KR20230172548A - MEK inhibitors and uses thereof - Google Patents

Abstract

The present invention provides MEK inhibitors of formula (I') , compositions thereof, and methods of using the same:

Description

MEK inhibitors and uses thereof

Cross-reference to related applications

This application claims priority to U.S. Provisional Application No. 63/175,837, filed April 16, 2021, and U.S. Provisional Application No. 63/262,093, filed October 5, 2021, each of which is incorporated herein by reference in its entirety. is incorporated into.

The present invention relates to compounds and methods useful for inhibiting mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK). The invention also provides pharmaceutically acceptable compositions comprising compounds of the invention and methods of using such compositions in the treatment of proliferative disorders, such as cancer.

Activation of the p42/44 MAPK signaling pathway, including mitogen-activated protein kinase/extracellular signal-regulated kinase (ERK) kinase (MEK)-ERK, has been implicated in the development and progression of various cancers. The MEK-ERK pathway is often activated by mutations in the upstream factors BRAF or Ras, or by signaling from constitutively activated cell surface receptors. Inhibition of MEK may be a promising strategy for controlling the growth of tumors, e.g., tumors involving MEK pathway signaling.

It has now been discovered that the compounds of the present invention, and pharmaceutically acceptable compositions thereof, are effective as MEK inhibitors.

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

In the above equation, each variable is as defined and described herein.

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

In the above equation, each variable is as defined and described herein.

Another aspect of the invention provides a method of treating a MEK-mediated disorder in a subject. The methods include administering to a subject in need thereof a therapeutically effective amount of a compound described herein to treat a disorder mediated by MEK, as further described in the detailed description.

Another aspect of the invention provides a method of inhibiting MEK activity. The method includes contacting MEK or KSR-MEK complex or RAF-MEK complex with an effective amount of a compound described herein, as further described in the detailed description. In some embodiments, the methods provided herein include contacting the MEK-KSR-RAF complex with an effective amount of a compound described herein, as further described in the detailed description.

In some aspects, the compounds of the invention, and pharmaceutically acceptable compositions thereof, are useful for treating proliferative disorders, such as cancer, as described herein.

1. General Description of Certain Embodiments of the Invention:

The compounds of the present invention, and pharmaceutical compositions thereof, are useful as inhibitors of MEK. Without wishing to be bound by any particular theory, the compounds of the present invention may directly engage members of the RAF family of proteins at the MEK interface, and RAF family members, particularly BRAF or CRAF, may be MEK inhibitors in the RAF-MEK complex. It is believed that the prototypical and three-dimensionally different (allosteric) pockets can be remodeled. Without wishing to be bound by any particular theory, it is believed that the compounds of the invention are capable of binding to the RAF-MEK complex and disrupting the associated RAF-MEK complex.

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

In the above equation,

Ring A is is selected from;

each independently represents a single bond or a double bond;

each R ¹ is independently H, halogen, -CN, or optionally substituted C _1-6 aliphatic;

X is C, CH, or N;

Y is CR ⁶ , C(O), or N;

L' is a covalent bond, -O-, -C(R ⁹ ) ₂ -, or -NR ⁸ -;

Each of R ² , R ⁵ , and R ⁸ is independently H or an optionally substituted C _1-6 aliphatic;

Each of R ³ , R ⁴ , R ⁶ , and R ⁹ is independently H, halogen, or optionally substituted C _1-6 aliphatic;

Ring B is a 3-8 membered monocyclic carbocyclic ring, a 3-8 membered monocyclic heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S, 5-11 A 5-11 membered bicyclic heterocyclic ring having 1-5 heteroatoms independently selected from a bicyclic heterocyclic ring, N, O, or S, a phenyl ring, N, O, or S a 5-6 membered monocyclic heteroaromatic ring having 1-3 heteroatoms independently selected from, an 8-11 membered bicyclic aromatic ring, and 1-5 members independently selected from N, O, or S. an optionally substituted ring selected from 8-11 membered bicyclic heteroaromatic rings having heteroatoms;

L is a covalent bond, or 1, 2 or 3 methylene units of the chain are independently and optionally -O-, -S-, -N(R)-, -N ⁺ (R) ₂ -, -CH(R )-, -CH(OR)-, -CH(SR)-, -CH(N(R) ₂ )-, -C(=NR)-, -C(=N-OR)-, -C(O )-, -S(O) ₂ -, -(CH ₂ -CH ₂ -O) _1-10 -, -S(O)-, -N(R)-C(O)-, -N(R) -S(O) ₂ -, -C(O)-N(R)-, -S(O) ₂ -N(R)-, -P(O)(OR)-, or -Cy- C _1-10 2 is a straight or branched hydrocarbon chain;

Each -Cy- is independently a 3-7 membered carbocyclic ring, a 3-7 membered heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S, a phenyl ring, and N is an optionally substituted ring selected from a 5-6 membered heteroaromatic ring having 1-3 heteroatoms independently selected from , O, or S;

R ⁷ is R, -CN, -S(O) ₂ -N(R) ₂ , -NR-S(O) ₂ -R, -P(O)(OR)-N(R) ₂ , -C( =NR)-N(R) ₂ , -C(=N-OR)-N(R) ₂ , -C(O)-R, -S(O)-R, -N(R) ₂ , -OR , -SR, or ego;

Each R is independently H, -CN, or optionally substituted C _1-6 aliphatic;

n is 0, 1, 2, 3, 4, or 5.

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

In the above equation,

Ring A is is selected from;

each independently represents a single bond or a double bond;

each R ¹ is independently H, halogen, -CN, or optionally substituted C _1-6 aliphatic;

X is C, CH, or N;

Y is CR ⁶ , C(O), or N;

L' is a covalent bond, -O-, -C(R ⁹ ) ₂ -, or -NR ⁸ -;

Each of R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁸ , and R ⁹ is independently H or an optionally substituted C _1-6 aliphatic;

Ring B is a 3-8 membered monocyclic carbocyclic ring, a 3-8 membered monocyclic heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S, 5-11 A 5-11 membered bicyclic heterocyclic ring having 1-5 heteroatoms independently selected from a bicyclic carbocyclic ring, N, O, or S, a phenyl ring, N, O, or S a 5-6 membered monocyclic heteroaromatic ring having 1-3 heteroatoms independently selected from, an 8-11 membered bicyclic aromatic ring, and 1-5 members independently selected from N, O, or S. an optionally substituted ring selected from 8-11 membered bicyclic heteroaromatic rings having heteroatoms;

L is a covalent bond, or 1, 2 or 3 methylene units of the chain are independently and optionally -O-, -S-, -N(R)-, -N ⁺ (R) ₂ -, -CH(R )-, -CH(OR)-, -CH(SR)-, -CH(N(R) ₂ )-, -C(=NR)-, -C(=N-OR)-, -C(O )-, -S(O) ₂ -, -(CH ₂ -CH ₂ -O) _1-10 -, -S(O)-, -N(R)-C(O)-, -N(R) -S(O) ₂ -, -C(O)-N(R)-, -S(O) ₂ -N(R)-, -P(O)(OR)-, or -Cy- C _1-10 2 is a straight or branched hydrocarbon chain;

Each -Cy- is independently a 3-7 membered carbocyclic ring, a 3-7 membered heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S, a phenyl ring, and N is an optionally substituted ring selected from a 5-6 membered heteroaromatic ring having 1-3 heteroatoms independently selected from , O, or S;

R ⁷ is R, -CN, -S(O) ₂ -N(R) ₂ , -NR-S(O) ₂ -R, -P(O)(OR)-N(R) ₂ , -C( =NR)-N(R) ₂ , -C(=N-OR)-N(R) ₂ , -C(O)-R, -S(O)-R, -N(R) ₂ , -OR , -SR, or ego;

Each R is independently H, -CN, or optionally substituted C _1-6 aliphatic;

n is 0, 1, 2, 3, 4, or 5.

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

In the above equation,

Ring A is is selected from;

each independently represents a single bond or a double bond;

each R ¹ is independently H, halogen, -CN, or optionally substituted C _1-6 aliphatic;

X is C or N;

Y is CR ⁶ , C(O), or N;

Each of R ² , R ³ , R ⁴ , R ⁵ , and R ⁶ is independently H or an optionally substituted C _1-6 aliphatic;

Ring B is a phenyl ring, a 5-6 membered heteroaromatic ring having 1-3 heteroatoms independently selected from N, O, or S, an 8-10 membered bicyclic aromatic ring, and N, O, or an optionally substituted ring selected from 8-10 membered bicyclic heteroaromatic rings having 1-5 heteroatoms independently selected from S;

L is a covalent bond, or 1, 2 or 3 methylene units of the chain are independently and optionally -O-, -S-, -N(R)-, -N ⁺ (R) ₂ -, -CH(R )-, -CH(OR)-, -CH(SR)-, -CH(N(R) ₂ )-, -C(=NR)-, -C(=N-OR)-, -C(O )-, -S(O) ₂ -, -P(O)(OR)-, or -Cy- where C _1-10 2 is a straight or branched hydrocarbon chain;

Each -Cy- is independently a 3-7 membered carbocyclic ring, a 3-7 membered heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S, a phenyl ring, and N is an optionally substituted ring selected from a 5-6 membered heteroaromatic ring having 1-3 heteroatoms independently selected from , O, or S;

R ⁷ is -CN, -S(O) ₂ -N(R) ₂ , -NR-S(O) ₂ -R, -P(O)(OR)-N(R) ₂ , -C(=NR )-N(R) ₂ , -C(=N-OR)-N(R) ₂ , -C(O)-R, -N(R) ₂ , -OR, -SR, or ego;

Each R is independently H, -CN, or optionally substituted C _1-6 aliphatic;

n is 0, 1, 2, 3, 4, or 5.

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 1-6 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-5 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. In another embodiment, the aliphatic group contains 1-3 aliphatic carbon atoms, and in another embodiment, the aliphatic group contains 1-2 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 are optionally substituted, and include nitrogen (including N-oxides), oxygen, sulfur (including oxidized forms such as sulfones and sulfonates), phosphorus (including oxidized forms such as phosphates), It may be selected from boron and the like. 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 basic nitrogen; or a replaceable nitrogen of a heterocyclic ring, 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,” refers to a monocyclic or cyclic group having a total of 5 to 14 ring members. Refers to a bicyclic ring system, where at least one ring in the system is aromatic and each ring in the system contains 3 to 7 ring members. The term “aryl” may be used interchangeably with the term “aryl ring”. In certain embodiments of the 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, are within the scope of the term "aryl" as used herein. Also included within.

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 to 7 membered monocyclic or 7 to 10 membered bicyclic heterocyclic moiety having one or more, preferably 1 to 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, the nitrogen is N (as in 3,4-dihydro-2 H -pyrrolyl), (p NH (as in rolidinyl), 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 and , groups in which the heterocyclyl ring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as indolinyl, 3 H- 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 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°, taken together with their intervening atom(s), form a 3- heteroatom having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Forms a 12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring, which may be substituted by a divalent substituent on the saturated carbon atom at 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 ^ㆍ is a 5-6 membered group having _0-4 heteroatoms independently selected from C _1-4 aliphatic, -CH ₂ Ph, -O(CH ₂ ) _0-1 Ph, or nitrogen, oxygen, or sulfur. is independently selected from a saturated, partially unsaturated, or aryl ring, wherein each R ^· is unsubstituted or, if preceded by halo, substituted only with 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-, where each independently occurring R ^* is hydrogen, C _1-6 aliphatic, or unsubstituted 5-6 membered saturated, partially unsaturated, or unsubstituted having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. is selected from an aryl 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 not substituted or, if preceded by halo, 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; where 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 ^† , taken together with their intervening atom(s), form an unsubstituted ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Forms a 3- to 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 independently selected from a _5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, wherein each R ^ㆍ is not substituted or, if preceded by halo, is substituted only with 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, where appropriate, contain non-toxic ammoniums, quaternary ammoniums, and counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates and aryl sulfonates. Contains amine cations formed using.

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.

As used herein, the term “provided compound” refers to any MEK inhibitor genus, subgenus, and/or species specified herein.

As used herein, the term “inhibitor” or “MEK inhibitor” or “MEK antagonist” is defined as a compound that binds and/or inhibits MEK with measurable affinity. In some embodiments, inhibition in the presence of a MEK inhibitor or MEK antagonist 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. has

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 MEK, and MEK in the absence of the compound, or composition thereof. means a measurable change or inhibition in MEK activity between equivalent samples comprising.

As used herein, the term “effective amount” refers to an amount of a compound sufficient to produce a beneficial or desired result (e.g., a therapeutic, ameliorative, inhibitory, or prophylactic result). The effective amount may be administered in one or more administrations, applications, or dosages, and is not limited to a specific formulation or route of administration.

As used herein, the term “treating” includes alleviating, reducing, controlling, ameliorating or eliminating any effect, e.g., resulting in amelioration of the condition, disease, disorder, etc., or amelioration of the symptoms thereof. . 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 continue after symptoms have resolved, for example, to prevent or delay recurrence.

As used herein, the phrases “MEK-mediated disorder” or “MEK-mediated disease” or “MEK-related disease or disorder” refers to a disease or disorder associated with or mediated by MEK or MEK activity. do. A non-limiting example of a MEK-related disease or disorder is MEK-related cancer.

As used herein, the term “pharmaceutical composition” refers to a combination of an active agent with an inert or active carrier that makes the composition particularly suitable for diagnostic or therapeutic use in vivo or in vitro.

As used herein, the term “pharmaceutically acceptable carrier” refers to standard carriers such as phosphate buffered saline solutions, water, emulsions (e.g., oil/water or water/oil emulsions), and various types of wetting agents. refers to any of the pharmaceutical carriers. The composition may also include stabilizers and preservatives. For examples of carriers, stabilizers and adjuvants, see, e.g., Martin, Remington's Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, PA [1975].

As used herein, a “non-ATP competitive” or “ATP non-competitive” MEK inhibitor refers to an inhibitor of MEK that does not bind to the ATP pocket of MEK or displace ATP from the MEK active site, and the MEK-ATP complex When co-bonded to, they can form direct contact. Non-ATP competitive inhibition by the compounds of the invention can be confirmed by art-recognized methods such as enzymological studies, competition assays, and biophysical methods, including X-ray co-crystallography. Exemplary non-ATP competitive inhibitors of the invention inhibit recombinant MEK1 or MEK2 with an IC ₅₀ of about 1 nM to about 50 μM. In some embodiments, exemplary non-ATP competitive inhibitors of the invention have about 1 nM to about 1000 nM, about 1 μM to about 50 μM, about 1 μM to about 25 μM, about 1 μM to about 10 μM, about 500 μM. Inhibits recombinant MEK1 or MEK2 with an IC ₅₀ of nM to about 5 μM, or about 10 nM to about 500 nM.

As used herein, “inhibitor pocket” refers to a structure formed at the interface of the interaction between MEK and KSR or BRAF or CRAF into which an inhibitor of the invention engages.

As used herein, a compound of the invention "binds sterically differently to the inhibitor pocket" when the compound binds outside the active site, including, for example, a site external to or adjacent to the ATP-binding site of the kinase. do".

As used herein, an "inhibitor-inhibitor pocket complex" means that an inhibitor of the invention binds human MEK (MEK1 or MEK2) adjacent to ATP in a physiological complex between MEK and KSR and a human kinase inhibitor of Ras (KSR1, KSR2, or KSR homologs (BRAF or CRAF) describe species that bind sterically differently to the inhibitor pocket formed at the interaction interface.

As used herein, when a moiety on an inhibitor "engages" an amino acid residue of MEK and/or KSR and/or BRAF or CRAF in the inhibitor pocket, this interaction is determined by X-ray crystallography, or similar It can be detected by structural methods, such as cryo-electron microscopy, NMR, or in silico docking involving fragment binding and computational simulations, which show that the interactions that define the engagement are, for example, about 2 Å to about 5 Å. , or less than or equal to about 8 Å, including from about 5 Å to about 8 Å, demonstrating a separation between the inhibitor moiety and the amino acid residue. The distances given here allow for hydrogen atoms to be implicitly included; However, hydrogen atoms are not included in this crystallographic model, which is appropriate unless the crystal diffracts at very high resolution (i.e. better than 1.5 Angstroms). A literature search of drug-receptor atom pairs across all structures in the Protein Data Bank used a 4-5 Angstrom distance cutoff (PMID 29308120, 26517868, 19221587) to estimate reasonable small molecule hydrophobic binding interactions; Among the most frequently occurring drug-receptor atom pairs, we found that the intermolecular carbon-carbon interaction was similar to the trametinib-KSR contact. Regarding the interaction between the inhibitors of the invention and the MEK-KSR and/or BRAF or CRAF complexes, a 4 Angstrom contact is reasonable based on the nature of the trametinib-KSR interaction and the known priorities of drug-receptor complexes. . This contact is within the range of known contacts as defined by several independent groups (PMID 29308120, 26517868, 19221587).

3. Description of Exemplary Embodiments:

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

In the above equation,

Ring A is is selected from;

each independently represents a single bond or a double bond;

each R ¹ is independently H, halogen, -CN, or optionally substituted C _1-6 aliphatic;

X is C, CH, or N;

Y is CR ⁶ , C(O), or N;

L' is a covalent bond, -O-, -C(R ⁹ ) ₂ -, or -NR ⁸ -;

Each of R ² , R ⁵ , and R ⁸ is independently H or an optionally substituted C _1-6 aliphatic;

Each of R ³ , R ⁴ , R ⁶ , and R ⁹ is independently H, halogen, or optionally substituted C _1-6 aliphatic;

Ring B is a 3-8 membered monocyclic carbocyclic ring, a 3-8 membered monocyclic heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S, 5-11 a 5-11 membered bicyclic carbocyclic ring having 1-5 heteroatoms independently selected from a 1-membered bicyclic carbocyclic ring, N, O, or S, a phenyl ring, N, O, or S a 5-6 membered monocyclic heteroaromatic ring having 1-3 heteroatoms independently selected from, an 8-11 membered bicyclic aromatic ring, and 1-5 members independently selected from N, O, or S. an optionally substituted ring selected from 8-11 membered bicyclic heteroaromatic rings having heteroatoms;

L is a covalent bond, or 1, 2 or 3 methylene units of the chain are independently and optionally -O-, -S-, -N(R)-, -N ⁺ (R) ₂ -, -CH(R )-, -CH(OR)-, -CH(SR)-, -CH(N(R) ₂ )-, -C(=NR)-, -C(=N-OR)-, -C(O )-, -S(O) ₂ -, -(CH ₂ -CH ₂ -O) _1-10 -, -S(O)-, -N(R)-C(O)-, -N(R) -S(O) ₂ -, -C(O)-N(R)-, -S(O) ₂ -N(R)-, -P(O)(OR)-, or -Cy- C _1-10 2 is a straight or branched hydrocarbon chain;

Each -Cy- is independently a 3-7 membered carbocyclic ring, a 3-7 membered heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S, a phenyl ring, and N is an optionally substituted ring selected from a 5-6 membered heteroaromatic ring having 1-3 heteroatoms independently selected from , O, or S;

R ⁷ is R, -CN, -S(O) ₂ -N(R) ₂ , -NR-S(O) ₂ -R, -P(O)(OR)-N(R) ₂ , -C( =NR)-N(R) ₂ , -C(=N-OR)-N(R) ₂ , -C(O)-R, -S(O)-R, -N(R) ₂ , -OR , -SR, or ego;

Each R is independently H, -CN, or optionally substituted C _1-6 aliphatic;

n is 0, 1, 2, 3, 4, or 5.

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

In the above equation,

Ring A is is selected from;

each independently represents a single bond or a double bond;

each R ¹ is independently H, halogen, -CN, or optionally substituted C _1-6 aliphatic;

X is C, CH, or N;

Y is CR ⁶ , C(O), or N;

Each of R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , and R ⁸ is independently H or an optionally substituted C _1-6 aliphatic;

Ring B is a 3-8 membered monocyclic carbocyclic ring, a 3-8 membered monocyclic heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S, 5-11 a 5-11 membered bicyclic carbocyclic ring having 1-5 heteroatoms independently selected from a 1-membered bicyclic carbocyclic ring, N, O, or S, a phenyl ring, N, O, or S a 5-6 membered monocyclic heteroaromatic ring having 1-3 heteroatoms independently selected from, an 8-11 membered bicyclic aromatic ring, and 1-5 members independently selected from N, O, or S. an optionally substituted ring selected from 8-11 membered bicyclic heteroaromatic rings having heteroatoms;

L is a covalent bond, or 1, 2 or 3 methylene units of the chain are independently and optionally -O-, -S-, -N(R)-, -N ⁺ (R) ₂ -, -CH(R )-, -CH(OR)-, -CH(SR)-, -CH(N(R) ₂ )-, -C(=NR)-, -C(=N-OR)-, -C(O )-, -S(O) ₂ -, -(CH ₂ -CH ₂ -O) _1-10 -, -S(O)-, -N(R)-C(O)-, -N(R) -S(O) ₂ -, -C(O)-N(R)-, -S(O) ₂ -N(R)-, -P(O)(OR)-, or -Cy- C _1-10 2 is a straight or branched hydrocarbon chain;

Each -Cy- is independently a 3-7 membered carbocyclic ring, a 3-7 membered heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S, a phenyl ring, and N is an optionally substituted ring selected from a 5-6 membered heteroaromatic ring having 1-3 heteroatoms independently selected from , O, or S;

R ⁷ is R, -CN, -S(O) ₂ -N(R) ₂ , -NR-S(O) ₂ -R, -P(O)(OR)-N(R) ₂ , -C( =NR)-N(R) ₂ , -C(=N-OR)-N(R) ₂ , -C(O)-R, -S(O)-R, -N(R) ₂ , -OR , -SR, , or a fluorescent probe;

Each R is independently H, -CN, or optionally substituted C _1-6 aliphatic;

n is 0, 1, 2, 3, 4, or 5.

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

In the above equation,

Ring A is is selected from;

each independently represents a single bond or a double bond;

each R ¹ is independently H, halogen, -CN, or optionally substituted C _1-6 aliphatic;

X is C or N;

Y is CR ⁶ , C(O), or N;

Each of R ² , R ³ , R ⁴ , R ⁵ , and R ⁶ is independently H or an optionally substituted C _1-6 aliphatic;

Ring B is a phenyl ring, a 5-6 membered heteroaromatic ring having 1-3 heteroatoms independently selected from N, O, or S, an 8-10 membered bicyclic aromatic ring, and N, O, or an optionally substituted ring selected from 8-10 membered bicyclic heteroaromatic rings having 1-5 heteroatoms independently selected from S;

L is a covalent bond, or 1, 2 or 3 methylene units of the chain are independently and optionally -O-, -S-, -N(R)-, -N ⁺ (R) ₂ -, -CH(R )-, -CH(OR)-, -CH(SR)-, -CH(N(R) ₂ )-, -C(=NR)-, -C(=N-OR)-, -C(O )-, -S(O) ₂ -, -P(O)(OR)-, or -Cy- where C _1-10 2 is a straight or branched hydrocarbon chain;

Each -Cy- is independently a 3-7 membered carbocyclic ring, a 3-7 membered heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S, a phenyl ring, and N is an optionally substituted ring selected from a 5-6 membered heteroaromatic ring having 1-3 heteroatoms independently selected from , O, or S;

R ⁷ is -CN, -S(O) ₂ -N(R) ₂ , -NR-S(O) ₂ -R, -P(O)(OR)-N(R) ₂ , -C(=NR )-N(R) ₂ , -C(=N-OR)-N(R) ₂ , -C(O)-R, -N(R) ₂ , -OR, -SR, or ego;

Each R is independently H, -CN, or optionally substituted C _1-6 aliphatic;

n is 0, 1, 2, 3, 4, or 5.

As generally defined above, Ring A is wherein each of R ³ and R ⁴ is independently as defined and described in the embodiments herein.

In some embodiments, Ring A is wherein each of R ³ and R ⁴ is independently as defined and described in the embodiments herein.

In some embodiments, Ring A is wherein each of R ³ and R ⁴ is independently as defined and described in the embodiments herein.

In some embodiments, Ring A is and wherein each of R ³ and R ⁴ is independently as defined and described in the embodiments herein. In some embodiments, Ring A is am.

In some embodiments, Ring A is and wherein each of R ³ and R ⁴ is independently as defined and described in the embodiments herein. In some embodiments, Ring A is am.

In some embodiments, Ring A is and where R ³ is as defined and described in the embodiments herein. In some embodiments, Ring A is am.

In some embodiments, Ring A is am.

In some embodiments, Ring A is and wherein each of R ³ and R ⁴ is as defined and described in the embodiments herein. In some embodiments, Ring A is am.

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

As generally defined above, each independently represents a single bond or a double bond.

In some embodiments, represents a single bond. In some embodiments, represents a double bond.

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

As generally defined above, R ¹ is H, halogen, -CN, or optionally substituted C _1-6 aliphatic.

In some embodiments, R ¹ is H.

In some embodiments, R ¹ is halogen. In some embodiments, R ¹ is F. In some embodiments, R ¹ is Cl. In some embodiments, R ¹ is Br. In some embodiments, R ¹ is I.

In some embodiments, R ¹ is -CN.

In some embodiments, R ¹ is optionally C _1-6 aliphatic. In some embodiments, R ¹ is optionally substituted C _1-6 alkyl. In some embodiments, R ¹ is C _1-6 aliphatic substituted 1-6 times with halogen. In some embodiments, R ¹ is C _1-6 alkyl substituted 1-6 times with halogen. In some embodiments, R ¹ is methyl, ethyl, propyl, or isopropyl. In some embodiments, R ¹ is cyclopropyl. In some embodiments, R ¹ is -CH=CH. In some embodiments, R ¹ is am. In some embodiments, R ¹ is am. In some embodiments, R ¹ is -CH ₂ F, -CHF ₂ , or -CF ₃ .

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

As generally defined above, X is C or N.

In some embodiments, X is C. In some embodiments, X is CH. In some embodiments, X is N.

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

As generally defined above, Y is CR ⁶ , C(O), or N.

In some embodiments, Y is CR ⁶ , where R ⁶ is as defined and described in the embodiments herein. In some embodiments, Y is C(O). In some embodiments, Y is N.

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

As generally defined above, L' is a covalent bond, -O-, -C(R ⁹ ) ₂ -, or -NR ⁸ -, where R ⁸ is as defined and described in the embodiments herein. .

In some embodiments, L' is a covalent bond. In some embodiments, L' is -O-. In some embodiments, L' is -C(R ⁹ ) ₂ -. In some embodiments, L' is -CH ₂ -. In some embodiments, L' is -NR ⁸ -. In some embodiments, L' is -NH-.

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

As generally defined above, each of R ² , R ³ , R ⁴ , R ⁵ , and R ⁶ is independently H or an optionally substituted C _1-6 aliphatic.

In some embodiments, R ² is H or optionally C _1-6 aliphatic. In some embodiments, R ² is H. In some embodiments, R ² is optionally substituted C _1-6 aliphatic. In some embodiments, R ² is optionally substituted C _1-6 alkyl. In some embodiments, R ² is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R ² is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R ² is -C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with -OH. In some embodiments, R ² is -C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with -OH. In some embodiments, R ² is methyl, ethyl, propyl, or isopropyl. In some embodiments, R ² is cyclopropyl. In some embodiments, R ² is -CH=CH ₂ . In some embodiments, R ² is am. In some embodiments, R ² is am. In some embodiments, R ² is -CH ₂ F, -CHF ₂ , or -CF ₃ .

In some embodiments, R ³ is H, halogen, or optionally substituted C _1-6 aliphatic. In some embodiments, R ³ is H or optionally substituted C _1-6 aliphatic. In some embodiments, R ³ is H. In some embodiments, R ³ is halogen. In some embodiments, R ³ is -F. In some embodiments, R ³ is -Cl. In some embodiments, R ³ is -I. In some embodiments, R ³ is optionally substituted C _1-6 aliphatic. In some embodiments, R ³ is optionally substituted C _1-6 alkyl. In some embodiments, R ³ is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R ³ is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R ³ is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with —OH. In some embodiments, R ³ is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with —OH. In some embodiments, R ³ is methyl, ethyl, propyl, or isopropyl. In some embodiments, R ³ is cyclopropyl. In some embodiments, R ³ is -CH=CH ₂ . In some embodiments, R ³ is am. In some embodiments, R ³ is am. In some embodiments, R ³ is -CH ₂ F, -CHF ₂ , or -CF ₃ .

In some embodiments, R ⁴ is H, halogen, or optionally substituted C _1-6 aliphatic. In some embodiments, R ⁴ is H or optionally substituted C _1-6 aliphatic. In some embodiments, R ⁴ is H. In some embodiments, R ⁴ is halogen. In some embodiments, R ⁴ is -F. In some embodiments, R ⁴ is -Cl. In some embodiments, R ⁴ is -I. In some embodiments, R ⁴ is optionally substituted C _1-6 aliphatic. In some embodiments, R ⁴ is optionally substituted C _1-6 alkyl. In some embodiments, R ⁴ is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R ⁴ is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R ⁴ is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with —OH. In some embodiments, R ⁴ is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with —OH. In some embodiments, R ⁴ is methyl, ethyl, propyl, or isopropyl. In some embodiments, R ⁴ is cyclopropyl. In some embodiments, R ⁴ is -CH=CH ₂ . In some embodiments, R ⁴ is am. In some embodiments, R ⁴ is am. In some embodiments, R ⁴ is -CH ₂ F, -CHF ₂ , or -CF ₃ .

In some embodiments, R ⁵ is H or optionally substituted C _1-6 aliphatic. In some embodiments, R ⁵ is H. In some embodiments, R ⁵ is optionally substituted C _1-6 aliphatic. In some embodiments, R ⁵ is an optionally substituted 3, 4, 5, or 6 membered saturated or unsaturated carbocyclyl. In some embodiments, R ⁵ is optionally substituted C _1-6 alkyl. In some embodiments, R ⁵ is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R ⁵ is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R ⁵ is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with —OH. In some embodiments, R ⁵ is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with —OH. In some embodiments, R ⁵ is methyl, ethyl, propyl, or isopropyl. In some embodiments, R ⁵ is cyclopropyl. In some embodiments, R ⁵ is -CH=CH ₂ . In some embodiments, R ⁵ is am. In some embodiments, R ⁵ is am. In some embodiments, R ⁵ is -CH ₂ F, -CHF ₂ , or -CF ₃ . In some embodiments, R ⁵ is am. In some embodiments, R ⁵ is am.

In some embodiments, R ⁶ is H, halogen, or optionally substituted C _1-6 aliphatic. In some embodiments, R ⁶ is H or optionally substituted C _1-6 aliphatic. In some embodiments, R ⁶ is H. In some embodiments, R ⁶ is halogen. In some embodiments, R ⁶ is -F. In some embodiments, R ⁶ is -Cl. In some embodiments, R ⁶ is -I. In some embodiments, R ⁶ is optionally substituted C _1-6 aliphatic. In some embodiments, R ⁶ is optionally substituted C _1-6 alkyl. In some embodiments, R ⁶ is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R ⁶ is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R ⁶ is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with —OH. In some embodiments, R ⁶ is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with —OH. In some embodiments, R ⁶ is methyl, ethyl, propyl, or isopropyl. In some embodiments, R ⁶ is cyclopropyl. In some embodiments, R ⁶ is -CH=CH ₂ . In some embodiments, R ⁶ is am. In some embodiments, R ⁶ is am. In some embodiments, R ⁶ is -CH ₂ F, -CHF ₂ , or -CF ₃ .

In some embodiments, R ⁸ is H or optionally substituted C _1-6 aliphatic. In some embodiments, R ⁸ is H. In some embodiments, R ⁸ is optionally substituted C _1-6 aliphatic. In some embodiments, R ⁸ is optionally substituted C _1-6 alkyl. In some embodiments, R ⁸ is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R ⁸ is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R ⁸ is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with —OH. In some embodiments, R ⁸ is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with —OH. In some embodiments, R ⁸ is methyl, ethyl, propyl, or isopropyl. In some embodiments, R ⁸ is cyclopropyl. In some embodiments, R ⁸ is -CH=CH ₂ . In some embodiments, R ⁸ is am. In some embodiments, R ⁸ is am. In some embodiments, R ⁸ is -CH ₂ F, -CHF ₂ , or -CF ₃ . In some embodiments, R ⁸ is selected from those shown in Table 1 below.

In some embodiments, R ⁹ is H, halogen, or optionally substituted C _1-6 aliphatic. In some embodiments, R ⁹ is H or optionally substituted C _1-6 aliphatic. In some embodiments, R ⁹ is H. In some embodiments, R ⁹ is halogen. In some embodiments, R ⁹ is -F. In some embodiments, R ⁹ is -Cl. In some embodiments, R ⁹ is -I. In some embodiments, R ⁹ is optionally substituted C _1-6 aliphatic. In some embodiments, R ⁹ is optionally substituted C _1-6 alkyl. In some embodiments, R ⁹ is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R ⁹ is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R ⁹ is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with —OH. In some embodiments, R ⁹ is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with —OH. In some embodiments, R ⁹ is methyl, ethyl, propyl, or isopropyl. In some embodiments, R ⁹ is cyclopropyl. In some embodiments, R ⁹ is -CH=CH ₂ . In some embodiments, R ⁹ is am. In some embodiments, R ⁹ is am. In some embodiments, R ⁹ is -CH ₂ F, -CHF ₂ , or -CF ₃ . In some embodiments, R ⁹ is selected from those shown in Table 1 below.

In some embodiments, each of R ² , R ³ , R ⁴ , R ⁵ , and R ⁶ is independently selected from those shown in Table 1 below.

As generally defined above, Ring B is a phenyl ring, a 5-6 membered heteroaromatic ring having 1-3 heteroatoms independently selected from N, O, or S, an 8-10 membered bicyclic aromatic ring. rings, and 8-10 membered bicyclic heteroaromatic rings having 1-5 heteroatoms independently selected from N, O, or S.

In some embodiments, Ring B is a 3-8 membered monocyclic carbocyclic ring, a 3-8 membered monocyclic heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S. , a 5-11 membered bicyclic carbocyclic ring, a 5-11 membered bicyclic heterocyclic ring having 1-5 heteroatoms independently selected from N, O, or S, a phenyl ring, N, a 5-6 membered monocyclic heteroaromatic ring having 1-3 heteroatoms independently selected from O, or S, a 8-11 membered bicyclic aromatic ring, and a ring independently selected from N, O, or S It is an optionally substituted ring selected from 8-11 membered bicyclic heteroaromatic rings having 1-5 heteroatoms.

In some embodiments, Ring B is an optionally substituted phenyl ring.

In some embodiments, Ring B is an optionally substituted 5-6 membered heteroaromatic ring having 1-3 heteroatoms independently selected from N, O, or S. In some embodiments, Ring B is an optionally substituted 5-membered heteroaromatic ring having 1, 2, or 3 heteroatoms independently selected from N, O, or S. In some embodiments, Ring B is and It is an optionally substituted 5-membered heteroaromatic ring selected from. In some embodiments, Ring B is substituted with a methyl group. is selected from In some embodiments, Ring B is It is an optionally substituted 5-membered heteroaromatic ring selected from. In some embodiments, Ring B is an optionally substituted 6-membered heteroaromatic ring having 1, 2, or 3 heteroatoms independently selected from N, O, or S. In some embodiments, Ring B is and It is an optionally substituted 6-membered heteroaromatic ring selected from.

In some embodiments, Ring B is an optionally substituted 8-, 9-, or 10-membered bicyclic aromatic ring. In some embodiments, Ring B is an optionally substituted 11-membered bicyclic aromatic ring.

In some embodiments, Ring B is an optionally substituted 8, 9, or 10 membered bicyclic heteroaromatic ring having 1, 2, 3, 4, or 5 heteroatoms independently selected from N, O, or S. . In some embodiments, Ring B is an optionally substituted 11-membered bicyclic heteroaromatic ring having 1, 2, 3, 4, or 5 heteroatoms independently selected from N, O, or S. In some embodiments, Ring B is an optionally substituted 9-membered bicyclic heteroaromatic ring having 1, 2, or 3 heteroatoms independently selected from N, O, or S. In some embodiments, Ring B is an optionally substituted 11-membered bicyclic heteroaromatic ring having 1, 2, or 3 heteroatoms independently selected from N, O, or S. In some embodiments, Ring B is

It is an optionally substituted 9-membered bicyclic heteroaromatic ring selected from. In some embodiments, Ring B is It is an optionally substituted 9-membered bicyclic heteroaromatic ring selected from. In some embodiments, Ring B is It is an optionally substituted 9-membered bicyclic heteroaromatic ring selected from. In some embodiments, Ring B is It is an optionally substituted 9-membered bicyclic heteroaromatic ring selected from. In some embodiments, Ring B is It is an optionally substituted 11-membered bicyclic heteroaromatic ring selected from.

In some embodiments, Ring B is an optionally substituted 3, 4, 5, 6, 7, or 8 membered monocyclic carbocyclic ring. In some embodiments, Ring B is am.

In some embodiments, Ring B is an optionally substituted 3, 4, 5, 6, 7 or 8 membered monocyclic heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S. am. In some embodiments, Ring B is am.

In some embodiments, Ring B is an optionally substituted 5, 6, 7, 8, 9, 10 or 11 membered bicyclic carbocyclic ring. In some embodiments, Ring B is am.

In some embodiments, Ring B is an optionally substituted 5, 6, 7, 8, 9, 10, or 11 membered bicyclic heterocycle having 1-5 heteroatoms independently selected from N, O, or S. It's a click ring. In some embodiments, Ring B is am.

In some embodiments, Ring B is substituted 1-2 times with halogen. In some embodiments, ring B is substituted 1-2 times with R, wherein each R is independently as defined and described in the embodiments herein. In some embodiments, R is optionally substituted C _1-6 alkyl. In some embodiments, R is C _1-6 aliphatic substituted 1-6 times with halogen. In some embodiments, R is C _1-6 alkyl substituted 1-6 times with halogen. In some embodiments, R is methyl, ethyl, propyl, or isopropyl. In some embodiments, R is cyclopropyl. In some embodiments, R is -CH=CH ₂ . In some embodiments, R is am. In some embodiments, R is am. In some embodiments, R is -CH ₂ F, -CHF ₂ , or -CF ₃ .

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

As generally defined above, L is a covalent bond, or 1, 2 or 3 methylene units of the chain are independently and optionally -O-, -S-, -N(R)-, -N ⁺ (R ) ₂ -, -CH(R)-, -CH(OR)-, -CH(SR)-, -CH(N(R) ₂ )-, -C(=NR)-, -C(=N- C _1-10 2 replaced by OR)-, -C(O)-, -S(O) ₂ -, -P(O)(OR)-, or -Cy- is a straight or branched hydrocarbon chain; wherein each of R and -Cy- is independently as defined and described in the embodiments herein.

In some embodiments, L is a covalent bond, or 1, 2, or 3 methylene units of the chain are independently and optionally -O-, -S-, -N(R)-, -N ⁺ (R) ₂ - , -CH(R)-, -CH(OR)-, -CH(SR)-, -CH(N(R) ₂ )-, -C(=NR)-, -C(=N-OR)- , -C(O)-, -S(O) ₂ -, -(CH ₂ -CH ₂ -O) _1-10 -, -S(O)-, -N(R)-C(O)-, -N(R)-S(O) ₂ -, -C(O)-N(R)-, -S(O) ₂ -N(R)-, -P(O)(OR)-, or - C _1-10 2 substituted by Cy- is a straight or branched hydrocarbon chain.

In some embodiments, L is a covalent bond.

In some embodiments, L is a chain wherein 1, 2 or 3 methylene units are independently and optionally -O-, -S-, -N(R)-, -N ⁺ (R) ₂ -, -CH( R)-, -CH(OR)-, -CH(SR)-, -CH(N(R) ₂ )-, -C(=NR)-, -C(=N-OR)-, -C( C _1-10 2 substituted by O)-, -S(O) ₂ -, -P(O)(OR)-, or -Cy- is a straight or branched hydrocarbon chain, where R and -Cy- Each is independently as defined and described in the embodiments herein. In some embodiments, L is 1, 2 or 3 methylene units of the chain independently and optionally O-, -N(R)-, -CH(R)-, -C(=NR)-, -C C _1-10 divalent straight or branched replaced by (=N-OR)-, -C(O)-, -S(O) ₂ -, -P(O)(OR)-, or -Cy- a hydrocarbon chain, wherein each of R and -Cy- is independently as defined and described in the embodiments herein. In some embodiments, R is hydrogen. In some embodiments, R is methyl. In some embodiments, R is -CN. In some embodiments, L is a chain wherein 1, 2 or 3 methylene units are independently and optionally -O-, -S-, -N(R)-, -N ⁺ (R) ₂ -, -CH( R)-, -CH(OR)-, -CH(SR)-, -CH(N(R) ₂ )-, -C(=NR)-, -C(=N-OR)-, -C( O)-, -S(O) ₂ -, -(CH ₂ -CH ₂ -O) _1-10 -, -S(O)-, -N(R)-C(O)-, -N(R )-S(O) ₂ -, -C(O)-N(R)-, -S(O) ₂ -N(R)-, -P(O)(OR)-, or -Cy- C _1-10 2 is a straight or branched hydrocarbon chain, wherein each of R and -Cy- is independently as defined and described in the embodiments herein. In some embodiments, L is one, two or three methylene units of the chain independently and optionally -O-, -N(R)-, -CH(R)-, -C(=NR)-, - C(=N-OR)-, -C(O)-, -S(O) ₂ -, -(CH ₂ -CH ₂ -O) _1-10 -, -S(O)-, -N(R )-C(O)-, -N(R)-S(O) ₂ -, -C(O)-N(R)-, -S(O) ₂ -N(R)-, -P(O )(OR)- or C _1-10 2 substituted by -Cy- is a straight or branched hydrocarbon chain, wherein each of R and -Cy- is independently as defined and described in the embodiments herein. Moiety -(CH ₂ -CH ₂ -O) _1-10 - is -CH ₂ -CH ₂ -O-, -(CH ₂ -CH ₂ -O) ₂ -, -(CH ₂ -CH ₂ -O) ₃ -, -(CH ₂ -CH ₂ -O) ₄ -, -(CH ₂ -CH ₂ -O) ₅ -, -(CH ₂ -CH ₂ -O) ₆ -, -(CH ₂ -CH ₂ - O) ₇ -, -(CH ₂ -CH ₂ -O) ₈ -, -(CH ₂ -CH ₂ -O) ₉ -, or -(CH ₂ -CH ₂ -O) ₁₀ -.

