SEQUENCE LISTING <110> IKENA ONCOLOGY, INC. <120> TEAD INHIBITORS AND USES THEREOF <130> 396661-017WO (187446) <140> PCT/US2021/072684 <141> 2021-12-02 <150> 63/120,441 <151> 2020-12-02 <160> 9 <170> KoPatentIn 3.0 <210> 1 <211> 426 <212> PRT <213> Homo sapiens <400> 1 Met Glu Pro Ser Ser Trp Ser Gly Ser Glu Ser Pro Ala Glu Asn Met 1 5 10 15 Glu Arg Met Ser Asp Ser Ala Asp Lys Pro Ile Asp Asn Asp Ala Glu 20 25 30 Gly Val Trp Ser Pro Asp Ile Glu Gln Ser Phe Gln Glu Ala Leu Ala 35 40 45 Ile Tyr Pro Pro Cys Gly Arg Arg Lys Ile Ile Leu Ser Asp Glu Gly 50 55 60 Lys Met Tyr Gly Arg Asn Glu Leu Ile Ala Arg Tyr Ile Lys Leu Arg 65 70 75 80 Thr Gly Lys Thr Arg Thr Arg Lys Gln Val Ser Ser His Ile Gln Val 85 90 95 Leu Ala Arg Arg Lys Ser Arg Asp Phe His Ser Lys Leu Lys Asp Gln 100 105 110 Thr Ala Lys Asp Lys Ala Leu Gln His Met Ala Ala Met Ser Ser Ala 115 120 125 Gln Ile Val Ser Ala Thr Ala Ile His Asn Lys Leu Gly Leu Pro Gly 130 135 140 Ile Pro Arg Pro Thr Phe Pro Gly Ala Pro Gly Phe Trp Pro Gly Met 145 150 155 160 Ile Gln Thr Gly Gln Pro Gly Ser Ser Gln Asp Val Lys Pro Phe Val 165 170 175 Gln Gln Ala Tyr Pro Ile Gln Pro Ala Val Thr Ala Pro Ile Pro Gly 180 185 190 Phe Glu Pro Ala Ser Ala Pro Ala Pro Ser Val Pro Ala Trp Gln Gly 195 200 205 Arg Ser Ile Gly Thr Thr Lys Leu Arg Leu Val Glu Phe Ser Ala Phe 210 215 220 Leu Glu Gln Gln Arg Asp Pro Asp Ser Tyr Asn Lys His Leu Phe Val 225 230 235 240 His Ile Gly His Ala Asn His Ser Tyr Ser Asp Pro Leu Leu Glu Ser 245 250 255 Val Asp Ile Arg Gln Ile Tyr Asp Lys Phe Pro Glu Lys Lys Gly Gly 260 265 270 Leu Lys Glu Leu Phe Gly Lys Gly Pro Gln Asn Ala Phe Phe Leu Val 275 280 285 Lys Phe Trp Ala Asp Leu Asn Cys Asn Ile Gln Asp Asp Ala Gly Ala 290 295 300 Phe Tyr Gly Val Thr Ser Gln Tyr Glu Ser Ser Glu Asn Met Thr Val 305 310 315 320 Thr Cys Ser Thr Lys Val Cys Ser Phe Gly Lys Gln Val Val Glu Lys 325 330 335 Val Glu Thr Glu Tyr Ala Arg Phe Glu Asn Gly Arg Phe Val Tyr Arg 340 345 350 Ile Asn Arg Ser Pro Met Cys Glu Tyr Met Ile Asn Phe Ile His Lys 355 360 365 Leu Lys His Leu Pro Glu Lys Tyr Met Met Asn Ser Val Leu Glu Asn 370 375 380 Phe Thr Ile Leu Leu Val Val Thr Asn Arg Asp Thr Gln Glu Thr Leu 385 390 395 400 Leu Cys Met Ala Cys Val Phe Glu Val Ser Asn Ser Glu His Gly Ala 405 410 415 Gln His His Ile Tyr Arg Leu Val Lys Asp 420 425 <210> 2 <211> 447 <212> PRT <213> Homo sapiens <400> 2 Met Gly Glu Pro Arg Ala Gly Ala Ala Leu Asp Asp Gly Ser Gly Trp 1 5 10 15 Thr Gly Ser Glu Glu Gly Ser Glu Glu Gly Thr Gly Gly Ser Glu Gly 20 25 30 Ala Gly Gly Asp Gly Gly Pro Asp Ala Glu Gly Val Trp Ser Pro Asp 35 40 45 Ile Glu Gln Ser Phe Gln Glu Ala Leu Ala Ile Tyr Pro Pro Cys Gly 50 55 60 Arg Arg Lys Ile Ile Leu Ser Asp Glu Gly Lys Met Tyr Gly Arg Asn 65 70 75 80 Glu Leu Ile Ala Arg Tyr Ile Lys Leu Arg Thr Gly Lys Thr Arg Thr 85 90 95 Arg Lys Gln Val Ser His Ile Gln Val Leu Ala Arg Arg Lys Ser 100 105 110 Arg Glu Ile Gln Ser Lys Leu Lys Asp Gln Val Ser Lys Asp Lys Ala 115 120 125 Phe Gln Thr Met Ala Thr Met Ser Ser Ala Gln Leu Ile Ser Ala Pro 130 135 140 Ser Leu Gln Ala Lys Leu Gly Pro Thr Gly Pro Gln Ala Ser Glu Leu 145 150 155 160 Phe Gln Phe Trp Ser Gly Gly Ser Gly Pro Pro Trp Asn Val Pro Asp 165 170 175 Val Lys Pro Phe Ser Gln Thr Pro Phe Thr Leu Ser