In some embodiments, L is a chain wherein 1, 2 or 3 methylene units are independently and optionally -O-, -S-, -N(R)-, -N ⁺ (R) ₂ -, -CH( R)-, -CH(OR)-, -CH(SR)-, -CH(N(R) ₂ )-, -C(=NR)-, -C(=N-OR)-, -C( C _1-6 2 is a straight or branched hydrocarbon chain replaced by O)-, -S(O) ₂ -, -P(O)(OR)-, or -Cy-, where R and -Cy- Each is independently as defined and described in the embodiments herein. In some embodiments, L is one, two or three methylene units of the chain independently and optionally -O-, -N(R)-, -CH(R)-, -C(=NR)-, - C _1-6 divalent substituted by C(=N-OR)-, -C(O)-, -S(O) ₂ -, -P(O)(OR)-, or -Cy- a topographic hydrocarbon chain, wherein each of R and -Cy- is independently as defined and described in the embodiments herein. In some embodiments, R is hydrogen. In some embodiments, R is methyl. In some embodiments, R is -CN. In some embodiments, L is a chain wherein 1, 2 or 3 methylene units are independently and optionally -O-, -S-, -N(R)-, -N ⁺ (R) ₂ -, -CH( R)-, -CH(OR)-, -CH(SR)-, -CH(N(R) ₂ )-, -C(=NR)-, -C(=N-OR)-, -C( O)-, -S(O) ₂ -, -(CH ₂ -CH ₂ -O) _1-10 -, -S(O)-, -N(R)-C(O)-, -N(R )-S(O) ₂ -, -C(O)-N(R)-, -S(O) ₂ -N(R)-, -P(O)(OR)-, or -Cy- is a C _1-6 divalent straight or branched hydrocarbon chain, wherein each of R and -Cy- is independently as defined and described in the embodiments herein. In some embodiments, L is one, two or three methylene units of the chain independently and optionally -O-, -N(R)-, -CH(R)-, -C(=NR)-, - C(=N-OR)-, -C(O)-, -S(O) ₂ -,-(CH ₂ -CH ₂ -O) _1-10 -, -S(O)-, -N(R )-C(O)-, -N(R)-S(O) ₂ -, -C(O)-N(R)-, -S(O) ₂ -N(R)-, -P(O )(OR)-, or C _1-6 divalent straight or branched hydrocarbon chain replaced by -Cy-, wherein each of R and -Cy- is independently as defined and described in the embodiments herein .

In some embodiments, L is -CH ₂ -, -NH-, -S(O) ₂ -, -NH-S(O) ₂ -, -CH ₂ -S(O) ₂ -, -CH ₂ -P (O)(OH)-, -CH(CF ₃ )-, -CH(CF ₃ )-S(O) ₂ -, -NH-C(O)-NH-, -NH-C(O)-, -O-, -OS(O) ₂ -, is selected from In some embodiments, L is -NH-S(O)-, -N(CH ₃ )-, is selected from

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

As generally defined above, each -Cy- is independently a 3-7 membered carbocyclic ring, a 3-7 membered heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S. an optionally substituted ring selected from cyclic rings, phenyl rings, and 5-6 membered heteroaromatic rings having 1-3 heteroatoms independently selected from N, O, or S.

In some embodiments, -Cy- is an optionally substituted 3, 4, 5, 6 or 7 membered carbocyclic ring.

In some embodiments, -Cy- is an optionally substituted 3, 4, 5, 6, or 7 membered heterocyclic ring having 1, 2, or 3 heteroatoms independently selected from N, O, or S. In some embodiments, -Cy- is am. In some embodiments, -Cy- is am.

In some embodiments, -Cy- is an optionally substituted phenyl ring.

In some embodiments, -Cy- is an optionally substituted 5- or 6-membered heteroaromatic ring having 1, 2, or 3 heteroatoms independently selected from N, O, or S. In some embodiments, -Cy- is am.

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

As generally defined above, R ⁷ is -CN, -S(O) ₂ -N(R) ₂ , -NR-S(O) ₂ -R, -P(O)(OR)-N(R ) ₂ , -C(=NR)-N(R) ₂ , -C(=N-OR)-N(R) ₂ , -C(O)-R, -N(R) ₂ , -OR, - SR, or and wherein each R is independently as defined and described in the embodiments herein.

In some embodiments, R ⁷ is R, -CN, -S(O) ₂ -N(R) ₂ , -NR-S(O) ₂ -R, -P(O)(OR)-N(R) ₂ , -C(=NR)-N(R) ₂ , -C(=N-OR)-N(R) ₂ , -C(O)-R, -S(O)-R, -N(R ) ₂ , -OR, -SR, or am.

In some embodiments, R ⁷ is as defined and described in the embodiments herein. In some embodiments, R ⁷ is -CN. In some embodiments, R ⁷ is -S(O) ₂ -N(R) ₂ . In some embodiments, R ⁷ is -S(O) ₂ -NHR. In some embodiments, R ⁷ is -NR-S(O) ₂ -R. In some embodiments, R ⁷ is -NH—S(O) ₂ -R. In some embodiments, R ⁷ is -P(O)(OR)-N(R) ₂ . In some embodiments, R ⁷ is -P(O)(OH)-N(R) ₂ . In some embodiments, R ⁷ is -P(O)(OR)-NHR. In some embodiments, R ⁷ is -P(O)(OH)-NHR. In some embodiments, R ⁷ is -C(=NR)-N(R) ₂ . In some embodiments, R ⁷ is -C(=NR)-NHR. In some embodiments, R ⁷ is -C(=NH)-N(R) ₂ . In some embodiments, R ⁷ is -C(=N-CN)-N(R) ₂ . In some embodiments, R ⁷ is -C(=NH)-NHR. In some embodiments, R ⁷ is -C(=N-CN)-NHR. In some embodiments, R ⁷ is -C(=N-OR)-N(R) ₂ . In some embodiments, R ⁷ is -C(=N-OR)-NHR. In some embodiments, R ⁷ is -C(=N-OH)-NHR. In some embodiments, R ⁷ is -C(O)-R. In some embodiments, R ⁷ is -S(O)-R. In some embodiments, R ⁷ is -N(R) ₂ . In some embodiments, R ⁷ is -NHR. In some embodiments, R ⁷ is -OR. In some embodiments, R ⁷ is -SR. In some embodiments, R ⁷ is am. In some embodiments, R ⁷ is am. In some embodiments, R ⁷ is am. In some embodiments, R is hydrogen. In some embodiments, R is methyl. In some embodiments, R is -CN.

In some embodiments, R ⁷ is -CN, -NH ₂ , -OH, -NHCH ₃ , is selected from In some embodiments, R ⁷ is -S(O)-CH ₃ , -OC(CH ₃ ) ₃ ,

am. In some embodiments, R ⁷ is am. In some embodiments, R ⁷ is am. In some embodiments, R ⁷ is am. In some embodiments, R ⁷ is, e.g. It is a fluorescent probe containing.

In some embodiments, R ⁷ is selected from those shown in Table 1 below.

As generally defined above, each R is independently H, -CN, or an optionally substituted C _1-6 aliphatic.

In some embodiments, R is H. In some embodiments, R is -CN.

In some embodiments, R is optionally substituted C _1-6 aliphatic. In some embodiments, R is optionally substituted C _1-6 alkyl. In some embodiments, R is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with halogen. In some embodiments, R is C _1-6 aliphatic substituted 1, 2, 3, 4, 5, or 6 times with -OH. In some embodiments, R is C _1-6 alkyl substituted 1, 2, 3, 4, 5, or 6 times with -OH. In some embodiments, R is methyl, ethyl, propyl, or isopropyl. In some embodiments, R is cyclopropyl. In some embodiments, R is -CH=CH ₂ . In some embodiments, R is am. In some embodiments, R is am. In some embodiments, R is -CH ₂ F, -CHF ₂ , or -CF ₃ .

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

As generally defined above, n is 0, 1, 2, 3, 4, or 5.

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, n is 4. In some embodiments, n is 5.

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

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

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

In some embodiments, the invention provides compounds of formula (II-a) to (II-o) :

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

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

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

In some embodiments, the invention provides compounds of formula (III-a) to (III-e) :

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

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

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

In some embodiments, the invention provides compounds of formula (IV-a) to (IV-e) :

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

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 invention provides compounds selected from the starting materials, intermediates, and products described in the Examples herein, or salts thereof.

4. General methods of providing this compound:

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 herein, in the Examples.

In the schemes below where certain protecting groups (“PG”), leaving groups (“LG”) or modification conditions are shown, those skilled in the art will recognize that other protecting groups, leaving groups, and modification conditions are also suitable and contemplated. These groups and transformations are described in March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure , MB Smith and J. March, ^5th Edition, John Wiley & Sons, 2001, Comprehensive Organic Transformations , RC Larock, ^2nd Edition, John Wiley & Sons, 1999, and Protecting Groups in Organic Synthesis , TW Greene and PGM Wuts, 3 ^rd edition, John Wiley & Sons, 1999, each of which is incorporated herein by reference in its entirety.

As used herein, the phrase “leaving group” (LG) refers to halogens (e.g., fluoride, chloride, bromide, iodide), sulfonates (e.g., mesylate, tosylate, benzenesulfonate). , brosylate, nosylate, triflate), diazonium, etc., but are not limited to these.

As used herein, the phrase “oxygen protecting group” includes, for example, carbonyl protecting groups, hydroxyl protecting groups, etc. Hydroxyl protecting groups are well known in the art and are described in Protecting Groups in Organic Synthesis , TW Greene and PGM Wuts, 3 ^rd edition, John Wiley & Sons, 1999, and Philip Kocienski, in "Protecting Groups", Georg Thieme Verlag Stuttgart, New York, 1994, the entirety of which is hereby incorporated by reference. Examples of suitable hydroxyl protecting groups include, but are not limited to, esters, allyl ethers, ethers, silyl ethers, alkyl ethers, arylalkyl ethers, and alkoxyalkyl ethers. Examples of such esters include formates, acetates, carbonates, and sulfonates. Specific examples include formate, benzoyl formate, chloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate, 4 ,4-(ethylene dithio)pentanoate, pivaloate (trimethylacetyl), crotonate, 4-methoxy-crotonate, benzoate, p-benzylbenzoate, 2,4,6-trimethylbenzoate ates, carbonates such as methyl, 9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl, 2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl, vinyl, allyl, and p- Contains nitrobenzyl. Examples of such silyl ethers include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triisopropylsilyl, and other trialkylsilyl ethers. Alkyl ethers include methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, allyl, and allyloxycarbonyl ethers or derivatives. Alkoxyalkyl ethers include acetals such as methoxymethyl, methylthiomethyl, (2-methoxyethoxy)methyl, benzyloxymethyl, beta-(trimethylsilyl)ethoxymethyl, and tetrahydropyranyl ether. Examples of arylalkyl ethers are benzyl, p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyano. Includes nobenzyl, and 2- and 4-picolyl.

Amino protecting groups are well known in the art and are described in Protecting Groups in Organic Synthesis , TW Greene and PGM Wuts, 3 ^rd edition, John Wiley & Sons, 1999, and Philip Kocienski, in "Protecting Groups", Georg Thieme Verlag Stuttgart, New York. , 1994, the entirety of which is hereby incorporated by reference. Suitable amino protecting groups include, but are not limited to, aralkylamines, carbamates, cyclic imides, allyl amines, amides, and the like. Examples of such groups include t-butyloxycarbonyl (BOC), ethyloxycarbonyl, methyloxycarbonyl, trichloroethyloxycarbonyl, allyloxycarbonyl (Alloc), benzyloxocarbonyl (CBZ), allyl, Includes phthalimide, benzyl (Bn), fluorenylmethylcarbonyl (Fmoc), formyl, acetyl, chloroacetyl, dichloroacetyl, trichloroacetyl, phenylacetyl, trifluoroacetyl, benzoyl, etc.

Those skilled in the art will recognize that the various functionalities present in the compounds of the present invention, such as aliphatic groups, alcohols, carboxylic acids, esters, amides, aldehydes, halogens and nitriles, can be reduced, oxidized, esterified, hydrolyzed, partially oxidized, partially reduced, halogenated, dehydrated, etc. It will be appreciated that interconversion may be achieved by techniques well known in the art, including but not limited to partial hydration, and hydration. See, e.g., "March's Advanced Organic Chemistry", ^5th Ed., Ed.: Smith, MB and March, J., John Wiley & Sons, New York: 2001, the entire text of which is incorporated herein by reference. do. Such interconversion may require one or more of the techniques mentioned above, and specific methods for synthesizing the compounds of the invention are described below.

In one aspect, certain compounds of the invention of Formula I , or salts thereof, are prepared generally according to Scheme 1 set forth below:

Scheme 1.

In Scheme 1 above, LG is a leaving group and each variable, e.g. R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁷ , L, n, and Ring B, alone or in combination As defined above and described in the embodiments herein.

As generally shown in Scheme 1 , starting material compound A is converted to compound B , for example, by reaction with triphosgene and R ⁵ -NH ₂ . Then, compound B is e.g. - is converted to compound C by condensation with acetic anhydride to form a ring. Compound C is, for example, POCl ₃ and R ³ NH ₂ and reduced to compound D. Compound D is e.g. After reacting with, for example, the produced alcohol can be converted to compound E by reacting with TsCl and a base to activate the produced alcohol to form a leaving group. Then, compound E is e.g. It is converted to compound F through a coupling reaction with . Compound F can, for example, undergo rearrangement under basic conditions to provide compound G. Compound G is then converted to compound H , for example, by reaction with LR ⁷ .

In some embodiments, the present invention provides synthetic methods as shown in Scheme 1 above. In some embodiments, the invention provides a method of preparing Compound B from Compound A as shown in Scheme 1 above. In some embodiments, the invention provides a method of preparing Compound C from Compound B as shown in Scheme 1 above. In some embodiments, the invention provides a method of preparing Compound D from Compound C as shown in Scheme 1 above. In some embodiments, the invention provides a method of preparing Compound E from Compound D as shown in Scheme 1 above. In some embodiments, the invention provides a method of preparing Compound F from Compound E as shown in Scheme 1 above. In some embodiments, the invention provides a method of preparing Compound G from Compound F as shown in Scheme 1 above. In some embodiments, the invention provides a method of preparing Compound G as shown in Scheme 1 above. A method for preparing compound H is provided.

In some embodiments, the invention provides compounds selected from Compound A , Compound B , Compound C , Compound D , Compound E , Compound F , Compound G , and Compound H , wherein each variable, alone or in combination, is defined above. and is as described in the embodiments herein.

5. 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 MEK, 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 MEK, 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 terms “subject” and “patient” are used interchangeably and refer to the organism to be treated by the methods of the invention. Such organisms preferably include, but are not limited to, mammals (e.g., murine, ape, horse, cow, pig, dog, cat, etc.), and most preferably human.

The term “pharmaceutically acceptable carrier, adjuvant, or vehicle” refers to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound 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. , partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulosic substances, These include, but are not limited to, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and wool fat.

“Pharmaceutically acceptable derivative” means any non-toxic salt, ester, or means a salt or other derivative of an ester.

As used herein, the term “active metabolite or residue thereof” means that the metabolite or residue thereof also inhibits MEK, 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 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 may 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 in 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 can 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 field of pharmaceutical formulations and 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 patients receiving these compositions.

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 judgment of the treating physician and the patient being treated. It should be understood that this will depend on a variety of factors, including the severity of the particular disease. 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

In some embodiments, the invention provides methods of using a compound as described herein to treat a disease or disorder associated with MEK. In some embodiments, the disease or disorder associated with MEK is a proliferative disorder. In some embodiments, the disease or disorder associated with MEK is cancer. In some embodiments, the disease or disorder associated with MEK is cancer, as described herein.

In some aspects and embodiments, characterized by increased MEK expression and/or increased MEK activity, comprising administering a therapeutically effective amount of a compound of the invention or a pharmaceutically acceptable composition thereof to a patient in need thereof. Provided herein are methods of treating a disease or disorder associated with or related thereto, or reducing the severity of the disease or disorder, delaying the onset of the disease or disorder, or inhibiting the progression of the disease or disorder, or one or more symptoms thereof. In some aspects and embodiments, the disease or disorder in which inhibition or antagonism of MEK activity would be beneficial comprises administering a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable composition thereof, to a patient in need thereof. Provided herein are methods of treating, or reducing the severity of, the disease or disorder, delaying the onset of the disease or disorder, or inhibiting the progression of the disease or disorder, or one or more symptoms thereof.

In some embodiments, a compound as described herein is an “ATP non-competitive MEK inhibitor” that stabilizes or “glues” the complex formed between MEK and KSR, and/or BRAF/CRAF. In some embodiments, a compound as described herein comprises a human kinase inhibitor of Ras (KSR1 or KSR2 or the KSR homolog BRAF or CRAF) adjacent to ATP in a physiological complex between MEK and KSR (or BRAF or CRAF) and human MEK. They bind differently to the “inhibitor pocket” formed at the interaction interface between (MEK1 or MEK2), forming an inhibitor-repressor pocket complex. In some embodiments, compounds as described herein are ATP non-competitive kinase inhibitors. In some embodiments, a compound as described herein has a structure such that when bound to an inhibitor-inhibitor pocket complex, the complex comprises the following structural elements: (a) A825 of hKSR1, or P878 of hKSR2, or at least one moiety of the inhibitor that engages R662 of BRAF, or R554 of CRAF, (b) at least one moiety that engages R234 of hMEK1 or R238 of hMEK2, wherein R234 is of hKSR1 or hKSR2 or BRAF or CRAF A moiety within about 5 Å to about 8 Å from any atom. The structures of the complexes described herein, including, for example, the MEK and KSR (or BRAF or CRAF) complexes, and the inhibitor-inhibitor pocket complex, can be found in WO 2021142345, the contents of which are incorporated herein by reference in their entirety. Incorporated by reference. Reference Uniprot sequences for human MEK1, human MEK2, human KSR1, human KSR2, human BRAF, and human CRAF are Uniprot ID Q02750, Uniprot ID P36507, Uniprot ID Q8IVT5, Uniprot ID Q6VAB6, Uniprot ID P15056, and Uniprot ID P04049, respectively. .

In some embodiments, a compound as described herein does not engage one or more of I216 of hMEK1 or I220 of hMEK2 and A825 of KSR1 or P878 of KSR2. In some embodiments, the compound as described herein is an H-bond acceptor, especially an oxygen or nitrogen atom, or a fluorine atom attached to an aromatic ring, or an H bond donor in (a) as described in paragraph above. Includes structural elements according to In some embodiments, the compound as described herein is a moiety of a linker that engages the backbone of A825 of hKSR1, or P878 of hKSR2, or R662 of hBRAF, or R554 of CRAF, directly or through water-mediated contact. Contains structural elements according to (a) as described in the paragraph above.

In some embodiments, compounds as described herein include one or more of the following:

(c) at least one moiety that engages M230 of hMEK1 or M234 of hMEK2, wherein M230 or M234 is the terminal atom of A825 of KSR1 (CB) or (CG) of P878 of hKSR2 or (CG) N661 of hBRAF or a moiety within about 5 Å to about 8 Å of N553 of CRAF;

(d) at least one moiety is an H-bond acceptor or donor that engages directly with the backbone carbonyl of N823 of hKSR1, or T876 of hKSR2, or the backbone amino group of R662 of hBRAF or R554 of hCRAF through water-mediated contact;

(e) at least one moiety that engages Q824 in hKSR1 or Q877 in hKSR2 or Q664 in hBRAF or Q556 in hCRAF;

(f) at least one moiety engaging the side chain atom of A826 in hKSR1 or A879 in hKSR2 or R662 in BRAF or R554 in CRAF;

(g) at least one moiety is a heteroaryl group that engages M143 of hMEK1 or M147 of hMEK2;

(h) at least one moiety is a heteroaryl group that engages F209 of hMEK1 or F213 of hMEK2;

(i) at least one moiety (particularly an H-bond acceptor) engages the backbone amino group of S212 of hMEK1 or S216 of hMEK2;

(j) at least one moiety that engages L215 of hMEK1 or L219 of hMEK2;

(k) at least one moiety that engages I216 of hMEK1 or I220 of hMEK2; and

(l) At least one moiety that engages M219 of hMEK1 or M223 of hMEK2, wherein hMEK1 residues 215-219 adopt a helical structure.

In some embodiments, the moiety corresponding to (c) as described above is selected from substituted or unsubstituted alkyl or cycloalkyl.

In some embodiments, the backbone CO moiety of a compound as described herein engages T876 of hKSR2 or N823 of hKSR1.

In some embodiments, the compounds as described herein engage the binding pocket lined by hMEK1 residues R234 and M230, or hMEK2 residues R238 and M234, and P877 of KSR2 or A825 of KSR1 or R662 of BRAF or R554 of CRAF. all.

In some embodiments, a compound as described herein is a water-mediated H-bond to Arg189 and Arg234 of hMEK1 or to ARG193 and A238 of hMEK2, as well as a pre-helix αG loop -NH of Arg662 of BRAF or ARG 554 of CRAF. It engages the binding pocket through multiple hydrogen bond contacts, including direct H-bonds to the backbone of -.

In some embodiments, a compound as described herein engages A825 of hKSR1 or P878 of hKSR2 or R662 of BRAF or R554 of CRAF. In some embodiments, the compounds as described herein have a distance of from about 5 Å to about 8 Å or less from at least one moiety selected from A825 of hKSR1, P878 of hKSR2, and R662 of BRAF and R554 of CRAF.

Accordingly, in some aspects and embodiments, the invention encompasses administering a MEK inhibitor compound, or pharmaceutical salt or composition thereof, as described herein to a patient in need thereof, including but not limited to a cell proliferative disorder. Provided are methods of treating one or more disorders, diseases, and/or conditions, including but not limited to: In some embodiments, the cell proliferative disorder is cancer. In some embodiments, the cancer is characterized by increased MEK expression and/or increased MEK activity, i.e., “increased activated MEK.”

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 MEK in a sample or cancer or patient refers to a disease or disorder (as determined by techniques known in the art) For example, about 5%, about 10%, about 15%, about 20%, relative to the amount of a substance such as MEK in a control sample or control samples or an internal control, such as an individual or group of individuals not suffering from cancer) 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 20 times, about 25 times, about 50 times, about 100 times or more refers to an increase in the amount of a substance such as MEK. Expression and/or activity of MEK is 1 standard deviation, 2 standard deviations compared to the mean (average) or median value of MEK 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 MEK if it increases by 3 standard deviations, 4 standard deviations, 5 standard deviations or more. As practiced in the art, such control or baseline expression levels may be predetermined or measured prior to measurement in the sample or cancer or subject, or may be obtained from a database of such control samples.

cancer

In some embodiments, the invention provides a method of treating, or preventing or reducing the risk of, cancer in a patient, comprising administering to the patient a compound of the invention, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. do.

As used herein, “cancer” refers to a broad group of various diseases characterized by the uncontrolled growth of abnormal cells in the body. Uncontrolled cell division and growth can lead to the formation of malignant tumors, which can invade surrounding tissues and spread to distant parts of the body through the lymphatic system or bloodstream.

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. In some embodiments, the lung cancer is selected from the group consisting of non-small cell lung cancer, small cell lung cancer, and lung carcinoid tumor.

In some embodiments, the cancer is a K-Ras mutant cancer. In some embodiments, the K-Ras mutant cancer is a K-Ras G12 mutant cancer. In some embodiments, the K-Ras mutant G12 cancer is a K-Ras G12D mutant cancer, a K-Ras G12V mutant cancer, a K-Ras G12C mutant cancer, a K-Ras G12R mutant cancer, or a K-Ras G12S mutant cancer. In some embodiments, the K-Ras mutant cancer is a K-Ras G12D mutant cancer. In some embodiments, the K-Ras mutant cancer is a K-Ras G12V mutant cancer. In some embodiments, the K-Ras mutant cancer is a K-Ras G12C mutant cancer. In some embodiments, the cancer is a mutant B-Raf cancer, such as a B-Raf V600E mutant cancer. In some embodiments, the K-Ras mutant cancer is a K-Ras G13 mutant cancer. In some embodiments, the K-Ras mutant cancer is a K-Ras G13D mutant cancer. In some embodiments, the K-Ras mutant cancer is a K-Ras G12R mutant cancer. In some embodiments, the K-Ras mutant cancer is a K-Ras G12S mutant cancer.

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 mesothelioma, such as malignant mesothelioma. In some embodiments, the cancer is a leukemia (e.g., without limitation, acute leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, acute myeloid leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia), polycythemia vera, lymphoma (e.g., Hodgkin's disease or non-Hodgkin's disease), Waldenstrom's macroglobulinemia, multiple myeloma, heavy chain disease, and sarcomas and carcinomas. Solid tumors (e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendothelioma, synovium, mesothelioma, Ewing tumor, leiomyosarcoma, rhabdomyosarcoma , colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous carcinoma, papillary carcinoma, papillary adenocarcinoma, cystic carcinoma, medullary carcinoma, bronchogenic 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, glioblastoma multiforme ( GBM, also known as glioblastoma), medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma, melanoma, neuroblastoma, and retinoblastoma).

In some embodiments, the cancer is glioma, astrocytoma, glioblastoma multiforme (GBM, also known as glioblastoma), medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, 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, brain stem 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 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 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 related 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 immunodeficiency virus (HIV), which is human T-cell leukemia virus type I (HTLV-I), a highly aggressive form of CD4+ T cell leukemia characterized by clonal integration of HTLV-I in leukemia cells. ) associated solid tumors, human papillomavirus (HPV)-16 positive refractory solid tumors, and virus-related cancers, including adult T-cell leukemia (https://clinicaltrials.gov/ct2/show/study/NCT02631746), as well as 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 methods or uses described herein inhibit, reduce, arrest, or ameliorate the growth or spread of cancer or tumors. In some embodiments, tumors are treated by arresting, reducing, or inhibiting further growth of the cancer or tumor. In some embodiments, the methods or uses described herein increase, enhance, or activate one or more immune responses to inhibit, reduce, arrest, or ameliorate the growth or spread of cancer or tumors. In some embodiments, the cancer or tumor reduces the size (e.g., volume or mass) of the cancer or tumor by at least 5%, at least 10%, at least 25%, at least 50%, or at least compared to the size of the cancer or tumor prior to treatment. It is treated by reducing it by 75%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%. In some embodiments, the cancer or tumor reduces the amount of cancer or tumor in the patient by at least 5%, at least 10%, at least 25%, at least 50%, at least 75%, or at least 90% compared to the amount of cancer or tumor prior to treatment. %, 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 about It represents a progression-free survival of 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 about Indicates an overall survival of 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 (ORR) of about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%. ).

Compounds and compositions according to the methods of the invention can be administered using any amount and any route of administration effective to inhibit MEK and treat or lessen the severity of a disease, such as those described herein. 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”, as used herein, refers to 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 can 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 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.

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, 19, 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 depends. 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 g/kg body weight/day of one or more other therapeutic agent(s) may be administered.

The amount of one or more other therapeutic agent(s) present in the compositions of the 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 ranges 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 TEAD inhibitors. In certain embodiments, the TEAD inhibitor is selected from those described in WO 2020/243415, the contents of which are incorporated herein by reference in their entirety. In certain embodiments, the TEAD inhibitor is selected from those described in WO 2020/243423, the contents of which are incorporated herein by reference in their entirety. In certain embodiments, the TEAD inhibitor is selected from those described in U.S. Patent 11,247,082, the contents of which are incorporated herein by reference in their entirety.

TEAD inhibitors can be produced by organic synthetic methods known to those skilled in the art. Additionally, specific TEAD inhibitors are described in Pobbati et al., “Targeting the Central Pocket in Human Transcription Factor TEAD as a Potential Cancer Therapeutic Strategy,” Structure 2015, 23, 2076-2086; Gibault et al., “Targeting Transcriptional Enhanced Associate Domains (TEADs),” J. Med. Chem. 2018, 61, 5057-5072; Bum-Erdene et al., “Small-Molecule Covalent Modification of Conserved Cysteine Leads to Allosteric Inhibition of the TEADoYap Protein-Protein Interaction,” Cell Chemical Biology 2019, 26, 1-12; Holden et al., “Small Molecule Dysregulation of TEAD Lipidation Induces a Dominant-Negative Inhibition of HippoPathway Signaling,” Cell Reports 2020, 31, 107809; WO 2017/053706, WO 2017/111076, WO 2018/204532, WO 2018/235926, US 20190010136, WO 2019/040380, WO 2019/113236, WO 2019/222431, WO 20 19/232216, WO 2020/051099, WO 2020 /081572, WO 2020/097389, WO 2020/190774, WO 2020/214734, PCT/US2020/35098, and PCT/US2020/35111, and the contents of each of the above documents and patent documents are as follows. Incorporated herein by reference in its entirety.

In some embodiments, the one or more other therapeutic agents are ERK5 inhibitors. In certain embodiments, the ERK5 inhibitor is selected from those described in WO 2022/051567, the contents of which are incorporated herein by reference in their entirety. In certain embodiments, the ERK5 inhibitor is selected from those described in WO 2022/051565, the contents of which are incorporated herein by reference in their entirety. In certain embodiments, the ERK5 inhibitor is selected from those described in WO 2022/051569, the contents of which are incorporated herein by reference in their entirety. In certain embodiments, the ERK5 inhibitor is selected from those described in WO 2022/051568, the contents of which are incorporated herein by reference in their entirety.

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 platin-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 platin-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. Approved VEGF inhibitors and kinase inhibitors useful in the present invention include: 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 some embodiments, 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-metabolite; platinous 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 PDGF 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 fusion products Compounds that target and reduce or inhibit the activity of, 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 factor Compounds that target, reduce or inhibit the activity of a family of receptors include, in particular, members of the EGF receptor tyrosine kinase family, such as compounds, proteins or antibodies that inhibit the EGF receptor, ErbB2, ErbB3 and ErbB4, or bind to EGF or an EGF-related ligand. , 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, but is not limited to, ABT-199, ABT-731, ABT-737, apogosypol, Ascenta's pan-Bcl-2 inhibitor, curcumin (and analogs thereof), Dual Bcl-2/Bcl-xL inhibitor (Infinity Pharmaceuticals/Novartis Pharmaceuticals), Genasense (G3139), HA14-1 (and its analogues; see WO2008118802), Navitoclax (and its analogues, see US7390799), NH- 1 (Shenayng Pharmaceutical University), obatoclax (and its analogs, see WO2004106328), S-001 (Gloria Pharmaceuticals), TW series compounds (Univ. of Michigan), and B-cell lymphoma 2, including venetoclax Includes compounds with inhibitory activity against protein (Bcl-2). 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, WO200612280 6, can be found in 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 “proteasome inhibitor” refers to a compound that targets, reduces or inhibits the activity of the proteasome. Compounds that target, reduce or inhibit the activity of the proteasome include, but are not limited to, bortezomib (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 immunity- tumor agent

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; WO14/03 6357) 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 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-oncology agent is an indoleamine (2,3)-dioxygenase (IDO) antagonist. In some embodiments, the IDO antagonist is epacadostat (INCB024360, Incyte); indoximod (NLG-8189, NewLink Genetics Corporation); Capmanitib (INC280, Novartis); GDC-0919 (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 contents of which are hereby incorporated by reference in their entirety. 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 the 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/B), an anti-KIR antibody, in leukemia (NCT01687387, NCT02399917, NCT02481297, NCT02599649), multiple myeloma (NCT02252263), and lymphoma (NCT01592370). ristol -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, but should not be construed as limiting it.

Example One: SPR black

MEK- RAF Combined Assay

Compound effects on the binding affinity of MEK to BRAF or CRAF are performed by surface plasmon resonance (SPR) using single cycle kinetic analysis. GST-BRAF or GST-CRAF is immobilized on the chip, and increasing concentrations of His-tagged MEK1 are flowed onto the sensor chip. Add 500 mM ATP to the running buffer. Dissociation constants for MEK1 binding to BRAF or CRAF are calculated in the absence and presence of compounds. The device is performed on a BIACORE 8K device. Sensorgrams are dual referenced and DMSO corrected. The data are fit to a single cycle kinetic model.

MEK- KSR Combined Assay

Compound effects on the binding affinity of MEK to KSR1 or KSR2 are performed by surface plasmon resonance (SPR) using single cycle kinetic analysis. GST-KSR1 or GST-KSR2 was immobilized on the chip, and increasing concentrations of His-tagged MEK1 were flowed onto the sensor chip. Add 500 mM ATP to the running buffer. Dissociation constants for MEK1 binding to BRAF or CRAF are calculated in the absence and presence of compounds. The device is performed on a BIACORE 8K device. Sensorgrams are dual referenced and DMSO corrected. The data are fit to a single cycle kinetic model.

MEK1 / CRAF stabilization

Anti-GST antibodies were immobilized on CM5 chips by amine coupling (GST capture kit from Cytiva). 1 μg/mL GST-CRAF protein (AA 306-548, N-terminal GST fusion, Carna Biosciences, Inc) was captured on this surface with a contact time of 200 s at a flow rate of 5 μL/min. Capture buffer was 1xPBS, pH 7.4, 10 mM MgCl ₂ , 1 mM DTT, 0.01% Tween-20, and 500 μM ATP. The capture level of CRAF was approximately 50 resonance units (RU).

His-tagged MEK1 (±compound) was diluted in buffer containing 10 mM HEPES, pH 7.4, 150 mM NaCl, 10 mM MgCl ₂ , 1 mM DTT, 0.01% Tween-20, 500 μM ATP, and 1% DMSO. . The running buffer was 1×PBS, 10 mM MgCl ₂ , 1 mM DTT, 0.01% Tween-20, 500 μM ATP, and 1% DMSO. MEK1 was serially diluted 3-fold starting from the highest dose of 200 nM using 3 μM compound solutions for a total of 5 doses. In each cycle, the association time was 60 seconds and the dissociation time was 600 seconds, with additional washing with 50% DMSO. Analyte binding was collected at 15°C using a flow rate of 30 μL/min.

The K _D ratio is defined as (K _D of the CRAF-MEK1 binary complex)/(K _D of the ternary complex).

Example 2: immunoprecipitation black

Immunoprecipitation experiments are performed by plating approximately 450,000 HCT116 cells per well in a 6-well plate. Cells are plated for 48 hours to reach approximately 70% confluence prior to transfection. Then, 24 hours after transfection, cells are treated with vehicle (0.1% DMSO) or compound (approximately 200 nM) for 1 hour. Cells are washed twice in cold PBS and then transferred to pre-chilled tubes in 0.6 ml of PBS solution. Cells are spun in a cold centrifuge at 1,800 g for 10 minutes, and the supernatant is aspirated. To lyse cells, the pellet was incubated in NP-40 buffer (50 mM Tris pH 7.8, 100 mM NaCl, 0.5% (v/v) NP-40, 10% NP-40) supplemented with protease and phosphatase inhibitor cocktail (Thermo Fisher, 78440). (v/v) glycerol, 1 mM EDTA) and incubate on ice for 30 minutes. Centrifuge the lysate at 2,100 g for 20 minutes and collect the supernatant. The clear lysate is quantified using BCA reagent (Pierce, 23225) with BSA as the standard. 5 μg of rabbit anti-MEK1 antibody (Millipore-Sigma, 07-641) or rabbit IgG (Millipore Sigma, 12-370) was immobilized on 50 μl of Sepharose Protein A Resin (Thermo Fisher, 53139), and immunoprecipitation was performed. Before starting, wash three times in 300 μl NP-40 buffer.

Next, for MEK1 immunoprecipitation, 250 μg of total cell lysate in a total volume of 0.6 ml is mixed with pre-immobilized anti-MEK1 antibody pre-conjugated to Protein A resin. Samples are incubated for 4 hours on an end-over-end rotator at 4°C and then washed three times in 0.6 ml volume of NP-40 buffer. Next, proteins are denatured and released from the resin by adding an 80 μl volume of 1× SDS sample buffer. Samples were boiled at 90°C for 2 min, spun, and then applied to a 4-12% bis-tris glycine gel (Bio-Rad, 3450125) run in MOPS-SDS buffer (Thermo Fisher, NP0001) at 150 V for 60 min. do. The gel is then transferred onto nitrocellulose in 20% methanol in Tris-glycine buffer (95 V, 250 A). Migration is confirmed using Ponceau red and then analyzed by Western blot. Signals for MEK, Flag-tagged proteins, BRAF, and GAPDH are detected by enhanced chemiluminescence on a ChemDoc XRS+ imaging system (Biorad).

Example 3: Biological assay to inhibit mouse xenograft tumor growth

Exemplary compounds with MEK inhibitory activity, alone or in combination with a second anticancer agent such as a TEAD inhibitor or an ERK5 inhibitor, can be tested for their ability to inhibit tumor growth in patient derived mouse xenograft models of pancreatic cancer and lung cancer. . The assay procedure is described below.

Patient-derived mouse xenograft ( PDX ) Procedure for tumor growth assay

Primary human tumor xenograft model tumor fragments ( ^2-3 mm in diameter) are inoculated subcutaneously into the right flank of Balb/c mice (6-8 weeks old) for tumor development. In one study, mice were inoculated with a human lung adenocarcinoma tumor model (MSCLC, ADC model LU6424) harboring a BRAF mutation (LU6424); In the second study, mice are inoculated with a human pancreatic tumor model (adenosquamous carcinoma model PA6258) harboring the K-Ras G12D mutation (PA6258). When the average tumor volume reaches approximately 150-200 mm ³ , animals are randomly assigned to the appropriate treatment group. Mice are treated with one of the following: (1) vehicle control, (2) an exemplary MEK inhibitor alone, (3) a second anti-cancer agent alone, or (4) a combination of a MEK inhibitor and a second anti-cancer agent. Tumors are measured twice per week using calipers.

Example 4: Biological assay

pERK HTRF black

Assay principle:

Human AsPc-1 cells or HCT-116 cells are seeded in 384-well culture plates and grown overnight. Cells are pretreated with compounds (10-pt titration) for 4 hours. Cell lysates are then prepared and the levels of phosphorylated ERK1/2 are analyzed using a homogeneous TR-FRET assay.

3D CTG viability test

Assay principle:

The CellTiter-Glo 3D cell viability assay is for determining cell viability in 3D cell spheroids. The assay reagent penetrates the large spheroid, and the dissolution capacity increases. This measures ATP as an indicator of viability and produces a luminescence reading. Cell Titer-Glo 3D cell viability assay kit Promega G9683 was used.

To NANOBRET General protocol for

Protein:protein interactions of MEK1 and BRAF, MEK1 and CRAF, MEK1 and KSR1, and MEK1 and KSR2 are assessed using the NANOBRET assay (Promega, Inc. Madison, Wisconsin). The full-length open reading frame of each gene pair is fused at the N-terminus or C-terminus with the open reading frame of the NANOLUC® or HALOTAG® coding sequence. Once the vectors are synthesized, the optimal signal window is determined through empirical evaluation of each pair of tag positions. The appropriate NANOLUC® and HALOTAG® chimeric gene fusions are transfected into HEK293 cells (or other cell lines) and cultured in white 96-well or 384-well plates suitable for tissue culture and signal assessment. Ikena compounds are distributed to the transfected live cells, along with necessary positive and negative controls. At separate times, NANOBRET™ NANOLUC® substrate is added and donor and acceptor signals are measured on the Envision Multimode (or other available instrument). Protein:protein interactions are assessed by calculating the NANOBRET™ ratio by subtracting the NANOBRET™ ratio of the control sample from the experimental sample. Additionally, expression of these chimeric gene fusions in mammalian, bacterial, or yeast protein expression systems can produce recombinant proteins that can be used in in vitro biochemical assays.

MEK1 Kinase assay protocol

Assay principle:

This is a binding assay involving activation of inactive ERK by MEK1 and phosphorylation of the ULIGHT-MBP substrate. The phosphorylated substrate was captured with an Eu-anti-phospho-substrate antibody that brought the Eu chelating donor and ULIGHT acceptor dye into close proximity. Upon excitation at 340 nm, the Eu chelate transfers its energy to the ULIGHT dye, producing fluorescence at 665 nm.

HCT116 ERK / pERK in-cell western assay

The In-Cell Western (ICW) assay is a quantitative immunofluorescence assay performed in microplates (optimized for 96-well or 384-well formats) that combines the specificity of Western blotting with the replicability and throughput of ELISA.

In-Cell Western assays are also called cytoblot, cell-based ELISA, In-Cell ELISA (ICE), and Fast Activated Cell-based ELISA (FACE). Using the In-Cell Western assay,

· ㆍ Detect proteins in fixed and permeabilized cultured cells using target-specific primary antibodies and IRDYE® secondary antibodies.

· ㆍ Quantify both targets at 700 nm and 800 nm using spectrally distinct infrared dye conjugates.

· ㆍ Quickly and accurately measures relative protein levels in many samples.

· ㆍ Detect proteins in situ in relevant cellular situations.

Tables 2 and 3 show the activity of selected compounds in the assays described above. Compound numbers correspond to those in Table 1 . Activity is classified as AD, where A indicates IC ₅₀ < 100 nM; B indicates 100 nM ≤ IC ₅₀ < 1 uM; C indicates 1 ≤ IC ₅₀ < 10 uM; D indicates IC ₅₀ ≥10 uM.

Table 4 shows the activity of selected compounds in the assay described above. Compound numbers correspond to those in Table 1 . Activity is classified as AD, where A indicates IC ₅₀ < 100 nM; B indicates 100 nM ≤ IC ₅₀ < 1 uM; C indicates 1 ≤ IC ₅₀ < 10 uM; D indicates IC ₅₀ ≥10 uM. CRaf/MEK SPR:KD ratio is classified as AD, where A indicates KD ratio <0.5; B indicates 0.5 ≤ KD ratio <1; C indicates 1 ≤ KD ratio <5; D indicates KD ratio ≥ 5.

Example 5: Synthesis of specific compounds

General synthesis scheme

General synthetic schemes for compounds as described herein are provided above. Additionally, synthetic procedures for certain exemplary compounds are provided below.

I-21

Step 1 : N -[2-(dimethylamino)ethyl] sulfamoyl chloride

N',N'-dimethylethane in DCM (5 mL) to a solution of sulfuryl chloride (1.53 g, 11.34 mmol, 1.13 mL, 1 eq) in DCM (50 mL) under N ₂ atmosphere at -40 to -50°C. -1,2-diamine (1 g, 11.34 mmol, 1.24 mL, 1 eq) and Et ₃ N (1.15 g, 11.34 mmol, 1.58 mL, 1 eq) are added dropwise. The mixture was stirred at -50°C for 5 minutes under N ₂ atmosphere to obtain N -[2-(dimethylamino)ethyl]sulfamoyl chloride (75 g, crude) as a white suspension, which was carried to the next step without further purification. used.

Step 2 : 3 - Cyclopropyl -1-[3-[2-(dimethylamino) ethylsulfamoylamino ]phenyl]-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl -Pyrido[4,3-d]pyrimidine-2,4,7-trione

1-(3-aminophenyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-pyrido[4] in DCM (10 mL) at 15°C. ,3-d]pyrimidine-2,4,7-trione (60 mg, 104.65 μmol, 100% purity, 1 eq) and Et ₃ N (72.70 mg, 718.46 μmol, 0.1 mL, 6.87 eq). N -[2-(dimethylamino)ethyl]sulfamoyl chloride (6 g, 32.14 mmol, not measurable (N/A) purity, 307.17 eq) was added. The mixture was stirred at 15°C for 5 minutes. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um; mobile phase: [water (0.05% NH ₃ H ₂ O + 10 mM NH ₄ HCO ₃ )-ACN]; B%: 48%-78%, 10 minutes) and lyophilized to obtain 3-cyclopropyl-1-[3-[2-(dimethylamino)ethylsulfamoylamino]phenyl]-5- (2-Fluoro-4-iodo-anilino)-6,8-dimethyl-pyrido[4,3-d]pyrimidine-2,4,7-trione (14.69 mg, 19.82 μmol, 18.9% Yield, 97.6% purity) was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.68 (dd, J = 1.8, 10.1 Hz, 1H), 7.58 (d, J = 8.4 Hz, 1H), 7.45-7.38 (m, 1H), 7.26-7.20 (m, 2H), 7.08 (d, J = 7.8 Hz, 1H), 6.88 (t, J = 8.5 Hz, 1H), 3.22 (s, 3H), 3.12 (t, J = 6.9 Hz, 2H), 2.73 (tt, J = 3.8, 7.2 Hz, 1H) , 2.51 (t, J = 6.9 Hz, 2H), 2.28 (s, 6H), 1.42 (s, 3H), 1.10-1.05 (m, 2H), 0.80-0.75 (m, 2H); ES-LCMS m/z 724.0 [M+H] ⁺ .

I-41 & I-42

Step 1 : tert -Butyl N -[2-( chlorosulfonylamino )ethyl] carbamate

Sulfuryl chloride (850 mg, 6.30 mmol, 629.63 μL, 1.01 eq) and Et ₃ N (1.89 g, 18.73 mmol, 2.61 mL, 3 eq) in DCM (50 mL) at -40 to -50°C under N ₂ atmosphere. To the solution of tert -butyl N -(2-aminoethyl)carbamate (1 g, 6.24 mmol, 980.39 μL, 1 eq) in DCM (5 mL) was added dropwise. The mixture was stirred for 5 minutes at -40 to -50°C under N ₂ atmosphere to obtain tert -butyl N -[2-(chlorosulfonylamino)ethyl]carbamate (75 g, crude) as a colorless liquid; This was used in the next step without further purification.

Step 2 : tert -Butyl N- [2-[[3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-2,4,7- trioxo-pyrido[4,3-d]pyrimidin-1-yl]phenyl]sulfamoylamino]ethyl]carbamate

1-(3-aminophenyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-pyrido[4,3-] in DCM (20 mL) d]pyrimidine-2,4,7-trione (90 mg, 156.97 μmol, 100% purity, 1 eq) and tert -butyl in a solution of Et ₃ N (145.40 mg, 1.44 mmol, 0.2 mL, 9.15 eq). N -[2-(chlorosulfonylamino)ethyl]carbamate (10 g, 38.65 mmol, not measurable purity, 246.23 eq) was added. The mixture was stirred at 15°C for 10 minutes. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was purified by flash silica gel chromatography (PE/EtOAc = 100/1 to 1/1, TLC: PE/EtOAc = 3/4, R _f = 0.48). Obtain the desired compound (100 mg). 50 mg of the desired compound (100 mg) was purified by preparative HPLC (column: Welch 10 minutes) and lyophilized to give tert -butyl N -[2-[[3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl- 2,4,7-trioxo-pyrido[4,3-d]pyrimidin-1-yl]phenyl]sulfamoylamino]ethyl]carbamate (16.07 mg, 20.07 μmol, 12.8% yield, 99.4% purity) was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.58 (dd, J = 1.8, 10.1 Hz, 1H), 7.49 (d, J = 8.2 Hz, 1H), 7.31 (t, J = 8.1 Hz, 1H), 7.17 (s, 1H), 7.13 (dd, J = 1.4, 8.1 Hz, 1H), 7.00-6.95 (m, 1H), 6.78 (t, J = 8.5 Hz, 1H), 3.13 (s, 3H), 3.03-2.98 (m, 2H), 2.97-2.92 ( m, 2H), 2.64 (tt, J = 3.8, 7.2 Hz, 1H), 1.33 (s, 9H), 1.32 (s, 3H), 1.02-0.96 (m, 2H), 0.71-0.66 (m, 2H) ; ES-LCMS m/z 796.1 [M+H] ⁺ .

Step 3 : 1 -[3-(2- Aminoethylsulfamoylamino )phenyl]-3- cyclopropyl -5-(2- fluoro- 4-iodo-anilino)-6,8-dimethyl-pyrido [4,3-d]pyrimidine-2,4,7-trione

tert -Butyl N- [2-[[3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-2,4 in DCM (2 mL) TFA in a solution of 7-trioxo-pyrido[4,3-d]pyrimidin-1-yl]phenyl]sulfamoylamino]ethyl]carbamate (50 mg, 62.84 μmol, not measurable purity, 1 eq) (3.08 g, 27.01 mmol, 2 mL, 429.83 eq) was added. The mixture was stirred at 20°C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um; mobile phase: [water (0.05% NH ₃ H ₂ O + 10 mM NH ₄ HCO ₃ )-ACN]; B%: 45%-75%, 10 minutes) and lyophilized to obtain 1-[3-(2-aminoethylsulfamoylamino)phenyl]-3-cyclopropyl-5-(2- Fluoro-4-iodo-anilino)-6,8-dimethyl-pyrido[4,3-d]pyrimidine-2,4,7-trione (16.78 mg, 23.94 umol, 38.09% yield, 99.223 % purity) was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.68 (dd, J = 1.8, 10.1 Hz, 1H), 7.59 (d, J = 8.4 Hz, 1H), 7.41 (t, J = 8.0 Hz, 1H), 7.28-7.25 (m, 1H), 7.25-7.22 (m, 1H), 7.08-7.04 (m, 1H), 6.88 (t, J = 8.5 Hz, 1H), 3.22 (s, 3H), 3.06 (t, J = 6.1 Hz, 2H), 2.76-2.70 ( m, 3H), 1.41 (s, 3H), 1.10-1.05 (m, 2H), 0.80-0.76 (m, 2H); ES-LCMS m/z 696.0 [M+H] ⁺ .

I-24

Step 1 : 3 - Cyclopropyl -5-((2- fluoro- 4- iodophenyl )amino)-1-(3- hydroxyphenyl )-6,8-dimethylpyrido[4,3- d ] Pyrimidine-2,4,7(1 H ,3 H ,6H)-trione

An aqueous solution of NaNO ₂ (9.63 mg, 139.53 μmol, 2 eq) in H ₂ O (1 mL) was reacted with 1-(3-aminophenyl) in H ₂ SO ₄ (0.25 mL) and H ₂ O (0.5 mL) at 0°C. )-3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethylpyrido[4,3- d ]pyrimidine-2,4,7(1 H ,3 H ,6 H )-trione (40 mg, 69.76 μmol, 100.0% purity, 1 eq) was added to the solution. The reaction mixture was stirred for 30 minutes. AcOH (2 mL) was added. The reaction mixture was stirred at 100°C for 2 hours. The reaction mixture was concentrated under reduced pressure to remove most of the acetic acid. To the residue was added saturated aqueous NaHCO ₃ to pH 7 and extracted using EtOAc (20 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 preparative TLC (PE/EtOAc = 1/1, R _f = 0.4) to give 3-cyclopropyl- 5-((2-fluoro-4-iodophenyl)amino)-1-(3-hydroxyphenyl)-6,8-dimethylpyrido[4,3- d ]pyrimidine-2,4,7 ( 1H , 3H ,6H)-trione (35 mg, 55.45 μmol, 79.5% yield, 91.0% purity) was obtained as a white solid. ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 11.30 (s, 1H), 7.53 (d, J = 9.3 Hz, 1H), 7.46 (d, J = 8.4 Hz, 1H), 7.29 (s, 1H), 6.84 (d, J = 7.8 Hz, 2H), 6.80 (s, 1H), 6.70 (t, J = 8.0 Hz, 1H), 5.59 (s, 1H), 3.21 (s, 3H), 2.75 (s, 1H) ), 1.44 (s, 3H), 1.16-1.10 (m, 2H), 0.86 (d, J = 16.8 Hz, 2H); ES-LCMS m/z 574.8 [M+H] ⁺ .

Step 2 : 3 -(3- cyclopropyl- 5-((2- fluoro -4- iodophenyl )amino)-6,8-dimethyl-2,4,7-trioxo-3,4,6, 7-Tetrahydropyrido[4,3- d ]pyrimidin-1(2H ) -yl)phenyl methylsulfamate

3-Cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-1-(3-hydroxyphenyl)-6,8-dimethylpyrido[4,3] in DCM (1 mL) -d]pyrimidine-2,4,7(1 H ,3 H ,6 H )-trione (25 mg, 55.45 μmol, 79.5% yield, 91.0% purity, 1 eq) in N -methylsulfamoyl Chloride (4.62 mg, 35.65 μmol, 0.9 eq) and Et ₃ N (12.02 mg, 118.83 μmol, 16.54 μL, 3 eq) were added. The mixture was stirred at 25°C for 1 hour. The reaction mixture was acidified by adding 0.1 N HCl and extracted using EtOAc (20 mL x 3). The organic layer was washed with brine (20 mL x 2), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um; mobile phase: [water (0.05% NH ₃ H ₂ O + 10 mM NH ₄ HCO ₃ )-ACN]; B%: 55% -85%, 10 min) and lyophilization to obtain the desired compound (20 mg, purity 60.0%) as a white solid, which was further purified by preparative TLC (PE/EtOAc = 1/1, R _f = 0.38). Purified to obtain 3-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo-3,4,6,7 -Tetrahydropyrido[4,3- d ]pyrimidin-1( 2H )-yl)phenyl methylsulfamate (8.34 mg, 12.50 μmol, 31.6% yield, 100.0% purity) was obtained as a white solid. ^1H NMR (500 MHz, CDCl ₃ ) δ ppm 11.28 (s, 1H), 7.53 (dd, J = 1.7, 9.6 Hz, 1H), 7.49-7.45 (m, 2H), 7.34-7.29 (m, 2H) , 7.25 (d, J = 2.3 Hz, 1H), 6.72 (t, J = 8.3 Hz, 1H), 4.80 (s, 1H), 3.21 (s, 3H), 2.96-2.91 (m, 3H), 2.80- 2.71 (m, 1H), 1.40 (s, 3H), 1.16-1.11 (m, 2H), 0.83-0.77 (m, 2H); ES-LCMS m/z 668.0 [M+H] ⁺ .

I-25

Step 1 : 2 -Methylbenzene-1,3- diamine

Pd/C (1 g, 10.98 mmol, 10.0% purity, 1.00%) in a solution of 2-methyl-1,3-dinitro-benzene (2 g, 10.98 mmol, 1 eq) in MeOH (15 mL) under Ar ₂ atmosphere. eq) was added. The suspension was degassed under vacuum and purged several times with H ₂ . The mixture was stirred at 25° C. under H ₂ (30 psi) for 1 hour. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue, which was subjected to flash silica gel chromatography (PE/EtOAc = from 100/1 to 1/1, TLC: PE/EtOAc = 1/1, R _f = 0.37) Purification was performed to obtain 2-methylbenzene-1,3-diamine (800 mg, 6.35 mmol, 57.9% yield, 97.0% purity) as a gray solid. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 7.67-7.15 (m, 1H), 6.89-6.81 (m, 2H), 6.22 (d, J = 7.8 Hz, 4H), 1.99 (s, 3H).

Step 2 : [3- Cyclopropyl -1-(2- Fluoro- 4- iodo -phenyl)-6,8-dimethyl-2,4,7- trioxo -pyrido[2,3- d ]pyri midin-5-yl] 4- methylbenzenesulfonate

3-Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-5-hydroxy-6,8-dimethyl-pyrido[2,3- d ]pyrimidine- in MeCN (35 mL) A solution of 2,4,7-trione (800 mg, 1.66 mmol, 100.0% purity, 1 eq) was added with 4-methylbenzenesulfonyl chloride (479.74 mg, 2.52 mmol, 1.52 eq) and Et ₃ N (581.30 mg, 5.74 mmol, 799.58 μL, 3.47 eq) was added. The mixture was stirred at 15°C for 2 hours under N ₂ atmosphere. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was purified by flash silica gel chromatography (PE/EtOAc = 100/1 to 1/1, TLC: PE/EtOAc = 2/1, R _f = 0.3). [3-Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2,3- d ]pyrimidine-5 -yl] 4-methylbenzenesulfonate (620 mg, 924.04 μmol, 55.8% yield, 95.0% purity) was obtained as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 11.28 (s, 1H), 7.53 (d, J = 9.0 Hz, 1H), 7.47 (s, 2H), 7.37-7.29 (m, 2H), 7.26-7.23 (m, 1H), 6.72 (t, J = 7.2 Hz, 1H), 4.95 (s, 1H), 3.21 (s, 3H), 2.93 (s, 3H), 2.74 (s, 1H), 1.40 (s, 3H), 1.27 (s, 2H), 1.14 (d, J = 6.1 Hz, 2H); ES-LCMS m/z 638.0 [M+H] ⁺ .

Step 3 : 5 -(3-Amino-2- methyl - anilino )-3- cyclopropyl -1-(2- fluoro- 4- iodo -phenyl)-6,8-dimethyl-pyrido[2, 3- d ]pyrimidine-2,4,7-trione

[3-Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2,3-] in DMA (2 mL) d ]pyrimidin-5-yl] 2-methylbenzene-1,3-diamine (123.89 mg, 983.66 μmol, 97.0 μmol) in a solution of 4-methylbenzenesulfonate (220 mg, 327.89 μmol, 95.0% purity, 1 eq). % purity, 3 eq) and 2,6-lutidine (105.40 mg, 983.66 μmol, 114.56 μL, 3 eq) were added. The mixture was stirred at 180° C. under a microwave for 2 hours. The mixture was diluted with water (20 mL) and extracted using ethyl acetate (30 mL x 3). The organic layer was dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (0.05% HCl) )-ACN]; B%: 46%-76%, 10 minutes) and lyophilized to obtain 5-(3-amino-2-methyl-anilino)-3-cyclopropyl-1-(2-fluoro -4-iodo-phenyl)-6,8-dimethyl-pyrido [2,3- d ] pyrimidine-2,4,7-trione (58 mg, 88.65 μmol, 27.0% yield, 95.4% purity, HCl) was obtained as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 10.16 (s, 1H), 7.70-7.59 (m, 2H), 7.14 (d, J = 7.8 Hz, 2H), 7.08-6.77 (m, 1H), 6.67 (d, J = 6.1 Hz, 1H), 2.97 (s, 3H), 2.78 (s, 1H), 2.55 (s, 3H), 1.60 (s, 3H), 1.18 (d, J = 5.6 Hz, 2H) , 0.83 (s, 2H); ES-LCMS m/z 588.1 [M+H] ⁺ .

Step 4 : 3 - Cyclopropyl -1-(2- Fluoro- 4 - iodo -phenyl)-6,8-dimethyl-5- [2-methyl-3-(methylsulfamoylamino)anilino] pyrido [2,3- d ]pyrimidine-2,4,7-trione

5-(3-Amino-2-methyl-anilino)-3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-pyrido in DCM (1 mL) [2,3- d ]pyrimidine-2,4,7-trione (50 mg, 76.42 μmol, 95.4% purity, 1 eq, HCl) was added with N -methylsulfamoyl chloride (8.91 mg, 68.78 μmol, 0.9 eq) and TEA (38.66 mg, 382.11 μmol, 53.18 μL, 5 eq) were added. The mixture was stirred at 5°C for 1 hour under N ₂ atmosphere. To the residue was added saturated aqueous NaHCO ₃ to pH 7 and extracted using EtOAc (20 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 prep-TLC (SiO ₂ , PE/EtOAc = 1/1, R _f = 0.38) to give 3 -Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-5-[2-methyl-3-(methylsulfamoylamino)anilino]pyrido[2,3 - d ]pyrimidine-2,4,7-trione (50 mg, 70.19 μmol, 91.9% yield, 95.5% purity) was obtained as a white solid. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 10.11 (s, 1H), 7.63 (d, J = 9.5 Hz, 1H), 7.22 (d, J = 8.1 Hz, 1H), 7.12 (t, J = 7.7) Hz, 1H), 6.95 (s, 1H), 6.67 (d, J = 7.6 Hz, 1H), 6.25 (s, 1H), 4.50 (s, 1H), 4.12 (s, 1H), 2.96 (s, 3H) ), 2.85 (s, 1H), 2.79 (s, 3H), 2.74 (d, J = 4.6 Hz, 3H), 1.54 (s, 3H), 1.18 (d, J = 6.1 Hz, 2H), 0.83 (d) , J = 5.4 Hz, 2H); ES-LCMS m/z 681.0 [M+H] ⁺ .

Step 5 : 3 - Cyclopropyl -5- (2- Fluoro -4- Iodo - Anilino ) -6,8-dimethyl-1- [2-methyl-3- (methylsulfamoylamino) phenyl] pyrido [4,3- d ]pyrimidine-2,4,7-trione

3-Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-5-[2-methyl-3-(methylsulfamoylamino)anilino in THF (1 mL) ]pyrido[2,3- d ]pyrimidine-2,4,7-trione (30 mg, 42.11 μmol, 95.5% purity, 1 eq) was added to a solution of NaOMe/MeOH (28 wt%, 42.11 μmol, 1 eq). eq) One drop was added. The mixture was stirred at 25°C for 1 hour. AcOH was added to the residue until pH 7 and extracted using EtOAc (20 mL x 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (0.05% HCl)-ACN]; B%: 50%-70%, 10 minutes) and frozen. Drying gave the product, to which was added saturated aqueous NaHCO ₃ until pH = 7, extracted using EtOAc (20 mL x 3), then the combined organic layers were dried over Na ₂ SO ₄ , filtered and reduced pressure. Concentrated under 3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-1-[2-methyl-3-(methylsulfamoylamino)phenyl]pyrido [4,3- d ]pyrimidine-2,4,7-trione (10.19 mg, 14.66 μmol, 34.8% yield, 97.9% purity) was obtained as a white solid. ^1H NMR (400 MHz, CD ₃ OD) δ ppm 7.70-7.63 (m, 1H), 7.55 ( J = 8.4, 12.8 Hz, 2H), 7.28 ( J = 7.8 Hz, 1H), 7.06 (d, J = 7.1 Hz, 1H), 6.86 (t, J = 8.6 Hz, 1H), 3.20 (s, 3H), 2.74-2.69 (m, 1H), 2.67 (s, 3H), 2.24 (s, 3H), 1.27 ( s, 3H), 1.07 (d, J = 7.1 Hz, 2H), 0.89-0.76 (m, 2H); ES-LCMS m/z 681.0 [M+H] ⁺ .

I-29

Step 1 : 1 -[3-[3- Cyclopropyl -5- (2- Fluoro -4- Iodo - Anilino ) -6,8-dimethyl-2,4,7-trioxo-pyrido [4 ,3-d]pyrimidin-1-yl]phenyl]-3-(methylsulfamoyl)urea

1-(3-aminophenyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl in THF (20 mL) at -60°C under N ₂ atmosphere. -N -(oxomethylene)sulfamoyl chloride (100 mg, 706.55 μmol, 61.35 μL, 2.03 eq) was added. The mixture was stirred at -60 to -20°C for 0.5 hours under N ₂ atmosphere. MeNH ₂ (235 mg, 3.48 mmol, 100% purity, 9.98 eq, HCl) and Et ₃ N (352.97 mg, 3.49 mmol, 485.52 μL, 10 eq) were added. The mixture was slowly warmed to 25°C and stirred at 25°C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (0.05% HCl)-ACN]; B%: 50% -70%, 10 minutes) and lyophilized. The residue was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um; mobile phase: [water (0.05% NH ₃ H ₂ O + 10 mM NH ₄ HCO ₃ )-ACN]; B%: 25% -55%, 10 minutes) and lyophilized to obtain 1-[3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-2,4. ,7-trioxo-pyrido[4,3-d]pyrimidin-1-yl]phenyl]-3-(methylsulfamoyl)urea (18.94 mg, 25.86 μmol, 7.4% yield, 96.9% purity) as white Obtained as a solid. ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 7.70-7.52 (m, 3H), 7.40 (d, J = 4.6 Hz, 2H), 7.08 (d, J = 3.2 Hz, 1H), 6.86 (t, J = 8.6 Hz, 1H), 3.20 (s, 3H) ), 2.75-2.65 (m, 4H), 1.40 (s, 3H), 1.10-1.00 (m, 2H), 0.80-0.70(m, 2H); ES-LCMS m/z 710.0 [M+H] ⁺ .

I-44

Step 1 : N -[3-[6- Cyclopropyl -4- (2- Fluoro -4- Iodo - Anilino ) -3,7-dimethyl-2,5-dioxo-pyrano [3,2 -c]pyridin-8-yl]- 2- methyl -phenyl] acetamide

8-(3-Amino-2-methyl-phenyl)-6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-3,7-dimethyl-pyrano in DCM (2 mL) Acetyl chloride ( 4.95 mg, 63.01 μmol, 4.50 μL, 2 eq) was added. The reaction mixture was stirred at 0° C. for 1 hour under N ₂ atmosphere. The reaction mixture was quenched by adding saturated aqueous NaHCO ₃ (15 mL) at 0° C. and extracted using DCM (20 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 analyzed by preparative TLC (PE/EtOAc = 0/1, TLC: PE/ EtOAc = 0/1, R _f = 0.45) and then lyophilized to obtain N -[3-[6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-3,7-dimethyl-2,5-dioxo -Pyrano[3,2-c]pyridin-8-yl]-2-methyl-phenyl]acetamide (13.71 mg, 22.31 μmol, 70.8% yield, 99.8% purity) was obtained as a white solid. ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 11.32 (s, 1H) _, 9.40 (s, 1H), 7.72 (d, J = 10.3 Hz, 1H), 7.52 (d, J = 8.3 Hz, 2H) ), 7.26 (t, J = 7.6 Hz, 1H), 7.01 (d, J = 7.6 Hz, 1H), 6.88 (t, J = 8.7 Hz, 1H), 3.04 (d, J = 4.2 Hz, 1H), 2.24 (s, 3H), 2.08 (s, 3H), 1.95 (s, 3H), 1.45 (s, 3H), 1.21 (d, J = 8.1 Hz, 2H), 0.97-0.88 (m, 2H); ES-LCMS m/z 614.0 [M+H] ⁺ .

I-45

Step 1 : 2 -(3- nitrophenyl )-2-oxo-acetic acid

To a solution of 2-oxo-2-phenyl-acetic acid (51 g, 339.70 mmol, 1 eq) in H ₂ SO ₄ (80 mL) at 0° C. was added KNO ₃ (41.21 g, 407.64 mmol, 1.2 eq). The mixture was stirred at 25°C for 16 hours. The reaction mixture was poured into ice water (1.6 L) and extracted using EtOAc (400 mL x 3). The combined organic phases were washed with brine (300 mL), dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give 2-(3-nitrophenyl)-2-oxo-acetic acid (66 g, crude). was obtained as a yellow solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.70 (t, J = 1.8 Hz, 1H), 8.56 (td, J = 1.2, 8.2 Hz, 1H), 8.41-8.37 (m, 1H), 7.89 (t, J = 8.0 Hz, 1H).

Step 2 : Ethyl 2-(3- nitrophenyl )-2-oxo-acetate

To a solution of 2-(3-nitrophenyl)-2-oxo-acetic acid (66 g, 338.24 mmol, 1 eq) in DMF (700 mL) was added K ₂ CO ₃ (121.54 g, 879.42 mmol, 2.6 eq). . After stirring at 20°C for 2 hours, iodoethane (263.77 g, 1.69 mol, 135.26 mL, 5 eq) was added dropwise. The mixture was stirred at 20°C for 16 hours. The reaction mixture was poured into water (800 mL) and extracted using EtOAc (500 mL x 3). The combined organic layers were washed with brine (500 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 (PE/EtOAc = 1/0 to 10/ 1, purified by TLC: PE/EtOAc = 5/1, R _f = 0.45) to give ethyl 2-(3-nitrophenyl)-2-oxo-acetate (35.5 g, 143.16 mmol, 42.3% yield, 90.0% purity). ) was obtained as a yellow oil. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 8.90 (s, 1H), 8.54-8.47 (m, 1H), 8.40 (d, J = 7.8 Hz, 1H), 7.75 (t, J = 7.9 Hz, 1H) ), 4.50 (q, J = 7.2 Hz, 2H), 1.46 (t, J = 7.1 Hz, 3H).

Step 3 : Ethyl 3-[(2 E )-2-[2 -Ethoxy -1-(3- nitrophenyl )-2-oxo- ethylidene ] hydrazino ]-3-oxo-propanoate

To a solution of ethyl 2-(3-nitrophenyl)-2-oxo-acetate (22.5 g, 90.73 mmol, 1 eq) in EtOH (120 mL) was added H ₂ SO ₄ (2.29 g, 23.33 mmol, 1.24 mL, 0.25 eq). ) and ethyl 3-hydrazino-3-oxo-propanoate (13.79 g, 94.36 mmol, 1.04 eq) were added. The mixture was stirred at 90°C for 2 hours. The solvent was removed to give a residue, which was diluted with water (200 mL) and extracted using EtOAc (300 mL x 3). The combined organic phases were washed with brine (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give ethyl 3-[(2 E )-2-[2-ethoxy-1-(3). -Nitrophenyl)-2-oxo-ethylidene]hydrazino]-3-oxo-propanoate (30 g, crude, E/Z mixture) was obtained as a yellow oil. ES-LCMS m/z 352.2 [M+H] ⁺ .

Step 4 : Ethyl 4- hydroxy -3-(3- nitrophenyl )-6-oxo- 1H - pyridazine -5- carboxylate

Ethyl 3-[(2 E )-2-[2-ethoxy-1-(3-nitrophenyl)-2-oxo-ethylidene]hydrazino]-3-oxo-propano in DMF (300 mL) To a solution of ate (30 g, 85.39 mmol, E/Z mixture, 1 eq) was added K ₂ CO ₃ (6.49 g, 46.97 mmol, 0.55 eq). The mixture was stirred at 80°C for 3 hours. The mixture was cooled to ambient temperature and poured into 3N HCl (600 mL). The precipitated solid was collected by filtration to obtain a residue, to which MeOH (50 mL) was added and stirred at 15°C for 2 hours. The slurry was filtered and the cake was rinsed with MeOH (3 x 3 mL). The solid was collected and dried in vacuo to obtain ethyl 4-hydroxy-3-(3-nitrophenyl)-6-oxo-1 H -pyridazine-5-carboxylate (8.3 g, 24.47 mmol, 28.7% yield, 90.0 % purity) was obtained as a yellow solid. ^1H NMR (400MHz, DMSO- d6 ) δ ppm 13.23 (s, 1H), 8.58-8.53 (m, 1H), 8.30 (td, J = 1.2, _7.2 Hz, 1H), 8.15 (d, J = 8.1) Hz, 1H), 7.76 (t, J = 8.1 Hz, 1H), 4.32 (q, J = 7.1 Hz, 2H), 1.29 (t, J = 7.1 Hz, 3H); ES-LCMS m/z 306.1 [M+H] ⁺ .

Step 5 : Ethyl 2-allyl-5- hydroxy -6- (3- nitrophenyl ) -3-oxo- pyridazine -4- carboxylate

To a solution of ethyl 4-hydroxy-3-(3-nitrophenyl)-6-oxo-1H-pyridazine-5-carboxylate (3 g, 8.85 mmol, 1 eq) in DMF (40 mL) was added NaH (707.63). mg, 17.69 mmol, 60.0% purity, 2 eq) was added and stirred at 20°C for 30 minutes. The solution was cooled to -10°C and 3-bromoprop-1-ene (1.12 g, 9.29 mmol, 1.05 eq) was added dropwise. The reaction mixture was slowly warmed to 20°C and stirred at 20°C for 1 hour. The reaction mixture was quenched by addition of water (60 mL) and extracted using EtOAc (80 mLx3). The combined organic layers were washed with brine (50 mL), dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give ethyl 2-allyl-5-hydroxy-6-(3-nitrophenyl)-3-oxo. -Pyridazine-4-carboxylate (3 g, 7.82 mmol, 88.4% yield, 90.0% purity) was obtained as a yellow oil. ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 8.57 (s, 1H), 8.30 (d, J = 1.5, _8.3 Hz, 1H), 8.16 (d, J = 7.8 Hz, 1H), 7.76 (t) , J = 8.1 Hz, 1H), 5.98 (td, J = 5.5, 10.9, 16.8 Hz, 1H), 5.24-5.15 (m, 2H), 4.70 (d, J = 5.6 Hz, 2H), 4.30 (q, J = 7.1 Hz, 2H), 1.28 (t, J = 7.1 Hz, 3H); ES-LCMS m/z 346.0 [M+H] ⁺ .

Step 6 : Ethyl 2-allyl-5- chloro -6- (3- nitrophenyl ) -3-oxo- pyridazine -4- carboxylate

To a solution of ethyl 2-allyl-5-hydroxy-6-(3-nitrophenyl)-3-oxo-pyridazine-4-carboxylate (3 g, 7.82 mmol, 1 eq) in DCM (40 mL) ( COCl)₂(9.92 g, 78.19 mmol, 6.84 mL, 10 eq) was added. The mixture was stirred at 35°C for 3 hours. Removal of the solvent gave ethyl 2-allyl-5-chloro-6-(3-nitrophenyl)-3-oxo-pyridazine-4-carboxylate (3 g, crude) as a yellow solid.^OneH NMR (400 MHz, CDCl₃)δ ppm 8.46 (t,J= 1.8 Hz, 1H), 8.37-8.33 (m, 1H), 7.91 (d,J= 1.0, 7.8 Hz, 1H), 7.68 (t,J= 7.9 Hz, 1H), 6.01 (td,J= 6.2, 10.3, 16.9 Hz, 1H), 5.41-5.27 (m, 2H), 4.82 (d,J= 6.4 Hz, 2H), 4.49 (q,J= 7.2 Hz, 2H), 1.42 (t,J= 7.2 Hz, 3H); ES-LCMSm/z 364.1, 366.1 [M+H]⁺.

Step 7 : Ethyl 2-allyl-5-( methylamino )-6-(3- nitrophenyl )-3-oxo- pyridazine -4- carboxylate

A solution of ethyl 2-allyl-5-chloro-6-(3-nitrophenyl)-3-oxo-pyridazine-4-carboxylate (3 g, 6.60 mmol, 1 eq) in DCM (30 mL) at 0°C. MeNH ₂ (13.66 g, 131.96 mmol, 30.0% purity, 20 eq) was added. The mixture was stirred at 0°C for 5 hours. The reaction mixture was quenched by adding water (50 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 (PE/EtOAc = 1/1, TLC: PE/EtOAc = 1/1, R _f = 0.24) to give ethyl 2-allyl-5-(methylamino)-6-(3-nitrophenyl)-3-oxo-pyridazine-4-carboxylate (2.2 g, 5.53 mmol, 83.7% yield, 90.0% purity) was obtained as a yellow solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.38-8.32 (m, 1H), 8.29 (t, J = 1.7 Hz, 1H), 7.90 (d, J = 7.8 Hz, 1H), 7.83-7.76 (m, 1H), 6.24 (d, J = 5.1 Hz, 1H), 5.98-5.84 (m, 1H), 5.21-5.07 (m, 2H), 4.56 (d, J = 5.6 Hz, 2H), 4.24 ( q, J = 7.1 Hz, 2H), 2.66 (d, J = 5.1 Hz, 3H), 1.28 (t, J = 7.1 Hz, 3H); ES-LCMS m/z 359.1 [M+H] ⁺ .

Step 8 : 2 -allyl-5-( methylamino )-6-(3- nitrophenyl ) pyridazin -3-one

A solution of ethyl 2-allyl-5-(methylamino)-6-(3-nitrophenyl)-3-oxo-pyridazine-4-carboxylate (1.5 g, 3.77 mmol, 1 eq) in DMSO (20 mL) LiCl (3.19 g, 75.35 mmol, 20 eq) was added. The mixture was stirred at 180°C for 6 hours. The mixture was diluted with water (40 mL) and extracted using EtOAc (60 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 (PE/EtOAc = from 1/0 to 1/2, TLC: PE/EtOAc = 1/2, R _f = 0.25) to obtain 2-allyl-5-(methylamino)-6-(3-nitrophenyl)pyridazin-3-one (1.2 g, 3.35 mmol, 89.0% yield, 80.0%) Purity) was obtained as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 8.38 (d, J = 1.7 Hz, 1H), 8.34-8.29 (m, 1H), 7.85 (d, J = 7.6 Hz, 1H), 7.70 (t, J = 7.9 Hz, 1H), 6.09-5.94 (m, 1H), 5.86 (s, 1H), 5.28-5.17 (m, 2H), 4.73 (d, J = 4.6 Hz, 2H), 4.29 (s, 1H) , 2.85-2.79 (m, 3H); ES-LCMS m/z 287.1 [M+H] ⁺ .

Step 9 : 6 -allyl-4- hydroxy -1,3-dimethyl-8- (3-nitrophenyl) pyrido [2,3-d] pyridazine -2,5-dione

2-methylpropanedioic acid in a solution of 2-allyl-5-(methylamino)-6-(3-nitrophenyl)pyridazin-3-one (1.2 g, 3.35 mmol, 1 eq) in AcO (20 mL) (1.19 g, 10.06 mmol, 3 eq) was added. The mixture was stirred at 110°C for 2 hours. The solvent was removed to obtain a residue, which was purified by preparative TLC (PE/EtOAc = 1/1, TLC: PE/EtOAc = 1/1, Rf = 0.40) and then lyophilized to give 6-allyl-4- Hydroxy-1,3-dimethyl-8-(3-nitrophenyl)pyrido[2,3-d]pyridazine-2,5-dione (1.1 g, 2.45 mmol, 73.0% yield, 82.0% purity) Obtained as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.50-8.27 (m, 2H), 7.83-7.60 (m, 2H), 6.10-5.92 (m, 1H), 5.39-5.31 (m, 2H), 4.86 ( d, J = 6.1 Hz, 2H), 3.08 (s, 3H), 2.15 (s, 3H); ES-LCMS m/z 369.1 [M+H] ⁺ .

Step 10 : [6-allyl-1,3-dimethyl-8-(3- nitrophenyl )-2,5- dioxo - pyrido[2,3-d]pyridazin -4-yl] 4- methylbenzene sulfonate

6-allyl-4-hydroxy-1,3-dimethyl-8-(3-nitrophenyl)pyrido[2,3-d]pyridazine-2,5-dione (1.1 g, To a solution of 2.45 mmol, 1 eq), DIEA (1.58 g, 12.24 mmol, 2.13 mL, 5 eq) and 4-methylbenzenesulfonyl chloride (933.71 mg, 4.90 mmol, 2 eq) were added. The mixture was stirred at 60°C for 1 hour. The mixture was diluted with water (50 mL) and extracted using EtOAc (60 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 (PE/EtOAc = from 1/0 to 1/1, TLC: PE/EtOAc = 1/1, R _f = 0.35) to obtain [6-allyl-1,3-dimethyl-8-(3-nitrophenyl)-2,5-dioxo-pyrido[2,3-d]pyridazine. -4-yl] 4-methylbenzenesulfonate (1.2 g, 1.88 mmol, 76.9% yield, 82.0% purity) was obtained as a brown solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.41-8.30 (m, 2H), 7.97 (d, J = 8.3 Hz, 2H), 7.79-7.66 (m, 2H), 7.41 (d, J = 8.3 Hz) , 2H), 6.10-5.94 (m, 1H), 5.40-5.22 (m, 2H), 4.84 (d, J = 6.1 Hz, 2H), 3.09 (s, 3H), 2.50 (s, 3H), 1.89 ( s, 3H); ES-LCMS m/z 523.1 [M+H] ⁺ .

Step 11 : 6 -allyl-4-(2- fluoro- 4 - iodo - anilino )-1,3-dimethyl-8- (3-nitrophenyl)pyrido[2,3-d] pyridazine- 2,5-dione

To a solution of 2-fluoro-4-iodo-aniline (647.80 mg, 2.73 mmol, 2.49 eq) in THF (10 mL) was added NaH (175.74 mg, 4.39 mmol, 60.0% purity, 4 eq), Stir at ℃ for 30 minutes, and stir at 0℃ for [6-allyl-1,3-dimethyl-8-(3-nitrophenyl)-2,5-dioxo-pyrido[2,3-d]pyridazine- 4-day] 4-methylbenzenesulfonate (700 mg, 1.10 mmol, 1 eq) was added. The reaction temperature was slowly warmed to 25°C and stirred at 25°C for 3 hours. The reaction mixture was quenched by adding water (30 mL) and extracted using EtOAc (40 mL x 3). 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 (PE/EtOAc = 100/1 to 3/ 1, purified by TLC: PE/EtOAc = 3/1, R _f = 0.43) to give 6-allyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-8- (3-Nitrophenyl)pyrido[2,3-d]pyridazine-2,5-dione (150 mg, 217.08 μmol, 19.7% yield, 85.0% purity) was obtained as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 10.72 (s, 1H), 8.42-8.39 (m, 1H), 8.36 (td, J = 1.8, 7.8 Hz, 1H), 7.77-7.66 (m, 2H) , 7.46 (dd, J = 1.8, 10.0 Hz, 1H), 7.38 (d, J = 8.7 Hz, 1H), 6.57 (t, J = 8.5 Hz, 1H), 6.04 (td, J = 6.1, 10.4, 16.9 Hz, 1H), 5.39-5.28 (m, 2H), 4.86 (d, J = 6.0 Hz, 2H), 3.08 (s, 3H), 1.82 (s, 3H); ES-LCMS m/z 588.0 [M+H] ⁺ .

Step 10 : 6 -Allyl-8-(3- aminophenyl )-4-(2- fluoro- 4 - iodo - anilino )-1,3-dimethyl-pyrido[2,3-d]pyridazine -2,5-dione

6-allyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-8-(3-nitrophenyl)pyri in EtOH (1.5 mL) and H ₂ O (1.5 mL) [2,3-d]pyridazine-2,5-dione (130 mg, 188.14 μmol, 1 eq) was added to a solution of NH ₄ Cl (100.64 mg, 1.88 mmol, 10 eq) and Fe (52.53 mg, 940.68 μmol). , 5 eq) was added. The mixture was stirred at 90°C for 3 hours. The reaction mixture was filtered to remove insoluble material. The filtrate was concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: AgNO ₃ _ silica 150*25 mm*15 um; mobile phase: [heptane-EtOH (0.1% NH ₃ H ₂ O)]; B %: 0%-80%, 9 minutes) and then lyophilized to produce 6-allyl-8-(3-aminophenyl)-4-(2-fluoro-4-iodo-anilino)-1. ,3-Dimethyl-pyrido[2,3-d]pyridazine-2,5-dione (80.27 mg, 144.02 μmol, 76.6% yield, 100.0% purity) was obtained as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 10.81 (s, 1H), 7.44 (d, J = 1.8, 10.1 Hz, 1H), 7.35 (d, J = 8.6 Hz, 1H), 7.24 (d, J = 7.6 Hz, 1H), 6.84-6.70 (m, 3H), 6.53 (t, J = 8.5 Hz, 1H), 6.04 (td, J = 6.0, 10.5, 17.0 Hz, 1H), 5.37-5.24 (m, 2H), 4.85 (d, J = 4.9 Hz, 2H), 3.83 (s, 2H), 3.16 (s, 3H), 1.81 (s, 3H); ES-LCMS m/z 557.9 [M+H] ⁺ .

I-46

Step 1 : 4 - Methylbenzenesulfonate N - Methyl -1- (3- Nitrophenyl ) Methanesulfonamide

To a solution of (3-nitrophenyl)methanesulfonyl chloride (500 mg, 2.12 mmol, 1 eq) in THF (1 mL) was added DIEA (548.47 mg, 4.24 mmol, 739.17 μL, 2 eq) and MeNH _2.EtOH (197.69 mg, 6.37 mmol, 212.18 μL, 3 eq) was added. The mixture was stirred at 25°C for 1 hour. The mixture was quenched with water (30 mL) and extracted using DCM (30 mL x 3). The combined organic phases were washed with NaHCO ₃ (30 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 (PE/EtOAc = 100/1). From 1/1, purified by TLC: PE/EtOAc = 2/1, R _f = 0.4) to give 4-methylbenzenesulfonate N -methyl-1-(3-nitrophenyl)methanesulfonamide (200 mg, 861.19 μmol, 40.6% yield, 99.1% purity) was obtained as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.32-8.20 (m, 2H), 7.79 (d, J = 7.6 Hz, 1H), 7.64-7.58 (m, 1H), 4.35 (s, 2H), 4.27 -4.13 (m, 1H), 2.79 (d, J = 5.4 Hz, 3H).

Step 2 : 1 -(3- aminophenyl ) -N - methylmethanesulfonamide

Pd /C ₍ 200 mg, 861.19 μmol, 10.0%, 1 eq) was added. The suspension was degassed under vacuum and purged several times with H ₂ . The mixture was stirred at 25° C. under H ₂ (15 psi) for 1 hour. The reaction mixture was filtered and concentrated under reduced pressure to give 1-(3-aminophenyl)- N -methyl-methanesulfonamide (180 mg, 853.90 μmol, 99.2% yield, 95.0% purity) as a yellow solid, which was added It was used in the next step without purification. ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 7.22-7.17 (m, 1H), 7.17-7.14 (m, 1H), 6.75-6.73 (m, 2H), 6.71-6.67 (m, 2H), 6.66- 6.57 (m, 1H), 4.18 (s, 2H), 2.02 (s, 3H).