Leu Thr Pro Pro 180 185 190 Ser Thr Asp Leu Pro Gly Tyr Glu Pro Pro Gln Ala Leu Ser Pro Leu 195 200 205 Pro Pro Pro Thr Pro Ser Pro Pro Pro Ala Trp Gln Ala Arg Gly Leu Gly 210 215 220 Thr Ala Arg Leu Gln Leu Val Glu Phe Ser Ala Phe Val Glu Pro Pro Pro 225 230 235 240 Asp Ala Val Asp Ser Tyr Gln Arg His Leu Phe Val His Ile Ser Gln 245 250 255 His Cys Pro Ser Pro Gly Ala Pro Pro Leu Glu Ser Val Asp Val Arg 260 265 270 Gln Ile Tyr Asp Lys Phe Pro Glu Lys Lys Gly Gly Leu Arg Glu Leu 275 280 285 Tyr Asp Arg Gly Pro Pro His Ala Phe Phe Leu Val Lys Phe Trp Ala 290 295 300 Asp Leu Asn Trp Gly Pro Ser Gly Glu Glu Ala Gly Ala Gly Gly Ser 305 310 315 320 Ile Ser Ser Gly Gly Phe Tyr Gly Val Ser Ser Gln Tyr Glu Ser Leu 325 330 335 Glu His Met Thr Leu Thr Cys Ser Ser Lys Val Cys Ser Phe Gly Lys 340 345 350 Gln Val Val Glu Lys Val Glu Thr Glu Arg Ala Gln Leu Glu Asp Gly 355 360 365 Arg Phe Val Tyr Arg Leu Leu Arg Ser Pro Met Cys Glu Tyr Leu Val 370 375 380 Asn Phe Leu His Lys Leu Arg Gln Leu Pro Glu Arg Tyr Met Met Asn 385 390 395 400 Ser Val Leu Glu Asn Phe Thr Ile Leu Gln Val Val Thr Asn Arg Asp 405 410 415 Thr Gln Glu Leu Leu Leu Cys Thr Ala Tyr Val Phe Glu Val Ser Thr 420 425 430 Ser Glu Arg Gly Ala Gln His His Ile Tyr Arg Leu Val Arg Asp 435 440 445 <210> 3 <211> 435 <212> PRT <213> Homo sapiens <400> 3 Met Ala Ser Asn Ser Trp Asn Ala Ser Ser Ser Pro Gly Glu Ala Arg 1 5 10 15 Glu Asp Gly Pro Glu Gly Leu Asp Lys Gly Leu Asp Asn Asp Ala Glu 20 25 30 Gly Val Trp Ser Pro Asp Ile Glu Gln Ser Phe Gln Glu Ala Leu Ala 35 40 45 Ile Tyr Pro Pro Cys Gly Arg Arg Lys Ile Ile Leu Ser Asp Glu Gly 50 55 60 Lys Met Tyr Gly Arg Asn Glu Leu Ile Ala Arg Tyr Ile Lys Leu Arg 65 70 75 80 Thr Gly Lys Thr Arg Thr Arg Lys Gln Val Ser Ser His Ile Gln Val 85 90 95 Leu Ala Arg Lys Lys Val Arg Glu Tyr Gln Val Gly Ile Lys Ala Met 100 105 110 Asn Leu Asp Gln Val Ser Lys Asp Lys Ala Leu Gln Ser Met Ala Ser 115 120 125 Met Ser Ser Ala Gln Ile Val Ser Ala Ser Val Leu Gln Asn Lys Phe 130 135 140 Ser Pro Pro Ser Pro Leu Pro Gln Ala Val Phe Ser Thr Ser Ser Arg 145 150 155 160 Phe Trp Ser Ser Pro Pro Leu Leu Gly Gln Gln Pro Gly Pro Ser Gln 165 170 175 Asp Ile Lys Pro Phe Ala Gln Pro Ala Tyr Pro Ile Gln Pro Pro Leu 180 185 190 Pro Pro Thr Leu Ser Ser Tyr Glu Pro Leu Ala Pro Leu Pro Ser Ala 195 200 205 Ala Ala Ser Val Pro Val Trp Gln Asp Arg Thr Ile Ala Ser Ser Arg 210 215 220 Leu Arg Leu Leu Glu Tyr Ser Ala Phe Met Glu Val Gln Arg Asp Pro 225 230 235 240 Asp Thr Tyr Ser Lys His Leu Phe Val His Ile Gly Gln Thr Asn Pro 245 250 255 Ala Phe Ser Asp Pro Pro Leu Glu Ala Val Asp Val Arg Gln Ile Tyr 260 265 270 Asp Lys Phe Pro Glu Lys Lys Gly Gly Leu Lys Glu Leu Tyr Glu Lys 275 280 285 Gly Pro Pro Asn Ala Phe Phe Leu Val Lys Phe Trp Ala Asp Leu Asn 290 295 300 Ser Thr Ile Gln Glu Gly Pro Gly Ala Phe Tyr Gly Val Ser Ser Gln 305 310 315 320 Tyr Ser Ser Ala Asp Ser Met Thr Ile Ser Val Ser Thr Lys Val Cys 325 330 335 Ser Phe Gly Lys Gln Val Val Glu Lys Val Glu Thr Glu Tyr Ala Arg 340 345 350 Leu Glu Asn Gly Arg Phe Val Tyr Arg Ile His Arg Ser Pro Met Cys 355 360 365 Glu Tyr Met Ile Asn Phe Ile His Lys Leu Lys His Leu Pro Glu Lys 370 375 380 Tyr Met Met Asn Ser Val Leu Glu Asn Phe Thr Ile Leu Gln Val Val 385 390 395 400 Thr Ser Arg Asp Ser Gln Glu Thr Leu Leu Val Ile Ala Phe Val Phe 405 410 415 Glu Val Ser Thr Ser Glu His Gly Ala Gln His His Val Tyr Lys Leu 420 425 430 Val Lys Asp 435 <210> 4 <211> 434 < 212> PRT <213> Homo sapiens <400> 4 Met Glu Gly Thr Ala Gly Thr Ile Thr Ser Asn Glu Trp Ser Ser Pro 1 5 10 15 Thr Ser Pro Glu Gly Ser Thr Ala Ser Gly Gly Ser Gln Ala Leu Asp 20 25 30 Lys Pro Ile Asp Asn Asp Ala Glu Gly Val Trp Ser Pro Asp Ile Glu 35 40 45 Gln Ser Phe Gln Glu Ala Leu Ala Ile Tyr Pro Pro Cys Gly Arg Arg 50 55 60 Lys Ile Ile Leu Ser Asp Glu Gly Lys Met Tyr Gly Arg Asn Glu Leu 65 70 75 80 Ile Ala Arg Tyr Ile Lys Leu Arg Thr Gly Lys Thr Arg Thr Arg Lys 85 90 95 Gln Val Ser Ser His Ile Gln Val Leu Ala Arg Arg Lys Ala Arg Glu 100 105 110 Ile Gln Ala Lys Leu Lys Asp Gln Ala Ala Lys Asp Lys Ala Leu Gln 115 120 125 Ser Met Ala Ala Met Ser Ser Ala Gln Ile Ile Ser Ala Thr Ala Phe 130 135 140 His Ser Ser Met Ala Leu Ala Arg Gly Pro Gly Arg Pro Ala Val Ser 145 150 155 160 Gly Phe Trp Gln Gly Ala Leu Pro Gly Gln Ala Gly Thr Ser His Asp 165 170 175 Val Lys Pro Phe Ser Gln Gln Thr Tyr Ala Val Gln Pro Pro Leu Pro 180 185 190 Leu Pro Gly Phe Glu Ser Pro Ala Gly Pro Ala Pro Ser Pro Ser Ala 195 200 205 Pro Pro Ala Pro Pro Trp Gln Gly Arg Ser Val Ala Ser Ser Lys Leu 210 215 220 Trp Met Leu Glu Phe Ser Ala Phe Leu Glu Gln Gln Gln Asp Pro Asp 225 230 235 240 Thr Tyr Asn Lys His Leu Phe Val His Ile Gly Gln Ser Ser Pro Ser 245 250 255 Tyr Ser Asp Pro Tyr Leu Glu Ala Val Asp Ile Arg Gln Ile Tyr Asp 260 265 270 Lys Phe Pro Glu Lys Lys Gly Gly Leu Lys Asp Leu Phe Glu Arg Gly 275 280 285 Pro Ser Asn Ala Phe Phe Leu Val Lys Phe Trp Ala Asp Leu Asn Thr 290 295 300 Asn Ile Glu Asp Glu Gly Ser Ser Phe Tyr Gly Val Ser Ser Gln Tyr 305 310 315 320 Glu Ser Pro Glu Asn Met Ile Ile Thr Cys Ser Thr Lys Val Cys Ser 325 330 335 Phe Gly Lys Gln Val Val Glu Lys Val Glu Thr Glu Tyr Ala Arg Tyr 340 345 350 Glu Asn Gly His Tyr Ser Tyr Arg Ile His Arg Ser Pro Leu Cys Glu 355 360 365 Tyr Met Ile Asn Phe Ile His Lys Leu Lys His Leu Pro Glu Lys Tyr 370 375 380 Met Met Asn Ser Val Leu Glu Asn Phe Thr Ile Leu Gln Val Val Thr 385 390 395 400 Asn Arg Asp Thr Gln Glu Thr Leu Leu Cys Ile Ala Tyr Val Phe Glu 405 410 415 Val Ser Ala Ser Glu His Gly Ala Gln His His Ile Tyr Arg Leu Val 420 425 430 Lys Glu <210> 5 <211> 221 <212> PRT <213> Homo sapiens <400> 5 Trp Gln Gly Arg Ser Ile Gly Thr Thr Lys Leu Arg Leu Val Glu Phe 1 5 10 15 Ser Ala Phe Leu Glu Gln Gln Arg Asp Pro Asp Ser Tyr Asn Lys His 20 25 30 Leu Phe Val His Ile Gly His Ala Asn His Ser Tyr Ser Asp Pro Leu 35 40 45 Leu Glu Ser Val Asp Ile Arg Gln Ile Tyr Asp Lys Phe Pro Glu Lys 50 55 60 Lys Gly Gly Leu Lys Glu Leu Phe Gly Lys Gly Pro Gln Asn Ala Phe 65 70 75 80 Phe Leu Val Lys Phe Trp Ala Asp Leu Asn Cys Asn Ile Gln Asp Asp 85 90 95 Ala Gly Ala Phe Tyr Gly Val Thr Ser Gln Tyr Glu Ser Ser Glu Asn 100 105 110 Met Thr Val Thr Cys Ser Thr Lys Val Cys Ser Phe Gly Lys Gln Val 115 120 125 Val Glu Lys Val Glu Thr Glu Tyr Ala Arg Phe Glu Asn Gly Arg Phe 130 135 140 Val Tyr Arg Ile Asn Arg Ser Pro Met Cys Glu Tyr Met Ile Asn Phe 145 150 155 160 Ile His Lys Leu Lys His Leu Pro Glu Lys Tyr Met Met Asn Ser Val 165 170 175 Leu Glu Asn Phe Thr Ile Leu Leu Val Val Thr Asn Arg Asp Thr Gln 