Step 3 : 1-(3-((3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-2,4,7-trioxo-1,2,3 ,4,7,8-hexahydropyrido[2,3-d]pyrimidin-5-yl)amino)phenyl)- N -methylmethanesulfonamide

2,6-lutidine (119.77 mg, 1.12 mmol) in a solution of 1-(3-aminophenyl)- N -methyl-methanesulfonamide (150 mg, 711.58 μmol, 95.0%, 1.91 eq) in DMA (3 mL) , 130.19 μL, 3 eq) and [3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2, 3- d ]pyrimidin-5-yl] 4-methylbenzenesulfonate (250 mg, 372.60 μmol, 90.0%, 1 eq) was added. The mixture was stirred at 140° C. under a microwave (2 bar) for 3 hours. The mixture was diluted with water (30 mL) and extracted using ethyl acetate (30 mL x 3). The organic layer was dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (HCl) - ACN]; B%: 46%-66%, 10 min) and lyophilized to obtain the product, which was added with saturated aqueous NaHCO ₃ to pH = 7 and purified using EtOAc (20 mL x 3). After extraction, the combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give 1-(3-((3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6 ,8-dimethyl-2,4,7-trioxo-1,2,3,4,7,8-hexahydropyrido [2,3- d ] pyrimidin-5-yl) amino) phenyl) - N -Methylmethanesulfonamide (45 mg, 58.55 μmol, 15.7% yield, 86.6% purity) was obtained as a yellow solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.85 (dd, J = 1.8, 9.5 Hz, 1H), 7.76 (d, J = 8.2 Hz, 1H), 7.37 (t, J = 7.8 Hz, 1H) , 7.22-7.14 (m, 2H), 7.08-6.99 (m, 2H), 4.33 (s, 2H), 2.95 (s, 3H), 2.82-2.76 (m, 1H), 2.69 (s, 3H), 1.67 (s, 3H), 1.16-1.09 (m, 2H), 0.81 (d, J = 2.6 Hz, 2H); ES-LCMS m/z 666.0 [M+H] ⁺ .

Step 4 : 1-(3-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo-3,4 ,6,7-Tetrahydropyrido[4,3- d ]pyrimidin-1(2H ) -yl)phenyl) -N -methylmethanesulfonamide

1-(3-((3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-2,4,7-trioxo-1 in THF (1 mL) 2,3,4,7,8-hexahydropyrido [2,3- d ] pyrimidin-5-yl) amino) phenyl) - N -methylmethanesulfonamide (45 mg, 58.15 μmol, 86.0%, 1 To the solution of eq), one drop of NaOMe/MeOH (20 wt%, 58.15 μmol, 1 eq) was added. The mixture was stirred at 25°C for 1 hour under N ₂ atmosphere. AcOH was added to the residue until pH = 7 and extracted using EtOAc (20 mL x 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um; mobile phase: [water (NH ₃ H ₂ O + NH ₄ HCO ₃ )-ACN]; B%: 53%-83%, 10 minutes) and lyophilized to obtain 1-(3-(3-cyclopropyl-5-((2-fluo Ro-4-iodophenyl) amino) -6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido [4,3- d ] pyrimidine-1 ( 2 H )-yl)phenyl)- N -methylmethanesulfonamide (10.01 mg, 14.96 μmol, 25.7% yield, 99.4% purity) was obtained as a white solid. ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 7.66 (dd, J = 1.7, 10.3 Hz, 1H), 7.57 (d, J = 8.3 Hz, 1H), 7.51-7.47 (m, 2H), 7.43 ( s, 1H), 7.39 (dd, J = 3.2, 5.6 Hz, 1H), 6.86 (t, J = 8.4 Hz, 1H), 4.37 (s, 2H), 3.20 (s, 3H), 2.76-2.69 (m , 1H), 2.67 (s, 3H), 1.34 (s, 3H), 1.08-1.03 (m, 2H), 0.81-0.74 (m, 2H); ES-LCMS m/z 666.1 [M+H] ⁺ .

I-33

Step 1 : 6 - Cyclopropyl -4- (2- Fluoro -4- Iodo - Anilino ) -3,7-dimethyl-8- [2-methyl-3- (methylsulfamoylamino) phenyl] pyrano [3,2-c]pyridine-2,5-dione

8-(3-Amino-2-methyl-phenyl)-6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-3,7-dimethyl in DCM (2 mL) at 0°C. -In a stirred solution of pyrano[3,2-c]pyridine-2,5-dione (50 mg, 70.01 μmol, 80% purity, 1 eq) and TEA (28.34 mg, 280.02 μmol, 38.98 μL, 4 eq) N -methylsulfamoyl chloride (18.14 mg, 140.01 μmol, not measurable purity, 2 eq) was added. The reaction mixture was stirred at 0° C. for 5 hours under N ₂ atmosphere. The reaction mixture was quenched by adding saturated aqueous NaHCO ₃ (30 mL) at 0° C. and extracted using DCM (30 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give a residue, which was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um). ; Mobile phase: purified with [water(NH ₃ H ₂ O + NH ₄ HCO ₃ )-ACN]; B%: 53%-83%, 10 minutes) and then lyophilized to produce 6-cyclopropyl-4-( 2-fluoro-4-iodo-anilino)-3,7-dimethyl-8-[2-methyl-3-(methylsulfamoylamino)phenyl]pyrano[3,2-c]pyridine-2, 5-Dione (6.22 mg, 9.36 μmol, 13.4% yield, 100.0% purity) was obtained as a white solid. ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 11.30 (s, 1H), 8.82 (s, 1H) _, 7.71 (d, J = 1.7, 10.3 Hz, 1H), 7.51 (d, J = 8.3 Hz) , 1H), 7.40 (d, J = 7.8 Hz, 1H), 7.27 (t, J = 7.7 Hz, 1H), 7.14 (q, J = 4.9 Hz, 1H), 7.01 (d, J = 7.6 Hz, 1H) ), 6.87 (t, J = 8.7 Hz, 1H), 3.03 (s, 1H), 2.55 (d, J = 5.1 Hz, 3H), 2.20 (s, 3H), 2.03 (s, 3H), 1.43 (s) , 3H), 1.23-1.17 (m, 2H), 0.96-0.84 (m, 2H); ES-LCMS m/z 665.0 [M+H] ⁺ .

I-47 & I-48

Step 1 : Methanesulfinyl chloride

_Sulfuryl chloride ( 21.79 g, 161.47 mmol, 16.14 mL, 3 eq) was added dropwise. The mixture was stirred at -20°C for 2 hours under N ₂ atmosphere. The mixture was slowly warmed to 20°C and stirred under N ₂ atmosphere at 20°C for 1 hour and at 35°C for 1 hour. The mixture was concentrated under reduced pressure to give methanesulfinyl chloride (10.6 g, crude) as a colorless oil, which was used in the next step without further purification. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 3.38 (s, 3H).

Step 2 : ( S ) -N- [3-[3- cyclopropyl -5-(2- fluoro -4- iodo - anilino )-6,8-dimethyl-2,4,7-trioxo- pyrido[4,3-d]pyrimidin-1-yl]phenyl]methanesulfinamide and ( R ) -N- [3-[3-cyclopropyl-5-(2-fluoro-4-iodo- Anilino)-6,8-dimethyl-2,4,7-trioxo-pyrido[4,3-d]pyrimidin-1-yl]phenyl]methanesulfinamide

1-(3-aminophenyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-pyrido[4,3-] in DCM (10 mL) d]pyrimidine-2,4,7-trione (200 mg, 348.82 μmol, 100% purity, 1 eq) and methanesulfinyl in Et ₃ N (363.50 mg, 3.59 mmol, 0.5 mL, 10.30 eq) Chloride (200 mg, 2.03 mmol, not measurable purity, 5.82 eq) was added. The mixture was stirred at 25°C for 0.5 hours. TLC (PE/EtOAc = 2/3, R _f = 0.15) showed that the starting material was almost consumed. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (PE/EtOAc = from 100/1 to 0/1, TLC: PE/EtOAc = 2/3, R _f = 0.15) N -[3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-2,4,7-trioxo-pyrido[4,3 -d]pyrimidin-1-yl]phenyl]methanesulfinamide (200 mg, 91.9% purity) was obtained as a white solid. The product (160 mg) was _{subjected to} preparative HPLC (column: Welch After purification, the desired compound was obtained as a white solid, which was purified by SFC (column: DAICEL CHIRALCEL OD-H (250 mm*30 mm, 5 um); mobile phase: [0.1% NH ₃ H ₂ O EtOH]; B%: 45 %-45%) to obtain peak 1 and peak 2. Peak 1 was concentrated under reduced pressure to obtain a residue, which _was subjected to preparative _HPLC (column: Welch %-73%, 11 min) and then lyophilized to give ( S ) -N- [3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6, 8-dimethyl-2,4,7-trioxo-pyrido[4,3-d]pyrimidin-1-yl]phenyl]methanesulfinamide (13.75 mg, 20.91 μmol, 9.0% yield, 96.6% purity, SFC : R _t = 4.554, ee = 100%, [α] ^27.8 _D = -36.364 (MeOH, c = 0.011 g/100 mL)) was obtained as a white solid. ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 7.66 (dd, J = 1.8, 10.1 Hz, 1H), 7.59-7.54 (m, 1H), 7.39 (t, J = 8.1 Hz, 1H), 7.17-7.08 (m, 2H), 7.05-7.00 (m, 1H), 6.86 (t, J = 8.5 Hz, 1H), 3.20 (s, 3H), 2.84 (s, 3H), 2.71 (tt, J = 3.8, 7.2 Hz, 1H), 1.40 (s, 3H), 1.10-1.00 (m, 2H), 0.79-0.70 (m, 2H); ES-LCMS m/z 636.1 [M+H] ⁺ . Peak 2 was concentrated under reduced pressure to obtain a residue, which was subjected _to preparative _HPLC (column: Welch -70%, 10 minutes) to obtain the desired compound as a white solid, which was purified by SFC (column: DAICEL CHIRALCEL OD-H (250 mm*30 mm, 5 um); mobile phase: [0.1% NH ₃ H ₂ O ETOH]; B%: 45%-45%, min) and then lyophilized to obtain the residue, which was subjected to preparative HPLC (column: Welch Xtimate C18 150*25 mm*5 um; mobile phase: [water] (NH ₄ HCO ₃ )-ACN]; B%: 40%-70%, 11 min) and then lyophilized to give ( R )- N -[3-[3-cyclopropyl-5-(2- Fluoro-4-iodo-anilino)-6,8-dimethyl-2,4,7-trioxo-pyrido[4,3-d]pyrimidin-1-yl]phenyl]methanesulfinamide (12.41 mg, 19.05 μmol, 8.2% yield, 97.6% purity, SFC: R _t = 4.991, ee = 89.18%, [α] ^27.7 _D = +107.692 (MeOH, c = 0.013 g/100 mL)) as a white solid. did. ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 7.66 (dd, J = 1.8, 10.2 Hz, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.39 (t, J = 8.0 Hz, 1H), 7.17-7.10 (m, 2H), 7.02 (dd , J = 1.1, 8.0 Hz, 1H), 6.86 (t, J = 8.5 Hz, 1H), 3.20 (s, 3H), 2.84 (s, 3H), 2.71 (tt, J = 3.7, 7.2 Hz, 1H) , 1.40 (s, 3H), 1.08-1.03 (m, 2H), 0.78-0.73 (m, 2H); ES-LCMS m/z 636.0 [M+H] ⁺ .

I-49

Step 1 : Ethyl 4- Chloro -3- (3- Nitrophenyl ) -6-oxo-1 H - Pyridazine -5- Carboxylate

To a solution of ethyl 4-hydroxy-3-(3-nitrophenyl)-6-oxo-1 H -pyridazine-5-carboxylate (2.5 g, 7.37 mmol, 1 eq) in DCM (30 mL) (COCl ) ₂ (935.60 mg, 7.37 mmol, 645.24 μL, 1 eq) was added. The mixture was stirred at 35°C for 3 hours. The reaction mixture was quenched by adding water (30 mL) and extracted using DCM (40 mL x 3). 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 to which MeOH (5 mL) was added and incubated at 25° C. for 2 h. It was stirred. The slurry was filtered and the cake was rinsed with PE (2 x 3 mL). The solid was collected and dried in vacuo to obtain ethyl 4-chloro-3-(3-nitrophenyl)-6-oxo-1H-pyridazine-5-carboxylate (1.9 g, 5.69 mmol, 77.2% yield, 97.0% purity). ) was obtained as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 11.67 (s, 1H), 8.48 (s, 1H), 8.38 (d, J = 8.1 Hz, 1H), 7.92 (d, J = 7.8 Hz, 1H), 7.74-7.66 (m, 1H), 4.51 (q, J = 7.1 Hz, 2H), 1.43 (t, J = 7.1 Hz, 3H); ES-LCMS m/z 324.0, 326.0 [M+H] ⁺ .

Step 2 : Ethyl 5- Chloro -2- Cyclopropyl -6- (3- Nitrophenyl ) -3-oxo- Pyridazine -4- Carboxylate

Ethyl 4-chloro-3-(3-nitrophenyl)-6-oxo-1H-pyridazine-5-carboxylate (1.4 g, 4.20 mmol, 1 eq), cyclopropylboronic acid (720.75) in DCE (20 mL) mg, 8.39 mmol, 2 eq), 2-(2-pyridyl)pyridine (1.64 g, 10.49 mmol, 2.5 eq), Na ₂ CO ₃ (533.60 mg, 5.03 mmol, 1.2 eq) and Cu(OAc) ₂ ( The mixture (914.43 mg, 5.03 mmol, 1.2 eq) was degassed, purged three times with N ₂ and the mixture was stirred at 70° C. for 24 hours under N ₂ atmosphere. The reaction mixture was diluted with water (100 mL) and extracted using DCM (60 mL x 3). The combined organic phases were washed with brine (50 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 (PE/EtOAc = 1/0 to 3). /1, TLC: PE/EtOAc = 3/1, R _f = 0.43) to give ethyl 5-chloro-2-cyclopropyl-6-(3-nitrophenyl)-3-oxo-pyridazine-4- The carboxylate (400 mg, 989.69 μmol, 23.6% yield, 90.0% purity) was obtained as a yellow gum. ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 8.42 (d, J = 1.8 Hz, 1H), 8.35 (td, J = 1.1, 8.3 Hz, 1H), 7.87 (d, J = 7.6 Hz, 1H), 7.70-7.65 (m, 1H), 4.51 (q, J = 7.1 Hz, 2H), 4.22 (t, J = 4.0, 7.6 Hz, 1H), 1.43 (t, J = 7.2 Hz, 3H), 1.22-1.16 (m, 2H), 1.13-1.06 (m, 2H); ES-LCMS m/z 364.0, 366.0 [M+H] ⁺ .

Step 3 : Ethyl 2- cyclopropyl -5-( methylamino )-6-(3- nitrophenyl )-3-oxo- pyridazine -4-carboxylate

of ethyl 5-chloro-2-cyclopropyl-6-(3-nitrophenyl)-3-oxo-pyridazine-4-carboxylate (400 mg, 989.69 μmol, 1 eq) in DCM (3 mL) at 0°C. MeNH ₂ (1.02 g, 9.90 mmol, 30.0% purity, 10 eq) was added to the solution. The mixture was stirred at 20°C for 2 hours. The solvent was removed to obtain ethyl 2-cyclopropyl-5-(methylamino)-6-(3-nitrophenyl)-3-oxo-pyridazine-4-carboxylate (300 mg, 669.74 μmol, 67.7% yield, 80.0% Purity) was obtained as a yellow solid. ^1H NMR (500 MHz, CDCl ₃ ) δ ppm 8.33-8.30 (m, 2H), 7.79 (d, J = 7.6 Hz, 1H), 7.70-7.66 (m, 1H), 4.43 (q, J = 7.1 Hz) , 2H), 4.02-3.95 (m, 1H), 2.69 (d, J = 5.3 Hz, 3H), 1.44 (t, J = 7.1 Hz, 2H), 1.46-1.42 (m, 1H), 1.03-0.94 ( m, 4H); ES-LCMS m/z 359.1 [M+H] ⁺ .

Step 4 : 2 - Cyclopropyl -5-( methylamino )-6-(3- nitrophenyl ) pyridazin -3-one

of ethyl 2-cyclopropyl-5-(methylamino)-6-(3-nitrophenyl)-3-oxo-pyridazine-4-carboxylate (300 mg, 669.74 μmol, 1 eq) in DMSO (5 mL) LiCl (283.93 mg, 6.70 mmol, 10 eq) was added to the solution. The mixture was stirred at 150°C for 8 hours. The reaction mixture was diluted with water (30 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 (PE/EtOAc = 1/0 to 0). /1, TLC: PE/EtOAc = 0/1, R _f = 0.15) to give 2-cyclopropyl-5-(methylamino)-6-(3-nitrophenyl)pyridazin-3-one (200 mg, 628.74 μmol, 93.9% yield, 90.0% purity) was obtained as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 8.38-8.27 (m, 2H), 7.83 (d, J = 7.6 Hz, 1H), 7.73-7.66 (m, 1H), 5.92 (s, 1H), 4.03 (t, J = 3.9, 7.5 Hz, 1H), 2.83 (d, J = 3.4 Hz, 3H), 1.09-0.93 (m, 4H); ES-LCMS m/z 287.1 [M+H] ⁺ .

Step 5 : 6 - Cyclopropyl -4- Hydroxy -1,3-dimethyl-8- (3-nitrophenyl) pyrido [2,3-d] pyridazine -2,5-dione

To a solution of 2-cyclopropyl-5-(methylamino)-6-(3-nitrophenyl)pyridazin-3-one (200 mg, 628.74 μmol, 1 eq) in Ac ₂ O (3 mL) was added 2-methyl. Propanedioic acid (222.74 mg, 1.89 mmol, 152.56 μL, 3 eq) was added. The mixture was stirred at 110°C for 2 hours. The solvent was removed to give a residue, which was purified by preparative TLC (PE/EtOAC = 3/1, TLC: PE/EtOAC = 3/1, R _f = 0.24) to give 6-cyclopropyl-4-hydroxy -1,3-dimethyl-8-(3-nitrophenyl)pyrido[2,3-d]pyridazine-2,5-dione (200 mg, 434.38 μmol, 69.1% yield, 80.0% purity) was obtained as a white solid. was obtained. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 12.80 (s, 1H), 8.37 (td, J = 2.2, 7.0 Hz, 1H), 8.33 (d, J = 1.5 Hz, 1H), 7.72-7.68 (m , 2H), 4.16-4.09 (m, 1H), 3.06 (s, 3H), 2.16 (s, 3H), 1.20-1.15 (m, 2H), 1.15-1.09 (m, 2H); ES-LCMS m/z 369.0 [M+H] ⁺ .

Step 6 : [6- Cyclopropyl -1,3-dimethyl-8- (3- nitrophenyl ) -2,5- dioxo - pyrido [2,3-d] pyridazin -4-yl] 4- methyl Benzenesulfonate

6-cyclopropyl-4-hydroxy-1,3-dimethyl-8-(3-nitrophenyl)pyrido[2,3-d]pyridazine-2,5-dione (200 mg) in DCM (8 mL) , 434.38 μmol, 1 eq), DIEA (561.40 mg, 4.34 mmol, 756.61 μL, 10 eq) and 4-methylbenzenesulfonyl chloride (414.06 mg, 2.17 mmol, 5 eq) were added. The mixture was stirred at 50°C for 12 hours. The solvent was removed to obtain a residue, which was purified by preparative TLC (PE/EtOAC = 3/1, TLC: PE/EtOAc = 3/1, R _f = 0.24) to give [6-cyclopropyl-1,3 -Dimethyl-8-(3-nitrophenyl)-2,5-dioxo-pyrido[2,3-d]pyridazin-4-yl] 4-methylbenzenesulfonate (200 mg, 371.27 μmol, 85.5% Yield, 97.0% purity) was obtained as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.38-8.30 (m, 2H), 7.98 (d, J = 8.3 Hz, 2H), 7.77-7.63 (m, 2H), 7.41 (d, J = 8.0 Hz) , 2H), 4.22-4.06 (m, 1H), 3.08 (s, 3H), 2.49 (s, 3H), 1.85 (s, 3H), 1.12-1.01 (m, 4H); ES-LCMS m/z 523.1 [M+H] ⁺ .

Step 7 : 6 - Cyclopropyl -4- (2- Fluoro -4- Iodo - Anilino ) -1,3-dimethyl-8- (3-nitrophenyl) pyrido [2,3-d] pyridazine -2,5-dione

To a solution of 2-fluoro-4-iodo-aniline (263.99 mg, 1.11 mmol, 3 eq) in THF (3 mL) was added NaH (89.10 mg, 2.23 mmol, 60.0% purity, 6 eq), After stirring at 0°C for 30 min, [6-cyclopropyl-1,3-dimethyl-8-(3-nitrophenyl)-2,5-dioxo-pyrido[2,3-d]pyri minced-4-yl] 4-methylbenzenesulfonate (200 mg, 371.27 μmol, 1 eq) was added. The reaction temperature was slowly warmed to 25°C and stirred at 25°C for 1 hour. The reaction mixture was quenched by adding water (20 mL) and extracted using EtOAc (30 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 analyzed by preparative TLC (PE/EtOAC = 1/1, TLC: PE/ EtOAc = 1/1, R _f = 0.35) to give 6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-8-(3-nitrophenyl) Pyrido[2,3-d]pyridazine-2,5-dione (100 mg, 156.64 μmol, 42.2% yield, 92.0% purity) was obtained as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 10.78 (s, 1H), 8.44-8.27 (m, 2H), 7.75-7.65 (m, 2H), 7.46 (d, J = 1.8, 10.0 Hz, 1H) , 7.37 (d, J = 8.5 Hz, 1H), 6.56 (t, J = 8.5 Hz, 1H), 4.10-4.01 (m, 1H), 3.07 (s, 3H), 1.82 (s, 3H), 1.19- 1.07 (m, 4H); ES-LCMS m/z 588.0 [M+H] ⁺ .

Step 8 : 8 -(3- aminophenyl )-6- cyclopropyl -4-(2- fluoro- 4- iodo - anilino )-1,3-dimethyl-pyrido[2,3-d]pyri Minced-2,5-dione

6-Cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-8- in EtOH (1.5 mL), H ₂ O (1.5 mL) and THF (1.5 mL). (3-nitrophenyl)pyrido[2,3-d]pyridazine-2,5-dione (100 mg, 156.64 μmol, 1 eq) was added to a solution of NH ₄ Cl (83.79 mg, 1.57 mmol, 10 eq) and Fe (43.74 mg, 783.19 μmol, 5 eq) was added. The mixture was stirred at 90°C for 3 hours. The reaction mixture was filtered to remove insoluble material. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um; mobile phase: [water (NH ₃ H ₂ O + NH ₄ HCO ₃ )-ACN]; B%: 64%-94%, 10 minutes) and then lyophilized to produce 8-(3-aminophenyl)-6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-pyrido. [2,3-d]pyridazine-2,5-dione (55.23 mg, 99.09 μmol, 63.3% yield, 100.0% purity) was obtained as a yellow solid. ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 7.55 (d, J = 1.8, 10.3 Hz, 1H), 7.45 (d, J = 8.5 Hz, 1H), 7.21 (t, J = 7.7 Hz, 1H), 6.82 (d, J = 1.5, 8.0 Hz, 1H), 6.79-6.76 (m, 1H), 6.71 (d, J = 7.7 Hz, 1H), 6.64 (t, J = 8.6 Hz, 1H), 4.04 (t , J = 3.9, 7.5 Hz, 1H), 3.13 (s, 3H), 1.76 (s, 3H), 1.17-1.11 (m, 2H), 1.07-0.99 (m, 2H); ES-LCMS m/z 558.0 [M+H] ⁺ .

I-50

Step 1 : N -[3-[6- Cyclopropyl -4- (2- Fluoro -4- Iodo - Anilino ) -1,3-dimethyl-2,5-dioxo-pyrido [2,3-d] Pyridazin-8-yl]phenyl]acetamide

8-(3-aminophenyl)-6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-pyrido[2,3-] in DCM (3 mL) d]pyridazine-2,5-dione (20 mg, 35.88 μmol, 1 eq) in a solution of acetyl chloride (5.63 mg, 71.77 μmol, 5.12 μL, 2 eq) and TEA (14.52 mg, 143.53 μmol, 19.98 μL; 4 eq) was added. The mixture was stirred at 25°C for 1 hour. The solvent was removed to obtain a residue, which was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um; mobile phase: [water (NH ₃ H ₂ O + NH ₄ HCO ₃ )-ACN]; B %: 55%-85%, 10 minutes) and then lyophilized to obtain N -[3-[6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3. -Dimethyl-2,5-dioxo-pyrido[2,3-d]pyridazin-8-yl]phenyl]acetamide (7.96 mg, 13.28 μmol, 37.0% yield, 100.0 purity) was obtained as a yellow solid. . ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 7.76 (s, 1H), 7.62 (d, J = 8.1 Hz, 1H), 7.55 (d, J = 1.7, 10.3 Hz, 1H), 7.49-7.40 ( m, 2H), 7.20 (d, J = 7.6 Hz, 1H), 6.64 (t, J = 8.6 Hz, 1H), 4.05 (t, J = 3.8, 7.5 Hz, 1H), 3.09 (s, 3H), 2.14 (s, 3H), 1.76 (s, 3H), 1.17-1.09 (m, 2H), 1.09-1.00 (m, 2H); ES-LCMS m/z 600.0 [M+H] ⁺ .

I-20

Step 1 : 6 - Cyclopropyl -4- (2- Fluoro -4- Iodo - Anilino ) -1,3-dimethyl-8- [3- (methylsulfamoylamino) phenyl] pyrido [2,3 -d]pyridazine-2,5-dione

8-(3-aminophenyl)-6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-pyrido[2,3-] in DCM (2 mL) d]pyridazine-2,5-dione (20 mg, 35.88 μmol, 1 eq) in a solution of TEA (3.63 mg, 35.88 μmol, 4.99 μL, 1 eq) and N -methylsulfamoyl chloride (9.30 mg, 71.77 μmol). , 2 eq) was added. The mixture was stirred at 0°C for 1 hour. The solvent was removed to obtain a residue, which was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm* 5 um; mobile phase: [water(NH ₃ H ₂ O + NH ₄ HCO ₃ )-ACN]; B %:51%-81%, 10 minutes) and then lyophilized to obtain 6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-8-[3 -(Methylsulfamoylamino)phenyl]pyrido[2,3-d]pyridazine-2,5-dione (7.62 mg, 11.46 μmol, 31.9% yield, 97.9% purity) was obtained as a yellow solid. ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 7.55 (d, J = 1.8, 10.3 Hz, 1H), 7.49-7.40 (m, 2H), 7.33-7.26 (m, 2H), 7.18 (d, J = 7.6 Hz, 1H), 6.65 (t, J = 8.6 Hz, 1H), 4.05 (t, J = 3.9, 7.5 Hz, 1H), 3.09 (s, 3H), 2.61 (s, 3H), 1.77 (s , 3H), 1.18-1.10 (m, 2H), 1.09-0.99 (m, 2H); ES-LCMS m/z 651.0 [M+H] ⁺ .

I-31

Step 1 : 5 -((1 H -indol-4-yl)amino)-3- cyclopropyl -1-(2- fluoro- 4- iodophenyl )-6,8-dimethylpyrido[2,3 - d ]pyrimidine-2,4,7(1 H , 3H ,8 H )-trione

[3-Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido in 1,4-dioxane (3 mL) [2,3- d ]pyrimidin-5-yl] 4-methylbenzenesulfonate (400 mg, 564.78 μmol, 90.0%, 1 eq), 1 H -indol-4-amine (111.96 mg, 847.17 μmol, 1.5 eq), a mixture of Xphos Pd G ₂ (24.30 mg _, 28.24 μmol, _0.05 eq) and It was stirred at 130°C for 6 hours. The mixture was diluted with water (30 mL) and extracted using ethyl acetate (50 mL x 3). The combined organic phases were concentrated in vacuo to give a residue, which was purified by flash silica gel chromatography (PE/EtOAc = 100/1 to 1/1, TLC: PE/EtOAc = 1/1, R _f = 0.26). 5-(( 1H -indol-4-yl)amino)-3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethylpyrido[2,3- d ]Pyrimidine-2,4,7( 1H , 3H , 8H )-trione (230 mg, 346.51 μmol, 61.4% yield, 90.0% purity) was obtained as a green solid. ^1H NMR (500 MHz, DMSO- d6 ) δ ppm 11.21 (s, 1H), 10.45 (s, 1H) _, 7.95 (dd, J = 1.8, 9.5 Hz, 1H), 7.74 (dd, J = 1.7, 8.4 Hz, 1H), 7.34 (t, J = 2.7 Hz, 1H), 7.28 (t, J = 8.1 Hz, 1H), 7.13 (d, J = 8.1 Hz, 1H), 7.02 (t, J = 7.8 Hz) , 1H), 6.46-6.38 (m, 2H), 2.75 (s, 3H), 2.70-2.65 (m, 1H), 2.07 (s, 3H), 1.02-0.98 (m, 2H), 0.69-0.64 (m , 2H); ES-LCMS m/z 598.0 [M+H] ⁺ .

Step 2 : 3 - Cyclopropyl -5-((2- fluoro- 4- iodophenyl )amino)-1-(1H - indol-4-yl)-6,8-dimethylpyrido[4,3 - d ]pyrimidine-2,4,7(1 H ,3 H ,6 H )-trione

5-((1 H -indol-4-yl)amino)-3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethylpyrido[2,3- d ] A solution of pyrimidine-2,4,7(1 H , 3H ,8 H )-trione (100 mg, 150.66 μmol, 90.0%, 1 eq) was added to NaOMe/MeOH (20 wt. %, 2 mL) in THF (2 mL). One drop (58.15 μmol, 1 eq) was added. The mixture was stirred at 25°C for 2 hours. AcOH was added to the residue until pH 7, and extracted using EtOAc (20 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 (PE/EtOAc = from 100/1 to 1/1, TLC: PE/EtOAc = 1/1, R _f = 0.3) to obtain 3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-1-(1 H -indol-4-yl)-6 , 8-dimethylpyrido [4,3- d ] pyrimidine-2,4,7 (1 H , 3 H , 6 H ) -trione (40 mg, 56.92 μmol, 37.8% yield, 85.0%) as green Obtained as a solid. ^1H NMR (400 MHz, DMSO- d ₆ ) δ ppm 11.29 (s, 1H), 11.12 (s, 1H), 7.84-7.73 (m, 1H), 7.55 (d, J = 8.6 Hz, 1H), 7.45 (d, J = 8.1 Hz, 1H), 7.38 (t, J = 2.6 Hz, 1H), 7.12 (t, J = 7.8 Hz, 1H), 7.01-6.88 (m, 2H), 6.22 (s, 1H) , 3.07 (s, 3H), 2.68-2.62 (m, 1H), 2.07 (s, 3H), 0.96-0.93 (m, 2H), 0.75-0.64 (m, 2H); ES-LCMS m/z 598.0 [M+H] ⁺ .

Step 3 : 1 -(1-Acetyl-1 H -indol-4-yl)-3- cyclopropyl -5-((2- fluoro- 4- iodophenyl )amino)-6,8-dimethylpyrido [4,3- d ]pyrimidine-2,4,7(1 H ,3 H ,6 H )-trione

3-Cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-1-(1 H -indol-4-yl)-6,8-dimethylpyrido[ 4,3- d ]pyrimidine-2,4,7(1 H ,3 H ,6 H )-trione (20 mg, 28.46 μmol, 85.0%, 1 eq) in Ac ₂ O (2.91 mg, 28.46 μmol, 2.67 μL, 1 eq) and DMAP (10.43 mg, 85.37 μmol, 3 eq) were added. The mixture was stirred at 0°C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (NH ₃ H ₂ O + NH ₄ HCO ₃ )-ACN] ; B%: 55%-85%, 10 minutes) and lyophilized to produce 1-(1-acetyl-1 H -indol-4-yl)-3-cyclopropyl-5-((2-fluoro- 4-iodophenyl) amino) -6,8-dimethylpyrido [4,3- d ] pyrimidine-2,4,7 (1 H , 3 H , 6 H ) -trione (9.66 mg, 15.03 μmol , 52.8% yield, 99.5%) was obtained as a white solid. ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 8.48 (d, J = 8.3 Hz, 1H), 7.79 (d, J = 3.9 Hz, 1H), 7.66 (d, J = 10.3 Hz, 1H), 7.58 (d, J = 8.3 Hz, 1H), 7.40 (t, J = 8.2 Hz, 1H), 7.22 (d, J = 7.6 Hz, 1H), 6.88 (t, J = 8.6 Hz, 1H), 6.55 (d , J = 3.9 Hz, 1H), 3.20 (s, 3H), 2.73 (s, 1H), 2.69 (s, 3H), 1.11 (s, 3H), 1.07 (d, J = 5.6 Hz, 2H), 0.80 (d, J = 3.9 Hz, 2H); ES-LCMS m/z 640.1 [M+H] ⁺ .

I-51 & I-52

Step 1 : ( S ) -N '-(3-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-tri oxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidin-1(2H ) -yl)phenyl)-N - methylmethanesulfonimideamide and ( R ) -N ' -(3-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo-3,4,6,7 -Tetrahydropyrido[4,3- d ]pyrimidin-1(2H ) -yl)phenyl) -N -methylmethanesulfonimideamide

N -[3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-2,4 in MeCN (20 mL) at 25°C under N ₂ atmosphere. ,7-trioxo-pyrido[4,3-d]pyrimidin-1-yl]phenyl]methanesulfinamide (100 mg, 135.81 μmol, 86.3% purity, 1 eq), MeNH ₂ (32 mg, 473.95 μmol , 100% purity, 3.49 eq, HCl) and t -BuOK (1 M, 1 mL, 7.36 eq) were added NCS (25 mg, 187.22 μmol, 1.38 eq). The mixture was stirred at 25°C for 12 hours under N ₂ atmosphere. The reaction mixture was diluted with H ₂ O (50 mL) and extracted using EtOAc (50 mL x 3). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue which was purified by flash silica gel chromatography (PE/EtOAc = from 100/1 to 0/1, TLC: PE/EtOAc = 1/2, R _f = 0.20). The desired fractions were concentrated under reduced pressure to obtain a residue, which was purified by SFC (column: (s,s) WHELK-O1 (250 mm*30 mm, 5 um); mobile phase: [0.1% NH ₃ H ₂ O/MeOH ]; B%: 60%-60%) 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 water (10 mL) and lyophilized to give ( S ) -N '-(3-(3-cyclopropyl-5-( (2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyri Mydin-1( 2H )-yl)phenyl) -N -methylmethanesulfonimideamide (5.32 mg, 8.01 μmol, 5.9% yield, 100.0% purity, SFC: R _t = 5.680, ee = 100%, [α] ^29.3 _D = -5.26 (MeOH, c = 0.114 g/100mL)). ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.67 (dd, J = 1.8, 10.1 Hz, 1H), 7.58 (d, J = 8.4 Hz, 1H), 7.30 (t, J = 8.0 Hz, 1H) , 7.14 (d, J = 7.8 Hz, 1H), 7.06 (s, 1H), 6.98 (d, J = 7.8 Hz, 1H), 6.87 (t, J = 8.5 Hz, 1H), 3.22 (s, 3H) , 3.12 (s, 3H), 2.74-2.72 (m, 1H), 2.70 (s, 3H), 1.43 (s, 3H), 1.10-1.05 (m, 2H), 0.79-0.75 (m, 2H); ES-LCMS m/z 665.1 [M+H] ⁺ . Peak 2 was concentrated under reduced pressure to give a residue, which was dissolved in MeCN (10 mL) and water (10 mL) and lyophilized to give ( R )- N '-(3-(3-cyclopropyl-5-( (2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyri Mydin-1( 2H )-yl)phenyl)- N -methylmethanesulfonimideamide (5.85 mg, 8.80 μmol, 6.5% yield, 100.0% purity, SFC: R _t = 6.804, ee = 100%, [α ] ^29.4 _D = +18.28 (MeOH, c = 0.186 g/100 mL)) was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.67 (dd, J = 1.8, 10.1 Hz, 1H), 7.58 (d, J = 8.4 Hz, 1H), 7.33-7.28 (m, 1H), 7.14 ( d, J = 7.8 Hz, 1H), 7.06 (s, 1H), 6.98 (d, J = 7.8 Hz, 1H), 6.87 (t, J = 8.5 Hz, 1H), 3.22 (s, 3H), 3.12 ( s, 3H), 2.75-2.72 (m, 1H), 2.70 (s, 3H), 1.43 (s, 3H), 1.10-1.05 (m, 2H), 0.79-0.75 (m, 2H); ES-LCMS m/z 665.0 [M+H] ⁺ .

I-54

Step 1 : 3-Cyclopropyl-1-(1,1-dioxido-3-oxo-2,3-dihydrobenzo[ d ]isothiazol-6-yl)-5-((2-fluoro -4-iodophenyl) amino) -6,8-dimethylpyrido [4,3-d] pyrimidine-2,4,7 (1 H , 3 H , 6 H ) -trione

[3-Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2,3-] in DMF (3 mL) d ]pyrimidin-5-yl] Cs ₂ CO ₃ (200.37 mg, 614.98 μmol, 2.5 eq) and 6-amino- 1,1-dioxo-1,2-benzothiazol-3-one (121.89 mg, 614.98 μmol, 2 eq) was added. The mixture was stirred at 130°C for 6 hours. The mixture was purified by preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (HCl)-ACN]; B%: 44%-64%, 10 min) and lyophilized for 3 -Cyclopropyl-1-(1,1-dioxido-3-oxo-2,3-hydrobenzo[ d ]isothiazol-6-yl)-5-((2-fluoro-4-iodo Phenyl) amino) -6,8-dimethylpyrido [4,3- d ] pyrimidine-2,4,7 (1 H , 3 H , 6 H ) -trione (6.74 mg, 9.69 μmol, 3.15% yield , 95.4% purity) was obtained as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 11.42 (s, 1H), 8.00 (s, 1H), 7.67 (s, 1H), 7.60 (s, 2H), 7.44-7.35 (m, 2H), 7.01 (s, 1H), 3.41 (s, 3H), 2.84-2.75 (m, 1H), 1.53 (s, 3H), 1.31-1.12 (m, 2H), 0.91-0.79 (m, 2H); ES-LCMS m/z 664.0 [M+H] ⁺ .

I-22

Step 1 : 1 -(5- aminopyridin -3-yl)-3- cyclopropyl -5-((2- fluoro- 4- iodophenyl )amino)-6,8-dimethylpyrido[4,3 - d ]pyrimidine-2,4,7(1 H ,3 H ,6 H )-trione

[3-Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2,3-] in DMF (5 mL) d ]pyrimidin-5-yl] pyridine-3,5-diamine (50.33 mg, 461.24 μmol, 1.2 eq) and 2 in a solution of 4-methylbenzenesulfonate (250 mg, 384.36 μmol, 98.0%, 1 eq). ,6-lutidine (102.96 mg, 960.91 μmol, 111.91 μL, 2.5 eq) was added. The mixture was stirred at 140°C for 6 hours under N ₂ atmosphere. The reaction mixture was diluted with H ₂ O (20 mL), extracted using DCM:IPA (3:1, 30 ml), and the organic layer was concentrated under reduced pressure to give 1-(5-aminopyridin-3-yl). -3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethylpyrido[4,3- d ]pyrimidine-2,4,7( 1H , 3 H , 6 H )-Trione (150 mg, 261.17 μmol, 68.0% yield, 100.0% purity) was obtained as a yellow solid. ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 8.08-7.93 (m, 1H), 7.78 (d, J = _8.6 Hz, 1H), 7.26 (d, J = 18.1 Hz, 2H), 7.19-7.09 (m, 1H), 6.85 (s, 1H), 6.51 (s, 1H), 6.48 (s, 1H), 4.89 (s, 1H), 2.83 (s, 3H), 2.37-2.25 (m, 1H), 1.79 (s, 3H), 0.96 (s, 2H), 0.57 (d, J = 14.7 Hz, 2H); ES-LCMS m/z 575.1 [M+H] ⁺ .

Step 2 : 3 - Cyclopropyl -5- (2- Fluoro -4- Iodo - Anilino ) -6,8-dimethyl-1- [5- (methylsulfamoylamino) -3-pyridyl] pyrido [4,3- d ]pyrimidine-2,4,7-trione

1-(5-Aminopyridin-3-yl)-3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethylpyrido[ In a solution of 4,3- d ]pyrimidine-2,4,7(1 H ,3 H ,6 H )-trione (95 mg, 165.41 μmol, 100.0%, 1 eq) was added N -methylsulfamoyl chloride ( 32.15 mg, 248.11 μmol, not determined, 1.5 eq) and TEA (50.21 mg, 496.22 μmol, 69.07 μL, 3 eq) were added. The mixture was stirred at 0°C for 3 hours under N ₂ atmosphere. The reaction mixture was diluted with H ₂ O (20 mL), extracted using DCM (30 mL 150*30 mm*5 um; Mobile phase: [Water (NH ₃ H ₂ O + NH ₄ HCO ₃ )-ACN]; B%: 20%-50%, 10 minutes) purified and lyophilized in 3-cycle Propyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-1-[5-(methylsulfamoylamino)-3-pyridyl]pyrido[4,3- d ]Pyrimidine-2,4,7-trione (16.52 mg, 24.67 μmol, 14.9% yield, 99.7% purity) was obtained as a white solid. ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 7.99 (ddd, J = 1.6, 4.9, _9.3 Hz, 1H), 7.76 (d, J = 8.3 Hz, 1H), 7.38-7.26 (m, 2H) , 7.18-7.04 (m, 2H), 6.05 (d, J = 15.5 Hz, 2H), 5.47 (s, 1H), 2.82 (d, J = 2.1 Hz, 3H), 2.39-2.35 (m, 3H), 2.33 (d, J = 1.8 Hz, 1H), 1.77 (d, J = 1.9 Hz, 3H), 1.00-0.91 (m, 2H), 0.58 (s, 2H); ES-LCMS m/z 668.0 [M+H] ⁺ .

I-41 and I-42

Step 1 : tert -Butyl N -[2-( chlorosulfonylamino )ethyl] carbamate

Sulfuryl chloride (850 mg, 6.30 mmol, 629.63 μL, 1.01 eq) and Et ₃ N (1.89 g, 18.73 mmol, 2.61 mL, 3 eq) in DCM (50 mL) at -40 to -50°C under N ₂ atmosphere. To the solution of tert -butyl N -(2-aminoethyl)carbamate (1 g, 6.24 mmol, 980.39 μL, 1 eq) in DCM (5 mL) was added dropwise. The mixture was stirred for 5 minutes at -40 to -50°C under N ₂ atmosphere to obtain tert -butyl N -[2-(chlorosulfonylamino)ethyl]carbamate (75 g, crude) as a colorless liquid. , which was used in the next step without further purification.

Step 2 : tert -Butyl N- [2-[[3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-2,4,7- trioxo-pyrido[4,3-d]pyrimidin-1-yl]phenyl]sulfamoylamino]ethyl]carbamate

1-(3-aminophenyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-pyrido[4,3-] in DCM (20 mL) d]pyrimidine-2,4,7-trione (90 mg, 156.97 μmol, 100% purity, 1 eq) and tert -butyl in a solution of Et ₃ N (145.40 mg, 1.44 mmol, 0.2 mL, 9.15 eq). N -[2-(chlorosulfonylamino)ethyl]carbamate (10 g, 38.65 mmol, not measurable purity, 246.23 eq) was added. The mixture was stirred at 15°C for 10 minutes. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (PE/EtOAc = from 100/1 to 1/1, TLC: PE/EtOAc = 3/4, R _f = 0.48) The desired compound (100 mg) was obtained. 50 mg of the desired compound (100 mg) was purified by preparative HPLC (column: Welch 10 minutes) and lyophilized to form tert -butyl N -[2-[[3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl- 2,4,7-trioxo-pyrido[4,3-d]pyrimidin-1-yl]phenyl]sulfamoylamino]ethyl]carbamate (16.07 mg, 20.07 μmol, 12.8% yield, 99.4% purity) was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.58 (dd, J = 1.8, 10.1 Hz, 1H), 7.49 (d, J = 8.2 Hz, 1H), 7.31 (t, J = 8.1 Hz, 1H), 7.17 (s, 1H), 7.13 (dd, J = 1.4, 8.1 Hz, 1H), 7.00-6.95 (m, 1H), 6.78 (t, J = 8.5 Hz, 1H), 3.13 (s, 3H), 3.03-2.98 (m, 2H), 2.97-2.92 ( m, 2H), 2.64 (tt, J = 3.8, 7.2 Hz, 1H), 1.33 (s, 9H), 1.32 (s, 3H), 1.02-0.96 (m, 2H), 0.71-0.66 (m, 2H) ; ES-LCMS m/z 796.1 [M+H] ⁺ .

Step 3 : 1 -[3-(2- Aminoethylsulfamoylamino )phenyl]-3- cyclopropyl -5-(2- fluoro- 4-iodo-anilino)-6,8-dimethyl-pyrido [4,3-d]pyrimidine-2,4,7-trione

tert -Butyl N- [2-[[3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-2,4 in DCM (2 mL) TFA in a solution of 7-trioxo-pyrido[4,3-d]pyrimidin-1-yl]phenyl]sulfamoylamino]ethyl]carbamate (50 mg, 62.84 μmol, not measurable purity, 1 eq) (3.08 g, 27.01 mmol, 2 mL, 429.83 eq) was added. The mixture was stirred at 20°C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um; mobile phase: [water (0.05% NH ₃ H ₂ O + 10 mM NH ₄ HCO) ₃ )-ACN]; B%: 45%-75%, 10 minutes) and lyophilized to obtain 1-[3-(2-aminoethylsulfamoylamino)phenyl]-3-cyclopropyl-5-(2 -Fluoro-4-iodo-anilino)-6,8-dimethyl-pyrido [4,3-d] pyrimidine-2,4,7-trione (16.78 mg, 23.94 umol, 38.09% yield, 99.223% purity) was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.68 (dd, J = 1.8, 10.1 Hz, 1H), 7.59 (d, J = 8.4 Hz, 1H), 7.41 (t, J = 8.0 Hz, 1H), 7.28-7.25 (m, 1H), 7.25-7.22 (m, 1H), 7.08-7.04 (m, 1H), 6.88 (t, J = 8.5 Hz, 1H), 3.22 (s, 3H), 3.06 (t, J = 6.1 Hz, 2H), 2.76-2.70 ( m, 3H), 1.41 (s, 3H), 1.10-1.05 (m, 2H), 0.80-0.76 (m, 2H); ES-LCMS m/z 696.0 [M+H] ⁺ .

I-47 and I-48

Step 1 : Methanesulfinyl chloride

_Sulfuryl chloride ( 21.79 g, 161.47 mmol, 16.14 mL, 3 eq) was added dropwise. The mixture was stirred at -20°C for 2 hours under N ₂ atmosphere. The mixture was slowly warmed to 20°C and stirred under N ₂ atmosphere at 20°C for 1 hour and at 35°C for 1 hour. The mixture was concentrated under reduced pressure to afford methanesulfinyl chloride (10.6 g, crude) as a colorless oil, which was used in the next step without further purification. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 3.38 (s, 3H).

Step 2 : ( S ) -N- [3-[3- cyclopropyl -5-(2- fluoro -4- iodo - anilino )-6,8-dimethyl-2,4,7-trioxo- pyrido[4,3-d]pyrimidin-1-yl]phenyl]methanesulfinamide and ( R ) -N- [3-[3-cyclopropyl-5-(2-fluoro-4-iodo- Anilino)-6,8-dimethyl-2,4,7-trioxo-pyrido[4,3-d]pyrimidin-1-yl]phenyl]methanesulfinamide

1-(3-aminophenyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-pyrido[4,3-] in DCM (10 mL) d]pyrimidine-2,4,7-trione (200 mg, 348.82 μmol, 100% purity, 1 eq) and methanesulfinyl in Et ₃ N (363.50 mg, 3.59 mmol, 0.5 mL, 10.30 eq) Chloride (200 mg, 2.03 mmol, not measurable purity, 5.82 eq) was added. The mixture was stirred at 25°C for 0.5 hours. TLC (PE/EtOAc = 2/3, R _f = 0.15) showed that the starting material was almost consumed. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (PE/EtOAc = from 100/1 to 0/1, TLC: PE/EtOAc = 2/3, R _f = 0.15) N -[3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-2,4,7-trioxo-pyrido[4,3 -d]pyrimidin-1-yl]phenyl]methanesulfinamide (200 mg, 91.9% purity) was obtained as a white solid. The product (160 mg) was _{subjected to} preparative HPLC (column: Welch After purification, the desired compound was obtained as a white solid, which was purified by SFC (column: DAICEL CHIRALCEL OD-H (250 mm*30 mm, 5 um); mobile phase: [0.1% NH ₃ H ₂ O EtOH]; B%: 45 %-45%)) was further separated to obtain peak 1 and peak 2. Peak 1 was concentrated under reduced pressure to obtain a residue, which _was subjected to preparative _HPLC (column: Welch %-73%, 11 min) and then lyophilized to give ( S ) -N- [3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6, 8-dimethyl-2,4,7-trioxo-pyrido[4,3-d]pyrimidin-1-yl]phenyl]methanesulfinamide (13.75 mg, 20.91 μmol, 9.0% yield, 96.6% purity, SFC : R _t = 4.554, ee = 100%, [α] ^27.8 _D = -36.364 (MeOH, c = 0.011 g/100 mL)) was obtained as a white solid. ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 7.66 (dd, J = 1.8, 10.1 Hz, 1H), 7.59-7.54 (m, 1H), 7.39 (t, J = 8.1 Hz, 1H), 7.17-7.08 (m, 2H), 7.05-7.00 (m, 1H), 6.86 (t, J = 8.5 Hz, 1H), 3.20 (s, 3H), 2.84 (s, 3H), 2.71 (tt, J = 3.8, 7.2 Hz, 1H), 1.40 (s, 3H), 1.10-1.00 (m, 2H), 0.79-0.70 (m, 2H); ES-LCMS m/z 636.1 [M+H] ⁺ . Peak 2 was concentrated under reduced pressure to obtain a residue, which was subjected _to preparative _HPLC (column: Welch -70%, 10 minutes) to obtain the desired compound as a white solid, which was purified by SFC (column: DAICEL CHIRALCEL OD-H (250 mm*30 mm, 5 um); mobile phase: [0.1% NH ₃ H ₂ O ETOH]; B%: 45%-45%, min), and then lyophilized to obtain a residue, which was subjected to preparative HPLC (column: Welch Xtimate C18 150*25 mm*5 um; mobile phase). :[Water(NH ₄ HCO ₃ )-ACN]; B%: 40%-70%, 11 minutes) and then lyophilized to produce ( R )- N -[3-[3-cyclopropyl-5- (2-Fluoro-4-iodo-anilino)-6,8-dimethyl-2,4,7-trioxo-pyrido[4,3-d]pyrimidin-1-yl]phenyl]methanesulfine The amide (12.41 mg, 19.05 μmol, 8.2% yield, 97.6% purity, SFC: R _t = 4.991, ee = 89.18%, [α] ^27.7 _D = +107.692 (MeOH, c = 0.013 g/100 mL)) was purified as white Obtained as a solid. ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 7.66 (dd, J = 1.8, 10.2 Hz, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.39 (t, J = 8.0 Hz, 1H), 7.17-7.10 (m, 2H), 7.02 (dd , J = 1.1, 8.0 Hz, 1H), 6.86 (t, J = 8.5 Hz, 1H), 3.20 (s, 3H), 2.84 (s, 3H), 2.71 (tt, J = 3.7, 7.2 Hz, 1H) , 1.40 (s, 3H), 1.08-1.03 (m, 2H), 0.78-0.73 (m, 2H); ES-LCMS m/z 636.0 [M+H] ⁺ .

I-55

Step 1 : 3 - Cyclopropyl -5- (2- Fluoro -4- Iodo - Anilino ) -6,8-dimethyl-1- [3- (methylamino) phenyl] pyrido [4,3-d ]Pyrimidine-2,4,7-trione

1-(3-aminophenyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-pyrido[4,3-] in THF (5 mL) To a solution of d]pyrimidine-2,4,7-trione (100 mg, 174.41 μmol, 100% purity, 1 eq) was added NaH (40 mg, 1.00 mmol, 60% purity, 5.73 eq). The mixture was stirred at 25°C for 0.5 hours. MeI (30 mg, 211.36 μmol, 13.16 μL, 1.21 eq) was added. The mixture was stirred at 25°C for 12 hours. The reaction mixture was quenched with H ₂ O (20 mL) and extracted using EtOAc (20 mL x 3). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was subjected to preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (HCl)] -ACN]; B%: 49%-69%, 10 minutes) and lyophilized to give 3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl- 1-[3-(methylamino)phenyl]pyrido[4,3-d]pyrimidine-2,4,7-trione (6 mg, 9.87 μmol, 5.7% yield, 96.6% purity) was purified as an off-white solid. Obtained. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.68 (dd, J = 1.8, 10.2 Hz, 1H), 7.66-7.62 (m, 1H), 7.60 (d, J = 8.4 Hz, 1H), 7.50 (t, J = 1.9 Hz, 1H), 7.47-7.40 (m, 2H), 6.90 (t, J = 8.5 Hz, 1H), 3.22 (s, 3H), 3.10 (s, 3H), 2.74 (tt, J = 3.7, 7.2 Hz, 1H), 1.37 (s, 3H), 1.11-1.06 (m, 2H), 0.81-0.76 (m, 2H); ES-LCMS m/z 588.1 [M+H] ⁺

Step 2 : 3 - cyclopropyl -5-(2- fluoro- 4 - iodo - anilino )-6,8-dimethyl-1-[3-[methyl(methylsulfamoyl)amino]phenyl]-2, 4,7-trioxo-pyrido[4,3-d]pyrimidine

3-Cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-1-[3-(methylamino)phenyl]pyrido in DCM (5 mL) at 0°C. [4,3-d]pyrimidine-2,4,7-trione (5 mg, 8.22 μmol, 96.6% purity, 1 eq) and DIEA (22.26 mg, 172.23 μmol, 30 μL, 20.95 eq). N -methylsulfamoyl chloride (10 mg, 77.18 μmol, 9.39 eq) was added. The mixture was stirred at 0°C for 10 minutes. The reaction mixture was diluted with H ₂ O (20 mL) and extracted using DCM (30 mL x 3). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was subjected to preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (HCl)] -ACN]; B%: 54%-74%, 10 minutes) and lyophilized to produce 3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl- 1-[3-[methyl(methylsulfamoyl)amino]phenyl]-2,4,7-trioxo-pyrido[4,3-d]pyrimidine (1.58 mg, 2.28 μmol, 27.8% yield, 98.3% Purity) was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 11.24 (s, 1H), 7.68 (dd, J = 1.3, 10.3 Hz, 1H), 7.58 (d, J = 8.4 Hz, 1H), 7.53-7.48 (m, 2H), 7.48-7.44 (m, 1H) , 7.34-7.25 (m, 1H), 6.88 (dt, J = 2.5, 8.5 Hz, 1H), 3.27 (s, 3H), 3.22 (d, J = 2.9 Hz, 3H), 2.73 (tt, J = 3.7) , 7.2 Hz, 1H), 2.65 (s, 3H), 1.39 (s, 3H), 1.13-1.02 (m, 2H), 0.83-0.75 (m, 2H); ES-LCMS m/z 680.9 [M+H] ⁺ .

I-82

Step 1 : tert -Butyl 5-(4- aminoanilino )-3- cyclopropyl -1-(2- fluoro- 4- iodo -phenyl)-6,8-dimethyl-pyrido[2,3- d ]pyrimidine-2,4,7-trione

[3-Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2,3-] in DMF (2 mL) d ]pyrimidin-5-yl]2,6-lutidine (155.17 mg, 1.45 mmol, 168.66 μL, 3 eq) in a solution of trifluoromethanesulfonate (300 mg, 482.70 μmol, 99.0%, 1 eq) and benzene-1,4-diamine (78.30 mg, 724.05 μmol, 1.5 eq) were added. The mixture was stirred at 60°C for 2 hours. The reaction mixture was quenched by addition of H ₂ O (20 mL) and extracted using EtOAc (30 mL x 3). 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 analyzed by preparative TLC (PE/EtOAc = 2/1, TLC: PE /EtOAc = 2/1, R _f = 0.20) to give tert -butyl 5-(4-aminoanilino)-3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6. ,8-Dimethyl-pyrido[2,3- d ]pyrimidine-2,4,7-trione (200 mg, 327.89 μmol, 67.9% yield, 94.0% purity) was obtained as a brown solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 10.16-10.03 (m, 1H), 8.04-7.87 (m, 1H), 7.77-7.60 (m, 1H), 7.32-7.13 (m, 1H), 6.85-6.65 (m, 2H), 6.63-6.45 (m, 2H), 5.24-4.99 (m, 2H), 2.73-2.67 (m, 3H), 1.51-1.34 (m, 3H), 1.09-0.89 (m) , 2H), 0.76-0.56 (m, 2H); ES-LCMS m/z 574.1 [M+H] ⁺ .

Step 2 : tert -Butyl 1-(4- aminophenyl )-3- cyclopropyl -5-(2- fluoro- 4- iodo -anilino)-6,8-dimethyl-pyrido[4,3- d ]pyrimidine-2,4,7-trione

5-(4-Aminoanilino)-3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-pyrido[2,3-] in THF (0.3 mL) d ] A mixture of pyrimidine-2,4,7-trione (100 mg, 163.95 μmol, 1 eq) and NaOMe/MeOH (163.95 μmol, 11 μL, 30.0%, 1 eq) was degassed and purified with N ₂ . After purging for 1 hour, the mixture was stirred at 25° C. for 1 hour under N ₂ atmosphere. The reaction mixture was quenched by adding AcOH (0.1 mL), then diluted with H ₂ O (20 mL) and extracted using EtOAc (30 mL x 3). 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 analyzed by preparative TLC (DCM/MeOH = 10/1, TLC: DCM). /MeOH=10/1, R _f = 0.80) to give tert -butyl 1-(4-aminophenyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6. ,8-Dimethyl-pyrido[4,3- d ]pyrimidine-2,4,7-trione (90 mg, 67.50 μmol, 41.2% yield, 43.0% purity) was obtained as a yellow solid. ES-LCMS m/z 574.0 [M+H] ⁺ .

Step 3 : tert -Butyl 3- cyclopropyl -5-(2- fluoro- 4 - iodo - anilino )-6,8-dimethyl-1-[4-(methylsulfamoylamino)phenyl]pyrido[ 4,3- d ]pyrimidine-2,4,7-trione

1-(4-aminophenyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-pyrido[4,3-] in DCM (1 mL) d ]pyrimidine-2,4,7-trione (43.00 mg, 75.00 μmol, 1 eq) in a solution of TEA (15.18 mg, 149.99 μmol, 20.88 μL, 2 eq) and N -methylsulfamoyl chloride (9.72 mg). , 75.00 μmol, 1 eq) was added. The mixture was stirred at 0°C for 1 hour. The reaction mixture was quenched by addition of H ₂ O (20 mL) and extracted using EtOAc (30 mL x 3). 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 subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um; mobile phase: [water (FA)-ACN]; B%: 56%-76%, 12 min) and then lyophilized to give tert -butyl 3-cyclopropyl-5-(2-fluoro- 4-iodo-anilino)-6,8-dimethyl-1-[4-(methylsulfamoylamino)phenyl]pyrido[4,3- d ]pyrimidine-2,4,7-trione (8.77 mg, 13.16 μmol, 17.6% yield, 100.0% purity) was obtained as a white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 11.23-10.93 (m, 1H), 10.00-9.81 (m, 1H), 7.90-7.70 (m, 1H), 7.59-7.52 (m, 1H), 7.50-7.44 (m, 1H), 7.29-7.23 (m, 2H), 7.22-7.15 (m, 2H), 6.99-6.82 (m, 1H), 3.12-3.01 (m, 3H), 2.63-2.57 (m) , 1H), 2.47-2.44 (m, 3H), 1.22 (s, 3H), 0.99-0.91 (m, 2H), 0.72-0.57 (m, 2H); ES-LCMS m/z 667.0 [M+H] ⁺ .

I-83

Step 1 : 3 - Cyclopropyl -1- (2- Fluoro -4- Iodophenyl ) -6,8-dimethyl-2,4,7- Trioxo -1,2,3,4,7,8- Hexahydropyrido[2,3- d ]pyrimidin-5-yl trifluoromethanesulfonate

3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-5-hydroxy-6,8-dimethyl-pyrido[2, 3- d ]trifluoride in a solution of pyrimidine-2,4,7-trione (3.2 g, 6.62 mmol, 1 eq) and 2,6-lutidine (1.21 g, 11.26 mmol, 1.31 mL, 1.7 eq). Romethylsulfonyl trifluoromethanesulfonate (3.74 g, 13.24 mmol, 2.19 mL, 2 eq) was added. Then, the mixture was stirred at 25°C for 12 hours under N ₂ atmosphere. To the reaction mixture was added successively saturated aqueous NaHCO ₃ (40 mL x 3), 1N HCl (40 mL x 3) and extracted using DCM (50 mL x 3). The _organic layer was washed with _brine (50 mL From 8/1, purified by TLC: PE/EtOAc = 1/1, R _f = 0.3) to give 3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-2 , 4,7-trioxo-1,2,3,4,7,8-hexahydropyrido [2,3- d ] pyrimidin-5-yl trifluoromethanesulfonate (4 g, 6.43 mmol, 97.1% yield, 98.9% purity) was obtained as a yellow solid. ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 7.95 (dd, J = 1.8, 9.4 Hz, 1H), 7.72 (dd, J = 1.3, 8.4 Hz, 1H), 7.30 (t, J = 8.1 Hz ₎ , 1H), 2.76 (s, 3H), 2.71-2.65 (m, 1H), 2.01 (s, 3H), 1.08-0.98 (m, 2H), 0.68-0.58 (m, 2H); ES-LCMS m/z 615.9 [M+H] ⁺ .

Step 2 : Methyl 2- chlorosulfonyl -5-nitro- benzoate

To a solution of methyl 2-amino-5-nitro-benzoate (5 g, 25.49 mmol, 1 eq) in HCl (12 M, 80 mL, 37.66 eq) at 0° C. was added NaNO ₂ (5 mL) in H ₂ O (5 mL). A solution of 1.76 g, 25.49 mmol, 1 eq) was added dropwise. The mixture was stirred at 0°C for 15 min and added to a solution of NaHSO ₃ (23.9 g, 229.67 mmol, 9.01 eq) in H ₂ O (50 mL) at 0°C followed by CuSO ₄ (406.83 mg, 2.55 mmol, 0.1 eq). ) was added, and the mixture was stirred at 0°C for 45 minutes. The mixture was diluted with water (100 mL) and extracted using EtOAc (80 mL x 3). The combined organic phases were washed with brine (100 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 (PE/EtOAc = 100/1 to From 5/1, purified by TLC: PE/EtOAc = 3/1, R _f = 0.51) to give methyl 2-chlorosulfonyl-5-nitro-benzoate (5.6 g, 18.02 mmol, 70.7% yield, 90.0% purity). ) was obtained as a yellow solid. ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 8.58 (d, J = 2.3 Hz, 1H), 8.54 (d, J = 2.3, 8.7 Hz, 1H), 8.40 (d, J = 8.7 Hz, 1H), 4.06 (s, 3H).

Step 3 : 5 -nitro-1,1- dioxo -1,2- benzothiazol -3-one

NH ₃ ^. A mixture of methyl 2-chlorosulfonyl-5-nitro-benzoate (5.6 g, 18.02 mmol, 90.0%, 1 eq) in H ₂ O (54.60 g, 436.23 mmol, 60 mL, 28.0% purity, 24.21 eq) It was stirred at 25°C for 16 hours. TLC (PE/EtOAc = 1/1, R _f = 0.04) showed that the starting material was completely consumed and one new spot was formed. The mixture was adjusted to pH 2 with HCl (12 M), diluted with water (150 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 5-nitro-1,1-dioxo-1,2-benzothiazol-3-one. (3.7 g, 14.59 mmol, 80.9% yield, 90.0% purity) was obtained as a yellow solid. ^1H NMR (500 MHz, CDCl ₃ ) δ ppm 8.88 (d, J = 2.0 Hz, 1H), 8.76 (d, J = 2.0, 8.4 Hz, 1H), 8.15 (d, J = 8.4 Hz, 1H).

Step 4 : 5 -Amino-1,1- dioxo -1,2- benzothiazol -3-one

5-nitro-1,1-dioxo-1,2-benzothiazole-3- in a solution of Pd/C (3 g, 14.59 mmol, 10% purity, 1 eq) in THF (50 mL) under N ₂ (3.7 g, 14.59 mmol, 90.0%, 1 eq) was added. The suspension was degassed under vacuum and purged several times with H ₂ . The mixture was stirred at 25° C. under H ₂ (15 psi) for 12 hours. The reaction mixture was filtered and the filter was concentrated to give a residue, to which EtOAc/MeOH (10/1, 10 mL) was added and stirred at 25°C for 2 hours. The slurry was filtered and the cake was rinsed with EtOAc (2 x 3 mL). The solid was collected and dried in vacuo to give 5-amino-1,1-dioxo-1,2-benzothiazol-3-one (2.8 g, 13.42 mmol, 91.9% yield, 95.0% purity) as a yellow solid. Obtained. ¹ H NMR (500 MHz, DMSO- d ₆ ) δ ppm 7.67 (d, J = 8.4 Hz, 1H), 7.02-6.90 (m, 2H).

Step 5 : 3 - Cyclopropyl -1-(2- fluoro- 4- iodo -phenyl)-6,8-dimethyl-5-[(1,1,3-trioxo-1,2-benzothiazole -5-yl) amino] pyrido [ 2,3-d] pyrimidine-2,4,7-trione

To a solution of 5-amino-1,1-dioxo-1,2-benzothiazol-3-one (134.81 mg, 646.16 μmol, 95.0%, 2.01 eq) in DMF (3 mL) was added NaH (51.44 mg, 1.29 mg). mmol, 60.0% purity, 4 eq) was added, stirred for 30 min at 0°C, and [3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8 at 0°C. -Dimethyl-2,4,7-trioxo-pyrido[2,3-d]pyrimidin-5-yl]trifluoromethanesulfonate (200 mg, 321.47 μmol, 98.9%, 1 eq) was added. . The reaction mixture was slowly warmed to 25°C and stirred at 25°C for 3 hours. 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 (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: Boston Prime C18 150*30 mm*5 um; Mobile phase: purified with [water(NH ₃ H ₂ O + NH ₄ HCO ₃ )-ACN]; B%: 28%-58%, 10 minutes) and then lyophilized to produce 3-cyclopropyl-1- (2-fluoro-4-iodo-phenyl)-6,8-dimethyl-5-[(1,1,3-trioxo-1,2-benzothiazol-5-yl)amino]pyrido[ 2,3-d]pyrimidine-2,4,7-trione (25 mg, 37.68 μmol, 11.7% yield, 100.0% purity) was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.93 (d, J = 8.1 Hz, 1H), 7.84 (s, 1H), 7.79 (d, J = 8.1 Hz, 1H), 7.66 (d, J = 10.2 Hz, 1H), 7.57 (d, J = 8.5 Hz, 1H), 6.89 (t, J = 8.5 Hz, 1H), 3.21 (s, 3H), 2.76-2.68 (m, 1H), 1.34 (s, 3H), 1.10-1.01 (m, 2H), 0.81-0.73 (m, 2H); ES-LCMS m/z 663.9 [M+H] ⁺ .

Step 6 : 3 - Cyclopropyl -5- (2- Fluoro -4- Iodo - Anilino ) -6,8-dimethyl-1- (1,1,3-trioxo-1,2-benzothiazole -5-yl) pyrido [4,3-d] pyrimidine-2,4,7-trione

3-Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-5-[(1,1,3-trioxo-1,2-) in THF (0.5 mL) A solution of benzothiazol-5-yl)amino]pyrido[2,3-d]pyrimidine-2,4,7-trione (20 mg, 30.15 μmol, 1 eq) in NaOMe (5.82 mg, 30.15 μmol) , 28.0% purity, 1 eq) was added. The mixture was stirred at 25°C for 1 hour. The reaction mixture was diluted with water (20 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 under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um; mobile phase: purified with [water(NH ₃ H ₂ O + NH ₄ HCO ₃ )-ACN]; B%: 29%-59%, 10 min) and then lyophilized to give 3-cyclopropyl-5- (2-fluoro-4-iodo-anilino)-6,8-dimethyl-1-(1,1,3-trioxo-1,2-benzothiazol-5-yl)pyrido [4, 3-d]pyrimidine-2,4,7-trione (2.99 mg, 4.46 μmol, 14.8% yield, 99.0% purity) was obtained as a white solid. ^1H NMR (500 MHz, CDCl ₃ ) δ ppm 11.06 (s, 1H), 7.80 (d, J = 7.9 Hz, 1H), 7.73 (s, 1H), 7.60 (d, J = 8.1 Hz, 1H), 7.47 (d, J = 10.4 Hz, 1H), 7.42 (d, J = 8.5 Hz, 1H), 6.70 (t, J = 8.3 Hz, 1H), 3.07 (s, 3H), 2.64 (s, 1H), 1.18 (s, 3H), 1.02 (d, J = 6.1 Hz, 2H), 0.70 (s, 2H); ES-LCMS m/z 663.9 [M+H] ⁺ .

I-102

Step 1 : tert -Butyl N- [3-[[3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyri nor[2,3-d]pyrimidin-5-yl]amino]-1-bicyclo[1.1.1]fentanyl]carbamate

N- [3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2, 3-d]pyrimidin-5-yl]2,6-lutidine (155.17 mg, 1.45 mmol, 168.66 μL, 3) in a solution of trifluoromethanesulfonate (300 mg, 482.70 μmol, 99.0%, 1 eq). eq) and tert -butyl N -(3-amino-1-bicyclo[1.1.1]fentanyl)carbamate (143.55 mg, 724.05 μmol, 1.5) were added. The mixture was stirred at 60°C for 2 hours. The reaction mixture was quenched by addition of H ₂ O (20 mL) and extracted using EtOAc (30 mL x 3). 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 analyzed by preparative TLC (PE/EtOAc = 2/1, TLC: PE /EtOAc = 2/1, R _f = 0.20) to give tert -butyl N -[3-[[3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8- Dimethyl-2,4,7-trioxo-pyrido[2,3- d ]pyrimidin-5-yl]amino]-1-bicyclo[1.1.1]fentanyl]carbamate (200 mg, 292.4 μmol, 60.6% yield, 97.0% purity) was obtained as a white solid. ^1H NMR (500 MHz, DMSO- d6 ) δ ppm 8.64 (s, 1H), 8.02-7.87 (m, 1H), 7.76-7.63 (m, 1H), _7.54 (s, 1H), 7.25-7.13 ( m, 1H), 2.74-2.67 (m, 3H), 2.67-2.61 (m, 1H), 2.18-2.07 (m, 6H), 1.99-1.93 (m, 3H), 1.43-1.31 (m, 9H), 1.08-0.94 (m, 2H), 0.73-0.59 (m, 2H); ES-LCMS m/z 664.5 [M+H] ⁺ .

Step 2 : tert -Butyl N- [3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-2,4,7-trioxo-pyri [4,3- d ]pyrimidin-1-yl]-1-bicyclo[1.1.1]fentanyl]carbamate

N -[3-[[3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyri in THF (3 mL) [2,3- d ]pyrimidin-5-yl]amino]-1-bicyclo[1.1.1]fentanyl]carbamate (100 mg, 146.20 μmol, 97.0%, 1 eq) and NaOMe/MeOH (19.40 μmol) mg, 20 μL, 30.0%, 1.00 eq) was degassed and purged three times with N ₂ , and then the mixture was stirred at 25° C. for 1 hour under N ₂ atmosphere. The reaction mixture was quenched by addition of CH ₃ COOH (0.1 mL), then diluted with H ₂ O (20 mL) and extracted using EtOAc (30 mL x 3). 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 analyzed by preparative TLC (PE/EtOAc = 3/1, TLC: PE /EtOAc = 3/1, R _f = 0.30) to give tert -butyl N -[3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8- Dimethyl-2,4,7-trioxo-pyrido[4,3- d ]pyrimidin-1-yl]-1-bicyclo[1.1.1]fentanyl]carbamate (50 mg, 67.8 μmol, 46.4% Yield, 90.0% purity) was obtained as a white solid. ¹ H NMR (500 MHz, DMSO- d ₆ ) δ ppm 10.36-10.16 (m, 1H), 7.83-7.68 (m, 1H), 7.65-7.55 (m, 1H), 7.54-7.48 (m, 1H), 6.96-6.82 (m, 1H), 3.08-3.06 (m, 3H), 2.59-2.54 (m, 3H), 2.44-2.39 (m, 2H), 2.23-2.18 (m, 2H), 2.03-2.01 (m) , 3H), 1.39-1.35 (m, 9H), 0.93-0.84 (m, 2H), 0.63-0.39 (m, 2H); ES-LCMS m/z 664.0 [M+H] ⁺ .

Step 3 : tert -Butyl 1-(3-amino-1-bicyclo[1.1.1]fentanyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8 -dimethyl-pyrido[4,3- d ]pyrimidine-2,4,7-trione

tert -Butyl 1-(3-amino-1-bicyclo[1.1.1]fentanyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)- in DCM (1 mL) TFA (462.00 mg, 4.05 mmol, 0.3 mL, 59.74 eq) was added. The mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated to give tert -butyl 1-(3-amino-1-bicyclo[1.1.1]fentanyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6. ,8-dimethyl-pyrido[4,3- d ]pyrimidine-2,4,7-trione (30 mg, 44.29 μmol, 65.3% yield, 100.0% purity, TFA) was obtained as a white solid, which was It was used in the next step without further purification. ¹ H NMR (500 MHz, DMSO- d ₆ ) δ ppm 10.31 (s, 1H), 8.69 (s, 2H), 7.76 (dd, J=1.8, 10.2 Hz, 1H), 7.53 (d, J=8.5 Hz) , 1H), 6.90 (t, J=8.5 Hz, 1H), 3.06 (s, 3H), 2.65-2.62 (m, 1H), 2.52-2.51 (m, 6H), 2.01 (s, 3H), 0.94- 0.85 (m, 2H), 0.67-0.34 (m, 2H); ES-LCMS m/z 564.1 [M+H] ⁺ .

Step 3 : tert -Butyl 3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-1-[3-(methylsulfamoylamino)-1-bicyclo [1.1.1]fentanyl]pyrido[4,3- d ]pyrimidine-2,4,7-trione

1-(3-Amino-1-bicyclo[1.1.1]fentanyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8 in DCM (4 mL) -Dimethyl-pyrido[4,3- d ]pyrimidine-2,4,7-trione (30 mg, 44.29 μmol, 100.0%, 1 eq, TFA) in solution of TEA (10.78 mg, 106.50 μmol, 14.82 μmol) μL, 2 eq) and N -methylsulfamoyl chloride (6.90 mg, 53.25 μmol, 1 eq) were added. The mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated to obtain a residue, which was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um; mobile phase: [water (FA)-ACN]; B%: 51%-71%, 12 minutes) and then lyophilized to obtain the product, which was then lyophilized to produce tert -butyl 3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8- Dimethyl-1-[3-(methylsulfamoylamino)-1-bicyclo[1.1.1]fentanyl]pyrido[4,3-d]pyrimidine-2,4,7-trione (12.23 mg, 18.63 μmol, 42.1% yield, 100.0% purity) was obtained as a white solid. ^1H NMR (400 MHz, DMSO- d ₆ ) δ ppm 10.26 (s, 1H), 7.85 (s, 1H), 7.76 (dd, J = 1.8, 10.3 Hz, 1H), 7.52 (d, J = 8.6 Hz) , 1H), 6.88 (t, J = 8.6 Hz, 1H), 6.76 (q, J = 5.0 Hz, 1H), 3.06 (s, 3H), 2.57-2.54 (m, 1H), 2.52 (s, 1H) , 2.42 (d, J = 5.0 Hz, 6H), 2.23 (s, 3H), 2.02 (s, 3H), 0.88 (s, 2H), 0.67-0.32 (m, 2H); ES-LCMS m/z 657.0 [M+H]+.

I-103

Step 1 : tert -Butyl 3-[[3- cyclopropyl -1-(2- fluoro- 4- iodo -phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2 ,3-d]pyrimidin-5-yl]amino]azetidine-1-carboxylate

[3-Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido in 1,4-dioxane (8 mL) [2,3-d]pyrimidin-5-yl]trifluoromethanesulfonate (400 mg, 642.95 μmol, 98.9% purity, 1 eq) and tert -butyl 3-aminoazetidine-1-carboxylate (221.46 mg, 1.29 mmol, 2 eq) was added to a solution of Cs ₂ CO ₃ (628.45 mg, 1.93 mmol, 3 eq). The mixture was stirred at 40°C for 3 hours. TLC (petroleum ether:EtOAc=1:1, R _f = 0.4) showed that new spots had formed and the starting material was completely consumed. The reaction mixture was quenched by addition of H ₂ O (20 mL) and extracted using EtOAc (30 mL x 3). 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 analyzed by preparative TLC (PE/EtOAc = 1/1.5, TLC: PE/ EtOAc = 1/1, R _f = 0.40) to give tert -butyl 3-[[3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2, 4,7-trioxo-pyrido[2,3-d]pyrimidin-5-yl]amino]azetidine-1-carboxylate (150 mg, 223.55 μmol, 34.8% yield, 95.0% purity) was obtained as a white solid. was obtained. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.19 (d, J = 6.4 Hz, 1H), 7.61 (dd, J = 8.8, 15.6 Hz, 2H), 6.89 (t, J = 8.0 Hz, 1H), 4.56-4.46 (m, 1H), 4.27 (td, J = 8.4, 13.2 Hz, 2H), 3.92 (dt, J = 5.2, 9.2 Hz, 2H), 2.89 (s, 3H), 2.79-2.71 (m, 1H), 2.04 (s, 3H), 1.46 (s, 9H), 1.22-1.15 (m, 2H), 0.86-0.78 (m, 2H); ES-LCMS m/z 638.0 [M+H] ⁺ .

Step 2 : tert -Butyl 3-[[3-( cyclopropylcarbamoyl )-2-(2- fluoro- 4- iodo - anilino )-1,5-dimethyl-6-oxo-4-pyridyl ]amino]azetidine-1-carboxylate and tert -butyl 3-[3- cyclopropyl -5-(2- fluoro- 4- iodo - anilino )-6,8-dimethyl-2,4,7 - trioxo -pyrido[4,3-d]pyrimidin-1-yl]azetidine-1-carboxylate

tert -Butyl 3-[[3-cyclopropyl-1-(2-fluoro-) in a solution of NaOMe (13.42 mg, 74.52 μmol, 0.5 mL, 30% purity, 1 eq) in THF (30 mL) at 0°C. 4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2,3-d]pyrimidin-5-yl]amino]azetidine-1-carboxylate (50 mg, 74.52 μmol, 95% purity, 1 eq) was added. The mixture was stirred at 0°C for 1 hour. TLC (petroleum ether:EtOAc=3:1, R _f = 0.7) showed that a new spot was formed and the starting material was completely consumed. The reaction mixture was quenched by addition of AcOH (1 mL) and stirred at 0°C for 10 min. The mixture was quenched by addition of H ₂ O (20 mL) and extracted using EtOAc (30 mL x 3). 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. The residue was purified by preparative TLC (PE/EtOAc = 2/1, TLC: PE/EtOAc = 3/1, R _f = 0.70) to give tert -butyl 3-[[3-(cyclopropylcarbamoyl)-2. -(2-Fluoro-4-iodo-anilino)-1,5-dimethyl-6-oxo-4-pyridyl]amino]azetidine-1-carboxylate (20 mg, 31.07 μmol, 41.7% yield , 95.0% purity) and tert -butyl 3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-2,4,7-trioxo-pyr. Do[4,3-d]pyrimidin-1-yl]azetidine-1-carboxylate (20 mg, 29.81 μmol, 40.0% yield, 95.0% purity) was obtained as a white solid. ^1H NMR (500 MHz, CDCl ₃ ) δ ppm 10.87 (s, 1H), 7.50 (dd, J = 1.5, 9.5 Hz, 1H), 7.44 (d, J = 8.5 Hz, 1H), 6.68 (t, J = 8.5 Hz, 1H), 4.75 (quin, J =6.5 Hz, 1H), 4.26 - 4.15 (m, 4H), 3.19 - 3.12 (m, 3H), 2.71 (tt, J =4.0, 7.0 Hz, 1H) , 2.03 (s, 3H), 1.43 (s, 9H), 1.10 (q, J = 7.0 Hz, 2H), 0.73-0.67 (m, 2H); ^1H NMR (500 MHz, CDCl ₃ ) δ ppm 8.83 (s, 1H), 7.42 (dd, J = 2.0, 10.0 Hz, 1H), 7.30 (d, J = 8.5 Hz, 1H), 7.14 (d, J = 2.0 Hz, 1H), 6.35 (t, J = 8.5 Hz, 1H), 4.61 (d, J = 10.0 Hz, 1H), 4.21 (dd, J = 8.0, 9.0 Hz, 2H), 3.73 (dd, J = 5.0, 9.5 Hz, 2H), 3.25 (s, 3H), 2.74 (dt, J = 3.5, 7.0 Hz, 1H), 2.08 (s, 3H), 2.04 (s, 3H), 1.44 (s, 9H) , 0.80-0.75 (m, 2H), 0.38-0.32 (m, 2H); ES-LCMS m/z 612.1 [M+H] ⁺ ; ES-LCMS m/z 638.0 [M+H] ⁺ .

Step 3 : 1 -( azetidin -3-yl)-3- cyclopropyl -5-(2- fluoro- 4- iodo - anilino )-6,8-dimethyl-pyrido[4,3-d ]Pyrimidine-2,4,7-trione

tert -Butyl 3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-2,4,7-trioxo-pyri in DCM (5 mL) TFA (1.54 g, 13.51 mmol, 1 mL) in a solution of [4,3-d]pyrimidin-1-yl]azetidine-1-carboxylate (20 mg, 29.81 μmol, 95.0% purity, 1 eq). was added. The mixture was stirred at 25°C for 1 hour. The mixture was concentrated to 1-(azetidin-3-yl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-pyrido[4,3- d]pyrimidine-2,4,7-trione (16 mg, 28.29 μmol, 94.9% yield, 95.0% purity) was obtained as a white solid, which was used directly in the next step without further purification. ES-LCMS m/z 538.0 [M+H] ⁺ .

Step 4 : 3 -[3- Cyclopropyl -5- (2- Fluoro -4- Iodo - Anilino ) -6,8-dimethyl-2,4,7-trioxo-pyrido [4,3- d]pyrimidin-1-yl]- N -methyl-azetidine-1-sulfonamide

1-(azetidin-3-yl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8- in DCM (5 mL) at 0°C under N ₂ atmosphere. Dimethyl-pyrido[4,3-d]pyrimidine-2,4,7-trione (16 mg, 28.29 μmol, 95% purity, 1 eq) was dissolved in Et ₃ N (8.59 mg, 84.86 μmol, 11.81 μmol). μL, 3 eq) and N -methylsulfamoyl chloride (3.67 mg, 28.29 μmol, 1 eq) were added. The mixture was stirred at 0°C for 1 hour. The reaction mixture was quenched by addition of H ₂ O (20 mL) and extracted using EtOAc (30 mL x 3). 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 subjected to preparative HPLC (column: Welch Xtimate C18 150 * 25 mm * 5 μm). ; Mobile phase: [Water(NH ₄ HCO ₃ )-ACN]; B%: 36%-66%, 11 min) to give 3-[3-cyclopropyl-5-(2-fluoro-4-io) do-anilino)-6,8-dimethyl-2,4,7-trioxo-pyrido[4,3-d]pyrimidin-1-yl] -N -methyl-azetidine-1-sulfonamide ( 6.76 mg, 10.47 μmol, 37.0% yield, 100.0% purity) was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.64 (d, J = 10.0 Hz, 1H), 7.55 (d, J = 8.5 Hz, 1H), 6.84 (t, J = 8.5 Hz, 1H), 4.95 (d, J = 7.5 Hz, 1H), 4.08 (quin, J = 8.0 Hz, 4H), 3.18 (s, 3H), 2.72-2.67 (m, 4H), 2.05 (s, 3H), 1.03 (q, J = 7.0 Hz, 2H), 0.68-0.62 (m, 2H); ES-LCMS m/z 630.9 [M+H] ⁺ .