180 185 190 Glu Thr Leu Leu Cys Met Ala Cys Val Phe Glu Val Ser Asn Ser Glu 195 200 205 His Gly Ala Gln His His Ile Tyr Arg Leu Val Lys Asp 210 215 220 <210> 6 <211> 230 <212> PRT <213> Homo sapiens <400> 6 Trp Gln Ala Arg Gly Leu Gly Thr Ala Arg Leu Gln Leu Val Glu Phe 1 5 10 15 Ser Ala Phe Val Glu Pro Pro Asp Ala Val Asp Ser Tyr Gln Arg His 20 25 30 Leu Phe Val His Ile Ser Gln His Cys Pro Ser Pro Gly Ala Pro Pro 35 40 45 Leu Glu Ser Val Asp Val Arg Gln Ile Tyr Asp Lys Phe Pro Glu Lys 50 55 60 Lys Gly Gly Leu Arg Glu Leu Tyr Asp Arg Gly Pro Pro His Ala Phe 65 70 75 80 Phe Leu Val Lys Phe Trp Ala Asp Leu Asn Trp Gly Pro Ser Gly Glu 85 90 95 Glu Ala Gly Ala Gly Gly Ser Ile Ser Ser Gly Gly Phe Tyr Gly Val 100 105 110 Ser Ser Gln Tyr Glu Ser Leu Glu His Met Thr Leu Thr Cys Ser Ser 115 120 125 Lys Val Cys Ser Phe Gly Lys Gln Val Val Glu Lys Val Glu Thr Glu 130 135 140 Arg Ala Gln Leu Glu Asp Gly Arg Phe Val Tyr Arg Leu Leu Arg Ser 145 150 155 160 Pro Met Cys Glu Tyr Leu Val Asn Phe Leu His Lys Leu Arg Gln Leu 165 170 175 Pro Glu Arg Tyr Met Met Asn Ser Val Leu Glu Asn Phe Thr Ile Leu 180 185 190 Gln Val Val Thr Asn Arg Asp Thr Gln Glu Leu Leu Leu Cys Thr Ala 195 200 205 Tyr Val Phe Glu Val Ser Thr Ser Glu Arg Gly Ala Gln His His Ile 210 215 220 Tyr Arg Leu Val Arg Asp 225 230 <210> 7 <211> 221 <212> PRT <213> Homo sapiens <400> 7 Trp Gln Asp Arg Thr Ile Ala Ser Ser Arg Leu Arg Leu Leu Glu Tyr 1 5 10 15 Ser Ala Phe Met Glu Val Gln Arg Asp Pro Asp Thr Tyr Ser Lys His 20 25 30 Leu Phe Val His Ile Gly Gln Thr Asn Pro Ala Phe Ser Asp Pro Pro 35 40 45 Leu Glu Ala Val Asp Val Arg Gln Ile Tyr Asp Lys Phe Pro Glu Lys 50 55 60 Lys Gly Gly Leu Lys Glu Leu Tyr Glu Lys Gly Pro Pro Asn Ala Phe 65 70 75 80 Phe Leu Val Lys Phe Trp Ala Asp Leu Asn Ser Thr Ile Gln Glu Gly 85 90 95 Pro Gly Ala Phe Tyr Gly Val Ser Ser Gln Tyr Ser Ser Ser Ala Asp Ser 100 105 110 Met Thr Ile Ser Val Ser Thr Lys Val Cys Ser Phe Gly Lys Gln Val 115 120 125 Val Glu Lys Val Glu Thr Glu Tyr Ala Arg Leu Glu Asn Gly Arg Phe 130 135 140 Val Tyr Arg Ile His Arg Ser Pro Met Cys Glu Tyr Met Ile Asn Phe 145 150 155 160 Ile His Lys Leu Lys His Leu Pro Glu Lys Tyr Met Met Asn Ser Val 165 170 175 Leu Glu Asn Phe Thr Ile Leu Gln Val Val Thr Ser Ser Arg Asp Ser Gln 180 185 190 Glu Thr Leu Leu Val Ile Ala Phe Val Phe Glu Val Ser Thr Ser Glu 195 200 205 His Gly Ala Gln His His Val Tyr Lys Leu Val Lys Asp 210 215 220 <210> 8 <211> 221 <212> PRT <213> Homo sapiens <400> 8 Trp Gln Gly Arg Ser Val Ala Ser Ser Lys Leu Trp Met Leu Glu Phe 1 5 10 15 Ser Ala Phe Leu Glu Gln Gln Gln Asp Pro Asp Thr Tyr Asn Lys His 20 25 30 Leu Phe Val His Ile Gly Gln Ser Ser Pro Ser Tyr Ser Asp Pro Tyr 35 40 45 Leu Glu Ala Val Asp Ile Arg Gln Ile Tyr Asp Lys Phe Pro Glu Lys 50 55 60 Lys Gly Gly Leu Lys Asp Leu Phe Glu Arg Gly Pro Ser Asn Ala Phe 65 70 75 80 Phe Leu Val Lys Phe Trp Ala Asp Leu Asn Thr Asn Ile Glu Asp Glu 85 90 95 Gly Ser Ser Phe Tyr Gly Val Ser Ser Gln Tyr Glu Ser Pro Glu Asn 100 105 110 Met Ile Ile Thr Cys Ser Thr Lys Val Cys Ser Phe Gly Lys Gln Val 115 120 125 Val Glu Lys Val Glu Thr Glu Tyr Ala Arg Tyr Glu Asn Gly His Tyr 130 135 140 Ser Tyr Arg Ile His Arg Ser Pro Leu Cys Glu Tyr Met Ile Asn Phe 145 150 155 160 Ile His Lys Leu Lys His Leu Pro Glu Lys Tyr Met Met Asn Ser Val 165 170 175 Leu Glu Asn Phe Thr Ile Leu Gln Val Val Thr Asn Arg Asp Thr Gln 180 185 190 Glu Thr Leu Leu Cys Ile Ala Tyr Val Phe Glu Val Ser Ala Ser Glu 195 200 205 His Gly Ala Gln His His Ile Tyr Arg Leu Val Lys Glu 210 215 220 <210> 9 <211> 6 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <220> <221> MOD_RES <222> (2)..