I-109

Step 1 : 3 - Cyclopropyl -1- (2- Fluoro -4- Iodophenyl ) -6,8-dimethyl-2,4,7- Trioxo -1,2,3,4,7,8- Hexahydropyrido[2,3- d ]pyrimidin-5-yl trifluoromethanesulfonate

3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-5-hydroxy-6,8-dimethyl-pyrido[2, 3- d ]trifluoride in a solution of pyrimidine-2,4,7-trione (5 g, 10.35 mmol, 1 eq) and 2,6-lutidine (1.88 g, 17.59 mmol, 2.05 mL, 1.7 eq). Romethylsulfonyl trifluoromethanesulfonate (5.84 g, 20.69 mmol, 3.41 mL, 2 eq) was added dropwise. The mixture was stirred at 25°C for 12 hours under N ₂ atmosphere. The reaction mixture was quenched with saturated aqueous NaHCO ₃ (40 mL x 3) and extracted using DCM (50 mL x 3). The organic layer was washed with 1N HCl (40 mL x 3) and brine (50 mL x 3), dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give a residue which was subjected to flash silica gel chromatography. (from EtOAc/DCM = 100/1 to 8/1, TLC: PE/EtOAc = 1/1, R _f = 0.3) to give 3-cyclopropyl-1-(2-fluoro-4-iodophenyl )-6,8-dimethyl-2,4,7-trioxo-1,2,3,4,7,8-hexahydropyrido[2,3- d ]pyrimidin-5-yltrifluoromethane The sulfonate (4 g, 6.43 mmol, 97.1% yield, 98.9% purity) was obtained as a white solid. ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 7.95 (dd, J = 1.2, 9.5 Hz, 1H), 7.72 (d, J = 8.6 Hz, 1H), 7.30 (t, J = 8.1 Hz, 1H ₎ ), 2.76 (s, 3H), 2.70-2.66 (m, 1H), 2.01 (s, 3H), 1.08-0.98 (m, 2H), 0.62 (t, J = 11.9 Hz, 2H); ES-LCMS m/z 615.9 [M+H] ⁺ .

Step 2 : tert -Butyl (3-((3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-2,4,7-trioxo-1,2,3,4 ,7,8-hexahydropyrido[2,3- d ]pyrimidin-5-yl)amino)cyclobutyl)carbamate

3-Cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-2,4,7-trioxo-1,2,3,4,7 in DMF (5 mL) , 8-hexahydropyrido [2,3- d ] pyrimidin-5-yl trifluoromethanesulfonate (350 mg, 563.15 μmol, 99.0%, 1 eq), tert -butyl N - (3-aminocyclo A mixture of butyl)carbamate (115.38 mg, 619.46 μmol, 1.1 eq) and Cs ₂ CO ₃ (550.45 mg, 1.69 mmol, 3 eq) was stirred at 40°C for 1 hour, and then the mixture was incubated for 25 minutes under N ₂ atmosphere. It was stirred at ℃ for 4 hours. The reaction mixture was diluted with H ₂ O (30 mL), extracted using EtOAc (40 mL x 3) and washed with brine (40 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was purified by prep-TLC (SiO ₂ , PE/EtOAc = 1/1, R _f = 0.3) to give tert -Butyl (3-((3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-2,4,7-trioxo-1,2,3,4, 7,8-Hexahydropyrido[2,3- d ]pyrimidin-5-yl)amino)cyclobutyl)carbamate (200 mg, 255.42 μmol, 45.4% yield, 83.2% purity) was obtained as a yellow solid. . ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 9.07 (d, J = 6.6 Hz, 1H), 7.88 (d, J = 9.3 Hz, 1H), 7.66 (d, J = 8.3 Hz _, 1H), 7.28 (d, J = 7.8 Hz, 1H), 7.16-7.11 (m, 1H), 4.27 (d, J = 6.4 Hz, 1H), 3.87-3.41 (m, 1H), 2.62 (s, 3H), 2.35 -2.09 (m, 4H), 1.95 (s, 3H), 1.93 (s, 1H), 1.34 (s, 9H), 0.99 (s, 2H), 0.65 (s, 2H); ES-LCMS m/z 652.1 [M+H] ⁺ .

Step 3 : tert -Butyl (3-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo-3,4,6 ,7-tetrahydropyrido[4,3- d ]pyrimidin-1(2H ) -yl)cyclobutyl)carbamate

N₂ in THF (30 mL) at 0°C under atmosphere.tert-Butyl (3-((3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-2,4,7-trioxo-1,2,3,4, 7,8-hexahydropyrido[2,3-d]NaOMe/MeOH (191.57 μmol, 0.25 mL, 30%, 1 eq) was added to a solution of pyrimidin-5-yl)amino)cyclobutyl)carbamate (150 mg, 191.57 μmol, 83.2%, 1 eq). . The mixture is N₂ The mixture was stirred for 0.5 hours at 25°C in an atmosphere. CH in residue until pH=7₃COOH was added and extracted using EtOAc (30 mL x 3). The combined organic layers were Na₂SO₄ dried, filtered, and concentrated under reduced pressure to give a residue, which was preparative-TLC (SiO₂, PE/EtOAc = 1:1, R_f1 = 0.3) and refined totert-Butyl(3-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo-3,4,6, 7-Tetrahydropyrido[4,3-d]Pyrimidine-1(2H)-yl)cyclobutyl)carbamate (30 mg, 32.56 μmol, 17.0% yield, 70.7% purity) was obtained as a yellow solid.^OneH NMR (500 MHz, CDCl₃)δ ppm 10.99-10.67 (m, 1H), 7.51 (d,J = 9.6 Hz, 1H), 7.45-7.41 (m, 1H), 6.68-6.61 (m, 1H), 4.09-3.93 (m, 1H), 3.76 (s, 3H), 3.53-3.41 (m, 1H), 3.17 (s, 3H), 3.12-2.99 (m, 1H), 2.79-2.70 (m, 2H), 2.48-2.24 (m, 2H), 1.45 (s, 9H), 1.18-1.07 (m, 2H), 0.92-0.86 (m, 2H); ES-LCMS m/z 652.1 [M+H]+.

Step 4 : 1 -(3- Aminocyclobutyl )-3- cyclopropyl -5-((2- fluoro -4- iodophenyl )amino)-6,8-dimethylpyrido[4,3- d ] Pyrimidine-2,4,7(1 H ,3 H ,6 H )-trione

tert -Butyl (3-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo in DCM (2 mL) -3,4,6,7-Tetrahydropyrido[4,3- d ]pyrimidin-1( 2H )-yl)cyclobutyl)carbamate (30 mg, 32.56 μmol, 70.7%, 1 eq) TFA (616.00 mg, 5.40 mmol, 0.4 mL, 165.94 eq) was added to the solution. The mixture was stirred at 25°C for 0.5 hours. The reaction mixture was diluted with H ₂ O (10 mL), extracted using DCM (30 mL x 3) and washed with brine (30 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give 1-(3-aminocyclobutyl)-3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino) -6,8-dimethylpyrido[4,3- d ]pyrimidine-2,4,7( 1H , 3H , 6H )-trione (30 mg, 31.56 μmol, 96.9% yield, 70.0% purity , TFA) was obtained as a yellow oil, which was used in the next step without further purification. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.62 (d, J = 10.5 Hz, 1H), 7.52 (d, J = 7.9 Hz, 1H), 6.87-6.74 (m, 1H), 5.33 (s, 2H), 4.16 (d, J = 7.0 Hz, 1H), 3.43 (d, J = 1.5 Hz, 1H), 3.16 (s, 3H), 2.72 (s, 2H), 2.64 (s, 1H), 2.28- 2.18 (m, 2H), 1.27 (s, 3H), 1.04-0.99 (m, 2H), 0.90-0.88 (m, 2H); ES-LCMS m/z 552.1 [M+H]+.

Step 5 : 3 - Cyclopropyl -5- (2- Fluoro -4- Iodo - Anilino ) -6,8-dimethyl-1- [3- (methylsulfamoylamino) cyclobutyl] pyrido [4, 3- d ]pyrimidine-2,4,7-trione

1-(3-Aminocyclobutyl)-3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethylpyrido[4,3] in DCM (2 mL) - d ] TEA (9.58 mg , 94.68 μmol, 13.18 μL, 3 eq) and N -methylsulfamoyl chloride (4.50 mg, 34.72 μmol, 1.1 eq) were added. The mixture was stirred at 25°C for 1 hour. The reaction mixture was diluted with H ₂ O (20 mL) and extracted using DCM (20 mL x 3). The organic layer was concentrated under reduced pressure and subjected to preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (NH ₄ HCO ₃ )-ACN]; B%: 45%-70%, 11 minutes) and lyophilized to obtain 3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-1-[3-(methylsulfamoylamino)cyclobutyl. ]pyrido[4,3- d ]pyrimidine-2,4,7-trione (3.13 mg, 4.81 μmol, 15.3% yield, 99.1% purity) was obtained as a yellow solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.64 (dd, J = 1.8, 10.0 Hz, 1H), 7.54 (d, J = 8.1 Hz, 1H), 6.82 (t, J = 8.5 Hz, 1H) , 4.29-4.20 (m, 1H), 3.51-3.47 (m, 1H), 3.18 (s, 3H), 2.87-2.82 (m, 2H), 2.67 (td, J = 3.4, 7.1 Hz, 1H), 2.58 (s, 3H), 2.44-2.37 (m, 2H), 2.08 (s, 3H), 1.05-0.98 (m, 2H), 0.66-0.62 (m, 2H); ES-LCMS m/z 645.0 [M+H] ⁺ .

I-111

Step 1 : 3 - Cyclopropyl -1- (2- Fluoro -4- Iodophenyl ) -6,8-dimethyl-5- ( (1- oxoisoindolin -5-yl) amino) pyrido [2, 3- d ]pyrimidine-2,4,7(1 H ,3 H ,8 H )-trione

[3-Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2,3-] in DMF (3 mL) [d ]pyrimidin-5-yl]5-aminoisoindolin-1-one (47.68 mg, 321.80 μmol, 1 eq) in a solution of trifluoromethanesulfonate (200 mg, 321.80 μmol, 99.0%, 1 eq) and Cs ₂ CO ₃ (314.54 mg, 965.39 μmol, 3 eq) were added. The mixture was stirred at 60°C for 3 hours. The reaction mixture was diluted with H ₂ O (30 mL), extracted using DCM (40 mL x 3), and the organic layer was concentrated under reduced pressure and preparative HPLC (column: Boston Prime C18 150*30 mm* 5 um; mobile phase: [water(NH ₃ H ₂ O + NH ₄ HCO ₃ )-ACN]; B%: 40%-70%, 10 min) and lyophilized to give 3-cyclopropyl-1-( 2-fluoro-4-iodophenyl)-6,8-dimethyl-5-((1-oxoisoindolin-5-yl)amino)pyrido[2,3- d ]pyrimidine-2,4, 7( 1H , 3H , 8H )-trione (120 mg, 193.09 μmol, 60.0% yield, 98.7% purity) was obtained as a white solid. ¹ H NMR (500 MHz, DMSO- d ₆ ) δ ppm 7.90-7.83 (m, 1H), 7.73-7.63 (m, 1H), 7.45 (d, J = 8.9 Hz, 1H), 6.79-6.62 (m, 2H), 6.06 (d, J = 6.6 Hz, 2H), 4.70-4.56 (m, 2H), 4.46 (d, J = 15.9 Hz, 1H), 3.38 (s, 3H), 2.52 (s, 1H), 2.50 (s, 3H), 2.07 (d, J = 1.5 Hz, 1H), 0.94-0.83 (m, 2H), 0.67-0.45 (m, 2H); ES-LCMS m/z 614.0 [M+H]+.

Step 2 : 3 - Cyclopropyl -5-((2- fluoro- 4- iodophenyl )amino)-6,8-dimethyl-1-(1-oxoisoindolin-5-yl)pyrido[4, 3- d ]pyrimidine-2,4,7(1 H ,3 H ,6 H )-trione

3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-5-((1-oxoisoindoline-5) in THF (30 mL) at 0°C under N ₂ atmosphere. -yl)amino)pyrido[2,3- d ]pyrimidine-2,4,7(1 H ,3 H ,8 H )-trione (30 mg, 48.27 μmol, 98.7%, 1 eq) solution MeONa/MeOH (48.27 μmol, 0.1 mL, 30%, 1 eq) was added. The resulting mixture was stirred at 25°C for 20 minutes under N ₂ atmosphere. CH ₃ COOH was added to the residue until pH = 7 and extracted using EtOAc (30 mL x 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered, concentrated under reduced pressure and preparative HPLC (column: Boston Prime C18 150*30 mm* 5 um; mobile phase: [water (TFA)-ACN]; B%: 38%-58%, 12 minutes) and lyophilized to give 3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-1-( 1-oxoisoindolin-5-yl)pyrido[4,3- d ]pyrimidine-2,4,7( 1H , 3H , 6H )-trione (3.84 mg, 6.26 μmol, 13.0% yield) , 100.0% purity) was obtained as a white solid. ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 11.01 (s, 1H), 8.46 (s, 1H), 7.87 (dd _, J = 1.5, 9.4 Hz, 1H), 7.70-7.63 (m, 2H) , 7.27-7.22 (m, 1H), 7.21-7.17 (m, 2H), 4.34 (s, 2H), 3.11 (s, 3H), 2.33 (d, J = 1.8 Hz, 1H), 1.29 (s, 3H) ), 0.95 (d, J = 7.0 Hz, 2H), 0.67-0.57 (m, 2H); ES-LCMS m/z 613.9 [M+H]+.

3- cyclopropyl -5-(2- fluoro -4- iodo - Anilino )-6,8-dimethyl-1-[3-[ Methyl(methylsulfamoyl)amino Synthesis of ]phenyl]-2,4,7-trioxo-pyrido[4,3-d]pyrimidine (I-55)

step 1: 3 - cyclopropyl -5-(2- fluoro -4- iodo - Anilino )-6,8-dimethyl-1- [3- (methylamino)phenyl]pyrido[4,3-d]pyrimidine-2,4,7-trione

1-(3-aminophenyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-pyrido[4,3-] in THF (5 mL) d ]To a solution of pyrimidine-2,4,7-trione (100 mg, 174.41 μmol, 100% purity, 1 eq) was added NaH (40 mg, 1.00 mmol, 60% purity, 5.73 eq). The mixture was stirred at 25°C for 0.5 hours. MeI (30 mg, 211.36 μmol, 13.16 μL, 1.21 eq) was added. The mixture was stirred at 25°C for 12 hours. The reaction mixture was quenched with H ₂ O (20 mL) and extracted using EtOAc (20 mL x 3). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was subjected to preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (HCl)] -ACN]; B%: 49%-69%, 10 minutes) and lyophilized to give 3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl- 1-[3-(methylamino)phenyl]pyrido[4,3-d]pyrimidine-2,4,7-trione (6 mg, 9.87 μmol, 5.7% yield, 96.6% purity) was purified as an off-white solid. Obtained. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.68 (dd, J = 1.8, 10.2 Hz, 1H), 7.66-7.62 (m, 1H), 7.60 (d, J = 8.4 Hz, 1H), 7.50 (t, J = 1.9 Hz, 1H), 7.47-7.40 (m, 2H), 6.90 (t, J = 8.5 Hz, 1H), 3.22 (s, 3H), 3.10 (s, 3H), 2.74 (tt, J = 3.7, 7.2 Hz, 1H), 1.37 (s, 3H), 1.11-1.06 (m, 2H), 0.81-0.76 (m, 2H); ES-LCMS m/z 588.1 [M+H] ⁺

step 2: 3 - cyclopropyl -5-(2- fluoro -4- iodo - Anilino )-6,8-dimethyl-1-[3-[methyl(methylsulfamoyl)amino]phenyl]-2,4,7-trioxo-pyrido[4,3-d]pyrimidine

3-Cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-1-[3-(methylamino)phenyl]pyrido in DCM (5 mL) at 0°C. [4,3-d]pyrimidine-2,4,7-trione (5 mg, 8.22 μmol, 96.6% purity, 1 eq) and DIEA (22.26 mg, 172.23 μmol, 30 μL, 20.95 eq). N -methylsulfamoyl chloride (10 mg, 77.18 μmol, 9.39 eq) was added. The mixture was stirred at 0°C for 10 minutes. The reaction mixture was diluted with H ₂ O (20 mL) and extracted using DCM (30 mL x 3). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was subjected to preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (HCl)] -ACN]; B%: 54%-74%, 10 minutes) and lyophilized to produce 3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl- 1-[3-[methyl(methylsulfamoyl)amino]phenyl]-2,4,7-trioxo-pyrido[4,3-d]pyrimidine (1.58 mg, 2.28 μmol, 27.8% yield, 98.3% Purity) was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 11.24 (s, 1H), 7.68 (dd, J = 1.3, 10.3 Hz, 1H), 7.58 (d, J = 8.4 Hz, 1H), 7.53-7.48 (m, 2H), 7.48-7.44 (m, 1H) , 7.34-7.25 (m, 1H), 6.88 (dt, J = 2.5, 8.5 Hz, 1H), 3.27 (s, 3H), 3.22 (d, J = 2.9 Hz, 3H), 2.73 (tt, J = 3.7) , 7.2 Hz, 1H), 2.65 (s, 3H), 1.39 (s, 3H), 1.13-1.02 (m, 2H), 0.83-0.75 (m, 2H); ES-LCMS m/z 680.9 [M+H] ⁺ .

tert -Butyl 3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-1-[3-(methylsulfamoylamino)-1-bicyclo[1.1. Synthesis of 1]fentanyl]pyrido[4,3- d ]pyrimidine-2,4,7-trione (I-102)

Step 1: tert -Butyl N -[3-[[3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2,3-d ]pyrimidin-5-yl]amino]-1-bicyclo[1.1.1]fentanyl]carbamate

N- [3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2, 2,6-lutidine (155.17 mg, 1.45 mmol, 168.66 μL, 3) in a solution of 3-d]pyrimidin-5-yl]trifluoromethanesulfonate (300 mg, 482.70 μmol, 99.0%, 1 eq) eq) and tert -butyl N -(3-amino-1-bicyclo[1.1.1]fentanyl)carbamate (143.55 mg, 724.05 μmol, 1.5) were added. The mixture was stirred at 60°C for 2 hours. The reaction mixture was quenched by addition of H ₂ O (20 mL) and extracted using EtOAc (30 mL x 3). 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 analyzed by preparative TLC (PE/EtOAc = 2/1, TLC: PE /EtOAc = 2/1, R _f = 0.20) to give tert -butyl N -[3-[[3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8- Dimethyl-2,4,7-trioxo-pyrido[2,3- d ]pyrimidin-5-yl]amino]-1-bicyclo[1.1.1]fentanyl]carbamate (200 mg, 292.4 μmol, 60.6% yield, 97.0% purity) was obtained as a white solid. ^1H NMR (500 MHz, DMSO- d6 ) δ ppm 8.64 (s, 1H), 8.02-7.87 (m, 1H), 7.76-7.63 (m, 1H), _7.54 (s, 1H), 7.25-7.13 ( m, 1H), 2.74-2.67 (m, 3H), 2.67-2.61 (m, 1H), 2.18-2.07 (m, 6H), 1.99-1.93 (m, 3H), 1.43-1.31 (m, 9H), 1.08-0.94 (m, 2H), 0.73-0.59 (m, 2H); ES-LCMS m/z 664.5 [M+H] ⁺ .

Step 2: tert -butyl N -[3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-2,4,7-trioxo-pyrido[4,3- d ]pyrimidin-1-yl]-1-bicyclo[1.1.1]fentanyl]carbamate

N -[3-[[3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyri in THF (3 mL) [2,3- d ]pyrimidin-5-yl]amino]-1-bicyclo[1.1.1]fentanyl]carbamate (100 mg, 146.20 μmol, 97.0%, 1 eq) and NaOMe/MeOH (19.40 μmol) mg, 20 μL, 30.0%, 1.00 eq) was degassed and purged three times with N ₂ , and then the mixture was stirred at 25° C. for 1 hour under N ₂ atmosphere. The reaction mixture was quenched by addition of CH ₃ COOH (0.1 mL), then diluted with H ₂ O (20 mL) and extracted using EtOAc (30 mL x 3). 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 analyzed by preparative TLC (PE/EtOAc = 3/1, TLC: PE /EtOAc = 3/1, R _f = 0.30) to give tert -butyl N -[3-[3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8- Dimethyl-2,4,7-trioxo-pyrido[4,3- d ]pyrimidin-1-yl]-1-bicyclo[1.1.1]fentanyl]carbamate (50 mg, 67.8 μmol, 46.4% Yield, 90.0% purity) was obtained as a white solid. ¹ H NMR (500 MHz, DMSO- d ₆ ) δ ppm 10.36-10.16 (m, 1H), 7.83-7.68 (m, 1H), 7.65-7.55 (m, 1H), 7.54-7.48 (m, 1H), 6.96-6.82 (m, 1H), 3.08-3.06 (m, 3H), 2.59-2.54 (m, 3H), 2.44-2.39 (m, 2H), 2.23-2.18 (m, 2H), 2.03-2.01 (m) , 3H), 1.39-1.35 (m, 9H), 0.93-0.84 (m, 2H), 0.63-0.39 (m, 2H); ES-LCMS m/z 664.0 [M+H] ⁺ .

Step 3: tert -Butyl 1-(3-amino-1-bicyclo[1.1.1]fentanyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-pyri degree[4,3- d ]Pyrimidine-2,4,7-trione

tert -Butyl 1-(3-amino-1-bicyclo[1.1.1]fentanyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)- in DCM (1 mL) TFA (462.00 mg, 4.05 mmol, 0.3 mL, 59.74 eq) was added. The mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated to give tert -butyl 1-(3-amino-1-bicyclo[1.1.1]fentanyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6. , 8-dimethyl-pyrido [4,3- d ] pyrimidine-2,4,7-trione (30 mg, 44.29 μmol, 65.3% yield, 100.0% purity, TFA) was obtained as a white solid, which was added It was used in the next step without purification. ¹ H NMR (500 MHz, DMSO- d ₆ ) δ ppm 10.31 (s, 1H), 8.69 (s, 2H), 7.76 (dd, J=1.8, 10.2 Hz, 1H), 7.53 (d, J=8.5 Hz) , 1H), 6.90 (t, J=8.5 Hz, 1H), 3.06 (s, 3H), 2.65-2.62 (m, 1H), 2.52-2.51 (m, 6H), 2.01 (s, 3H), 0.94- 0.85 (m, 2H), 0.67-0.34 (m, 2H); ES-LCMS m/z 564.1 [M+H] ⁺ .

Step 3: tert -Butyl 3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-1-[3-(methylsulfamoylamino)-1-bicyclo[1.1.1 ]fentanyl]pyrido[4,3- d ]Pyrimidine-2,4,7-trione

1-(3-Amino-1-bicyclo[1.1.1]fentanyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8 in DCM (4 mL) -Dimethyl-pyrido[4,3- d ]pyrimidine-2,4,7-trione (30 mg, 44.29 μmol, 100.0%, 1 eq, TFA) in solution of TEA (10.78 mg, 106.50 μmol, 14.82 μmol) μL, 2 eq) and N -methylsulfamoyl chloride (6.90 mg, 53.25 μmol, 1 eq) were added. The mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated to obtain a residue, which was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um; mobile phase: [water (FA)-ACN]; B%: 51%-71%, 12 minutes) and then lyophilized to obtain the product, which was then lyophilized to produce tert -butyl 3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8- Dimethyl-1-[3-(methylsulfamoylamino)-1-bicyclo[1.1.1]fentanyl]pyrido[4,3-d]pyrimidine-2,4,7-trione (12.23 mg, 18.63 μmol, 42.1% yield, 100.0% purity) was obtained as a white solid. ^1H NMR (400 MHz, DMSO- d ₆ ) δ ppm 10.26 (s, 1H), 7.85 (s, 1H), 7.76 (dd, J = 1.8, 10.3 Hz, 1H), 7.52 (d, J = 8.6 Hz) , 1H), 6.88 (t, J = 8.6 Hz, 1H), 6.76 (q, J = 5.0 Hz, 1H), 3.06 (s, 3H), 2.57-2.54 (m, 1H), 2.52 (s, 1H) , 2.42 (d, J = 5.0 Hz, 6H), 2.23 (s, 3H), 2.02 (s, 3H), 0.88 (s, 2H), 0.67-0.32 (m, 2H); ES-LCMS m/z 657.0 [M+H]+.

3- Cyclopropyl -5- (2- Fluoro -4- Iodo - Anilino )-6,8-dimethyl-1- [3- ( Methylsulfamoylamino ) Cyclobutyl] pyrido [4,3- d ] Synthesis of pyrimidine-2,4,7-trione (I-109)

step 1: 3 - cyclopropyl -1-(2- fluoro -4- Iodophenyl )-6,8-dimethyl-2,4,7- Trioxo -1,2,3,4,7,8-hexahydropyrido[2,3- d ]Pyrimidine-5-yl Trifluoromethane sulfonate

3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-5-hydroxy-6,8-dimethyl-pyrido[2, 3- d ]trifluoride in a solution of pyrimidine-2,4,7-trione (5 g, 10.35 mmol, 1 eq) and 2,6-lutidine (1.88 g, 17.59 mmol, 2.05 mL, 1.7 eq). Romethylsulfonyl trifluoromethanesulfonate (5.84 g, 20.69 mmol, 3.41 mL, 2 eq) was added dropwise. The mixture was stirred at 25°C for 12 hours under N ₂ atmosphere. The reaction mixture was quenched with saturated aqueous NaHCO ₃ (40 mL x 3) and extracted using DCM (50 mL x 3). The organic layer was washed with 1N HCl (40 mL x 3) and brine (50 mL x 3), dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give a residue which was subjected to flash silica gel chromatography. (from EtOAc/DCM = 100/1 to 8/1, TLC: PE/EtOAc = 1/1, R _f = 0.3) to give 3-cyclopropyl-1-(2-fluoro-4-iodophenyl )-6,8-dimethyl-2,4,7-trioxo-1,2,3,4,7,8-hexahydropyrido[2,3- d ]pyrimidin-5-yltrifluoromethane The sulfonate (4 g, 6.43 mmol, 97.1% yield, 98.9% purity) was obtained as a white solid. ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 7.95 (dd, J = 1.2, 9.5 Hz, 1H), 7.72 (d, J = 8.6 Hz, 1H), 7.30 (t, J = 8.1 Hz, 1H ₎ ), 2.76 (s, 3H), 2.70-2.66 (m, 1H), 2.01 (s, 3H), 1.08-0.98 (m, 2H), 0.62 (t, J = 11.9 Hz, 2H); ES-LCMS m/z 615.9 [M+H] ⁺ .

Step 2: tert -Butyl (3-((3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-2,4,7-trioxo-1,2,3,4, 7,8-hexahydropyrido[2,3- d ]pyrimidin-5-yl)amino)cyclobutyl)carbamate

3-Cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-2,4,7-trioxo-1,2,3,4,7 in DMF (5 mL) , 8-hexahydropyrido [2,3- d ] pyrimidin-5-yl trifluoromethanesulfonate (350 mg, 563.15 μmol, 99.0%, 1 eq), tert -butyl N - (3-aminocyclo A mixture of butyl)carbamate (115.38 mg, 619.46 μmol, 1.1 eq) and Cs ₂ CO ₃ (550.45 mg, 1.69 mmol, 3 eq) was stirred at 40°C for 1 hour, and then the mixture was incubated for 25 minutes under N ₂ atmosphere. It was stirred at ℃ for 4 hours. The reaction mixture was diluted with H ₂ O (30 mL), extracted using EtOAc (40 mL x 3) and washed with brine (40 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was purified by prep-TLC (SiO ₂ , PE/EtOAc = 1/1, R _f = 0.3) to give tert -Butyl (3-((3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-2,4,7-trioxo-1,2,3,4, 7,8-Hexahydropyrido[2,3- d ]pyrimidin-5-yl)amino)cyclobutyl)carbamate (200 mg, 255.42 μmol, 45.4% yield, 83.2% purity) was obtained as a yellow solid. . ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 9.07 (d, J = 6.6 Hz, 1H), 7.88 (d, J = 9.3 Hz, 1H), 7.66 (d, J = 8.3 Hz _, 1H), 7.28 (d, J = 7.8 Hz, 1H), 7.16-7.11 (m, 1H), 4.27 (d, J = 6.4 Hz, 1H), 3.87-3.41 (m, 1H), 2.62 (s, 3H), 2.35 -2.09 (m, 4H), 1.95 (s, 3H), 1.93 (s, 1H), 1.34 (s, 9H), 0.99 (s, 2H), 0.65 (s, 2H); ES-LCMS m/z 652.1 [M+H] ⁺ .

Step 3: tert -Butyl (3-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo-3,4,6, 7-Tetrahydropyrido[4,3- d ]Pyrimidine-1(2 H )-yl)cyclobutyl)carbamate

tert -butyl (3-((3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-2,4 in THF (30 mL) at 0°C under N ₂ atmosphere. ,7-trioxo-1,2,3,4,7,8-hexahydropyrido [2,3- d ] pyrimidin-5-yl) amino) cyclobutyl) carbamate (150 mg, 191.57 μmol, NaOMe/MeOH (191.57 μmol, 0.25 mL, 30%, 1 eq) was added to the solution (83.2%, 1 eq). The mixture was stirred at 25°C for 0.5 hours under N ₂ atmosphere. CH ₃ COOH was added to the residue until pH = 7 and extracted using EtOAc (30 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 prep-TLC (SiO ₂ , PE/EtOAc = 1:1, R _f1 = 0.3). tert -Butyl(3-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo-3,4,6 , 7-Tetrahydropyrido[4,3- d ]pyrimidin-1( 2H )-yl)cyclobutyl)carbamate (30 mg, 32.56 μmol, 17.0% yield, 70.7% purity) was obtained as a yellow solid. did. ^1H NMR (500 MHz, CDCl ₃ ) δ ppm 10.99-10.67 (m, 1H), 7.51 (d, J = 9.6 Hz, 1H), 7.45-7.41 (m, 1H), 6.68-6.61 (m, 1H) , 4.09-3.93 (m, 1H), 3.76 (s, 3H), 3.53-3.41 (m, 1H), 3.17 (s, 3H), 3.12-2.99 (m, 1H), 2.79-2.70 (m, 2H) , 2.48-2.24 (m, 2H), 1.45 (s, 9H), 1.18-1.07 (m, 2H), 0.92-0.86 (m, 2H); ES-LCMS m/z 652.1 [M+H]+.

step 4: 1 -(3- Aminocyclobutyl )-3- cyclopropyl -5-((2- fluoro -4- Iodophenyl )Amino)-6,8-dimethylpyrido[4,3- d ]Pyrimidine-2,4,7(1 H ,3 H ,6 H ) - trion

tert -Butyl (3-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo in DCM (2 mL) -3,4,6,7-Tetrahydropyrido[4,3- d ]pyrimidin-1( 2H )-yl)cyclobutyl)carbamate (30 mg, 32.56 μmol, 70.7%, 1 eq) TFA (616.00 mg, 5.40 mmol, 0.4 mL, 165.94 eq) was added to the solution. The mixture was stirred at 25°C for 0.5 hours. The reaction mixture was diluted with H ₂ O (10 mL), extracted using DCM (30 mL x 3) and washed with brine (30 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give 1-(3-aminocyclobutyl)-3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino) -6,8-dimethylpyrido[4,3- d ]pyrimidine-2,4,7( 1H , 3H , 6H )-trione (30 mg, 31.56 μmol, 96.9% yield, 70.0% purity , TFA) was obtained as a yellow oil, which was used in the next step without further purification. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.62 (d, J = 10.5 Hz, 1H), 7.52 (d, J = 7.9 Hz, 1H), 6.87-6.74 (m, 1H), 5.33 (s, 2H), 4.16 (d, J = 7.0 Hz, 1H), 3.43 (d, J = 1.5 Hz, 1H), 3.16 (s, 3H), 2.72 (s, 2H), 2.64 (s, 1H), 2.28- 2.18 (m, 2H), 1.27 (s, 3H), 1.04-0.99 (m, 2H), 0.90-0.88 (m, 2H); ES-LCMS m/z 552.1 [M+H]+.

step 5: 3 - cyclopropyl -5-(2- fluoro -4- iodo - Anilino )-6,8-dimethyl-1- [3- (methylsulfamoylamino)cyclobutyl]pyrido[4,3- d ]Pyrimidine-2,4,7-trione

1-(3-Aminocyclobutyl)-3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethylpyrido[4,3] in DCM (2 mL) - d ] TEA (9.58 mg , 94.68 μmol, 13.18 μL, 3 eq) and N -methylsulfamoyl chloride (4.50 mg, 34.72 μmol, 1.1 eq) were added. The mixture was stirred at 25°C for 1 hour. The reaction mixture was diluted with H ₂ O (20 mL) and extracted using DCM (20 mL x 3). The organic layer was concentrated under reduced pressure and subjected to preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (NH ₄ HCO ₃ )-ACN]; B%: 45%-70%, 11 minutes) and lyophilized to obtain 3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-1-[3-(methylsulfamoylamino)cyclobutyl. ]pyrido[4,3- d ]pyrimidine-2,4,7-trione (3.13 mg, 4.81 μmol, 15.3% yield, 99.1% purity) was obtained as a yellow solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.64 (dd, J = 1.8, 10.0 Hz, 1H), 7.54 (d, J = 8.1 Hz, 1H), 6.82 (t, J = 8.5 Hz, 1H) , 4.29-4.20 (m, 1H), 3.51-3.47 (m, 1H), 3.18 (s, 3H), 2.87-2.82 (m, 2H), 2.67 (td, J = 3.4, 7.1 Hz, 1H), 2.58 (s, 3H), 2.44-2.37 (m, 2H), 2.08 (s, 3H), 1.05-0.98 (m, 2H), 0.66-0.62 (m, 2H); ES-LCMS m/z 645.0 [M+H] ⁺ .

3- Cyclopropyl -5-((2- Fluoro -4- Iodophenyl ) Amino)-6,8-dimethyl-1- (1-oxoisoindolin-5-yl) Pyrido [4,3-d ]Synthesis of pyrimidine-2,4,7(1H,3H,6H)-trione (I-116)

step 1: 3 - cyclopropyl -1-(2- fluoro -4- Iodophenyl )-6,8-dimethyl-5- ( (One- Oxoisoindoline -5-yl)amino)pyrido[2,3- d ]Pyrimidine-2,4,7(1 H ,3 H ,8 H ) - trion

[3-Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2,3-] in DMF (3 mL) [d ]pyrimidin-5-yl]5-aminoisoindolin-1-one (47.68 mg, 321.80 μmol, 1 eq) in a solution of trifluoromethanesulfonate (200 mg, 321.80 μmol, 99.0%, 1 eq) and Cs ₂ CO ₃ (314.54 mg, 965.39 μmol, 3 eq) were added. The mixture was stirred at 60°C for 3 hours. The reaction mixture was diluted with H ₂ O (30 mL), extracted using DCM (40 mL x 3), and the organic layer was concentrated under reduced pressure and preparative HPLC (column: Boston Prime C18 150*30 mm* 5 um; mobile phase: [water(NH ₃ H ₂ O + NH ₄ HCO ₃ )-ACN]; B%: 40%-70%, 10 min) and lyophilized to give 3-cyclopropyl-1-( 2-fluoro-4-iodophenyl)-6,8-dimethyl-5-((1-oxoisoindolin-5-yl)amino)pyrido[2,3- d ]pyrimidine-2,4, 7( 1H , 3H , 8H )-trione (120 mg, 193.09 μmol, 60.0% yield, 98.7% purity) was obtained as a white solid. ¹ H NMR (500 MHz, DMSO- d ₆ ) δ ppm 7.90-7.83 (m, 1H), 7.73-7.63 (m, 1H), 7.45 (d, J = 8.9 Hz, 1H), 6.79-6.62 (m, 2H), 6.06 (d, J = 6.6 Hz, 2H), 4.70-4.56 (m, 2H), 4.46 (d, J = 15.9 Hz, 1H), 3.38 (s, 3H), 2.52 (s, 1H), 2.50 (s, 3H), 2.07 (d, J = 1.5 Hz, 1H), 0.94-0.83 (m, 2H), 0.67-0.45 (m, 2H); ES-LCMS m/z 614.0 [M+H]+.

step 2: 3 - cyclopropyl -5-((2- fluoro -4- Iodophenyl )Amino)-6,8-dimethyl-1-(1-oxoisoindolin-5-yl)pyrido[4,3- d ]Pyrimidine-2,4,7(1 H ,3 H ,6 H ) - trion

3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-5-((1-oxoisoindoline-5) in THF (30 mL) at 0°C under N ₂ atmosphere. -yl)amino)pyrido[2,3- d ]pyrimidine-2,4,7(1 H ,3 H ,8 H )-trione (30 mg, 48.27 μmol, 98.7%, 1 eq) solution MeONa/MeOH (48.27 μmol, 0.1 mL, 30%, 1 eq) was added. The resulting mixture was stirred at 25°C for 20 minutes under N ₂ atmosphere. CH ₃ COOH was added to the residue until pH = 7 and extracted using EtOAc (30 mL x 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered, concentrated under reduced pressure, and preparative HPLC (column: Boston Prime C18 150*30 mm* 5 um; mobile phase: [water (TFA)-ACN]; B%: 38%-58%, 12 minutes) and lyophilized to give 3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-1-( 1-oxoisoindolin-5-yl)pyrido[4,3- d ]pyrimidine-2,4,7( 1H , 3H , 6H )-trione (3.84 mg, 6.26 μmol, 13.0% yield) , 100.0% purity) was obtained as a white solid. ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 11.01 (s, 1H), 8.46 (s, 1H), 7.87 (dd _, J = 1.5, 9.4 Hz, 1H), 7.70-7.63 (m, 2H) , 7.27-7.22 (m, 1H), 7.21-7.17 (m, 2H), 4.34 (s, 2H), 3.11 (s, 3H), 2.33 (d, J = 1.8 Hz, 1H), 1.29 (s, 3H) ), 0.95 (d, J = 7.0 Hz, 2H), 0.67-0.57 (m, 2H); ES-LCMS m/z 613.9 [M+H]+.

1-(3-(3- cyclopropyl -5-((2- fluoro- 4- iodophenyl )amino)-6,8-dimethyl-2,4,7-trioxo-3,4,6, Synthesis of 7-tetrahydropyrido[4,3- d ]pyrimidin-1(2H ) -yl)phenyl) -N -methylmethanesulfonamide (I-46)

Step 1: 4 - Methylbenzenesulfonate N - Methyl -1- (3- Nitrophenyl ) Methanesulfonamide

To a solution of (3-nitrophenyl)methanesulfonyl chloride (500 mg, 2.12 mmol, 1 eq) in THF (1 mL) was added DIEA (548.47 mg, 4.24 mmol, 739.17 μL, 2 eq) and MeNH ₂ .EtOH (197.69 mg, 6.37 mmol, 212.18 μL, 3 eq) was added. The mixture was stirred at 25°C for 1 hour. The mixture was quenched with water (30 mL) and extracted using DCM (30 mL x 3). The combined organic phases were washed with NaHCO ₃ (30 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 (PE/EtOAc = 100/1). From 1/1, purified by TLC: PE/EtOAc = 2/1, R _f = 0.4) to give 4-methylbenzenesulfonate N -methyl-1-(3-nitrophenyl)methanesulfonamide (200 mg, 861.19 μmol, 40.6% yield, 99.1% purity) was obtained as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.32-8.20 (m, 2H), 7.79 (d, J = 7.6 Hz, 1H), 7.64-7.58 (m, 1H), 4.35 (s, 2H), 4.27 -4.13 (m, 1H), 2.79 (d, J = 5.4 Hz, 3H).

Step 2: 1 -(3- Aminophenyl )- N - Methylmethanesulfonamide

Pd /C ₍ 200 mg, 861.19 μmol, 10.0%, 1 eq) was added. The suspension was degassed under vacuum and purged several times with H ₂ . The mixture was stirred at 25° C. under H ₂ (15 psi) for 1 hour. The reaction mixture was filtered and concentrated under reduced pressure to obtain 1-(3-aminophenyl)- N -methyl-methanesulfonamide (180 mg, 853.90 μmol, 99.2% yield, 95.0% purity) as a yellow solid, which was further purified. It was used in the next step without it. ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 7.22-7.17 (m, 1H), 7.17-7.14 (m, 1H), 6.75-6.73 (m, 2H), 6.71-6.67 (m, 2H), 6.66- 6.57 (m, 1H), 4.18 (s, 2H), 2.02 (s, 3H).

Step 3: 1-(3-((3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-2,4,7-trioxo-1,2,3 ,4,7,8-hexahydropyrido[2,3-d]pyrimidin-5-yl)amino)phenyl)- N -methylmethanesulfonamide

2,6-lutidine (119.77 mg, 1.12 mmol) in a solution of 1-(3-aminophenyl)- N -methyl-methanesulfonamide (150 mg, 711.58 μmol, 95.0%, 1.91 eq) in DMA (3 mL) , 130.19 μL, 3 eq) and [3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2, 3- d ]pyrimidin-5-yl] 4-methylbenzenesulfonate (250 mg, 372.60 μmol, 90.0%, 1 eq) was added. The mixture was stirred at 140° C. under a microwave (2 bar) for 3 hours. The mixture was diluted with water (30 mL) and extracted using ethyl acetate (30 mL x 3). The organic layer was dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (HCl)- ACN]; B%: 46%-66%, 10 min) and lyophilized to obtain the product, which was added with saturated aqueous NaHCO ₃ to pH = 7 and purified using EtOAc (20 mL x 3). After extraction, the combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give 1-(3-((3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6 ,8-dimethyl-2,4,7-trioxo-1,2,3,4,7,8-hexahydropyrido [2,3- d ] pyrimidin-5-yl) amino) phenyl) - N -Methylmethanesulfonamide (45 mg, 58.55 μmol, 15.7% yield, 86.6% purity) was obtained as a yellow solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.85 (dd, J = 1.8, 9.5 Hz, 1H), 7.76 (d, J = 8.2 Hz, 1H), 7.37 (t, J = 7.8 Hz, 1H) , 7.22-7.14 (m, 2H), 7.08-6.99 (m, 2H), 4.33 (s, 2H), 2.95 (s, 3H), 2.82-2.76 (m, 1H), 2.69 (s, 3H), 1.67 (s, 3H), 1.16-1.09 (m, 2H), 0.81 (d, J = 2.6 Hz, 2H); ES-LCMS m/z 666.0 [M+H] ⁺ .

Step 4: 1-(3-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo-3,4 ,6,7-Tetrahydropyrido[4,3- d ]pyrimidin-1(2H ) -yl)phenyl) -N -methylmethanesulfonamide

1-(3-((3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethyl-2,4,7-trioxo-1 in THF (1 mL) 2,3,4,7,8-hexahydropyrido [2,3- d ] pyrimidin-5-yl) amino) phenyl) - N -methylmethanesulfonamide (45 mg, 58.15 μmol, 86.0%, 1 To the solution of eq), one drop of NaOMe/MeOH (20 wt%, 58.15 μmol, 1 eq) was added. The mixture was stirred at 25°C for 1 hour under N ₂ atmosphere. AcOH was added to the residue until pH = 7 and extracted using EtOAc (20 mL x 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm* 5 um; mobile phase: [water (NH ₃ H ₂ O + NH ₄ HCO ₃ )-ACN]; B%: 53%-83%, 10 minutes) and lyophilized to obtain 1-(3-(3-cyclopropyl-5-((2-fluo Ro-4-iodophenyl) amino) -6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido [4,3- d ] pyrimidine-1 ( 2 H )-yl)phenyl)- N -methylmethanesulfonamide (10.01 mg, 14.96 μmol, 25.7% yield, 99.4% purity) was obtained as a white solid. ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 7.66 (dd, J = 1.7, 10.3 Hz, 1H), 7.57 (d, J = 8.3 Hz, 1H), 7.51-7.47 (m, 2H), 7.43 ( s, 1H), 7.39 (dd, J = 3.2, 5.6 Hz, 1H), 6.86 (t, J = 8.4 Hz, 1H), 4.37 (s, 2H), 3.20 (s, 3H), 2.76-2.69 (m , 1H), 2.67 (s, 3H), 1.34 (s, 3H), 1.08-1.03 (m, 2H), 0.81-0.74 (m, 2H); ES-LCMS m/z 666.1 [M+H] ⁺ .

6- Cyclopropyl -4- (2- Fluoro -4- Iodo - Anilino ) -3,7-dimethyl-8- [2-methyl-3- (methylsulfamoylamino) phenyl] pyrano [3, Synthesis of 2-c]pyridine-2,5- dione (I-33)

Step 1: 6 - Cyclopropyl -4- (2- Fluoro -4- Iodo - Anilino ) -3,7-dimethyl-8- [2-methyl-3- (methylsulfamoylamino) phenyl] pyrano [3,2-c]pyridine-2,5-dione

8-(3-Amino-2-methyl-phenyl)-6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-3,7-dimethyl in DCM (2 mL) at 0°C. -In a stirred solution of pyrano[3,2-c]pyridine-2,5-dione (50 mg, 70.01 μmol, 80% purity, 1 eq) and TEA (28.34 mg, 280.02 μmol, 38.98 μL, 4 eq) N - Methylsulfamoyl chloride (18.14 mg, 140.01 μmol, not measurable purity, 2 eq) was added. The reaction mixture was stirred at 0° C. for 5 hours under N ₂ atmosphere. The reaction mixture was quenched by adding saturated aqueous NaHCO ₃ (30 mL) at 0° C. and extracted using DCM (30 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 preparatively purified. Purified by HPLC (column: Boston Prime C18 150*30 mm* 5 um; mobile phase: [water(NH ₃ H ₂ O + NH ₄ HCO ₃ )-ACN]; B%: 53%-83%, 10 min) Then, freeze-dried to form 6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-3,7-dimethyl-8-[2-methyl-3-(methylsulfamoylamino)phenyl. ]Pyrano[3,2-c]pyridine-2,5-dione (6.22 mg, 9.36 μmol, 13.4% yield, 100.0% purity) was obtained as a white solid. ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 11.30 (s, 1H), 8.82 (s, 1H) _, 7.71 (d, J = 1.7, 10.3 Hz, 1H), 7.51 (d, J = 8.3 Hz) , 1H), 7.40 (d, J = 7.8 Hz, 1H), 7.27 (t, J = 7.7 Hz, 1H), 7.14 (q, J = 4.9 Hz, 1H), 7.01 (d, J = 7.6 Hz, 1H) ), 6.87 (t, J = 8.7 Hz, 1H), 3.03 (s, 1H), 2.55 (d, J = 5.1 Hz, 3H), 2.20 (s, 3H), 2.03 (s, 3H), 1.43 (s) , 3H), 1.23-1.17 (m, 2H), 0.96-0.84 (m, 2H); ES-LCMS m/z 665.0 [M+H] ⁺ .

tert -Butyl 3- Cyclopropyl -5- (2- Fluoro -4- Iodo - Anilino )-6,8-dimethyl-1-[4-(methylsulfamoylamino)phenyl]pyrido[4,3 - d ] Synthesis of pyrimidine-2,4,7-trione (I-82)

Step 1: tert -Butyl 5-(4- aminoanilino )-3- cyclopropyl -1-(2- fluoro- 4- iodo -phenyl)-6,8-dimethyl-pyrido[2,3- d ]pyrimidine-2,4,7-trione

[3-Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2,3-] in DMF (2 mL) d ]pyrimidin-5-yl]trifluoromethanesulfonate (300 mg, 482.70 μmol, 99.0%, 1 eq) in a solution of 2,6-lutidine (155.17 mg, 1.45 mmol, 168.66 μL, 3 eq) and Benzene-1,4-diamine (78.30 mg, 724.05 μmol, 1.5 eq) was added. The mixture was stirred at 60°C for 2 hours. The reaction mixture was quenched by addition of H ₂ O (20 mL) and extracted using EtOAc (30 mL x 3). 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 analyzed by preparative TLC (PE/EtOAc = 2/1, TLC: PE /EtOAc = 2/1, R _f = 0.20) to give tert -butyl 5-(4-aminoanilino)-3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6. ,8-Dimethyl-pyrido[2,3- d ]pyrimidine-2,4,7-trione (200 mg, 327.89 μmol, 67.9% yield, 94.0% purity) was obtained as a brown solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 10.16-10.03 (m, 1H), 8.04-7.87 (m, 1H), 7.77-7.60 (m, 1H), 7.32-7.13 (m, 1H), 6.85-6.65 (m, 2H), 6.63-6.45 (m, 2H), 5.24-4.99 (m, 2H), 2.73-2.67 (m, 3H), 1.51-1.34 (m, 3H), 1.09-0.89 (m) , 2H), 0.76-0.56 (m, 2H); ES-LCMS m/z 574.1 [M+H] ⁺ .

Step 2: tert -Butyl 1-(4- aminophenyl )-3- cyclopropyl -5-(2- fluoro- 4- iodo -anilino)-6,8-dimethyl-pyrido[4,3- d ]pyrimidine-2,4,7-trione

5-(4-Aminoanilino)-3-cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-pyrido[2,3-] in THF (0.3 mL) d ] A mixture of pyrimidine-2,4,7-trione (100 mg, 163.95 μmol, 1 eq) and NaOMe/MeOH (163.95 μmol, 11 μL, 30.0%, 1 eq) was degassed and purified with N ₂ . After purging for 1 hour, the mixture was stirred at 25° C. for 1 hour under N ₂ atmosphere. The reaction mixture was quenched by adding AcOH (0.1 mL), then diluted with H ₂ O (20 mL) and extracted using EtOAc (30 mL x 3). 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 analyzed by preparative TLC (DCM/MeOH = 10/1, TLC: DCM). /MeOH=10/1, R _f = 0.80) to give tert -butyl 1-(4-aminophenyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6. ,8-Dimethyl-pyrido[4,3- d ]pyrimidine-2,4,7-trione (90 mg, 67.50 μmol, 41.2% yield, 43.0% purity) was obtained as a yellow solid. ES-LCMS m/z 574.0 [M+H] ⁺ .

Step 3: tert -Butyl 3- cyclopropyl -5-(2- fluoro -4- iodo - Anilino )-6,8-dimethyl-1-[4-(methylsulfamoylamino)phenyl]pyrido[4,3- d ]Pyrimidine-2,4,7-trione

1-(4-aminophenyl)-3-cyclopropyl-5-(2-fluoro-4-iodo-anilino)-6,8-dimethyl-pyrido[4,3-] in DCM (1 mL) d ]pyrimidine-2,4,7-trione (43.00 mg, 75.00 μmol, 1 eq) in a solution of TEA (15.18 mg, 149.99 μmol, 20.88 μL, 2 eq) and N -methylsulfamoyl chloride (9.72 mg). , 75.00 μmol, 1 eq) was added. The mixture was stirred at 0°C for 1 hour. The reaction mixture was quenched by addition of H ₂ O (20 mL) and extracted using EtOAc (30 mL x 3). 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 subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um; mobile phase: [water (FA)-ACN]; B%: 56%-76%, 12 min) and then lyophilized to give tert -butyl 3-cyclopropyl-5-(2-fluoro- 4-iodo-anilino)-6,8-dimethyl-1-[4-(methylsulfamoylamino)phenyl]pyrido[4,3- d ]pyrimidine-2,4,7-trione (8.77 mg, 13.16 μmol, 17.6% yield, 100.0% purity) was obtained as a white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 11.23-10.93 (m, 1H), 10.00-9.81 (m, 1H), 7.90-7.70 (m, 1H), 7.59-7.52 (m, 1H), 7.50-7.44 (m, 1H), 7.29-7.23 (m, 2H), 7.22-7.15 (m, 2H), 6.99-6.82 (m, 1H), 3.12-3.01 (m, 3H), 2.63-2.57 (m) , 1H), 2.47-2.44 (m, 3H), 1.22 (s, 3H), 0.99-0.91 (m, 2H), 0.72-0.57 (m, 2H); ES-LCMS m/z 667.0 [M+H] ⁺ .

4-(3- cyclopropyl -5-((2- fluoro -4- Iodophenyl )Amino)-6,8-dimethyl-2,4,7- Trioxo -3,4,6,7-tetrahydropyrido[4,3- d ]Pyrimidine-1(2 H )-Day)- N -methyl-1 H -Synthesis of indole-1-sulfonamide (I-28)

step 1: 5 -((One H -Indole-4-yl)amino)-3- cyclopropyl -1-(2- fluoro -4- Iodophenyl )-6,8-dimethylpyrido[2,3- d ]Pyrimidine-2,4,7(1 H , 3H ,8 H ) - trion

[3-Cyclopropyl-1-(2-fluoro-4-iodo-phenyl)-6,8-dimethyl-2,4,7-trioxo-pyrido[2,3-] in DMF (5 mL) d ]pyrimidin-5-yl] trifluoromethanesulfonate (1.2 g, 1.92 mmol, 98.6%, 1 eq) in a solution of 1 H -indol-4-amine (254.15 mg, 1.92 mmol, 1 eq) and 2,6-lutidine (618.14 mg, 5.77 mmol, 671.90 μL, 3 eq) was added. The mixture was stirred at 60° C. under a microwave for 8 hours. The reaction mixture was diluted with H ₂ O (30 mL), extracted using EtOAc (50 mL x 3) and washed with brine (40 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue which was purified by flash silica gel chromatography (PE/EtOAc = from 100/1 to 1/1, TLC: PE/EtOAc = 1/1, R _f = 0.2) to obtain 5-((1 H -indol-4-yl)amino)-3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6, 8-dimethylpyrido[2,3- d ]pyrimidine-2,4,7( 1H , 3H , 8H )-trione (900 mg, 1.40 mmol, 72.9% yield, 93.0% purity) Obtained as a green solid. ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 11.22 (s, 1H), 10.46 (s, 1H), _7.95 (d, J = 9.4 Hz, 1H), 7.74 (d, J = 8.2 Hz, 1H) ), 7.58-7.49 (m, 1H), 7.34 (d, J = 2.5 Hz, 1H), 7.13 (d, J = 8.0 Hz, 1H), 7.03-7.00 (m, 2H), 6.40 (d, J = 7.4 Hz, 1H), 2.75 (s, 3H), 2.67 (dd, J = 3.3, 6.8 Hz, 1H), 1.45 (s, 3H), 0.99 (d, J = 6.7 Hz, 2H), 0.67 (m, 2H); ES-LCMS m/z 598.0 [M+H] ⁺ .

step 2: 3 - cyclopropyl -5-((2- fluoro -4- Iodophenyl )Amino)-1-(1 H -Indole-4-yl)-6,8-dimethylpyrido[4,3- d ]Pyrimidine-2,4,7(1 H ,3 H ,6 H ) - trion

5-((1 H -indol-4-yl)amino)-3-cyclopropyl-1-(2-fluoro-4-iodophenyl)-6,8-dimethylpyrido[ 2,3- d ]pyrimidine-2,4,7( 1H , 3H , 8H )-trione (500 mg, 701.82 μmol, 83.9%, 1 eq) was added to a solution of NaOMe/MeOH (701.82 μmol, 0.1 mL, 30.0%, 1 eq) was added. The mixture was stirred at 25°C for 0.5 hours under N ₂ atmosphere. CH ₃ COOH was added to the residue until pH = 7 and extracted using EtOAc (40 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 analyzed by preparative TLC (PE/EtOAc = 1/1, TLC: PE/EtOAc = 1/1, R _f = 0.3) to obtain 3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-1-( 1H -indol-4-yl)-6,8-dimethylpyrido[ 4,3- d ]pyrimidine-2,4,7( 1H , 3H , 6H )-trione (330 mg, 469.55 μmol, 66.9% yield, 85.0% purity) was obtained as a grey-green solid. ^1H NMR (400 MHz, DMSO- d6 ) δ ppm 11.29 (s, 1H), 11.12 (s, 1H), 7.79 (dd, J = 1.7, 10.4 Hz _, 1H), 7.59-7.52 (m, 1H) , 7.45 (d, J = 8.1 Hz, 1H), 7.38 (t, J = 2.7 Hz, 1H), 7.12 (t, J = 7.8 Hz, 1H), 6.98-6.89 (m, 2H), 6.22 (s, 1H), 3.07 (s, 3H), 2.68-2.59 (m, 1H), 0.96 (s, 3H), 0.95-0.88 (m, 2H), 0.70 (d, J = 4.1 Hz, 2H); ES-LCMS m/z 598.0 [M+H] ⁺ .

Step 3: 4-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo-3,4,6, 7-Tetrahydropyrido[4,3- d ]Pyrimidine-1(2 H )-day)-1 H -indole-1-sulfonyl fluoride

3-Cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-1-(1 H -indol-4-yl)-6,8-dimethylpyrido[ DIEA (110.34 mg, 853.73 μmol) in a solution of 4,3- d ]pyrimidine-2,4,7(1 H , 3 H , 6 H )-trione (100 mg, 142.29 μmol, 85.0%, 1 eq) , 148.70 μL, 6 eq) was added. The suspension was degassed under vacuum and purged several times with N ₂ . The mixture was stirred at 25° C. under F ₂ O ₂ S (15 Psi) for 24 hours. The mixture was concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (HCl)-ACN]; B%: 70%-90% , 10 minutes) and lyophilized to obtain 4-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo. -3,4,6,7-tetrahydropyrido[4,3- d ]pyrimidin-1( 2H )-yl) -1H -indole-1-sulfonyl fluoride (20 mg, 29.44 μmol, 20.7% yield, non-measurable purity) was obtained as a white solid. ES-LCMS m/z 679.9 [M+H]+.

Step 4: 4-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo-3,4,6, 7-Tetrahydropyrido[4,3- d ]Pyrimidine-1(2 H )-Day)- N -methyl-1 H -indole-1-sulfonamide

4-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo-3,4 in DMF (2 mL) ,6,7-Tetrahydropyrido[4,3- d ]pyrimidin-1( 2H )-yl)-1H-indole-1-sulfonyl fluoride (20 mg, 29.44 μmol, not measurable, 1 eq. ) DIEA (49.46 mg, 382.67 μmol, 66.65 μL, 13 eq) and methanamine hydrochloride (39.75 mg, 588.73 μmol, 20 eq) were added to the solution. The mixture was stirred at 80°C for 12 hours. The reaction mixture was purified by preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (HCl)-ACN]; B%: 60%-80%, 10 min) and lyophilized. 4-(3-cyclopropyl-5-((2-fluoro-4-iodophenyl)amino)-6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetra Hydropyrido[4,3- d ]pyrimidin-1( 2H )-yl) -N -methyl- 1H -indole-1-sulfonamide (2.51 mg, 3.64 μmol, 12.4% yield, 100.0% purity) was obtained as a white solid. ^1H NMR (500 MHz, CDCl ₃ ) δ ppm 11.37 (s, 1H), 7.91 (d, J = 8.2 Hz, 1H), 7.56-7.51 (m, 2H), 7.47 (d, J = 8.7 Hz, 1H) ), 7.38 (t, J = 8.0 Hz, 1H), 7.19 (d, J = 7.6 Hz, 1H), 6.72 (t, J = 8.2 Hz, 1H), 6.46-6.31 (m, 1H), 4.88 (d , J = 5.0 Hz, 1H), 3.21 (s, 3H), 2.83-2.70 (m, 1H), 2.64 (d, J = 5.3 Hz, 3H), 1.27 (d, J = 6.1 Hz, 2H), 1.15 (s, 3H), 0.89-0.83 (m, 2H); ES-LCMS m/z 691.0 [M+H]+.

( S )- N' -(3-(6- cyclopropyl -4-((2- fluoro -4- Iodophenyl ) amino) -1,3-dimethyl-2,5-dioxo-1,2,5,6-tetrahydropyrido [2,3-d] pyridazin-8-yl) phenyl) -N-methylmethane Sulfonimideamide & ( R )- N' -(3-(6-cyclopropyl-4-((2-fluoro-4-iodophenyl)amino)-1,3-dimethyl-2,5-dioxo-1,2,5,6-tetra Hydropyrido[2,3-d]pyridazin-8-yl)phenyl)-N-methylmethanesulfonimideamide (I-119 & I- 120) synthesis

step 1: 2 -(3- Nitrophenyl )-2-oxo-acetic acid

To a solution of 2-oxo-2-phenyl-acetic acid (51 g, 339.70 mmol, 1 eq ) in H ₂ SO ₄ (80 mL) at 0° C. was added KNO ₃ (41.21 g, 407.64 mmol, 1.2 eq ). . The mixture was stirred at 25°C for 16 hours. The reaction mixture was poured into ice water (1.6 L) and extracted using EtOAc (400 mL x 3). The combined organic phases were washed with brine (300 mL), dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give 2-(3-nitrophenyl)-2-oxo-acetic acid (66 g, crude). was obtained as a yellow solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.70 (t, J = 1.8 Hz, 1H), 8.56 (td, J = 1.2, 8.2 Hz, 1H), 8.41-8.37 (m, 1H), 7.89 (t, J = 8.0 Hz, 1H).

Step 2: Ethyl 2-(3- Nitrophenyl )-2-oxo-acetate

To a solution of 2-(3-nitrophenyl)-2-oxo-acetic acid (66 g, 338.24 mmol, 1 eq ) in DMF (700 mL) was added K ₂ CO ₃ (121.54 g, 879.42 mmol, 2.6 eq ). . After stirring at 20°C for 2 hours, iodoethane (263.77 g, 1.69 mol, 135.26 mL, 5 eq ) was added dropwise. The mixture was stirred at 20°C for 16 hours. The reaction mixture was poured into water (800 mL) and extracted using EtOAc (500 mL x 3). The combined organic layers were washed with brine (500 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 (PE/EtOAc = 1/0 to 10/ 1, purified by TLC: PE/EtOAc = 5/1, R _f = 0.45) to give ethyl 2-(3-nitrophenyl)-2-oxo-acetate (35.5 g, 143.16 mmol, 42.3% yield, 90.0% purity). ) was obtained as a yellow oil. ^1H NMR (400MHz, CDCl ₃ ) δ ppm 8.90 (s, 1H), 8.54-8.47 (m, 1H), 8.40 (d, J = 7.8 Hz, 1H), 7.75 (t, J = 7.9 Hz, 1H) , 4.50 (q, J = 7.2 Hz, 2H), 1.46 (t, J = 7.1 Hz, 3H).

Step 3: Ethyl 3-[(2E)-2-[2- ethoxy -1-(3- Nitrophenyl )-2-oxo- ethylidene ] Hydrazino ]-3-oxo-propanoate

To a solution of ethyl 2-(3-nitrophenyl)-2-oxo-acetate (22.5 g, 90.73 mmol, 1 eq ) in EtOH (120 mL) was added H ₂ SO ₄ (2.29 g, 23.33 mmol, 1.24 mL, 0.25 eq) . ) and ethyl 3-hydrazino-3-oxo-propanoate (13.79 g, 94.36 mmol, 1.04 eq ) were added. The mixture was stirred at 90°C for 2 hours. The solvent was removed to give a residue, which was diluted with water (200 mL) and extracted using EtOAc (300 mL x 3). The combined organic phases were washed with brine (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give ethyl 3-[(2E)-2-[2-ethoxy-1-(3- Nitrophenyl)-2-oxo-ethylidene]hydrazino]-3-oxo-propanoate (30 g, crude, E/Z mixture) was obtained as a yellow oil. ES-LCMS m/z 352.2 [M+H] ⁺ .

Step 4: Ethyl 4- hydroxy -3-(3- Nitrophenyl )-6-oxo-1H- Pyridazine -5- carboxylate

Ethyl 3-[(2E)-2-[2-ethoxy-1-(3-nitrophenyl)-2-oxo-ethylidene]hydrazino]-3-oxo-propanoate in DMF (300 mL) To a solution of (30 g, 85.39 mmol, E/Z mixture, 1 eq ) was added K ₂ CO ₃ (6.49 g, 46.97 mmol, 0.55 eq ). The mixture was stirred at 80°C for 3 hours. The mixture was cooled to ambient temperature and poured into 3N HCl (600 mL). The precipitated solid was collected by filtration to obtain a residue, to which MeOH (50 mL) was added and stirred at 15°C for 2 hours. The slurry was filtered and the cake was rinsed with MeOH (3 x 3 mL). The solid was collected and dried in vacuo to yield ethyl 4-hydroxy-3-(3-nitrophenyl)-6-oxo-1H-pyridazine-5-carboxylate (8.3 g, 24.47 mmol, 28.7% yield, 90.0% Purity) was obtained as a yellow solid. ^1H NMR (400MHz, DMSO- d6 ) δ ppm 13.23 (s, 1H), 8.58-8.53 (m, 1H), 8.30 (td, J = 1.2, _7.2 Hz, 1H), 8.15 (d, J = 8.1) Hz, 1H), 7.76 (t, J = 8.1 Hz, 1H), 4.32 (q, J = 7.1 Hz, 2H), 1.29 (t, J = 7.1 Hz, 3H); ES-LCMS m/z 306.1 [M+H] ⁺ .

Step 5: Ethyl 4- chloro -3-(3- Nitrophenyl )-6-oxo-1 H - Pyridazine -5- carboxylate

To a solution of ethyl 4-hydroxy-3-(3-nitrophenyl)-6-oxo-1 H -pyridazine-5-carboxylate (2.5 g, 7.37 mmol, 1 eq) in DCM (30 mL) (COCl ) ₂ (935.60 mg, 7.37 mmol, 645.24 μL, 1 eq) was added. The mixture was stirred at 35°C for 3 hours. The reaction mixture was quenched by adding water (30 mL) and extracted using DCM (40 mL x 3). 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 to which MeOH (5 mL) was added and incubated at 25° C. for 2 h. It was stirred. The slurry was filtered and the cake was rinsed with PE (2 x 3 mL). The solid was collected and dried in vacuo to obtain ethyl 4-chloro-3-(3-nitrophenyl)-6-oxo-1H-pyridazine-5-carboxylate (1.9 g, 5.69 mmol, 77.2% yield, 97.0% purity). ) was obtained as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 11.67 (s, 1H), 8.48 (s, 1H), 8.38 (d, J = 8.1 Hz, 1H), 7.92 (d, J = 7.8 Hz, 1H), 7.74-7.66 (m, 1H), 4.51 (q, J = 7.1 Hz, 2H), 1.43 (t, J = 7.1 Hz, 3H); ES-LCMS m/z 324.0, 326.0 [M+H] ⁺ .

Step 6: Ethyl 5- chloro -2- cyclopropyl -6-(3- Nitrophenyl )-3-oxo- Pyridazine -4- carboxylate

Ethyl 4-chloro-3-(3-nitrophenyl)-6-oxo-1H-pyridazine-5-carboxylate (1.4 g, 4.20 mmol, 1 eq) in 1,4-dioxane (20 mL), cyclo Propylboronic acid (720.75 mg, 8.39 mmol, 2 eq), 2-(2-pyridyl)pyridine (1.64 g, 10.49 mmol, 2.5 eq), Na ₂ CO ₃ (533.60 mg, 5.03 mmol, 1.2 eq) and Cu A mixture of (OAc) ₂ (914.43 mg, 5.03 mmol, 1.2 eq) was degassed, purged three times with N ₂ and the mixture was stirred at 70° C. for 24 hours under N ₂ atmosphere. The reaction mixture was diluted with water (100 mL) and extracted using DCM (60 mL x 3). The combined organic phases were washed with brine (50 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 (PE/EtOAc = 1/0 to 3). /1, TLC: PE/EtOAc = 3/1, R _f = 0.43) to give ethyl 5-chloro-2-cyclopropyl-6-(3-nitrophenyl)-3-oxo-pyridazine-4- The carboxylate (400 mg, 989.69 μmol, 23.6% yield, 90.0% purity) was obtained as a yellow gum. ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 8.42 (d, J = 1.8 Hz, 1H), 8.35 (td, J = 1.1, 8.3 Hz, 1H), 7.87 (d, J = 7.6 Hz, 1H), 7.70-7.65 (m, 1H), 4.51 (q, J = 7.1 Hz, 2H), 4.22 (t, J = 4.0, 7.6 Hz, 1H), 1.43 (t, J = 7.2 Hz, 3H), 1.22-1.16 (m, 2H), 1.13-1.06 (m, 2H); ES-LCMS m/z 364.0, 366.0 [M+H] ⁺ .

Step 7: Ethyl 2- cyclopropyl -5-( Methylamino )-6-(3- Nitrophenyl )-3-oxo- Pyridazine -4-carboxylate

of ethyl 5-chloro-2-cyclopropyl-6-(3-nitrophenyl)-3-oxo-pyridazine-4-carboxylate (400 mg, 989.69 μmol, 1 eq) in DCM (3 mL) at 0°C. MeNH ₂ (1.02 g, 9.90 mmol, 30.0% purity, 10 eq) was added to the solution. The mixture was stirred at 20°C for 2 hours. The solvent was removed to obtain ethyl 2-cyclopropyl-5-(methylamino)-6-(3-nitrophenyl)-3-oxo-pyridazine-4-carboxylate (300 mg, 669.74 μmol, 67.7% yield, 80.0% Purity) was obtained as a yellow solid. ^1H NMR (500 MHz, CDCl ₃ ) δ ppm 8.33-8.30 (m, 2H), 7.79 (d, J = 7.6 Hz, 1H), 7.70-7.66 (m, 1H), 4.43 (q, J = 7.1 Hz) , 2H), 4.02-3.95 (m, 1H), 2.69 (d, J = 5.3 Hz, 3H), 1.44 (t, J = 7.1 Hz, 2H), 1.46-1.42 (m, 1H), 1.03-0.94 ( m, 4H); ES-LCMS m/z 359.1 [M+H] ⁺ .

step 8:2 - cyclopropyl -5-( Methylamino )-6-(3- Nitrophenyl ) Pyridazine -3-on

of ethyl 2-cyclopropyl-5-(methylamino)-6-(3-nitrophenyl)-3-oxo-pyridazine-4-carboxylate (300 mg, 669.74 μmol, 1 eq) in DMSO (5 mL) LiCl (283.93 mg, 6.70 mmol, 10 eq) was added to the solution. The mixture was stirred at 150°C for 8 hours. The reaction mixture was diluted with water (30 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 (PE/EtOAc = 1/0 to 0). /1, TLC: PE/EtOAc = 0/1, R _f = 0.15) to give 2-cyclopropyl-5-(methylamino)-6-(3-nitrophenyl)pyridazin-3-one (200 mg, 628.74 μmol, 93.9% yield, 90.0% purity) was obtained as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 8.38-8.27 (m, 2H), 7.83 (d, J = 7.6 Hz, 1H), 7.73-7.66 (m, 1H), 5.92 (s, 1H), 4.03 (t, J = 3.9, 7.5 Hz, 1H), 2.83 (d, J = 3.4 Hz, 3H), 1.09-0.93 (m, 4H); ES-LCMS m/z 287.1 [M+H] ⁺ .

step 9:6 - cyclopropyl -4- hydroxy -1,3-dimethyl-8- (3-nitrophenyl)pyrido[2,3-d]pyridazine -2,5-dione

To a solution of 2-cyclopropyl-5-(methylamino)-6-(3-nitrophenyl)pyridazin-3-one (200 mg, 628.74 μmol, 1 eq) in Ac ₂ O (3 mL) was added 2-methyl. Propanedioic acid (222.74 mg, 1.89 mmol, 152.56 μL, 3 eq) was added. The mixture was stirred at 110°C for 2 hours. The solvent was removed to give a residue, which was purified by preparative TLC (PE/EtOAC = 3/1, TLC: PE/EtOAC = 3/1, R _f = 0.24) to give 6-cyclopropyl-4-hydroxy -1,3-dimethyl-8-(3-nitrophenyl)pyrido[2,3-d]pyridazine-2,5-dione (200 mg, 434.38 μmol, 69.1% yield, 80.0% purity) was obtained as a white solid. was obtained. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 12.80 (s, 1H), 8.37 (td, J = 2.2, 7.0 Hz, 1H), 8.33 (d, J = 1.5 Hz, 1H), 7.72-7.68 (m , 2H), 4.16-4.09 (m, 1H), 3.06 (s, 3H), 2.16 (s, 3H), 1.20-1.15 (m, 2H), 1.15-1.09 (m, 2H); ES-LCMS m/z 369.0 [M+H] ⁺ .

Step 10: [6- cyclopropyl -1,3-dimethyl-8-(3- Nitrophenyl )-2,5- dioxo - Pyrido[2,3-d]pyridazine -4-day] 4- Methylbenzenesulfonate

6-cyclopropyl-4-hydroxy-1,3-dimethyl-8-(3-nitrophenyl)pyrido[2,3-d]pyridazine-2,5-dione (200 mg) in DCM (8 mL) , 434.38 μmol, 1 eq), DIEA (561.40 mg, 4.34 mmol, 756.61 μL, 10 eq) and 4-methylbenzenesulfonyl chloride (414.06 mg, 2.17 mmol, 5 eq) were added. The mixture was stirred at 50°C for 12 hours. The solvent was removed to obtain a residue, which was purified by preparative TLC (PE/EtOAC = 3/1, TLC: PE/EtOAC = 3/1, R _f = 0.24) to give [6-cyclopropyl-1,3 -Dimethyl-8-(3-nitrophenyl)-2,5-dioxo-pyrido[2,3-d]pyridazin-4-yl] 4-methylbenzenesulfonate (200 mg, 371.27 μmol, 85.5% Yield, 97.0% purity) was obtained as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.38-8.30 (m, 2H), 7.98 (d, J = 8.3 Hz, 2H), 7.77-7.63 (m, 2H), 7.41 (d, J = 8.0 Hz) , 2H), 4.22-4.06 (m, 1H), 3.08 (s, 3H), 2.49 (s, 3H), 1.85 (s, 3H), 1.12-1.01 (m, 4H); ES-LCMS m/z 523.1 [M+H] ⁺ .

step 11:6 - cyclopropyl -4-(2- fluoro -4- iodo - Anilino )-1,3-dimethyl-8- (3-nitrophenyl)pyrido[2,3-d]pyridazine -2,5-dione

To a solution of 2-fluoro-4-iodo-aniline (263.99 mg, 1.11 mmol, 3 eq) in THF (3 mL) was added NaH (89.10 mg, 2.23 mmol, 60.0% purity, 6 eq), After stirring at 0°C for 30 min, [6-cyclopropyl-1,3-dimethyl-8-(3-nitrophenyl)-2,5-dioxo-pyrido[2,3-d]pyri minced-4-yl] 4-methylbenzenesulfonate (200 mg, 371.27 μmol, 1 eq) was added. The reaction temperature was slowly warmed to 25°C and stirred at 25°C for 1 hour. The reaction mixture was quenched by adding water (20 mL) and extracted using EtOAc (30 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 analyzed by preparative TLC (PE/EtOAC = 1/1, TLC: PE/ EtOAC = 1/1, R _f = 0.35) to give 6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-8-(3-nitrophenyl) Pyrido[2,3-d]pyridazine-2,5-dione (100 mg, 156.64 μmol, 42.2% yield, 92.0% purity) was obtained as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ ppm 10.78 (s, 1H), 8.44-8.27 (m, 2H), 7.75-7.65 (m, 2H), 7.46 (d, J = 1.8, 10.0 Hz, 1H) , 7.37 (d, J = 8.5 Hz, 1H), 6.56 (t, J = 8.5 Hz, 1H), 4.10-4.01 (m, 1H), 3.07 (s, 3H), 1.82 (s, 3H), 1.19- 1.07 (m, 4H); ES-LCMS m/z 588.0 [M+H] ⁺ .

step 12:8 -(3- Aminophenyl )-6- cyclopropyl -4-(2- fluoro -4- iodo - Anilino )-1,3-dimethyl-pyrido[2,3-d]pyridazine-2,5-dione

6-Cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-8- in EtOH (1.5 mL), H ₂ O (1.5 mL) and THF (1.5 mL). (3-nitrophenyl)pyrido[2,3-d]pyridazine-2,5-dione (100 mg, 156.64 μmol, 1 eq) was added to a solution of NH ₄ Cl (83.79 mg, 1.57 mmol, 10 eq) and Fe (43.74 mg, 783.19 μmol, 5 eq) was added. The mixture was stirred at 90°C for 3 hours. The reaction mixture was filtered to remove insoluble material. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was subjected to preparative HPLC (column: Boston Prime C18 150*30 mm*5 um; mobile phase: [water (NH ₃ H ₂ O + NH ₄ HCO ₃ )-ACN]; B%: 64%-94%, 10 minutes) and then lyophilized to produce 8-(3-aminophenyl)-6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-pyrido. [2,3-d]pyridazine-2,5-dione (55.23 mg, 99.09 μmol, 63.3% yield, 100.0% purity) was obtained as a yellow solid. ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 7.55 (d, J = 1.8, 10.3 Hz, 1H), 7.45 (d, J = 8.5 Hz, 1H), 7.21 (t, J = 7.7 Hz, 1H), 6.82 (d, J = 1.5, 8.0 Hz, 1H), 6.79-6.76 (m, 1H), 6.71 (d, J = 7.7 Hz, 1H), 6.64 (t, J = 8.6 Hz, 1H), 4.04 (t , J = 3.9, 7.5 Hz, 1H), 3.13 (s, 3H), 1.76 (s, 3H), 1.17-1.11 (m, 2H), 1.07-0.99 (m, 2H); ES-LCMS m/z 558.0 [M+H] ⁺ .

Step 13: Methanesulfinyl chloride

SO _{2 Cl 2} ( 20.27 g, 170.38 mmol, 12.36 mL, 3 eq) was added. The mixture was stirred at 20°C for 12 hours. The reaction mixture was concentrated to give crude methanesulfinyl chloride (8.9 g, crude) as a light yellow oil, which was used in the next step without further purification. ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 3.39 (s, 3H).

Step 14: N- [3-[6- cyclopropyl -4-(2- fluoro -4- iodo - Anilino )-1,3-dimethyl-2,5-dioxo-pyrido[2,3-d]pyridazin-8-yl]phenyl] Methanesulfinamide

8-(3-aminophenyl)-6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-pyrido[2,3-] in DCM (30 mL) d]pyridazine-2,5-dione (500 mg, 897.09 μmol, 100%, 1 eq) mixed with DIEA (907.77 mg, 7.02 mmol, 1.22 mL, 7.83 eq) and methanesulfinyl chloride (176.82 mg, 1.79 mg). mmol, 2 eq) was added. The mixture was stirred at 20°C for 1 hour. TLC (PE/EtOAc = 1/1, R _f = 0.07) showed that the starting material was completely consumed and one new spot was formed. The reaction mixture was concentrated to give a residue, which was purified by flash silica gel chromatography (PE/EtOAc = 1/0 to 0/1, TLC: PE/EtOAc = 1/1, R _f = 0.07) to give N -[3-[6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-2,5-dioxo-pyrido[2,3-d]pyrido Minced-8-yl]phenyl]methanesulfinamide (350 mg, 553.72 μmol, 61.7% yield, 98.0% purity) was obtained as a yellow solid. ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 10.85 (br s, 1H), 7.48-7.33 (m, 3H), 7.23-7.02 (m, 3H), 6.54 (t, J = 7.8 Hz, 1H), 6.33 (br s, 1H), 4.05 (br s, 1H), 3.11 (s, 3H), 2.87 (s, 3H), 1.82 (s, 3H), 1.25-1.05 (m, 4H); ES-LCMS m/z 620.2 [M+H] ⁺ .

Step 15: 6-Cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-8-[3-[[methyl-(methylamino)-oxo-λ6-sulfa Nylidene] amino] phenyl] pyrido [2,3-d] pyridazine-2,5-dione

N-[3-[6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-2,5 in MeCN (45 mL) at -20°C under N ₂ -dioxo-pyrido[2,3-d]pyridazin-8-yl]phenyl]methanesulfinamide (150 mg, 237.31 μmol, 98%, 1 eq) and MeNH ₂ (0.35 g, 3.72 mmol, 33% , 15.67 eq), t-BuOK (1 M, 1.42 mL, 6 eq) was added, followed by NCS (126.75 mg, 949.23 umol, 4 eq). The mixture was stirred at 20°C for 3 hours. TLC (PE/EtOAc = 0/1, R _f = 0.28) showed that the starting material was completely consumed and one new spot was formed. 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. The residue was purified by flash silica gel chromatography (PE/EtOAc = 1/0 to 0/1, TLC: PE/EtOAc = 0/1, R _f = 0.28) to give 6-cyclopropyl-4-(2- Fluoro-4-iodo-anilino)-1,3-dimethyl-8-[3-[[methyl-(methylamino)-oxo-λ6-sulfanylidene]amino]phenyl]pyrido[2,3 -d]pyridazine-2,5-dione (80 mg, 109.79 μmol, 46.2% yield, 89.0% purity) was obtained as a white solid. ES-LCMS m/z 649.2 [M+H] ⁺ .

Step 16: ( S )- N' -(3-(6-cyclopropyl-4-((2-fluoro-4-iodophenyl)amino)-1,3-dimethyl-2,5-dioxo-1,2,5,6-tetra Hydropyrido[2,3-d]pyridazin-8-yl)phenyl)-N-methylmethanesulfonimideamide & ( R )- N' -(3-(6-cyclopropyl-4-((2-fluoro-4-iodophenyl)amino)-1,3-dimethyl-2,5-dioxo-1,2,5,6-tetra Hydropyrido[2,3-d]pyridazin-8-yl)phenyl)-N-methylmethanesulfonimideamide

6-Cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-8-[3-[[methyl-(methylamino)-oxo-λ6-sulfanylidene] Amino]phenyl]pyrido[2,3-d]pyridazine-2,5-dione (80 mg, 109.79 μmol, 89%, 1 eq) was isolated by SFC (AD_IPA_DEA_5_40_4ML_4MIN_5CM). The desired fractions were concentrated to obtain a residue, which was then lyophilized to yield peak 1: ( S ) -N '-(3-(6-cyclopropyl-4-((2-fluoro-4-iodophenyl)amino )-1,3-dimethyl-2,5-dioxo-1,2,5,6-tetrahydropyrido[2,3-d]pyridazin-8-yl)phenyl)-N-methylmethanesulfone Midamide (20.27 mg, 30.91 μmol, 28.1% yield, 98.9% purity) (ee=100%) was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.58-7.52 (m, 1H), 7.45 (d, J = 8.2 Hz, 1H), 7.34 (t, J = 7.9 Hz, 1H), 7.19 (d, J = 7.8 Hz, 1H), 7.15 (s, 1H), 7.07 (d, J = 7.5 Hz, 1H), 6.65 (t, J = 8.5 Hz, 1H), 4.05 (td, J = 3.6, 7.4 Hz, 1H), 3.12 (d, J = 7.0 Hz, 6H), 2.72 (s, 3H), 1.77 (s, 3H), 1.14 (d, J = 2.7 Hz, 2H), 1.03 (d, J = 5.2 Hz, 2H); ES-LCMS m/z 649.2 [M+H] ⁺ . Peak 2: ( R ) -N '-(3-(6-cyclopropyl-4-((2-fluoro-4-iodophenyl)amino)-1,3-dimethyl-2,5-dioxo- 1,2,5,6-Tetrahydropyrido [2,3-d] pyridazin-8-yl) phenyl) - N -methylmethanesulfonimideamide (20.68 mg, 31.67 μmol, 28.8% yield, 99.3% Purity) (ee=100%) was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 7.55 (dd, J = 1.8, 10.3 Hz, 1H), 7.45 (d, J = 8.5 Hz, 1H), 7.34 (t, J = 7.9 Hz, 1H) , 7.19 (d, J = 7.9 Hz, 1H), 7.15 (s, 1H), 7.07 (d, J = 6.7 Hz, 1H), 6.65 (t, J = 8.6 Hz, 1H), 4.05 (tt, J = 3.9, 7.5 Hz, 1H), 3.18-3.06 (m, 6H), 2.72 (s, 3H), 1.77 (s, 3H), 1.20-1.09 (m, 2H), 1.07-0.98 (m, 2H); ES-LCMS m/z 649.2 [M+H] ⁺ .