(2) <223> Any amino acid <220> <221> MOD_RES <222> (4)..(5) <223> Any amino acid<400> 9 His Xaa Arg Xaa Xaa Ser 1 5

Claims (53)

A compound of formula (I):

In the above equation,
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 -N(R)-, -O- or -C(O) - is independently and optionally replaced by;
Ring A is
is selected from, each of which is optionally substituted;
Ring B is
is selected from;
Each R ^w is independently is selected from;
Each R ² is independently -OR, -C(O)NR ₂ , optionally substituted -C _1-6 aliphatic, is selected from;
Each Y is independently N or CR ⁵ ;
each R ³ is independently H, or 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 an optionally substituted 5- having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. is a 6-membered heteroaryl;
Each m is independently 0, 1, or 2;
p is 0, 1, or 2;
Each R is independently selected from H, optionally substituted -C _1-6 aliphatic, optionally substituted 3-8 membered saturated or partially unsaturated monocyclic carbocyclyl, or nitrogen, oxygen or sulfur. It is an optionally substituted 3-8 membered saturated or partially unsaturated monocyclic heterocyclyl having 2 heteroatoms. The compound according to claim 1, wherein L ¹ is a C _1-6 divalent straight or branched hydrocarbon chain, and 1, 2 or 3 methylene units of the chain are replaced with -N(R)-, or a pharmaceutically active compound thereof. Salts allowed. The method of claim 1 or 2, wherein ring A is optionally substituted. A phosphorus compound, or a pharmaceutically acceptable salt thereof. The method of claim 1 or 2, wherein ring A is optionally substituted. A phosphorus compound, or a pharmaceutically acceptable salt thereof. The method of claim 1 or 2, wherein ring A is optionally substituted. A phosphorus compound, or a pharmaceutically acceptable salt thereof. The method of claim 1 or 2, wherein ring A is optionally substituted. A phosphorus compound, or a pharmaceutically acceptable salt thereof. The method according to any one of claims 1 to 6, wherein ring B is A phosphorus compound, or a pharmaceutically acceptable salt thereof. The method according to any one of claims 1 to 7, wherein each R ^w is independently A phosphorus compound, or a pharmaceutically acceptable salt thereof. The compound according to claim 1, wherein the compound has the formula:




In the above formula, 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. The compound according to claim 1, wherein the compound has the formula:





In the above formula, 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. The compound according to claim 1, wherein the compound has the formula:






In the above formula, 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. The compound according to claim 1, wherein the compound has the formula:


In the above formula, 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. The compound according to claim 1, wherein the compound has the formula:


or
In the above formula, 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. The compound according to claim 1, wherein the compound has the formula:


In the above formula, 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. The compound according to claim 1, wherein the compound has the formula:

In the above formula, 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. The compound according to any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof, selected from the compounds in Table 1. A pharmaceutical composition comprising the compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle. As a method of treating cancer in a patient,
A method comprising administering the compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 17 to a patient. 19. The method of claim 18, wherein the cancer is associated with increased TEAD expression. The method of claim 18 or 19, wherein the cancer is associated with increased TEAD activity. As a method of inhibiting the progression of cancer in a patient,
A method comprising administering the compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 17 to a patient. 22. The method of claim 21, wherein the cancer is associated with increased TEAD expression. The method of claim 21 or 22, wherein the cancer is associated with increased TEAD activity. A method of treating a patient with a disease or disorder associated with increased TEAD expression, comprising:
A method comprising administering a therapeutically effective amount of the compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 17 to a patient in need thereof. A method of treating a patient with a disease or disorder associated with increased TEAD activity, comprising:
A method comprising administering a therapeutically effective amount of the compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 17 to a patient in need thereof. As a method of treating a disease or disorder in which inhibition of TEAD activity is beneficial, comprising:
A method comprising administering a therapeutically effective amount of the compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 17 to a patient in need thereof. As a method of treating a disease or disorder in which Hippo pathway inhibition is beneficial, comprising:
A method comprising administering a therapeutically effective amount of the compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 17 to a patient in need thereof. The method of any one of claims 24 to 27, wherein the disease or disorder is a cell proliferation disorder. 29. The method of claim 28, wherein the cell proliferation 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 cancer cells. The method of any one of claims 19, 20, and 22-25, wherein the increased TEAD expression or increased TEAD1 activity is increased TEAD1 expression or increased TEAD1 activity. The method of any one of claims 19, 20, and 22-25, wherein the increased TEAD expression or increased TEAD activity is increased TEAD2 expression or increased TEAD2 activity. The method of any one of claims 19, 20, and 22-25, wherein the increased TEAD expression or increased TEAD activity is increased TEAD3 expression or increased TEAD3 activity. The method of any one of claims 19, 20, and 22-25, wherein the increased TEAD expression or increased TEAD activity is increased TEAD4 expression or increased TEAD4 activity. The method of any one of claims 19, 20, and 22-25, wherein the increased TEAD expression or increased TEAD1 activity is increased TEAD1 expression or increased TEAD1 activity; increased TEAD2 expression or increased TEAD2 activity; increased TEAD3 expression or increased TEAD3 activity; increased TEAD4 expression or increased TEAD4 activity; or a combination thereof. Use of the compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 17 for treating cancer in a patient. 37. Use according to claim 36, wherein said cancer is associated with increased TEAD expression. Use according to claims 36 or 37, wherein the cancer is associated with increased TEAD activity. Use of the compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 17, for inhibiting the progression of cancer in a patient. The use of claim 39, wherein the cancer is associated with increased TEAD expression. Use according to claims 39 or 40, wherein the cancer is associated with increased TEAD activity. Use of the compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 17, for treating a disease or disorder associated with increased TEAD expression. Use of the compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 17, for treating a disease or disorder associated with increased TEAD activity. Use of the compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 17, for treating a disease or disorder in which inhibition of TEAD activity is beneficial. Use of the compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 17, for treating a disease or disorder in which Hippo pathway inhibition is beneficial. The use according to any one of claims 42 to 45, wherein the disease or disorder is a cell proliferation disorder. 47. The use of claim 46, wherein the cell proliferation 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. Use according to any one of claims 37, 38 and 40-43, wherein the increased TEAD expression or increased TEAD1 activity is increased TEAD1 expression or increased TEAD1 activity. Use according to any one of claims 37, 38 and 40-43, wherein the increased TEAD expression or increased TEAD activity is increased TEAD2 expression or increased TEAD2 activity. Use according to any one of claims 37, 38 and 40 to 43, wherein the increased TEAD expression or increased TEAD activity is increased TEAD3 expression or increased TEAD3 activity. Use according to any one of claims 37, 38 and 40-43, wherein the increased TEAD expression or increased TEAD activity is increased TEAD4 expression or increased TEAD4 activity. The method of any one of claims 37, 38, and 40-43, wherein the increased TEAD expression or increased TEAD activity comprises increased TEAD1 expression or increased TEAD1 activity; increased TEAD2 expression or increased TEAD2 activity; increased TEAD3 expression or increased TEAD3 activity; increased TEAD4 expression or increased TEAD4 activity; or a combination thereof.

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