1-[3-[6- cyclopropyl -4-(2- fluoro -4- iodo - Anilino )-1,3-dimethyl-2,5- dioxo -pyrido[2,3-d]pyridazin-8-yl]phenyl]-3- methyl -Synthesis of guanidine (I-122)

Step 1: Ethyl N -[[3-[6- cyclopropyl -4-(2- fluoro -4- iodo - Anilino )-1,3-dimethyl-2,5-dioxo-pyrido[2,3-d]pyridazin-8-yl]phenyl]carbamothioyl]carbamate

8-(3-aminophenyl)-6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-pyrido[2,3-] in DCM (10 mL) d]pyridazine-2,5-dione (300 mg, 538.25 μmol, 100.0%, 1 eq) and ethyl N -(thioxomethylene)carbamate (155.31 mg, 1.18 mmol, 2.2 eq) at 15°C. It was stirred for 12 hours. To the mixture was added PE, filtered and concentrated under reduced pressure to give ethyl N -[[3-[6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl. -2,5-dioxo-pyrido[2,3-d]pyridazin-8-yl]phenyl]carbamothioyl]carbamate (330 mg, 460.12 μmol, 85.5% yield, 96.0% purity) was obtained as a yellow solid. was obtained, which was used in the next step without further purification. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 11.58 (s, 1H), 10.83 (br s, 1H), 8.12 (s, 1H), 7.86 (s, 1H), 7.65 (d, J = 7.9 Hz, 1H), 7.51 (t, J = 7.9 Hz, 1H), 7.45 (d, J = 9.9 Hz, 1H), 7.35 (d, J = 8.4 Hz, 1H), 7.30-7.27 (m, 1H), 6.54 (t, J = 8.5 Hz, 1H), 4.32 ( q, J = 7.0 Hz, 2H), 4.06 (tt, J = 3.9, 7.4 Hz, 1H), 3.17 (s, 3H), 1.82 (s, 3H), 1.38 (t, J = 7.0 Hz, 3H), 1.19-1.14 (m, 2H), 1.10-1.04 (m, 2H); ES-LCMS m/z 689.2 [M+H] ⁺ .

Step 2: Ethyl N -[( Z )- N -[3-[6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-2,5-dioxo-pyrido[2,3-d]pyrido minced-8-yl]phenyl]-- N '-methyl-carbamimidoyl]carbamate

Ethyl N -[[3-[6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-2,5-di in DCM (20 mL) at 0°C. Oxo-pyrido[2,3-d]pyridazin-8-yl]phenyl]carbamothioyl]carbamate (270 mg, 376.47 μmol, 96.0% purity, 1 eq), DIEA (316.25 mg, 2.45 mmol, 426.22 EDCI (288.67 mg, 1.51 mmol, 4 eq) was added to a solution of (μL, 6.5 eq) and methanamine;hydrochloride (101.67 mg, 1.51 mmol, 4 eq). The mixture was stirred at 15°C for 12 hours. TLC (DCM/MeOH = 10/1, R _f = 0.50) showed complete consumption of starting material. The mixture was diluted with water (30 mL) and extracted using DCM (20 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 (DCM/MeOH = from 100/1 to 50/1, TLC: DCM/MeOH = 10/1, R _f = 0.50) to obtain ethyl N -[(Z)- N -[3-[6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1 ,3-dimethyl-2,5-dioxo-pyrido[2,3-d]pyridazin-8-yl]phenyl] -N '-methyl-carbamimidoyl]carbamate (260 mg, 371.71 μmol, 98.7 % yield, 98.0% purity) was obtained as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 10.81 (s, 1H), 7.54-7.18 (m, 6H), 6.53 (t, J = 8.5 Hz, 1H), 4.19 (q, J = 7.2 Hz, 2H), 4.08-4.00 (m, 1H), 3.10 (s, 3H), 2.97 (d, J = 4.6 Hz, 3H), 1.81 (s, 3H), 1.37-1.29 (m, 3H), 1.18-1.11 (m, 2H), 1.11-1.04 (m, 2H) ); ES-LCMS m/z 686.1 [M+H] ⁺ .

step 3: 1 -[3-[6- cyclopropyl -4-(2- fluoro -4- iodo - Anilino )-1,3-dimethyl-2,5-dioxo-pyrido[2,3-d]pyridazin-8-yl]phenyl]-3- methyl -Guanidine

Ethyl N -[( Z )- N -[3-[6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-2 in EtOH (4 mL), 5-dioxo-pyrido[2,3-d]pyridazin-8-yl]phenyl] -N' -methyl-carbamimidoyl]carbamate (100 mg, 142.96 μmol, 98.0% purity, 1 eq) KOH (2 M, 1.14 mL, 16 eq) was added to the solution. The mixture was stirred at 40°C for 12 hours. The mixture was concentrated under reduced pressure to obtain a residue, which was subjected to preparative HPLC (column: Boston Green ODS 150*30 mm*5 um; mobile phase: [water (FA)-ACN]; B%: 26%-46% , 12 minutes) and then lyophilized to produce 1-[3-[6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-2,5- Dioxo-pyrido[2,3-d]pyridazin-8-yl]phenyl]-3-methyl-guanidine (15.28 mg, 23.17 μmol, 16.2% yield, 100.0% purity, FA) was obtained as a yellow solid. . ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.64-7.59 (m, 1H), 7.56 (d, J = 10.4 Hz, 1H), 7.47 (d, J = 8.2 Hz, 2H), 7.44-7.34 (m, 2H), 6.66 (t, J = 8.5 Hz) , 1H), 4.13-4.01 (m, 1H), 3.10 (s, 3H), 2.94 (s, 3H), 1.77 (s, 3H), 1.16-1.09 (m, 2H), 1.08-1.02 (m, 2H) ); ES-LCMS m/z 614.1 [M+H] ⁺ .

( S )- N '-(3-(6- cyclopropyl -4-((2- fluoro -4- Iodophenyl )Amino)-1,3-dimethyl-2,5-dioxo-1,2,5,6-tetrahydropyrido[2,3- d ]pyridazine-8-yl)phenyl)- N , N -Synthesis of dimethylmethanesulfonimideamide (I-123)

Step 1: N -[3-[6- cyclopropyl -4-(2- fluoro -4- iodo - Anilino )-1,3-dimethyl-2,5-dioxo-pyrido[2,3- d ]pyridazin-8-yl]phenyl]methanesulfinamide

8-(3-aminophenyl)-6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-pyrido[2,3-] in DCM (2 mL) d ]pyridazine-2,5-dione (100 mg, 179.42 μmol, 100.0%, 1 eq), methanesulfinyl chloride (26.52 mg, 269.13 μmol, not measurable, 1.5 eq) and DIEA (69.57 mg, 538.25 μmol, The mixture (93.75 μL, 3 eq) was degassed and purged three times with N ₂ and then the mixture was stirred at 0° C. for 10 min under N ₂ atmosphere. The reaction mixture was washed with saturated NaHCO ₃ (30 mL) solution and extracted using DCM (30 mL x 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give N -[3-[6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3. -Dimethyl-2,5-dioxo-pyrido[2,3- d ]pyridazin-8-yl]phenyl]methanesulfinamide (110 mg, 163.37 μmol, 91.1% yield, 92.0% purity) as a yellow solid. Obtained, which was used directly in the next step without further purification. ES-LCMS m/z 620.0 [M+H] ⁺ .

Step 2: ( S )- N '-(3-(6-cyclopropyl-4-((2-fluoro-4-iodophenyl)amino)-1,3-dimethyl-2,5-dioxo-1,2,5,6- Tetrahydropyrido[2,3- d ]pyridazine-8-yl)phenyl)- N , N -Dimethylmethanesulfonimideamide

N -[3-[6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-2,5-dioxo-pyrido[ in MeCN (30 mL) 2,3- d ]pyridazin-8-yl]phenyl]methanesulfinamide (110 mg, 163.37 μmol, 92.0%, 1 eq) in a solution of N -methylmethanamine (2 M, 816.86 μL, 10 eq) and t-BuOK (1 M, 1.31 mL, 8 eq) was added followed by NCS (141.80 mg, 1.06 mmol, 6.5 eq). The mixture was stirred at 40° C. for 12 hours under N ₂ atmosphere. The reaction mixture was diluted with H ₂ O (30 mL) and extracted using EtOAc (30 mL x 3). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was purified by preparative TLC (TLC: PE/EtOAc = 2/3, R _f = 0.2). The desired fractions were concentrated under reduced pressure to obtain a residue, 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%: 40%-40%) to obtain peak 1 and peak 2. Peak 1 was concentrated under reduced pressure to obtain a residue, which was washed with MeCN (20 mL) and water (20 mL). Dissolved in and lyophilized to ( S ) -N '-(3-(6-cyclopropyl-4-((2-fluoro-4-iodophenyl)amino)-1,3-dimethyl-2,5- Dioxo-1,2,5,6-tetrahydropyrido[2,3- d ]pyridazin-8-yl)phenyl) -N , N -dimethylmethanesulfonimideamide (18.06 mg, 27.26 μmol, 16.7 % yield, 100.0% purity, SFC: R _t = 5.737, ee = 100.0%) was obtained as a white solid, ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 10.87 (s, 1H), 7.45 (dd, J = 1.8, 10.0 Hz, 1H), 7.35 (d, J = 9.7 Hz, 1H), 7.31 (d, J = 8.0 Hz, 1H), 7.21 (d, J = 8.9 Hz, 1H), 7.14-6.97 (m , 2H), 6.53 (t, J = 8.5 Hz, 1H), 4.05 (s, 1H), 3.12 (s, 3H), 2.99 (s, 3H), 2.91 (s, 6H), 1.81 (s, 3H) , 1.19-1.13 (m, 2H), 1.09-1.01 (m, 2H); ES-LCMS m/z 663.1 [M+H] ⁺ .

( R )- N '-(3-(6- cyclopropyl -4-((2- fluoro -4- Iodophenyl )Amino)-1,3-dimethyl-2,5-dioxo-1,2,5,6-tetrahydropyrido[2,3- d ]pyridazine-8-yl)phenyl)- N , N -Synthesis of dimethylmethanesulfonimideamide (I-124)

Step 1: ( R )- N '-(3-(6-cyclopropyl-4-((2-fluoro-4-iodophenyl)amino)-1,3-dimethyl-2,5-dioxo-1,2,5,6- Tetrahydropyrido[2,3- d ]pyridazine-8-yl)phenyl)- N , N -Dimethylmethanesulfonimideamide

N -[3-[6-cyclopropyl-4-(2-fluoro-4-iodo-anilino)-1,3-dimethyl-2,5-dioxo-pyrido[ in MeCN (30 mL) 2,3- d ]pyridazin-8-yl]phenyl]methanesulfinamide (110 mg, 163.37 μmol, 92.0%, 1 eq) in a solution of N -methylmethanamine (2 M, 816.86 μL, 10 eq) and t-BuOK (1 M, 1.31 mL, 8 eq) was added followed by NCS (141.80 mg, 1.06 mmol, 6.5 eq). The mixture was stirred at 40°C for 12 hours under N ₂ atmosphere. The reaction mixture was diluted with H ₂ O (30 mL) and extracted using EtOAc (30 mL x 3). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was purified by preparative TLC (TLC: PE/EtOAc = 2/3, R _f = 0.2). The desired fractions were concentrated under reduced pressure to obtain a residue, 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%: 40%-40%) to obtain peak 1 and peak 2. Peak 2 was concentrated under reduced pressure to obtain a residue, which was washed with MeCN (20 mL) and water (20 mL). Dissolve in and lyophilize to ( R ) -N '-(3-(6-cyclopropyl-4-((2-fluoro-4-iodophenyl)amino)-1,3-dimethyl-2,5- Dioxo-1,2,5,6-tetrahydropyrido[2,3- d ]pyridazin-8-yl)phenyl) -N , N -dimethylmethanesulfonimideamide (19.63 mg, 28.75 μmol, 17.6 % yield, 97.0% purity, SFC: R _t = 6.270, ee = 97.7%) was obtained as a white solid, ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 10.87 (s, 1H), 7.45 (dd, J =1.8, 10.0 Hz, 1H), 7.35 (d, J = 9.7 Hz, 1H), 7.31 (d, J = 8.0 Hz, 1H), 7.21 (d, J = 8.2 Hz, 1H), 7.14-6.98 (m , 2H), 6.53 (t, J = 8.5 Hz, 1H), 4.04 (d, J = 3.6 Hz, 1H), 3.12 (s, 3H), 2.99 (s, 3H), 2.91 (s, 6H), 1.81 (s, 3H), 1.18-1.13 (m, 2H), 1.08-1.02 (m, 2H); ES-LCMS m/z 663.1 [M+H] ⁺ .

6- cyclopropyl -4-[(2- fluoro -4- Iodophenyl )amino]-3- methyl -8- {3-[(methylsulfamoyl)amino]phenyl}pyrano Synthesis of [2,3-d]pyridazine-2,5-dione (I-129)

Step 1

n-BuLi (41.70 mL, 104.26 mmol, 6.00 eq) was added dropwise to a solution of DIPA (10.55 g, 104.26 mmol, 6.00 eq) in THF (20.00 mL) at -78°C under N ₂ atmosphere. The reaction mixture was stirred at -50°C for 0.5 hours. A solution of tert-butyl propionate (6.79 g, 52.13 mmol, 3.00 eq) in THF (20.00 mL) was added dropwise to the previous reaction mixture at -78°C and stirred for an additional 1 hour at -78°C. Then, ethyl 2-cyclopropyl-5-hydroxy-6-(3-nitrophenyl)-3-oxopyridazine-4-carboxylate (6.00 g, 17.38 mmol, 1.00 eq) in THF (120.00 mL) The solution was added dropwise at -78°C and the mixture was stirred at -50°C for an additional 2 hours. The reaction was quenched with saturated NH ₄ Cl (500 mL) and then extracted using EtOAc (2 x 200 mL). The combined organics were washed with brine (400 mL), dried over anhydrous Na ₂ SO ₄ and concentrated under vacuum to give tert-butyl 3-[2-cyclopropyl-5-hydroxy-6-(3-nitrophenyl)- 3-Oxopyridazin-4-yl]-2-methyl-3-oxopropanoate (6.00 g, crude) was obtained as a yellow solid.

Step 2

In a 100 mL round bottom flask, tert-butyl 3-[2-cyclopropyl-5-hydroxy-6-(3-nitrophenyl)-3-oxopyridazin-4-yl]-2 in TFA (50.00 mL) A solution of -methyl-3-oxopropanoate (6.00 g, 13.97 mmol, 1.00 eq) was stirred at 60°C for 3 hours. The reaction mixture was concentrated under vacuum to give the crude product, which was directly purified by flash chromatography (PE:EA=1:1) to give 6-cyclopropyl-3-methyl-8-(3-nitrophenyl)-3H. -Pyrano[2,3-d]pyridazine-2,4,5-trione (1.70 g, 34.27%) was obtained as a yellow solid.

Step 3

6-cyclopropyl-4-hydroxy-3-methyl-8-(3-nitrophenyl)pyrano[2,3-d]pyridazine-2,5-dione (1.70) in DCM (20.00 mL) at 0°C. g, 4.79 mmol, 1.00 eq) and TEA (1.33 mL, 9.57 mmol, 2.00 eq) was added dropwise to a solution of TsCl (1.09 g, 5.72 mmol, 1.20 eq) in DCM (20.00 mL). The mixture was stirred at 25°C for 16 hours. The reaction mixture was treated by adding 100.00 mL H ₂ O and extracted using EA (3 x 100.00 mL). The combined organic layers were washed with brine (150.00 mL) and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure to obtain the crude product, which was directly purified by flash chromatography (PE:EA=3:1) to give 6-cyclopropyl-3-methyl-8-(3-nitrophenyl) -2,5-dioxopyrano[2,3-d]pyridazin-4-yl 4-methylbenzenesulfonate (1.70 g, 51.55%) was obtained as a yellow solid.

Step 4

LiHMDS (10.01 mL, 10.01 mmol, 3.00 eq) was added dropwise to a solution of 2-fluoro-4-iodoaniline (2.85 g, 12.01 mmol, 3.60 eq) in THF (30.00 mL) at -78°C. It was stirred at °C for 0.5 hours. Then, 6-cyclopropyl-3-methyl-8-(3-nitrophenyl)-2,5-dioxopyrano[2,3-d]pyridazine-4- in THF (30.00 mL) at -78°C. A solution of 4-methylbenzenesulfonate (1.70 g, 3.34 mmol, 1.00 eq) was added dropwise to the reaction and stirred at -78°C for 0.5 h. The reaction mixture was treated by adding saturated NH ₄ Cl (150.00 mL) and extracted using EA (3 x 100.00 mL). The combined organic layers were washed with brine (150.00 mL) and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure to obtain the crude product, which was directly purified by flash chromatography (PE:EA=5:1) to give 6-cyclopropyl-4-[(2-fluoro-4-io Dophenyl)amino]-3-methyl-8-(3-nitrophenyl)pyrano[2,3-d]pyridazine-2,5-dione (1.41 g, 73.58%) was obtained as a yellow solid.

Step 5

6-cyclopropyl-4-[(2-fluoro-4-iodophenyl)amino]-3-methyl-8-(3-nitrophenyl)pyrano[2,3] in AcOH (20.00 mL) at 25°C. Fe (1.03 g, 18.5 mmol, 10.00 eq) was added to a solution of -d]pyridazine-2,5-dione (1.06 g, 1.85 mmol, 1.00 eq), and stirred at 50°C for 12 hours. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in 50.00 mL saturated NaHCO ₃ and extracted using EA (3 x 50.00 mL). The combined organic layers were washed with brine (50.00 mL) and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure to obtain 6-cyclopropyl-4-[(2-fluoro-4-iodophenyl)amino]-3-methyl-8-(3-nitrophenyl)pyrano[2, 3-d]pyridazine-2,5-dione (0.78 g, 73.58%) was obtained as a yellow solid, which was used directly in the next step without further purification.

Step 6

8-(3-aminophenyl)-6-cyclopropyl-4-[(2-fluoro-4-iodophenyl)amino]-3-methylpyrano[2,3] in DMF (1.50 mL) at 0°C. -d]pyridazine-2,5-dione (0.24 g, 0.44 mmol, 1.00 eq) and pyridine (0.11 g, 1.32 mmol, 3.00 eq) in a solution of N-methylsulfamoyl chloride in CH ₃ CN (1.50 mL). (0.09 g, 0.66 mmol, 1.50 eq) was added dropwise. The resulting mixture was stirred at 0°C for 0.5 hours. The reaction mixture was treated by adding 20.00 mL H ₂ O and extracted using EA (3 x 30.00 mL). The combined organic layers were washed with brine (50.00 mL) and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by preparative-HPLC ( _column : _Welch , mobile phase B: ACN; flow rate: 90 mL/min; gradient: 90% B to 90% B in 10 min, 90% B; wavelength: 254 nm; RT1 (min): 7; number of runs: 5) Purified to obtain 6-cyclopropyl-4-[(2-fluoro-4-iodophenyl)amino]-3-methyl-8-{3-[(methylsulfamoyl)amino]phenyl}pyrano[2,3 -d]pyridazine-2,5-dione (62 mg, 33.09%) was obtained as a yellow solid. LCMS: (ES, m/z) [M+H] ⁺ : 638.1. ¹ H NMR: (400 MHz, DMSO- d ₆ ): δ 7.72 - 7.71 (m, 1H), 7.63 - 7.60 (m, 1H), 7.57 - 7.50 (m, 2H), 7.46 - 7.42 (m, 1H) , 7.35 - 7.32 (m, 1H), 6.92 (t, J =8.4 Hz, 1H), 4.20 - 4.16 (m, 1H), 2.62 (s, 3H), 1.65 (s, 3H), 1.25 - 1.23 (m , 2H), 1.11 - 1.06 (m, 2H).

6- cyclopropyl -3- fluoro -4-[(2- fluoro -4- Iodophenyl )Amino]-1- methyl Synthesis of -8-{3-[(methylsulfamoyl)amino]phenyl}pyrido[2,3-d]pyridazine-2,5-dione (I-128)

Step 1

Stirring ethyl 2-cyclopropyl-5-hydroxy-6-(3-nitrophenyl)-3-oxopyridazine-4-carboxylate (3.5 g, 1.45 mmol, 1.0 eq) in DCM (30 mL) at room temperature. (COCl) ₂ (12.9 g, 14.48 mmol, 10 eq) and DMF (0.7 mL, 1.30 mmol, 0.9 eq) were added dropwise to the solution. The resulting mixture was stirred at 40°C for 1 hour. Solvent and excess oxalyl dichloride were removed under reduced pressure. The residue was layered with DCM (50 mL) and H ₂ O (30 mL), and the water layer was extracted using DCM (2*30 mL). The combined organic layers were washed with brine (50 mL) and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure to afford ethyl 5-chloro-2-cyclopropyl-6-(3-nitrophenyl)-3-oxopyridazine-4-carboxylate (5.3 g, crude) as a brown oil. Obtained. ES-LCMS m/z 364.0 [M+H] ⁺

Step 2

Stirring of ethyl 5-chloro-2-cyclopropyl-6-(3-nitrophenyl)-3-oxopyridazine-4-carboxylate (4 g, 11.00 mmol, 1.0 eq) in DCM (40 mL) at 0°C. CH ₃ NH ₂ /THF (45 mL, 90.00 mmol, 8.18 eq) was added dropwise to the solution. The resulting mixture was stirred at room temperature for an additional 4 hours. The solvent was removed under reduced pressure. The residue was diluted in H ₂ O (100 mL) and extracted using DCM (3 x 100 mL). The combined organic layers were washed with brine (100 mL) and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure to obtain ethyl 2-cyclopropyl-5-(methylamino)-6-(3-nitrophenyl)-3-oxopyridazine-4-carboxylate (5.06 g, crude). Obtained as a tan solid. ES-LCMS m/z 359.2 [M+H] ⁺

Step 3

Ethyl 2-cyclopropyl-5-(methylamino)-6-(3-nitrophenyl)-3-oxopyridazine-4-carboxylate (3.6 g, 10.05 mmol, 1.0 eq) in ACN (36 mL) at room temperature. Ethyl 3-chloro-3-oxopropanoate (3.6 mL) was added in portions to the stirred solution. The resulting mixture was stirred at 80°C for an additional 2 hours. The reaction mixture was cooled and quenched by pouring into water/ice (150 mL). The resulting mixture was extracted using EtOAc (3 x 150 mL). The combined organic layers were washed with brine (150 mL) and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE/THF (5:1) to give ethyl 2-cyclopropyl-5-(3-ethoxy-N-methyl-3-oxopropanamido)-6- (3-Nitrophenyl)-3-oxopyridazine-4-carboxylate (4.42 g, 93%) was obtained as a brown solid. ES-LCMS m/z 473.2 [M+H] ⁺

Step 4

Ethyl 2-cyclopropyl-5-(3-ethoxy-N-methyl-3-oxopropanamido)-6-(3-nitrophenyl)-3-oxo in MeOH (45 mL) at 0°C under air. Sodium methoxide (1.0 g, 18.63 mmol, 2.0 eq) was added in portions to a stirred solution of pyridazine-4-carboxylate (4.4 g, 9.31 mmol, 1.0 eq). The resulting mixture was stirred at 0°C for an additional 1 hour. The resulting mixture was concentrated, acidified to pH 4 with 1N HCl (aq) and extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (50 mL) and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure to obtain ethyl 6-cyclopropyl-1-methyl-8-(3-nitrophenyl)-2,4,5-trioxo-3H-pyrido[2,3-d]pyrido. Minced-3-carboxylate (3.7 g, 92%) was obtained as a brown solid. ES-LCMS m/z 427.2 [M+H] ⁺

Step 5

Ethyl 6-cyclopropyl-1-methyl-8-(3-nitrophenyl)-2,4,5-trioxo-3H-pyrido[2,3-d) in ACN (35 mL) at 0°C under air. ]Selectfluor (3.3 g, 9.23 mmol, 1.5 eq) was added in portions to a stirred solution of pyridazine-3-carboxylate acetate (3.1 g, 6.16 mmol, 1.0 eq) and sodium acetate (1.0 g, 12.31 mmol, 2.0 eq). did. The resulting mixture was stirred at 0°C for an additional 3 hours. The reaction was quenched by adding water/ice (5 mL) at 0°C and then extracted using EtOAc (3 x 10 mL). The combined organic layers were washed with brine (10 mL) and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure to obtain ethyl 6-cyclopropyl-3-fluoro-1-methyl-8-(3-nitrophenyl)-2,4,5-trioxopyrido[2,3-d ]Pyridazine-3-carboxylate (2.4 g, 88%) was obtained as a yellow solid. ES-LCMS m/z 445.1 [M+H] ⁺ .

Step 6

Ethyl 6-cyclopropyl-3-fluoro-1-methyl-8-(3-nitrophenyl)-2,4,5-trioxopyrido[2,3-) in THF (25 mL) at room temperature under air. To a stirred solution of d]pyridazine-3-carboxylate (2.4 g, 5.45 mmol, 1.0 eq), H ₂ SO ₄ (24 mL) was added dropwise. The resulting mixture was stirred at 90°C for 3 hours. THF was then removed and the residue was diluted with water (10 mL) and extracted using DCM (3 x 20 mL). The combined organic layers were washed with brine (20 mL) and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE/THF (5:1) to give 6-cyclopropyl-3-fluoro-4-hydroxy-1-methyl-8-(3-nitrophenyl)pyri. Do[2,3-d]pyridazine-2,5-dione (1.1 g, 54%) was obtained as a yellow solid. ES-LCMS m/z 373.1 [M+H] ⁺ .

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 13.12 (s, 1H), 8.44 - 8.41 (m, 1H), 8.40 - 8.34 (m, 1H), 8.04 - 7.98 (m, 1H), 7.87 - 7.79 (m, 1H), 3.78-3.70 (m, 1H), 2.93 (s, 3H), 1.10 - 1.07 (m, 2H), 1.07 - 1.04 (m, 2H).

Step 7

6-cyclopropyl-3-fluoro-4-hydroxy-1-methyl-8-(3-nitrophenyl)pyrido[2,3-d]pyridazine-2 in DCM (10 mL) at 0°C, To a stirred solution of 5-dione (1.1 g, 2.96 mmol, 1.0 eq) and pyridine (0.7 g, 8.87 mmol, 3.0 eq) was added dropwise Tf ₂ O (1.7 g, 5.91 mmol, 2.0 eq) in DCM (2 mL). did. The resulting mixture was stirred at room temperature for 0.5 hours. The mixture was diluted with water (10 mL) and extracted using EtOAc (3 x 20 mL). The combined organic layers were washed with brine (20 mL) and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE/THF (5:1) to give 6-cyclopropyl-3-fluoro-1-methyl-8-(3-nitrophenyl)-2,5-di. Oxopyrido[2,3-d]pyridazin-4-yl trifluoromethanesulfonate (460 mg, 30%) was obtained as a yellow solid. ES-LCMS m/z 505.1 [M+H] ⁺ .

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.47 (s, 1H), 8.39 - 8.32 (m, 1H), 8.10 - 8.03 (m, 1H), 7.88 - 7.79 (m, 1H), 4.10-4.0 (m, 1H), 2.97 (s, 3H), 1.49 - 1.22 (m, 2H), 0.94 - 0.76 (m, 2H).

Step 8

To a stirred solution of 2-fluoro-4-iodoaniline (648 mg, 2.74 mmol, 3.0 eq) in THF (2 mL) at 0°C under nitrogen atmosphere was added LDA (2M in THF) (1.82 ml, 3.65 mmol, 4.0 eq) was added dropwise. The resulting mixture was stirred at 0°C for 30 minutes under nitrogen atmosphere. To the above mixture was added 6-cyclopropyl-3-fluoro-1-methyl-8-(3-nitrophenyl)-2,5-dioxopyrido[2,3-d] in THF (3 mL) at 0°C. Pyridazin-4-yl trifluoromethanesulfonate (460 mg, 0.91 mmol, 1.0 eq) was added dropwise. The resulting mixture was stirred at room temperature for an additional 30 minutes. The reaction was monitored by LCMS. The reaction was quenched by addition of water (20 mL) at 0°C and extracted using EtOAc (3 x 20 mL). The combined organic layers were washed with brine (30 mL) and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE/EA (2:1) to give 6-cyclopropyl-3-fluoro-4-[(2-fluoro-4-iodophenyl)amino]- 1-Methyl-8-(3-nitrophenyl)pyrido[2,3-d]pyridazine-2,5-dione (210 mg, 39%) was obtained as a white solid. ES-LCMS m/z 592.0 [M+H] ⁺ .

Step 9

6-cyclopropyl-3-fluoro-4-[(2-fluoro-4-iodophenyl)amino]-1-methyl-8-(3-nitrophenyl)pyrido in AcOH (5 mL) at room temperature. Fe (226 mg, 4.06 mmol, 10 eq) was added in portions to a stirred solution of [2,3-d]pyridazine-2,5-dione (240 mg, 0.40 mmol, 1.0 eq). The resulting mixture was stirred at 60°C for 30 minutes under a nitrogen atmosphere. The reaction was monitored by LCMS. The resulting mixture was cooled to room temperature, diluted with water (20 mL), neutralized to pH 7 using saturated NaHCO ₃ (aq), and extracted using EtOAc (3 x 20 mL). The combined organic layers were washed with brine (30 mL) and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE/EA (1:1) to give 8-(3-aminophenyl)-6-cyclopropyl-3-fluoro-4-[(2-fluoro- 4-Iodophenyl)amino]-1-methylpyrido[2,3-d]pyridazine-2,5-dione (150 mg, 66%) was obtained as a white solid. ES-LCMS m/z 562.0 [M+H] ⁺ .

Step 10

8-(3-aminophenyl)-6-cyclopropyl-3-fluoro-4-[(2-fluoro-4-iodophenyl)amino]-1 in DCM (1.5 mL) at 0°C under nitrogen atmosphere. -Methylpyrido[2,3-d]pyridazine-2,5-dione (150 mg, 0.27 mmol, 1.0 eq) and Et ₃ N (81 mg, 0.80 mmol, 3.0 eq) were added to a stirred solution in DCM (0.5 N-methylsulfamoyl chloride (69 mg, 0.53 mmol, 2.0 eq) in mL) was added dropwise. The resulting mixture was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. The reaction was quenched by addition of water (20 mL) at 0°C and extracted using EtOAc (3 x 20 mL). The combined organic layers were washed with brine (20 mL) and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure. The crude product (200 mg) was incubated under the following conditions (complete non-WJ SC hardware 50 mm x 40 cm, 7 μm, 40 - 60% 18 min, NH ₃ H ₂ O + NH ₄ HCO ₃ /CAN, 80 mL/min Flash) for preparative purification by HPLC to obtain 6-cyclopropyl-3-fluoro-4-[(2-fluoro-4-iodophenyl)amino]-1-methyl-8-{3-[(methyl Sulfamoyl)amino]phenyl}pyrido[2,3-d]pyridazine-2,5-dione (68.4 mg, 39%) was obtained as a white solid. ES-LCMS m/z 655.0 [M+H] ⁺ .

^OneH NMR (400 MHz, DMSO-d6) δ 10.92 (d,J = 2.6 Hz, 1H), 9.87 (s, 1H), 7.76 - 7.68 (m, 1H), 7.59 - 7.52 (m, 1H), 7.46 - 7.35 (m, 2H), 7.30 - 7.22 (m, 2H), 7.20 - 7.13 (m, 1H), 7.06 - 6.96 (m, 1H), 4.03 - 3.93 (m, 1H), 2.93 (s, 3H), 2.47 (d,J = 4.8 Hz, 3H), 1.06 - 0.99 (m, 4H).

6- cyclopropyl -4-[(2- fluoro -4- Iodophenyl )amino]-8- {3-[(methylsulfamoyl)amino]phenyl} Synthesis of pyridazino[4,5-e][1,3]oxazine-2,5-dione (I-127)

Step 1

Methyl 2-cyclopropyl-5-hydroxy-6-(3-nitrophenyl)-3-oxopyridazine-4-carboxylate (5 g, 15.093 mmol, 1 eq) in 1,4-dioxane (40 mL) ) and NH ₃ *H ₂ O (10 mL, 256.805 mmol, 17.12 eq) was stirred at 80°C for 2 hours. The resulting mixture was concentrated under reduced pressure and purified by trituration with ethyl ether (30 mL). This gave 2-cyclopropyl-5-hydroxy-6-(3-nitrophenyl)-3-oxopyridazine-4-carboxamide (4.6 g, 96.37%) as a light brown solid.

Step 2

2-Cyclopropyl-5-hydroxy-6-(3-nitrophenyl)-3-oxopyridazine-4-carboxamide (4.6 g, 14.544 mmol, 1 eq) and CDI (3.54 g, 21.816 mmol) in DMF , 1.5 eq) was stirred at room temperature overnight. The precipitated solid was collected by filtration and washed with MeOH (3 x 20 mL). As a result, 6-cyclopropyl-4-hydroxy-8-(3-nitrophenyl)pyridazino[4,5-e][1,3]oxazine-2,5-dione (4.2 g, 84.37%) was obtained. Obtained as a yellow solid.

Step 3

6-cyclopropyl-4-hydroxy-8-(3-nitrophenyl)pyridazino[4,5-e][1,3]oxazine-2,5-dione in DCE (4.2 g, 12.271 mmol, A mixture of 1 eq) and PCl ₅ (5.11 g, 24.542 mmol, 2 eq) was stirred at 80°C for 2 hours under a nitrogen atmosphere. The solvent was removed under reduced pressure to obtain 4-chloro-6-cyclopropyl-8-(3-nitrophenyl)pyridazino[4,5-e][1,3]oxazine-2,5-dione (6 g, Crude) was obtained as a yellow solid. The crude product was used directly in the next step without further purification.

Step 4

2-fluoro-4-iodoaniline (5.91 g, 24.951 mmol, 1.5 eq) and 4-chloro-6-cyclopropyl-8-(3-nitrophenyl)pyridazino[4,5-e] in acetonitrile. ][1,3]A mixture of oxazine-2,5-dione (6 g, 16.634 mmol, 1 eq) was stirred at room temperature under a nitrogen atmosphere for 2 hours. The precipitated solid was collected by filtration and washed with ACN (3 x 10 mL). The collected solid was purified by trituration with MeOH (20 mL). Thereby, 6-cyclopropyl-4-[(2-fluoro-4-iodophenyl)amino]-8-(3-nitrophenyl)pyridazino[4,5-e][1,3]oxazine- 2,5-dione (3.8 g, 55.15%) was obtained as a yellow solid.

Step 5

6-cyclopropyl-4-[(2-fluoro-4-iodophenyl)amino]-8-(3-nitrophenyl)pyridazino[4,5-e][1, 3] A mixture of oxazine-2,5-dione (250 mg, 0.445 mmol, 1 eq) and SnCl ₂ (2.54 g, 13.230 mmol, 29.73 eq) was stirred at room temperature overnight. The resulting solution was diluted with EtOAc (50 mL) and washed with 3 x 20 mL of water. The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. Thereby, 8-(3-aminophenyl)-6-cyclopropyl-4-[(2-fluoro-4-iodophenyl)amino]pyridazino[4,5-e][1,3]oxazine- 2,5-dione (280 mg, crude) was obtained as a yellow-green solid.

Step 6

8-(3-aminophenyl)-6-cyclopropyl-4-[(2-fluoro-4-iodophenyl)amino]pyridazino[4,5-e] in DMF (2.8 mL) at 0°C. To a stirred solution of [1,3]oxazine-2,5-dione (280 mg, 0.527 mmol, 1 eq) and pyridine (125.06 mg, 1.581 mmol, 3 eq) N-methylsulfamoyl in ACN (1 mL) Chloride (102.42 mg, 0.790 mmol, 1.5 eq) was added dropwise. The resulting mixture was stirred at room temperature for 30 minutes and then diluted with EtOAc (2*20 mL). The combined organic phases were washed with 3 x 20 mL of water and concentrated under vacuum. The residue was subjected to preparative HPLC (column: Welch Ultimate 6-cyclopropyl-4-[(2-fluoro-4-iodophenyl)amino]-8 purified from 30% B to 65% B in minutes, 65% B in 9 minutes; wavelength: 254 nm.) -{3-[(methylsulfamoyl)amino]phenyl}pyridazino[4,5-e][1,3]oxazine-2,5-dione (66.9 mg, 20.33%) was obtained as an off-white solid. . LCMS:(ES, m/z)[M+H] ⁺ : 625.1. ¹ H NMR: (300 MHz, DMSO- _d6 ) δ 12.02 (br, s, 1H), 9.93 (br, s, 1H), 8.29-8.18 (m, 1H), 7.92-7.82 (m, 1H), 7.76 -7.67 (m, 1H), 7.58 (s, 1H), 7.50-7.42 (m, 2H), 7.41-7.28 (m, 2H), 4.24-4.13 (m, 1H), 2.50 (s, 3H), 1.19 -1.05 (m, 4H).

Although a number of embodiments of the invention have been described, it will be clear that the basic example can 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 represented by example.

Claims (26)

A compound of formula (I') or a pharmaceutically acceptable salt thereof:

In the above equation,
Ring A is is selected from;
each independently represents a single bond or a double bond;
each R ¹ is independently H, halogen, -CN, or optionally substituted C _1-6 aliphatic;
X is C, CH, or N;
Y is CR ⁶ , C(O), or N;
L' is a covalent bond, -O-, -C(R ⁹ ) ₂ -, or -NR ⁸ -;
Each of R ² , R ⁵ , and R ⁸ is independently H or an optionally substituted C _1-6 aliphatic;
Each of R ³ , R ⁴ , R ⁶ , and R ⁹ is independently H, halogen, or optionally substituted C _1-6 aliphatic;
Ring B is a 3-8 membered monocyclic carbocyclic ring, a 3-8 membered monocyclic heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S, 5-11 a 5-11 membered bicyclic carbocyclic ring having 1-5 heteroatoms independently selected from a 1-membered bicyclic carbocyclic ring, N, O, or S, a phenyl ring, N, O, or S a 5-6 membered monocyclic heteroaromatic ring having 1-3 heteroatoms independently selected from, an 8-11 membered bicyclic aromatic ring, and 1-5 members independently selected from N, O, or S. an optionally substituted ring selected from 8-11 membered bicyclic heteroaromatic rings having heteroatoms;
L is a covalent bond, or 1, 2 or 3 methylene units of the chain are independently and optionally -O-, -S-, -N(R)-, -N ⁺ (R) ₂ -, -CH(R )-, -CH(OR)-, -CH(SR)-, -CH(N(R) ₂ )-, -C(=NR)-, -C(=N-OR)-, -C(O )-, -S(O) ₂ -, -(CH ₂ -CH ₂ -O) _1-10 -, -S(O)-, -N(R)-C(O)-, -N(R) -S(O) ₂ -, -C(O)-N(R)-, -S(O) ₂ -N(R)-, -P(O)(OR)-, or -Cy- C _1-10 2 is a straight or branched hydrocarbon chain;
Each -Cy- is independently a 3-7 membered carbocyclic ring, a 3-7 membered heterocyclic ring having 1-3 heteroatoms independently selected from N, O, or S, a phenyl ring, and N is an optionally substituted ring selected from a 5-6 membered heteroaromatic ring having 1-3 heteroatoms independently selected from , O, or S;
R ⁷ is R, -CN, -S(O) ₂ -N(R) ₂ , -NR-S(O) ₂ -R, -P(O)(OR)-N(R) ₂ , -C( =NR)-N(R) ₂ , -C(=N-OR)-N(R) ₂ , -C(O)-R, -S(O)-R, -N(R) ₂ , -OR , -SR, or ego;
Each R is independently H, -CN, or optionally substituted C _1-6 aliphatic;
n is 0, 1, 2, 3, 4, or 5. The method of claim 1, wherein ring A is Phosphorus compounds. The method of claim 1, wherein ring A is Phosphorus compounds. The method of claim 1, wherein ring A is Phosphorus compounds. The method of claim 1, wherein ring A is Phosphorus compounds. The compound according to any one of claims 1 to 5, wherein each R ¹ is independently halogen. The compound according to any one of claims 1 to 6, wherein R ² is H. The compound of any one of claims 1 to 7, wherein R ³ is optionally substituted C _1-6 alkyl. The compound of any one of claims 1 to 8, wherein R ⁴ is optionally substituted C _1-6 alkyl. The compound of any one of claims 1 to 9, wherein R ⁵ is an optionally substituted 3, 4, 5, or 6 membered saturated or unsaturated carbocyclyl. 11. The compound of any one of claims 1-10, wherein R ⁶ is optionally substituted C _1-6 alkyl. 12. The compound of any one of claims 1-11, wherein ring B is an optionally substituted phenyl ring. 12. The compound of any one of claims 1-11, wherein ring B is an optionally substituted 5-6 membered heteroaromatic ring having 1-3 heteroatoms independently selected from N, O, or S. 12. The compound of any one of claims 1-11, wherein ring B is an optionally substituted 8-10 membered bicyclic heteroaromatic ring having 1-5 heteroatoms independently selected from N, O, or S. 15. The method of any one of claims 1 to 14, wherein L is 1, 2 or 3 methylene units of the chain independently and optionally -O-, -N(R)-, -CH(R)-, -C( C _1- replaced by =NR)-, -C(=N-OR)-, -C(O)-, -S(O) ₂ -, -P(O)(OR)-, or -Cy- ₁₀ Compounds with divalent straight or branched hydrocarbon chains. 16. The method of any one of claims 1 to 15, wherein -Cy- is an optionally substituted 3, 4, 5, 6, or 7 membered group having 1, 2, or 3 heteroatoms independently selected from N, O, or S. A compound that is a heterocyclic ring of. The method according to any one of claims 1 to 16, wherein R ⁷ is -CN, -S(O) ₂ -N(R) ₂ , -NR-S(O) ₂ -R, -P(O)(OR)- N(R) ₂ , -C(=NR)-N(R) ₂ , -C(=N-OR)-N(R) ₂ , -OR, or Phosphorus compounds. The compound of any one of claims 1 to 17, wherein n is 2. The compound according to any one of claims 1 to 18, wherein the compound is a compound of formula (II) , (III) , or (IV) : or a pharmaceutically acceptable salt thereof:
. The method of any one of claims 1 to 19, wherein the compound has the following formulas (II-a) to (II-o) , (III-a) to (III-e) , and (IV-a) to (IV- A compound selected from e) , or a pharmaceutically acceptable salt thereof:


. 21. The compound according to any one of claims 1 to 20, wherein the compound is selected from the compounds in Table 1. A pharmaceutical composition comprising the compound of any one of claims 1 to 21, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle. A method of treating a disorder mediated by MEK, comprising:
A method comprising administering to a subject in need thereof a therapeutically effective amount of the compound of any one of claims 1 to 21, or the pharmaceutical composition of claim 22, to treat the disorder. 24. The method of claim 23, wherein the disorder is cancer. As a method of treating cancer in a patient,
A method comprising administering the compound of any one of claims 1 to 21, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 22 to a patient. As a method of inhibiting MEK activity,
A method comprising contacting MEK or a KSR-MEK complex or a RAF-MEK complex with an effective amount of the compound of any one of claims 1 to 21, or the pharmaceutical composition of claim 22 to inhibit MEK activity.